Screening for 926 Pesticides and Endocrine Disruptors by GC/MS with Deconvolution Reporting Software and a New Pesticide Library Application Note

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Screening for 926 Pesticides and Endocrine Disruptors by GC/MS with Deconvolution Reporting Software and a New Pesticide Library Application Note Screening for 926 Pesticides and Endocrine Disruptors by GC/MS with Deconvolution Reporting Software and a New Pesticide Library Application Note Food and Environmental Authors endocrine disrupters in about two minutes per sample. Deconvolution helps identify pesticides that are buried in the chromatogram by co-extracted materials. The new Philip L. Wylie database was compared to the smaller one for the DRS Agilent Technologies, Inc. analysis of 17 surface water samples. With the new data- base, DRS found 99 pesticides, metabolites, fire retar- 2850 Centerville Rd. dants, and related contaminants that were not contained Wilmington, DE 19808-1610 in the original RTL Pesticide and Endocrine Disruptor USA Library. Abstract Introduction An updated and greatly expanded collection of mass Several years ago Agilent Technologies introduced spectral libraries has been introduced, replacing Agilent’s Retention Time Locking (RTL) for gas chromatog- RTL Pesticide Library and DRS pesticide solution. The raphy (GC) and GC with mass spectral detection new library contains 926 pesticides, endocrine disruptors, (GC/MS). RTL software makes it possible to repro- and related compounds – 359 more than the original duce retention times from run-to-run on any library. Included are all compounds specified for GC/MS Agilent GC or GC/MS, in any laboratory in the analysis in the new Japanese “Positive List” regulations. world, so long as the same nominal method and GC All compounds have locked retention times that can be column are used (1). Since any laboratory can accurately reproduced using an Agilent GC/MS system with the ChemStation's Retention Time Locking software. reproduce retention times generated in another, it The new Database can be used as a standard GC/MS is possible to create mass spectral libraries that library for compound identification or with Agilent's contain locked retention times. By locking their Screener software for identifications based upon reten- method to the published database, users can tion time and mass spectral matching. The greatest bene- screen GC/MS files for all of the library’s com- fit accrues when these libraries are used with Agilent’s pounds. “Hits” are required to have the correct new version of Deconvolution Reporting Software (part retention time as well as the correct spectrum, number G1716AA version A.03.00). This solution allows which eliminates many false positives and gives one to screen GC/MS files for all 926 pesticides and more confidence in compound identifications (2). More recently, Agilent introduced Deconvolution • A screener database for use with Agilent’s pow- Reporting Software (DRS) that incorporates mass erful screener software that is integrated into spectral deconvolution with conventional library the GC/MS ChemStation searching and quantification. DRS results from a • Locked Retention Times for all 926 compounds marriage of three different GC/MS software that any Agilent 5975 or 5973 GC/MS user can packages: reproduce in their laboratory 1) The Agilent GC/MS ChemStation, • Files for use with Agilent’s G1716AA (A.03.00) 2) The National Institute of Standards and Tech- Deconvolution Reporting Software nology (NIST) Mass Spectral Search Program • An e-method that can be loaded into Agilent’s with the NIST ‘05 MS Library, and G1701DA (version D.02.00 SP1 or higher) with 3) The Automated Mass Spectral Deconvolution instrument parameters for acquiring GC/MS and Identification System (AMDIS) software, files and analyzing the data with DRS. These also from NIST. parameters are listed in Table 1. The original DRS software was intended to be a • Example files comprehensive solution for pesticide analysis and, • Application notes therefore, included the mass spectra (in AMDIS format) and locked retention times for 567 pesti- On November 29, 2005, the Japanese Government cides and suspected endocrine disrupters (3). published a “Positive List” system for the regula- tion of pesticides, feed additives, and veterinary Recently, Agilent introduced an updated and drugs. Maximum Residue Limits (MRL) have been greatly expanded Pesticide and Endocrine Disrup- set for 758 chemicals while 65 others have been tor Database (part number G1672AA) that now exempted from regulation. Fifteen substances must contains 926 entries. This represents the addition have no detectable residues. Other agricultural of 359 new compounds to the original library. At chemicals not mentioned have a uniform MRL of the same time, Agilent introduced a new version of 0.01 ppm (4). This new regulation is scheduled to the DRS software (part number G1716AA version take effect on May 29, 2006. A.03.00) that can be used with any Agilent-provided or user-developed DRS library. Of the pesticides in the Japanese Positive List, 265 are to be analyzed by GC/MS. The new G1672AA Pesticide and Endocrine Disruptor Database Contents Pesticide library contains mass spectra and locked retention times for all of these compounds. Thus, a The G1672AA Pesticide and Endocrine Disruptor laboratory could screen for all 265 “positive list” Database contains virtually all GC-able pesticides, compounds and several hundred more pesticides including those introduced very recently. In addi- in just 1–3 minutes after the GC/MS run. tion, the database includes numerous metabolites, more endocrine disruptors, important PCBs and PAHs, certain dyes (for example, Sudan Red), Experimental synthetic musk compounds, and several Table 1 lists the instrumentation, software, and organophosphorus fire retardants. analytical parameters used by Agilent for pesticide This new database includes: analysis. Depending upon the desired injection volume, a PTV inlet or split/splitless inlet can be • A conventional mass spectral library for use used. with Agilent GC/MS ChemStations 2 Table 1. Instrumentation and Conditions of Analysis Gas Chromatograph Agilent 6890N Automatic Sampler Agilent 7683 Injector and AutoSampler Inlet Agilent PTV operated in the solvent vent mode or Split/Splitless Column Agilent 30 m × 0.25 mm × 0.25 µm HP-5MSi (part number 19091S-433i) Carrier gas Helium in the constant pressure mode Retention time locking Chlorpyrifos-methyl locked to 16.596 min (nominal column head pressure = 17.1 psi) Oven temperature program 70 °C (2 min), 25 °C/min to 150 °C (0 min), 3 °C /min to 200 °C (0 min), 8 °C /min to 280 °C (10–15 min) PTV inlet parameters Temp program: 40 °C (0.25 min), 1600 °C/min to 250 °C (2 min); Vent time: 0.2 min; Vent flow: 200 mL/min; Vent pressure: 0.0 psi; Purge flow: 60.0 mL/min; Purge time: 2.00 min Injection volume 15 µL (using a 50-µL syringe) Mass Selective Detector Agilent 5975 inert Tune file Atune.u Mode Scan (or SIM with SIM DRS library) Scan range 50–550 u Source, quad, transfer line 230, 150, and 280 °C, respectively temperatures Solvent delay 4.00 min Multiplier voltage Autotune voltage Software GC/MSD ChemStation Agilent part number G1701DA (version D02.00 sp1 or higher) Deconvolution Reporting Software Agilent part number G1716AA (version A.03.00) Deconvolution Reporting Software Library searching software NIST MS Search (version 2.0d or greater) (comes with NIST '05 mass spectral library – Agilent part number G1033A) Deconvolution software Automated Mass Spectral Deconvolution and Identification Software (AMDIS_32 version 2.62 or greater; comes with NIST '05 mass spectral library – Agilent part number G1033A) MS Libraries NIST ‘05 mass spectral library (Agilent part number G1033A) Agilent RTL Pesticide and Endocrine Disruptor Libraries in Agilent and NIST formats (part number G1672AA) Results and Discussion that is supplied with the DRS software. The DRS then sends the data file to AMDIS, which deconvo- DRS, which has been described in preceding lutes the spectra and searches the Agilent RTL papers (3,5,6), can be summarized as follows: Pesticide Library using the deconvoluted full spec- tra. A filter can be set in AMDIS, which requires Three separate, but complimentary, data analysis the analyte’s retention time to fall within a user- steps are combined into the DRS. First, the GC/MS specified time window. Because RTL is used to ChemStation software performs a normal quanti- reproduce the RTL database retention times with tative analysis for target pesticides using a target high precision, this window can be quite small – ion and up to three qualifiers. An amount is typically 10–20 seconds. Finally, the deconvoluted reported for all calibrated compounds that are spectra for all of the targets found by AMDIS are detected. For other compounds in the database, an searched against the 147,000-compound NIST mass estimate of their concentration can be reported spectral library for confirmation; for this step, based upon an average pesticide response factor there is no retention time requirement. 3 This approach was rapidly adopted by many labo- Advantages of Deconvolution ratories because of its ability to identify pesticides in complex chromatograms containing high levels Figure 1 shows a screen from AMDIS that illus- of co-extracted interferences. Indeed, the solution trates the power of this deconvolution software. proved to be so useful that users began to create The white trace in Figure 1A is the total ion chro- their own DRS libraries (7). Therefore, the DRS matogram while the other three are extracted ions was unbundled from the pesticide database so that of a deconvoluted peak (a “component” in AMDIS it could be used with any agilent-provided or terminology). Note that the TIC and extracted ions user-created database. are not scaled to each other and this component is actually obscured by co-eluting compounds. Figure The original 567-compound RTL Pesticide Library 1B juxtaposes the deconvoluted component spec- (G1049A) included pesticides, a few metabolites, trum (white) with the complete “undeconvoluted” and most of the GC-amenable endocrine disruptors spectrum (black). Clearly, this component is buried that were known at the time. The new version of under co-eluting peaks that would ordinarily the library includes many more pesticides, obscure the analyte.
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