EAS32 v.1.4 Costech International SpA

Transcription

1 EAS32 v.1.4 User s Guide Rev. D Tel: Costech International SpA Fax: Via Firenze 30/A info@costech.net Cernusco s/nav. Milan Web: Italy I

2 II

3 Using the document The User's Manual offers the users of the chromatographic station EAS32 a detailed description of controls, a procedure for processing the first analysis, detailed guidance concerning the fundamental types of analyses and calibrations as well as copious additional information about the options and variants relevant for working with the station. Chapters 1, 2, 3 and, in particular, Chapter 4 - First Analysis - are earmarked for beginners who will quickly find the required information there. Once acquainted with the fundamental procedures the user is recommended to turn to Chapter 10 - Resolving Problems, which contains a list of problems most often encountered when working with the EAS32 station. Summary information about all commands and error messages can be found in the Reference Manual. The User's Manual presupposes prior knowledge of the fundamental concepts of the Windows and MS-DOS operating systems like file, directory, path, etc. The following fonts are used in the manual to set apart some parts of the text: Open Channel WORK1 Enter RUNNING Supervisor Commands and items - bold Window titles - bold Sans Serif filenames and directories - capitals key designations - Sans Serif font in a box possible states of the station - capitalised Italics values of parameters - Italics. With wishes of easy and lucid reading Costech III

4 IV

5 User s Guide Contents Contents 1 Introduction Description of the Elemental Analysis Software Updating of the program Station Control Windows Icons and Toolbars Mouse Control Keyboard Control Tables Editing Adding and Deleting Lines Local menus of tables Configuring Tables Clipboard File Selection Saving Files Record of all operations of station Log Structure of EAS32 Windows Main Station Window Channel Window Chromatogram Window Calibration Window Sample Table Window V

6 Contents EAS32 v First Analysis Chromatogram Measurement and Evaluation Measurement Processing the Analysis Method Chromatogram Fundamental Procedure of Analysis Analysis External Control of Analysis Run Simultaneous Measurement in Several Channels Automatic Functions after Analysis Processing Batch Measurement of Analyses Sample Table Active mode without Control Active Sample Table with Control Sample Table and Its Creation Procedure of Sample Table Measurements Off-line Processing of Analyses - Batch Chromatogram Displaying a Chromatogram Display Characteristics Chromatogram Description and Display Description and Format of Displayed Axes Descriptive Labels and Lines Operations Involving Several Chromatograms - Overlay Resizing and Relocating Chromatograms Mathematical Operations VI

7 User s Guide Contents Three-dimensional View of Chromatograms Chromatogram Modifications Baseline Modifications Peak Modifications Selection of Conditions Restricting Integration Separation Parameters Integration Table Calculations and Calibration Result Table Types of Integration Calculations Non-calibrated Calculations - Uncal External Standard Method - ESTD Description of the Calibration File Main Calibration Table Compound Calibration Tables Calibration Curve How to Create Calibration Creating Additional Concentration Levels Manual Calibration Calibration File Modification Default Parameter Settings Response Factor and Free Calibration Peak Identification in Calibrated Calculations "Reference Peak" Method Resolving Instances of Overlapping Identification Windows Updating Retention Times VII

8 Contents EAS32 v Reports Reporting Procedure Report Style Files Printer Selection Report Style Selection Report Style Modification Export Exporting a Chromatogram and the Corresponding Results as Text Exporting Chromatograms as Vector Pictures Resolving Problems Running the Program Signal Displaying and Measurement Sample Table Processing and Displaying Chromatograms Chromatogram Modification Calibration Calibrated Calculations EAS32 Files and Directories Directories Files Chromatograms (*.PRM) Method Files (*.MET) Calibration Files (*.CAL) Sample Table Files (*.SEQ) Report Style Files (*.STY) Password File (EAS.PSW) VIII

9 User s Guide Contents Configuration File (EAS.CFG) The Desktop File (EAS.DSK) File Sharing File Locking Marking Changes in a File Not Yet Saved Station Settings System Configuration Setting Number of Channels Description of Channels Linking Several Channels User Accounts - Protected Mode Channel Locking User Settings Working with Directories and Projects Channel Directory Selection Projects Backing up and Restoring Files and Projects Backup Restoring Files Glossary List of Pictures Index IX

10

11 User s Guide Introduction 1 Introduction 1.1 Description of the Elemental Analysis Software The Elemental Analysis Software is a universal device for measuring and evaluating chromatograms in both liquid and gas chromatography. The station enables one to process analyses comprising hundreds of peaks whose width may vary from tenths of a second to tens of minutes. Owing to the adjustable range of the inlet A/D converter (from ± 156 mv up to ± 10 V) the station can be directly connected to the output of almost any chromatograph. The station consists of a one- to four-channel internal board or a one- to two-channel external 24-bit integrating A/D converter and a software utility. Thus, in the maximum configuration one can measure simultaneously up to four independent analyses. A brief description of the converters can be found in the Reference Manual. The station collects chromatographic data independently, "in the background", while the user analyses previously acquired data, corrects the baseline, performs calibrated or uncalibrated calculations, or runs another program. Easy collaboration between the station and an autosampler is also possible. Results can be printed on any printer, and the printout layout adapted to the user's needs through a host of adjustable parameters. Tables, data and displayed chromatograms can be exported as files or inserted directly to other programs running under Windows. Calibration files enable the user to carry out calibrated calculations. Each calibration file can contain an arbitrary number of components, each calibrated at up to 20 concentration levels. 1.2 Updating of the program By purchasing the EAS32 you have also obtained the possibility to download all smaller updates free of charge. We are constantly developing the EAS32 according to the requirements of our customers. Should you have any problem with the station, please do not hesitate and let us know. Whether this concerns inadequacies of the existing functions or tips for various improvements, we are pleased to hear the opinion of users, which we take very seriously in our work on software equipment. 11

12 EAS32 v.1.4 Introduction Automatic check of last available version Should your computer be connected to the internet, you may make use of automatic checking of the last available update. In the main window of the station use the command Help Check for Updates. The command compares the number of the last available version on the internet server with the version you have installed. Should a newer version now be available, a dialogue window with the number of the new version appears and you may download the new version. After downloading the Elemental Analysis Software automatically shuts down and runs the installation program, which shall take you step by step through the installation of the updated version. 12

13 User s Guide Station Control 2 Station Control Although working with the station does not differ from working with other programs running under Windows, we present a brief description of some important topics along with some useful advice. 2.1 Windows Main windows The Elemental Analysis Software has a hierarchical structure. When run it displays the main window Elamental Analysis Software with symbols of chromatographs (detectors) connected to individual channels. Each channel consists of up to four main windows (the Channel window proper and the Chromatogram, Calibration, and Sample Table windows). When any of them opens the corresponding icon and name is shown in the bottom taskbar. If you find too many icons displayed in the taskbar upsetting, check off the command Show Windows on the Taskbar in the General card of the User's Options dialog box (open it by the command View - Options from the Channel window). Dialog (Modal) windows Some open windows - called modal or dialog windows - prevent the user from opening any other window until the modal window is closed (an example is the window Open File for selecting files). The fundamental key combinations used to facilitate work with windows: Alt + F4 Close the current window Esc Close a dialog window Alt + Tab Switch between windows of all programs currently running F1 Invoke the context-sensitive help 13

14 EAS32 v.1.4 Station Control 2.2 Icons and Toolbars Icons Icons are small graphical symbols that facilitate invoking some functions or windows. All icons have the corresponding counterparts in menu commands. Toolbars Toolbars are panels containing icons. Toolbars are usually situated below the title bar, but can be moved to any place on the screen. The Elemental Analysis Software contains several predefined toolbars that may be customised (icons can be added or removed); the user can also create his/her own toolbar. Flowchart The channel windows display a special arrangement of icons organised in the same way as typical operations in chromatogram measurement and processing of. The corresponding commands can be invoked in any order - the displayed arrangement merely facilitates orientation and assists one in following the appropriate working procedure. An unconventional element is the triad of 'traffic sign' symbols or corresponding to the first three items of the dialog box Postrun Setting. 2.3 Mouse Control As in most programs running under Windows controlling the Elemental Analysis Software by the mouse is the easiest and in fact almost indispensable method. EAS32 makes use of both the left and right mouse buttons. The fundamental concepts are explained below. Clicking Clicking means pressing the left mouse button at a moment when the mouse cursor (arrow) points to the requested position (icon, button, edit line,...). In most instances it replaces the Enter key. 14

15 User s Guide Station Control Double-clicking Click the left mouse button twice in rapid succession at the same location. It is often used to select a file or to highlight a whole word in the edit line. Clicking with the right mouse button Usually displays a local menu. In a graph one may assign a different meaning to double-clicking the right mouse button by invoking the command Doubleclick from the User's Options - General dialog box. (To open the dialog box invoke the command View - Options from the Channel window.) 2.4 Keyboard Control The station can be operated also from the keyboard. In the following we present the meaning of some keys and key combinations (pressing two keys simultaneously). Keyboard functions of Windows system: Enter Exercises the selected command (the selected command is shown highlighted in the menu; in a dialog box the borderline of the button is emphasised by a continuous or broken line). The required command can be selected from the menu by the cursor keys, in a window by pressing the Tab key. Esc In a dialog box the Esc key substitutes for the Cancel button - closes the box without saving changes. Hides an expanded menu. Tab Gradual selection of commands, parameters, edit lines and buttons in the active window. The selection must be usually confirmed by the Enter key. Spacebar Rapid change of the selected parameter (checking or unchecking). Alt Selection of the first menu item. Alt + letter Rapid selection of a command or parameter that has the selected letter underscored. 15

16 EAS32 v.1.4 Station Control Insert Used to switch between the insert and overwrite regimes. When editing text lines you may use Insert to decide whether new characters will be inserted at the cursor position or will overwrite the characters to the right. Keyboard functions of Elemental Analysis Software: Keys Function Validity F1 F2 CTRL + N CTRL + O CTRL + S CTRL + P CTRL + X CTRL + C CTRL + V CTRL + Z CTRL + SHIFT + Z CTRL + I DEL CTRL + - CTRL + + CTRL + * CTRL + R CTRL + SPACE CTRL + M CTRL + B CTRL + E Help Editing mode of table New file Open file (Open) Save file (Save) Print (Print) Cut (Cut) Copy (Copy) Insert (Paste) Cancel last command (Undo) Repeat last command (Redo) Insert and create lines (Paste Insert) Erase table field Display previous cut-out (Previous Zoom) Display next cut-out (Next Zoom) Display in original size (Unzoom) Start Sample Table (Start) Pause Sample Table (Pause) Resume running of set Sample Table (Resume) Immediate termination of running of Sample Table (Abort) Reset to zero features of unmeasured lines (Reset) Everywhere Tables Chromatogram, Calibration, Data Acquisition Sample Table 16

17 User s Guide Station Control CTRL + T CTRL + 0 CTRL + 1 CTRL + 2 CTRL + 3 Terminate Sample Table after completion of measurement of sample (Stop) Clear any subranges (All Lines) Mark selected row as the starting line of subrange (From Line). Mark selected row as the last line of subrange (To Line). Mark selected rows as subrange (Range Lines). 2.5 Tables Since tables are not standardised in the Windows environment, we present below the essential characteristics of tables used in the EAS32 program Editing Entering new values Values can be entered directly in individual cells, provided the table allows this operation. Note: Some tables cannot be modified, like the table of results labelled Results; Tables that form a part of a file opened for reading only (READ ONLY files), e.g., when a file is used simultaneously in several channels, also cannot be edited. Having first clicked a cell you may enter a new value that replaces the previous one. By double-clicking or pressing the function key F2 the cell is transferred to the edit mode. If a button with an arrow,, is displayed after clicking, the cell represents a list of values prepared in advance - click the arrow and select the appropriate value from the list. Move among the cells by means of the cursor keys. A next row is created automatically once the first cell has been filled in. Correctness of the current row is checked in its entirety during proceeding modifications and the row can be abandoned only after all errors have been corrected. 17

18 EAS32 v.1.4 Station Control If the requested row is not visible, locate it by means of the vertical scroll bar shown to the right or by the cursor keys. You may also enlarge or even maximise the window to see all rows of the table. Changing values in cells without predefined values Click anywhere inside the cell and enter the new value - the old value will be erased. Upon double-clicking anywhere inside the existing value or pressing F2 you can edit the text at that point Adding and Deleting Lines A new line is created automatically after the first cell has been filled in. The user may edit the new line only after the correctness of the preceding line has been checked. The most facile way how to delete a line is to select it by the grey button at the row's beginning and press Delete Local menus of tables Invoke the local menu by clicking any cell with the right mouse button. The local menu contains commands from the Edit menu: The Undo/Redo function used to cancel or reinstate changes effected in the table. Commands for working with the Clipboard: Copy, Cut, Paste, and Paste Insert. Commands for configuring the table - Setup Columns, Restore Default Columns Commands specific for the given table Configuring Tables All tables enable the user to adjust the width of columns, the order of columns, potentially also the sorting method. How to change the column width To change the width of a selected column move the mouse cursor to the right border of the grey field of the heading - the cursor changes its shape and becomes. Press and hold down the left mouse button and drag to change the column width. Double-click at the above location to adjust the column width to accommodate the longest string of text. 18

19 User s Guide Station Control One may also select several columns at once - by changing the width of any of them then changes the widths of all. How to change the order of columns Proceed as follows: Click the grey header of the required column, press again and hold down the left mouse button and shift the cursor to the left or right, depending on where you wish to move the column. As soon as a thin red line appears at the location where you want to move the left border of the column, release the left button. How to sort items in a table Items (lines) of a column are ordered by default according to the first column or the order in which the lines have been created. To reorder a table according to some other column double-click the heading of that column to sort in ascending order; repeat the process to sort the table in the descending order. Some tables (e.g. the Integration Table) cannot be reordered, or reordering must be first permitted by the command Enable Sort from the local menu. Displaying and hiding columns The result tables in the Chromatogram window and the tables in the Calibration window can be extensively reconfigured by the command Setup Columns from the local menu or from the Edit menu. In the subsequently displayed window Setup Columns the user can easily select the columns to be displayed, define their order and, in particular, define own, dedicated columns. Creating of new column with definition of calculation Invoke the command Add Column to open the Add User Column dialog box, where you can create a new column of the table with a predefined calculation formula. 2.6 Clipboard How to use the clipboard? All data in the tables and parameters may be transferred to other places and even to and from other programs using the standard commands Cut, Copy and Paste. It is possible to use the clipboard for copying text headings and lines between chromatograms within the Elemental Analysis Software. 19

20 EAS32 v.1.4 Station Control Note: A typical example is use of the table calculator (Excel) for easy preparation of an extensive Sample Table. You may then easily transfer this using the commands Copy and Paste into the window Sample Table. Copying of data to the clipboard The values of all parameters and the content of any fields of the tables may be transferred to any other places or other programs of the Windows system with the help of the clipboard. The Clipboard is the temporary memory shared within the entire Windows system, into which you enter the selected value using the command Copy or Cut. You then locate these in another place using the command Paste. In some tables you also use the function Paste Insert (keys Ctrl + I ), which does not rewrite the original content of the table but instead adds new lines above the currently selected line. You can find the commands in the menu Edit and in the local menu of the tables. You may also use the standard key combination: command Copy Ctrl + C, command Cut Ctrl + X and command Paste Ctrl + V. Selecting an area to be copied Use the left mouse button and the grey buttons at the top and to the left of all tables to select a contiguous area. All cells passed by the mouse cursor with the left button held down will be highlighted. The grey buttons to the left select the entire row, those on top select the entire column; the top-left button selects the entire table. To select a contiguous area use a combination of the SHIFT key and the cursor key. The combination Ctrl + A or the command Select all from the local menu selects the entire table (see Chapter Local menus of tables, p. 18). Automatic filling in of table fields with repeating values. Upon filling in tables it may occur that you need to fill in an area of the table with periodically repeating information. The Paste function automatically supports such insertion. It is sufficient to copy into the clipboard the information which is to repeat, and then to indicate the area for insertion of data and use the Paste function. Note: Automatic filling in applies to both lines and columns. Should you thus wish to insert periodically repeating lines, remember to insert them whole into the clipboard (including any empty columns). Deleting values At many places selected values can be deleted by the Delete command or the Del key. 20

21 User s Guide Station Control 2.7 File Selection Open any file by the Open command - a dialog box Open X is shown first (where X stands for the file type - a chromatogram, a method, ) A list of all files of the given type from the current directory is displayed on the left (the path is shown in the title bar). Information about the file If you click any file and have the panel with a detailed view of the file open by icon, the items below indicate a detailed description of the file, the name of the author, the version, potentially a preview of the data. Opening a file Use the OK button or double-click the filename to read in or display a file. If the station operates in the password-protected mode and you do not possess the necessary authorisation to access the file, an error message appears. Fig. 1 The Open Chromatogram Dialog Box 21

22 EAS32 v.1.4 Station Control Changing the directory If you wish to open a file from another directory, click the pull-down menu (combo box) Look in to inspect and change the current path. The following icons shown in the top-right part of the window offer the user immediate return to the current directory: - Current project - The common project COMMON (storing the report styles) - Data subdirectory of the current project (storing chromatograms) - Calibration subdirectory of the current project (storing calibration files and standards) Ordering files By default files are ordered alphabetically. To order files by date/time or size when last saved, set a detailed view by icon and sort the files by double-clicking the heading of the corresponding column, or double-click again to sort in reverse order. The ordering is indicated by the icon in the column heading. Filtering displayed files While the filename is being entered in the File Name item only files beginning in the already entered text are displayed. If you wish to find files containing a given text not at the beginning of the filename, start by entering either * in the File Name item or the corresponding number of question marks. Filtering file types Item File Type serves for selecting the type of displayed files. Thus, item All Files (*.*) displays all files in the current directory regardless of whether they have the extension ascribed to the relevant file type. Simultaneous opening of several files If the OVERLAY mode is enabled, several chromatograms can be selected in the Chromatogram window. To effect a contiguous selection click the first file, then press and hold down Shift and click the last file to be selected. All chromatograms will be highlighted and subsequently read in by clicking the OK button or hitting the Enter key. Use the Ctrl key to select other than contiguous files. Press and hold down Ctrl and then click the files to be selected one by one. Confirm the selection by clicking OK or hitting Enter. 22

23 User s Guide Station Control 2.8 Saving Files Use the commands Save or Save As. The former automatically saves changes effected in the file. The latter is used to save the file under a different name and/or in a different directory: the dialog box Save As opens and shows the filename and directory under/from which the file was originally opened. Enter a different name in item File Name, select another directory by means of the drop-down item (the so-called combo box) Look In and use the item File Type to save the file in another format. Note: The File Type item converts files between the EAS17 and EAS32 in both directions. Fig. 2 The Save As Dialog Box Note: It is not suitable to save files outside of the project folder. 2.9 Record of all operations of station Log The Elemental Analysis Software keeps a log of all operations the station performs. Records of the station s operations are saved in the sub-folder LOG of the main folder of the station. All operations performed during one day are saved into the table Daily 23

24 EAS32 v.1.4 Station Control Log, indicated with the date. In addition to this you may display Session Log with the operations performed since the last running of the station. A table of all modifications of the chromatogram the Chromatogram Log, which is a component of the chromatogram occupies a special place here. What is the log useful for? Logging of activities of the station is one of the essential steps for fulfilment of the conditions of Good Laboratory Practice (GLP). Thanks to the information recorded in the log table we may easily analyse the conditions leading to errors or unstable states of the station. For this reason the program also contains implementation of a function which enables you to send information on the setting of the system and a record of the station s operations in the last two days in the case of incorrect termination of the running of the station. Note: Before sending messages with information you have the possibility to check and modify their content (e.g. add a commentary for user support). The message is sent only after you approve its sending. The log also serves as a diary. For example, one week ago you performed an analysis and exported the results into a file. Now you can t remember the name of the file. No problem. Search in Session of Logs for the given period, arrange the lines according to type of events and check the exports. 24

25 User s Guide Structure of EAS32 Windows 3 Structure of EAS32 Windows The Elemental Analysis Software is organised in a hierarchical structure. When the station is run the main window appears first and displays symbols of up to four chromatographs (detectors). Clicking the chromatograph icon and, if need be, entering the user's name or password opens the Channel window earmarked for measurement and processing of analyses with the connected detector. Note: The concept of the EAS32 is channel/detector oriented; it means that individual channels are fully independent and each has only one detector assigned at any given time. For multidetector measurement, e.g., on a gas chromatograph, the interrelated channels can be synchronised to perform analyses measured with several detectors. 3.1 Main Station Window The fundamental purpose of the Main Window is selection of the channel to be used in a particular measurement, but the window can be also used to configure the station, select the base directories for data storage, set up the digital outputs and select the method of reporting. Most commands in the Main Window are enabled only when all Channel windows are closed. 3.2 Channel Window All tools necessary for performing analysis are brought together in the Channel window. The EAS32 can simultaneously operate with up to four channel windows, in other words, record up to four chromatograms at the same time. Each Channel window contains a Status Table and a graphical scheme of analysis processing. Clicking individual icons in the scheme or invoking commands from the menu opens windows earmarked for performing the appropriate actions with the selected chromatograph (detector). Each Channel window has its own independent windows; accordingly, measurements in progress in different channels can be monitored simultaneously. Channel windows differ in the colour of the connecting lines in the scheme of data processing, the detector name shown in the title bar, potentially in the project (= directory) name in the status bar. 25

26 EAS32 v.1.4 Structure of EAS32 Windows 3.3 Chromatogram Window This is the fundamental window for displaying, modifying and evaluating chromatograms. Open the window by clicking icon in the Channel window or at the end of menus in other windows. Opening a chromatogram To open a chromatogram, click the icon and in the opened window double-click the corresponding filename. Open the Result Table of the displayed chromatogram by the command Results - Result Table. Refer to chapters 6 - Chromatogram on page 53 and 7 - Calculations and Calibration on page 74 for description of chromatogram modifications and evaluation. 3.4 Calibration Window The window enables calibration curves to the created, modified and displayed. Open the window by clicking icon in the Channel window or at the end of the menus in any other window. Refer to the chapter 4 - First Analysis on page 27, section Calibration, for hints how to create and use a simple calibration. 3.5 Sample Table Window Open the window by clicking icon menus in other windows. in the Channel window or at the end of the 26

27 User s Guide First Analysis 4 First Analysis To explain the fundamental practical elements of work with the Elemental Analysis Software we present here a description of a model analysis. The description by no means covers all possibilities of the station setup. We assume the Elemental Analysis Software has been already installed and connected to a chromatograph (autosampler). Running the program Its ability to start measurement immediately without protracted preparations and setup ranks among the fundamental advantages of the EAS32. Run the program by clicking icon on the desktop or selecting the item Programs Elemental Analysis Software in the MS Windows Start menu. Click icon of either the gas or liquid chromatograph. Signal monitoring Click icon to display the detector signal and check the "drift" and noise. Then close the window either by the button or by invoking the command File - Exit. Measurement and processing parameters All parameters that decide on and describe chromatogram measurement and evaluation conditions are stored in a method file. Some parameters (e.g. voltage range, sampling rate, etc.) must be set prior to the start of analysis, and the station prevents their change during the measurement as it might corrupt the results. If you point the mouse to icon it changes to. Clicking any of the four icons displays the contents of the current method in the Method Setup window. The method file is divided into the following sections: Measurement - description of measuring conditions: duration of analysis, the column employed, etc. Acquisition Integration Calculation - Parameters governing signal measurement: voltage range, sampling rate,... - integration parameters, integration table of modifications, etc. - the calculation parameters used to set up the calibration calculations To select a different method file, invoke the command File - Open or click icon. Start of analysis Invoke the Sample Table dialog by means of Fill in the lines you want to measure. icon from the Channel window. 27

28 EAS32 v.1.4 First Analysis Fig. 4 Sample Table window Run the analysis by invoking the icon. The inscription RUNNING will be shown in the line beneath and the actual time of analysis above icon. Monitoring the Analysis To monitor the analysis, click icon. Fig. 5 The Data Acquisition Window Press and hold down the left mouse button, drag and finally release the mouse button to select the area to be zoomed in. Use the Unzoom icon or double-click anywhere inside the graph to restore the original size. The analysis in progress can be compared with an already completed chromatogram, e.g., with the chromatogram of a solvent, a calibration standard, etc. With the command File Set Background Chromatogram you can select which chromatogram is to be displayed on the background of the window Data Acquisition. Note: The background chromatogram on the background is shown in grey colour in order to enable replacement with the measured signal. 28

29 User s Guide First Analysis Note: The chromatogram on the background is displayed after running of measurement. Preparation for analysis termination Prior to analysis termination is it advisable to check the topical integration parameters, either by clicking icon or by the command Method - Integration. Fig. 6 The Method Setup - Integration Window Note: Peak Width and Threshold are the most important parameters. In simple terms one can state the following: Peak Width specifies that all peaks significantly narrower than the value of this parameter will be ignored, while Threshold decides on the noise level, in other words, only peak at least twice as high as the value of Threshold will be detected. In reality the situation is somewhat more complicated since interplay of the two parameters and the peak shape is also involved. Note: The Peak Width parameter in fact decides on the compression (the number of integrated data samples) of the resulting chromatogram and, accordingly, on the extent of signal smoothing and the processing rate. Higher value means higher compression and vice versa. 29

30 EAS32 v.1.4 First Analysis Analysis termination Analysis can be either terminated by the Stop command, or terminates automatically once the time specified by the parameter Enable Autostop in the Method Setup - Measurement window expires. At that moment the measured data are integrated, evaluated, and saved. How to set variables into the name of the file? Experienced users often remember the names and meaning of the variables used and enter them manually. For entering less common variables or for easier entering you have the assistant for selection of variables available in the window Sample table. 1. Place cursor in the File Name column in the place where you wish to insert the variable. 2. Click on the icon next to the line for entering the file name. 3. A list of variables shall appear, including a brief description of the meaning.. 4. Place in the file name by clicking on the required function. Displaying and evaluating the chromatogram The completed chromatogram will be displayed automatically in the Chromatogram window. The user can open the window by clicking icon and display the chromatogram by icon. To prevent the chromatogram from being displayed automatically change the symbol at icon to. As in the window for monitoring the signal, you can enlarge any part of the chromatogram here by clicking on the left pad of the mouse, dragging and releasing the pad at the moment of reaching a suitable cut-out. Manual peak modification or repeated processing (reprocess)? In the event the baseline is unsatisfactory, the user can "fine-tune" the chromatogram manually or reintegrate the whole chromatogram anytime. Manual modifications are accessible through two toolbars shown to the left or by the corresponding command from the respective submenus Chromatogram - Baseline or Peak. Modifications effected are stored in the table of integration results and can be changed or deleted anytime. Reintegration proceed automatically when the user changes a value in the Parameters section from the right-hand part of the Integration card. Access to the card is through the command Method - Integration. 30

31 User s Guide First Analysis Topical character and consistency of data The EAS32 automatically maintains all data topical and consistent; accordingly, a baseline modification is immediately reflected in the Result Table. A change of any parameter updates all ensuing calculations and the displayed results. For this reason the station does not offer commands like Update, Recalculate and similar. Print To facilitate printing the results all major windows contain icons,, and. Click icon in the Chromatogram window. Select and set up a printer by the Printer command situated in the right-hand part of the opened window, and select a suitable report style by the command Open. The file containing the report style is in fact a prescription specifying which information will be printed and how. Information is divided into segments contained in the corresponding tabs. Each tab enables one to select detailed layout of the segment. To include a segment in its entirety in the printout, check item Print situated in the top-left corner of the tab, or activate the check mark before the tab name by double-clicking the tab label with the left mouse button. Icon serves for displaying print preview, icon starts printing. Note: At each "print" command across the application different report styles are preset (and can be changed) in a manner ensuring that the contents of the relevant window are printed as far as possible. Calibration Creation and use of calibration files occupies a substantial part of the EAS32. Calibration curves created by means of calibration standards are stored in separate calibration files; each calibration file can contain a practically unlimited number of calibration curves prepared at up to twenty concentration levels. Prior to using calibration results the user must: 1. Create calibration curves and save them in a calibration file. 2. Assign a calibration file to the chromatogram. Creating a calibration curve Click icon or invoke the command Window - Calibration. 31

32 EAS32 v.1.4 First Analysis In the opened Calibration window create a new calibration file by clicking icon or invoking the command File - New. Click icon or invoke the command File - Open Standard to read in a chromatogram representing a calibration standard. Click icon to transfer the available data of all peaks to the calibration table. In the displayed table the names of compounds in the first column can be changed. The known weights of individual compounds can be then entered in the Weight column beneath the calibration level. Provided you followed the above directives, clicking the tab with the name of the required compound will display a straight line representing a single-point calibration dependence. Finally, save the created calibration curves in the file under the selected name by invoking the command File - Save. Note: The station enables multilevel calibration curves to be established and effected by a selectable algorithm, all either automatically or by manual selection of individual compounds, including interactive filling in and checking of all entered data items. The shape of the calibration curve, the type of response, the size of the identification windows and improved identification by means of reference peaks can be all selected for any compound. Using a calibration curve The simplest way how to check correctness of calibration is to apply it to the original, underlying chromatogram. To do it, pass to the Chromatogram window (e.g., by clicking icon ) and, once there, open the window for chromatogram selection by clicking icon. Since calibration standards are stored in subdirectory Calib, first select that subdirectory (e.g. by clicking icon in the top-right corner of the Open Chromatogram window) and select the chromatogram from which the calibration curve was created in the first place. The next step is to decide on the method of calculation: display the Results tab and select your calibration file by the command Set. The Result Table should now contain the known weights of the entered compounds including their names and the corresponding percentages. Zero weight will be stated for peaks for which no calibration standard was identified according to the retention time. Identified peaks for which no calibration is available will be labelled in the Peak Type column by inscription Free. Note: It follows from the above that calibration curves are stored neither with the chromatogram nor with the method - each chromatogram only contains a reference to the corresponding calibration file. This approach is not standard with 32

33 User s Guide First Analysis other applications. The advantage consists in facile calibration and changes in those instances where a single calibration file is used for several chromatograms. The necessity to save the calibration file under a different name in case of changes that must not affect chromatograms already evaluated (archived the original results) is a certain drawback, though. 33

34 EAS32 v.1.4 Chromatogram Measurement and Evaluation 5 Chromatogram Measurement and Evaluation This Chapter offers comprehensive review of all possibilities how to measure chromatograms or series of chromatograms. 5.1 Measurement In this connection measurement comprises conversion of the analog detector signal to digital form by means of an A/D converter, digital integration of the measured data, and storage of the results on the hard disk. Each Channel window can exist in any of the following three states, regardless of the other Channel windows: STOP - No measurement is in progress and starting it by an external signal is prohibited. WAITING - No measurement is in progress and the station awaits the external "start" signal. RUNNING - An analysis is in run. Measurement with the EAS32 comprises the following steps: The A/D converter transforms the analog signal to a 24-bit binary number representing the integral value over a given time. Note: A true integral is indeed involved owing to the design of the converter, which permanently integrates the input voltage, in contrast to a sampling converter with approximation transfer, which samples the voltage at discrete time intervals. The above binary number then passes through a digital filter that filters out all parasitic components originating, e.g., in power voltage fluctuations. Individual binary numbers (data items) are read from the converter at a rate ranging from 10 to 100 data items a second; each partial integral occupies four bytes. Note: With some chromatographs a signal already digitised by an internal A/D converter can be sent to the EAS32, usually along the RS-232 serial line. Thus, if the corresponding driver is available, order the EAS32 without the A/D converter. Data samples are stored temporarily in operating memory and sent to the hard disk about once a minute; in either the data or the calibration subdirectory of the current 34

35 User s Guide Chromatogram Measurement and Evaluation project a file $CHXRUN.RAW (where X stands for the channel number) is thus created for storing the so-called raw data. This ensures that very long chromatograms can be measured since the capacity of the hard disk is much higher than the capacity of the operating memory. Selecting the sampling rate In the area of extreme measurements on capillary columns and microcolumns in particular you would be glad to make use of one of the four preset sampling rates: 10, 25, 50 or 100 samples per second. The setup is a part of the method file and can be changed from the Channel window by the command Method - Acquisition - Sample Rate. Changing the sampling rate affects the chromatogram size. Note: The available scopes of sampling speed depend upon the type of converter used. 5.2 Processing the Analysis Processing chromatographic data means identification of all peaks in the chromatogram and drawing the proper baseline. To process chromatographic data the EAS32 is equipped with a standard algorithm that requires the knowledge of the parameters Peak Width and Threshold. They are a part of the method file and upon analysis termination are transferred to the chromatogram. Peak Width The parameter decides on the peak width at half height (expressed in minutes) of peaks occurring in the processed analysis. The value should approximately correspond to the parameter W05 (peak width at half height). For peaks of different width the value should be set according to the narrowest one. Threshold The parameter discriminates between useful signal and noise. It is entered in millivolts and is independent of peak width. The height of peaks still to be included in integration, measured between the peak beginning and apex, should be at least twice as high as the value of this parameter. Both parameters fundamentally influence the quality of the resulting chromatogram with regard of peak detection (determination of peak beginning and end and the correct baseline). Accordingly, in creating a methodology used for each class of analyses pay appropriate attention to the optimum setting of the two parameters. Although the setting is not too critical (the result will be the same within a certain range of values), their incorrect selection must be subsequently corrected by operations with the chromatogram 35

36 EAS32 v.1.4 Chromatogram Measurement and Evaluation (see Chapter Chromatogram Modifications on page 62). These additional operations should not substantially change the selected baseline, rather eliminate some phenomena that the peak detection algorithm is unable to cope with (e.g., tangential separation, identification of the solvent peak, ban on detection during the period of column switching, etc.). Some practical guidelines referring to setting up these parameters are in Chapter Processing and Displaying Chromatograms on page 101. In general, the width of detected peaks (expressed as the distance between the peak onset and end) is directly proportional to Peak Width and inversely proportional to Threshold. Moreover, the Threshold parameter prevents erroneous assignment of noise to spurious peaks. In addition to the above two parameters one can determine the start of integration. Integration Start The value of this parameter determines the time of analysis when the integration algorithm will be initiated; in other words, no integration will take place prior to that time. This is the main difference compared with the command Baseline - Lock that merely cancels already integrated peaks within the specified chromatogram segments and never reintegrates the chromatogram in its entirety Method The method file constitutes a tool used to achieve facile description and setting of all measurement conditions and to attain high reproducibility of measurements performed with the EAS32. The method is divided into a number of sections, of which each is reserved one tab of the Method Setup window. Measurement - Section with description of measuring conditions and possible setting of length of measurement or automatic deduction of solvent. Acquisition - Section of parameters related to signal measurement - attenuation, sampling rate, control of digital outputs through the Event Table, etc. Integration - Section of integration parameters, a table of integration modifications, etc. Calculation - Section summarising calculation parameters for setting the type of calibration calculations. The method file is available from the Channel window by commands contained in the File menu. 36

37 User s Guide Chromatogram Measurement and Evaluation Note: Carefully select the method for measuring a chromatogram, in particular with regard to attenuation, duration of analysis, the limiting integration parameters, the contents of the integration table and the selection of the calibration file. It is advisable to check in particular the parameters from the Acquisition tab prior to starting an analysis, since they cannot be thereafter modified! Upon analysis termination a copy of the method file is stored with the created chromatogram. Thus, most parameters are available and can be changed for the already measured chromatogram in the Chromatogram window through the commands from the Method menu and the Results menu. Note: The layout of individual commands does not correspond here to the tabs of the Channel window, mostly because the parameters from the Acquisition tab serve merely for information and cannot be changed Chromatogram Raw data processing results in the creation of a chromatogram (file extension PRM). Chromatograms consist of three basic parts: a copy of the method, the raw data, and the chromatogram proper with identified peaks and the baseline marked. Chromatograms can be displayed in either the Chromatogram or the Calibration window, printed, or the file contents exported to the Clipboard or to a text file. Chromatograms of individual projects are stored in either the data or the calibration subdirectory. Click icons and in the Open Chromatogram window to switch rapidly between the subdirectories. Note: If you prefer not to have chromatograms separated in the above manner, select identical names of the two subdirectories when establishing a new project in the Project Setup window. To obtain detailed information about the current chromatogram invoke the command Method - Measurement from the Chromatogram window. 5.3 Fundamental Procedure of Analysis The following chapters present directives informing about various methods of analysis processing. The basic procedure was already summarised in Chapter 4 - First Analysis on page 27, and copious information is available from Chapter Signal Displaying and Measurement on page

38 EAS32 v.1.4 Chromatogram Measurement and Evaluation Analysis This chapter describes a comprehensive procedure of analysis processing, taking into account most options the station offers. Some additional variants are discussed in subsequent chapters. Channel selection In the Main window click the channel icon representing the detector you wish to use in the analysis. Fill in your name or, in the protected mode, enter the password. The OK button then opens the corresponding Channel window. Setting up measurement conditions All parameters and data items referring to the measurement are summarised in the Measurement and Acquisition tabs of the current method; accordingly, display them first by clicking either of the icons and i, or by the command Method - Measurement (Acquisition). The name of the current file is shown in the title bar and in the Status Table. Setting up the maximum input voltage The input range (attenuation) is defined by the Range item in the Acquisition tab, and must be higher than the maximum output voltage of the employed detector. On the other hand, it is certainly inappropriate to measure small voltages with, say, the 10-V range, since it would reduce the measurement resolution and accuracy. If you intend to measure negative voltages, check the Bipolar item. Monitoring the baseline drift and noise Open the Data Acquisition window by icon or the command Monitor - Detector Signal. The detector signal will be identified automatically and displayed within the values given by items Time and Voltage. In the event the displayed range is unsatisfactory, change the values in items Time and/or Voltage and confirm by pressing Enter - the signal will be displayed with the selected magnification. Use the left mouse button to zoom in on any cut. To resume at the original magnification specified in Time and Voltage, double-click with the left mouse button. 38

39 User s Guide Chromatogram Measurement and Evaluation The station stores the selected cuts; to see them use the commands Previous and Next Zoom, respectively. Note: In a zoomed-in cut the window is no longer redrawn automatically (as is the case when the base range of the window is exceeded). Monitoring the analysis Reopen the Data Acquisition window by icon or the command Monitor - Detector Signal. By default the detector signal is not identified during analysis, but the fixed range as defined by items Time to and Voltage to is always displayed. When the preset values are exceeded the window is not redrawn automatically. You can also compare the analysis in progress with a completed chromatogram, e.g., that of the solvent, a calibration chromatogram, etc. Invoke the command File - Open Background to select the chromatogram to be displayed in the background of the Data Acquisition window in grey, but only after the current analysis is in run. Note: The values entered in items Time and Voltage do not limit in any way the scope of the acquired data. The maximum voltage range is defined by items Range and Bipolar on the Acquisition of the Method Setup window, and the duration of analysis may be restricted only by the parameter Enable Autostop from the Measurement tab of that window. Setting up the processing parameters The measured chromatogram will be integrated and evaluated according to the parameters specified in Integration tab of the current method. Do not forget to check the following parameters in particular: Peak Width - Enter the width of the narrowest peak expected (in the Integration tab) Threshold - Enter the half height of the lowest expected peak (in the Integration tab) Calibration File - Identifies the calibration file containing the table of compounds used for their identification as well as the calibration curves used to calculate the actual weights. Preparation for analysis termination Check the Setting - Postrun command from the Channel window as to activities to proceed after the measured data are processed (see Chapter Automatic Functions after Analysis Processing on page 44). 39

40 EAS32 v.1.4 Chromatogram Measurement and Evaluation The automated numbering option is very useful in normal operation. The station uses a set of variables preceded with the character %. Upon analysis termination each such variable is replaced by the topical value. Variables may be combined at will - the topical value is shown in parentheses above the edit cell and in the status table of the Channel window. For additional details see the Reference Manual, Chapter Chromatogram File Name. Assistant for setting file name By clicking on the icon next to the line for the File Name column in the Sample Table window you can open the assistant for creation of names of files from individual variables. Simply select the required function, click on it and the assistant automatically adds the corresponding variable to the file name. Note: In some instances the application saves the chromatogram under a special name, using the first free eight-digit number starting at *, specifically in situations where there is a risk that measured data might be lost or existing data overwritten, to with: - the user has cancelled the invitation to enter new chromatogram name by the Cancel command, or - the application was unable to display a message requesting that the new chromatogram name be entered, since some other message was displayed at that time. End of analysis Activate the Stop button. The analysis will then terminate and the measured data saved, potentially evaluated, displayed and printed, depending on the setting effected in the preceding two items. Activation of the Run button terminates the analysis in the same way and runs the next analysis - continuous measurement of a series of analyses. Note: You may also terminate analysis with the commands Analysis - Stop, Abort (icons, ) in the window Data Acquisition. Displayed results Displaying the chromatogram automatically in the Chromatogram and Calibration windows depends on the setting of the corresponding items in the dialog box Postrun Setting. Open the windows manually by means of icons and and display the chromatogram by invoking the commands File - Open and File - Open Standard, respectively. 40

41 User s Guide Chromatogram Measurement and Evaluation Data evaluation Invoke the Results - Result Table command to display the table of integration results in the Chromatogram window. See Chapter 7 - Calculations and Calibration on page 74 for a detailed description including the procedure used to obtain calibrated results. Presentation of results The results of an analysis can be automatically printed, exported (see Chapter 8 - Reports on page 90) and also processed by an automatically run program, all by appropriate setting of items on the dialog box Postrun Setting - see Chapter Automatic Functions after Analysis Processing on page External Control of Analysis Run Analyses can be also run and terminated by an external signal - a chromatograph button, an auxiliary button or a sampling valve (depending on your configuration and program installation). The item External Start/Stop in Acquisition tab of the dialog box Method Setup enables external control. Items in the synonymous section decide on the external control mode. Fig. 7 Section External Start/Stop of the Acquisition tab Items Down and Up decide whether the station will react to the ascending (leading) or descending (trailing) edge of the external pulse. The item Start Only specifies that the external signal will only start analyses, to be subsequently terminated manually, either by the Stop command or by checking the Enable Autostop parameter. The Start-Restart item decides on whether the external signal will stop the currently running analysis and immediately start the next one (continuous series of analyses). The Start-Stop item specifies that the external signal will only terminate the currently running analysis, and the start of the next analysis will wait for an additional external signal. 41

42 EAS32 v.1.4 Chromatogram Measurement and Evaluation Simultaneous Measurement in Several Channels The station allows for synchronised, simultaneous measurement in several channels, thus facilitating work with several detectors. Coupled control of several channels is set in the dialog box System Configuration by the Link with command in the tab of the corresponding Channel x by checking the interconnection with the required channels. The following commands are then coupled: Run, Stop, Abort, and Autostop. Moreover, an external signal received at any of the coupled channels starts or stops the remaining channels as well. The Snapshot command processes only data from the current channel. Should the Enable Autostop items be set differently in individual channels, the station will use the shortest of all preset times. In the event a batch measurement (Sample Table) is run on any of the channels, the remaining coupled channels are run as well. To secure facile evaluation of analyses measured in different channels in a single Chromatogram window, it is recommended to identify the same project in all channels by the command File - Project - Open. Event Table creation and modification To display the Event Table invoke the Method - Acquisition command in the Channel window. Fig. 8 Events Table A new line is appended automatically once the previous line is filled in correctly. Manual setting of control outputs Control outputs can be also managed manually by the command System Digital Outputs in the Main window. 42

43 User s Guide Chromatogram Measurement and Evaluation Fig. 9 The Digital Output Control Dialog Box for the INT7 board Invoking the command opens the dialog box Digital Output Control XXX Board (where XXX stands for the name of the A/D board used or selected). The first column, Initial, is earmarked for setting up the default state of the outputs each time the program is run. The second column, Current, sets the topical state of the outputs and enables the user to change it immediately. A Description column is provided for entering notes that explain the meaning of individual digital outputs. Thus, the preset description highlights the use of some digital outputs in Active Sample Tables Automatic Functions after Analysis Processing The station enables subsequent activities to be performed each time an analysis is terminated or for each chromatogram in batch processing by the Batch command. Setting is through the command Setting - Postrun from the Channel window. The command opens the Postrun Setting dialog box containing commands for automatic display of the chromatogram, report printing and exporting the chromatogram, and for running another application. The setting of the first three items corresponds to, and can be controlled by, the "traffic signs" and at the corresponding icons, and in the Channel window. 43

44 EAS32 v.1.4 Chromatogram Measurement and Evaluation Fig. 10 The Postrun Setting Dialog Box Automatic displaying of the chromatogram Set automatic displaying of the chromatogram in the Chromatogram window by means of either the item Open Chromatogram Window or the traffic sign at icon in the Chromatogram window. In the event the OVERLAY mode is operative, chromatograms will be displayed one by one and overlaid. Use the Open Calibration Window item or the traffic sign at icon to set automatic displaying of the chromatogram in the Calibration window. Automatic displaying is suspended if the OVERLAY mode is not operative and a chromatogram is displayed containing some as yet not saved changes, or if the number of displayed chromatograms has reached 100. Automated reporting Use the Print Results item or the traffic sign at icon to enable automatic printing of the chromatogram according to the report style set by the File - Report Setup from the Channel window. Automatic export Check the Automatic Export item to ensure that selected data will be exported automatically to a file or to the Clipboard of the Windows operating system. The export mode depends on the setting of items in the dialog box Export Chromatogram - open it by either of the commands Setting - Export from the Channel window or File - Export from the Chromatogram window. Automatic running of another application If the line Run Program contains a name of an application (say EXCEL.EXE) and the line Parameters contains optional parameters of that application (for Excel e.g. the name of the working file or macro file), that application will be run automatically. Use 44

45 User s Guide Chromatogram Measurement and Evaluation the button to search for the required application including the relevant full path. If the item Only with export is checked, the application will be run only when item Automatic Export is also checked Batch Measurement of Analyses Sample Table The station enables batch measurement of analyses to be performed, in particular in connection with autosamplers Active mode without Control Active mode is used to increase reliability of co-operation between the station, the autosampler and the chromatograph when an autosampler without autonomous control is employed. The autosampler must be equipped with one starting input and one confirmation output. Once a Sample Table has been run by the Run command from the Sample Table window, the station sends an authorising signal and awaits its confirmation by the autosampler; after its receipt the station starts measurement. The run terminates after expiration of the time specified by parameter Enable Autostop from the corresponding method and the station remains in the IDLE state for a period defined by the Idle Time parameter. Thereafter a new authorisation signal is sent to the autosampler and the station awaits its confirmation. The above cycle is repeated for all measurements listed in the Sample Table. If the Sample Table has to be updated, a Sample Table may be suspended by the Break command; the command can be invoked anytime, but the Sample Table will be only suspended the next time the station goes over to either the IDLE or the WAITING state. Invoke the Resume command to continue the Sample Table. Conditions underlying start of an Active mode 1. Checked item Active Sample Table in the Sample Table - Options dialog box. 2. Enable Autostop enabled in each method included in the Sample Table. 3. A logical loop created: the relevant digital output connected to the confirmation autosampler input, the output signal from the autosampler connected to the corresponding external digital output. 45

46 EAS32 v.1.4 Chromatogram Measurement and Evaluation The assignment of digital outputs is following: Channel Output Output pin Nr. on the connector INT7 INT7 Relay 1 OUT , 30 2 OUT , 33 3 OUT OUT All DGND 19 Note: Should you use an Active Sample Table, make sure you do not process these outputs in the Events Table. One injection cycle Measurement (analysis) (STOP) WAITING IDLE WAITING RUNNING (BREAK) IDLE WAITING RUNNING Start program Run Sequence 1st inj. enable 1st injection Autostop Resume 2nd inj. enable 2nd injection time OUT0-6 (digital output) IN1-4 (digital input) Autosampler broadcasts injection Given by the value Initial from thedigital Outputs window The program returns the original digital output Fig. 11 Time Scheme and Succession of Signals in an Active Sample Table Active Sample Table with Control We supply a special program module for some selected types of autosamplers; the module provides for direct selection of the injection Sample Table and injection time. The interconnection for Active Sample Tables is then usually supplemented by a serial communication cable between the PC and the autosampler. A detailed description is in a separate manual Sample Table and Its Creation Sample Table measurement is defined in a Sample Table, where each line specifies the measurement of one or several samples. The table including the auxiliary data is stored in a Sample Table file. Thus, to perform a certain measurement it suffices to select a suitable Sample Table file and start the Sample Table. 46

47 User s Guide Chromatogram Measurement and Evaluation Displaying the Sample Table window To display the Sample Table window click icon or invoke the Analysis - Sample Table command from the channel window. The last used Sample Table file will be shown. Fig. 12 The Sample Table Dialog Box Editing a Sample Table A new line is appended automatically each time item Sample. At the same time the following remaining mandatory items are filled in automatically: Weight [mg] - must be entered for standards. File Name - the name under which the chromatogram will be saved. You can compose it from variables. Std, Lvl - the calibration standard and the level to be calibrated. Standard Name - the name of the standard from the standard table to be used in measuring the analyses performed according to the given line. Save the completed Sample Table file by the command File - Save or Save As. Newly created sample table must be saved by Save ( command prior to measuring the analysis. ) or Save As Until you save it the Start ( ) command will be blocked. Selection of initial and final lines of measurement In practice it may occur that you have a defined Sample Table for more extensive analyses but you currently only need to use a few lines of this table. One possibility is to indicate these lines, copy them into the clipboard using the command Copy (Ctrl + C), create a new Sample Table using the command File New in 47

48 EAS32 v.1.4 Chromatogram Measurement and Evaluation the window Sample Table and insert the copied lines into the new table using the command Paste (Ctrl + V). In this manner it is also possible to copy lines which are not adjacent by holding down the Ctrl key whilst indicating lines. Should you however wish to use a connected section of the Sample Table only once or are using another section, it is more suitable to set the section of the table to be used (Selection) in the Sample Table Options window, thus on which line the Sample Table is to commence From Line and on which it is to terminate To Line. The selection of sub range is available also directly from the sample table window by selecting the line(s) and select a command from the context menu (right mouse click) or by means of keyboard shortcut CTRL + 1 From Line, CTRL + 2 To Line, CTRL + 3 Range Lines Procedure of Sample Table Measurements 1. Opening the Sample Table window To open the Sample Table window either click icon Analysis - Sample Table. or invoke the command 2. Selecting a Sample Table file Open the requested Sample Table file by the command File Open or by clicking icon. 3. Checking the Sample Table and the files used Check all parameters in the displayed Sample Table, or check all method files used: click anywhere in a line with the required method and display its contents either by the command Sample Table - Edit Current Method or by clicking icon. 4. Running the Sample Table To run the Sample Table use either the command Sample Table - Run, icon. The Sample Table assumes the WAITING state; only then is it possible to start the measurement of the first sample by an external autosampler signal. 5. The measurement process Measurement proceeds by individual lines of the table. Each line may define several measurements of the same sample in item I/V, or measurements of more than one sample under identical conditions - items SV and EV. 48

49 User s Guide Chromatogram Measurement and Evaluation 6. Information about the running Sample Table Information is displayed both in the status bar at the bottom of the Sample Table window (display it by the command View - Status Bar) and in the Status table of the Channel window, always as the name of the analysis currently in run, the position number, potentially the serial number of the injection from that position. The Channel window also shows the method used and the first two items of the chromatogram header, Sample and Sample ID. 7. Measurement processing in Sample Table analyses The method file listed in the current line of the Sample Table in column Method Name decides on the method of measurement and processing of the sample. The style of the report, if requested, is given by item Report Style. If no style is specified, the station will use the report style set in the Channel window at icon. Specifics concerning chromatogram displaying, printing and exporting, or running other applications also during the Sample Table are defined in the settings effected in the Postrun Setting or by the traffic signs and in the Channel window, and are thus common to the entire Sample Table Off-line Processing of Analyses - Batch The station provides for batch processing of the chromatograms already measured. Batch processing requires selection of chromatograms that will be subject to the operations analogous to those included in the Postrun Setting window. The command Analysis - Batch from the Channel window opens the Batch dialog box. Fig. 13 Batch Window 49

50 EAS32 v.1.4 Chromatogram Measurement and Evaluation The File Type command selects processing of chromatograms, calibration standards or Sample Tables. A list of the corresponding files in the current project is then shown in the left column. Sample Table processing If a Sample Table is to be batch-processed, all chromatograms listed in the Sample Table in column File Name will be included. Missing chromatograms (not measured or deleted) will be ignored. Note: In the event the File Name column contains variables that prevent the relevant chromatogram to be unequivocally identified (date, time, serial number,...), chromatograms are sought according to a hidden list of actually measured chromatograms. Also here processing comprises only the selected operations listed in the right-hand part of the Batch window. Comprehensive Sample Table processing The Complete Processing command performs repeated processing according to the topical contents of the Sample Table. The headers of all chromatograms will be overwritten and the method actually specified in the Sample Table will be employed; calibration will be carried out if required. Processing according to the current method If you wish to process chromatograms according to the current channel method, check item Reprocess by Channel Method - all parameters listed in the Integration tab will be used. Selection of several chromatograms to be processed To select several files hold down the Ctrl key while clicking with the left mouse button (an additional click cancels the selection). To select a contiguous sequence of files click the first file to be selected with the left mouse button, hold down Shift and click the last file to be selected - all interjacent files will be selected. Notice the self-explanatory commands Select All (Ctrl + A) and Unselect All. Processing order The items in the list of files can be sorted alphabetically by filename (Sort by: Name) or by the saving date (Sort by: Time), either in ascending (item Normal) or descending order (item Backward). 50

51 User s Guide Chromatogram Measurement and Evaluation Processing proper Start the processing by the command Proceed. If no chromatogram or Sample Table is selected, the command will merely save the current settings of the Batch window. Items Open Chromatogram Window through Run Program are the same as those in the Postrun Setting window- see Chapter Automatic Functions after Analysis Processing on page 44 for additional details. 51

52 EAS32 v.1.4 Chromatogram 6 Chromatogram The EAS32 enables the user to apply a number of operations to the chromatogram: baseline modification including the method used for drawing the baseline, shifting peak beginnings and ends, creating new peaks, and selection of limiting parameters for peaks to be included in integration. The effected operations are stored with the chromatogram and may be used in subsequent analyses, thus enhancing reproducibility of the obtained results. Owing to the intuitive mouse operations and the auxiliary icons provided, all operations are simple and facile. Any part of the chromatogram can be zoomed in to define the exact location of the intended operation. Several chromatograms can be displayed simultaneously by the commands from the Overlay submenu and subsequently subjected to mathematical operations. 6.1 Displaying a Chromatogram Chromatograms are displayed in the Chromatogram window, opened by icon from the Channel window, or by the command Window - Chromatogram from other windows. It is also possible to open the window automatically after the end of measurement or in batch processing by the Batch command. Unless the OVERLAY mode is enabled, the currently displayed chromatogram will be closed prior to displaying a new one. The size of the chromatogram is such that if fills in the entire graph area. To close the displayed chromatogram(s), invoke either of the commands File - Close or Close All. Indicative graph In addition to the main graph a smaller, indicative graph can be displayed by the command Preview Graph from the Display menu or by means of the local menu. The chromatogram is always shown in the indicative graph at the base magnification and contains the cut, if any, shown in the main graph. Zooming in Point with the mouse to any corner of the cut to be zoomed in. Press and hold down the left mouse button and drag to the opposite corner of the cut, then release the button - the selected cut will be shown magnified in the main graph. The station stores the last cuts; they can be displayed one by one by icons and or the commands Display - Previous (Next) Zoom. Icon or the command Display - Unzoom displays the chromatogram in the original size. Note: Original size is either the size when the chromatogram fills the whole graph area, or the size specified in the Axes tab of the Graph Properties dialog box by the parameters From To, provided however that the items Range - Fixed are checked. 52

53 User s Guide Chromatogram 6.2 Display Characteristics In view of the many available options for displaying chromatograms, axes, tags and other information, a separate dialog box Graph Properties containing several tabs is devoted to these settings. Open the dialog box by the command Display - Properties from the main menu or Properties from the local menu (to pop up the local menu click with the right mouse button). Parameters are organised in several tabs described below. Global setting It is convenient to set a number of parameters common to other graphs (chromatograms in other channels, the Calibration or Data Acquisition windows); the station therefore offers a global setting for some parameters by the command View - Options in the Channel window. The opened dialog box User's Options offers the tabs Graph Defaults, and Axes devoted to the setting of graphs. If you check the items Use Defaults on synonymous tabs in windows Chromatogram, Calibration and Data Acquisition, the corresponding parameters will be taken over from this global setting. Note: The setting from the channel window and from individual windows is stored in a file; in the unprotected mode the file is EAS.DSK and is common to all channels. See Chapter The Desktop File (EAS.DSK) on page 110 for additional details Chromatogram Description and Display Select the information to be displayed on Graph tab of the Graph Properties window. Fig. 14 The Graph tab of the Graph Properties Window 53

54 EAS32 v.1.4 Chromatogram Description and Format of Displayed Axes The Axes tab of the Graph Properties window enables the user to specify own descriptive labels at both chromatogram axes as well as to decide that the chromatogram will be displayed in fixed rendition (fixed range of the axes). Axes description The thickness and colour of axes are set by the commands Line Width and Color; the commands Title Font, Value Font and Units Font decide on the fonts used for axes description, values and units. Fig. 15 The Axes tab of the Graph Properties window Axis format: In the right section select the actual format of the axes, for each axis separately. You may cover the axis by crossing out the command Show. Enter the actual title in the item Title. It is possible to shift the axes - Offset or change their scale - Scale. Using the item Units select display of the axes in the required units. The units are displayed only on the axes of the main graph and not in the results tables, and thus have a purely informative meaning. 54

55 User s Guide Chromatogram How to set actual units on the axis In chromatographic practice it occurs that instead of voltage it is better to display the absorbency units (AU). The procedure for setting is as follows: 1. Open the window Graph Properties on the Axes tab. 2. In the pull down menu of the axes, select Voltage Axis and check whether the item Show underneath has been marked. 3. Select the name of the axis in the item Title, e.g. Absorbency units and the abbreviation of units AU in the item Units Type. 4. In the item Units Type select whether you wish to display the axis with a fixed scope of values Fixed, or whether the station is to automatically convert the values to decimal multiples expressed by the prefix before the name of the unit (mili, micro, kilo, etc.) item Auto. 5. Ascertain what voltage corresponds to 1 AU and enter its inverse value into the item Scale. e.g.: If 1 AU corresponds to a voltage of 2V, enter into the Scale item the value 0.5. Fixed rendition Fixed rendition means that both axes will have a fixed range regardless of the actual chromatogram size. The range is given by the parameters From To in the Range section always in the base units (millivolts and minutes). The specified values apply only if the Fixed item is checked Descriptive Labels and Lines The main graph can contain descriptive labels and lines. Both can be situated in the graph area or anchored to the current chromatogram. Descriptive label location Invoke the command Chromatogram - Create Label - Text from the menu or from the local menu to display a special cursor. Move the cursor to the location of the future descriptive label. Click to open the Text Label dialog box where you can enter the text of the label in the Text item, select font by the Font item, and decide on orientation (from 0 to 360 ) by the Orientation command. 55

56 EAS32 v.1.4 Chromatogram Anchoring to chromatogram To anchor a label to the active chromatogram check the Assign to Active Chromatogram parameter. Invoke the command Anchor (Text Alignment) to decide on the point to be "fixed" to the nearest chromatogram point or to the graph border. Line location Invoke the command Create Label - Line from the main Chromatogram window or from the local menu to display a special cursor. Move the cursor to the location of the future line beginning, hold down the left mouse button to draw a line, and release the button once the end point is reached - the dialog box Line Label opens where you can specify in items Arrows, Line Style and Line Width the type of line end, the colour and type of the line, and its thickness. Note: Lines with line thickness in excess of one pixel will be always shown as continuous. You may anchor the line to the active chromatogram by checking the parameter Assign to Active Chromatogram. Modification of descriptive labels and lines A label or line previously created can be modified anytime by double-clicking it with the left mouse button or clicking it with the right mouse button. Labels and lines can be also moved or resized by means of the "handles" (similar to those used by standard drawing applications), invoked by clicking any point on the label or line. Hold down the left mouse button and drag to move the object, then release the mouse button to drop it at the desired location. The object size can be changed similarly by means of the relevant handles. When a label height is changed the font size is adapted accordingly. Deleting labels and lines Delete the selected label or line (that with the handles shown) by the Delete key or the command Remove Labels - Selected from the Chromatogram menu or the local menu. To delete all labels and lines from a graph invoke the command Remove Labels - From Workplace. To delete all labels and lines from an active chromatogram invoke the command Remove Labels - From Active Chromatogram. To delete all labels and lines invoke the command Remove Labels - Remove All. Note: Labels and lines anchored to an active chromatogram are stored with that chromatogram. 56

57 User s Guide Chromatogram Labels and lines attached to the graph area are a part of the desktop file - extension DSK. Print of descriptions and lines In the print style in the window Report Setup Chromatogram you may set whether descriptions and lines are to be linked to the chromatogram or are to be simply printed with the area. This may be used amongst other matters also for your working symbols and notes which should not be a component of the print output. 6.3 Operations Involving Several Chromatograms - Overlay In the OVERLAY mode several chromatograms can be displayed simultaneously. OVERLAY mode enable To enable the OVERLAY mode use the command Overlay from the File menu or icon, or double-click the OVERLAY mode displayed to the right on the status bar. Active OVERLAY mode is indicated by the indented icon, coloured icons, and on the Overlay toolbar, and the inscription OVERLAY to the right on the status bar. Displaying several chromatograms Up to 100 overlaid chromatograms can be displayed simultaneously by either the command File - Open or by clicking icon. Chromatograms are displayed by invoking repeatedly the command Open or by selecting several chromatograms in the Open Chromatogram dialog box. Each chromatogram is displayed in dedicated colour, and icons are assigned to the first eight chromatograms displayed. Refer to the Chromatogram menu for a comprehensive list of all displayed chromatograms. Note: To select several files hold down the Ctrl key while clicking with the left mouse button (an additional click cancels the selection). To select a contiguous sequence of files click the first file to be selected with the left mouse button, hold down Shift and click the last file to be selected. 57

58 EAS32 v.1.4 Chromatogram Current chromatogram One chromatogram - usually that last displayed - is current, and all displayed information and commands refer only to that chromatogram. The name of the current chromatogram is shown in the title bar of the Chromatogram window and in the headers of all tables, and the icon with the corresponding colour is indented ( we have marked it with the circle in the following picture). To select another chromatogram as current click the icon with the corresponding colour or select the chromatogram from the list in the Chromatogram menu. Fig. 16 The Overlay Toolbar Changing chromatogram colour Changing the colour of any of the first eight chromatograms is simple: click the icon of the chromatogram whose colour you wish to change and then an empty icon with the required colour. The colour of the icons and the chromatogram will change accordingly. Closing a chromatogram To close the current chromatogram invoke the command Close from the File menu or click icon. Invoke the command Close All to close all displayed chromatograms. Disabling the OVERLAY mode To disable the OVERLAY mode click the Overlay command from the File menu or the indented icon. The station may invite you to save modified chromatograms about to be closed. When the OVERLAY mode is disabled only the current chromatogram will be displayed in the graph Resizing and Relocating Chromatograms The current chromatogram can be resized and moved independently in the horizontal and vertical directions. In this manner you can easily "match" two chromatograms, e.g., to subtract the solvent peak or to modify a chromatogram with erroneous retention times. These modifications are effected by means of the commands Move and Scale from the Chromatogram Overlay submenu or by means of icons and. 58

59 User s Guide Chromatogram Modification procedure Invoke the command, place the mouse cursor at any part of the main graph, hold down the mouse button and slowly drag in the desired direction - the chromatogram will move and change its size. Once the requested location or size has been reached, release the mouse button. Displaying/changing values The values by which the chromatogram has been changed are shown in Line Charts tab of the Graph Properties window in parameters Offset X (Y) and Multiplier X (Y). The chromatogram can be modified directly by changing the values of these parameters. Invoke the command Origin to return to the original size and location. Saving the effected changes The chromatogram modified as described above becomes a mere graphical curve, i.e., the baseline and the peak descriptions disappear. To create a full-bodied chromatogram from the curve invoke the command Overlay - Operation - Copy, followed by the command File - Save (see Chapter Mathematical Operations) Mathematical Operations Some basic mathematical operation over chromatograms can be performed by the command Overlay - Operation from the Chromatogram menu by clicking icon. In the opened dialog box Mathematical Operations click the colour of the first chromatogram in the left column Chrom A, select the required operation in the middle column Operation, and select the second chromatogram, if any, in the right column Chrom B. Fig. 17 The Mathematical Operations Dialog Box 59

60 EAS32 v.1.4 Chromatogram The colour of the resulting curve is shown beneath the inscription Result after the equal sign. The result is automatically integrated and evaluated according to the method selected in the Channel window. The method associated with the original chromatogram is never used when a copy is created or differentiation performed by the commands Copy and Differentiate, respectively; it is only used for a copy created by the command File - Save As! Any chromatogram created or modified in this way is labelled in the report by the inscription "Mathematically reprocessed", The created chromatogram is not saved and will be cancelled when the window is closed. If you want to preserve it, save it by the command Save or Save As Three-dimensional View of Chromatograms To display chromatograms in 3-D view invoke the command 3D View, to cancel the view invoke Clear 3D, both from the Chromatogram - Overlay submenu. The two straight lines displayed by the 3D View command are used to set up the angle and depth of the three-dimensional view. Click to display all chromatograms regularly displaced along the selected line. The displacement is governed by the items Offset X and Offset Y in the Line Charts tab of the Graph Properties window. The command Clear 3D zeroes the above offsets and, accordingly, displays all chromatograms at their original positions. The original positions of individual chromatograms are set by the Origin command, again from the Line Chart tab of the Graph Properties window. 6.4 Chromatogram Modifications By default each chromatogram is integrated according to the Peak Width and Threshold parameters and other parameters shown in the right-hand side of the Integration tab in section Parameters. The corresponding baseline can be however modified in any way to conform as closely as possible to your requirements. To this end the station offers a number of commands, all in the Chromatogram submenu. The effected commands are stored in the integration table, and the latter can be subsequently modified or used for another chromatogram. Note: The station automatically checks for instances of baseline crossing. After each operation the baseline is corrected to prevent baseline crossing (the station either shifts the peak beginning or peak end to the nearest possible point or refuses 60

61 User s Guide Chromatogram to perform the requested operation). Accordingly, if the peak beginning or end has been shifted manually, it may happen that the station refuses to place the point at the requested position, since it detected that the baseline would intersect the signal, and places the point at the nearest possible location. The method contains also some global parameters that affect the peaks included in integration - Rejection, and baseline course - Separation. Fig. 18 The Right-hand Side of the Integration tab Order and hierarchy of parameters and commands affecting integration 1. Peak detection using the parameters Peak Width and Threshold starts from the time indicated in the parameter Integration Start. 2. An attempt then follows to draw the baseline to the valley according to the ValleyToValley Slope parameter, followed by automatic tangential separation according to the remaining two parameters of the Separation section. 3. The Integration Table is applied. 4. A check of possible baseline/signal intersection is performed. 5. Finally, all peaks that fail to satisfy at least one of the parameters specified in the Rejections section are excluded. Note: It follows from the above that separation parameters are not applied, e.g., to manually added peaks and, on the other hand, their effect might be suppressed, e.g., by the commands Together and Valley specified in the integration table. Major commands and global parameters are explained and their effect illustrated in the following chapters. 61

62 EAS32 v.1.4 Chromatogram General modification procedure Click the corresponding icon on the toolbars Baseline and Peak or from the Chromatogram - Baseline (Peak) menu to initiate the manual chromatogram modification procedure. The mouse cursor is displayed approximately at the centre of the active chromatogram as a vertical line with an arrow pointing to the signal level. Point the mouse cursor to the place where you wish to effect the requested operation and click with the left mouse button. If an interval operation is involved, a second vertical line appears and the second point can be selected similarly. The user can cancel the operation anytime by the right mouse button or the Esc key Baseline Modifications The Baseline submenu and the corresponding toolbar contain commands used for baseline modification operations and to eliminate peaks from integration. Fig. 19 The Baseline Toolbar Validity Interval The interval of validity is delimited by two vertical lines. When the command is invoked the left vertical line appears; drag it to the required position by the mouse and confirm by clicking with the left mouse button. Place the right vertical line to the required position in the same way - its confirmation invokes the command. The command can be abandoned anytime by clicking with the right mouse button or pressing Esc. Operations involving peaks will be only performed with peaks completely enclosed between the two vertical lines. Records of modifications All accomplished modifications are stored in the integration table, where any operation can be subsequently cancelled or corrected. An integration table from another chromatogram or another method can be also used (see Chapter Integration Table on page 72 for additional details). 62

63 User s Guide Chromatogram Peak deletion The command Lock or icon excludes all peaks from integration. Only a peak completely contained in the selected interval will be excluded. Baseline through a valley Use the Valley command (or icon ) to force the baseline to all valleys situated between the interval lines. In the event this would result in the baseline intersecting the signal, the station will shift the peak beginning or end automatically to prevent it. This is the inverse operation to Together. Baseline between separating verticals The Together command (or icon ) introduces separating verticals to all valleys situated between the interval lines. This is the inverse operation to the Valley command. 63

64 EAS32 v.1.4 Chromatogram Horizontal baseline The command Forw. Horizontal (or icon ) forces a horizontal baseline from the beginning of the first peak inside the selected interval beneath all peaks inside the interval. The command Back. Horizontal (or icon ) draws a horizontal baseline from the end of the last peak inside the selected interval beneath all peaks inside the interval. Note: Should the horizontal baseline intersect the signal inside a peak, no forced horizontal baseline is possible for that peak. Tangential separation The command Front Tangent (or icon ) cuts off all peaks (riders) situated on the leading edge of the first peak after the selected interval. Place the left interval line before the peak you still want to cut off, the right interval line inside the peak starting from which the cut-off is to be effective. The command Tail Tangent (or icon ) cuts off all peaks (riders) situated on the tailing edge of the last peak before the selected interval. Place the left interval line inside 64

65 User s Guide Chromatogram the peak that carries the tailing riders to be cut off, the right interval line behind the end of the last peak to be so cut off. Interchange positive/negative The command Clamp Neg. (or icon ) interchanges all apexes inside the interval for valleys and vice versa, thus creating positive peaks from a group of negative ones. Declaring negative peaks to be disturbances The command Cut Neg. (or icon ) eliminates from integration negative peaks that are in fact disturbances. Neither the beginning nor the end of the preceding positive peak will be affected. 65

66 EAS32 v.1.4 Chromatogram Declaring peaks to be baseline The command Reject Negative (or icon ) excludes from integration negative peaks without declaring them to be disturbances. The beginning of the subsequent peak or the end of the preceding peak (if any) is shifted to the original apex of the negative peak Peak Modifications The Peak submenu and the corresponding toolbar contain operations with peaks. Fig. 20 The Peak Toolbar The positions of peak beginnings or ends as well as of valleys or the separating vertical lines can be changed. Invoke the command, move the cursor to the required location and click to confirm. The command may be abandoned anytime by clicking with the right mouse button or hitting the Esc key. Since only a single peak beginning or end can be shifted to any place selected on the chromatogram without shifting the apex, it is unnecessary to specify the peak to which the operation refers. Note: The preceding sentence does not apply to tangential separations where, if the required location lies inside the main peak, modification of the tangentially separated peaks is given preference. 66

67 User s Guide Chromatogram Shifting peak beginning The command Start (or icon ) defines a new position of the peak beginning. In the event the peak beginning is shifted before the end of the preceding peak, those peaks will be separated by a vertical line and its position will coincide with the new peak beginning. If the peaks are separated by a vertical line or connected at the valley and the peak beginning is shifted closer to the apex, the peaks will be separated. Shifting peak end The command End (or icon ) defines a new position of a peak end. In the event the peak end is shifted behind the beginning of the following peak, those peaks will be separated by a vertical line and its position will coincide with the new peak end. If the peaks are separated by a vertical line or connected at the valley and the peak end is shifted closer to the apex, the peaks will be separated. 67

68 EAS32 v.1.4 Chromatogram Change valley or the vertical separating line The command Both (or icon ) defines a new position of a valley or the vertical line separating the peaks. The common point can be shifted to any location between the apexes of the two peaks. Should the resulting baseline intersect the signal, the peaks will be separated. Adding new peaks The beginning/end of a new peak is given by the interval lines, the apex is determined automatically at a location between the maximum and minimum (depending on peak orientation) between the peak beginning and end. A new peak cannot be added in the area of tangentially separated peaks. Adding a positive peak The command Add Positive (or icon ) creates a new positive peak. If the beginning or end of the created peak falls inside a neighbouring peak, the peaks will be separated by a vertical line situated at the beginning or end of the created peak, as the case may be. 68

69 User s Guide Chromatogram Adding a negative peak The command Add Negative (or icon ) creates a new negative peak. If the beginning or end of the created peak falls inside a neighbouring peak, the peaks will be separated by a vertical line situated at the beginning or end of the created peak, as the case may be. Manual apex determination Owing to the automatic determination of peak apex the following procedure must be used to place the apex at a specific location (especially when defining monotonic peaks): First cancel the peak with incorrectly assigned apex by the command Lock and add a new peak by the command Add Positive or Add Negative as required so that its beginning or end (depending on the position of the maximum of a positive or minimum of a negative peak) coincides with the position of the intended apex. Finally shift the beginning or end to the appropriate location by the commands Start or End. Solvent peak The command Solvent Peak (or icon ) designates a peak to be a solvent peak. The designation is indicated by letter "S" before the peak number in the graph. Solvent peaks are neither included in integration nor listed in the Result Table. More than one peak may be designated solvent peak. Record of accomplished modifications All accomplished operations are stored in the integration table and, accordingly, any operation can be cancelled or corrected. Integration table from another chromatogram or another method can be also used (see Chapter Integration Table for additional details). 69

70 EAS32 v.1.4 Chromatogram Selection of Conditions Restricting Integration The Rejection section in the Integration tab can be used to exclude from integration peaks that do not satisfy certain criteria. All peaks whose Area, Height or width at half height - Half Width are smaller than or equal to the specified values are excluded from integration, in other words, are neither displayed in the chromatogram nor included in the Result Table. In this manner (in particular by the Height parameter) you can get rid of small, insignificant peaks that confuse the results without affecting the baseline under the remaining peaks Separation Parameters The Separation section in the Integration tab can be used to change the baseline below not separated peaks. See the Reference Manual for additional information Integration Table Integration table contains a list of "manual" chromatogram modifications, i.e., the commands from the Baseline and Peak submenus. Each operation is identified by its name and the range of validity. For commands with range of validity defined by an interval (Lock, Add Positive, ) the integration table contains the limits of the validity interval. For commands defined by a single point (Start, End and Both) the integration table contains the retention time of the peak involved and the new distance relative to the retention time. Note: This approach to a certain extent removes potential inaccuracies originating from differences in retention times determined in various analyses. Integration table for new chromatograms: You can prepare the integration table for new chromatograms before measuring the chromatogram in the window Method Setup Integration, which you may open with the command Method - Integration in the channel window. 70

71 User s Guide Chromatogram Use of integration table from other method for measurement of new chromatograms: Display the integration table in the channel window by using the command Method - Integration. The local menu here contains the command Copy From. After selecting, then select the corresponding chromatogram or method. This operation is irreversible. Use of method or only integration table from another chromatogram: The Method menu in the window Chromatogram Integration contains amongst other elements two sets of three commands for copying methods or only integration tables alone from other chromatograms: Copy (Integration Table) from Chromatogram Use of method (or only integration table) from another chromatogram. Note: It is necessary to differentiate copying of an entire method from copying only an integration table. 71

72 EAS32 v.1.4 Calculations and Calibration 7 Calculations and Calibration The chromatographic station EAS32 offers several types of calibration and noncalibration calculations and, moreover, can assess the measurement quality by means of calculated peak and column parameters. Extensive mixtures can be calibrated at up to twenty concentration levels. The EAS32 incorporates the reference peak method for reliable identification of calibrated compounds. 7.1 Result Table The Result Table is in the Results tab, which can be opened or hidden by the command Results - Result Table, and contains always the topical values referring to the current chromatogram; any changes are immediately reflected in the table. Fig. 21 Table of Uncalibrated Results Results are displayed in dependence on the setting of parameters in the right-hand side of the tab, and the arrangement of columns is also governed by the Setup Columns command. In the following text we describe the default layout of the table, which is the result of the command Restore Default Columns. In addition to the serial peak number, the retention time and width-at-half-height of all peaks, the absolute calibration values and percentages, names and types of compounds are shown for uncalibrated results. If you set a calibration file by the Set command, the absolute calibrated values and percentages, the names and types of compounds will be also shown. 72

73 User s Guide Calculations and Calibration 7.2 Types of Integration Calculations Generally speaking the result of a chromatographic analysis, representing the weight of a compound, is either the area or height of the corresponding chromatographic peak. The same weights of different compounds however elicit different detector response. To take this fact into account in calculating the weight, one must know the detector sensitivity under the given measurement conditions, in other words, the calibration curve of the compound in question, representing the response as a function of the weight of the compound. Calibration curves are stored in a calibration file; the latter can contain an arbitrary number of calibration curves corresponding to compounds present in a certain mixture. The Calibration window is earmarked for creation and modification of calibration curves. Calibration calculations respect the differences in detector sensitivity vis-à-vis individual compounds at various concentrations. Concurrent calibration of several compounds from a single injection may register potential interactions between the calibration compounds as well. If a tentative evaluation of the weight of the compound is sufficient for your purposes, use the uncalibrated calculation where no calibration curve is required. Calculation of percentages The ratio of parameters Weight and Dilution from the chromatogram header specifies the overall weight of components in calculation of percentages. If the parameter is left blank, the total weight is taken as the sum of calibrated weights of all compounds including unidentified compounds. Note: If the sum total of percentages of all evaluated compounds exceeds 200 %, individual percentages are not stated and the column Weight[%] is filled up with question marks. Calibration curve To enable calibration curve compilation the calibration file must contain at least one calibration level at which the given compound has been calibrated, otherwise the response coefficient from the main calibration table is used in the calculation instead of reading the value off the (non-existent) calibration curve. The Result Table then contains, at the corresponding compound in column Peak Type, the message Free as an indicator that the above coefficient has been used. This however does not apply to noncalibrated calculations. 73

74 EAS32 v.1.4 Calculations and Calibration Non-calibrated Calculations - Uncal Without a calibration file the weights can be only expressed as percentages of the overall area or height of all chromatograms. The two types of non-calibrated calculations assume that the detector sensitivity is the same for all compounds and that all compounds present in the injected sample were actually detected. Since the first assumption is very rarely met, non-calibrated calculations are used primarily to obtain a preliminary, semi-quantitative result. Note: Non-calibrated calculations are used by default also in instances where calibrated calculation has been selected but no calibrated compound was identified in the chromatogram or the requested calibration file was not found. The type of calculation actually employed is always indicated in the header of the table of integration results - Results External Standard Method - ESTD The calculation by the external standard method respects differences in detector sensitivity to various compounds, and provides weights of individual compounds and their percentages in the injected sample. The drawback of the external standard method is its extreme sensitivity to both the injected weight and the measuring conditions. To eliminate possible errors it is thus mandatory to maintain high precision of injected weights (autosamplers) and frequently rebuild the calibration files so that they reflect potential changes in characteristics of the detector and the chromatographic system as a whole. 7.3 Description of the Calibration File The calibration file contains the data items needed for compilation of calibration curves. In addition to the table of weights and responses other data are listed for each compound, like the compound name, the retention time, the number of calibrations, etc. The data contained in the calibration file are presented in two tables. The main calibration table lists data of all compounds that are common to all calibration levels, while the compound tables contain data referring to individual compounds at all levels, including the calibration curve. Note: All data items stored at several places are always updated after each change. Naturally, when a modified calibration file is used to process the displayed chromatogram, the changes are immediately taken into account. 74

75 User s Guide Calculations and Calibration Main Calibration Table The main calibration table is available from General tab of the Calibration window. Fig. 22 The Main Calibration Table Open the window by the Window - Calibration command or icon. The table summarises data about all compounds, common to all levels: in addition to the name of the compound and the retention time also the two identification time windows, the compound type - Peak Type, the global response coefficient - Response Factor - used in free calibration and, in column labelled Level X, data of the current calibration level. The table can be displayed above or below the calibration standard graph - see the View - Table Up command Compound Calibration Tables Calibration tables of individual compounds are available - together with the calibration curves and other parameters - from tabs of individual compounds in the Calibration window. Display the relevant tab by double-clicking its name or the first grey field of the corresponding line in the main calibration table. The table lists values for all levels of the relevant compound: level number in column Lvl, responses in column Response, and weights at individual levels in column Weight. The Response Factor column contains the level-specific response factors, equal to the weight divided into the response at that level. 75

76 EAS32 v.1.4 Calculations and Calibration Note: The level-specific Response Factor is merely an indicative value. In the event free calibration is to be employed the global response factor taken from the main calibration table is always used. Fig. 23 The Compound Calibration Table Display areas or heights? The base (either the peak area or peak height) of the displayed response, of the level response factor, of the correlation equation and of the displayed curve is determined by the Response Base parameter. Optional validity of points: Individual points may be temporarily left out or on the other hand included into the calculation of the calibration curve. A point is omitted from the calculation by cancelling the indication in the column Used in the calibration table of substance. An invalid point is illustrated in the graph by a circle. Note: An invalid point in the calibration curve behaves as if it were not present, thus if you for example indicate it as an invalid point at the end of the curve the scope of the axes changes after display of the curve and a circle is shown outside of the displayed area of the graph. Only the circles can be seen of those invalid points behind which is still at least one valid point (cross). 76

77 User s Guide Calculations and Calibration Calibration Curve The calibration curve is shown together with the compound calibration table. The curve is plotted as the dependence of response on weight. The curve is not displayed, though, if the correlation equation cannot be compiled for the selected type of correlation. Each calibrated level is shown by a cross in the graph; points for recalibrated values at some levels, if any, are shown as asterisks (the number of calibrations on one level is apparent from the main calibration table column Rec. No.) Fig. 24 Example of Calibration Curve Any part of the curve can be zoomed in by the left mouse button; return to any of the previous magnifications or to the original cut by the commands Previous Zoom (icon ), Next Zoom (icon ) or Unzoom (icon ), respectively, or display a grid by the synonymous command. All the above graphical functions are the same as in the Chromatogram or Data Acquisition windows (e.g. Chapter Displaying a Chromatogram on page 53 ). Type of curve fit equation The type of correlation is defined in item Curve Fit Type; item Origin decides on how the co-ordinates of the origin will be handled: the Ignore Origin option ignores the origin while with the Compute with Origin option the origin is considered to be one of calibration points; this means that the resulting curve is influenced by, but need not pass through, the origin; the Curve Passes Through Origin option forces the correlation curve always to pass through the origin. 77

78 EAS32 v.1.4 Calculations and Calibration Correlation equation Item Equation shows the plot of the calibration curve and the corresponding correlation equation (except for the Point to Point (segmented line) type of fit and Free Calibration), with X representing the weight and Y the response. The curve and the corresponding equation need not be always displayed and, especially in instances of third-order polynomial correlation, the algorithm used to calculate the coefficients of the equation may fail for some extreme values of weights and responses. To remedy such situations use another curve fit type or normalise the entered values. Tightness of fit The Correlation Factor item contains the calculated correlation coefficient - a number from the interval <0; 1>, characterising the tightness of fit. Note: In the event the degree of the correlation equation is the same as the number of calibration levels (for the Ignore Origin option already for a degree less by one) the correlation coefficient will be 1 since such a curve will pass through all points. 7.4 How to Create Calibration The following chapters describe the procedure used to create and modify calibration files. The station provides for both automatic and manual calibration of individual peaks, or jointly for all compounds (peaks). 1. Create the calibration file Create a new calibration file by the command File - New from the Calibration window. 2. Display a calibration standard Unless you intend to create an "artificial" calibration you must display in the calibration window the relevant chromatogram that represents a calibration standard at a certain level - invoke the command Open Standard or click icon to display a list of all chromatograms from the calibration subdirectory of the current project. Use icon to display chromatograms from the data subdirectory. 3. Selection of calibration parameters Check the toolbar whether is set. The number indicates the calibration level, Automatic means that data for individual compounds will be transferred automatically to the calibration table. 78

79 User s Guide Calculations and Calibration 4. Transfer data for individual compounds to the calibration table Use any of the following commands from the Calibration submenu or the corresponding icon to transfer data of compounds contained in the calibration standard to the calibration file: Add All - if the calibration table is empty, all peaks from the chromatogram will be calibrated. Add Peak - Only the peak selected by the cursor will be calibrated. 5. Supply known weights and other data items Enter all known weights for all compounds in column Weight below the calibration level - Level 1. Update the compound name in column Compound Name. 6. How to display and check the calibration curve? Click the tab with the name of compound whose calibration curve you want to see; the calibration table, the calibration curve and parameters of that compound will be displayed. 7. How to redisplay the main calibration table and the standard? Click the General tab. 8. Saving the calibration file To save the calibration file invoke the File - Save command. 9. How to use the calibration file? Open the Chromatogram window by the command Window - Chromatogram or by icon. Open the chromatogram to be evaluated by the calibration just created. Set the created calibration file by the Set command in the right-hand side of the Results tab. If all went well the Weight column of the result table should now contain the corrected weights for all calibrated and identified peaks. 7.5 Creating Additional Concentration Levels Once the first calibration level has been created, continue to calibrate at the next levels. Any number of compounds can be calibrated at each level regardless of whether or not 79

80 EAS32 v.1.4 Calculations and Calibration they have been calibrated at the lower levels. Moreover, compounds from different calibration standards can be calibrated at a single level. 1. How to set another calibration level? Each time a calibration file is read in the first calibration level is always selected. Use the arrow in the toolbar or command Calibration Set Level to change the calibration level. If the Automatic regime is enabled the current calibration level is used. To prevent the program from selecting calibration levels automatically, enable the regime Manual on the toolbar. See Chapter Manual Calibration for additional details. 2. Reading in another calibration standard Invoke the command Open Standard to select and display a calibration standard corresponding to the preset calibration level. 3. Transferring compound data to the calibration table Invoke any of the following commands: Add All - If the calibration table contains any compounds calibrated at the previous level, only those will be identified and potentially calibrated. Add Peak - Only the peak selected by cursor will be calibrated. 4. Supply known weights Enter all known weights for all compounds in column Weight below the calibration level - Level x. 5. Repeat items 2 to 4 for all calibration levels required Note: Note: Thanks to the character of the EAS32 any change in the calibration file is immediately reflected in the result tables of all chromatograms that use it. In other words, it is not necessary to save the changes by the command File - Save to update the result table. 6. How are peaks assigned to compounds in the calibration table? The retention time of each peak you wish to calibrate by the command Add Peak will be compared with the identification windows of already calibrated compounds. If identified with any of them (i.e., if the retention time lies within the interval < Ret. Time - 80

81 User s Guide Calculations and Calibration Left Window; Ret. Time + Right Window>, its retention time is added at the next level. Otherwise a new calibrated compound is established. If the calibration proceeds under the command Add All, only identified compounds will be calibrated in the above manner, in other words, you cannot add new compounds in this manner. 7. Updating retention times of compounds The retention time of any compound can be changed manually anytime in the Retention Time column. The retention time is also updated automatically during each new calibration of the compound in question. The updated retention time is taken as the arithmetic mean of all values established by existing calibrations of that compound. The automatic update can be switched off by unchecking the item Update Retention Time in the Calibration Options dialog box. 7.6 Manual Calibration If you wish to acquire full control over the calibration process, reset the parameter Automatic to Manual in the toolbar. Fig. 25 The Calibrate Peak Dialog Box After using certain of the commands Add All, Add Peak in the manual regime, the dialogue window Calibrate Peak is displayed for each compound, in which you may immediately amend and enter all data on the calibrated compound. For the Add All command you can then skip the calibration of the currently displayed compound by the Cancel button. The Abort button terminates the whole calibration process. The weight of the calibrated compound is entered in item Weight in the displayed window. 81

82 EAS32 v.1.4 Calculations and Calibration 7.7 Calibration File Modification All data in calibration files can be changed unless dimmed. Manual modification of responses Item Enable Response Value Change in the dialog box Calibration Options must be checked prior to any modification of the values of responses. The number of calibrations is reset to 1 when a response is changed manually. Deleting a compound To delete a compound invoke the command Calibration - Delete Compound or click icon. "Artificial" calibration There are no obstacles to creating a calibration curve in the absence of a calibration standard, simply from the known response factor or response. Add a new compound to the main calibration table simply by filling in any cell in the last free line. Once the entry is confirmed by Enter, the remaining data will be supplemented automatically and can be subsequently modified Default Parameter Settings Invoke the command Calibration - Options to open the Calibration Options dialog box; then click the tab of the Defaults tab. Fig. 26 The Calibration Options - Defaults Dialog Box 82

83 User s Guide Calculations and Calibration Here you can select the default values of some selected parameters for new compounds in the current calibration file Response Factor and Free Calibration The calibration file lists two response factors for each compound. Level-specific response factor For each calibrated level the compound table lists the response factors in the synonymous item. The factor is calculated as weight divided into response at a given level and is only indicative. Global response factor The main calibration table contains the global response factor - Response Factor, calculated as the reciprocal value of the slope of the straight line passing through the calibration points. Free calibration In free calibration - Free Calibration - the global response factor is used instead of values calculated from the calibration equation. The user can specify its value also when the compound in question has not been calibrated. The value entered manually is not updated when calibration points change. Should you wish to display the original value, enter zero or use the command Use Original or Factor from the local menu. Free calibration can be selected manually on the tab of the selected compound in item Curve Fit Type. Free calibration is applied automatically to compounds for which a calibration equation cannot be established, and its use is indicated by the inscription Free in column Peak Type of the Result table in the Chromatogram window. 7.8 Peak Identification in Calibrated Calculations One may reasonably expect that compounds contained in a calibration file will be identified with peaks in the measured chromatogram. To this end the user specifies identification windows relative to the relevant retention time for each compound in the calibration file. If a peak exists in the chromatogram whose retention time falls within the identification window, it is assigned to the corresponding compound from the calibration file. In calibrated calculations the calibration curve for that particular compound is subsequently used for the peak thus identified. If there are several peaks inside the iden- 83

84 EAS32 v.1.4 Calculations and Calibration tification widow, the peak whose retention time is the closest to the retention time of the calibrated compound will be identified. Peak identification is used not only in calibrated calculations but in multilevel calibration as well. Chapter explains how the station resolves possible instances of overlapping identification windows "Reference Peak" Method The process of peak identification may be sometimes complicated by changes in retention times brought about by changes of measurement conditions, use of different methods, changes in characteristics of the chromatographic system, or simply by random error. The method of reference peaks eliminates a substantial part of these complications, resulting in reliable identification of peaks in the chromatogram. Compounds marked as reference in the calibration file are identified first. The difference between the retention time of a reference compound and that of the corresponding reference peak in the chromatogram is then used to correct the retention times of the other (non-reference) compounds listed in the calibration file. The non-reference compounds are then identified according to the retention times so corrected. The reference peak method is applied automatically if at least one compound in the calibration file is marked as reference, either in the main calibration table, in the Peak Type item, or in the tab of the relevant compound in item Compound Type by selecting Refer. If several peaks fall within the identification window of the reference peak, the largest of them (according to peak area or peak height depending on the setting of item Response Base) will be identified. A reference peak originally not identified in the chromatogram cannot be subsequently identified inside the newly calculated intervals. 84

85 User s Guide Calculations and Calibration The method used to correct the retention times of non-reference peak is apparent from the following Figure: Fig. 27 Scheme of the Reference Peak Method The difference between the actual retention times of reference peaks, Ti, and the retention times listed in the calibration file, T'i, is used to determine, by linear interpolation (linear extrapolation for non-reference peaks past the last reference peak), the correction to be applied to retention times of non-reference peaks. The following relations apply to the i-th peak in individual segments between the reference peaks (A, B, C, etc.): In segment A (before the first reference peak) T 1 Tc = i To i In segment B (between two reference peaks) T 2 T 1 Tci = T 1+ ( Toi T1) T 2 T1 In segment C (behind the last reference peak) i To i T1 Tc = To i - original retention time of peak i Tc i - corrected retention time of peak i T' i - retention time of the reference peak in the calibration file T i - retention time of the reference peak in the sample. Identification windows relative to the retention times so corrected are used to identify non-reference peaks. The method used to resolve overlapping identification windows construed in the above manner is described in the following chapter. The reference peak T 2 T 2 85

86 EAS32 v.1.4 Calculations and Calibration method will provide the most reliable results if reference peaks are distributed fairly uniformly across the entire chromatogram. The calculated, corrected retention times of non-reference peaks apply only to identification during calibration or calibrated calculations, and are not substituted for retention times of compounds in the calibration file or the sample. Updating retention times of compounds included in the calibration file is described in Chapter Resolving Instances of Overlapping Identification Windows It is not necessary to monitor whether or not the neighbouring identification windows overlap. Prior to the identification process proper the EAS32 checks the identification windows of individual compounds and resolves instances of overlap by curtailing some identification windows. The employed method is elucidated in the following Figure. Fig. 28 Resolving Overlapping Identification Windows 86

87 User s Guide Calculations and Calibration The corrected identification windows remain in effect only during the identification process and do not replace the windows specified in the calibration file Updating Retention Times Retention times of compounds tend to shift with time and, accordingly, difficulties with identification of calibrated compounds in the chromatogram may be sometimes encountered. To avoid this problem the EAS32 provides for automatic updating of retention times of calibrated compounds in each subsequent calibration by registering potential shifts of retention times; this process results in reliable identification of calibrated compounds. Enable the automatic updating by checking item Update Retention Time in the Calibration Options dialog box. The retention times listed in the calibration file are updated as arithmetic mean of the new time and all times established in previous calibrations of the given compound. 87

88 EAS32 v.1.4 Reports 8 Reports The EAS32 provides for an arbitrary part of the obtained results to be printed at any printer registered in the Windows environment. 8.1 Reporting Procedure 1. Where to initiate a report? The EAS32 enables reports to be printed from many places. The menus of most windows contain the command Report Setup, icon or the Report button. Always select the window that contains the data you wish to include in the report. Different report styles are preset at different places, but can be modified or replaced at will. Fig. 29 The Report Setup Dialog Box 2. Selecting a printer Check and, if need be, select a printer by the Printer command, including setting print quality and size. 3. What will be printed? Look through all tabs in the left-hand side of the Report Setup window and check or change what will be printed. The name of the report style containing these settings is shown in the title bar of the window. 88

89 User s Guide Reports 4. Selecting a different report style In some instances is it more convenient to use a different style instead of modifying a report style. Invoke the Open command to list all report styles stored in the COMMON directory. 5. How will the printout look? Use the Preview command to see what will be actually printed. 6. Printing reports To start printing the report invoke the Print command. The dialog box showing the printer settings is shown first to enable the user to check once more the correct setting, the number of copies printed, the scope of printed pages, etc. 8.2 Report Style Files A report style is in fact a template containing a list of all sections to be printed and the report layout (size and type of font, borders, spaces,...). During the printing process proper the actual values or files are merely inserted in the corresponding sections. Each report style is stored in the corresponding report style file and its contents can be displayed and modified. The adapted report style can be saved under the same or a different name as a new report style. All activities are concentrated in the Report Setup dialog box. Report styles are stored in subdirectory COMMON and are common to all channels and projects. 8.3 Printer Selection Printer selection is the only part of the Report Setup dialog box that is not included in the report style and is shared by the whole station; in other words, it is not possible to set different printers or different print quality at individual parts of the station. The list of available printers, their setup and connection with the computer all constitute a part of the Windows system and the station takes it over in its entirety. Start the selection by the Printer command, select the appropriate printer by the button, which displays a list of all printers registered by the Windows operating system. Click the line containing the requested printer to make it the current printer while the station is running. The Properties command beside the line with the printer name opens a dialog box where the print size and quality and other parameters can be set. The layout of the window differs among individual printers. 89

90 EAS32 v.1.4 Reports 8.4 Report Style Selection The name of the preset report style is shown in the title bar of the Report Setup window. Invoke the command Open to see a list of all available report styles. Click the name of the report style to display some additional data items, like the author's name in the Created By item or a description of the report style in the Description item. Select the style either by the OK button or directly by double-clicking the style name. Alternatively you can enter the name of the style in the Filename item. The command New creates a new, empty style. 8.5 Report Style Modification Each report style is divided into individual segments, and a separate tab is reserved for each such segment to the left of the Report Setup window. Segments preceded by the symbol will be printed. Double-click the tab of the corresponding tab to include or exclude that segment from the report. The symbol corresponds to item Print in the top-left corner of each tab. Most segments are further subdivided. Item On New Page decides on whether the segment will be printed on a new page. Global parameters that apply to all segments The first tab, Page Setup, defines the parameters common to the printout in its entirety. Fig. 30 The Page Setup tab 90

91 User s Guide Reports Use the commands Form Font and Value Font to select the type, size and properties of the employed font: Form Font will be used to print headings, Value Font for the data items proper. The commands in section Margins decide on the page margins, the command Between specifies the space between individual parts of the printout; all parameters are in mm. Heading The second tab, Labor. Header, contains the laboratory header. Up to ten lines of text can be entered and will appear in the header of the first, alternatively of all pages in the printout, according to the setting of item On 1st Page Only. The header can be framed (item Border) and printed on grey background (item Grey Background). Item Number of Lines specifies the number of lines per page. A separate font type, font size and characteristics can be selected for each line by icon, and text alignment by icons,, and. Fig. 31 Example of Laboratory Header The contents of all printed segments outside the Report Header can be specified in more detail in the corresponding tab. The segments Method and Sample Table always comprise several parts of the relevant files. The Results segment enables the user to decide whether the table of integration results or the table of column parameters will be printed. The Calibration segment allows the user to decide whether all or only the current level and the calibrated compound will be printed. Printing the chromatogram Parameters from the Chromatogram segment define the orientation and size of the printed chromatogram. Item Orientation is used to select orientation, location and size of the chromatogram: when Portrait is selected, the chromatogram will be printed across the page (with the time axis parallel to the text). The chromatogram will occupy 91

92 EAS32 v.1.4 Reports the entire page width and, unless a fixed height is specified by the Fixed Height command, the default height will be 2/3 of the page. When Landscape is selected the chromatogram will be printed on a separate page with the time axis along the longer side of the page - use the parameter No. of Pages to print the chromatogram on several pages. Printing in Colour The station can print on colour printers, where chromatograms, descriptive labels and lines are printed in the corresponding colour. 92

93 User s Guide Export 9 Export The EAS32 enables data to be exported to the Clipboard, to a text or database file; moreover, chromatograms can be exported in the vector picture format. 9.1 Exporting a Chromatogram and the Corresponding Results as Text The command Export icon from the Channel window or in the Chromatogram window - is used to export data as text. In the Channel window the command only sets up the mode of automatic export by the Postrun function after analysis termination or during batch processing by the Batch command. The Export command in the Chromatogram window is earmarked for exporting manually the active chromatogram. In both instances the dialog box Export Chromatogram opens. Fig. 32 The Export Chromatogram Dialog Box What can be exported? The Export command enables the user to export data from the Results Table (item Results), from the column parameters table (item Column), and also the chromatogram proper (item Data), all in a user-defined format. When a chromatogram is to be exported the section Data is made available where the user can select export of the time axis (item X Axis) and export of either the chromatogram in its entirety or only the currently displayed part (items All Data or Displayed). 93

94 EAS32 v.1.4 Export In what format is the chromatogram exported? The chromatogram is exported as one or two columns of values separated by a character selected from item Text Format. The Fixed format adds the required number of spaces between values of different length to ensure that they are properly aligned in columns of equal width. The first column contains time in minutes, the second column voltage in mv. The time increment is taken from item Time Step. Each voltage is calculated as arithmetic mean of all voltages within the specified time increment. If zero has been entered, the data will be transferred with the minimum time increment of the chromatogram in question (defined by the sampling rate - Sample Rate). If a value larger that the chromatogram duration is specified as the time increment, the result is a single number representing the voltage averaged over the entire chromatogram. No other data (baseline, marks, retention times, descriptive labels) are exported. In this instance the Headers item inserts headings of individual columns. The Full Format item inserts the chromatogram header at the beginning of the exported text. In what format is the Results Table exported? Values of individual cells of the table are separated by a character taken from the Text Format item. The Fixed format adds the required number of spaces between values of different length to align them into columns of equal width. The Headers item adds headers to individual table columns. Item Fixed Format appends the name, date and time of chromatogram creation to each line of the table. What characters can be used as separators? The characters separating individual columns are selected in item Text Format. If Delimited - Comma, Delimited - Semicolon or Delimited - Tab is selected, the corresponding separator will be inserted between individual data items. To preserve the layout of the table use the Fixed format; this format inserts between individual data items the number of spaces necessary for aligning the values properly (unless however a proportional font is used such as, e.g., System); this selection is suitable for exporting data to text editors in creating final reports. Note: The Fixed format is suitable in the case that you wish to use a document as it is, without further amendments. To classify sections of reports into a document in the text editor, create a separate style, which is to be called e.g. EAS32 report and shall use a disproportionate type font. Then apply this style to passages copied from the report of the station. On the other hand, the Delimited format is suitable if you wish to make further amendments to the report (remove 94

95 User s Guide Export or add lines in tables etc.) Most better text editors contain a function which transfers the text separate with a selected symbol to a full-value table. Destination of exported data The Clipboard item exports the data to the Windows Clipboard. To export to a text file select the File item and specify the filename in item File Name. If you wish to state the name of an existing file, invoke the Browse command to search for the file. Automatically assigned filenames If the File Name item is left blank, data will be transferred to a file of the same name as the chromatogram with extension TXT. Target directories for export If item File Name does not specify a path, the text file will be stored in the same directory as the exported chromatogram. Export to a single file The user can append the exported data at the end of an existing file; to do that check the Append item. Exporting results to a database If item dbase File is checked the data will be exported in the DBF format. In this case only the Results Table will be exported to the file in the full format regardless of whether or not Full Format is checked. 9.2 Exporting Chromatograms as Vector Pictures The command File - Save As Picture exports all displayed chromatograms including all descriptive labels, headers and lines in the vector format WMF (Windows metafile). What is a vector format? In the vector format a chromatogram can be easily incorporated, modified and printed at high quality in a text editor or a graphical application (Word, Excel, Corel Draw, ), since it is exported as a set of individual line segments connecting all its points. What are the advantages of the vector format over the bitmap format? Displaying and printing a vector picture is not restricted by the size of the graph in the Chromatogram window, which otherwise affects the quality and size of data exported 95

96 EAS32 v.1.4 Export as a bitmap. The size of the exported WMF file is roughly the same as that of the chromatogram file. Note: In other words, contrary to a bitmap, a vector picture can be arbitrarily magnified and contracted without any adverse effect on its quality or file size. Select to Clipboard or to File to decide on the export destination - Clipboard or a file with the WMF extension. Detailed procedure of copying a chromatogram to a MS Word document 1. Display the requested chromatogram in the Chromatogram window. 2. Invoke the command File- Save as Picture - to Clipboard. 3. Run MS Word and open the target document. 4. Point by the cursor to the location where the chromatogram picture is to be inserted and select Edit - Insert from the main menu. The inserted picture can be magnified or contracted at will and also modified when double-clicked. 96

97 User s Guide Resolving Problems 10 Resolving Problems This chapter summarises problems most often encountered when working with the EAS32. Problems are listed in chronological order in which they might occur in a typical working procedure Running the Program Message Demo in the main station window The demonstration version of the program was mistakenly run. The demonstration version shows the serial number in window About EAS32 in the form (open the window by the command Help - About EAS32 from the main station window). Message Demo - Wrong S/N in the main station window The serial number of the station listed in dialog box About EAS32 (open the window by the command Help - About EAS32 from the Main station window) differs from the number on the A/D Converter board or in the protective key Sentinel. Message Demo - Missing HW in the main station window The A/D board or the protective key Sentinel either not found or damaged. Board driver INT7 either not installed or inoperative. The driver module of the A/D board or the protective key Sentinel not registered. The A/D board driver not activated. You have either forgotten the password or intend to cancel the passwordprotected mode If the station operates in the protected mode and you have forgotten the password, copy the original file EAS.PSW from the PGM subdirectory of the installation CD to the main station directory, thus transferring the station to the unprotected mode, but at a price: all settings in the dialog box User Accounts will be lost. 97

98 EAS32 v.1.4 Resolving Problems 10.2 Signal Displaying and Measurement Greyed (dimmed) icon accompanied with message Disabled, and command Data Acquisition inoperative No source of detector signal has been assigned to the channel (usually a channel of the A/D converter board). Open the dialog box System Configuration by the command System - Configuration and select the appropriate signal source on Channel1 (2-4) tab in item Acquisition. You are using the Evaluation version. From the main station window invoke the command Help - About EAS32 to display the dialog box About EAS32 and check the serial number. The serial number format of the Evaluation version is 98-XXX. No signal displayed in the Data Acquisition window Probably an incorrect range is selected, so that the current value of the signal lies outside the displayed range. Change the parameters Time and Voltage, preferably to their respective maximum values. Alternately, select the floating range by the command View - Floating Axes to enable the station to select the setting appropriate for displaying the signal. A cut is displayed that lies outside the signal. Cancel the cut by the Unzoom command (Ctrl + * or the icon). You have mistakenly set the same colour for the curve and the background. Invoke Properties from the local menu and check the assignment of colours. Data acquisition terminated prematurely Check whether Enable Autostop is not enabled in Method Setup - Measurement. Data acquisition terminated prematurely or started repeatedly Probably a false start signal occurred in the channel where external signal start is enabled - contact your vendor. Disk full If a requested operation cannot be performed owing to full disk, you have several options: Delete files no longer needed from directories other than EAS32. Archiving and deleting of unnecessary chromatograms or entire projects. With the command Archive from the channel window open the window Backup Create 98

99 User s Guide Resolving Problems Archive. Either archive the older chromatograms on another disk or erase them using the command Delete Selected Files Sample Table The Active Sample Table cannot be run In an Active Sample Table all method files employed must have the Enable Autostop item set. Click in turn with the right mouse button the names of methods in the Sample Table, and display the contents by the command Edit File from the local menu. Click the Measurement tab and check the Enable Autostop item Processing and Displaying Chromatograms A file contains an incorrect chromatogram Check whether the correct project has been selected, since files of identical names can exist in several directories. Maybe you have overwritten the original chromatogram by another one by entering the name of an already existing chromatogram in the Chromatogram File Name and subsequently ignoring the displayed warning against possible overwrite. A completed chromatogram appears clipped from above or from below First of all check whether the chromatogram is not clipped only on the monitor because of the command Properties - Axes - Range - Fixed. If the chromatogram is clipped from above, the detector signal was apparently cut off either owing to a too low range set in item Range in the Acquisition tab of the Method Setup window, or was cut off already in the detector owing to excessive preset range or to injection of an extremely concentrated sample. If negative voltage values are missing from the chromatogram, item Bipolar in Acquisition tab of the Method Setup window was probably unchecked; this item enables measurement of negative detector signal. Excessive noise in the chromatogram You have probably measured with an unsuitable connection between the chromatograph and the computer, e.g., the detector cable of the EAS32 was not shielded. Another rea- 99

100 EAS32 v.1.4 Resolving Problems son might be a defective chromatograph or converter board. Check the chromatograph (e.g. by a recorder), or contact your vendor. Small peaks exhibit a staircase shape or excessive noise You probably do not utilise the full measuring range of the converter. If the voltage of the maximum peak of interest is at least eight times smaller that the voltage range set in the Range item in the Acquisition tab of the Method Setup window, use a lower range. Some peaks are not detected Check whether the limiting integration conditions are not improperly set (item Rejection in the Integration tab of the Chromatogram window). These parameters are included in the method file and might prevent integration of even correct peaks if the selected method is inappropriate. If you use the integration algorithm with the default values (Peak Width = 0.3 min., Threshold = 0.1 mv), the very narrow peaks occurring at the beginning of analysis might be ignored. Try reducing Peak Width. If this leads to detection of many small, spurious peaks, set Threshold to a value higher than the height of these spurious peaks. It must be stressed that detection of peaks always depends on an interplay of the above two parameters, and changing only one of them not always produces the expected results (see Chapter 5.2 for additional details). If none of the above two remedial measures was successful for some undetected peaks, the peaks involved are probably highly distorted so that their shape fails to satisfy the elementary conditions for peak detection. In this case use manual integration (Chromatogram - Peak - Add Positive (Negative)). Too many peaks detected For a very noisy signal the algorithm might erroneously interpret the noise as a high number of small peaks. Working with such chromatograms is very confusing and the speed of all operations decreases dramatically. Increase the value of Threshold. Make the chromatogram more lucid by reducing the number of detected peaks by appropriate setting of the integration conditions using the Rejection command. In some instances it might be advisable to ban undesirable peaks by the command Baseline - Lock. 100

101 User s Guide Resolving Problems Incorrectly detected peaks In the event the Threshold value is too high, the beginning of some peaks might be placed too high on the leading peak edge. This can be corrected by the command Chromatogram - Peak Start; alternatively, try an automatic remedy by entering a lower value of Threshold. If the Peak Width parameter is too low, a group of peaks might be separated (the peaks in the group are not separated by a vertical line). If this is the case, raise the value of Peak Width. In general, the width of detected peaks (measured as the distance between the peak beginning and end) is proportional to the value of Peak Width and inversely proportional to the value of Threshold (see Chapter Processing the Analysis for additional details) Chromatogram Modification Added peaks or some other chromatogram modification are "lost" You have probably failed to save the chromatogram after effected modifications. The integration table was mistakenly deleted or replaced by another one. The "manual integration" procedure In the event manual integration commands have been invoked (Chromatogram - Peak Add Positive (Negative)), always keep in mind that the peaks added manually, similarly to all chromatogram modifications, are stored exclusively in the integration table. How to shift the peak apex As the peak apex location is determined automatically, no instruments are available for shifting it. If it is still necessary to shift the peak apex to some other location, use the procedure described at the beginning of chapter Peak Modification on page 68. Dividing the chromatogram into regular integration areas (slices) In connection with some types of data evaluation it is necessary to divide the chromatogram into segments of equal length and integrate them separately. The simplest way how to achieve this is to forbid all existing peaks across the chromatogram by the command Baseline - Lock; then divide the chromatogram into individual segments by invoking repeatedly the command Add Positive (Negative) and "tune up" the segments 101

102 EAS32 v.1.4 Resolving Problems by editing the integration table. In some instances the commands for baseline modification may be needed. The peak beginning/end cannot be shifted to the requested location, or the required baseline modification cannot be accomplished Because of the algorithm employed for automated baseline modification a command aimed at baseline or peak modification sometimes does not conform to the user's wish. In most instances the station has detected an attempt that would result in the signal crossing the baseline and, accordingly, shifted the point in question out of the way. A new peak cannot be added You have probably attempted to add a new peak to an area occupied by the apex of another peak or to the region of tangential separation. Cancellation of a command in the integration table had unexpected consequences All your commands involving baseline or peak modification, including those that do not have the required effect, are incorporated in the integration table. At the moment you cancel a command, the originally ineffective commands may assert themselves. It is therefore recommended to maintain only functional commands in the integration table. In this manner the table will be also kept within manageable limits, a feature that will effect the sped of additional modifications Calibration Some peaks have not been calibrated New peaks can be added to an already calibrated file only by the command Add Peak but not by the command Add All. The latter calibrates all peaks from the calibration standard only for an empty calibration file; in other instances the command calibrates only the already existing peaks. A peak is erroneously calibrated as a new compound The deviation in retention time of the affected peak exceeds the size of the identification window. Increase the values stored in items Left (Right) Window or, alternately, use the reference peak method (see Chapter "Reference Peak" Method on page 86). 102

103 User s Guide Resolving Problems Note: When applying the reference peak method keep in mind that the modified retention time of a compound might differ significantly from the retention time actually measured. Thus, also if two original retention times are identical, peak identification may fail because the modified retention time may wander outside the limits given by the identification window. The calibration file is locked (no changes can be made) The calibration file involved is already open at some other channel and, accordingly, is "read-only" at this location. Neither the calibration curve nor the corresponding calibration equation is shown The item Curve Fit Type is set to Free Calibration. (The curve fit for new compounds is set by the parameter Curve Fit Type in the Defaults tab of the Calibration Options window.) 10.7 Calibrated Calculations Question marks are shown in column Weight% The value entered in parameter Weight in the chromatogram header is more than twice the sum of corrected weights in column Weight of the Result Table. The type of a peak is shown as Free in the Results table The so-called free calibration has been used. This means that the weight of the relevant compound has not been read off the calibration curve; instead, the global response factor - Resp. Factor - from the main calibration table was used. Possible reasons are the following: Free calibration has been intentionally set in item Fit Type for the affected compound. In calculation by the external standard method the equation for the requested curve fit type does not exist. The calibration curve does not pass through the origin and the calculated weight is negative. 103

104 EAS32 v.1.4 EAS32 Files and Directories 11 EAS32 Files and Directories Since the chromatographic station uses a relatively large number of directories and files of various types, and these can be moreover shared in several windows, it is advisable to present an overview Directories The EAS32 can be installed in any directory in any hard disk. After completed installation the main directory contains the following fundamental files: EAS32.EXE The executable program of the EAS32. *.DLL Dynamically loaded auxiliary libraries, containing in particular modules for data acquisition and direct chromatograph control. EAS32.HLP The main help file. *.BMP Pictures of the main station symbols. *.DTA Data files of simulated analyses used in the EAS32 demo version. EAS.PSW The file summarising the access rights of all users including their passwords. EAS.CFG The configuration file of the station. EAS.DSK A default station desktop file. *.DSK Station desktop files of individual users. *.TXT Text file containing topical information. The main directory contains the following subdirectories: COMMON Subdirectory storing common methods and report styles. PROJECTS Subdirectory containing a list of all projects. WORK1-4 The basic project directories, assigned to one station channel. Each project directory can contain the following files: *.SEQ Sample Table files. DATA Data subdirectory containing the following files: *.PRM Chromatograms CALIB Calibration subdirectory containing the following files: *.PRM Calibration standards *.CAL Calibration files A more detailed description of the structure of subdirectories is in Chapter Working with Directories and Projects on page

105 User s Guide EAS32 Files and Directories 11.2 Files Chromatograms (*.PRM) Each chromatogram contains the following: a header specifying the date and time of analysis, name of the analyst and sample parameters original raw data of the whole analysis chromatogram with the originally detected baseline method describing the progress of measurement, modifications and evaluation a reference to the name of the calibration file used summary of all chromatogram changes - Log. descriptive labels and lines Method Files (*.MET) Each method file specifies the conditions under which the chromatogram will be measured and evaluated, and contains the following: indicative information (the column and mobile phase used, the type of detector employed and its settings, etc.) parameters of the input converter, detector selection and a table for controlling the control output according to pre-defined events integration parameters integration table specifying subsequent peak and baseline modifications name of the calibration file and calculation parameters parameters used for direct chromatograph and autosampler control (optional) The contents of the method file used in the measurement is copied to each chromatogram at the moment of analysis termination Calibration Files (*.CAL) Calibration files in essence comprise a table of calibration curves. A chromatogram contains only a reference to the relevant calibration file. In other words, calibration files are not included in the chromatograms. 105

106 EAS32 v.1.4 EAS32 Files and Directories The reference to the calibration file used can be defined prior to any measurement in the current method Sample Table Files (*.SEQ) Sample Table files enable one to perform series of measurements. Each Sample Table file contains the relevant Sample Table and some auxiliary parameters. Each line of the Sample Table in fact describes the method of measurement and evaluation to be used for one or more injected samples Report Style Files (*.STY) A report style decides on what will be printed and how. The following information can be printed as part of a report: measurement description and conditions chromatograms and calibration curves tables of results Sample Tables The user can also: select page characteristics (borders, numbering) and font define the headers printed on individual pages The station is supplied with a number of pre-defined report styles. The user can modify them at will or create his own Password File (EAS.PSW) Access rights of all users are stored in this file, provided the station operates in the protected mode. To change the access rights invoke the User Accounts command from the Main window. Refer to Chapter User Accounts - Protected Mode on page 114 for additional information Configuration File (EAS.CFG) This file stores the current settings, especially the hardware configuration of the EAS32. It contains: The number of displayed channels, their names and interconnections, if any. The list and configuration of the installed A/D and D/A converters Assignment of the above devices to individual channels. 106

107 User s Guide EAS32 Files and Directories Configuration (appearance) of the Main station window. Note: A backup copy - BACKUP.CFG - is created during station installation. In the event the EAS.CFG file is damaged for any reason, you will be notified and the backup copy will be used automatically The Desktop File (EAS.DSK) The EAS.DSK file contains the following: Settings referring to size, visibility and location of all non-modal (non-dialog) windows of the station. Configuration of all graphs and tables. User-specific configuration of the station, available from the User's Options window. In the protected mode any user can use his own, dedicated DSK file. The filename is specified in item Desktop File of the User Accounts window File Sharing While working with the station the user can encounter a situation where the same file is open in several windows. This so-called file sharing can take place in the same channel or in different channels. Any change undertaken in a shared file is immediately reflected and displayed in the remaining windows where the file is currently open. A shared file can be however modified only in the channel, where it was opened first - see the next Chapter File Locking. Note: Since working files are stored in project directories, the same name of a file open in several channels does not by necessity mean that the same file is actually involved, since in most instances each channel has its own project directory. The same holds true of chromatograms in the data and calibration subdirectories File Locking To prevent collision that might corrupt the measured data, in some instances the possibility of the station working with some files is restricted. A file opened in more than one channel may be modified only in the channel where it was opened first. In the remaining channels the file is open only for reading (as 107

108 EAS32 v.1.4 EAS32 Files and Directories indicated by the inscription READ ONLY after the filename). Report style files are an exception to the above rule and can be edited simultaneously at several locations. However, only modifications effected in the window where the file was saved the last time will become permanent. The current file cannot be closed while an analysis using it is in run, nor can its parameters from the Acquisition tab of the Setup Method window be modified. While a Sample Table is in run all calibration files used are locked Marking Changes in a File Not Yet Saved For most files the notice MODIFIED after the filename indicates that the effected modifications have not been saved yet. The station always alerts to situations where such changes can be lost (e.g., when the station is closed or when a new file is being opened), and offers the option to save the changes. 108

109 User s Guide Station Settings 12 Station Settings In the interests of its most extensive utilisation the station contains a number of commands that enable customised setting, starting in the number of channels and the connected detectors, to users' access rights, to customised desktop appearance configuration, to settings of individual channels and current projects System Configuration Setting Invoke the command System - Configuration from the main window to display the system settings of the EAS32, saved in the EAS.CFG file. The base setup was already effected during the installation procedure. The command opens the System Configuration dialog box, which shows the fundamental settings in Main tab and individual settings for each channel in the corresponding tabs Channel X. Fig. 33 The Main tab of the System Configuration window How to set up the A/D converters The above Main tab of the System Configuration window displays two lists of available A/D converters and control modules (hereinafter devices): 109

110 EAS32 v.1.4 Station Settings The list to the left shows those devices whose drivers are available. Notice that this does not mean that the device itself is available! The list to the right shows the devices you want to use. Click the button or double-click the name of a device to transfer it to the righthand list - the configuration window of the device involved opens first. If an A/D or D/A converter is involved the station attempts to find the converter during the setup. The topical status is shown in the Status item. If the attempt to communicate with the converter has been successful, the device is shown in the right-hand list and its free channels can be assigned to individual channels of the station. Communication with modules for direct control of chromatographs, digital pumps and autosamplers is checked only when the channel from which the user wants to control these devices is being opened. Accordingly, during the configuration process one cannot establish whether the communication setup is correct. How to assign individual devices to the station channels? The station channel you wish to use for measuring chromatograms must have assigned the correct signal source: A channel of the A/D converter (internal INT7 or external U-PAD) Number of Channels Up to 4 channels may be set in item No. of Channels regardless of the number of channels in your version. The excess channels will be fully functional for evaluating analyses proceeding in other channels although not for data acquisition Description of Channels To make the work with the station more comprehensible the user can assign to each channel a name (in item Name) and the gas or liquid chromatography icon (in item Type of Chromatogram). Once the configuration is saved the Main station window will display the assigned names and the icon at each channel Linking Several Channels In multidetector systems it is advantageous to synchronise the measurement with several detectors. The command Link with in the tab of a channel in the System 110

111 User s Guide Station Settings Configuration window serves this purpose. Linked channels are indicated in the main station window by a grey line connecting the icons of the linked channels. The commands Run, Stop and Abort invoked in any of the linked channels will be then executed simultaneously in all linked and open channels. However, also in this instance the Snapshot command will affect only the current channel. The same holds true of external signal control User Accounts - Protected Mode The station can operate in the so-called protected mode. Advantages of the protected mode include the following: Data and the station configuration are protected against intervention by unauthorised or inexperienced persons, the station can be locked during proceeding measurement as a protection against use by unauthorised persons, several users can work with the station simultaneously and independently, customised settings can be saved for each user. 111

112 EAS32 v.1.4 Station Settings To set the protected mode, invoke the command System - User Accounts from the Main window to open the User Accounts dialog box where the changes can be effected. Fig. 34 The User Accounts Dialog Box How to establish a new user account? Open the User Accounts dialog box. Press New. Fill in the following items: Analyst Name The name of the analyst entered here will be stored in all files created by that user. Password The password to be entered by the user when opening a channel can contain up to eight characters. The preceding two items must be unique - no two users can be assigned the same name or password. Desktop File The name of the customised desktop file is entered here. The file may be shared by several users. 112

113 User s Guide Station Settings If no name is entered, the desktop file created by copying the file EAS.DSK will have the name entered in item Analyst Name. Each newly created desktop file is a copy of file EAS.DSK. What activities to ban/allow? Once a new user's account is established that user enjoys all rights. Some access rights or rights to modify specified parts of the station can be banned in the User Access Rights section. It is advisable to ban standard users from accomplishing the following activities: access to setting user passwords - User Accounts access to system configuration of the station - Configuration access to setting the station log output - Log. On the contrary, any ban on chromatogram modification - Chromatogram - is unnecessarily restrictive. How to enable working with only some channels? By means of item Access to it is possible to allow a user working with only some channels. How to restrict using or updating files by some users? The following options are available: permit only reading your files - item Read Only - or also updating - item Read/Write. If no item is checked your files will be inaccessible to other users. Under which conditions cease files to be protected? Your files are protected as long as your account exists, and cease to be protected once the mode is changed to unprotected. Files read into another EAS32 where you do not have an account will be accessible. How to cancel an existing account? In User List select the name of the user to be eliminated from the list. Invoke Remove. Select Cancel to cancel elimination from the list. All files created by the eliminated user will be then accessible to all users after the station is run the next time. If all users are eliminated the station will pass to the unprotected regime when run the next time. 113

114 EAS32 v.1.4 Station Settings How to change the access rights of a user? Select from User List the name of the user whose rights you wish to modify. All items will then refer to that user and can be changed. Cancel the effected change by the command Cancel. How to transfer the station to the unprotected mode? To do that eliminate all users from the list one by one by the command Remove. When the window is then closed the station will be unprotected and files with originally restricted access will be accessible. What if you forget the password? In this instance transfer the station to the unprotected mode by copying the file EAS.PSW from the PGM directory on the installation disk to the main station directory. This remedy however cancels all user access rights previously set Channel Locking In the protected mode any open channel can be locked to protect it against unauthorised intervention during the user's absence. To lock a channel invoke the Lock command from the Lock menu in the Main window. All activities continue unabated in the locked channel (running a Sample Table, printing a report, batch processing, etc.). Unlock the channel by either repeatedly invoking the Lock command or by clicking at the chromatograph symbol and entering the password of the user who opened that channel User Settings The station automatically saves the settings of visibility, size and location of all nonmodal windows, the configuration of all graphs and tables, and all user-specific settings available from the User's Options window. The settings are saved in the desktop file EAS.DSK. In the protected mode the settings are saved in the file specified in item Desktop File of each user listed in the User Accounts window. Thus, in the protected regime each user can have a customised setting of the station appearance. The settings of all windows, graphs and tables are channel-specific. The parameters specified in the User's Options are common to all channels used by a user. 114

115 User s Guide Station Settings 12.5 Working with Directories and Projects The station enables a base directory to be assigned to each channel as well as storing analyses in different projects Channel Directory Selection Invoke the command Directories in the Main station window to specify for each channel a different base directory for storing projects. The above command is especially suitable if the EAS32 is a part of a computer network where either several stations or several evaluation programs EAS32 Evaluation are operative. By default all channels use the main station directory, usually C:\EAS32. Channel directories are stored in the configuration file EAS.CFG. At least one project directory is assumed to exist in the channel directory; the former is described in subdirectory PROJECT. If no channel directory, no PROJECT subdirectory and no project directory exists you will be queried whether you wish to create the corresponding directory or project Projects A project is in effect a directory earmarked for storing analyses and auxiliary files. Sample table files and reports styles are stored directly in the project directory. The data subdirectory stores chromatograms, the calibration subdirectory stores calibration standards and calibration files. To switch between the calibration and data subdirectories when selecting files click either of icons and in the File Open dialog box. By default, files from the current project are offered in each channel. Use the icons and commands from the File Open dialog box to select a file from any other directory. To facilitate sharing files common to several projects (e.g. report styles) the station contains the COMMON directory. Click icon iii in the File Open dialog box to switch to the COMMON directory. How to establish a new project? Create a new project in the Channel window using the command File Project, which opens the window Project Setup, and in this window press the button New. Having first entered the project name you may modify the names of the data and calibration subdirectories. 115

116 EAS32 v.1.4 Station Settings How to store chromatograms and calibration files in the same directory? Select the same name for the two subdirectories in the Project Setup dialog box (items Analysis and Calibration Subdir). How to open another project? Select a new project by the command File - Project - Open. A project can be opened in several channels and can be shared by other EAS32 programs across a computer network Backing up and Restoring Files and Projects The EAS32 provides for backing up and restoring all data and working files Backup The File - Archive command from the Channel window is earmarked for backing up data. When invoked the command opens the Archive tab of the Backup dialog box. Fig. 35 The Backup - Archive Dialog Box 116

117 User s Guide Station Settings How to select the type of files to be archived? The type of files to be archived is specified by item File Type. In addition to all EAS32 files, entire projects can be also archived, i.e., all files identified in a specified project directory and all its subdirectories. How to select files to be archived? The list shown in the upper part of the Archive tab contains all files of the selected type from the current project. Select a file by clicking the filename. Several files can be selected by clicking their names one by one while holding down Ctrl. The meaning of the Select All Files button is self-evident. Where will be the files archived? Specify the destination (another directory, disk or computer) in item Target. The path can be either entered directly or selected by the button. In what format are files stored in the target location? If item Without Compressing is checked, files will be stored unchanged, as individual files, otherwise all archived files will be compressed and stored as a single file under a selected name. Will the files be copied or moved? If item Moved to Archive is checked, the file(s) will be moved to the destination (i.e., deleted from the original directory). What is the total size of files to be archived? The overall size is indicated above the list of the selected files, in item Selected Size. How to sort files in the list? Double-click the header of each column to sort the entire table according to that column. Double-click again to sort the files in the reverse order. Backing up calibration standards If chromatograms are to be archived, the Calibration Standard item displays the contents of the calibration subdirectory. Archiving files from the COMMON directory Check item From Common to archive files stored in the COMMON directory. 117

118 EAS32 v.1.4 Station Settings How to delete files? Invoke the command Delete Selected Files. This command cannot be reversed from within the EAS32! Restoring Files The command File Restore from the channel window restores files from any directory to the current project. When invoked the Restore tab from the Backup dialog box opens; it is very similar to the Archive tab. Fig. 36 The Backup - Restore dialog box. To which location will be files restored? The Restore command always restores files to the current project whose name is shown in the title bar of the Backup dialog box. From which location will be files restored? The source location (other directory, disk, or computer) can be specified in item Source. Enter the path either directly or use the button to browse. The other items and commands are analogous to those listed in the Archive tab described in Chapter Backing up and Restoring Files and Projects on page