Exceed the Requirements of Standard Environmental Methods using a High Performance GC-TOFMS. Jonathan D. Byer
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1 Exceed the Requirements of Standard Environmental Methods using a High Performance GC-TOFMS Jonathan D. Byer
2 Outline Pesticides in Fruits and Vegetables SANTE/11945/215 US EPA Method 827 Semi-volatile organic compounds by GC/MS US EPA Method Organic compounds in drinking water, source water, or water in any stage of drinking water treatment by GC/MS
3 Pesticides in Fruits and Vegetables
4 SANTE/11945/215 MS Requirements
5 SANTE/11945/215 MS Requirements
6 LECO Pegasus BT A better way: using the Pegasus BT GC-TOFMS
7 Data Processing Methods Non-Target Deconvolution: Quantitative and/or Qualitative 1.8e8.96e8.84e8.72e8.6e8.48e8.36e8.24e8.12e8 Time (min) TIC Target Analyte Find: Quantitative and/or Screening.96e7.84e7.72e7.6e7.48e7.36e7.24e7.12e7 Time (min) AIC Specify R.T. Window to Peak Find, Minimum S/N, Minimum number of ions in mass spectrum, Signal ± Mass Tolerance, Library Search Create a target list Specify Analyte Name, R.T. Window, and Mass(es) ± Mass Tolerance
8 Data Processing Methods Non-Target Deconvolution: Quantitative and/or Qualitative 1.8e8.96e8.84e8.72e8.6e8.48e8.36e8.24e8.12e8 Time (min) TIC Target Analyte Find: Quantitative and/or Screening.96e7.84e7.72e7.6e7.48e7.36e7.24e7.12e7 Time (min) AIC Peak True - sample "MR Pest5 2 SR 1:3", Etridiazole, at min, Height (Counts) Library Hit - Similarity: 94 - Library: replib - Etridiazole, Abundance M/Z
9 Data Processing Methods Non-Target Deconvolution: Quantitative and/or Qualitative 1.8e8.96e8.84e8.72e8.6e8.48e8.36e8.24e8.12e8 Time (min) TIC Peak True - sample "MR Pest5 2 SR 1:3", Etridiazole, at min, Height (Counts) Target Analyte Find: Quantitative and/or Screening.96e7.84e7.72e7.6e7.48e7.36e7.24e7.12e7 Time (min) AIC Peak True - sample "MR Pest5 2 SR 1:2", Etridiazole, at min, Area (Counts) 1 5 M/Z Library Hit - Similarity: 94 - Library: replib - Etridiazole, Abundance e5 6e5 5e5 4e5 3e5 2e5 1e5 M/Z
10 Data Processing Methods Non-Target Deconvolution: Quantitative and/or Qualitative Target Analyte Find: Quantitative and/or Screening 1.8e8.96e7.96e8.84e7.84e8 +.72e7.72e8.6e7.6e8.48e7.48e8.36e7.36e8.24e7.24e8.12e7.12e8 TIC AIC Time (min) Time (min) Peak True - sample "MR Pest5 2 SR 1:3", Etridiazole, at min, Height (Counts) Peak True - sample "MR Pest5 2 SR 1:2", Etridiazole, at min, Area (Counts) 1 5 M/Z Library Hit - Similarity: 94 - Library: replib - Etridiazole, Abundance e5 6e5 5e5 4e5 3e5 2e5 1e5 M/Z
11 Data Processing Options: Quantitative Target List
12 MS Criteria for SANTE/11945/215 To meet method requirements, must pass: 1. Minimum of 3 ions 2. S/N > 3 3. Ions XICs must overlap 4. ±3% ion ratio across calibration range Blueberry Gas Chromatograph Injection Carrier Gas Column LECO L-PAL3 Autosampler with Agilent MMI, 789B GC 1 µl Cold Splitless, 5 C to C/min 2. ml/min, Constant Flow Rxi-5MS, 3 m x.25 mm i.d. x.25 µm coating Spinach Oven Program 7 C (1 min), to 15 5 C/min, to 33 1 C/min (2 min) Transfer Line 3 C Mass Spectrometer LECO Pegasus BT Ion Source Temperature 2 C Mass Range m/z Acquisition Rate 2 spectra/s
13 MS Criteria for SANTE/11945/215 Ion ratio <±3% across calibration range.1 ng/g.5 ng/g 1. ng/g 5. ng/g 1 ng/g 5 ng/g 1 ng/g 5 ng/g Hexachlorobenzene
14 MS Criteria for SANTE/11945/215 Ion ratio <±3% across calibration range.1 ng/g.5 ng/g 1. ng/g 5. ng/g 1 ng/g 5 ng/g 1 ng/g 5 ng/g Hexachlorobenzene
15 MS Criteria for SANTE/11945/215 Ion ratio <±3% across calibration range.1 ng/g.5 ng/g 1. ng/g 5. ng/g 1 ng/g 5 ng/g 1 ng/g 5 ng/g Hexachlorobenzene
16 MS Criteria for SANTE/11945/215 More than 3 ions, Ions overlap in XIC, and S/N > 3.5 ng/g.5 ng/g Hexachlorobenzene
17 MS Criteria for SANTE/11945/215 Hexachlorobenzene Endosulfan sulfate Folpet.5 to 5 ng/g.5 to 5 ng/g 5. to 5 ng/g r =.9968 r =.995 r =.9987
18 MS Criteria for SANTE/11945/215 Hexachlorobenzene Endosulfan sulfate Folpet.5 to 5 ng/g 1. to 5 ng/g 5. to 5 ng/g r =.9998 r =.9971 r =.9947
19 Detection of Pesticides in Strawberry LODs in Strawberry in ng/g Target Analyte Find: Quantitative and/or Screening.96e7.84e7.72e7.6e7.48e7.36e7.24e7.12e7 Time (min) AIC Pyriproxyfen Iprodione Propargite Tebuconazole Hexazinone Fenarimol Lenacil Fluridone Etofenprox Etridiazole Atrazine Terbuthylazine Terbacil Vinclozoline MGK 264 Penconazole Fipronil Procymidone Paclobutrazol Bupirimate Flusilazole Myclobutanil Fludioxonil Tricyclazole Flutriafol * Incurred pesticides were removed from the figure
20 TIC Superior Selectivity in Matrix
21 Superior Selectivity in Matrix TIC XIC (159.±1)
22 Superior Selectivity in Matrix TIC XIC (159.±.3) XIC (159.±1)
23 Superior Selectivity in Matrix TIC XIC (159.±.3) XIC (159.±1) XIC (158.97±.95)
24 Superior Selectivity in Matrix TIC XIC (159.±.3) XIC (159.±1) XIC (158.97±.95) ±6 ppm
25 912/1 Leverage Profile Data
26 Leverage Profile Data
27 Leverage Profile Data ±.95 Da 6 ppm
28 Leverage Profile Data
29 Leverage Profile Data
30 Leverage Profile Data ±.17 Da 6 ppm
31 Superior Selectivity in Matrix TIC XIC (159.±.3) XIC (159.±1) XIC (158.97±.95) ±6 ppm
32 Data Processing Options: Qualitative + Quantitative for Conc. > low ppb Non-Target Deconvolution + Library Database Searching
33 Time (min) Automatic Peak Find: NTD 23 Pesticides in 8 mins 3.6e6 Additional Tools: Deconvolution Eight pesticides identified within 5 sec R.T. 3.e6 2.4e6 Lindane Terbuthylazine Terbufos Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos 15 m x.25 mmid x.25 µm df Rxi-5MS (Restek) 1.8e6 2.4e7 2.1e7 TIC 1.2e6.6e6 XIC 1.8e7 1.5e7 1.2e7.9e7 Time (min) Additional Tools: Deconvolution Eight pesticides identified within 5 sec R.T..6e7.3e7
34 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 1.2e6.6e6 Time (min) Lindane * Terbuthylazine Terbufos 15 m x.25 mmid x.25 µm df Rxi-5MS Profluralin XIC Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", ë-lindane, at min, Area ( Abundance) Cl Cl Cl Cl Cl 7 Cl 6 5 Lindane M/Z Library Hit - Similarity: Library: replib - ë-lindane, Abundance 896/
35 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 Lindane Terbuthylazine Terbufos Eight identified pesticides in 5 seconds Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Terbuthylazine, at min, Ar ea (Abundance) NH N NH N N Cl Terbuthylazine 1.8e6 1.2e6.6e6 Time (min) * XIC Library Hit - Similarity: Library: replib - Terbuthylazine, Abundance / M/Z
36 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 1.2e6.6e6 Time (min) Lindane Terbuthylazine Terbufos * Eight identified pesticides in 57.7 seconds XIC Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Terbufos, at min, Area ( Abundance) M/Z Library Hit - Similarity: Library: replib - Terbufos, Abundance 872/ S S S P O O Terbufos
37 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 1.2e6.6e6 Time (min) Lindane Terbuthylazine Terbufos Eight identified pesticides Abundance) in 5 seconds XIC Profluralin * Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Profluralin, at min, Area ( M/Z Library Hit - Similarity: Library: mainlib - Profluralin, Abundance /1 O O N F F F N O NO Profluralin
38 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 Lindane Terbuthylazine Terbufos Eight identified pesticides in 5 seconds Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Benzene, pentachloronitro-, at min, Area (Abundance) Library Hit - Similarity: Library: replib - Benzene, pentachloronitro-, Abundance Pentachloronitrobenzene Cl Cl O Cl N O Cl Cl 1.2e6.6e6 Time (min) XIC * M/Z 3 852/
39 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 Lindane Terbuthylazine Terbufos Eight identified pesticides ea (Abundance) in 5 seconds Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Propyzamide, at min, Ar Library Hit - Similarity: Library: replib - Propyzamide, Abundance Propyzamide Cl Cl O NH 1.2e6.6e6 Time (min) XIC * M/Z 823/
40 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 1.2e6.6e6 Time (min) Lindane Terbuthylazine Terbufos Eight identified pesticides 381 min, Area in 5 (Abundance) seconds Pentachlorobenzonitrile XIC Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- * Fonofos Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Benzonitrile, pentachloro-, at M/Z Cl Cl Library Hit - Similarity: Library: replib - Benzonitrile, pentachloro-, Abundance 862/1 N Cl Cl Cl
41 Automatic Peak Find: NTD 3.6e6 3.e6 2.4e6 1.8e6 1.2e6.6e6 Time (min) Lindane Terbuthylazine Terbufos Eight identified pesticides bundance) in 5 seconds XIC Profluralin Benzene, pentachloronitro- Propyzamide Benzonitrile, pentachloro- Fonofos * Peak True - sample "MR All 1 PSL OT7 C, 2 psi", Fonofos, at min, Area (A M/Z Library Hit - Similarity: 89 - Library: mainlib - Fonofos, Abundance Fonofos 89/1 S S P O
42 Non-Target Incurred Pesticide Peak True - sample "Blank - BlueBerry_2", BOSCALID, at min, Height (Counts) 3.6e6 2.4e6 1.2e6 3.6e6 2.4e6 1.2e6 3.6e6 2.4e6 1.2e6 Time (min) XIC(14.2±.8) Blank - Tomato_2 XIC(14.2±.8) Blank - Spinach_ XIC(14.2±.8) Blank - BlueBerry_2 1.6e6 1.4e6 1.2e6 1.e6.8e6.6e6.4e6.2e Library Hit - Similarity: Library: mainlib - BOSCALID, Abundance 14 O NH Cl N Cl 868/1 M/Z
43 Pesticide Summary 2.4e8 2.1e8 Time (min) Propoxur (Incurred) XIC(11.4±.7) Increase productivity Quantitate target analytes at low concentration Overcome matrix challenges Chromatography High quality MS Data Effective processing (e.g., Deconvolution) Mass Selectivity 1.8e8 1.5e8 1.2e8.9e8.6e8 12 replicate injections of a cabbage sample.3e8 Time (min) TIC
44 USEPA Method 827
45 USEPA Method 827 Semivolatile Organic Compounds by GC/MS Gas Chromatograph LECO L-PAL3 Autosampler with Agilent 789B GC Injection 1 µl injection, Split 27 C Carrier Gas 1.4 ml/min, Constant Flow Column ZB-SemiVolatiles, 3 m x.25 mm i.d. x.25 µm coating (Phenomenex, Torrance, CA, USA) Oven Program 7 C (1 min), to C/min, to 35 3 C/min, to 32 3 C/min (1 min) Transfer Line 3 C Mass Spectrometer LECO Pegasus BT Ion Source Temperature 25 C Mass Range 3-65 m/z Acquisition Rate 1 spectra/s Total ion chromatogram (TIC) of a 5 ppm standard containing 9 components
46 USEPA Method 827 To meet method requirements, must pass: 1. DFTPP Tuning 2. GC/MS Check Solution 3. Linearity of Calibration 4. Calibration Verification 5. QC Check Standard Gas Chromatograph LECO L-PAL3 Autosampler with Agilent 789B GC Injection 1 µl injection, Split 27 C Carrier Gas 1.4 ml/min, Constant Flow Column ZB-SemiVolatiles, 3 m x.25 mm i.d. x.25 µm coating (Phenomenex, Torrance, CA, USA) Oven Program 7 C (1 min), to C/min, to 35 3 C/min, to 32 3 C/min (1 min) Transfer Line 3 C Mass Spectrometer LECO Pegasus BT Ion Source Temperature 25 C Mass Range 3-65 m/z Acquisition Rate 1 spectra/s
47 USEPA Method Decafluorotriphenylphosphine Caliper - sample "GCMS Tune Mix 5 ng_ul Split 2", min min, Height (Abundance) 1.5e7 1.2e7.9e7.6e7.3e7 Time (min) XIC(442.±.27) M/Z Library Hit - Similarity: 97 - Library: WR1 - Phosphine, bis(pentafluorophenyl)phenyl-, Abundance
48 USEPA Method Decafluorotriphenylphosphine Average of 1 replicate injections of 5 ng/µl, Split 2:1 1.5e7 1.2e7.9e7.6e7.3e7 Time (min) XIC(442.±.27) Mass Ion Abundance Criteria Mean STDEV Min Max Criteria % of Base Peak PASS 68 < 2% of mass PASS 7 < 2% of mass PASS % of Base Peak PASS 197 < 2% of mass PASS 198 Base Peak, or > 5% of Mass 442 Base Peak PASS % of mass PASS % of Base Peak PASS 365 > 1% of mass PASS 441 Present but < 24% of mass PASS 442 Base Peak, or > 5% of mass PASS % of mass PASS
49 USEPA Method GC/MS Check Solution 1 replicate injections of 5 ng/µl, Split 2:1 Tailing Factor < 2 % DDT Breakdown < 1%
50 USEPA Method 827: Initial Calibration
51 USEPA Method Linearity of Target Analytes % RSD RF % RSD RF must be < 2 % 84 Compounds 91% of Compounds had % RSD RF < 1% 1 5 Compounds Concentration range from.5 to 5 ppm, 1 point curve in replicate
52 USEPA Method Linearity of Target Analytes 35 3 % RSD RF Compounds Concentration range from.5 to 5 ppm, 1 point curve in replicate
53 USEPA Method Linearity of Target Analytes % RSD RF Ratio y= x r = Benz[a]anthracene r > ppm 827 Split 2_ ppm 827 Split 2_6 2 ppm 827 Split 2 2 ppm 827 Split 2 Compounds Concentration Concentration range from.5 to 5 ppm, 1 point curve in replicate
54 USEPA Method Calibration Verification 2 1 % D % Difference < ± 2 % Concentration (ppm) Concentration (ppm) 2,4-Dimethylphenol
55 USEPA Method Check Standard 84 Compounds % D 1 % Difference < ± 2 % Compound 827 Matrix Spike Mix Calculated Using Calibration Curve
56 Summary: USEPA Method 827 To meet method requirements, must pass: 1. DFTPP Tuning 2. GC/MS Check Solution 3. Linearity of Calibration 4. Calibration Verification 5. QC Check Standard
57 USEPA Method
58 USEPA Method e9 2.1e9 1.8e9 1.5e9 1.2e9.9e9.6e9.3e9 Time (min) Fortified Water Extract 258 Analytes Gas Chromatograph TIC LECO L-PAL3 Autosampler with Agilent 789B GC Injection 1 µl cold splitless injection, 5 C (.1 min) to C Carrier Gas 2. ml/min, Constant Flow Column Rxi-5MS, 3 m x.25 mm i.d. x.25 µm coating (Bellefonte, PA, USA) Oven Program 7 C (1 min), to 15 5 C/min, to 33 1 C/min (2 min) Transfer Line 3 C Mass Spectrometer LECO Pegasus BT Ion Source Temperature 28 C Mass Range m/z Acquisition Rate 8 spectra/s
59 USEPA Method LODs in Water Extract Compounds <.5 ppb 2 ppb 5 ppb 5 1 ppb 25 ppb On Column [Conc.] in Water Extract Statistically derived LOD calculated from five replicate injections of fortified water extracts 1 ppb, 2ppb, 5ppb, 1 ppb, 2 ppb, 5 ppb LOD = *RSD*[Conc.] RSD = STDEV(Peak Area) / Average(Peak Area)
60 USEPA Method LODs in Water Extract ppb Compounds <.5 ppb 2 ppb 21 5 ppb ppb 25 ppb Atrazine On Column [Conc.] in Water Extract XIC (2.15±.12) Da
61 USEPA Method LODs in Water Extract ppb Compounds <.5 ppb 2 ppb 21 5 ppb ppb 25 ppb Benzo[ghi]perylene On Column [Conc.] in Water Extract XIC (276.9±.16) Da
62 USEPA Method On Column L Sample <.5 ng/ml 2 ng/ml 21 5 ng/ml ng/ml 25 ng/ml <.5 ng/l 2 ng/l 21 5 ng/l ng/l 25 ng/l ng/ml = ppb ng/l = ppt
63 USEPA Method Concentration of organic compounds in a 1L river water sample Name CAS Formula R.T. (min) Quant Masses Peak S/N Conc. (ng/l) 2,4-Dimethylphenol C 8 H 1 O 4.52 XIC(17.6±.6) Diethyl Phthalate C 12 H 14 O XIC(222.9±.13) Atrazine C 8 H 14 ClN XIC(2.8±.12) Anthraquinone C 14 H 8 O XIC(28.5±.12) Aldrin C 12 H 8 Cl XIC(262.86±.16) Metolachlor C 15 H 22 ClNO XIC(162.14±.1) Fluoranthene C 16 H XIC(2.7±.12) Pyrene C 16 H XIC(22.8±.12) Chrysene C 18 H XIC(228.9±.14) Benzo[b]fluoranthene C 2 H XIC(252.9±.15)
64 Non-target Compounds in River Water Name CAS R.T. (min) Formula Area Similarity Peak S/N 2-Naphthalenol, 5,6,7,8-tetrahydro C 1 H 12 O 3.91E Dihydroactinidiolide C 11 H 16 O Diethyltoluamide C 12 H 17 NO ,6-Dibromohydroquinone C 6 H 4 Br 2 O Tris(1,3-dichloroisopropyl)phosphate C 9 H 15 Cl 6 O 4 P Dehydroabietic acid C 2 H 28 O Naphthalenol, 5,6,7,8-tetrahydro- Diethyltoluamide Dehydroabietic acid Dihydroactinidiolide 2,6-Dibromohydroquinone Tris(1,3-dichloroisopropyl)phosphate
65 Non-target Compounds in River Water Name CAS R.T. (min) Formula Area Similarity Peak S/N 2-Naphthalenol, 5,6,7,8-tetrahydro C 1 H 12 O 3.91E Dihydroactinidiolide C 11 H 16 O Diethyltoluamide C 12 H 17 NO ,6-Dibromohydroquinone C 6 H 4 Br 2 O Tris(1,3-dichloroisopropyl)phosphate C 9 H 15 Cl 6 O 4 P Dehydroabietic acid C 2 H 28 O
66 Non-target Compounds in River Water Name CAS R.T. (min) Formula Area Similarity Peak S/N 2-Naphthalenol, 5,6,7,8-tetrahydro C 1 H 12 O 3.91E Dihydroactinidiolide C 11 H 16 O Diethyltoluamide C 12 H 17 NO ,6-Dibromohydroquinone C 6 H 4 Br 2 O Tris(1,3-dichloroisopropyl)phosphate C 9 H 15 Cl 6 O 4 P Dehydroabietic acid C 2 H 28 O DEET
67 Made for Modern Day Challenges A better way: using the Pegasus BT GC-TOFMS Full scan data (1-15 m/z) Non-Target Deconvolution Excellent dynamic range Retrospective Analysis StayClean Ion Source Eliminate false positives Easily quantitate volatile and semi-volatile organic compounds
68 Made for Modern Day Challenges A better way: using the Pegasus BT GC-TOFMS Full scan data (1-15 m/z) Non-Target Deconvolution Excellent dynamic range Retrospective Analysis StayClean Ion Source Eliminate false positives Easily quantitate volatile and semi-volatile organic compounds
69 Made for Modern Day Challenges A better way: using the Pegasus BT GC-TOFMS Full mass range data (1-15 m/z) Non-Target Deconvolution Excellent dynamic range Retrospective Analysis StayClean Ion Source Eliminate false positives Easily quantitate volatile and semi-volatile organic compounds
70 Acknowledgements Eric Reiner Jack Cochran Brock Chittim Todd Richards, Joe Binkley, and the rest of the apps group at LECO
71 Take Your Analysis on the Road! LECO Bus Tour: California, January-February 217 Next Stop Colorado State University, April 26-27, 217 Plan your stop today!
72 For More Information Contact LECO at: World Headquarters/United States In United States: or Outside U.S.A.:
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