Agilent 6470 Triple Quadrupole LC/MS System and EMR-Lipid. The Combination for Confident Pesticide Quantitation. Christoph Mueller
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1 Agilent 670 Triple Quadrupole LC/MS System and EMR-Lipid The Combination for Confident Pesticide Quantitation Christoph Mueller Agilent Technologies LCMS Application Scientist EMEAI Food Market Program Manager
2 Agenda 670 QQQ innovations and quantitation performance Experimental Sample Preparation LC-QQQ method Evaluation of Matrix Removal Evaluation of Pesticide Recovery
3 670 Innovations Agilent 90 Infinity II Multicolumn Thermostat Multisampler High Speed Pump Upgradeable to 69 QQQ with ifunnel Technology
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8 Experimental Samples Avocado (high lipid content) Spinach (low lipid content) Pesticides Test mix from Agilent Pesticides tmrm Application Kit (G7BA) with 0 pesticides + some herbizides for negative ion performance evaluation Spiked to samples before extraction to asses recovery 0 and 00 ng/g Spiked to extracts after extraction 0 and 00 ng/g Extraction EN QuEChERS extraction salt + PSA/C8 dspe (for high lipid content) EN QuEChERS extraction salt + EMR lipid dspe + EMR lipid polish salts
9 EMR vs QuEChERS workflow PSA/C8 g Avocado (0 g spinach) + STD, ml water, 0 ml ACN (% acetic acid) in 0 ml centrifuge tube, shake min EMR 0 g Avocado (0 g spinach) + STD, 0 ml ACN (% acetic acid) in 0 ml centrifuge tube. Add Bond Elut EN extraction salts, shake min Add Bond Elut EN extraction salts, shake for min Centrifuge (000 rpm, min) Centrifuge (000 rpm, min) Add ml water, then ml of ACN extract to EMR-Lipid dspe ml tube Transfer 6 ml to Bond Elut EN dispersive SPE ml tube (PSA/C8) Vortex min, centrifuge at,000 rpm for min. Transfer ml of supernatant to a EMR Lipid polish tube. Vortex min, centrifuge at,000 rpm for min. Vortex min, centrifuge at,000 rpm for min. 00 µl extract µl water in autosampler vial. (Spinach: 00 µl extract µl water) 00 µl extract µl water in autosampler vial.
10 UHPLC and QQQ Parameters UHPLC column Agilent Zorbax Eclipse Plus C8 RRHD. x 0 mm,.8 µm Mobile Phase A: water + mm ammonium acetate + 0.% acetic acid B: methanol + mm ammonium acetate + 0.% acetic acid Gradient program Flow rate Min ml/min %B Dynamic MRM acquisition with fast polarity switching (for matrix evaluation: precursor ion scan and MS scan)
11 AVOCADO MATRIX Evaluation of Matrix Removal
12 Precursor Ion Scan for Product Ion m/z 8 Phosphatidylcholin - Phospholipids = Group of compounds known for causing ion supression - Q Scan Collision C. Q fixed at m/z 8 x0 6 +ESI TIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (** -> **) Avocado EN acid 0.d.9.8 x0 6 +ESI TIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (** -> **) Avocado EN acid 0.d Counts vs. Acquisition Time (min) Counts vs. Acquisition Time (min)
13 Comparison of PC-(8:n/8:n) between EMR-Lipid and PSA/C8 Increasing level of saturation of fatty acid Increasing level of removal x0 +ESI EIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 ( , , , > ) Avocado EN acid 0.d.8.6 x0 +ESI EIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 ( , , , > ) Avocado EN acid 0.d OLPC PC(8:/8:) OOPC PC(8:/8:) LLPC PC(8:/8:) Counts vs. Acquisition Time (min) Counts vs. Acquisition Time (min)
14 Lyso-PC-(C8:n/0:0) Significantly smaller than PC, but FA saturation has the same effect on level of removal x0 6 +ESI EIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (8.000, 000,.000 -> ) Avocado EMR 0-r00.d Avocado EMR 0 [Fenpyroximat] x0. -> 66. Area= >. Area= Avocado EN acid 0 [Fenpyroximat] x0. -> 66. Area=967. ->. Area= Lyso-PC-(C8:/0:0) Lyso-PC-(C8:/0:0) Lyso-PC-(C8:/0:0) Counts vs. Acquisition Time (min)
15 Assessment of Total Extracted Matrix MS Scan, High Temperature and Extended Chromatography x0 7 +ESI TIC Scan Frag=.0V CF=0.000 DF=0.000 Avocado EN acid 0.d Diacylglycerols (+ some MGDG) Triacylglycerols??? Counts vs. Acquisition Time (min)
16 SPINACH MATRIX Evaluation of Matrix Removal
17 Precursor Ion Scan for Product Ion m/z 8 Phosphatidylcholin - Group of compounds known for causing ion supression - Q Scan Collision C. Q fixed at m/z 8 x0 6. +ESI TIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (** -> **) Avocado EN acid 0.d Avocado PSA/C8. x ESI TIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (** -> **) Spinach EN 0.d. Spinach PSA/C8. x ESI TIC Precursor Ion Frag=0.0V CF=0.000 DF=0.000 CID@0.0 (** -> **) Spinach EMR 0.d. Spinach EMR-Lipid Counts vs. Acquisition Time (min)
18 Assessment of Total Extracted Matrix MS Scan, High Temperature and Extended Chromatography x ESI TIC Scan Frag=.0V CF=0.000 DF=0.000 Avocado EN acid 0.d Avocado PSA/C8 x ESI TIC Scan Frag=.0V CF=0.000 DF=0.000 Spinach EN 0.d 6 Spinach PSA/C8 x ESI TIC Scan Frag=.0V CF=0.000 DF=0.000 Spinach EMR 0.d 6 Spinach EMR-Lipid Counts vs. Acquisition Time (min)
19 PESTICIDE RESULTS
20 Excellent performance at default MRL (0µg/kg) Pirimicarb
21 Signal Quality in region with large max. number of concurrent MRMs Uniconazole-P Fipronil Diflubenzuron Flusilazole Fenamiphos Picoxystrobin Bupirimate Fenoxycarb
22 Recovery of Pesticides from Avocado Matrix EMR Lipid vs QuEChERS 6 pesticides spiked EMR-Lipid PSA/C8 Reference for recovery is solvent calibration, which results in greater deviation from 70-0% range.
23 Lower Recovery with EMR-Lipid vs. PSA/C8
24 Lower Recovery in PSA/C8 vs. EMR-Lipid
25 Better Recovery with PSA/C8 Propamocarb Oxamyl Bifenazate Avocado EMR 0 [Propamocarb] Avocado EMR 0 [Oxamyl] Avocado EMR 0 [Bifenazate] x > 0. Area=7 x > 7. Area=66 x0 0. -> 98. Area= >.0 Area= > 90. Area=0 0. -> 70. Area= EMR Avocado EN acid 0 [Propamocarb] Avocado EN acid 0 [Oxamyl] Avocado EN acid 0 [Bifenazate] x > 0. Area=090 x > 7. Area=70 x0 0. -> 98. Area= >.0 Area= > 90. Area= > 70. Area= PSA/C
26 Better Recovery with PSA/C8 EMR Tribenuron-methyl Avocado EMR 0 [Tribenuron-methyl] x >. Area= > 8. Area= Avocado EMR 0 [Spirotetramat] x0 7. -> 0. Area= > 0. Area= Spirotetramat PSA/C8 0. Avocado EN acid 0 [Tribenuron-methyl] x >. Area= > 8. Area= Avocado EN acid 0 [Spirotetramat] x0 7. -> 0. Area= > 0. Area=
27 Better Recovery with EMR-Lipid Quinmerac Triclopyr Dichlorprop Avocado EMR 0 [Quinmerac] Avocado EMR 0 [Triclopyr] Avocado EMR 0 [Dichlorprop] x0.0 -> 0.0 Area=6098 x0.9 -> 9.9 Area=98 x0.0 -> 6.0 Area=.0 ->. Area= > 97.9 Area=97.0 -> 6.0 Area= EMR Avocado EN acid 0 [Quinmerac] Avocado EN acid 0 [Triclopyr] Avocado EN acid 0 [Dichlorprop] x0.0 -> 0.0 Area=60 x0.9 -> 9.9 Area=67 x0.0 -> 6.0 Area= >. Area=79.9 -> 97.9 Area=8.0 -> 6.0 Area=6.. 7 PSA/C
28 Better Recovery with EMR-Lipid,,-T Bispyribac Moxidectin Avocado EMR 0 [,,-T] Avocado EMR 0 [Bispyribac] Avocado EMR 0 [Moxidectin] x0.9 -> 9.9 Area=6 x0.0 -> 7. Area=7806 x0 *60. -> 6. Area=0.9 -> 8.9 Area=98.9 -> 96.9 Area=..0 ->. Area= > 8. Area= EMR PSA/C8 Avocado EN acid 0 [,,-T] x0.9 -> 9.9 Area=8.9 -> 8.9 Area=0.9 -> 96.9 Area= Avocado EN acid 0 [Bispyribac] x0.0 -> 7. Area=8.0 ->. Area= Avocado EN acid 0 [Moxidectin] x0 *60. -> 6. Area= > 8. Area=
29 Summary EMR-lipid clean-up dramatically reduces lipid content injected onto colum, while maintaining analyte recovery as with other dspe sorbents. EMR-lipid clean-up improves robustness of the method and longevity of column and mass spectrometer. Productivity gains through less maintenance downtime. Sensitive and robust 670 QQQ is perfect match for the analysis of diluted extracts after EMR-lipid clean-up. Thank You!
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