Retention Time Locking Operation

Retention Time Locking Operation Retention Time Locking Database 32-1 What is Retention Time Locking? The ability to precisely match chromatographic retention times in any Agilent GC system to those in another Agilent GC system with the same nominal capillary column 32-2

What is Retention Time Locking Database? RTL Database allows the user to identify unknowns in a sample without standard Any given analyte will always elute at the same One method can be used for multiple sample types All GC system configurations can make use the same database Databases can be easily ported to other columns with GC method translation 32-3 When Should We Need RTL Databases? RTL Databases can be used to screen for unknown analyte without distortion of peak elution time Preliminary analyte identification is possible without standards 32-4

What is the Agilent 567 Pesticide Library? A mass spectral database of 567 Pesticides and Endocrine disrupters generated on 5973(or 5975) system under RTL conditions, which includes retention times and mass spectral information upon RTL screening database (RTLPEST.SCD) Retention time database of 567 Pesticides and Endocrine disrupters generated on Agilent 6850, 6890 system under RTL conditions, which includes retention times and containing elements upon RT Table (RTGCPST.RTT) 32-5 Small Portion of the Agilent 567 Pestide Library Contains 567 compounds 32-6

What is the Agilent 926 Pesticide Library? More updated and greatly expanded Pesticide and Endocrine Disruptor Database/Library, which contains 926 pesticides, metabolites, and endocrine disruptors, along with other food/environmental contaminants With Harzardous Chemical Compounds(731 entries), These new libraries can be used with the Agilent G1701DA GC/MS ChemStation as stand-alone mass spectral databases. But, their true power is revealed when combined with the G1716AA DRS software. 32-7 Overview RTL Database Searching 1. Locking GC RT with pesticide RT table by RTL The necessary steps for locking a method described in How does RTL work and Method Translation 2. Run GC chromatograms and GC database search 3. Hit possible compounds result in unknown pesticide identified Cf. To get more reliable results with spectrum, GC/MSD is used 32-8

Installation of RTL Databases [NOTE] Before starting 1. make sure that the Retention Time Locking software is installed in the ChemStation. Complete the installation and become familiar with the RTL software before proceeding. 2. Copy the file; HPGCPST.RTT and RT_SRCH.MCX on the RTL Pesticide Library CD (G2081AA) to C:\HPCHEM\RTL directory 32-9 Universal Pesticide Method for GC, GC/MSD For HP-5MS columns 30m x 0.25mm x 0.25μm Flexible injection schemes: (Retention gap-)coc, (LVI-)PTV, Split/Splitless Pursed Splitless 280 C (10 min) Any detector: MSD, ECD, NPD, FPD 200 C (0 min) 8 C/min 150 C (0 min) 3 C/min 70 C (2 min) 25 C/min Total run time = 41.87 min 32-10

Data Analysis 32-11 Click on plot On real data file 32-12

Double Click 32-13 1.Load Table - Choose RT Table Searching RT Database - Search RT and Search Window are auto-filled 32-14

Load RT Table 32-15 Actual Pesticide Database Search - Using an unknown s RT only yields 6 possibilities 32-16

2.Elements in Peak Set Filters - Contain or not contain element - Detector 3.Detectors that detect peak [NOTICE] This entry only appears if the method uses an element-selective detector 32-17 Searching RT Database - Using RT and Elemental Qualifiers Unknown has Cl,N,O,P,S Unknown does not have Br,F 32-18

Actual Pesticide Database Search - Using unknown s RT and element information yield just 1 possibility Qualifier : 16.582 min peak has Cl,N,O,P,S But does not have Br,F Peak is Chloropyriphos-methyl: Actual RT = 16.582 Database RT = 16.596 32-19 Actual Pesticide Database Search - Another Peak 32-20

32-21 Actual Pesticide Database Search - Using an unknown s RT only yields just 1 possibility Qualifier : No Restriction Peak is Piperalin: Actual RT = 27.264 Database RT = 27.250 32-22

Result report of RT Search on Peak at 27.264 minutes 32-23 Type in time On real data file - Another RTSearch option 32-24

1.Peaks RT 2.Search window Searching RT Database - You must entered Search RT and Search Window, minute 32-25 Use Table for Rapid Identification Scaled and locked methods make analyses easier Master RT Table 1.263 methanol 1.323 acetone 1.463 ethanol 2.263 darnitol 3.263 goodbyeall 1.263 methanol 1.323 acetone 1.463 ethanol 2.263 darnitol 3.263 goodbyeall GC1 GC2 GC3 GC4 FID FPD AED MSD 32-26

RTL Databases (different systems) 6890 Systems Inlet Detector Injection Technique Dichlorvos Chlorpyrifos Mirex methyl PTV MSD Splitless 5.897 16.593 29.800 PTV µ-ecd Split 5.798 16.576 29.876 S/SL FID Splitless 5.797 16.587 29.856 S/SL AED Splitless 5.829 16.600 29.839 PTV µ-ecd Split 5.860 16.597 29.864 PTV µ-ecd Cold Splitless 5.862 16.589 29.867 S/SL Dual FPD Splitless 5.814 16.596 NA S/SL AED Splitless 5.837 16.604 29.851 COC MSD On-Column 5.862 16.607 29.836 S/SL NPD Splitless 5.814 16.596 NA HI - Lo 0.100 0.028 0.076 Average 5.837 16.595 29.849 STDEV 0.033 0.009 0.024 Target = Chlorpyrifos-Methyl @ 16.596 32-27 Use RTL Databases Worldwide GC1 GC2 GC3 GC4 GC5 GC6 Lab 1, USA GC1 GC2 GC3 Master RTL Database 1.263 methanol 1.323 acetone 1.463 ethanol 2.263 darnitol 3.263 goodbyeall 1.263 methanol 1.323 acetone 1.463 ethanol 2.263 darnitol 3.263 goodbyeall GC1 GC2 GC3 GC4 GC5 GC6 Lab 1, UK GC1 GC2 GC3 GC4 GC5 GC6 Lab 1, JPN GC4 GC5 GC6 Lab 1, FRA 32-28

Some Users of the Agilent Universal Pesticide Method Florida Department of Food and Agriculture Quest International (foods and flavors) Ocean Spray (juices) Gerber (baby foods) California Department of Food and Agriculture Oregon Department of Agriculture United States Bureau of Alcohol, Tobacco, and Firearms (ATF) (Pesticides in imported wine) 32-29 Which RTL Databases are Available Agilent Order Possible Pesticide and Endocrine Disrupters (G1049A for GC/MSD, G2081AA for GC, 567 entries) Expended Pesticide and Endocrine Disrupters(G1672AA, 926 entries) Hazardous Chemical Compounds(G1671AA, 731 entries) The following databases are FREE from the Agilent Website PCB Congeners (209 entries) Volatiles (66 entries) Phenols (70 entries) Forensic and Toxicology (277 entries) 32-30

RTL Databases Benefits Enables construction of universal retention time databases Rapid identification and confirmation of unknowns Improved analytical results between labs and over time Lower cost per sample Improved throughtput Simplified data interpretation Scalable to fast GC 32-31 Thank you very much for your attention! 32-32