FlowAccess TM V3 USER MANUAL. Windows software for Skalar SAN plus systems. Version and up

Transcription

1 FlowAccess TM V3 Windows software for Skalar SAN plus systems Version and up USER MANUAL

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3 FlowAccess TM V3 Windows software for Skalar SAN plus systems Version and up User Manual Publication No: H.US Issue date: May 2013 Copyright Skalar Analytical B.V. P.O Box DE Breda The Netherlands Tel: +31 (0) Fax: +31 (0) info@skalar.com Internet:

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5 Welcome to the FlowAccess V3 Windows Software This manual guides you through the complete definition of the system, to help you quickly gain the necessary knowledge and confidence to use the FlowAccess V3 Windows Software to its optimum efficiency. Chapter 1 Introduction to the FlowAccess V3 software Chapter 2 System set-up guide Chapter 3 Active System window Chapter 4 Tables Chapter 5 Methods Chapter 6 Groups Chapter 7 Preparing for Analysis Chapter 8 The Analysis Run Chapter 9 DataProcessing guide In the Appendix some practical problems are outlined with their solution. Please read this manual carefully before the software is set into operation. If despite the information given in this manual, any problems or questions should arise, please do not hesitate to call your local Skalar representative or the Skalar head office at the address given below. Skalar Analytical B.V. P.O. Box DE BREDA The Netherlands Tel: Fax: info@skalar.com Internet:

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7 TABLE OF CONTENTS 1 INTRODUCTION TO THE FLOWACCESS V3 SOFTWARE General Information Short overview Software installation System requirements PC requirements Installation instructions Software Update Introduction Installation of an update Updating System Database Updating run files When a PC is renamed after installation of FlowAccess V Reinstalling a database after renaming the PC Communication settings The Main Window Login Restore a pre-defined system setup Path settings Main window items The File menu Update Configuration settings Log Off The Settings menu Interface Settings User Management Preferences CFR part Database Management Search in Runs The Help menu SYSTEM SETUP Introduction Setup of the system Adding the hardware Connecting the hardware Single channel connection Status settings Edit a system Delete a system ACTIVE SYSTEM

8 3.1 Introduction The Menu bar The File menu The Table menu The Analysis menu The Methods menu The Instruments menu The Other Settings menu Toolbar Status settings Control Changing the status Changing the settings in a certain status: Changing the sampler settings Tray settings Needle height settings Pre-dilution Utility Drift Cup details TABLES Introduction Sample table The Table Entry screen Table wizard Creating a table with the Table Wizard Creating a table by means of the Table menu Creating a table with the Table tool Creating a table in the Table Entry screen Creating a table for 2 or more needles Creating a table with automatic pre-dilutions Creating a table with spikes Importing tables Exporting tables Edit table Remove Table METHODS Methods menu Methods tool Create New Method Peak Detection parameters Peak Detection type Calibration Results QC & CLP concentration Post Dilution Load an existing method Edit a Method Custom method Remove a Method GROUPS Create New Group Remove Group Edit Group Group Details Working Standards settings Pre-dilution settings

9 6.3.4 Spiking Standard settings Post-Dilution settings CLP Action Fail status Group Information PREPARING FOR ANALYSIS Selection Baseline checking Digital matrix detector setup Time programming Time program Entry Enable a Time Program Set secondary time program Preparing a run from an external macro ANALYSIS RUN Starting the analysis run Viewing incoming results Viewing the calibration curves Viewing the run in Realtime Format Fields Result Format Custom chemistry Direct formula Beer Calculation Editing during the run Editing in Results screen Editing the calibration curve Table editing Method editing Overlapping analyses Stopping the analysis during the run Restoring a crashed run file End of the run Storing the run files manually DATA PROCESSING Introduction Importing runs Exporting runs Analysis Run info Comments Run groups FlowAccessV3Post Data Evaluation Real time screen Peak information and editing Multiple peak detection Single peak detection Peak picking parameters Break information Peak labels More Views Results screen Options menu Detailed results Cumulative results

10 9.4.4 Calibration Statistics Used formulas in statistics Edit Table & Method Table editing Method editing Reports APPENDIX

11 FlowAccess V3 CHAPTER 1: INTRODUCTION Preface: This chapter includes general information, software installation and requirements and security options.

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13 Chapter 1 Introduction to the FlowAccess V3 software INTRODUCTION TO THE FLOWACCESS V3 SOFTWARE 1.1 General Information The FlowAccess V3 Software is the latest multitasking data acquisition and instrument control software package for the Skalar SAN plus system operating under Windows XP, Windows Vista or Windows 7. The features of the basic program include: Automatic preparation of working standards (not for SA1000 or SA1100 sampler) Automatic zeroing of baseline Automatic start-up and shut-down of the system Automatic pre and post dilutions (not for SA1000 or SA1100 sampler) Automatic baseline and sensitivity drift correction Supports multi sampler systems Password security for various entry level access Full data protection Multiple view screens and real time data evaluation. This basic program can be extended with the following options that are embedded (if installed) in the program: - 21 CFR part 11 for the guarantee of reliable and validated results - Bar code reading for automatic identification of the samples - FlowExport TM for generating an RS232 output to an external PC These options are explained in separate manuals. Two other programs are available that (if installed) can be reached from within the FlowAccess V3 program: - QAccess TM (for quality control of the results) - FlowReport TM (for extended reports of the results) 1.2 Short overview FlowAccess distinguishes itself by the use of a simplified interaction between operator and system. Complete layout and connections of the analyzer are displayed on screen and fully controlled by the software, thus eliminating operator s errors. This layout is easily implemented by the use of the drag and drop technique. The hardware setup is usually performed once at initial setup and only recalled if the hardware is changed. In the Analysis screen the methods are defined. When the system and the methods are defined, groups of chemistries can be defined for analysis. For easy sample layout Sample type buttons with color codes are used. To enable quick setup of the table a Table Wizard can be used. By means of the Table Wizard a standard layout can be made for the calibration standards and the drift/wash intervals for the unknowns. By adding the total

14 FlowAccess V3 1-2 number of unknowns the table can be created. The combined sample table will be displayed. The table can be printed. The data collection takes place in the background while the analysis run is proceeding. This allows the operator to prepare new analysis runs or to do a variety of data handling tasks. The incoming signals can be displayed during the run in the real time data screen; before the run actually starts the operator can check the stability of the baseline. During the run, all the analysis peaks can be viewed, each channel singly or all channels simultaneously or 16 paged. During analysis the table can be opened for editing (e.g. excluding a wrong standard). The program can handle bubblegating analysis. Over range samples can be automatically post-diluted and analyzed (if a diluter is present on the sampler). A calibration curve is generated from a series of standard solutions increasing in concentration. The sample peaks are compared to this curve. Calibration curves can be calculated with a first or second order curve, through origin or not or reverse logarithm and can be corrected for baseline and sensitivity drift. The results of the compared sample peaks are automatically multiplied by the (possible) dilution factor and divided by the sample weight factor. The raw data are stored in the database and can be recalled at any time for further calculation. Recalculation with altered parameters is always possible as the method parameters can be edited from the data processing status. Also peaks can be reviewed and edited. The system performs automatic start-up and shutdown during or outside working hours. This increases the analyzer capacity and decreases reagent consumption. The software can be secured against unwanted editing by defining different levels of access to the functions of the software. Note: It is highly recommended not to open any other program during an analysis run. Warning: Whenever the program crashes by means of an error, the Communication Module will keep running on the Start bar. Close this program. FlowAccess V3 can only be restarted when this program is closed. 1.3 Software installation The Skalar software package FlowAccess V3 TM will be preinstalled on the computer supplied by Skalar. Important note: If other programs beside FlowAccess V3 are being installed Skalar does not guarantee the proper functioning of FlowAccess V3. If other computers than computers supplied by Skalar are used, Skalar will not guarantee the proper working of FlowAccess V System requirements If you install FlowAccess V3 as an update for an existing system, always first contact Skalar for investigation of your system, as update on your system may be required. Updates may be required for samplers (EPROM/motors), rinsing valves (220 volt, plugs), interfaces (EEPROMS) and detectors (plugs).

15 Chapter 1 Introduction to the FlowAccess V3 software PC requirements Minimum requirements: Processor RAM memory Video memory Hard disk drive Available space COM port USB port Parallel port DVD/ ROM drive Display monitor at least 2 GHz 4 GB 128 MB at least 250 GB 100 MB at least 1 x RS232 (9 pins) 2.0 or higher 1 x (or printing on network) 1 x (32 speed) 19" TFT flat screen set 96 dpi Mouse, keyboard and printer Operating system MS Windows TM 7 Professional or Ultimate (recommended) 32/64 bit Note: OS must be installed completely, including screen resolution, fonts and original video drivers. Note: FlowAccess V3 supports several languages, but only when the common rules for decimals and separators for the region are obeyed Installation instructions Notes: 1. It is strongly recommended to exit all applications before starting the installation. 2. Be sure to be logged in with all (administrator) rights to the computer before installing the software and the user account settings in Action Center must be set at Never notify. 3. Windows Installer version 3.1 (V2) has to be present on the PC before installing the FlowAccess V3 software. This Windows Installer can be downloaded from the website link as follows, -5F56-4A38-B838-DE776FD4138C&displaylang=en 4. Read carefully the Last minute instructions and the FlowAccessV3 instructions 3.1.xx on the DVD before installation. The latter file contains information about the code to be inserted during installation. Installation: 1. Start Microsoft Windows. 2. Insert the DVD-ROM in the CD drive. After a few seconds the Autoplay screen is displayed. 3. Click Open folder to view files. Windows Explorer is opened with the DVD drive selected. 5. Double click on Setup.exe in the right part of the screen. The Install Wizard is opened and starts to prepare the installation.. 4. Follow the instructions on the screen for the installation of the software. Note: Make sure to select a convenient directory for the data. 5. When the software is installed the FlowAccess V3 program is displayed in the Programs group on the start bar and the FlowAccessV3 instructions manual is placed on the desktop. 6. Open the program by clicking the left mouse button. The program window (shown in Figure 1.15) will be displayed. After login some extra screens are displayed for final definition (see chapter and 1.7.3).

16 FlowAccess V Software Update Introduction Before installing an update of FlowAccess V3 make a backup of your database (see chapter ). If one way or the other the installation was interrupted still a backup of the database can be restored. Warning: Before installation of an update, make a note of the current Matrix settings (if applied) and Peak detection settings, as these settings are lost! After installation of the update enter these settings into the updated program. Warning: If the sampler positions (coordinates) for your sampler were changed after installation of the previous version of FlowAccess V3, make sure to have a screen dump or note of these sampler positions. The positions installed in the upgrade have to be changed into these positions before the upgrade can be used for analysis (see chapter , Tray settings) Installation of an update Note: It is not necessary to remove the older version of FlowAccessV3 TM Installation: 1. Insert the DVD-ROM in the CD drive. After a few seconds the Autoplay screen is displayed. 2. Click Open folder to view files. Windows Explorer is opened with the DVD drive selected. 3. Double click on Setup.exe. The following message is displayed: Figure Click Yes to continue the setup. 7. The Install Wizard is opened and starts to prepare the installation. 8. Follow the instructions on the screen for the installation of the software. 9. Click Finish to close the Install Wizard Updating System Database Automatically any software patches are installed with the FlowAccessV3 program. The database has to be updated with these patches in the program by means of the DB Version Updation. 1. Open the FlowAccessV3 program. A message is displayed: Version conflict in database. Click OK. 2. The screen as shown in Figure 1.2 is opened automatically. Note: DB Version Updation can also be opened by the icon in the left pane, when the General Settings tab is active.

17 Chapter 1 Introduction to the FlowAccess V3 software 1-5 Figure Click Version Updation. The screen as shown in Figure 1.3 is displayed as an example. In this screen the FlowAccessV3 database can be updated to the required version Updating run files Figure Select FlowaccessV3 Database and click Apply Patches. The database will be updated (in the column Results updated is shown). Note: When SQL server 2005 is used start from 1 to apply the patched. When SQL server 2008 is used start from 133 to apply the patches. The displayed list is changed by entering the number and pressing Refresh. In the same way the databases for QAccess and FlowReport (if installed) can be updated by selecting the Module name. By means of the above screen also all run files can be updated to the current database version in order to view them correctly in PostAnalysis. Select All run files and click Apply Patches (this may take a while) and then Apply CLR Calculation. A screen is opened where FlowAccess V3.dll is to be selected (see Figure 1.4).

18 FlowAccess V3 1-6 Click Open and then confirm the updation. All run files are updated and recalculated. Figure 1.4 Also run files that were imported from another source can be updated to the current database version to be able to view them in PostAnalysis. Note: The run files cannot be imported from a network drive, but must first be saved on the PC. 1.5 When a PC is renamed after installation of FlowAccess V3 When the name of a PC is renamed by the customer after installation of FlowAccess V3 (e.g. when the PC was delivered by Skalar with a complete SAN ++ system), this PC name has to be changed in the.ini file of FlowAccess V3. 1. When FlowAccess V3 is started after renaming it seems that the system is hanging. However after a while (this may take up to several minutes!), an error message is displayed (Figure 1.5). Figure 1.5

19 Chapter 1 Introduction to the FlowAccess V3 software Press OK. Then the FlowAccessV3 INI file details screen is displayed (see Figure 1.6) Figure Change the Old PC name \SQLExpress into new PC name \SQLExpress. This action must be performed for each installed database (FlowReport, QAccess). 4. Press Update. After a while (the screen may say during this time that it is not responding) the following screen is displayed (see Figure 1.7). Figure Click OK. FlowAccess V3 can now be restarted normally.

20 FlowAccess V Reinstalling a database after renaming the PC When, for any reason, a database is (re)installed later (e.g. FlowReport.DB, FlowAccessV3. DB, FlowAccessV3.bak, etc.) via InstallDatabase, this installation will fail. The following message will be displayed: Connection string not initialized properly. Procedure to solve this problem: 1. First install SQL Server Management Studio, which can be found in the subdirectory SQL management in the FlowAccessV3 folder (see upper arrow in Figure 1.8). Figure Run the required program (32 or 64 bit, dependent on the computer system). 3. Then run the program FlowAccessV3RenameServer.exe which can be found in the FlowAccessV3 folder (see lower arrow in Figure 1.8). 4. The Server Details screen shows the computer name and the server name (see Figure 1.8). 5. Press OK. A message is then displayed (see Figure 1.9). Figure Select Yes. The PC system is closed and restarted. 7. Now the database can be installed via InstallDatabase, which can be found in the FlowAccessV3 folder. 8. Select InstallDatabase.exe to open the Welcome screen and click Next.

21 Chapter 1 Introduction to the FlowAccess V3 software Wait in the next screen (see Figure 1.10) until the (new) server name is displayed. 10. Enter the user name (sa) and password (Lobby@123). 11. Click Next. Figure Enter the login name (sa), login password (Lobby@123) and confirm the password. 13. Click Next. A message is displayed that the user already exists. Click OK. 14. In the next screen (see Figure 1.11) browse to the directory where the database to be imported is present. Figure Then select the path where the pre-defined database is to be stored. This is the location that was selected for the database during installation. 16. After a few seconds a message is displayed that the database is successfully restored. 17. Restart FlowAccessV3.

22 FlowAccess V Communication settings The communication settings for proper communication between the software program and the analyzer are set in the Communication Module, which is present in the directory where FlowAccess V3 is installed. A. Log settings Activate the registration of the communication in a log file. This registration is important for locating errors in the communication. Each time the program is started a file called Communication Module.log starts to register every step in the communication between the system and the program. The file is saved in the FlowAccessV3 directory when the program is ended, but is overwritten as soon as the program is started again. If any errors have occurred in the communication, this file can be viewed in Wordpad or Notepad or can be sent to Skalar for investigation. To activate the logging: 1. Open Windows Explorer and move to the directory where FlowAccess V3 is installed. 2. Double click on the file Communication Module.exe. The program is opened (see Figure 1.12). Figure Select Logging from the File menu or click on the Logging icon (1 st left) on the toolbar. The Information Logging dialog box is opened (see Figure 1.13). 4. Select Single file logging for the Log mode. 5. Check the options Read Packets and Write Packets and click OK. The dialog box is closed. 6. Close the Communication Module program. Figure 1.13 Important note: Always rename a log file that needs to be checked for errors in the communication before the FlowAccess V3 program is re-started.

23 Chapter 1 Introduction to the FlowAccess V3 software 1-11 B. COM port settings By default the program assumes COM port 1 of the computer is used for the connection with the analyzer system. If you use another COM port for the connection, this has to be changed in the Communication module 1. Open Windows Explorer 2. Move to the directory where FlowAccess V3 is installed. 3. Double click on the file Communication Module.exe. The program is opened. Select Properties from the File menu or click on the Properties icon (2 nd left) on the toolbar. The Communication Settings dialog box is opened (see Figure 1.14). Figure Click the arrow for the port and select the proper COM port from the list. Check also if the option Connect to instrument is selected. 5. Click Apply and then OK to close the dialog box. 7. Close the Communication Module.

24 FlowAccess V The Main Window When the software is installed a FlowAccess V3 icon is displayed on the desktop. Open the program by double clicking the left mouse button. The following screen is displayed (Figure 1.15). Figure 1.15 Main window Login To be able to enter the program for the first time the master user name and password has to be entered. The master user name is Administrator and the password is admin. Figure 1.16 Change password Warning: This password has to be changed into another (secret) password by the highest user (administrator) of the program to prevent unauthorized admission to the User Management window.

25 Chapter 1 Introduction to the FlowAccess V3 software Click on Change Password. The screen of Figure 1.16 is displayed. 2. Type the old password (admin) again. 3. Then type a new password and confirm the new password. 4. Select Change. A message is displayed that the password is successfully updated. 5. Select OK. The Login screen (see Figure 1.15) is displayed again. 6. Type the new password and select Login. The screen is closed. Only when the Administrator has logged in (the user name Administrator cannot be removed) the option User Management (see chapter ) is available from the Settings menu Restore a pre-defined system setup When FlowAccessV3 TM was ordered with a complete system, the system was already setup in FlowAccessV3 TM and a backup database was stored on the installation disk. Copy this database backup file (FlowAccessV3.bak) to a directory your hard disk. As soon as you are logged in a message is displayed whether you want to restore the Factory setup database or not (see Figure 1.17). Figure 1.17 Restore Pre-defined database If you select No the original empty database is used an you have to setup completely your analyzer system. If you click Yes a Database Wizard screen is opened (see Figure 1.18). 1. Browse to the directory where the pre-defined database was stored and select the pre-defined database. Figure 1.18

26 FlowAccess V Then select the path where the pre-defined database is to be stored. That is the location that was selected during installation for the database. 3. Click Next. 4. After a few seconds a message is displayed that the database is successfully restored. 5. Restart the program. The pre-defined system setup will be displayed in the Main screen when the System tab is active Path settings As soon as a choice is made to use the original database a new message pops up (see Figure 1.19). Click OK. The Preferences screen is displayed with the Path settings all set on C:\ (see Figure 1.20). Figure 1.19 Click the Browse icon and select for each item the subdirectory of FlowAccessPost. The location of FlowAccessPost was selected during installation. Click OK to close the screen. Figure 1.20

27 Chapter 1 Introduction to the FlowAccess V3 software Main window items The screen displays the main window with a menu bar, a toolbar and a twopane window. The left pane shows the main parts of the program: System, Data Processing, Reports, Quality Control and General Settings. The System tab (see chapter 2 and 3) is active (and the sub items displayed in the upper part) when the program is opened. The toolbar displays the shortcut icons for the system. The right pane displays the setup system(s). Click on Data Processing (see chapter 9) to make that tab active. The right pane displays the performed runs. Reports and Quality Control are optional programs within FlowAcessV3 and are described separately. Click on General Settings to make that tab active. The right pane displays the current settings for the interface and the current settings for the preferences (see Figure 1.21). Figure 1.21 By double clicking on a setting either the Interface Settings screen (see Figure 1.22) or the Preferences screen is displayed (see Figure 1.28), where changes in the settings can be made. Both screens can also be opened from the icons in the upper left screen or as an option from the Settings menu. De DB version Updation option (described in chapter 1.4.3) is also displayed in the left upper part of this screen. The menu bar displays the following main menus: File, Settings and Help.

28 FlowAccess V The File menu The File menu contains the options: System, Analysis, PostAnalysis, Reports, Update Configuration Settings, Quality Control, Log Off and Exit. System (see chapter 2), Analysis (see chapter 3) and PostAnalysis (see chapter 9) of the menu are also accessible by using shortcut keys or as icons in the left upper part of the screen, when the tab is active. Reports and Quality Control can only be used if they are installed Update Configuration settings When this option is selected the INI file details screen is displayed with database information. The program cannot run when conflicts are shown in this screen (see also chapter 1.5 when a computer was renamed) Log Off When this option is selected the user is logged off (after confirmation) and the User Login dialog box is displayed. When this dialog box is closed by means of x without logging in, most program items (except Update Configuration Settings) are unavailable. Also when Cancel is pressed, most program items are unavailable. Log Off in the File menu is changed into Log On, when selected the User Login dialog box is displayed again The Settings menu The Settings menu has six options: Interface Settings, User Management, Preferences, 21 CFR part 11 support, Database Management and Search in Runs. Except for the last item, all items (or sub-items) of the menu are also accessible by using shortcut keys Interface Settings When selecting this option the following screen is opened: (see Figure 1.22). Figure 1.22 Interface Settings

29 Chapter 1 Introduction to the FlowAccess V3 software 1-17 In this screen all items concerning the interface are set. Select for each item the total number that is physically present on the system that will be defined. Note: Interface Settings can also be opened by the icon in the left pane, when the General Settings tab is active. First select the type of interface that is used on the system and then the configuration of your system. - System Controllers Up to 3 system controllers can be present. - Digital Detectors A maximum of 8 digital detectors can be connected to the interface. - Density Meters One density meter can be connected to the interface. - Heaters A maximum of 8 heaters can be connected to the system controllers. - External Valves This option is for future use. - Sampler Valves This option is for future use. - Sampler Up to 3 samplers can be connected to the interface. - Analog Detectors Up to 16 analog detectors can be connected to the interface. - Rinsing Valves Up to 6 rinsing valves can be connected to the system controllers on the interface (2 per system controller) - External Rinsing Valves This option is for future use. - Channel Valves This option is for future use User Management Only the administrator is able to open this option. When this option is selected the following screen is opened: (see Figure 1.23) with two panes. The settings for the Administrator are displayed and cannot be changed. Figure 1.23 User Management The left pane shows the user groups; the right pane the user details. In this screen all parts of the user management are set.

30 FlowAccess V New Group New user groups can be created. When the button is selected, the User Group Settings screen is displayed (see Figure 1.24). Figure 1.24 New Groups Enter the User group name and press Add. The screen is closed and a new group is added to the list. - New user New users can be created in any group that was created. Selecting this option opens the User Master screen (see Figure 1.25). Figure 1.25 Enter the User ID, User Full Name and User Group, where the new user is placed. Select the date that the password will expire. Optionally enter the address. Press Save to save the settings and to close the screen. Note: Automatically the User ID is the user password, when the new user logs in for the first time. A message is displayed that the password is empty and that the password has to be changed. When clicking OK the Change Password screen is displayed. The user has to change the password to his own (secret) password as shown in Figure Reset Password When this option is pressed the password for all users will be deleted and a new password has to be entered next time the user logs in.

31 Chapter 1 Introduction to the FlowAccess V3 software 1-19 Figure 1.26 Change password - Edit user First select the name of the user to be edited from the list and press Edit User. The screen of Figure 1.25 is displayed with the details of the user and the changes can be made and saved. - Delete user Select the user group of the user to be deleted from the list and press Delete User. The User Delete screen pops up with all users of the selected group. Select the user to be deleted and press OK. The deletion has to be confirmed before the user is deleted. - User Rights The User Rights can be set for every individual user or group. Press the button to open the User Rights screen (see Figure 1.27 for an example). Figure 1.27 User Rights

32 FlowAccess V Preferences Select the User Group and User ID before settings the rights. If all rights will be admitted press Select All. All other rights are admitted or refused by clicking the tick boxes. Press OK when the required rights are set. Note: Preferences can also be opened by the icon in the left pane, when the General Settings tab is active. When this option is selected the Preferences screen (see Figure 1.28) is displayed with six tabs: Path Settings, Color Settings, File Settings, Other Settings and Mail Settings. - Path Settings, In this tab are set: - Run file storage path - Import run file path - Export run file path - Log file path After installation of the software the path is set to the root. The program asks to store your data on a different partition of the hard disk, but not in the FlowAccess directory. Select the browse button and select the required path (see also chapter 1.7.3). Figure 1.28 Path settings - Color Settings (see Figure 1.29), In this tab the color settings for max. 20 modules can be set for: - Real time Colors - Connection Colors - Group Colors

33 Chapter 1 Introduction to the FlowAccess V3 software 1-21 To change the color: click on the color to be changed, select the required color from the pop-up screen and click OK. Figure 1.29 Color settings - General Settings (see Figure 1.30) Figure 1.30 General settings In this tab several option of the run can be defined: - Run file format: the default format is SKALARyyyyMMdd[A][9] - Grid style (only in system setup and active system) - Show the Communication Module - Show Message alert

34 FlowAccess V Overlapping runs included - Execute Other apps - Allow other apps to access The Run file format can be changed and the format of the new output can be checked by clicking Check. The new output will not be showed if invalid. A grid is displayed in the screens of System setup and Active system when the option is checked. The Communication Module (see chapter 1.6) is displayed in the taskbar when the option is checked and an Active system is open. Message alert is displayed when this option is checked. This means that actions (e.g. of the sampler) are displayed shortly on screen. Allow to run in overlapping mode can be used in the case of large workloads. When this option is checked a new analysis can be started before the previous analysis is ended. This saves the retention time of the module. For explanation of this feature refer to chapter 8.7. Execute the external application is used to execute external applications to be used with FlowAccess. For example to export results to a LIMS system. This feature requires extra user defined macros and is only available on request. After the analysis this option is available from the File menu in PostAnalysis, when checked here (see chapter 9.2. The application settings have to be defined (see Figure 1.31 as an example). Figure 1.31 Application settings Allow access of an external application can be used to import tables from an external application to be started immediately in FlowAccess from this application. This application requires user defined macros and is only available on request. - Other Settings (see Figure 1.32) Hardware setup: The default pickup speed and dispense speed for a 10 ml diluter, when making working standards, are displayed. When another diluter volume is used these settings must be changed. The settings will be provided by Skalar. Warning: This speed can be changed, but this should only be done for a good reason.

35 Chapter 1 Introduction to the FlowAccess V3 software 1-23 The default margins for printing the results can also be changed here. The standard table columns like Identity, Weight, Comments and External dilution factor, which are selected by default, can be deselected from being displayed in the sample table. Figure 1.32 Other settings - Mail Settings In this tab options can be enabled and the details entered (not yet available).

36 FlowAccess V CFR part 11 This item is only available to those with the required user rights. This item has three options: Settings, Audit Trail and Login Settings. - Settings In the CFR Part 11 Settings screen (see Figure 1.33) General settings as Enable Audit Trail, Enable Electronics Records Support and enable Electronic Signature Support can be selected. Enable Audit Trail is used to track the login and actions of a user inside the program. The log file of the Audit Trail is viewed in the Audit Trail window that can be opened from the 21CFR part 11 menu. Electronic Records Support is used to rename a run file when the file is edited in the table or the peaks. The original run file is saved under the original name. Electronics Signature Support is used for the supervisor to approve a run file. This feature is not yet implemented. Reason resources can be created that must be used when editing is performed in the run file. For example Editing peaks, Analysis Time-out etc. When a reason is typed, click Add to add it to the list. A reason can be deleted from the list by selecting the reason and clicking Del. - Audit Trail Figure 1.33 CFR 21 Part 11 All CFR transactions are displayed here (see Figure 1.34), when Enable Audit trail is checked. Selections for the display can be made in dates, users, actions and parts of the system. Press Show to show the selected actions. The actions can be printed and exported. The displayed actions can be archived by pressing Create Archive. An Archive can be opened by pressing Open Archive.

37 Chapter 1 Introduction to the FlowAccess V3 software 1-25 Figure 1.34 Audit Trail - Login Settings When this option is selected the screen as shown in Figure 1.35 is opened. Here the general settings for all users are defined. Figure 1.35 Login Settings Password length: The minimum length of the password. By default the password length is set on at least 4 characters. The minimum password length is selectable between 4 and 50 characters. Idle Log off time: The waiting time until the user is logged off automatically if the program is not used. This time can be set between 1 and 99 minutes. Default is 60 minutes. Login Failure attempts: The number of times the user may enter a wrong password before the program is aborted. This can be set between 1 and 99 attempts. Default is 3 times. Select OK to accept the settings.

38 FlowAccess V Archiving Settings When this option is selected the screen as shown in Figure 1.36 is opened. Figure 1.36 Here a schedule can be entered for archiving the actions in the Audit Trail. The Archive can be opened from the Audit Trail screen Database Management When the option is selected a screen is displayed (see Figure 1.37) with two tabs: Backup Database and Compact Database. Select a path for the backup to be stored. Click OK and a backup of the current database is made and placed in the selected directory. Figure 1.37 Backup Database In Compact Database (see Figure 1.38) the database to be compacted can be selected (if present). Click OK to compact the selected database. Figure 1.38 Compact Database

39 Chapter 1 Introduction to the FlowAccess V3 software Search in Runs To ease the search for certain samples an option is available to filter the search on the name of the sample. Select Search in Runs to open the screen (see Figure 1.39). Figure 1.39 Search in Runs Enter the identity of the sample to be found or a part of the identity and press Search. The program searches in the existing run files and places the list of files that matches the criteria in the screen. Select a file from the list and press Open to view the file The Help menu The Help menu shows the online help for the use of FlowAccessV3 and the info about the version of FlowAccessV3 and the computer. Both items of the menu are also accessible by using shortcut keys.

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41 FlowAccess V3 CHAPTER 2: SYSTEM SETUP Preface: This chapter gives detailed instructions for a complete system setup.

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43 Chapter 2 System setup SYSTEM SETUP 2.1 Introduction Before an analysis can be started the analyzer system has to be defined in all its aspects. If Skalar installed FlowAccess V3, the system has been defined already by a Skalar staff member. In this case just check the system setup. If any changes are required, we recommend you to follow the instructions in this chapter. If you install FlowAccess V3 yourself, follow the instructions in this chapter for the setup of your system. After Login the screen as displayed in Figure 2.1 will be displayed (when no systems were already defined). Figure 2.1 Note: A system is usually setup only once. The setup of a system is necessary for the software to know which hardware components are used on the system. The chemistries on the system are defined in the Active System screen. If a chemistry on the system is used for several methods or ranges, it may be easy to create more systems, i.e. for water, soil and fertilizer. However, it is also possible to replace the method on the chemistry in the Active System screen. In the upper part of the left pane four icons are displayed: New System, Edit System Delete System and Open for Analysis icon. These items

44 FlowAccessV3 2-2 can also be selected directly from the toolbar or by selecting System from the File menu. When no systems are present the icons Edit System and Delete System are gray. In the case that already systems are defined, these systems are displayed in the right pane. Information about the hardware components can be checked in the Interface Settings. Click New System to create a new system. In the case that no system was yet defined the Interface Settings screen is displayed to set the hardware information of the system. Enter the required information, see chapter for details. Click Save and Continue. A message is displayed that the settings are successfully updated. Click OK to close the screen. The Hardware Setup screen will be displayed (see Figure 2.2). Figure 2.2 The screen is divided in three panes: - The left pane contains four tabs with all hardware parts for your system; the active tab shows the selectable items. - The center pane will show the hardware that was selected in the system. Figure 2.2 shows the middle pane with grid (set in the tab File Settings of the Preferences screen, see chapter ). - The right pane contains five tabs with all the connections that can be made with the selected hardware and one tab with the properties of a selected item. The active tab shows its possible connections (this is dependent on the Interface Settings, see chapter ). With the aid of the toolbar items the system is compiled. The Add and Remove buttons are used to add and remove the hardware items; the Connect and Disconnect buttons are used to connect and disconnect the hardware parts. Up to 3 systems can be connected to the interface, each in its own tab.

45 Chapter 2 System setup Setup of the system A system is defined according the physical layout of the system on the laboratory table Adding the hardware Note: Make sure that the Add button is selected before the hardware is selected to be added. When any hardware has to be removed first select the Remove button and then click on the item to be removed. 1. Select the Sampler tab in the left pane. This tab has two sub-tabs: Samplers and Accessories. The Samplers sub-tab will be active. Then all available samplers are displayed. 2. Click the Add button (a + is added to the pointer) and click on the picture of the sampler that is physically present on the system. The selected sampler is then displayed with one needle in the center pane. 3. Select the Accessories tab. The possible accessories (diluter, extra valves and extra needles) are displayed. Note: The valve required for the diluter is part of the diluter and is not seen as an extra valve. 4. Click on the accessories physically present on the sampler of the system. The items are added to the sampler in the center pane. Note: The SA1053D sampler is a special: 2 needles and a diluter, which however cannot be used simultaneously. Select the SA1053D when one needle with or without diluter is used (right needle up or removed). When the dilutor is used, add the dilutor. For use of 2 needles (no dilutor) the SA1050 is selected. This type selection defines the matrix of the sampler and thus the available racks in the Edit trays screen (see chapter ) and the Table Entry screen (see chapter 4.5.1). 5. Then select the Module tab in the left pane. The options are displayed (the option module valve is not yet available). 6. Click on the picture of the module (the module part in the center pane turns white) and click once on the white area for each module that is physically present on the system. If more than one module holder is present, a gap can be held between the modules on the different module holders (see Figure 2.3). 7. If heaters are present, click on the picture of the heater and click in the module in the center pane, where a heater is used to add the heater to the module. 8. Select the Detectors tab. This tab has two sub-tabs: Digital detectors and Analog detectors. Both types can be present simultaneously on the system. The Digital detectors tab will be active. 9. When digital detectors are present: Click on the picture of the required digital detector (dual or matrix) and click in the white area behind the module where a digital detector is present. When a dual detector was selected, automatically two detectors are placed behind the module and the next module. 10. When analog detectors are present: Select the tab, click on the picture of the required analog detector and click behind the module where the detector is present. (analog detectors are seen as single channel). 11. Select the Pumps & Valves tab. Click on the picture of the pump and then click on the white area in front of the first module. One pump for one module holder. If a second module holder is present, click in front of the first module in the second module holder.

46 FlowAccessV Click on the picture of the rinsing valves and then click on the white area in front of the pump where the valves are placed (usually two per module holder). Note: The options External rinsing valve and External valve are for future use. 13. Now click on the pointer and then on the pump. Then click on the arrow up icon to extent the pump over all modules in the module holder. Do the same with a second pump (if present). 14. Then click on the first rinsing valve to extent the valve over a second module. Do this for each rinsing valve present on the system. 15. Click twice with the pointer on module 1. Type a name for the module. Repeat this step for all modules. 16. Now this software system setup represents the physical layout of the analyzer system on the laboratory table (see example in Figure 2.3). Figure Connecting the hardware Then the hardware has to be connected. Note: Make sure to select the Connect tool each time a new part has to be connected. Always when the Connect button is pressed a + and tray is added to the pointer. As soon as then one of the parts to be connected is selected the + and tray is changed into an S until the connection part is selected. A Mismatch message is displayed when a connection is not possible. 1. Select the Sampler Connector tabin the right pane. 2. Select Connect in the toolbar. Click on the sampler in the center pane and then on the Sampler connector 01 in the right pane. The sampler and the connector get a pink lining to show that they are connected. If a second sampler is present connect this sampler to connector Select Connect. Click on the needle and then subsequently on each module that is connected to the needle. The left front of the modules get a color lining with the same color as the needle to show the connection.

47 Chapter 2 System setup Select Connect. Click on the pump and then subsequently on each module connected to the pump. The pump and the modules turn pink to show the connection. 5. Select the Detector connector tab. 6. Select Connect. Then click on module 1 and then on the detection head behind the module. Repeat this step each module. To show that they are connected the end of the module and the detector get a yellow lining. 7. Select Connect. Then click on the detection head behind module 1 and then on dual connector 1,2 in the right pane. When the detector is a dual detector also module 2 is automatically connected. Repeat this step for all other modules. When a module with a matrix detector is present, only one module can be connected to a dual connection in the right pane. A density meter is connected to an RS connection (physically Sampler 2 on the interface). An analog detector is always connected to a single connector. The connectors get a pink lining to show that they are connected. (For a single channel connection (see chapter ). Note: Always first connect the modules to the detection head before connecting the detection head to the connector in the right pane. 8. Select the System controller tab. 9. Select Connect. Click on the pump in the center pane and then on the pump icon in the right pane. The icon turns green to show the connection. 10. Select Connect. Click on the first heater on the module and then on the Heater icon in the right pane. The icon turns green and the heater on the module gets a pink color. Repeat this step with the second heater. This heater also turns pink. Figure 2.4 Connecting the hardware Note: The digital detectors are already connected and are not separately connected to the system controller. This connection is for analogue detectors only.

48 FlowAccessV Select the Rinsing Valve tab. 12. Select the Connect. Click on rinsing valve 1 and then on the connector 01. The rinsing valve turns pink and the connector gets a pink lining to show the connection. Click on rinsing valve 2 and then on connector 02. Also this valve turns pink and the connector gets a pink lining. 13. Select the Pointer in the toolbar. A system with 2 samplers may now look like Figure 2.4. Note: The Other Valves tab is for future use. 14. By means of the Properties tab all parts of the setup can be viewed with their status. The status of the pump, heater, rinsing valves and detectors in the different modes (default, standby and online) as well as the settings for the sampler can be set here as a standard for the system. The status settings are explained in chapter Note: The status of the pump, heater, rinsing valves and detectors in the different modes (default, standby and online) can be set also in the analysis window. 15. Click Save. Type the name of the system in the displayed dialog box and click Save again. The box is closed. Note: If not all required connections were made a dialog box with warnings about the missing connections will be displayed. Make a note of the missing connections and close the box. Click Cancel in the dialog box and first connect the missing connections. Then save the system again. 16. Click Exit to close the Hardware Setup screen. A confirmation is asked before the screen is closed. The Main screen will be displayed with the icon of the system just made Single channel connection When a setup is made with one channel and a digital 2-channel detector, first click on detector side A to and then on Digital 1,2 (see Figure 2.5). When detector side B is connected to the interface, this will result in changing the channel. So B will become A and reverses the real time. Figure 2.5 Connection of a single channel

49 Chapter 2 System setup Status settings Click the Properties Tab. By means of this tab the status of the pump, heater, rinsing valves and detectors in the different modes (default, standby and online) are set here as a standard for the system. 1. Click on rinsing valve 1. In the upper right pane the properties of the rinsing valve is displayed for each mode (see Figure 2.6). Figure 2.6 Status settings 2. Select the properties (on/off) for valve 1 in each mode. 3. Do the same for each valve present on the system. 4. Select pump 1. Select the properties for each mode of the pump. Do the same for each pump on the system. 5. Select a heater on the system and select the properties for each mode. The properties are valid for all heaters. 6. Select the detector behind channel 1 and select the properties in each mode. If the detector is a dual channel the properties are also valid for channel Select the detector behind the next channel and select the properties in each mode. Do the same for each channel in the system. In the case of a matrix detector select the required measuring channel. Note: For the detectors high voltage can be selected for applications that are measured on a wavelength between 300 and 400 nm to increase the light intensity. For measurements on wavelengths higher than 400 nm it is advised to keep the lamp on low voltage for a longer lifetime. 8. Select sampler 1 on the system. The sampling properties are displayed. Change the default settings if required. For explanation of the sampler settings refer to chapter If more than one sampler is present do the same for these samplers. 9. When all properties are set, the connections can be viewed in an overview by selecting Connection overview from the Settings menu.

50 FlowAccessV3 2-8 Figure 2.7 shows an example with a SA1050 sampler, four channels, two heaters and two digital detectors. Figure Continue with step 15 of chapter The system can now be opened for further setup in the Active System screen. Select the system in the Main screen and click on Open for Analysis in the left pane or select the system and click the right mouse button to open a menu and select Open for Analysis (see Figure 2.8). Figure Edit a system The system can be edited as required. 1. Select the system to be edited in the right pane and then Edit System in the left pane (or right click on the system and select the item from the menu). The system will be displayed like in Figure 2.4.

51 Chapter 2 System setup Delete a system 2. Edit the system as required and click Save. 3. Click Exit. Click Yes to confirm the exit to the Main screen. A system can be deleted from the FlowAccessV3 program. 1. Select the system to be deleted in the right pane and then Delete System in the left pane (or right click on the system and select the item from the menu). The following screen is displayed (see Figure 2.9): 2. Click Yes to confirm the deletion. Figure 2.9

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53 FlowAccess V3 CHAPTER 3: ACTIVE SYSTEM Preface: This chapter gives detailed instructions about all aspects of the active system

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55 Chapter 3 Active system ACTIVE SYSTEM 3.1 Introduction The Active System window can only be opened after a system has been setup. The system setup is the basis for the setup of the analyses. The Active System window is used for: - Definition of the chemistries on the modules - Definition of the group(s) of chemistries for analysis. - Preparation of system table(s) and/or sample table(s). - Setting the status for detectors, rinsing valves, heaters and pumps (or changing if the status was set in the Setup window) in the different modes. - Setting the settings for the sampler (if not set in the Setup window). - Setting and enabling the time programming (automatic start-up and shutdown) - Viewing the status of the hardware. - Starting, stopping, viewing and controlling the analysis runs. The Active System window is opened from the main program window. Select the system in the Main window and then Open for Analysis in the left pane The Active system window displays the selected system. An example of the Active system window is shown in Figure 3.1. Figure 3.1 The Active system window

56 FlowAccess V The Menu bar The window displays the system as setup in Figure 2.4. No group name is displayed as no methods and groups are yet defined. The window displays a menu bar with 6 main menus: File, Analysis, Table, Methods, Instruments and Other Settings. The Toolbar displays 14 easy access buttons where most options of the main menus can be accessed. A dashboard is displayed at the right side of the screen with the current status of the system. During analysis this info is updated. A system setup with more than one group or with more than one sampler, can be run with these groups or samplers independently. Time programming can be used for shutdown (one time program per system). In this case the program shuts down when the last run is finished. Note: When time programming is used for automatic start-up, only one group can be started The File menu The File menu has two options: Exit and Shutdown and Exit (see Figure 3.2). Exit can be used to close the Active System window to return to the main window. Exit can Figure 3.2 also be reached from the toolbar. The window can also be closed by close (x) box of the window. Shutdown and Exit is used to shutdown the system and to close the Analysis window to return to the main window The Table menu The Table menu has two options: New Table and Open Table (see Figure 3.3). New Table is used to make a sample table for all samplers available in the system (see chapter 4.3). Edit Table is used to open any existing Figure 3.3 table of all samplers available in the system. Table can be clicked on the toolbar and then on the displayed sampler, but then only tables can be created, loaded, edited and removed for the selected sampler The Analysis menu The Analysis menu has two options: RealTime and Results (see Figure 3.4). RealTime opens the screen where the run can be viewed graphically. RealTime is explained in chapter 8.7 Figure 3.4 and 9.2. Results opens a screen where the incoming results are displayed. Results is explained in chapter 9.3.

57 Chapter 3 Active system The Methods menu The Methods menu has three options: New Method, Edit Method and Custom Method (see Figure 3.5). New Method is used to define a new method. This method is automatically assigned to the first module unless another module is selected. When a Figure 3.5 method was already assigned to the module the new method replaces the method of the selected module on the system, but the original method will be available when a method has to be added to a module. Edit Method is used to edit an existing method (attached to a channel). It is possible to view also the methods of any other system setup in FlowAccess V3 but then the methods cannot be edited, only deleted. Only the methods of the current system can be opened and edited. Custom method is used to display defined methods that are not assigned to a module. New methods can be defined and existing methods edited and deleted The Instruments menu The Instruments menu (see Figure 3.6) has two or three options: Time Program and Master/Slave settings are always available. Barcode settings is only available when installed. (Barcode Settings is optional and used to identify the identity of the sample by scanning the barcode on the sample tube. De Barcode Scanner is explained in a separate manual). Time Program is used for programming an analysis with automatic start-up and shutdown. Time Program has four submenus: (see Figure 3.7) - Time Program Entry is used to setup the time program. - Time Program Selection is used to select the type of time program and the name of the used time program. It can be enabled here as well. - Enable Time Program is used to activate the selected time program. Figure 3.6 Figure Set secondary time program is used to enable a second time program to be enabled after the first time program. Time Program is explained in chapter 7.4. Master/Slave settings When selecting this option the Master/Slave screen is displayed (see Figure 3.8). In this screen a channel can be set as slave of another channel on the system. The module name is selected from the drop-down menu. The channel must have a link to a master channel and the channel has to be synchronized with the master channel. For the slave channel no calibration may be selected. The detection of the slave takes place on time basis.

58 FlowAccess V3 3-4 The slave channel has to be synchronized with the master channel. By default the synchronization mode is set on automatic, but this can be changed into manual. When manual is selected a delay time for the start of the slave channel can be entered The Other Settings menu 3.3 Toolbar Figure 3.8 Master/Slave settings The Other settings menu has three options: Save Comments, Analysis Information and System Information. Save Comments is used to save or clear comments. Analysis Information is used to Figure 3.9 retrieve information about the analysis. This option is only valid during analysis. System Information is used to retrieve information about the hardware on the system. For example the total working time of the pump. This option is only valid when the program is connected to an analyzer. The toolbar contains the shortcut buttons for most parts of the options of the main menus. Click once on a button to make it active. Status: To set or change the status of the system Remote Control: To control the settings of each part of the system in the different status Table: Method: Group: To create, load, edit or remove a table attached to the selected sampler To create, load, edit or remove a method attached to a chemistry of this system To create, load, edit or remove a group of chemistries connected to the active system

59 Chapter 3 Active system 3-5 Analysis: To select a group for analysis Baseline: To select a group to display the baseline in Real time Start: Stop: To start the analysis To stop the analysis Result: To open the Results screen Realtime: To open the Realtime screen Pointer: To select the mouse pointer Exit: To close the screen 3.4 Status settings Control When a system controller is present the system can be set in three different statuses: Default, Standby and Online. The status can be used as desired by the user of the system. For each status the hardware settings can be set in the System setup. In the Active System screen the Remote Control button is used to set the settings for the separate parts of the hardware in each status. The Active System screen is always opened in the Default status, when opened for the first time (which means that the red traffic light is on). When the Active System was closed in another status, the window will open in the same status as in which it was closed. The status is also displayed in the lower right pane Changing the status 1. Click on the Status button. A dialog box is displayed with the question: Do you want to change the System Status? and the current status and the new status. 2. Click Yes to switch to Standby. The traffic light is switched to yellow. 3. Click again on the Status button. Again the dialog box is displayed with the question: Do you want to change the System Status? and the current status and the new status. 4. Click Yes to switch to Online. The traffic light is switched to green Changing the settings in a certain status: 1. Click on the Control button. 2. Click on the hardware on the analyzer that has to be set in a different setting. A dialog box is displayed with the current setting of the hardware (On-Off for the pump, heaters and detectors; water-reagent for the rinsing valves). 3. Change the setting if desired by clicking on the current setting. The setting changes. Remember that all hardware connected to the same (part of the ) power strip is changed. 4. Click OK. The change is displayed on screen by a change in color on the hardware (yellow for the detectors; green for the rinsing valves, pump and heaters).

60 FlowAccess V3 3-6 When a system controller is used on the system with automatic start-up and shutdown, it is logic to set the settings of the hardware in the system setup. In this case the different statuses are used to prepare the system for the analysis. If no system controller is present on the system the user has to switch on all the hardware by hand. No control is possible in the software. Note: Remember that the system needs some time to start up. This means that a waiting time has to be set between the different status, when an automatic start-up is used. For the settings of an automatic startup see chapter 7.3. Note: When a system controller is present, an analysis can only be started when all necessary parts of the hardware are switched on. When not all parts are switched on, it is not possible to select the channels for analysis. Only when a rinsing valve is switched off (for example when this valve is not necessary for the reagents of the run to be started), still a selection can be made and in that case a message is displayed when the analysis is started Changing the sampler settings The sampler settings can be controlled as well, also without system controller. These settings are independent from the status. Click on the Control button and then on the sampler. A dialog box is displayed with the current settings of the sampler (see Figure 3.10). These are the default settings or the settings as set in the hardware setup. At the right side of the screen are four options available: Tray settings, Needle height settings, Pre-dilution utility and Drift cup details. Note: For samplers without diluter the option Pre-dilution utility is not available. The sample time, the wash time, the airtime, delay time, extra air time and the number of pickups can be changed (see Figure 3.10) for all types of samplers. Figure 3.10 Sampler settings Sample time: The time the needle draws up sample from the sample cup or standard cup

61 Chapter 3 Active system 3-7 Wash time: The time the needle is in the wash vessel between two sample pick-ups. This wash time is including the extra air time and the delay time. Air time: The time the needle draws up air to create an air bubble between samples and washes. Delay time: The time that the needle is in the wash vessel before the needle draws up extra air. This time is subtracted from the wash time. Extra air time: The time that the needle draws up extra air after taking a sample. This time is subtracted from the wash time. Note: The minimum wash time must be higher than the extra air time + delay time + the time needed for the sampler to reach the next sample cup. Note: The minimum wash time for SA1100 is 25 seconds due to rotation time of the cup plate. Note: At this moment it is not yet possible to change the liquid by means of the Change button. Flush the diluter if desired. With each click on Flush the diluter is flushed. Note: The Flush option can t be used when a SA1070 sampler with diluter is used. The sample pick-up can be tested. In Test, enter a cup number and click Go. The sampler takes a sample from the selected cup. The Pause and resume part is only available during a run. If required, the sampling can be resumed from another cup then where was paused. To achieve this enter the required resume cup number. Check the option Add Drift and wash cups after the break if the run has to be resumed with a drift and a wash Tray settings When this option is selected a screen with all possible racks for the selected sampler is displayed and their short ID character (see Figure 3.11). Figure 3.11

62 FlowAccess V3 3-8 Note: When the used trays on the sampler are not present in this screen, the trays must be replaced by selectable trays. A tray can be selected and when clicked Next the details of the rack are displayed (see Figure 3.12). This screen defines the layout of the tray and the calculated positions of the cups in the tray and of the trays on the sampler. Do not change these positions unnecessary as changes could cause the needle to crash when trying to sample from the tray! Figure Needle height settings When a sample or standard is diluted the sample is mixed. This is performed by picking up some air and sample and pushing air and sample back into the sample. The height for pickup of the sample and the height for the air pickup can be selected here (see Figure 3.13). The max. air pickup heights are: SA , SA , SA In case of large sample take-up the pump can be set to always On when the option is checked. This option should also be checked when working standards are made and a top stirrer is present. Figure 3.13 Note: This feature only works during analysis, not when using the command Cup position to sample once in Sampler Settings.

63 Chapter 3 Active system Pre-dilution Utility Note: Before this utility can be used the configuration of the sampler must be known. This can be achieved by preparing (and saving) a small table for the sampler, as when a table is prepared the software asks for the number and type of racks that are present on the sampler. Refer to chapter 4.5 to prepare a table. Figure 3.14 This utility is often used to prepare samples with a high dilution factor, but can be used for any dilution. The sampler is used simply as a diluter station. This dilution factor can be any factor larger than one. The actual dilution is carried out in steps of factor Select Pre-dilution Utility to open the screen as shown in Figure Press New to prepare a new dilution table or Load to load an existing dilution table. The Table definition screen is opened. (see Figure 3.15). Figure 3.15

64 FlowAccess V Enter the location of the sample to be diluted, the dilution factor and the target cup. It is convenient to place the target cup on the position from where the cup later will be analyzed (after creating a sample table for analysis). The Intermediate cups are filled in automatically when the file is saved and started and will be located in the last positions of the last available rack. Make sure to place enough sample tubes for the intermediate dilutions. Figure 3.15 shows an example for the table. The number of intermediate cups can be calculated when keeping in mind that each dilution has a max. factor of 10. So 100x needs one intermediate cup, 101x -1000x need two intermediate cups. 4. If required enter the number of mixing times during the dilution. 5. Press Save or Save and Start. Type a name for the table in the popup screen and press Save. The screen is closed. 6. When Save and Start was pressed the sampler starts immediately with the dilutions. The screen can be opened again during the dilution (by pressing Control and then on the sampler) to view the locations of the intermediate cups. When Save was pressed the screen as in Figure 3.14 was displayed with the filename of the table. The table can then later be selected and loaded. 7. When the table is finished the normal sample table can be loaded and the sampling started. Note: Make sure to replace the tubes of the intermediate locations and replace them with empty tubes in the case that post-dilutions are required Drift Cup details When long runs are applied it may be convenient to reserve more than one drift cup for the drifts. This option takes care for this (see Figure 3.16). Type the locations of the cups that are filled with drift liquid, check the box to activate the settings and press Save. During the run the sampler will take the drifts alternately from the selected cups. Figure 3.16 Note: When this screen is set on active, the settings in this screen can overrule the settings for the drift as set in the Table Wizard (see chapter 5.5.1). When a table is saved, and different Drift positions were set in the Table wizard, the program asks if the settings for the drift must be update to the settings that are set in this screen.

65 FlowAccess V3 CHAPTER 4: TABLES Preface: This chapter gives detailed instructions about all aspects of the tables for the analysis run.

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67 Chapter 4 Tables TABLES 4.1 Introduction Before a run can be started, a table has to be created and attached to the sampler. Table is accessible in two ways. 1. With the Table menu, tables can be setup for the samplers available in the system and tables edited for all samplers available in all systems. The tables that are setup in this way are not automatically attached to the sampler in the displayed system. The table must be attached after creation by means of the Table tool. 2. When clicking the Table tool on the toolbar and then on the sampler in the screen, a table for the active (displayed) system can be created or an existing table for the active system can be loaded, edited or removed. 4.2 Sample table A sample table contains the actual samples with their identification and other relevant factors (manual pre-dilution factor, weight factor) and automatic pre-dilution factor and dilution location, if a diluter is present. Two options for working with a sample table are possible: 1. A table comprising the full run (samples, standards, drifts and washes). 2. A table with only samples. In this case the calibration of a previous run is necessary for the calculation (see chapter 4.3.3). When using the Table menu first information is required for which sampler in your system the table will be created (important when different samplers are present in the system). Also the used racks on the selected sampler (in some cases more than one configuration is possible) have to be checked before the Table Entry screen is opened (see chapter 4.3). When using the Table tool a sampler has to be clicked. Then the used racks have to be checked before the Table Entry screen is opened (see chapter 5.3), when no table is yet created. If the sampler uses different types of racks, the racks can be selected within the Table screen.. Creation of sample tables is explained in chapter 4.5.

68 FlowAccess V The Table Entry screen This screen shows an empty table, when a new sample table is to be created (see Figure 4.1). T W D S1..99 U DI Figure 4.1 The Table always contains columns for position and type and if a diluter is present on the sampler (and defined in the system setup), the pre-dilution factor and pre-dilution position. Columns for identification, weight, comments and external pre-dilution factor may be present when checked in Preferences, Other Settings in the Main screen. The positions on the sampler are defined as follows: for the standard tray ST followed by the position (e.g. ST1, ST2 etc.), for the sample trays the tray ID followed by the number in the tray (e.g. A1, C4, etc.), for the wash Wt. Many different sample types are available. Here follows a list of available sample types with their explanation: A Tracer is the first peak of the Analysis run. Usually a known high standard is used for this first peak to ensure exceeding the start value. The Tracer is not included in the calculations. A Wash is a rinsing liquid sample taken from the wash tube. It is used for baseline (drift) correction. A Drift is a sample used for sensitivity drift correction. Usually a standard at ± 80% of the range is used. For the correction at least two cups with identical solution are required. A Standard (1..99) is a sample with known concentration. Based on a series of standards increasing in concentration a calibration curve is created against which the unknown samples are calculated. An Unknown is the actual sample being measured. The concentration is calculated based on the calibration curve and the baseline/drift correction. A Drift Ignore is a drift that is not included in the calculations for the sensitivity drift correction. The value is measured as an unknown.

69 Chapter 4 Tables 4-3 WI A Wash Ignore is a wash that is not included in the calculations for the baseline drift correction. The value is measured as an unknown. ICV An Initial Calibration Verification is used when CLP is applied. It is an independent standard at a concentration other than that used for instrument calibration, but within the calibration range (for more information see also chapter 5.3.5). CCV A Continuing Calibration Verification is a standard in the mid-range of the calibration curve, but at a different concentration than the ICV and the CCV and may not be used as a standard in the calibration curve. It is used to ensure calibration accuracy during an analysis run. ICB The Initial Calibration Blank is a blank sample prepared with reagents and reagent water and in the same concentration as used in the analytical sample preparation. This ICB is measured immediately after the ICV. CCB A Continuing Calibration Blank is prepared with reagents and reagent water and in the same concentration as used in the analytical sample preparation. It shall be analyzed immediately after the CCV. PBS A Preparation Blank Sample is used to monitor for possible contamination. The PBS contains all the reagents and in the same volume as used in pretreatment of the samples. LCS A Laboratory Control Sample is a sample of known composition. Laboratory Control Samples are analyzed using the same sample preparation, reagents, and analytical methods employed for the samples. LFB A Laboratory Fortified Blank is made by reagent water or other blank matrices to which known quantities of the method analytes are added in the laboratory. SR A Spike Recovery is a sample that is prepared by adding a known quantity of the analyte to a prepared sample. The SR is designed to provide information about the effect of the sample matrix on the measurement methodology. DR Duplicate Recovery sample determinations are used to demonstrate acceptable method precision by the laboratory at the time of analysis. D1..20 A typed Drift (1..20) is a channel dependency drift that can be selected in the method. Q1..20 A series of Quality Control samples is used for quality control of the method. Samples with (different) known concentrations are used. B A Blank sample is prepared with reagents and reagent water and in the same concentration as used in the analytical sample preparation and is used to eliminate any matrix effect. The blank results are subtracted from the samples that follow the blank until the next blank in the table. B1..20 Blanks (1..20) are blanks for the different channels (B1 for channel 1, B5 for channel 5, etc.) SK A Spike is a sample that is automatically spiked with a predefined volume and concentration of a stock solution to the previous sample in the table. SK1..20 Spikes (1..20) are spikes related to a certain channel. I Ignore is used to ignore the result. For more information about the use of these sample types refer to other chapters in this manual.

70 FlowAccess V3 4-4 The Table Entry screen has three main menus: File, Edit and Tools and a toolbar with nine easy access buttons for the most used functions. When clicking in the table with the right mouse button, also a menu with frequently used options is displayed. The File menu has eight options: New Table, Save, Save as, Export, Import, Table Wizard, Print and Exit (see Figure 4.2). The option New Table is not available when the table is edited. All options can be accessed via a keyboard shortcut (Ctrl + letter). - New Table can be used to create a new table after saving a created table, without closing the screen When this new table is saved, this table is attached to the sampler. (only when created by means of the Figure 4.2 Table tool). - Save is used to save a new table or edited table under the same name. The option can also be accessed from the toolbar. - Save as is used to save a new table or to save the table with another name. A dialog box is displayed where the (new) name is entered and the type of table is selected. - Export is used to export the table to a text format file or an Excel file. For both options a dialog box is displayed where the (sub)directory can be chosen where the exported file is saved. - Import is used to import a table from a text format file or an Excel file. For both options a dialog box is displayed where the (sub)directory can be chosen from where the table to be imported can be opened. - Table Wizard is used for easy setup of a complete sample table (for more explanation see chapter 4.4). - Print is used to print the table. This option is also accessible from the toolbar. - Exit is used to close the screen to return to the Active system. The Edit menu has ten options: Cut, Copy, Paste, Search Clear Cells, Auto Number, Fill Dilution Location, Insert Row, Delete Row(s) and Repeats Info (see Figure 4.3). All options except Search can be opened by clicking on the right mouse button in the table. All options can also be accessed via keyboard shortcut Ctrl + Shift + letter. - Cut, Copy, Paste and Clear Cells are the normal Windows options for replacing, copying, pasting or removing the Figure 4.3 contents of a cell. - Search is used to find a certain identity in the table. - Auto Number is used for automatic numbering of a number of sample positions or Identities. Insert the first position number or identity

71 Chapter 4 Tables 4-5 number and then select the positions to be numbered including the first position. Then select Auto Number. This option can also be accessed by clicking the right mouse button in the table and from the toolbar. - Fill Dilution Location automatically enters the location of the prediluted samples. Fill in the pre-dilution factor (2, 5 or 10) and then select the column for the dilution location and select this option. The location for each sample to be pre-diluted is entered, starting with the last available position in the last tray. This option can also be accessed by clicking the right mouse button in the table and from the toolbar. - Insert Row and Delete Row(s) can be used when complete rows are selected. Select a row by clicking on a row number. Insert Row inserts an empty row in the table above the selected row. Delete Row(s) deletes the selected rows. Before rows are deleted the deletion always has to be confirmed. - Repeats Info displays a dialog box (see Figure 4.4) where the number of sample pickups for drift/wash, standards and samples is multiplied by the number as set in the dialog box. Note: In this box the number is a multiplication factor. When already a duplicate is present in the table and 2 is entered for the repeats, the table is extended to 4 pickups per sample. After re-opening the dialog box the factor is set back to 1. The Tools menu has three options: Navigation, Tray Selection and ColorCoding (see Figure 4.5). - Navigation is used to navigate Column wise or Row wise through the table. This option can also be accessed by clicking the right mouse button in the table. - Tray Selection is used to change the used racks on the sampler. Only the racks that match with the set-up configuration are available. - ColorCoding is used to display the color code for the samples as used in the Table Wizard. When this option is deselected, the table is displayed without color coding. This option can also be accessed by clicking the right mouse button in the table. When the right mouse button is clicked in the table a menu is displayed (see Figure 4.6). The options in this menu are also accessible from the main menus in the screen. The toolbar displays nine easy access buttons for frequently used options in the main menus. Figure 4.4 Figure 4.5 Figure 4.6

72 FlowAccess V Table wizard Often, the practice of laboratories is to perform analysis runs of certain parameters according to set analysis routines. The number of analyses might change daily, but the course of the run basically stays the same. A routine run looks like this: An analysis starts with a tracer to start-up the system. This should be a fairly high standard as the program collects the data of the initial wash after the Start ignore time (defined in the Methods, see chapter 5.3.1). It waits for the first peak that exceeds the start value. The program needs this value to start the detection of the channel. After the tracer follows a wash ignore to bring the signal down to the baseline. These first two cups are not included in the calculations for the correction of the chemical and baseline drift. The calibration of the analysis is then performed. Standards, (usually) increasing in concentration build the calibration curve. A drift and a wash for the correction of chemical and baseline drift follow the standards*. The program searches for the second drift and wash. The number of positions between these drifts and washes is defined as the drift/wash interval. The course of the sample table is followed, automatically placing a drift and a wash after the preset interval. The run ends with a drift and a wash. *Correction for baseline drift and chemical (sensitivity) drift are necessary for correct calculation of the measured peaks. The baseline and the sensitivity can change due to differences in temperature or reagent changes. Each sample between two washes and drifts is first corrected for the baseline drift (wash-wash difference) and then for the sensitivity drift (drift-drift difference). For calculation formulas for baseline and chemical drift correction see chapter The table wizard defines the standard course of the analysis run, so the positions of the start-up samples, the standards, the drift-drift intervals and the wash-wash intervals, as well as the number of samples are placed in the wizard. The pre-dilution factors (manual and automatic), the weight factor and the identification of the unknowns are not put in, they are put in later when the table has been created. Note: A table for a SA1000 sampler cannot be setup with the table wizard as this sampler has no special positions for standards, drifts and washes. Also a table with only samples is not setup with the table wizard as the wizard requires all items filled in. These tables are created directly in the Table Entry screen (see chapter 4.6). 4.5 Creating a table with the Table Wizard Creating a table by means of the Table menu 1. Click on the Table menu and select Create Table. The New table screen is displayed (see Figure 4.7). Figure 4.7

73 Chapter 4 Tables Select the sampler for which the table is to be created and click OK. The Select trays screen is opened (see Figure 4.8). Figure Check the trays that are present on the sampler (and uncheck trays that are not present, make sure to control the complete list, refer also to the table in Chapter 3 of the sampler manual,) and click OK. The Table Entry screen is opened (see Figure 4.9). The layout is dependent on the selections made in the system setup (see Preferences, chapter ). Figure 4.9 Note: If the selectable trays do not match the ones present on the sampler, check your system setup. Either your sampler configuration was not setup properly, or the wrong racks are present. 4. Select the Table Wizard tool. The Table Wizard screen opens (see Figure 4.10) with the trays displayed as selected in Figure 4.8. The left part of the screen displays all items required for the table setup. For each item a screen will be displayed where the items are set.

74 FlowAccess V3 4-8 Note: When the Table Wizard is used only a complete table with standards and drifts can be created, as the Table wizard needs the settings for all items. If a sample table with only samples is required, the rows can be deleted later in the Table Entry screen. Figure Select Tracer or click Next to insert de settings for the tracer (see Figure 4.11). 6. Select the cup position of the tracer and select the wash option if required. 7. If a Drift ignore and Wash ignore are to be placed in the table, enter the position of the drift and wash. Figure Select Drift/Wash or click Next to insert the settings for the Drift/Wash (see Figure 4.12). Figure 4.12

75 Chapter 4 Tables Enter here the Drift and Wash position for the sensitivity and baseline correction and the interval for this correction. More than one position can be entered for the drift. These positions can be overruled by the settings in the sampler (see chapter ) when the sampler is saved. Make sure to put drift liquid in the set positions! Select the number of pickups for drift and wash for each interval. 10. Select Standards or click Next to insert the information about the standards (see Figure 4.13). Figure Enter the lowest used standard as set in the method and its position on the sampler. Enter the total number of standards. Enter the number of pickups for each standard of the calibration. 12. Select Samples or click Next to insert the information about the samples (see Figure 4.14). Figure Enter the total number of unknown samples and the start position of the samples. Check the result selection and if the samples are to be post-diluted (only if a diluter is present on the sampler). Enter the number of pickups for each (duplicate, triplicate, etc.) for each sample of the table. Note: The multiple pickup for drifts, washes, standards and samples can also be added by means of the option Repeats info after creation of the table. This box displays a multiplication factor. However, when this option is used, the drift/wash interval is not adapted and ahd had to be changed manually.

76 FlowAccess V If CLP has to be applied select CLP Setup or click Next to set the positions for the CLP samples (see Figure 4.15). Check the tick box and enter the positions of the CLP samples on the sampler. Figure Select Other Setup or click Next if spike recovery samples or duplicate recovery samples are present in the table (see Figure 4.16). Figure Enter the positions and the interval. Figure 4.17

77 Chapter 4 Tables Click Next and then Create. The screen is closed and the table is displayed with the settings of the Table Wizard (see Figure 4.17 for an example). Note: Automatically a drift/wash is placed after the Tracer in the table, but the calibration is usually not drift/wash corrected. This drift/wash can easily be removed from the table by selecting these two rows and clicking the right mouse button and selecting Delete Rows. 18. Enter the identification for the Unknowns and, if required, the weight, the external dilution factor and pre-dilution factor (see chapter 5.8). 19. Press Save to save the table, enter a name for the table (the table name has to start with a letter) and press OK. If the drift positions in the table do not match the positions as set in the sampler settings a pop-up message is displayed if you want to update the drift cup details to the positions as set in the sampler settings. If Yes is selected the table is saved the corrected drift positions. 20. The table is not yet loaded on the sampler. 21. To load the table: Select the Table tool and then click on the sampler. The Load Table screen is displayed (as in Figure 4.19). 22. Select Load an existing Table and click OK. The table is now loaded on the sampler. A table with CLP samples may look as displayed in Figure Figure 4.18 Warning: Do not run the same sample table on more than one sampler at any time; a conflict may occur. Even if two samplers of the same type are present in the system, make sure to separate the tables for each sampler by their names.

78 FlowAccess V Creating a table with the Table tool 1. Select the Table tool and then click on the sampler for which the table is to be created. The Load Table screen is displayed (see Figure 4.19). 2. Select Create new Table and click OK. The Select Trays screen as in Figure 4.8 is displayed, but only if no table exists for this sampler. 3. Proceed with step 3 till 19 of chapter to create the table. The table is automatically loaded on the sampler. 4.6 Creating a table in the Table Entry Figure 4.19 screen A table can also be created directly in the Table Entry screen. Especially tables with only samples or a table for the SA1000 are created here. The table can either be created by means of the Table menu or the Table tool. In this way also sample tables without standards and drift/wash interval can be created. Procedure: 1. Follow either chapter step 1 and 2 or chapter step 1 and Select the trays that are used on the selected sampler and click OK. The Table Entry screen is displayed. 3. Select the first cell in the Type column. When the cell is selected a down arrow is visible. 4. Click on the down arrow to display a list of options for the type of sample. 5. Select T (tracer). The T is entered for the type. When a type is entered always automatically an identity is added. When a tracer or a drift is entered automatically a wash is entered on the next row. (If you do not want a wash following the tracer, click on the row number before the row to select the row, right click to open the menu and select Delete Row(s) ). Note: As the first peak has to be detectable by the program, do not place a wash as the first sample in the table, but always a high standard. 6. Go to the next row. Select S1 for the type. 7. Select S2, S3 S4 and S5 in the same way. Note: Skip step 6 and 7 if a table with only samples is to be created. 8. Go to the next row and select D. A wash (W) is automatically added on the next row. 9. Go to the next row. Select U for the type. The identity Unknown is added. This identity can always be replaced by another identity. The selection automatically jumps to the next row. You can also type U and press Enter. 10. Select as many U type samples as there are samples to be analyzed. 11. Finally enter a D to enter a drift. A wash is automatically added on the next row.

79 Chapter 4 Tables When drift/wash intervals are required, select the position where you want to insert a drift/wash and right click to open the menu and select Insert row. Do this once more and select then D for the type on the first inserted row. Do this for all required intervals. 13. Go back to the beginning of the table. 14. Select the Position cell for the tracer and type the position of the tracer in the sampler. In this case ST5 (for SA1050, SA1070, SA1074, SA1075 and SA1100 with standard positions). Note: For SA1000 and SA1100 with the tray with 100 sample positions the standard positions have no special positions and are placed in normal sample positions (1 to 80 respectively 1 to 100). For SA1000 also the wash position is a normal position. However, for the SA1100 Wt must be selected for the wash position. 15. Do the same for the wash. Usually Wt, which is the position of the rinsing vessel (not for SA1000!). 16. Type the position for the drift. In this case ST4 (except for SA1000). 17. Type the wash position (only for SA1000 a normal cup position!). 18. Type the position for the first standard (ST1) (except for SA1000). 19. Select this cell and all position cells for the standards. Click Auto Number on the toolbar or right click and select it from the menu. All selected position cells are auto-numbered with their tray racks. Note: For a SA1000 sampler the Autonumber function can be used for the complete table. Only type the first position, then select the cell and all other positions of the table and click Autonumber. 20. Type the position for the drift and the wash. 21. Type the first unknown sample position. Select this cell and all the position cells of the unknowns until the next Drift. Click Auto Number on the toolbar or right click and select it from the menu. All selected position cells are auto-numbered. 22. Repeat step 20 and 21 until all positions are entered. 23. Now a specific identity for the samples can be inserted. Select the Unknown cell and type the identity. If the samples have an increasing number the Autonumber function can be used. The table may look like displayed in Figure When multiple pickups are required this can be achieved by means of the repeats info option in the Edit menu (see chapter 4.3). 25. When no automatic pre-dilution is used, save the table by clicking the Save button on the toolbar. Type a name for the table in the displayed dialog box and click OK. 26. Click Exit to close the table. When the table is opened again the name of the table is displayed in the title bar. 27. Note that when a position is entered in the table, automatically 1 is entered for external dilution factor, weight and dilution factor. If necessary, these factors can be changed to the real factors. Warning: Never try to edit an existing table for another type of tray. Always remove the table and create a new table after selecting the required tray in the Tray selection screen. Note: CLP samples can be inserted as desired. For inserting spiked samples see chapter 4.9.

80 FlowAccess V Figure 4.20 Note: Remember that this is FlowAccessV3 TM when copying and pasting rows. Use the right mouse button or the Copy option from the Edit menu, as CTRL+C and CTRL+V won t work. Select the same fields for pasting as were selected for copying, as otherwise not all channels are pasted.

81 Chapter 4 Tables Creating a table for 2 or more needles When 2 or more needles are present on the sampler (SA1050 and SA1070 max. 2 needles, SA1074 and SA1075 max. 4 needles), dependent on the sampler type and number of needles, different tray types are available for the samplers. Only half or less positions are available for each needle for the standards and samples. The table can be setup with the table wizard. The wizard creates the table for needle 1 and copies this table to the table for the other needle(s). When the table is created, the table shows tabs with the needle numbers at the bottom of the table (see Figure 4.21). The active tab is underlined with an orange color (in Figure 4.21 the table for needle 2 is active). When the table is created the identification in the other needle table(s) can be changed, but not the positions as these are fixed by the position in the table for needle 1. Figure 4.21 When samples are added to the table for Needle 1 only and the table is saved, the following message is displayed (see Figure 4.22): Figure 4.22 When Yes is selected an Ignore is placed for the other needles and the results for the other needles are not calculated.

82 FlowAccess V Creating a table with automatic pre-dilutions When some or all unknown samples are to be pre-diluted (SA1070, SA1074, SA1075 and SA1050), the automatic pre-dilution factor has to be entered under Dilution factor. Also the location for the diluted sample has to be entered. If the columns for dilution factor and dilution location are not visible in the screen, the diluter is not selected in the Hardware setup. Note: Under External DilFactor only the manual pre-dilution factor has to be entered. Always insert this factor after the position is entered. Procedure: 1. Select the cell in the column Dilution factor for the sample to be prediluted. 2. Click on the down and select the dilution factor (2 to 50) from the list. Note: For pre-dilutions > 50 the pre-dilution utility of the sampler can be used. Refer to chapter Do the same for all samples to be pre-diluted. 4. Select the column Pre-dilution position (including the heading) and click on the icon Auto Fill Dilution Location in the toolbar (or select it from the Options menu). Automatically the dilution locations are filled in, starting from the last available position in the selected sample racks (for example D35 for SA1050, see Figure 4.23). Note: When no dilution locations are entered, a message is displayed when you want to close the screen that locations have to be entered. Figure 4.23 Table with automatic pre-dilutions Note: When a pre-diluted sample needs to be mixed, this is set in the Group (Edit group).

83 Chapter 4 Tables Creating a table with spikes Spiked samples (type SK) can be put in the table, but only when a diluter is present on the sampler, as the sample will be prepared from a stock and the sample previous to the spiked sample. Make sure to place an empty tube on that position. When the table is saved, first a message is displayed that the table is saved successfully. Then a screen is displayed where the information has to be entered about the spike (position, concentration and volume) and the volume of sample to be taken from the previous sample (see Figure 4.24). Press OK to save these settings. Figure 4.24 Table with spiked sample 4.10 Importing tables A table made in text format and Excel format can be imported in FlowAccessV3 TM. A text format table can have different formats. As soon as a table in text format is imported a screen is displayed where the required layout can be defined. The text file may be a complete sample table or a table with only unknowns, which is later combined with standards to a complete sample table. To prepare a text format table: 1. Open Wordpad or Notepad from the desktop. 2. Type the position of the first sample in your table. 3. Enter a delimiter and then enter the type of the sample. 4. Enter a delimiter and enter the identity of the sample. 5. If more relevant information is to be entered, these can be entered in the same way as described in step 3 and If a diluter is present, you can enter the pre-dilution factor here or later in the table when it is imported. 7. Press ENTER.

84 FlowAccess V Repeat step 3-7 until all samples are inserted. Your file may now look as in Figure Here the position, type, identity and pre-dilution factor are defined. 9. Select Save from the File menu and type a name for the table in the Save as dialog box. Save the table as.txt file. Figure 4.25 Example of text file 10. Select the directory where the file is to be stored and click Save. Import of text format files: 1. Click the Table icon and then on the sampler. 2. Select Create new table and click OK. 3. The Table Entry screen is opened with an empty table. 4. Select Import from the File menu and select the type of file (text format) to be imported. Figure 4.26

85 Chapter 4 Tables Move in the dialog box to the directory where the file is stored, select the file to be imported and click Open. The Delimiter Settings screen is displayed (see Figure 4.26). 6. Select the delimiter used in the text file. 7. Enter the start row and the end row to be imported and press Load Column. 8. (De)select the modules from the list. 9. Click in the column Select columns behind Position. A drop-down menu displays the number of columns of the text file. 10. Select the column number from the text file to be used for the Position. Do the same for the other items of the text file and click OK. The file is imported and displayed in the table (see Figure 4.27). Figure 4.27 Imported file 11. A message is displayed that the file is imported successfully. Click OK to close this box. 12. Select Save as from the File menu and type a name for the table. To prepare a table in Excel format: 1. There are two ways to create an Excel table: Either open Excel and create a table with the same columns as in the table in FlowAccess V3 or create a table with only the relevant columns. In both cases type the headings for the columns in row 1 of the Excel file. 2. Rename the tab from Sheet1 to Needle1. In the case a table for two or more needles is created, the tabs for these needles have to be renamed also. 3. Save the table as.xls file. (See Figure 4.28).

86 FlowAccess V Figure 4.28 Excel table Import of Excel format files: 1. Click the Table icon and then on the sampler. 2. Select Create new table and click OK. 3. The Table Entry screen is opened with an empty table. 4. Select Import from the File menu and select the type of file (excel format) to be imported. Move in the dialog box to the directory where the file is stored, select the file to be imported and click Open. The Import Excel Options screen is displayed (see Figure 4.29). Figure A. In the case that the Excel file was created with exact the same columns as in the table select the Simple mode. The file is directly imported in the table. When the Excel file was created with a different format, then select Advanced mode. In this case the Delimiter Settings screen as displayed in Figure 4.26 is displayed. 6. Enter the start row and the end row and then click Load Column. 7. Select the column number from the Excel file to be used for the Position. Do the same for the other items of the text file and click OK. The file is imported and displayed in the table (see Figure 4.27).

87 Chapter 4 Tables Exporting tables 1. To export a table from the Table Entry screen: 2. Select Export from the File menu and select the export format. The Save as dialog box is displayed. 3. Type a name for the table to be exported. 4. Select the directory where the table is to be saved and click Save Edit table A. Click on the Table icon and then click on the sampler. Select Edit Table. The table attached to the system is displayed in the Table Entry screen (for an example of a table see Figure 4.20). Edit the table according your wishes. For instructions see chapter 5.5 the Table Entry screen. Save the changes in the table and close the screen to return to the Active System screen. B. Select Edit Table from the Table menu and click Open. Edit the table according your wishes and save the changes. Close the screen to return to the Active System screen. Note: A sample table cannot be edited in making it temporary shorter by inserting and E (=end) as Type in the table. When an existing table is to be made shorter, all positions below the last sample in the new table have to be deleted. If you want to keep the longer table, store the edited table with a new name Remove Table Select Remove Table. The table attached to the displayed sampler is removed. The table is not deleted from the list of tables. A table can only be deleted from the list in the Edit Table screen from the Table menu. Note: Delete old tables from the file list if they are not used anymore. This is to prevent unnecessary growing of the database.

88

89 FlowAccess V3 CHAPTER 5: METHODS Preface: This chapter gives detailed instructions about all aspects of the methods on the system

90

91 FlowAccess Chapter 5 Methods METHODS Methods have to be defined for the chemistries on the analyzer. These methods contain the required information for the software for calculation of the results of the analysis run. There are two ways to define a method: 1. With the Methods menu, methods can be created, edited and deleted. 2. When clicking the Methods icon on the toolbar (the icon is added to the pointer) and then on a chemistry in the analyzer, a method attached to that chemistry can be created, edited or removed or an existing method loaded (attached). 5.1 Methods menu - New Method is used to create a new method. The New Method screen (see Figure 5.3) is displayed, where the method can be defined. This method does not show a chemistry name, only the module where the method will be attached. When the method is saved a name has to be given and the method is attached to the displayed module. In the same screen other methods can be defined after selecting the required module. - Edit Method is used to edit or delete a method. A dialog box called Edit and Delete Methods with the existing defined methods for the current displayed system is opened (see Figure 5.1). Also methods that were defined for another system can be displayed by selecting Show all methods. However then the displayed methods cannot be edited, only deleted. - Custom method is used Figure 5.1 Edit Method to display defined methods that are not assigned to a module. New methods can be defined and existing methods edited and deleted. For instructions to create or to edit a method see chapter 5.3 and 5.5.

92 FlowAccess V Methods tool Click the Methods icon and then click on the module where the method is attached or to be attached. A dialog box is displayed with four options (see Figure 5.2) of which at least two options are always gray. When no method is defined the last three options are gray. When no method is attached to the channel, but methods are defined both last options are gray. When a method is attached to the channel both first options are gray. An attached method has to be removed before another (new or existing) method can be attached to the channel. 5.3 Create New Method Figure 5.2 Load Method Select Create new Method in Figure 5.2 or select New Method from the Methods menu. The New Method screen is displayed (see Figure 5.3) with six tabs: Peak Detection parameters, Peak Detection type, Calibration, Result, QC and CLP Concentration and Post Dilution. In each tab specific settings for the method are set. The screen displays one main menu: File. Also a toolbar with two easy access buttons Save and Exit is available. Figure 5.3 Method setup File displays two options: Save and Exit. - Save saves the edited method or opens the Save Method screen where a name has to be typed for the defined method. As soon as a name is typed for the method the name is displayed in the title bar of the screen and as Method File Name.

93 FlowAccess Chapter 5 Methods Exit is used to close the screen. The left pane of the screen displays the available modules of the system. The upper part of the right pane displays the system, the module name where the method will be attached and the method name (only if the method was edited). Save the method when all necessary information about the method is set in each tab. Click Exit to return to the Active System or select the next method to be defined in the left pane Peak Detection parameters In this tab all parameters of the peak detection are defined (see Figure 5.4). The screen displays the default settings. These defaults can be changed if required for the application. Figure 5.4 Peak Detection parameters Detect peak based on defines how the peak is detected. This is currently always on peak height. Rawdata Unit defines the units that are displayed in the real time screen. A selection between Absorbance units and Digital units can be made. Start Value Tracer defines the increasing and falling value in digital units or absorbance units of the first peak. At the end of the start ignore time, after collecting the data for the initial wash the program waits for the moment the ascending and falling slope of the first peak exceeds the start value. The program also measures the time between the ascending and falling slope of the peak. This time must exceed 30% (= default peak width) of the total sampling time (sample time + wash time + air time). The program then knows that the tracer or the first peak has arrived. From here the program predicts the arrival of the next peak based on the sample time. Default setting is 1,000 DU. This is only for a 16-bit detector (8502 or 8503 interface). The value should be adapted to at least 50% of the value of the highest standard, which is dependent on the application. For a 32-bit detector (8503 or 8505) the setting should be 100 x higher (1 AU = 1,000,000 DU), otherwise the program is triggered even at noise level.

94 FlowAccess V3 5-4 Peak Width Tracer defines the percentage of the total sampling time that the peak needs to be, in order to be recognized as the tracer (see also Start Value Tracer ). Default Setting is 30%, but should be changed into a lower value in the case of narrow peaks. Height Reject defines the minimum value in digital units or absorbance units a peak (except the tracer and the wash) is expected to attain (both rise and fall) to be detected as a peak. All values lower than this value within this measure window will be marked as absent. The next expected sample peak will be measured in the same measure window interval and no time shift correction will take place. Default setting is 300 DU (this is for a 16-bit detector). The default value should be adapted to at least 2% of the value of the highest standard, which is dependent on the application. For a 32-bit detector the values must be set 100 x higher. Start Ignore time defines the time the program has to wait in seconds before the data of the Initial Wash of that method has to be collected. It is advisable to enter the time it takes for the first sample to reach the flow cell minus 30 seconds. This time may be different for each method and is measured by a stopwatch. The default setting is 120 seconds. End Ignore time defines the time the program for that method has to wait before the data of the final wash, at the end of the run, is collected. In general it is advisable to enter a time equal to the Sample Time + Wash Time. The default setting is 120 seconds. Trigger Time defines the time limit that a channel is started in the case the Start Value might not be reached (e.g. in a blank channel). The timer starts counting the seconds after the Start Ignore time has passed. If the analysis run was indeed started up by exceeding the trigger limit the first sample in the results table is flagged with an L. The default setting is 900 seconds. This trigger time takes care that in case of a failure in a chemistry, the channel is still started and that the analysis is closed and a run file is created. The data of the other channels is ok. Measure Window counts a measure window around the next predicted peak. The program checks if the peak top is in the middle of the window. If it is not in the middle, the next window will be corrected accordingly. In case the rising and falling slope of the peak doesn t exceed the Height Reject, no time correction takes place. When only a down going or up going slope is detected in the window, the middle of the window is marked. This is also the case when first a down going slope and then an up going slope is detected in the window. In case of a wash sample the lowest point in the window is marked. The Measure Window is a certain percentage of the total sample cycle (sample time + wash time + air time). The default setting is 75%. By default there is no visual display of the measure window itself (can be selected in the Realtime window). Peaks that arrive too late (L) or too early (E) in the measure window, multiple peaks, missing peaks (A) and normal peaks (N) will be flagged in Real Time and in the Results. Over range results (O) will be flagged in the results (when this column is selected) and in the report (when this column is selected). Positive Wash. Select this option when the method is used as a blank channel. The detection method normally searches for the lowest point in case of a wash sample. Under certain conditions the blank sample produces a peak, which is then wrongly detected. When Positive Wash is selected the detection method looks for the highest point, thus avoiding wrong detection.

95 FlowAccess Chapter 5 Methods 5-5 Reaction Type selects how the chemistry is detected. Three reaction types are available: Colorization (color increase), Decolorization (color decrease) and Colorization/Decolorization. Colorization/Decolorization is used for measurement of peaks on detectors like density meters. Note: If the Reaction Type is set as 'Decolorization', then the new raw data is calculated as Max DU - received raw data value. For non 32- bit it is received raw data value. For 32-bit it is (2 31 )-1 - received raw data value Peak Detection type Five calculation types for the peak detection are available: Min/Max peak detection, Chebyshev Approximation, Least square method, Min. Standard Deviation method (with optional Spike detection peaks) and Savitsky-Gohlay filtering method (see Figure 5.5). Figure 5.5 Peak Detection type For optimum peak detection the chemistry can be detected and calculated by several calculation methods. If more than one calculation method is selected, the result is calculated by the mean of these calculation methods. When more detection methods are used, the chance that the result was calculated correctly increases, especially when the chemistry channel does not run smoothly. The results calculated by the selected peak detection methods will be compared to each other. If the result of one detection method is significantly different from the other detection methods (larger than the set tolerance), this detection method (for this channel and this peak only) is skipped, and the peak calculation is done with only the remaining values and a warning (flag) is given in the presentation of the result.

96 FlowAccess V3 5-6 If the result of more than one detection method is significantly different from the other detection methods (larger than the set tolerance), these detection methods (for this channel and this peak only) are skipped, and the peak calculation is done with only the remaining values and an error (flag) is given in the presentation of the result. For peak detection of normal peaks the methods A+B+C+D+E can be selected. Method A is obligated for peak detection in real time. When the peaks are wide and flat preferably use methods A+D+E.. A. Min/Max peak detection method This detection type is standard detection method. Max. peak detection for colorization analysis and Min. peak detection for de-colorization analysis. This detection method is obligated for peak detection in real time. It can be deselected in PostAnalysis. B. Chebyshev Approximation method In this method a 5 th order polynomial is built from the peak data, which is differentiated where after is looked for min. and max. values. This peak detection method generates very good results, except for flat, wide peaks 1,2. C. Least square method In this method a 5 th order polynomial is built from the peak data with the least square method, which is differentiated where after is looked for min. and max. values. This peak detection method generates very good results, except for flat, wide peaks 2,3. D. Minimum standard deviation method In this method a floating mean is calculated on the peak and the peak max. is calculated with the minimum standard deviation of the mean. This method is suitable for all kind of peaks, but is more sensitive for interferences than the previous mentioned polynomial approximations. If the chemistry shows a lot of spikes this detection method should be selected. Here the Spike detection peaks can be selected, wherefore the Spike threshold and the Spike determination range must be defined Spike Threshold selects the time in seconds until the peak is considered to be a spiked peak and not a peak to be measured. The threshold can be set between 2 and 10 seconds. Spike Determination Range sets the time in seconds that the software has to wait after a spike is measured for the measurement of the real peak. E. Savitzky-Gohlay filtering method This methods is analog to method D, however the peak is first filtered with a Savitzky-Gohlay 2 filter. The filter factor is References: 1. Shan S Kuo, Numerical Methods and Computers, Addison Wesley, Reading, Massachusetts 1965, Library of Congress Catalog Card No , p.228ff. 2. Numerical Recipes in C The Art of Scientific Computing-, Cambridge University Press, Library of Congress Catalog Card No , ISBN , 3. Alan R Miller, Fortran for Scientists and Engineers, Sybex Verlag, Library of Congress Catalog Card No , ISBN , p99ff

97 FlowAccess Chapter 5 Methods 5-7 Peak Detection Tolerance The peak detection tolerance is the max. allowed difference for the peak detection methods. It is advised to set the value on 2.5 % of the Tracer peak height or height of the highest standard. As the height of the Tracer is not known in advance the value is set by default on Absorption units. Gain Factor to decrease the baseline (0.3 Absorption Units per digit) Damping options This option can be used for damping of the raw results acc. Savitzky - Gohlay. Default is set on No Dampingr. For damping eight different options are available. The set damping is dependent on the stability of the system and the range of the method. The optimum damping option for a method (if required) is usually established at Skalar. The filter can be changed in PostAnalysis (in Data valuation, chapter and Peak picking parameters, chapter 9.3.2) Calibration Here all aspects of the calibration is set (see Figure 5.6). Calibrate is selected when performing the calibration for normal analysis. If the method is a blank, the option is deselected. Figure 5.6 Calibration Calibration Standards. Type the concentration for the standards used for the calibration in the right pane; automatically it is selected for the calculation of the calibration curve. During the analysis it is possible to deselect a standard for the calculation if the value for the standard seems to be incorrect (see chapter ) Calibration Type selects the calculation type for the calibration curve. Eight types of calculation are selectable: 1 st Order according ISO nd Order according ISO rd Order 1 st Order forced through zero

98 FlowAccess V nd Order forced through zero 1 st Order Inverse Logarithm 2 nd Order Inverse Logarithm AutoCalibration In the first order the most possible straight line is calculated using linear regression. The second order calibration curve is calculated using parabolic regression. The inverse logarithm calculation is built by means of an exponential function (see also chapter for the used formulas) Concentration Units. Here the dimension of the concentration is entered. This dimension is also displayed in the heading of the results table and in the reports. Standard name. Type the name of the used standard for the calibration. Calibration from another run It is possible to calculate the results of an analysis run with the aid of the calibration of a previous run. This is only possible when Calibrate is deselected. Choose a run file that also contains the same method in the same range. This can be done before, during or after the run. It is also possible to change a previous selected calibration into another calibration Results Here the information for the display and calculation of the results is set (see Figure 5.7). Figure 5.7 Results No. of Decimal places Enter the number of digits after the decimal that must be displayed in the results (default is 2) Perform Wash Correction Select this option to apply baseline correction. See also chapter for explanation Perform Drift Correction. Select this option to apply sensitivity drift correction. See also chapter for explanation

99 FlowAccess Chapter 5 Methods 5-9 Drift Rejection Percent Enter the max. percentage a drift is allowed to deviate (+or-) from the previous drift before it is rejected for the calculation. In case the % is exceeded, the drift will be skipped and the batch of samples will be corrected by the previous drift if this is within limits. Low Drift refuse When this option is selected the mean of all measured drifts in the analysis run is used. This mean is calculated newly after any next drift measured. If the value of the last measured drift is exceeding the preset %, the drift will be skipped. The value will not be used to calculate the mean of all drifts. The samples in between the drifts will be corrected with use of the mean. Low drift refuse value Enter the maximum deviation in % a drift may differ (+ of -) from the mean of the drifts measured before it is rejected for the calculation. Alarm Upper Enter the upper limit of the results of the unknown samples. If the alarm is exceeded the results will be flagged Alarm Lower Enter the lower limit of the results of the unknown samples. If the alarm is exceeded the results will be flagged Note: To obtain result flags the value of both upper and lower level must be entered and the value of the lower level must be > QC & CLP concentration This tab defines the concentration of the QC samples and their alarm limits, as well as the concentration limits for several types of CLP samples (see Figure 5.8). CLP stands for Contract Laboratory Program. Figure 5.8 QC The QC limits can be set can be set as an absolute value or as a percentage of the concentration. For each type of CLP sample the upper and lower limit can be set as required. When the CLP is selected the value type can be set on absolute or relative. Then the value of the calibration type (either r square or V xo ) must be entered.

100 FlowAccess V The following CLP samples can be set: - Initial Calibration Verification (ICV) For this ICV an independent standard is used, at a concentration other than that used for instrument calibration, but within the calibration range. An independent standard is defined as a standard composed of the analyte from a different source than those used in the standards for the calibration. The ICV is prepared in the same matrix as the calibration standards. The value should be within the control limits of 85% and 115%. - Initial Calibration Blank (ICB) The ICB (and CCB) is used in establishing the calibration curve. The ICB shall be analyzed immediately after the ICV at a frequency of 10% or every 2 hours during the run, whichever is more frequent. The ICB is prepared with reagents and reagent water and in the same concentration as used in the analytical sample preparation. The absolute value of the ICB result may not exceed the CRQL*. - Continuing Calibration Verification (CCV) The CCV is used to ensure calibration accuracy during each analysis run. For this CCV a standard in the mid-range of the calibration curve is used, but at a different concentration than the ICV and the CCV may not be used as a standard in the calibration curve. This CCV should be analyzed and reported at a frequency of 10% or every 2 hours during the analysis run, whichever is more frequent and also at the beginning of the run and after the last analytical sample. The value should be within the control limits of 85% and 115%. - Continuing Calibration Blank (CCB) The CCB shall be analyzed immediately after the CCV at a frequency of 10% or every 2 hours during the run, whichever is more frequent and also at the beginning of the run and after the last analytical sample. The CCB is prepared with reagents and reagent water and in the same concentration as used in the analytical sample preparation. The absolute value of the CCB result may not exceed the CRQL*. - Laboratory Control Sample (LCS) A Laboratory Control sample is a sample of known composition. Laboratory Control Samples are analyzed using the same sample preparation, reagents, and analytical methods employed for the samples. The result must be within 80 and 120%. - Spike Recovery (SR) The Spike Sample Analysis is designed to provide information about the effect of the sample matrix on the measurement methodology. A spike sample is prepared by adding a known quantity of the analyte to the prepared sample. At least one spike sample analysis shall be performed on each group of samples of a similar matrix type or for each sample delivery group. The spike recovery must be within 75 and 125%. - Duplicate Recovery (DR) Duplicate sample determinations are used to demonstrate acceptable method precision by the laboratory at the time of analysis. Duplicate analyses are also performed to generate data in order to determine the long-term precision of the analytical method on various matrices.

101 FlowAccess Chapter 5 Methods 5-11 The duplicate sample is designed to provide information about the method precision by the laboratory at the time of the analysis. At least one duplicate sample analysis shall be performed on each group of samples of a similar matrix type or for each sample delivery group. The duplicate recovery must be within 80 and 120%, when the original and duplicate sample values are greater than or equal to 5x the CRQL*. *CRQL stands for Contract Required Quantitation Limit. This is the minimum level of quantitation acceptable under the contract statement of work. Also CRDL is used (Contract Required Detection Limit). During the analysis, the results for the CLP samples are checked against the limits and if the results are found to be outside the limits the action as set in the Group (see chapter 6.3.6) is performed. This can be either rerun, abortion of the run or continuing and flagging. If both upper and lower limit are set on 0 (zero) for a type of CLP sample, this sample will be ignored during the analysis, i.e. the results for these cups will not be checked against the limits Post Dilution In this tab the Post dilution criteria are set (see Figure 5.9). Figure 5.9 Post Dilution When a diluter is present on the sampler the samples can be automatically postdiluted when over range. Post Dilution Select the post-dilution option when automatic post-dilution on over range samples is to be applied. (Also select post-dilution in the group parameters, see chapter 6.3.5). Note: The Post-dilution option can also be set here for systems without diluter. In the Group settings the post-dilution is then not selected. When the threshold type HT>HI. STD is selected, the results that are calculated as Over-range are flagged with O in the results flagging column.

102 FlowAccess V Threshold type Select the threshold type to apply post-dilution. A choice can be made between: (Calculated) concentration, Corrected peak height (absolute height without baseline) Peak height higher than the highest standard. For correct calculation the sample should always be diluted when a sample result does not lie between the lowest and highest standard of the calibration curve. When a second order calculation curve is used, it might be possible that the curve is eventually parabolic and two results can be obtained. Then it is possible that a peak that is higher than the peak for the highest standard is not diluted as the result is calculated as lower than the highest standard. So for a second order calibration select as threshold type = peak height higher than the highest standard. Note: The threshold type Height higher than the highest standard cannot be used when the calibration of another run is used as this information is not available for the current run. Dilution Threshold Enter the concentration threshold or the height threshold for post-dilution. Post Dilution Factor Enter the required post dilution factor for this method. This factor will be overruled by the post dilution settings in the group settings, when different. Use Diluter/Use Diluter Valve Select Use Diluter. (Use Diluter Valve is for future use) 5.4 Load an existing method Defined methods can be attached to a channel by means of the option Load an existing method. When this option is selected the Select Method dialog box is displayed (see Figure 5.10) with all defined methods (also those created in Custom methods ). Select the method to be loaded and click Load. Figure Edit a Method Each method that is attached to a channel can be edited by this option. When this option is selected the Method screen of the selected method is displayed. The method can be changed as desired. Save the changes.

103 FlowAccess Chapter 5 Methods Custom method It is possible to create and edit methods that are not automatically attached to a channel. This may be convenient if a channel is used for analyses in different ranges. When this option is selected the Unassigned Methods dialog box is displayed (see Figure 5.11). Here new Methods can be created as explained in chapter 5.3, but the method is not attached to a channel. The methods can be edited and deleted from this screen. Figure Remove a Method A method that is attached to a channel can be removed. The method is not deleted from the list of methods by this action. A method can only be deleted by means of the Method menu: Edit Method or Custom Method.

104

105 FlowAccess V3 CHAPTER 6: GROUPS Preface: This chapter gives detailed instructions about grouping the methods for the analysis run.

106

107 FlowAccess Chapter 6 Groups GROUPS The methods to be analyzed in one run are grouped as they share one table. It is possible to create more than one group on a system with one needle on the sampler, but they will run on the same table. Only when the sampler is equipped with two (or more) needles, more tables are run as each needle runs its own table. A method can be selected only for one group at the same time. If you want to change the composition of a group, first the existing group has to be removed from the system and then a new group defined. Parameters as Post-dilution and Working standards are also defined here as a part of the group. 6.1 Create New Group 1. Select the Group button on the toolbar. The Group icon is added to the pointer. 2. Click on the sampler. All channels in the system are colored white. 3. Click on the channels to be grouped (a selected channel turns green) and then click with the right mouse button on one of the selected channels. A pop-up menu is displayed with one active option: Add Groups (see Figure 6.1). Figure 6.1 Add groups

108 FlowAccess V Select this option. A dialog box is displayed. 5. Type a name for the group. 6. Click OK. The box is closed and the name of the group is displayed on the right side of the screen with the same color as the channels of the group. 7. If desired click again on the sampler to select a new group. Select the channels and right click with the mouse to give the group a name (a channel that is already selected for another group cannot be selected). This group gets another color. As soon as a group has been created, three options are available from the pop-up menu: Remove Group, Edit Group and Group Information. 6.2 Remove Group Remove the group from the system by clicking the right mouse button and selecting Remove Group from the displayed menu. The group is removed and the channels that belong to that group turn gray. 6.3 Edit Group This option displays the Group Settings screen, with six tabs, where the special group functions as preparation of working standards, pre-dilutions, automatic post-dilutions, spike standard settings and CLP actions are set (see Figure 6.2). Figure 6.2 Group info Group Details The screen displays by default the Group Detail tab with the name of the group and the chemistries (and methods) belonging to that group, when the option is opened. Warning: In the case a diluter is present on the sampler and runs in overlapping mode are performed, all dilution options (working standards, pre- and post-dilutions) must be unchecked, as overlapping analysis and dilutions cannot be combined.

109 FlowAccess Chapter 6 Groups Working Standards settings To be able to prepare working standards, the methods either need to share the same standards or multi stock locations must be entered. When all standards are shared, the ratio for lowest to highest standard will be the same for all methods in the group. Check the standard range as set-up in the methods. A. When all methods share the same standards: 1. Click the Working standard settings tab (see Figure 6.3). Figure 6.3 Working standards 2. Check the Prepare working standards option. 3. Enter the position of the stock solution. (Do not place the stock on a position reserved for a working standard in the table). 4. Enter the stock concentration as a dilution factor for the highest standard. 5. If required, check Mixing. When Mixing is selected, some air is drawn up into the needle and then a part of the diluted solution. Then air and solution are pushed back into the tube, thus achieving better mixing. 6. Enter the number of times of mixing. 7. Click Save to accept the settings and to close the screen. Note: The total dilution volume is set in the tray selection box. Default for the sample tubes is 10 ml and for the standard tray 20 ml. Note: When a SA1070 sampler is used the working standards as well as the stock standard have to be placed in the sample racks, as the diluter needle cannot reach the standard containers.

110 FlowAccess V3 6-4 B. When the methods do not share the same standards: 1. Check the Prepare working standards option. 2. Check the Multi stock location option. The fields for the Stock location and Stock concentration are then not available anymore. 3. Click Settings. The Multi stock location screen is opened (see Figure 6.4). Figure 6.4 Multi stock locations 4. Enter the stock location and stock concentration (ratio) for a method and select the method from the list. See Figure 6.4 for an example. 5. Do this for each method. The methods that share the same stock location must have the same stock concentration ratio. Note: In the method setup the methods with the same stock location must share the same standard series. E.g. ammonia and phosphate in Figure 6.4 share S1, S2, S3 and S4 and nitrate+nitrite and nitrite share S5, S6, S7 and S8. 6. Press Save to close the screen. 7. If required, check Mixing. When Mixing is selected, some air is drawn up into the needle and then a part of the diluted solution. Then air and solution are pushed back into the tube, thus achieving better mixing. 8. Enter the number of times of mixing. 9. Click Save to accept the settings and to close the screen Pre-dilution settings 1. If automatic pre-dilutions are to be made on samples, click the Predilution settings tab. 2. If mixing of the pre-diluted samples is required check Mixing. 3. When this option is selected, enter the number of times the sample has to be mixed Spiking Standard settings 1. If samples are spiked, click the Spiking standard settings tab. 2. If mixing of the spiked samples is required check Mixing. 3. When this option is selected, enter the number of times the sample has to be mixed.

111 FlowAccess Chapter 6 Groups Post-Dilution settings 1. Click the Post-Dilution settings tab (see Figure 6.5) for advanced settings for automatic post-dilutions (also check Post-dilution in the methods, see chapter 5.3.6). Figure 6.5 Post dilution settings 2. Check the post-dilution option. 3. Click on Interval settings. The Post Dilution Batch Details screen is opened (see Figure 6.6). Figure 6.6 Batch details

112 FlowAccess V Set the batch size. Use the same batch size as in the original table. 5. Select the required sample types from the drop-down list in the left column (see Figure 6.6). At least a drift and wash are required for corrections. When selected the Identity is placed in the right column. Important: When Multiple peak detection is used, a Drift ignore (DI) is required as start of the post-dilution sequence. The sample types as set in the Batch Details must also be present in the basic table as information about the position of this sample types is required. The cup position taken for these sample types will be the last position for this sample type in the basic table. It is the operator s responsibility to make sure that these cup positions contain enough volume for the post dilution (e.g. take a new cup position for the last batch, see chapter ). Automatically the post dilution run will end with a drift and a wash. Important: The interval settings are related to the sample table and must be set for each new table 6. Click Save to save the settings and to close this screen. 7. Check or select the other options in the screen as required. The options are explained below. Re-samples The number selected here (max. = 9) is the number of sample cups following the over range sample that are re-analyzed during the post dilution run. Samples after an over range sample can be calculated incorrect due to the fact that the signal was not yet returned to the baseline when the peak of the sample started. Cascade The number of times a post-diluted sample has to be re-diluted when it is over range again (Max. = 9). Post dilution factor A selection can be made for the post-dilution factor to be used. - Use PD factor from methods the software looks for the postdilution factors in the methods that are over range and takes the highest dilution factor set in these methods. - Use this Post-dilution factor this option overrules the factors as set in the methods. When this option is selected a dilution factor has to be entered (selectable from 2 to 50 in whole digits). No resample of over range cups When this option is selected the over range sample after an over range sample is only post-diluted and not resampled. Use Mixing The sample is mixed when post-diluted. The number of times for mixing has to be entered. Note: Post-dilutions are always made from the last (available) position in the sampler. When post-dilutions are expected, put enough empty tubes in the sampler for the post-dilutions. In the case pre-dilutions were carried out in the sampler, the software looks for the last position before those pre-dilutions. Only when (large) pre-dilutions are carried out separately in advance of a run (see chapter ),

113 FlowAccess Chapter 6 Groups 6-7 these (intermediate) tubes have to be removed before starting the run as the sampler does not remember these positions. Operating principle for post-dilutions: 1. The run-out wash of the main run is marked for all chemistries that were started at the same time. 2. The program checks whether the first chemistry channel has the post dilution option selected in its method. If not, the program proceeds with step The unknown cups are checked for over range based on the criteria selected in the method (see chapter 5.3.4). 4. If the result of a cup is over range, the cup is added to a table with details like Location, Position, Type, Post dilution position, Post dilution factor etc. If no post dilution position exists in the table, that cup is not added to the table. 5. The re-sample cups are also added to the table. 6. Steps 3, 4 and 5 are repeated for all unknown cups for that chemistry. 7. If two subsequent cups are over range, the second cup is analyzed twice, once diluted and once undiluted (as re-sample, unless set different). 8. Steps 2 till 7 are repeated for all the channels. 9. Even if a cup is over range for more than one channel it will be diluted only once. 10. The dilutions will be made for all the cups except the re-sample cups. 11. After the dilutions are made the above table will be added to the main table with the drift/wash intervals as set in the Batch details. 12. The post-dilution run will start and end with a drift/wash. a. Example of a post dilution without re-samples. Assume that there are two channels running and both have over range cups (marked as x ). Cups Channel 1 Channel 2 Post dilution over range over range position A1 x D30 A2 x D29 A3 x D35 A4 x D28 A5 x x D34 B1 x D27 B2 x D33 B3 x D26 B4 x D32 B5 x D31 FlowAccessV3 first checks channel 1 for over range. For the over range cups A3, A5, B2, B4 and B5 the post dilution positions will be allocated from D35-D31. FlowAccessV3 then checks for channel 2 and allocates positions D30- D26. A5 will not be included because it is already included.

114 FlowAccess V3 6-8 So the dilutions will be made in the following order: A1 -> D30 A2 -> D29 A3 -> D35 A4 -> D28 A5 -> D34 B1 -> D27 B2 -> D33 B3 -> D26 B4 -> D32 B5 -> D31 The sampling (between the set batch interval details) will be made in the following order: D30, D29, D35, D28, D34, D27, D33, D26, D32, and D31. b. The following example describes a post dilution with re-samples. Consider the following example: Cup Over range Post dilution position A1 x D35 A2 x D34 A3 A4 x D33 A5 x D32 If the number of re-samples is two, the resultant table will look like the following. (Of course, with the set batch interval details) D35 Over range (A1) diluted A2 Resample A3 Resample D34 Over range (A2) diluted A3 Resample A4 Resample D33 Over range (A4) diluted A5 Resample D32 Over range (A5) diluted From the above example note that for D33 (A4) only one cup is added as re-sample. The reason is obvious. There are no cups after A5. Similarly for D32 (A5) no resample cups are added because there are no more cups after that.

115 FlowAccess Chapter 6 Groups CLP Action Fail status If CLP was checked in the methods, here the action for all methods in the group has to be set if the results are outside the set limits. Click the tab CLP Action Fail Status (see Figure 6.7). Figure 6.7 CLP Action Fail Status Check the option CLP Action Fail Status and click Settings to open the screen (see Figure 6.8) to set the actions to be taken when the CLP samples are outside the set limits. It is possible to enter up to 4 actions for items out of limits. Figure 6.8 CLP screen When clicking in the fields behind an item, the possible actions to be taken can be selected. When an CLP item is calculated to be outside the limits as set for a method, the analyzer checks for the required action and performs this action for the whole group. Dependent on the selected action for an CLP item, one or more fields can be filled with an action for that item.

116 FlowAccess V Example: for the ICV four actions are available Re-run ICV Re-run Standards Abort run and show messages Continue run and flag results For Action Fail status 1 and Action Fail Status 2 the actions Re-run ICV respectively Continue run and flag results are selected (see Figure 6.9). The following actions take place: 1. The run is stopped when the result of a method is outside the pre-set limits of the ICV for a method and the analyzer waits until the samples already taken from the table are detected in the Real Time screen. 2. The ICV is re-run and the run is resumed starting from the sample in the table after the ICV. 3. When the result is again outside the pre-set limits, the run is continued and the result is flagged. 6.4 Group Information When this option is selected the Group information dialog box is displayed with information about the group (see Figure 6.10). The box show the name of the group and the modules with the attached methods. Click Close to close the box. Figure 6.9 CLP actions Figure 6.10

117 FlowAccess V3 CHAPTER 7: PREPARING FOR ANALYSIS Preface: This chapter gives detailed instructions about the last checks and results format before for the analysis run is started.

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119 Chapter 7 Preparing for Analysis PREPARING FOR ANALYSIS When the setup of the method(s), group(s) and table(s) is performed as outlined in chapter 4, 5 and 6 the system seems ready for analysis. However, before an analysis is started, some analysis conditions have to be checked or set. - Check that all hardware is turned on (refer to the Operating the SFA Analyzer for the sequence). In FlowAccessV3 this can be done for each piece of hardware individually by means of the Remote control button or grouped by means of the Status button (see chapter 3.4). - Select a group for analysis, see chapter 7.1 (not when the time program is used). - Set up the digital matrix detector (if present and in combination with a 8503 or 8505 interface), see chapter Set up a time program (if this is to be used), see chapter Check the baseline in the Real Time screen, see chapter If overlapping analysis will be performed, make sure that this option is checked in Preferences, General Settings (see chapter ). For explanation of overlapping analyses, see chapter 8.7. Note: An extra feature is to start an analysis with Screening. For using this feature, a system controller must be present on the analyzer system as well as user defined macros and extra valves. This feature is explained in chapter Selection Before the groups can be selected for analysis or baseline checking, all hardware has to be turned on. Note: Remember that the system needs some time to stabilize. 1. Click on the Baseline or Analysis button on the toolbar. The check boxes behind the channels and before the name of a group become available and can be clicked to check them. 2. Select the group(s) to be analyzed by clicking the box before the name of the group. As soon as a group is selected the Start button on the toolbar turns yellow, which means that the button can be clicked to start an analysis or to view the baseline. The RealTime screen (see Chapter 8.7) can be opened to view the baseline. 3. It is possible to select each channel individually by clicking the check box of the channel. As soon as one channel of a group is checked, the check box of the group to which the channel is attached will be checked.

120 FlowAccess V Baseline checking Before the analysis is started the baseline can be checked in the RealTime screen. Figure 7.1 Baseline started Select the Baseline button and then check the group of which the baseline is to be viewed. Click the Start button on the toolbar; the status will be displayed in the right part of the screen (see Figure 7.1). Note: In the case that not all parts of the group are on, a message is displayed which items are not on and whether the analysis can be started when these items are not on. Select to start or not. Select the Realtime button to display the Real Time screen. Now the baseline of the selected chemistries can be viewed and adapted. All channels can be displayed in the same screen in a tiled view (see Figure 7.2). Figure 7.2 Baseline checking

121 Chapter 7 Preparing for Analysis 7-3 The screen shows in this case a four-channel system with the first channel selected. Double click on the channel to display it separately. Then all functions of the screen are available (see Figure 7.3). Figure 7.3 Baseline one channel The Y-scale can be adjusted for each channel. Adjustment of the X-scale changes the scale for all channels. To change the scale for the selected channel: 1. Click on the X or Y scale. The Properties dialog box is displayed with the scale of the selected channel. See Figure Change the Y-scale to the limits as desired. 3. If desired also change the limits for the X-scale. These changes will be applied for all channels in the system. 4. Click Save & Close. 5. To change the Y-scale for the second method, select the method from the Channels menu. The method is then displayed. 6. Again click on one of the-scales to display the Properties dialog box. 7. Change the Y-scale to the limits Figure 7.4 as desired. 8. In this dialog box also the Panning can be changed. By default the Panning is set on 25%, which means that the screen will jump 25%, when the peaks have reached the end of the screen during the run.

122 FlowAccess V If required the colors of Axis, grid and cursor can be changed. 10. Click Save & Close. 11. Do the same for each channel in the group. 12. When digital matrix detector(s) are present follow the procedure in Chapter When all limits are set according your wishes, there are two options: 1. Close the Real time screen by selecting Exit from the File menu. You are returned to the Active System screen where the analysis can be started later. 2. Start the Analysis directly from the baseline view (see chapter??). Restrictions is that it is not possible to enter a name for the analysis, other than the standard name generated by the system. 14. Select the Stop button and uncheck the group and click Yes to confirm the stopping of each channel. 7.3 Digital matrix detector setup When one or more digital matrix detector(s) are present in your system, this detector was setup in the system setup, see chapter and Offset Set the offset as required. Always set the offset of the measurement channel (blue line) at least 100,000 DU (0.1 AU) higher as the correction channel (red line). These offset settings are stored in the database. During data collection, the offset will be added to the data received and the corrected values will be stored in the real-time database. Factor The factor compensates the difference in peak height between the measurement channel and correction channel, due to the matrix effects. This factor must be established before the first sample run is started The factor and the offset for this matrix detector must be entered in the RealTime screen. Procedure: 1. Run the baseline in the RealTime screen (see Figure 7.5). Figure 7.5 A-B Corrected channel

123 Chapter 7 Preparing for Analysis Select the Display Matrix channels icon (most right icon) in the toolbar. This icon is only available when a matrix detector is present. The screen as displayed in Figure 7.6 is opened with the baseline of both channels. Make sure that the baseline of the measurement channel is higher than the baseline of the correction channel. Figure 7.6 Channel A and channel B 3. Select the Matrix Settings icon from the toolbar (see Figure 7.7). 4. Enter the offset for the correction channel to make the difference about 0.1 AU and click Save. 5. Close the screen. 6. Press the Stop button in Active system screen, uncheck the group and click Yes to stop each channel. Procedure for a salinity correction in seawater: 1. Prepare a table with at least three seawater samples and start an analysis (see chapter 8.1). Measure the samples without coloring agent, i.e. do not add hypochlorite for the ammonia reagent, or do not add ascorbic acid for the phosphate determination. All other reagents remain as to create similar ph and buffer conditions (use distilled water as rinsing liquid). Figure 7.7

124 FlowAccess V During the analysis measure in the Realtime screen the height of the displayed salt peak of channel A and B. Figure 7.8 Calculation of factor 3. Divide these 2 results A / B; the value obtained is the factor to be entered for the correction channel. 4. Enter the calculated factor in the Matrix Settings screen for the correction. The measurement channel is not corrected, since A is divided over B. 5. Click Save to accept the settings and to close the dialog box. 6. Check if the values of the next peak are equal. 7.4 Time programming Time programming is only possible when a system controller is present on the system. When the time program is used no chemistries or groups may be selected for analysis. The status setting is used for start up and shutdown of the system. It is not necessary to load a table in the sampler, as the table is selected in the time program. The programs are defined in the Time Program screen Time program Entry Select Time program entry from Time Program in the Instrument menu. The Time Program screen is displayed (see Figure 7.9). The fields are only accessible after selecting the New or Open button. Two types of time programming are possible: - Normal, for a complete start-up and/or shut-down program to be enabled before an analysis is started. - To be enabled after an analysis, for a shut-down and/or start-up and shut-down program to be enabled after the current run is finished.

125 Chapter 7 Preparing for Analysis 7-7 The screen shows seven columns for setting a complete program for automatic start-up and shutdown of an analysis or more analyses. Figure 7.9 Time program screen - The Day column is used to set the day of the week the analysis has to be performed. All days of the week are available. - The Time column is used to set the exact time for all actions to be performed for the analysis. The time can be set from 00:00:00 till 23:59:59. - The Hardware column is used to select either the system or a part of the system for which the procedure is set. - The Hardware number column is used to select a single item on the system (for example a certain rinsing valve). - The Procedure column is used to set the appropriate action. The available actions are: Change to Default status Change to Standby status Change to Online status Start Analysis Shut down Wait time (only visible after selection of Start Analysis ) - The Wait time column is used to set a waiting time between two analyses, as after the start of an analysis a relative time scale is used (a wait time can only be filled in when the Wait time option is selected). - The Analysis settings column (this column can only be filled on the row where Start Analysis or Shut down is selected) opens either a dialog box for the sampler and chemistry selection or pump pulse. Sampler and chemistry selection dialog box: 1. Select the chemistries of the system on which the analysis is performed. All chemistries on the active system are available for analysis. 2. Select the table to be used for the analysis. Only tables created for the active system are available.

126 FlowAccess V3 7-8 Pump pulse dialog box: 1. Check pulse pump. 2. Enter the time for the Pump On interval and the Pump On time. Note: Make sure you have finished your time program and enabled the time program before the time of the first instruction in your program has been passed. Otherwise the program skips this instruction and takes the next one. In the case of two samplers in one system a time program can only be used for either both samplers at the same time, or for one sampler, when they are not to be started at the same time. However, when two systems are present, it is possible to use a (different) time program for both systems independently. It is also possible to set a secondary time program to be enabled after the first set program e.g. for Wake-up in the morning after a shutdown time program of the current run. The instructions for both time programs cannot be put in one time program as the time in a program is either definite or relative. Four examples are explained with time programming: Program A To perform an analysis that takes about three hours, to be started early in the morning so that the analysis is finished when you arrive at work. Program B To perform a shut-down of an analysis that has been started during office hours and running until after office hours. Program C To start automatically an analysis in the high (low) range after a current run in the low (high) range. Program D To perform a Wake up program that is set as secondary program.. To create time program A: Task: The analysis has to be started at about five o clock in the morning in order to be finished at about eight o clock. In this case the system will be shut down first in the evening to save reagents and wake up in the morning for analysis. Make sure the system is in Default Status (rinsing valves on water) or use that as instruction in your time program. Note: When using the time programming it is important to use the different status settings of the system. Make sure that the status settings are correct to start up the system (see also chapter 2.2.3). 1. Select New from the File menu or from the toolbar. 2. Select Normal and type a name for the program (see Figure 7.10). 3. Click OK to close the screen. Now the fields are available in the Time program screen.

127 Chapter 7 Preparing for Analysis 7-9 Figure Click the down arrow in the Day column and select the current day (Tuesday). 5. Select the cell in the Time column and type the time the system has shut down (17:00). It is also possible to double click on the cell. An up and a down arrow will be displayed. The time can then be selected with the arrows. 6. Select the cell in the column Hardware and click on the down arrow to display the list with hardware. Select System. 7. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Shut down. A gray button called Settings becomes available. 8. Click the Settings button to open the Pulse pump dialog box. Check the pulse pump option and type the time interval for working of the pump and the time the pump has to be working (see Figure 7.11). Figure Go to the next row and select the next day (Wednesday). For the time select for example 04:40. Select for the hardware again System. 10. Select for the procedure Change to Standby Status. 11. Go to the next row and repeat step 9. Select for the time 04: Select for the procedure Change to Online Status. 13. Go to the next row and repeat step 9. Select for the time 05:00

128 FlowAccess V Select for the procedure Start Analysis. At this moment the gray Settings button will be displayed in the Analysis Settings column. Click on this button to open the Sampler and Chemistry Selection dialog box. Here the options for the analysis are selected (see Figure 7.12). Figure 7.12 Select Chemistries and sampler 15. Check the chemistries for which the analysis has to be performed. (Remember not to check the chemistries in the Active System screen). 16. As soon as one chemistry is checked a table can be selected for the analysis. Click on the down arrow and select the table for the analysis from the list of available tables for the system. 17. Select Save from the File menu to save the settings and to close the box. Your Time Program screen will now look like Figure Figure 7.13 Creating Time Program A

129 Chapter 7 Preparing for Analysis 7-11 To create time program B. Task: The analysis was started in the afternoon and has to be shut down when the analysis is ready. Note: When using the time programming it is important to use the different status settings of the system to make the system ready to perform an analysis while saving as much energy and reagent as possible. Make sure that the status settings are correct to start up the system (see also chapter 2.2.1). 1. Select New from the File menu or from the toolbar 2. Select the option To be enabled after an Analysis and type a name for the program. 3. Click the down arrow in the Day column and select the current day. For example Tuesday. 4. Select the cell in the Time column. The cell will automatically display Relative. 5. Select the cell in the column Hardware and click on the down arrow to display the list with hardware. Select System. 6. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Wait time and enter the time the system has to wait before changing to the next status. For example 2 minutes. 7. Go to the next row and repeat step 3, 4 and Select for the procedure column Change to Standby status. 9. Go to the next row and repeat step 3, 4 and 5. Select Wait time for the procedure and enter a waiting time. For example 2 minutes. 10. Go to the next row and repeat step 3, 4 and 5. Select for the procedure Change to Default status. 11. Go to the next row and repeat step 3, 4 and 5. Select for the procedure Wait time and type a waiting time for the system. If for the Default status is defined that the valves are set on water, the system is rinsed during this time. For this example we use 15 minutes. 12. Go to the next row and repeat step 3, 4 and Select for the procedure Shut down. A gray button called Settings becomes available. Figure 7.14 Creating Time Program B

130 FlowAccess V Click the Settings button to open the Pulse pump dialog box. Check the pulse pump option and type the time interval for working of the pump and the time the pump has to be working (see Figure 7.11). The screen should now look like Figure Click Save. Close the screen. Enable the program during the analysis run (see chapter 7.4.2). 16. After finishing the run the program will go into the Shutdown mode and the pump will run at the time interval as set in the program. The program stays open and the Stop button will be available. Press this button to make the system available for new actions (see Figure 7.15). Figure 7.15 Note: Do not stop the program by pressing the red cross, as the pumps will not be in the correct status when the program is re-opened. To create time program C. Task: To start an analysis in the high (low) range when the current run in the low (high) range is finished. Note: The pre-dilution loop must be connected to the system controller as a normal rinsing valve (R1-6). 1. Start the run in the high (low) range with the pre-dilution loop set the correct position (reagent or water). 2. Select New from the File menu or from the toolbar. 3. Select the option To be enabled after an Analysis and type a name for the program. 4. Click the down arrow in the Day column and select the current day. For example Monday. 5. Select the cell in the Time column. The cell will automatically display Relative. 6. Select the cell in the column Hardware and click on the down arrow to display the list with hardware. Select System. 7. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Wait time and enter the time the system has to wait before changing to the next status. For example 2 minutes. 8. Go to the next row and repeat step 4, 5 and 6. By default the same day and relative time as on the previous row is selected. 9. Select the cell in the column Hardware and click on the down arrow to display the list with items that can be selected. Select Rinsing valve.

131 Chapter 7 Preparing for Analysis Select the cell in the column Hardware number and click on the down arrow to display the list with the valves that can be selected. Select the valve number that is used as pre-dilution loop. 11. Select the cell in the column Procedure and click on the down arrow to display the selectable items (reagent and water). Select the opposite position as used for the current analysis. 12. Go to the next row and repeat step 4, 5 and 6. By default the same day and relative time as on the previous row is selected. 13. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Wait time and enter the time the system has to wait before changing to the next status. For example 2 minutes. 14. Go to the next row and repeat step 4, 5 and 6. By default the same day and relative time as on the previous row is selected. 15. Select the cell in the column Procedure and click on the down arrow and select Start Analysis from the list. 16. Select the cell in the column Analysis Settings and click on the down arrow and select Start Analysis from the list. At this moment the gray Settings button will be displayed in the Analysis Settings column. Click on this button to open the Sampler and Chemistry Selection dialog box. Here the options for the analysis are selected (see Figure 7.16). Figure 7.16 Select chemistry 17. Check the chemistry for which the analysis has to be performed. (Remember not to check this chemistry in the Active System screen). 18. As soon as the chemistry is checked a table can be selected for the analysis. Click on the down arrow and select the table for the analysis from the list of available tables for the system. 19. If desired, type a name for the analysis and click OK to close the dialog box. Your Time Program screen will now look like Figure 7.17.

132 FlowAccess V Figure 7.17 Creating Time Program C To set time program D: Task: To perform a Wake Up of the system as a secondary program to be performed after an analysis shutdown. 1. Select New from the File menu or from the toolbar. 2. Select the option To be enabled after an Analysis and type a name for the program (e.g. Wakeup, see Figure 7.18). Figure Click the down arrow in the Day column and select SMTWTFS. When all days are selected the program can be used for all days as later, when the program is used, a specific date and time is entered. 4. Select the cell in the Time column. The cell will automatically display Relative. 5. Select the cell in the column Hardware and click on the down arrow to display the list with hardware. Select System. 6. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Wait time and enter the time the system has to wait before changing to the next status. For example 2 minutes. 7. Go to the next row and repeat step 3, 4 and 5 8. Select the cell in the column Procedure and click on the down arrow to display the list with procedures that can be performed. Select Change to Standby status. 9. Go to the next row and repeat step 3, 4 and 5.

133 Chapter 7 Preparing for Analysis Select for the procedure column Wait time and enter the time the system has to wait before changing to the next status. For example 15 minutes. 11. Go to the next row and repeat step 3, 4 and Select for the procedure column Change to Online status. Note: The procedure is an example and is dependent on how the different status were defined. Note: Make sure that the first time program is finished before the secondary time program is to be started. Otherwise the program skips this program and starts the next day Enable a Time Program 1. Select Time Program Selection from Time Program in the Instrument menu. 2. Select the type of program and the name of the program (see Figure 7.19) 3. If the time Program is to be enabled immediately, check the option. 4. Click OK to close the screen. The Time Program is enabled. Figure 7.19 Time program selection Set secondary time program As soon as a time program as program B is enabled, it is possible to set a secondary program as program D. 1. Select the option Set secondary time program from Time Program in the Instrument menu. The dialog box is displayed (see Figure 7.20). Figure 7.20

134 FlowAccess V The running time program is displayed (Endsystem1). Select the second program (Wakeup1). 3. Set the start date and time for the start of the second time program. The relative time in the time program will be changed into the set day and time. Make sure that the first time program will have been ended, before the second program is started. 4. Select Set to set the second time program. An example of a set secondary time program (with 2 systems) is shown in the dashboard of Figure Figure Preparing a run from an external macro Note: This feature requires user defined macros and is only available on request! It is possible to start a run from an external macro. Macros can be used for several purposes. For example for Overlapping analysis and for Screening. The system hardware must be suitable to be able to run an external macro and there are special restrictions when using a macro. To use this feature the option Allow access of an External Application must be checked in Preferences, General Settings, see chapter a. Overlapping run To use this feature also the option Allow to run in Overlapping mode must be checked in Preferences, General Settings, see chapter The option Overlapping analysis using an external macro is described in chapter 8.7. b. Screening To use this feature also the option Execute the external application must be checked and set in Preferences, General Settings, see chapter

135 Chapter 7 Preparing for Analysis 7-17 Screening is meant to sort out the global concentration of a parameter (e.g. Total Cyanide) in a quick run on high pump speed, where after the run is repeated under normal conditions with the samples sorted from low to high in a new table to avoid contamination and/or long rinsing times. A third run can be done on free Cyanide for the samples with high concentration. When a macro is used for Screening, the system must be provided with a system controller and extra valves to be able to switch automatically between types of analysis. For an example of a flowchart hoe to run this feature see Appendix, H.

136

137 FlowAccess V3 CHAPTER 8: ANALYSIS RUN Preface: This chapter gives detailed instructions about all aspects of analysis run

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139 Chapter 8 The Analysis run ANALYSIS RUN 8.1 Starting the analysis run When methods, tables and groups are defined for the Active System and, if required, the last preparations as described in chapter 7, an analysis can be started. A. Start from the Active System screen. After viewing (and adapting, if required) the baseline, the view was closed and returned to the Active System screen. The run gets by default a name with the format as defined in the File Settings tab of the Preferences menu in the Main screen, but another name can be given to the analysis (see Figure 8.1). The file will be saved in the directory as set in Path Settings tab. To start an analysis: 1. Select the Analysis button and then check the group for which the analysis has to be started. Figure 8.1 Save run file name 2. Select the Start button on the toolbar; accept the run file name as displayed in the pop-up screen (see Figure 8.1) or type a new name for the run file and click OK.

140 FlowAccess V3 8-2 Note: In the case that not all parts of the group are on, a message is displayed which items are not on and whether the analysis can be started when these items are not on. Select to start or not. 3. The analysis is started and the status ( Started ) will be displayed in the right part of the screen (in the case that 21CFR part 11 is active first the operator name has to be entered). 4. If automatic pre-dilutions are present in the table, the sampler will start with the making of the pre-dilutions. Then the actual run will start. 5. Select the Results button to view the incoming results. The sample table can also be viewed and even be edited during the run. Select the Realtime button to view the peaks and the progress of the run. When Realtime is selected immediately after starting the run, a screen is displayed that data is being collected (see Figure 8.2) until data can be displayed. Figure 8.2 Note: The display of the Results screen can be adapted by changing the settings in the Format Fields tab (see chapter 8.5). B. Start directly from the Realtime screen in the Baseline View The analysis can be started directly from the Realtime screen where the baseline was viewed (and adapted, if required). 1. Select Quick Start from the File menu (see Figure 8.3). The Quick Start dialog box is displayed (see Figure 8.4) Figure 8.3 Quick Start option

141 Chapter 8 The Analysis run 8-3 Figure 8.4 Channel selection 2. The sampler and the channels that are displayed in the baseline view are displayed and selected. Deselect the channels that are not to be started for the analysis and select the Start button. 3. The run is started with the default name, selected by the system, and the dialog box is closed as well as the Realtime screen. You are returned to the Active System screen, where the dashboard shows that the system is in Analysis 8.2 Viewing incoming results The incoming results can be viewed in the Results screen. The Results screen can be opened as soon as the analysis has been started. The screen is displayed with four tabs: Detailed Results, Calibration, Statistics and Edit Table & Methods. The tab Detailed Results will be displayed (see Figure 8.5). Figure 8.5 Incoming results

142 FlowAccess V3 8-4 The results are displayed for all channels in one screen. If one or more Flag column is selected in Format Fields (see chapter 8.5), the flags are displayed as in the Peak Information screen. Three types of flags are possible: Peak flags: N Normal peak (peak within the measuring window) E Early peak (peak before the measuring window) L Late peak (peak after the measuring window) A Absent peak (no peak found in the measuring window higher than height reject) D Diluted Peak (only displayed when post-dilution is performed) O Over range Peak (peak outside the measuring limits, automatic post-dilution performed, if post-dilution is checked) [W] Warning (only in case of multiple peak picking) that one of the peak detection methods was outside the tolerance limit and excluded from peak calculation. [E] Error (only in case of multiple peak picking) that more than one of the peak detection methods was outside the tolerance limit and excluded from peak calculation. CLP flags: Reports the Action Fail Status (if applicable) of the CLP sample. Results flags: A(L) Lower Alarm (peak lower than the limit as set in the methods for the QC s and unknown samples) A(U) Upper Alarm (peak higher than the limit as set in the methods for the QC s and unknown samples) X Excluded from calculation (only for Standards, Drifts and Washes) A combination of flags is possible, for example D, N. This means that this is a peak of a post-dilution run, which is measured as normal.

143 Chapter 8 The Analysis run Viewing the calibration curves As soon as the drift and wash after the standards have been measured, the calibration curve is calculated and displayed in the Calibration tab (see Figure 8.6). Figure 8.6 Calibration curves The calibration curves are displayed with a max. of 4 channels in one screen. If more channels are being measured, the remaining channels can be viewed by pressing the Next button. The type of calibration (as set in the method) is displayed for each channel. The calibration curve for all other calibration types can be viewed by selecting the calibration type from the drop-down menu.

144 FlowAccess V3 8-6 A calibration view can be extended by pulling out the X and Y borders of the calibration curve (see Figure 8.7). Figure 8.7 If Auto calibration is selected the best fitting calibration curve is displayed with the name of the selected calibration type. The calibration constants are displayed below each calibration curve. a (Intercept) is the intercept of the Y-axis. b (Slope) is the regression coefficient, which stands for the sensitivity of the method. c (Constant) is a coefficient of the 2 nd order calibration. r (Correlation) is the relation between the calculated concentration and the theoretical concentration of the standards. R 2 (R-squared) Statistical measure of how well a regression line approximates real data points; an r-squared of 1.0 (100%) indicates a perfect fit. RSD (Residual standard deviation) describes the scatter of the values about the calculated regression line, describing the precision of the calibration. Editing of calibration curves is explained in chapter Viewing the run in Realtime About 10 seconds after the analysis is started the course of the run can be viewed in Realtime. By default all channels are displayed in one screen in Tile view (see Figure 8.8). This view is convenient to overview the course of the run. In this view no adaptations can be made on the display properties. This is done for each channel separately.

145 Chapter 8 The Analysis run 8-7 Figure 8.8 Realtime view of 4 channels To view a channel separately: Click in the channel to make it active and click on the Maximize tool or double click on the title bar. The channel is now displayed in full screen where all display and information tools are available (see Figure 8.9). Figure 8.9 Realtime view of one channel The display of the screen can be changed: zoom in and zoom out, the X and Y values and display properties. The grid, peak labels and measurement window can be displayed. Also information about the peaks (see Figure 8.10) can be retrieved. View settings are saved when closing the screen and retrieved at re-opening.

146 FlowAccess V3 8-8 Figure 8.10 Peak information By means of the arrows one can toggle easily between the peaks and view the information. The option Exclude from Calculation is meant only for the Standard, Drift, Wash and QC samples to exclude these results from correction and validation (see also chapter 8.6.1). The view of the Peak information dialog box is dependent on the selected peak detection settings in the Method Settings. Note: During analysis it is strongly advised not to change the peak detection settings. During analysis, one can select Other Settings from the menu bar in Active System and then select Analysis information. After 3 measured peaks the expected end time of the run (without post-dilutions) is calculated and displayed (see Figure 8.11). Figure 8.11 Expected end time of analysis

147 Chapter 8 The Analysis run Format Fields Result Format The view of the results can be customized by means of the Result Format screen (see Figure 8.12). This screen is opened by selecting the Format tool in the Results screen (or by selecting Format Fields from the Options menu). Figure 8.12 Result Format screen The screen is divided in fields. - The left field shows the columns that are displayed in the sample table. - The upper middle field shows the chemistries being analyzed. - The upper right field shows the available columns for each individual chemistry to be displayed in the Results screen. - The lower screens display custom formulas and custom chemistries. When the screen is opened the items are not available. To customize the view of the Results screen: 1. Click Add to make the fields available. 2. If more than one sampler is present, select the sampler for the view. 3. If more than one needle is present, select whether all needles or 1 should be displayed in the view. 4. Check in the left field the items to be displayed in the Results screen. By default position, type and identity are checked as they are always present in the sample table. 5. Select a chemistry in the middle field and then check the required options in the right field. 6. Repeat step 5 for all chemistries. 7. Click Save. A dialog box is displayed to type the name for the view. This name must have at least 4 characters starting with a letter. 8. Click Save.

148 FlowAccess V Select in Available Views by means of the drop-down menu the just created view from the list and select Show View. The screen is closed and the selected view is displayed in the Results screen. The name of the view is shown in a tab at the bottom of the screen. The original view can be displayed again by selecting the default tab. For example: a system with 2 needles is displayed by default with all needles in one view (the results of the needles are placed after each other, see Figure 8.13). Figure 8.13 View with 2 needles after each other The result of each needle can be viewed separately when these views were defined in Format results (see Figure 8.14 for second needle). Figure 8.14 View with second needle only

149 Chapter 8 The Analysis run 8-11 To edit a view: 1. Select the view to be edited from the drop-down list in the Available views and click Edit. The fields become available. 2. Change the selections as required in all fields. 3. Click Save. The Default view is shown in the Available views box. 4. Select the required view from the drop-down list and select Show View. The screen is closed and the edited view is displayed in the Results screen. To delete a view: 1. Select the button Delete. 2. A dialog box is displayed where the views to be deleted can be checked (see Figure 8.15). 3. Select the views to be deleted and click Delete. 4. In a pop-up screen the deletion has to be confirmed. 5. Click Cancel to close the dialog box. To set a default view: 1. Select Set Default Figure 8.15 Views at the bottom of the screen. The Select Views dialog box is displayed (see Figure 8.16). Figure 8.16 Select Views

150 FlowAccess V Select a view in the left column. 3. Click the upper arrow to move the view to the right column. 4. Click Save. 5. Click OK to close the dialog box. This view will now be displayed by default together with the default view (which is automatically displayed the first time the Results screen is opened), the next time the results are opened from the PostAnalysis screen. Only one view can be selected as default view. To remove a Default view: 1. Select Set Default Views at the bottom of the screen. The Select Views dialog box is displayed (see Figure 8.16). 2. Select a view in the right column. 3. Click the lower arrow to move the view to the left column. 4. Click Save. 5. Click OK to close the dialog box. This view will not be displayed anymore by default together with the default view the next time the results are opened from the PostAnalysis screen Custom chemistry If required extra columns can be added for a calculation result. This is called a Custom Chemistry. To add a Custom Chemistry: 1. Select the view, where a custom chemistry is to be added, from the list in the Available views field. 2. Select the Custom Chemistry tab at the top of the screen. The screen as shown in Figure 8.17 is displayed. In this screen the formula for the calculation of the customized chemistry can be entered. Figure 8.17 Direct formula

151 Chapter 8 The Analysis run Direct formula 3. In the left upper field the columns of the selected view are displayed. As soon as a custom formula is made active, the name of this formula will also be displayed in this field. 4. Two formula options are available: Direct formula - for simple calculations (like subtracting NO 2 channel from the NO 3 + NO 2 channel or blank channel correction) Beer calculation - for all aspects of a beer and wort analysis For example: An analysis run comprises the chemistries NO 3 + NO 2 and NO 2. To obtain the results for NO 3 the results of the columns with the results of the two chemistries can be subtracted with a correction for the molecule weight of NO 2. The results are directly calculated and displayed behind the last column of the original view in the Results table. Another option is to use the customized channel for the calculation of an analysis corrected for its blank analysis. The columns with the measured height of the analysis and the blank analysis are subtracted and the method is assigned to the customized chemistry where the calibration is defined. In this way the calibration is performed on the corrected heights. Two columns are added to the Results table. The first column displays the corrected heights according to the formula. The second column displays the calculated concentrations for the corrected heights. The decimal places to be displayed in the columns can be defined. To enter a formula: 1. Double click on an item in the left field (or click Add ), then double click (or click Add ) the required calculation item in the Operators field and again on an item in the left column to prepare a calculation formula. If numbers are required in the formula type the number in the numbers field and select Enter. For example: resultno3+no2-[1.4*resultno2] The entered formula will be displayed in the Formula field. If a formula for correction of a blank channel is created, then assign the customized channel to the measuring channel and select Set. Note: When a formula contains a division then the denominator may not be zero. In this case directly behind the division mark the function NULLIF must be double clicked in the right column and then the term inserted before the displayed 0. Example: custom.[module 7- Results] / NULLIF( custom.[module 6- Results],0) 2. Check the displayed formula if it is correct and click Save Formula and type a name for the formula in the dialog box. This will be the title of the new column in the results screen. If the formula is not correct click Clear Formula and start again. A formula cannot be edited. 3. Click OK. The name of the formula is now displayed in the left field. 4. Click the Result Format tab. The name of the formula will be displayed either in the custom formula list or in the custom chemistry list (when assigned to a method). 5. Click Show View. The calculated results are now displayed in the added column(s). To delete a formula: 1. Click Formula List. The Available Formula List screen is displayed (see Figure 8.18). 2. Select the formula to be deleted and select Delete or select Delete All.

152 FlowAccess V Confirm the deletion by selecting Yes in the dialog box. 4. Click Cancel to close the screen. The formula(s) will now be deleted from the left field and also from the Custom formula or Custom Chemistry field in the Result Format tab Beer Calculation Figure 8.18 Available formulas Several kinds of information are interesting in beer and wort analysis. In this dialog box eleven calculation formulas for information about beer and wort are available (see Figure 8.19): Figure 8.19 Beer calculation

153 Chapter 8 The Analysis run 8-15 Alcohol w/w; calculation of alcohol by weight Apparent Extract Content; calculation of the Apparent Extract, AE. Real Extract; calculation of the Real Extract, RE. Specific Gravity of Distillate, SG A. Balling; calculation of the Original Wort Extract, OwE ºPlato. Iteration; calculation of the original Apparent Extract, OAE. % Plato; calculation of percentage Original Wort Extract Real Degree of Fermentation, RDF% (formula according ASBC or EBC). Apparent Degree of Fermentation, ADF% (formula according ASBC or EBC) Abbreviations used: %Alc v/v = Alcohol percentage by Volume %Alc w = Alcohol percentage by Weight SG 20/20 = Specific Gravity, Density 20º/20º SG A = Specific Gravity of the distillate SG ER = Specific Gravity of the residue RE = Real Extract in ºPlato OwE = Original wort Extract in ºPlato AE = Apparent Extract OAE = Original Apparent Extract RDF% = Real Degree of Fermentation in percentage ADF% = Apparent Degree of Fermentation in percentage To obtain these results the following calculation formulas are used: Calculation of the percentage Alcohol by Weight: %Alc w = A p = %Alc v/v * 0.791/SG 20/20, whereby %Alc v/v = Result analyzed by SFA system, colorimetric method SG 20/20 = Density 20º/20º, de-carbonated beer, result SFA system with integrated PAAR DMA density meter. The direct reading of the density is corrected automatically from density 20º/4º to density 20º/20ºby dividing by This is called the apparent density. Calculation of the SG A from %Alc w 1. From table 2. With algorithm: A:=-1; B:= 1; SG A := 0; Count:= 1; Repeat Begin Res:=517.4 * SG A * Sqr(SG A ) * (SG A ) 3 -A p ; If Res > 0 then B:= SG A ; Else A:= SG A ; SG A := (A + B)/2; Count:= Count + 1; End; Until (Abs (A B) < ) Or Count > 100); SG A := 1 SG A ;

154 FlowAccess V Calculation of SG ER from SG A SG ER = 1 + SG 20/20 SG A Calculation of the Real Extract, RE RE = * SG ER * (SG ER ) 2 Calculation of the Original Wort Extract (Balling), OwE ºPlato Available in FlowAccess as specific beer calculation formula, called Balling OwE ºPlato = 100 * ( * %Alc w + RE) / ( * %Alc w ) Calculation of the Apparent Extract, AE AE = * SG 20/ * (SG 20/20 ) 2 Calculation of the Original Apparent Extract (by Iteration), OAE Available in FlowAccess as specific beer calculation formula, called Iteration OAE = AE + (AE RE)/ ( (0.001 * (AE + (AE RE)/0.23))) Calculation of the Real Degree of Fermentation, RDF% RDF% = 100 * ((OwE RE)/ OwE) * (1/1 ( * RE)) [=formula ASBC] Or RDF% = 100 * (( * %Alc w /( * %Alc w + RE)) [=formula EBC] Calculation of the Apparent Degree of Fermentation, ADF% ADF% = 100 * ((OwE AE)/ OwE) Or [=formula ASBC] ADF% = 100 * (( * %Alc w /( * %Alc w + AE)) [=formula EBC] Definitions: Standard Reference Wort (SRW): The reference for all subsequent quality and efficiency calculations as well as for component ratios. Extract: In the brewing sense encompasses the total available soluble mass in a brewing material as is, and/or potentially through processing. Original Extract: The total volume of wort as delivered to the kettle, containing the full complement of solutes, hopped, finished, boiling, as is, for transfer to the kettle. The Original Extract of the Wort (OwE) and of the Beer (ObE) may or may not be identical. Plato Value, total Extract: Original Extract of Standard Reference Wort. Apparent Extract: Measured value in de-carbonated beer expressing the amount of saccharose, which is dissolved in 100 g water at 20 ºC totally, which gives the identical density relative to the de-carbonated beer. Real Extract: Measured value in de-carbonated beer expressing the amount of saccharose, which is dissolved in 100 g water at 20 ºC totally, which gives the identical density relative to the de-carbonated beer and the de-alcoholized beer.

155 Chapter 8 The Analysis run 8-17 To display the calculated results for any item in the dialog box: 1. Enter the name for the column as it must be displayed in the results. 2. Select from the drop down lists the results columns of density and alcohol. 3. Enter the required number of decimal places to be displayed. 4. Select the required calculation item and click Save. The name of the column is displayed in the lower right field in the Results format tab. The calculation for the selected item is performed in the background and the results are displayed in the column in the Results screen of the selected view. 5. Follow step 1 to 4 for each calculation item in the Beer Calculation screen.

156 FlowAccess V Editing during the run Editing in Results screen In the Results screen (tab Detailed results ) the results are displayed as the corrected mean of the detection methods as set in the methods (see also chapter 5.3.2). Depending on the selected view (see chapter 8.6) also the flags are displayed (for example the results and peak flags for module 7 in Figure 8.20). Figure 8.20 Peak information When double clicking the result of a channel, the Peak Information dialog box is displayed (see Figure 8.20) with information about the peak. The Peak information box displays all relevant information about the sample, of which the peak is displayed. In this way it is easy to see if a peak was detected correctly according the pre-set peak detection method(s) by means of the flag info. When one of the detection methods was not within the set tolerance the peak flag shows a [W] for warning behind the peak flag (described in chapter 8.2) and that detection method was excluded from the calculation of the result. When more than one detection method was not within the set tolerance the peak flag shows an [E] for error behind the peak flag (described in chapter 8.2) and these detection methods were excluded from the calculation of the result. In the case of the above picture the peak shows the flag N,[W]. This means (see also chapter 8.2) that the peak is a normal peak and that one of the pre-set peak detection settings was outside the set tolerance limit and discarded from calculation. The option Automatic Peak detection is selected by default. Note: During analysis it is strongly advised not to change the peak detection settings.

157 Chapter 8 The Analysis run 8-19 The option Exclude from Calculation is meant only for the Standard, Drift, Wash and QC samples to exclude these results from correction and validation. A Results flag (Ex) will be placed in the results. When the option Exclude from Calculation is selected, the peak( result) is directly removed from the calculation of the calibration curve. By means of the button Calibration the list of used standards can be viewed (see Figure 8.22). When the result of a standard is not acceptable to be included, the result can easily be deselected from the calibration curve by deselecting the standard in this screen (same result as direct selection of the option Exclude from Calculation ). Press Save to close the screen. Note: Only when a standard was selected to view the info, the button Calibration is displayed Editing the calibration curve In the Results screen (tab Calibration ) the calibration curves of all channels are displayed, grouped by a max. of four calibration curves in a screen. (see Figure 8.21). Figure 8.21 By double clicking in a calibration curve, the list of standards is displayed (see Figure 8.22). When a standard is not acceptable to be included, it can be deselected. When pressing Save the screen is closed and the calibration curve is recalculated immediately with the new settings. Note: This change in the calibration curve can also be achieved by means of the Peak information screen opened from the results (see chapter 8.6.1)

158 FlowAccess V Figure 8.22 Select/deselect standards Table editing During the run it is possible to view and edit the sample table that is used for the analysis. The table can be opened from the Active System screen as well as from the Results screen. Note: It is not possible to edit the sample table during the run when the Time program is used. To open the table from the Active System screen: Select the Table icon and then Edit Table and click OK. To open the table from the Results screen: Select the tab Edit Table & Method. Select the sampler from the Table Entry list and click Edit. Editing of samples is only possible for the positions that are not yet sent to the sampler. As soon as the run is started, the first four cups are sent and cannot be edited anymore. The cups that are not available for editing anymore get a pink color (see Figure 8.23). Note: Do not use the Table Wizard to edit the table. Preferably add samples at the end of the table and add a drift/wash if more samples are added than the drift/wash interval in your original table. Samples can be inserted between existing samples by inserting rows. Make sure that changes in a sample are made and saved before the table has reached the changed or inserted sample. Note: If more than 1 needle is used in the table, and the table is edited, make sure to enter information for the other needle(s) as well. The table can be edited in the same way as before starting the analysis. Save the changes and return to the Active System screen, from where the Real time screen and the Results screen can be opened. Note: When a table was edited during the run and the table is loaded again for another run a dialog box is displayed to ask if the original settings should be kept.

159 Chapter 8 The Analysis run Method editing Figure 8.23 Sample table during run It is possible to edit some aspects of the method during the run, but not for damping. For example: one of the calibration standards appears to be not properly prepared and unusable for the calculation of the calibration curve. This calibration standard can then be deselected and will be excluded from the calibration curve. However it is easier to exclude a standard form the calibration curve immediately from the calibration curve or via the Peak information screen directly from the results. It is strongly advised not to change the peak detection settings during analysis Save the changes and close the Method screen. The changes are immediately visible. Note: When methods are changed during analysis and a next run is started, a dialog box pops up with the question if the original settings should be kept (see Figure 8.24). Figure 8.24

160 FlowAccess V Overlapping analyses It is possible to run overlapping analyses to save time when large sample loads are to be analyzed. Warning: There are a few restrictions when applying overlapping analyses 1. The diluter option cannot be used (no pre- and post-dilutions, no working standards) 2. The time program cannot be used 3. The same group and methods must be used. To activate this option check the option Allow to run in Overlapping mode in Preferences, General Options (see chapter ) before the first analysis is started. The principle of overlapping analyses is that as soon as the last sample of the run is taken by the sampler, a message is displayed that the trays can be changed. After a few minutes (built-in safety time) a message is displayed that the new run can be started. When the last sample of the previous run is calculated, the run is packed and saved and can be viewed in PostAnalysis. Overlapping analyses save thus almost twice the retention time of the chemistry modules. Two types of overlapping analysis are possible: 1. Using tables within FlowAccessV3 TM. 2. Optionally on request of the customer it is possible to combine Overlapping analyses' with importing tables from an external source. To run overlapping analyses with tables within FlowAccessV3 TM. 1. Make sure that the option in checked in Preferences. 2. Check the Start Ignore time in the Method settings. When the system has a long retention time (10-15 min), leave this time on 120 sec. When the system has a short retention time this time can be reduced to 50 seconds. 3. Prepare a sample table as described in chapter4.5 or Select a group for analysis and check the baseline as described in chapter 7.1 and chapter Start the first run as described in chapter While running the analysis prepare a new sample table for the next run from the Table menu. This table is not yet attached to the sampler. 7. Wait until the message Sampling completed, user can change tray is displayed (see Figure 8.25) in the task bar, exchange the sample trays from the sampler for the trays for which the new table was created or place the new samples in the trays according the new table.

161 Chapter 8 The Analysis run 8-23 Figure Wait until the message Allowed to start the overlapping run is displayed in the task bar. During this stage most menu items are not available (see Figure 8.26). Figure Remove the table attached to the sampler and load the just created table. 10. Start the next run as described in chapter 8.1.

162 FlowAccess V Note: When the Realtime screen was still open from the first run, the start of the new run is already displayed while the previous run was not yet closed. Then, when the first run is finished, the Realtime screen removes the displayed peaks of this run and continued with the peaks of the new run. The same counts for the results in the Results screen. The run is packed and saved and can be opened from the DataProcessing screen (see Figure 8.27). Figure At the end of the last run of the day, ignore the messages and wait until the run is completely finished, packed and saved. Remark: When it is not required that standards are analyzed in every run, it is possible to prepare the second table without standards. However the run will not be calculated during the analysis. When the run is finished, the run can be opened in PostAnalysis and the method changed into Calibration from a previous run with the first run selected. Then the results are calculated based on the calibration of the first run. To run overlapping analyses with tables from a macro: 1. Make sure that the options Allow to run in Overlapping mode and Allow access of an external application are checked in Preferences. 2. Select a group for analysis and check the baseline as described in chapter 7.1 and chapter Prepare a sample table in the external macro. Warning: When the system is new, before the first run is started, a table has to be prepared and saved as in the first table the trays on the sampler are defined. This information is required in Flowaccess for correct sampling. 4. Depending on the type of macro the run can be started directly from the macro. This macro must then contain all required information about the system. 5. Return to the Active system screen. Here the Results screen and the Realtime screen can be opened for viewing the incoming results and peaks. 6. In the meantime a new table can be prepared in the macro for the next run. 7. Wait until the message Sampling completed, user can change tray is displayed (see Figure 8.25) in the task bar, exchange the sample trays from the sampler for the trays for which the new table was created or place the new samples in the trays according the new table.

163 Chapter 8 The Analysis run Wait until the message Allowed to start the overlapping run is displayed in the task bar. During this stage most menu items are not available (see Figure 8.26). 9. Return to the macro, wait a few seconds and then start the run from the macro. Note: When the Realtime screen was still open from the first run, the start of the new run is already displayed while the previous run was not yet closed. Then, when the first run is finished, the Realtime screen removes the displayed peaks of this run and continued with the peaks of the new run. The same counts for the results in the Results screen. The run is packed and saved and can be opened from the DataProcessing screen (see Figure 8.27). 10. At the end of the last run of the day, ignore the messages and wait until the run is completely finished, packed and saved.

164 FlowAccess V Stopping the analysis during the run If, for any reason, the analysis of all channels has to be stopped, click the Stop button on the toolbar. Then uncheck the group. Then for each channel is asked if the channel has to be stopped. Click Yes to stop. Warning: A SA1070 sampler cannot be interrupted in this way during the pre-dilution procedure of the run. For the procedure for the SA1070 sampler see Appendix. It is also possible to stop just one channel. Click the Stop button and then uncheck the channel to be stopped. The other channels keep running. Close the program and start up again, if another analysis is to be performed. If, for any reason the system hangs or shows the message Analysis still going on, (see Figure 8.28) and cannot be stopped by the Stop button, press Alt-Shift-F12 to force a program exit (see Figure 8.29). Figure 8.28 Press OK. The Active system is closed. Close the main screen as well and then start FlowAccess TM again. Figure 8.29

165 Chapter 8 The Analysis run Restoring a crashed run file If, for any reason the PC crashes during the run, the run is aborted. It is possible to retrieve the run file until the moment the PC crashed. Procedure: 1. Re-open FlowAccess V3 TM program and open the system of which the run was aborted. FlowAccess V3 TM will show the following message (see Figure 8.30). Figure Press the Yes button. FlowAccess V3 TM takes the DB backup. The Backup FlowccessV3 Database screen will pop up. 3. Select the storage path for the DB backup (see Figure 8.31). Place the DB backup in another directory than the original DB. Figure Press OK. The backup DB will be saved as FlowAccessV3.bak (see Figure 8.32). Make sure to select the file type All files (*.*), to be able to see the.bak file. Figure Now the file can be imported. It will be imported as a run file.

166 FlowAccess V Select Import runs in the Dataprocessing tab. The Import from Database Format screen is displayed (see Figure 8.33). 7. Select the Browse button behind the Run File name to open the Open dialog box and select the backup DB from the directory where it was stored (see Figure 8.32). 8. Type a new name for the run file (as shown in Figure 8.33) or remove the aborted run file from the directory where it was stored and then import the run with the same name. Figure Select the path for the run file (usually D or C:\FlowaccessPost\Data) and press Add to list. 10. Click Import Runs. When the file is imported successfully, this will be displayed in the screen (see Figure 8.33). 11. Close the screen. The run will be displayed in the right pane and can be opened normally. The Results and the Realtime can be viewed until the moment of the crash End of the run When the run is finished, the Real time screen and the Results screen are closed and the run is saved in the database. You are returned to the Active System screen. The status of the system changes from Started to Stopped. Warning: Do not touch the samples in the sampler until the status of the system has changed to Stopped. When the post-dilution option was selected (see chapter and 6.3.5), the system will automatically perform post-dilutions and analyze them. From the time the last sample of the normal run is taken, until the performance of the post-dilution, the sampler is off. The status of the system however stays on Started and the Real time screen and the

167 Chapter 8 The Analysis run 8-29 Results screen stay open. The results of the post-dilution analysis are automatically added to the results table. Also the sample table is adapted with the post-diluted samples. Attention: If at the end of the run the error message Error in restoring runs. Please copy the run files and restore manually appears on screen the run can still be saved. Follow the procedure described in chapter When the status of the system is Stopped, the results of the performed analysis cannot be viewed in the Active System anymore. Now either the run file can be viewed in PostAnalysis or a new analysis run can be started. When another analysis is to be performed: 1. Click the Pointer button. The system is active again. 2. Check if you want to perform an analysis on the same methods or ranges and groups. If not, remove the method and/or group from the sampler and load another method from the list of available methods and /or create a new group. 3. Click the Table icon and then on the sampler. Edit the current table for the new run or remove the table and either create a new table or load an existing table. 4. Select the Baseline button and check the group to be analyzed. 5. Click the Start button 6. Open the Real time screen to check the baseline. 7. If the baseline is OK, close the screen. 8. Click the Stop button and uncheck the group. Click Yes for each channel. 9. Select the Analysis button and check the group to be analyzed. 10. Click the Start button to start the analysis. To open the run file in PostAnalysis: 1. Close the Active System screen. You are returned to the Main screen. 2. Select the DataProcessing tab. The run files are then displayed in the right part of the screen 3. Select the required run file name from the list and click on the PostAnalysis button in the left part of the screen or double click on the run file name. The system with the configuration of the run is displayed in the FlowAccessV3Post screen. The FlowAccessV3Post screen is explained in Chapter 9 Data Processing Storing the run files manually When, at the end of the run, the error message Error in restoring runs. Please copy the run files and restore manually appears, the following procedure must be carried out: 1. Select the Data processing tab. The run file is placed there with the notification behind the file (see Figure 8.34) is displayed.

168 FlowAccess V Figure Select Import runs (see Figure 8.35). Figure Select the Browse button (folder icon) behind the Run File name to open the Open dialog box (see Figure 8.36) and change the directory to the directory where your run files are normally stored (usually C or D:\FlowAccessPostV3/Data). 4. Select All for file type.

169 Chapter 8 The Analysis run 8-31 Figure Now select the file from the list that shows no extension and select Open. The dialog box is closed and the file name is displayed as file to be imported (see Figure 8.37). Figure Change the Run Display name and the Database Reference name by e.g. adding MR (= Manual Restore). 7. Select Add to list. The file is now displayed in the lower field. 8. Select Import Runs. After a few seconds the screen will show that the file was imported successfully. The dialog box is closed and the file will now be visible in the Data processing screen. Note: Preferably solve the problem before the next analysis is run. The problem may be caused due to a system privileges error. Make sure to have full Administrator rights on the PC and enough disk space.

170

171 FlowAccess V3 TM CHAPTER 9: DATA PROCESSING Preface: This chapter gives detailed instructions about editing and calculation of the results and the presentation in reports

172

173 Chapter 9 Data Processing DATA PROCESSING 9.1 Introduction As soon as the run is finished and the data is saved in the database, the run can be opened in Data processing. In the Data Processing program the measured data are calculated and the results are displayed. The results can be viewed, edited, recalculated and printed. The results can be displayed in a flexible report for printing (optional). The results can be edited by means of peak editing, but also by means of editing the method or the sample table. The Data Processing module is opened from the main screen in the FlowAccess program. Select the Data Processing tab in the left pane to activate this part of the program. The run files are displayed in the right pane (see Figure 9.1). Figure 9.1 In the upper left part four buttons are displayed: PostAnalysis, Import runs, Export runs and Analysis Run info. Also two tabs are present: Options and Run groups. When the Options tab is active all run files are displayed in the right part of the screen; when the tab Run groups is active the run files are divided in groups and only the run files of the selected group is displayed (only when run groups were made).

174 FlowAccess V Importing runs Run files from other systems can be imported. To import a run file: 1. Click on the Import Run button. The Import from Database format screen is displayed. 2. Select the Browse button and move to the directory where the run file is located. 3. Select the run file and click Open. 4. Select the run file storage path by means of the Browse button. 5. Select the Run group where the file must be placed from the drop down list and click Add to list. The file is then shown in the list to be imported (see Figure 9.2). Figure 9.2 Import runs 6. Follow the same procedure for each run file to be imported. 7. When all required runs are displayed, select Import Runs. The runs are imported into the chosen Run group(s). 8. Click Exit to return to the Main screen. 9. If required, update the run files to the current database version as described in chapter A new feature is added to FlowAccess TM for easy import of multiple runs. 1. To import multiple runs: 2. Select In Explorer the directory where the files to be imported are stored. 3. Resize the Explorer window to a smaller size, thus showing in front of the DataProcessing screen of Figure Select the run files and drag the files to the DataProcessing screen. The files will displayed in this screen.

175 Chapter 9 Data Processing Exporting runs Run files from this system can be exported. To export a file: 1. Select the file to be exported from the run file list. 2. Click on the Export runs button. The Export to Database format screen is displayed with the selected file as run type (see Figure 9.3). 3. Select the Browse button and move to the directory to where the run file must exported. 4. If all existing runs are to be exported, check this option. When the selected run can be deleted after exportation, check this option. 5. Click Confirm. The file is exported and when ready a message is displayed. 6. Click Exit to return to the Main screen. Figure 9.3 Export runs Remark: Run files can optionally also be exported via a macro to an external application as LIMS from the PostAnalysis screen. For this purpose the option Execute the external application is checked in Preferences, General settings (see chapter ) and the external application is defined. The macro can then be started from the File menu in PostAnalysis (see chapter 9.2). This option must be user defined and is only available on request Analysis Run info Info about the displayed runs can be viewed without opening the system. Select the run file of which information is to be displayed and click Analysis Run Info in the left pane. The info screen will be displayed (see Figure 9.4) with the name of the system.

176 FlowAccess V3 9-4 Figure 9.4 From this screen the RealTime (Chapter 9.3) and Results screen (Chapter 9.4) can be opened and the run can be recalculated and restored here as well. Another way to open the RealTime or Results screen, as well as the info screen is to select the Run file and right click the mouse button to display a menu from which the items can be selected directly (see Figure 9.5). Figure 9.5

177 Chapter 9 Data Processing Comments Comments can be added to the run files. 1. Select a run. Select PostAnalysis from the File menu and then Comments. The User Comments screen is displayed (see Figure 9.6) with the current information. Figure 9.6 Comments 2. Type the comments in the New Comments field and click OK. The screen is closed and you are returned to the Main screen. 3. If desired Run comments can be added as well by clicking the Change button and typing the comment in the popup field. The new comments are added to the original comments with date and time and the name of the person that was logged on. The Run comment can be deleted afterwards, however the date and time of the change, as well as the name of the person, cannot be deleted. These comments are not visible in the Main screen, but can be viewed by using the User Comments option again. By using the Back and Next button one can toggle easily between the run files to view the comments. Note: The Comments screen can also be opened by selecting a run, right clicking the mouse button and then selecting Comments from the menu Run groups Run files can be organized as desired by using run groups. Each group can stand for a special type of analysis. Run files can be retrieved more easily as usually the file list is shorter when the files are divided in different groups. Select the Run Groups button. Click inside the field with the right mouse button. Four options will pop-up: New groups, Delete groups, Rename and Refresh (see Figure 9.7).

178 FlowAccess V3 9-6 To set up file groups: 1. Select New groups. 2. A dialog box is displayed where a name must be typed for the new group. 3. Click OK. The dialog box is closed and the new group is added. Figure 9.7 File groups To delete file groups: 1. Select the group name to be deleted. 2. Select Delete groups. The group and all the files belonging to that group are deleted. To rename file groups: 1. Select the group to be renamed. 2. Select Rename groups. A dialog box is displayed where the new name for the group is typed. 3. Click OK. To move between file groups: 1. Select the file to be moved. 2. Click the right mouse button to display the menu (see Figure 9.5). 3. Select the last option, Change Run groups (see Figure 9.8). The groups that are present are shown. 4. Click with the left mouse button on the group where the file is to be to be displayed. The file is moved to the selected group. Figure 9.8

179 Chapter 9 Data Processing FlowAccessV3Post Double click on the run to be opened or select the run and click the PostAnalysis button in the left part. The FlowAccessV3Post screen is displayed with the system configuration of the run (see Figure 9.9). Figure 9.9 PostAnalysis screen The screen looks like the Analysis screen, although some menus and buttons are not displayed, as they are not relevant for PostAnalysis. The filename of the run is displayed in the title bar. The File menu shows the options Restore run - Previous version (to be used when 21CFR part 11 is applied), Restore run - Original version (to be used for other analyses) and Execute the external application. The first two options are to be used to restore the analysis when e.g. peaks were edited. The third option is only available when this option was checked in Preferences, General settings, see chapter and is used for exportation of run files to an external application. This macro must be user defined and is only available on request. An example for this macro is shown in Figure Figure 9.10

180 FlowAccess V3 9-8 The Analysis menu shows the extra options Data Evaluation (see chapter 9.2.1) and Master/Slave Settings. The latter is described in chapter and the screen shows the settings as set for the analysis, which can be changed. One new item is available in the View menu: Run History, where info about the run can be viewed. The Remote Control button and the Groups button can be used to display the settings for sampler respectively the analyzer. These settings cannot be changed. The table can be edited by clicking the icon (see chapter ). The table is displayed as before the run was performed (no results displayed, as the results cannot be edited in the table). The method can be edited for calculation by clicking the icon (see chapter ) and then on the method. For the methods only those options that are relevant for the calculation of the method can be edited. All options relevant for the performance of the run are not available. When a sample type is changed, the results are immediately recalculated in the Results screen. Note: Table and method editing is also accessible from the Results screen. The Real time screen can be opened to display the complete run. The peaks can be edited in this screen and the start value changed (see chapter 9.3). The Results screen shows the results, calibration curve and statistics of the run, while method and table can be edited (see chapter 9.4). In the Results screen the results can be viewed, recalculated and printed Data Evaluation Data Evaluation shows the following screen (see Figure 9.11). Figure 9.11 Data Evaluation In this screen the damping for each method can be changed as well as the matrix settings, after which the results are automatically recalculated (before the damping is changed the Results screen must be closed). The option Apply Savitzky-Gohlay is used for the normal detection channels. The options for the matrix settings are then not available. For each available channel the damping can be set and changed. The option Matrix settings is used for matrix detection channels. The factor and Offset settings for the matrix channels can be changed. To change or apply damping, first the filter must be checked.

181 Chapter 9 Data Processing Real time screen To open the Real time screen to display the complete run, click the Real time button on the toolbar or select Real time from the Analysis menu. The Real time screen is displayed by default in the tiled view with one channel active (see Figure 9.12). Figure 9.12 Realtime all channels Click in the field of any channel to make the channel the active channel. The active channel can be displayed in full window by double clicking in the title bar of the channel or clicking the Maximize button. As soon as a channel is displayed in full window a toolbar becomes available with viewing and editing options (see Figure 9.13). Figure 9.13 Realtime one channel

182 FlowAccess V Two extra menus are available: Format for all display aspects and Tools for all information and editing purposes. All items of the Format and Tools menu are also available from the toolbar (except for the Pause and Play options, which are only available during the run). By default an X-Y guide is displayed. Other display settings, as grid, peak labels and measurement windows can be selected. These display settings are saved when the screen is closed and will be shown when the screen is re-opened. By means of the Channels menu can be returned to the view with all channels or can be toggled between the channels. By means of the Views menu two other views can be displayed: Overlayered view and Multi scale view (see chapter 9.3.5) Peak information and editing The measuring points in the screen can be viewed and edited. For example if a peak is spiked due to an extra air bubble. Warning: Peak editing is not allowed in PostAnalysis while another analysis is running, as the recalculation after editing interferes with the current run. Note: Peak editing can only be performed when the X-Y guide is displayed. A message will be displayed to select the X-Y guide as soon as a peak is selected. Peak editing is performed in the Peak information dialog box, which can be opened from the Tools menu, by clicking the icon in the toolbar or by double clicking on a peak. Information about each separate peak can be displayed. By default information about the first peak of the screen is displayed, when the dialog box is opened from the Tools menu. When clicking the Forward button the information of the next peak is displayed. In the Realtime screen the peak of which information is displayed is circled (see Figure 9.15). For easy move to a certain peak, without closing the Peak information box the peak number can be entered and after pressing Go the required peak will be displayed Figure 9.14

183 Chapter 9 Data Processing 9-11 When double clicking on a peak this peak is displayed directly in the Peak Information box or just double clicking inside the peak window displays the Peak information box with the nearest peak selected. The view of the Peak Information box depends on the selected peak detection method(s). When multiple peak detection was selected the view of Figure 9.15 is displayed. When single peak detection (min/max. approximation) was applied the view of the Peak Information box is different (see Figure 9.16). Figure 9.15 Peak information box for Multiple peak detection Figure 9.16 Peak information box for single (Min/max) peak detection Peak editing for both peak detection methods is somewhat different Multiple peak detection The peaks are always calculated as set in the Method settings (Automatic Peak Detection). When one (or more than one) detection method is deselected from calculation, this is shown by the peak flag, which will show a [W] (warning) or an [E] (Error). This can be due to setting a very low tolerance. Check the tolerance in the Method settings if it is 2.5% of the highest standard. If not, change this, save the method and allow new peak detection. Mostly all peak detection methods are then used and the warning or error disappeared. When peak labels are shown, the color of the peak label indicates whether the calculation of the peak is ok or not. Black = ok, brown = Warning, red (striked through) = Error Peaks, that are used for calibration (standard) or correction (drift, wash) can be deselected from calculation. The peak labels show an {X} It is possible to change a peak detection to semi-automatic peak detection or manual peak detection.

184 FlowAccess V Semi-Automatic Peak detection 1. Select the option. All detection methods become available. 2. Each detection method can be deselected. The marker of the method on the peak disappears, when deselected. 3. Press Save to save the new peak detection. The box is closed. 4. When the box is re-opened, the recalculated result is displayed. A flag [W] or [E] is placed for the peak, when detection methods were deselected. These flags are also visible for the result in the Results screen (when flags are selected for the view) after re-calculation. Peak labels get a brown color when one peak detection was deselected and a red color (and striked through) when more than one was deselected. Manual Peak Detection 1. Select the option. 2. Double click with the cross-hair on the spot where the peak optimum is assumed. The peak is marked. 3. Press Save to save the new peak detection. The box is closed. 4. When the box is re-opened, the recalculated result is displayed. All official detection methods are deselected and the result is flagged with a [M] after re-calculation. Peak labels get a blue color and {Ed} is added to the label. In the case that the peak is over range the peak label is red (and striked through) with {Ed} added to the label Single peak detection In this case the min/max peak detection method is applied as set in the Method settings. To be able to edit the peaks a Modify button is available and a Delta option to set the steps when the arrow buttons are used to move the measuring point of the peak. 1. Double click on the peak to be edited or enter the peak number in the field GoTo Peak and then click Modify. (The button will now display Save, see Figure 9.17). Now the steps are not from peak to peak, but within a peak. Figure 9.17 Peak editing

185 Chapter 9 Data Processing Move with the Backward and Forward buttons to the left or the right to move the measuring point. The steps can be varied by the delta (1= 1 second, 2 = 2 seconds, 4 = 4 seconds etc.). 3. Click Save when the measuring point is on the proper place. The setting is saved and the Peak information box is closed. The measuring point is placed on the new place (see Figure 9.18). The peak label will be blue and {Ed} is added to the label. Figure 9.18 Edited peak 4. When the peak information box is opened again by double clicking the peak, the modified peak height and measuring time of the edited peak are displayed. The text Peak edited by user is displayed. 5. With the Forward button can now be moved to another peak to be edited. 6. The results are automatically recalculated in the Results screen.

186 FlowAccess V When in long runs peaks are not picked correctly, or when as shown above, a spike was picked, it is possible to correct the peak picking by selecting the option Apply this window time (see Figure 9.19). Figure 9.19 Peak correction First edit the peak to the correct position, then select Apply this window time and click Save. Then all peak picking after the corrected peak is shifted (see Figure 9.20). Figure 9.20 Corrected peaks Note: When peaks are edited in the Realtime screen, the results will be different for that method. These new values are not updated directly in the Results screen. Select Recalculate from the File menu in the Results screen to view the new values.

187 Chapter 9 Data Processing Peak picking parameters Select Peak Picking parameters from the toolbar or Tools menu Break information Figure 9.21 Peak parameters The Edit peak picking values dialog box (see Figure 9.21) is opened, where the settings as set in the Method Settings are displayed. Editing of the settings is only valid for the selected method. Note: Editing of the peak picking values can also be performed from the Results screen in Edit method. The damping on the peaks can be changed here as well. The Apply button under the option has to be pressed for applying the new damping option for that method. Note: The results are not updated directly in the Results screen. Select Recalculate from the File menu in the Results screen to view the new values. The damping can also be changed in the PostAnalysis screen by selecting Data Evaluation from the Analysis menu. In the case of a run with post-dilutions the Break information can be retrieved from the tools menu. An example is shown in Figure When a run has ended with over range samples a cross is placed on the run-out wash. The samples are post-diluted and when the run is restarted a new cross is placed to indicated the start of the post-dilution run. The system checks in this way the time from the start until the first peak. The post-dilution run starts with a drift/wash combination. In this example run a post-dilution run and then 2 cascade dilutions had to be run. This means that after the original run stopped a break was held until the samples were diluted. The analysis started again and stopped when all dilution positions were run. As some samples were still over range a break was held for preparing post-dilutions and started again. This was repeated once more until all sample results were within the measuring range.

188 FlowAccess V Peak labels Figure 9.22 Break information The view in Realtime can be adapted by showing peak labels. This option can be opened from the Tools menu or the toolbar (see Figure 9.23). A selection can be made between None, Identity, Results and Identity and Results. Figure 9.23 Peak labels When the run is printed from the Realtime screen, the labels are printed as displayed in the window.

189 Chapter 9 Data Processing More Views Select More Views from the Views menu. The Overlay display mode view is displayed (see Figure 9.24). Figure 9.24 In this screen the channels can be displayed overlaid. Select the channels to be displayed by checking the tick boxes in the left part of the screen. An example with 4 channels displayed is shown in Figure The view can be zoomed in and zoomed out. Figure 9.25 Overlayered view

190 FlowAccess V A Multi Scale view can be shown with selected channels when clicking the button in the left part of the window (see Figure 9.26). Figure 9.26 Multi scale view In this view cannot be zoomed. 9.4 Results screen Select Results from the toolbar or select Results from the Analysis menu. The Results screen is displayed with the results of the analysis in the default view. The screen is displayed with one menu: Options and four toolbar buttons: Format, Multi Pickups, Print and Exit. Depending on the analysis the screen is displayed with four or five tabs: Detailed results, Cumulative results (only when pre-dilutions and/or postdilutions have been performed), Calibration, Statistics and Edit Table & Method. Not all menu functions can be used in all tabs Options menu The functions of the Options menu are: Format Results, Re-calculation, Print, Export, Duplicate view, Sheet format and Calibration Constants. Format Results The view can be changed by using Format Results from the Options menu or by selecting the Format button on the toolbar. This option is described in chapter Re-calculation This option must be used after editing results, either in the Realtime screen as in the Results screen to achieve correct results. Print The results can be printed by using Print from the Options menu or by selecting the Print button on the toolbar. First a Print screen is displayed where the printer and the print properties can be selected. When the settings are accepted the Print Preview screen is displayed where the preview of the print is shown (see Figure 9.27).

191 Chapter 9 Data Processing 9-19 Figure 9.27 Print preview results When the printer icon is selected the file is printed. Export The results can be exported to either Excel or ASCII format. When selected the Save as dialog box is displayed where the file must be given a name with a.xls (Excel) extension or.txt (ASCII) extension. Select the directory where the exported file is to be saved. The.xls file can only be opened when Excel is installed on the PC. An example of a.txt file and a.xls file are shown in Figure 9.28 and Figure Figure 9.28 File in Notepad

192 FlowAccess V By means of the Sheet Format it is possible to select only certain columns and sample types for export. See for this option further in this chapter. Figure 9.29 File in Excel When result are not to be excluded from export, this can be achieved by deselecting the sample in the original sample table, see chapter Duplicate view With this option a tab is added to the views (at the bottom of the screen) where a compact view (only sample type, sample identity and the results of the channels) is displayed and, in the case that samples with the same identity were present in the table, the average result is displayed (see Figure 9.30). Figure 9.30 Duplicate view

193 Chapter 9 Data Processing 9-21 Calibration constants With this option the calibration constants of each channel can be displayed (see Figure 9.31). With the View Report option a preview of a report is displayed, which can be printed by using the printer icon. Each channel is printed separately. Figure 9.31 Calibration constants It is also possible to print the constants of all channels of a sampler together in one file. To achieve this select the sampler instead of the channel. Sheet Format With this option a customized view of your results can be created (see Figure 9.32 for an example) for export. In the Results screen the default columns of the run are displayed. This view can be used for example for a quick overview of certain results or for the export of the results to Excel or ASCII format. Create a custom view by means of the Format Results option (see chapter 8.5 for explanation) and set this view as one of the default views. Display the Results screen and select the custom tab (see Figure 9.32).

194 FlowAccess V Figure 9.32 Results with custom tab Then open the Sheet Format screen (see Figure 9.33). Figure 9.33 Customized headers can be shown with text or numbers and columns can be hidden as required. Also a selection of the sample types that will be viewed can be selected. New rows can be added where a simple calculation formula can be entered. For example: For the export of the results only the results of unknown are required, some columns are not required, a customized header is added as well as a column for a calculated result. 1. Deselect the columns that are not required. 2. Type your customized header in the Header 1 column. 3. Check Header 1 under Settings. 4. Check Unknowns under Settings. 5. Click Add to add a column to the sheet and type the next column number, the headers, check it to be visible and type the formula for the column. In this case the result (in four digits) of column 8 minus column 6 must be shown.

195 Chapter 9 Data Processing The sheet will look like Figure Click Save. Figure The Sheet format is closed and the created view is displayed (see Figure Figure 9.35 This custom tab as defined with the Sheet format will be displayed by default every time the Results screen is opened. The tab Default view stays as it was in the original view Detailed results In Figure 9.36 an example of the Detailed results of a run with pre-dilutions and post-dilutions in the Default view.. For the cup position the cup number is displayed where the sample is taken. In the column PDcupshistory the original sample cup for pre-diluted and post-diluted samples is displayed. All menu and toolbar functions of the screen can be used in this view.

196 FlowAccess V Figure 9.36 Detailed results As explained in chapter the peak information on each result can be displayed by double clicking on the result. Editing in the Peak information is explained in chapter Cumulative results When this run is viewed in Cumulative Results (see Figure 9.37) only the final recalculated results of the samples are displayed. Figure 9.37 Cumulative results For the cup position the cup number is displayed where the last dilution is taken. In the column PDcupshistory only the original sample cup for prediluted samples is displayed. In this view the menu functions cannot be used and Peak information cannot be opened here.

197 Chapter 9 Data Processing Calibration In the Calibration tab the calibration curves of the methods are displayed (see Figure 9.38). Figure 9.38 Calibration curves The calibration type can be changed after clicking the down arrow and selecting another calibration type from the list. Only the calibration types according ISO are displayed with confidence intervals. The calibration constants are displayed below the curve. Up to four calibration curves can be displayed in one view. If more calibration curves are present these curves can be viewed by clicking the Next button. The calibration curves can be printed. Select a curve and then click the Print button. Each curve has to be printed separately. A standard can be removed from the calibration curve. Refer to chapter for editing the calibration curve The calibration constants can be viewed separately and printed (refer to chapter

198 FlowAccess V Statistics In the Statistics tab the statistics of the run can be viewed (see Figure 9.39). Figure 9.39 Statistics screen The statistics for the washes, the drifts, the unknowns (and typed unknowns, QC s, CLP s and Spikes can be displayed for the selected channel (by default the first channel of the result file is selected). Up to four types can be displayed simultaneously. If an alarm limit (for unknowns or QC s) was entered in the method, these limits will be displayed. The scale of each pane can be changed and each pane can be printed separately. Each pane can be exported separately in a Tee Chart format. By default the upper left pane is the active pane. Any pane can be made the active pane by clicking in the pane. To display the statistics: 1. Select the required channel in the toolbar. 2. Click in the pane where the graph is to be displayed to make it active and then click on the graph type to be displayed. 3. By default the limits are from -1.0 to 1.0 (see upper left graph of Q1 in Figure 9.40). To zoom in drag with the mouse from upper left to right bottom over the displayed figures (see upper right graph of Q1 in Figure 9.40). 4. Repeat step 2 for the other panes, if required. The graphs will displayed in the panes (see Figure 9.40) for an example with graphs for QC s). Note: When the values are within the set limits the color is green, when outside the limits the color is red.

199 Chapter 9 Data Processing 9-27 Below the pane information is displayed about the graph type (total number, max. result, min. result and average result. Figure 9.40 Statistics of QC s The view of each pane can be changed by means of the tools in the toolbar (home, end, zoom). When Viewinfo is clicked the x-value and y-value of the mouse in the pane are displayed in the toolbar. The view can be refreshed by pressing Refresh. To print a graph: 1. Click in the graph to select it for printing. 2. Select Print Preview from the toolbar. The Print Preview screen is displayed (see Figure 9.41). Figure 9.41 Print preview

200 FlowAccess V Select the printer on which the graph is to be printed. 4. Select the required orientation and the margins of the paper. 5. Press Print. The graph is printed. 6. Press Close to close the screen. The graphs can be exported in a graphic format. To export a graph: 1. Click in the graph to select it for exporting. 2. Select Export from the toolbar. The TeeChart export screen is displayed (see Figure 9.42). 3. Select the export format for the graph from the displayed list. 4. If required select Figure 9.42 TeeChart the color options and the size of the exported picture. 5. Press Save. 6. Select the destination directory where the graph is to be stored and press Save. 7. Press Close to close the screen. To change the scale of a pane: 1. Select the pane of which the scale has to be adapted. 2. Select either x-value or y-value from the toolbar. A dialog box is displayed where the limits can be set. 3. Adapt the settings as required and click OK. The box is closed and the pane is displayed with the new settings Used formulas in statistics Correlation Coefficient. The relation between the corrected peak height and the theoretical concentration of the standards r = xi. yi xy i i n ( xi) 2 ( yi) xi yi n n Symbols: x i = corrected peak height of the standard i y i = theoretical concentration of the standard i n = number of standards The more r is closer to , the better results correlate Note: This coefficient is not changed or affected by use of 2 nd order.

201 Chapter 9 Data Processing 9-29 Remark: Justification of using an amount of samples depends on the required and statistically proved working range of the analytical method. The interval, being experimentally established and statistically proved by the calibration of the method, lies between the lowest and highest quantity or mass concentration. The lowest possible limit of a working range is the limit of detection of an analytical method. Baseline Correction. The baseline correction function is required for peak height calculation and corrects for the reagent-offset level (see Figure 9.43). Baseline correction is the first correction function used and precedes always the peak height sensitivity drift correction. This normally constant baseline level might change over time, due to various circumstances as change in reagent quality, temperature, etc. Therefore at periods of time new correction measurements are made and recalculated in results. Formula: w( b) w( a) U' = U - ( n- a). w b- a ( n) ( n) ( a) Symbols: U (n) = Baseline Corrected Peak Height for sample cup n. U (n) = Measured Uncorrected peak height for sample cup n. n = Sample cup number. W (a) = Measured peak height of preceding wash cup. W (b) = Measured peak height of following wash cup. a = Cup number preceding wash. b = Cup number following wash. Note: Initial Wash is the first wash position, automatically taken at the start of the run. Figure 9.43 Baseline correction

202 FlowAccess V Sensitivity Correction. The Sensitivity Correction function is required for peak height calculation and corrects for sensitivity change over time periods during the analysis run (see Figure 9.44). The value is always corrected for baseline drift. Normally this function corrects for a minimum change, but is still necessary as chemical reactions are influenced by changes in temperature, reagent conditions, etc. Therefore at periods of time new correction measurements are made and recalculated in results. Formula: D1 U ''( n) = U '( n) D(b) - D(a). (n - a)+ D(a) b-a Symbols: U (n) = Baseline and Peak Drift corrected peak height for sample cup n. U (n) = Baseline Corrected Peak Height for sample cup n. D1 = Baseline Corrected Peak Height of the first Drift cup. D(a) = Baseline Corrected Peak Height of the preceding Drift cup. D(b) = Baseline Corrected Peak Height of the following Drift cup. n = Sample cup number. a = Cup number of preceding Drift. b = Cup number of following Drift. Note: A drift should never be used as a quality control as the value for the drift itself, displayed in the results, is only corrected for baseline drift and not corrected for sensitivity drift. Figure 9.44 Sensitivity correction

203 Chapter 9 Data Processing 9-31 Example of calculations of corrections (16-bit detector) Columns displayed by the PC Position Type1 Identity1 Result measured height corrected height Range Baseline corrected height Sensitivity corrected height WT IW Initial Wash 0, ST2 T Tracer 0, Wt W Wash 0, SP1 S1 sample 0.1/0.02 0, SP2 S2 sample 0.2/0.04 0, ST2 S3 sample 0.5/0.1 0, SP3 S4 sample 1.0/0.2 0, SP4 S5 sample 2.0/0.4 0, SP5 S6 sample 5.0/1.0 0, ST1 I Sample 10.0/2.0 1, Wt WI Wash Ignore 0, ST2 D Drift 0, D1=873 Wt W Wash 0, A1 U Unknown-1 0, A2 U Unknown-2 1, A3 U Unknown-3 0, A4 U Unknown-4 1, A5 U Unknown-5 0, A6 U Unknown-6 0, A7 U Unknown-7 0, A8 U Unknown-8 0, A9 U Unknown-9 0, A10 U Unknown-10 0, Wt WI Wash Ignore 0, ST2 D Drift 0, a=24 D(a)=879 Wt W Wash 0, a=25 W(a)= B1 U lot-1 1, B2 U lot-2 1, B3 U lot-3 1, B4 U lot-4 1, n=29 U'(n)=29794-[(29-25)*( )/(37-25)] = B5 U lot-5 1, B6 U lot-6 1, B7 U lot-7 1, B8 U lot-8 1, B9 U lot-9 1, Wt WI Wash Ignore 0, n= 29 U'(n)= U"(n)=15366*(873/{[( )*(29-24)/(36-24)]+879}) =14826 ST2 D Drift 0, b=36 D(b)=941 Wt W Wash 0, b=37 W(b)=14483 Wt W End Run 0, Wt E End Run 0,

204 FlowAccess V Calculation of Calibration Constants To calculate results standard solutions with known concentrations are analyzed. It s measured, and corrected, peak height is used to prepare a calibration curve from which unknown concentrations can be calculated. This calibration constants can be obtained in different ways: 1 st and 2 nd order, according ISO 8466, 1 st and 2 nd order including origin; the amount of standards in the working range (minimal 2 standards for 1 st order and 3 for 2 nd order calculations). Calculation 1st order, according ISO 8466, linear calibration curve Formula #1. y = a + bx Symbols: y = Baseline corrected and peak drift corrected Peak Height of the Standard. x = Concentration of the Standard. a = Ordinate of the intercept. b = Sensitivity of the method (slope of the calibration line; coefficient of regression) The calculated values a and b will be used for Calculation of Results of Unknown Samples. Ad A. Calculation of the sensitivity of the method is obtained from above formula #1 via Formula #1A b = N i = 1 ( x X).( y Y) i N i = 1 ( x X) i i 2 Symbols: N = Number of measurements. x = Concentration of the standard. i y i = Corrected peak height. X = Average of concentrations. Y = Average of corrected peak heights. Ad B. Calculation of the ordinate intercept is obtained from the above formula #1 via Formula #1B a = Y bx Symbols: b = Sensitivity of the method X = Average of concentrations Y = Average of corrected peak heights

205 Chapter 9 Data Processing 9-33 Calculation 2 nd order according ISO 8466, non-linear calibration curve Formula #2: y = a + bx + cx Symbols: y = Baseline corrected and Peak Drift corrected Peak Height of the Standard. x = Concentration of the Standard. a = Ordinate intercept. b,c* = Sensitivity of the method (slope of the calibration line; coefficient of regression) d d = + (slope at x 0) x * b 2cx0 t The calculated values a, b and c will be used for Calculation of Results of Unknown Samples. (Basic detailed formulas): 2 a. Q xx x 2 i = xi N 2 b. y i Qxy = ( xy i i ) xi. N c. 2 x 3 i Q 3 = xi x. x i N d. Q x 4 = x xi N 4 i 2 2 e. Symbols: N = Number of measurements. x = Concentration of the standard. i y i = Corrected peak height. 2 x 2 i Q 2 = ( xi. yi) yi. x y N

206 FlowAccess V (Center of the working range) x x = i N (Average information value) y y = i N Ad A. Calculation of c is obtained from the above formula #2 via Formula c = ( Q. Q )-( Q. Q ) xy 3 2 x x y xx 2 ( Q 3) -( Qxx. Q 4) x x Ad B. Calculation of b, the sensitivity of the method, is obtained from the above formulas via Formula Q b = cq xy x 3 Q xx Ad C. Calculation of a the ordinate intercept is obtained from the above formula #1 via Formula a y b x c x 2 i i i = N Calculation 1 st order forced through the origin. As described at Formula #1, but in this calculation a is fixed at 0 (zero) y = 0 + bx Calculation 2 nd order forced through the origin. As described at Formula #2, but in this calculation a is fixed at 0 (zero) y = 0 + bx + cx 2

207 Chapter 9 Data Processing 9-35 Calculation of Results of Samples. In general, the final result for a sample is calculated according the formula: Result = Raw result * total dilution / weight Raw result = result obtained from the calibration curve using the peak height of the sample, corrected for baseline and sensitivity drift (this raw result is not displayed) Total dilution = External dilution * Pre-dilution * Post-dilution Weight = Weight factor entered in the table under the heading Weight With a 1 st order Linear Calibration Function. y x a b y = a + bx = Baseline corrected and peak drift corrected peak height of the sample. = Result of the analyzed sample. = Ordinate of the calibration straight line (calculated blank) = Sensitivity of the method (slope of the calibration line; coefficient of regression) With a 2 nd order Non-Linear Calibration Function. y = a + bx + cx y = Baseline corrected and peak drift corrected peak height of the sample. x = Result of the analyzed sample. a = Ordinate of the calibration straight line (calculated blank) b, c = Sensitivity of the method (slope of the calibration line; coefficient of regression) With a 1 st order, forced through origin. y = a + bx y = Baseline corrected and peak drift corrected peak height of the sample. x = Result of the analyzed sample. a = 0 b = Sensitivity of the method (slope of the calibration line; coefficient of regression) With a 2 nd order, forced through origin. y = a + bx + cx y = Baseline corrected and peak drift corrected peak height of the sample. x = Result of the analyzed sample. a = 0 b, c = Sensitivity of the method (slope of the calibration line; coefficient of regression) 2 2

208 FlowAccess V With a 3 rd order Non-linear Calibration function 2 y = a + bx + cx + y = Baseline corrected and peak drift corrected peak height of the sample. x = Result of the analyzed sample. a = Ordinate of the calibration straight line (calculated blank) b, c,d = Sensitivity of the method (slope of the calibration line; coefficient of regression) dx 3 With a 1 st order, inverse logarithm y x a b y = bln( x) + a = Baseline corrected and peak drift corrected peak height of the sample. = Result of the analyzed sample. = Ordinate of the calibration straight line (calculated blank) = Sensitivity of the method (slope of the calibration line; coefficient of regression) With a 2 nd order, forced through origin y = cln( x 2 ) + bln( x) + a y = Baseline corrected and peak drift corrected peak height of the sample. x = Result of the analyzed sample. a = Ordinate of the calibration straight line (calculated blank) b, c = Sensitivity of the method (slope of the calibration line; coefficient of regression)

209 Chapter 9 Data Processing Edit Table & Method Table editing Note: Tables and Methods can also be edited in the FlowAccessV3Post screen. By means of the Edit Table & Method tab both table and method can easily be changed. This can be convenient if for example a (combined) standard or a sample appears to be wrong. The type of the standard can be changed into an unknown in the table. Then the standard is excluded from the calibration curve for all channels. Note: To exclude the standard in one channel double click on the result to open the Peak Information box and exclude the peak from calculation or double click in the calibration curve of the channel and deselect the standard (refer to chapter 8.6.1) A sample is not really excluded from calculation, however when in the table one or more channels are deselected and the table is saved, the results of the deselected channel(s) are displayed as Such negative results are excluded from exportation to e.g. LIMS. Note: Standards cannot be excluded from calculation by deselecting one of more channels for the standard in the table. To edit a table: 1. Select a table from the Table Entry list and click Edit. The table as it was setup and analyzed is displayed (see Figure 9.45). Figure Click in the field to be changed and change as required or deselect the result of a sample in the channel (for example in the table in Figure 9.45 the type of standard3 in changed into unknown and the sample in position E60 is deselected for all channels). 3. Click Save. The table is automatically closed.

210 FlowAccess V The results are automatically recalculated and can be viewed in the other tabs of the Results screen. The Detailed Results screen (see Figure 9.46). shows Standard3 as unknown (yellow colored) and the result of the sample on position E60 is displayed as Method editing Figure 9.46 Not all parameters can be changed. The parameters that influence the performance of the run cannot be changed and are grayed out in the method. Standards can be deselected for a method and will be removed from the calibration curve after saving the method. The Peak detection method can be changed and will affect all samples of the method. To change the Peak detection method of a single sample refer to chapter and 9.3.1). Note: The damping options of the method are gray and cannot be changed here for calculation reasons. The damping can be changed either in the Peak picking parameters in the Realtime screen (see chapter 9.3.2) or in Data Evaluation in the FlowAccesPost screen (see chapter 9.2.1). To edit a method: 1. Select a method from the Method list and click Edit. The method as it was setup and used in the analysis is displayed. The items that cannot be changed are grayed out (see Error! Reference source not found.). 2. Change the method as required. 3. To change more methods, select the required method in the left pane.

211 Chapter 9 Data Processing Click Save and close the screen. Figure 9.47 will be displayed. 9.5 Reports Figure Click Yes. The results are then recalculated and can be viewed in the other tabs of the Results screen. The results, calibration curve, real time view and other information about the run can be setup in a comprehensive report in user-defined formats. To be able to create such a report the separate program FlowReport TM is necessary. This program is available as a separate option in FlowAccess. If FlowReport is installed: The program can be opened from the FlowAccess main menu or from the PostAnalysis screen. The program consists of two parts: Designer and Viewer. The designer is used to setup the layout of the different parts of the report. The viewer is used to view and print the different parts of the report. For the handling of the program refer to the separate manual provided with the program. If FlowReport is not installed: The results, the calibration curve and the real time view can be printed straight from the Results screen or the PostAnalysis screen. However the setup as shown in the preview cannot be changed. The results can also be exported (see chapter 9.1.2) and printed from the exported file.

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213 FlowAccess V3 APPENDIX

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215 Appendix APPENDIX Practical operations Some examples of problems and their solutions are outlined. A. Start Ignore time too short Problem: The Start Ignore time for a method was set on a time that the baseline was not yet steady. This caused wrong baseline correction of the peaks (see also chapter 9.2.1). Solution: 1. Open the run in PostAnalysis. 2. Open the Real Time window and select the method to be edited by double clicking the displayed method. 3. Measure the start time of the Tracer peak by means of the XY-guides. 4. Select Peak picking parameters from the Tools menu to open the dialog box. 5. Change the setting for the Start Ignore to a time at least 30 seconds before the start of the Tracer peak and click Apply. The Start Ignore triangle is replaced to the new setting. 6. Click Close to close the dialog box and close the Real time window (or, if necessary, perform the same action on another method). 7. Open the Results window to view the recalculated results. B. Start Value Tracer too high Problem: The Tracer is missed in the analysis of a method due to the fact that the Start Value Tracer was set too high for the sensitivity of the method. This caused wrong peak picking (see also chapter 9.2.1). Solution: 1. Open the run in PostAnalysis. 2. Open the Real Time window and select the method to be edited by double clicking the displayed method. 3. Select Peak picking parameters from the Tools menu to open the dialog box. 4. Change the setting for the Start Value Tracer to a value lower than the measured value of the Tracer peak and click Apply. The peaks are picked according to the changed settings.

216 FlowAccess V3 5. Click Close to close the dialog box and close the Real time window (or, if necessary, perform the same action on another method). 6. Open the Results window to view the recalculated results. C. Spiked Peaks Problem: Some peaks are wrongly detected due to spikes (see also chapter 9.2.2). Solution: 1. Open the run in PostAnalysis. 2. Open the Real Time window and select the method to be edited by double clicking the displayed method. 3. Select Peak information from the Tools menu to open the dialog box. 4. Move with the arrow to the spiked peak and click Modify. The triangle changes into an asterisk. 5. Click on the left or right arrow until the cross-hair is on the top of the real peak and click Save&Close. The asterisk changes again into triangle. 6. If necessary edit other peaks in the same way. 7. Close the Real time window (or, if necessary, perform the same action on another method). 8. Open the Results window to view the recalculated results. D. Wrong Calibration curve Problem: The calibration curve is very bad due to a wrongly prepared standard and consequently the calculations for the unknown samples are incorrect. Solution: 1. Select the Edit Table & method tab in the Results screen. 2. Select the Method to be edited from the list. The Method screen of the selected method will be displayed. 3. Select the Calibration tab and deselect the wrong standard to exclude it from the calibration. 4. Click the Save button to save the change. 5. Click the Exit button to close the screen. The results are automatically recalculated. E. Flushing SA 1070 sampler Problem: Air bubbles are present in the syringe of the diluter, while the Flush button may not be used by the Remote Control. Solution: 1. Switch off the sampler by the main switch at the back and then switch it on again. The sampler will automatically flush the syringe. 2. Repeat this procedure until all air bubbles have disappeared.

217 Appendix F. Interruption predilution with a SA 1070 sampler Problem: The analysis has to be stopped for any reason during the pre-dilution step of the run. Solution: 1. Switch off the sampler by the main switch at the back of the sampler. 2. Stop the analysis run by the Stop button in the Analysis window. 3. Uncheck the group and click Yes until all channels are stopped. 4. Close the program. 5. Switch off the interface. 6. Change the holders on the sampler manually to their proper position. 7. In the case you want to restart the analysis: Check if the reason for the stopping is solved. 8. Switch on the interface. 9. Switch on the sampler. The sampler will automatically flush the syringe. 10. Start the program and start the analysis in the usual way.

218 FlowAccess V3 Configuration for standards in SA1074 with 2, 3 or 4 needles

219 Appendix Configuration for standards in SA1070 for 2 needles

220 FlowAccess V3 G. Repairing Runtime error 91 Problem: When the communication module is closed by accident during analysis, this Runtime error shows up when FlowAccess is restarted. Then FlowAccess closes directly spontaneously, but leaves the communication module open, which must be closed manually. Repeating this action will only result in Runtime error 91 again. Reason: A sudden break in the communication does not stop the interface from sending raw data or other communication data. The data arriving at the RS232 is disturbing the regular startup by FlowAccess, resulting in this runtime error. Solution 1: 1. When the communication module is closed by accident, close FlowAccess. 2. Switch off the interface and switch off the sampler(s). 3. Then switch on the sampler(s); switch on the interface (actually a restart of all hardware/analyzer). 4. Start FlowAccess and login. 5. Open the Active system and proceed with the analysis. 6. Do not close the communication module at any circumstance while FlowAccess is open. FlowAccess will open and also close the communication module by internal command. Solution 2: 1. When the communication module is closed by accident, close FlowAccess. 2. Switch off the interface and switch off the sampler(s). 3. In FlowAccess : Open Settings, Options and select tab General. 4. Deselect the tick box Show Communication Module. 5. Press OK. 6. Close FlowAccess. 7. Restart FlowAccess. 8. Open the Active system and proceed with the analysis. Remark: In this case the communication module is not visible in the task bar at the bottom of the screen, nor in the task manager at the tab Applications ( it is only visible at Processes ). When logging is required at moments of troubleshooting, close FlowAccess and start the communication module.exe from the subdirectory in Windows explorer. Change the logging settings and close the communication module. H. Flowchart for screening On the next page an example for how to setup an analysis with screening.

221 Appendix