US EPA, Inert (other) Pesticide Ingredients in Pesticide ...
Minimize Method Development Time for Pesticide Screening ... · Pesticide Application Kit with...
Transcript of Minimize Method Development Time for Pesticide Screening ... · Pesticide Application Kit with...
Dr. Jerry Zweigenbaum
Senior Applications Scientist
Food Safety
Minimize Method
Development Time for
Pesticide Screening
Using LC/MS
Application Kit
LC/MS/MS Pesticide Analysis
October 5,2010Page 1
LC/MS/MS TRIPLE QUADRUPOLE
Pesticide Application Kit with Dynamic MRM
Database for the
LC/MS/MS Pesticide Analysis
October 5,2010Page 2
SampliQ
Extraction and
Dispersive SPE Kits
Agilent 1260 LC
Agilent 6460 LC/QQQ
Methods for YOUR Pesticides!!!!
LC/MS/MS Pesticide Analysis
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Copy YOUR list to the Dynamic MRM Database and
Search
LC/MS/MS Pesticide Analysis
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Search on the METHOD and the Names to get RT
You will Use
LC/MS/MS Pesticide Analysis
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Database is Directly Linked to OPTIMIZER
LC/MS/MS Pesticide Analysis
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Many Columns and Info but may be Hidden
LC/MS/MS Pesticide Analysis
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Test Mix Positive Ion Mode: Quick Start
Verification
LC/MS/MS Pesticide Analysis
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Test Mix Negative Ion Mode
LC/MS/MS Pesticide Analysis
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Dynamic MRM Acquisition Parameters –
Customized Methods from Database
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LC/MS/MS Pesticide Analysis
Dynamic MRM at the 10 pg level for 100 pesticides
1 min ΔR.T.- based acquisition window for each MRM transition
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LC/MS/MS Pesticide Analysis
Dynamic MRM 11 Compounds (22 MRMs) eluting in about 1
min window
Compound Name Precursor Ion Product Ion Retention Time
Cinosulfuron 414.1 183 5.579
Cinosulfuron (Q) 414.1 157 5.579
Chlorotoluron 213.1 72 5.642
Chlorotoluron (Q) 213.1 140 5.642
Atrazine 216.1 174 5.682
Atrazine (Q) 216.1 132 5.682
Carbaryl 202.1 145 5.736
Carbaryl (Q) 202.1 117 5.734
Carboxin 236.1 143 5.836
Carboxin (Q) 236.1 87 5.836
Chlorsulfuron 358.0 167 5.896
Chlorsulfuron (Q) 358.0 141 5.896
Ethiofencarb 226.1 107 5.937
Ethiofencarb (Q) 226.1 164 5.936
Dodemorph 282.3 116 6.073
Dodemorph (Q) 282.3 98 6.074
Diuron 233.0 72 6.101
Diuron (Q) 233.0 160 6.101
Cyprodinil 226.1 108 6.245
Cyprodinil (Q) 226.1 93 6.246
Difenoxurone 287.1 123 6.509
Difenoxurone (Q) 287.1 72 6.509
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LC/MS/MS Pesticide Analysis
Typical results: 10 pg Atrazine on-column using Dynamic MRM
6460A QQQ with Jet Stream Technology
•Avg. Signal Height: 15,650
•Avg. Signal Area: 50,966
•RSD: 3.2 %
•Estimated LOQ: 100 fg or less
•6-7 data points above FWHM
•3 sec FWHM, 6 sec @10% valley
•20 data points baseline-to-
baseline
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3 sec FWHM
10% valley
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LC/MS/MS Pesticide Analysis
Pesticides LC/MS method
Jet stream conditions
Drying gas temperature 275 ºC
Drying gas flow (nitrogen) 6 L/min
Nebulizer gas pressure (nitrogen) 35 psig
Capillary voltage 4000 V
Sheath gas temperature 325 ºC
Sheath gas flow 12 L/min
Nozzle voltage Off
6460A QQQ settings
MS1 and MS2 resolution Unit
Time Filtering peak width = 0.03 min
Dynamic MRM transitions up to 4000
Constant cycle time 373 ms
Delta EMV 400 V
1200 LC parameters
Column Eclipse Plus-C18, 2.1 x 100mm, 1.8 µm
Column temperature 35 ºC
Injection volume 5.0 mL
Autosampler temp 6 ºC
Needle wash flushport (MeOH:H2O 75:25), 5 secs
Mobile phase A = 0.01% formic acid in water
B =5 mM ammonium formate 0.01% formic acid in 95:5 acetonitrile:water
Flow rate 0.3 mL/min
Gradient 0 min 6% B
15 min 95% B
Stop time 20 min 6% B
Post time 1 min
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LC/MS/MS Pesticide Analysis
Dynamic MRM of 300 Pesticides two Transitions
Each
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LC/MS/MS Pesticide Analysis
Run with 1200-SL LC 2.1 x 100 C18 Eclipse PLUS 1.8 u
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LC/MS/MS Pesticide Analysis
Pesticide Mix run with Agilent 1290 Infinity 1260
Bar LC: Benefit of HIGHER PEAK CAPACITY
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LC 2.1 x 150 C18 Eclipse PLUS HD 1.8 u, 0.6 mL/min, 800 bar
October 5,2010
LC/MS/MS Pesticide Analysis
3x10
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Counts vs. Acquisition Time (min)0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5
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IS
MRM chromatograms of 50 ng/g fortified sample processed by EN method. Peak identification: 1. Acephate, 2. Pymetrozine, 3. Carbendazim, 4. Thiabendazole, 5. Imidacloprid, 6. Imazalil, 7. Propoxur, 8. Carbaryl, 9. Cyprodinil, 10. Ethoprophos, 11. Penconazole, 12. Kresoxim-methyl, IS: TPP.
Column Agilent Poroshell 120 EC-C18, 2.1 × 100 mm, 2.7 μm
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LC/MS/MS Pesticide Analysis
LC/MS/MS Pesticide Analysis
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DYNAMIC MRM – CREATE MY
OWN
Using YOUR Chromatography
LC/MS/MS Pesticide Analysis
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Building a Dynamic MRM Acquisition Method
LC/MS/MS Pesticide Analysis
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Create a Dynamic MRM method
Import the results generated in the custom report into acquisition method
Specify delta retention time window
Analyze medium level standard mixtures
fixed dwell time for each MRM transition using one time segment
MHW Quantitative Data Analysis gets retention times
Load tabular MRM from Database
Compound names, ISTD (optional)MRM transitions, fragmentor voltages,
collision energies
Building a Dynamic MRM Method – 4.00
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LC/MS/MS Pesticide Analysis
Select Datafile and Parameters
LC/MS/MS Pesticide Analysis
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Acquisition is Automatically Populated with
Retention Times and RT Windows
LC/MS/MS Pesticide Analysis
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Next View the Method
LC/MS/MS Pesticide Analysis
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LC/MS/MS Pesticide Analysis
View Method Provides Important Information in
Cohesive Format
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LC/MS/MS Pesticide Analysis
Finally – Set the Cycle Time
LC/MS/MS Pesticide Analysis
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Typical results: 10 pg Atrazine on-column using Dynamic MRM
6460A QQQ with Jet Stream Technology
•Avg. Signal Height: 15,650
•Avg. Signal Area: 50,966
•RSD: 3.2 %
•Estimated LOQ: 100 fg or less
•6-7 data points above FWHM
•3 sec FWHM, 6 sec @10% valley
•20 data points baseline-to-
baseline
•
•
•
•
•
•
••
•••
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•
• • •
•
•• • •
3 sec FWHM
10% valley
Page 29 October 5,2010
LC/MS/MS Pesticide Analysis
Summary
• Dynamic MRM Database provides quick method setup with
compound specific parameters and retention times
• Compounds not in method can be easily added
• New compounds can easily be added via optimizer to method
and database
• Custom methods can be easily developed and Dynamic MRM
readily created with “Update Method”
LC/MS/MS Pesticide Analysis
October 5,2010Page 30