10 Secrets of Multiresidual Analysis - Agilent · The last components\ഠof gasoline take about 70...
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10 Secrets of Multiresidual Analysis Marco Zanotti Food Market Manager Agilent EMEAI
Transcript of 10 Secrets of Multiresidual Analysis - Agilent · The last components\ഠof gasoline take about 70...
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10 Secrets of Multiresidual Analysis
Marco Zanotti Food Market Manager Agilent EMEAI
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Multiresidual analysis
• Pesticides and other contaminants; • ~1000000 extimated analysis/year in EU; • 80-90% by Contract Labs; • GC-MSMS, LC-MSMS;
10 tips to increase: • robustness; • throughput; • coverage;
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#1 – Where to set the border
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LC GC
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GC-MS or LC-MS(/MS) - Which Technique is More Essential? Scherbaum E., Anastassiades M., Schüle. E., Bauer, N., Ellendt, K. and M. Wieland CRL for Pesticide Residues using Single Residue Methods hosted at the Chemisches und Veterinäruntersuchungsamt Stuttgart, Schaflandstr. 3/2, 70736 Fellbach, Germany EPRW 2008
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Target compounds (2 transitions each)
Current Routine Methods: • GC-QQQ # 380 • LC-QQQ # 250 Agilent «Records»: • GC-QQQ # 447 • LC-QQQ # 313
# Things worth considering: • Instrument not limiting factor; • Methods need maintenance;
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#2 – Sample Matrix: Friend or Foe?
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Slope Variation (%)
Green teaRed TeaBlack TeaChamomile
Matrix effects*
* Data by kind permission of Prof. Amadeo R. Fernández-Alba, EURL-FV, Almeria, Spain
LC/MS Analysis Increasing SUPPRESSION GC/MS Analysis
Increasing ENHANCEMENT
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Sample Matrix: Protectant Effect
10 Solvent Std Injections
10 Lettuce Matrix Std Injections
10 Satsuma Matrix Std Injections
10 Onion Matrix Std Injections
Repeat Solvent Std Injection
Solvent Std
50 ppb Ethion Peak Area in Solvent, Spiked Lettuce, Onion and Satsuma Extracts
120 Consecutive Injections over 54 Hours
# Matrix matched calibration prevents over-estimation
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2012 1980
#3 – Ultimate carrier gas technology
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Retention Time Locking
Original Method
After column maintenance
After column replacement
Different instrument
RTL enables control of one key compound ID parameter
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Capillary Flow Technology
Multiple detectors
Heart Cutting (Deans Switch)
QuickSwap
GCXGC
Backflush
Presenter
Presentation Notes
This is a list of some of the capabilities now made available by Agilent’s capillary flow technology. We will now go over some examples of these.
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Backflush
S/S Inlet
0.1 psi
45 psi MS
Aux EPC Split Vent Trap
Run
Post run
S/S Inlet
25 psi
4 psi MS
Aux EPC Split Vent Trap
Presenter
Presentation Notes
The TCEP column required in this method is not a bonded phase and has an upper temperature limit of 135 C. The last components of gasoline take about 70 minutes to elute from column 1. The oxygenates and aromatics of interest are finished eluting by 22 minutes. This means that about 50 min are wasted waiting for unimportant compounds to clean out from column 1. Because this system has EPC, as soon as the last analyte has eluted from column 2, the PCM module can be pressure programmed to a higher pressure (60 psi in this example) at the same time that the split/splitless inlet is programmed to a lower pressure (0.5 psi in this example). Now the flow in column 1 is reversed at 3.8 mL/min, backflushing the heavy material out through the split vent of the inlet. This backflushing reduced the analysis time from 70 min to less than 25 min.
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10% Fish Oil In Acetone: RTs with Backflushing
min5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1
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10 Runs without Backflushing: Retention times shift ~4-5 sec during 10 runs
min5.3 5.4 5.5 5.6 5.7 5.8 5.9 6 6.1 6.2
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# Retention Time Locking & Backflush a must!
Presenter
Presentation Notes
Note that without backflushing, the retention times shift after EACH INJECTION. Backflushing cures the problem. This is probably the single most convincing example of the importance of backflushing.
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#4 – Sample dilution
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The need for sensitivity
Solvent --
Black tea 38%
Cucumber 99%
Lemon 83%
Pepper 86%
Rocket 2%
Tomato 99%
Wheat 103% Recoveries
1 ppb Methomyl spike, 1µl injection
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The dilution effect
1 ppb Methomyl spike, 1µl injection – recoveries for dilution in Rocket
-- 2%
1:2 79%
1:5 89%
1:10 97%
Dilution & Recovery
1:2 1:5 1:10
# Match: • Instrument sensitivity
• Matrix complexity • Sample preparation
• 6420
• 6430 • x2
• 6460 • X8
• 6490 • x40
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#5 – Triple vs. High Resolution
MS/MS
Accurate Mass
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QQQ vs. Hi Res.
Tetrachlorvinphos, 1ppb
6490, triple 6550, QTOF
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QQQ vs. Hi Res.
Deltamethrin, 1ppb
6490, triple 6550, QTOF
Metribuzin, 1ppb
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Triple vs. High Resolution
Cheaper Easier to operate
Easier method set up Broader scope
# Hi Res suitable for multiresidue analysis
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#6 – Software rules
Data Analysis 27%
Collection 6%
Sample Analysis 6%
Sample Prep 61%
Customer Survey: % Time Spent for Analysis
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#6 – Software Rules
GC-MS
MassHunter
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Seamless Integration
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MassHunter and Multiresidual Analysis
Batch Table
Compound Information
(Chroms + Specs)
Calibration Curve +
Curve Fit Assistant
Compounds Sa
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Compounds
Sam
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Compounds
Outlier Flagging + Filtering
View Results from Single or Multiple compounds
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Compound-at-a-glance
Cal sample
# Leverage software and optimize reporting speed
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#7- Broadening the Scope
• Target Screening, ~ 300 compounds
• Target Screening, ~ 300 compounds • Database Screening, ~ 1000 compounds • Untargeted Screening, ~ ∞ compounds
Triple quad
Hi-Res
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Three approaches
Target MS EIC Target MS/MS EIC
Database Find by Formula algorithm
Molecular Feature Extraction Database Search
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Cpd 73: Methidathion: +ESI Scan (8.607, 8.637, 8.667 min, 3 Scans) Frag=120.0V Rucola_0_01.d
(M+H)+ 302.96845
(M+H)+ 304.96516
(M+H)+ 303.97113
(M H) 305 9688
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Counts vs. Mass-to-Charge (m/z)302.5 303 303.5 304 304.5 305 305.5 306 306.5
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Cpd 73: Methidathion: + FBF Spectrum (8.607-8.667 min) Rucola_0_01.d Subtract
(M+H)+ 302.96845
(M+NH4)+ 319.99558
(M+Na)+ 32
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Counts vs. Mass-to-Charge (m/z)302 304 306 308 310 312 314 316 318 320 322 324 326
Time
# Hi Res & the right Software broaden the scope
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#8 –All Ions
Accurate MS and MSMS... ... of ALL compounds!
Collision Energy
Low Energy – Collect Parent Ions
High Energy – Collect Fragments
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#8 – All Ions Acquisition
Low Energy Full Scan
High Energy Full Scan
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Methidathion C6H11N2O4PS3 + Product Ion Frag=120.0V [email protected] Kempe_Pest - import test.cdb
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Counts vs. Mass-to-Charge (m/z)40 60 80 100 120 140 160 180 200 220 240 260 280 300 320 340
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All Ions in practice – 2 spiked matrices
• Reliable fragment detection
• No false positives
Avocado LemonCompound name Blank 5 ppb 10 ppb 50 ppb Blank 5 ppb 10 ppb 50 ppb
Acephate q q q q q qAldicarb nq q q q qAzamethiphos q q q q qAzinphos-ethyl q q q nq q qAzinphos-methyl q q q q q qAzoxystrobin q q q q q qBenalaxyl q q q q q qBifenazate q q q q q qBispyribac q q q q q qBupirimate q q q q qCarbaryl q q q nq nq qCarbofuran nq nq q q q qCarboxin q q q nq q q qChlorfenvinphos q q q q q qChloroxuron q q q q q qClomazone q q q q q qCoumaphos q q q q q qCymiazole q q q nq q qDiazinon nq nq q q q qDiethofencarb nq q q q nq q q qDifenconazole q q q q q qDimethachlor q q q q q qDimethoate q q q q q qDimethomorph(E) nq q q nq q qDimoxystrobin q q q q q qEthidimuron q q q q q qEthion q q q q q qFenamiphos q q q q q qFenuron q q q q q qFlufenacet q q q q q qFlumetsulam q q q q q qFluquinconazole nq q q nq q qFlusilazol q q q q q qFlutriafol q q q nq nq qForchlorfenuron q q q nq q qHalofenozide q q q nq nq qImazalil q q q q qImidacloprid q q q q q qIndoxacarb q q q nq q qIprovalicarb q q q nq q qIsofenphos methyl q q q q q qIsoprothiolane q q q q q qLenacil q q q nq nq qMalaoxon q q q q q qMecarbam q q q q q qMepanipyrim q q q q q qMetalaxyl q q q q nq qMetazachlor q q q q nq qMethidathion q q q q q qMethoxyfenozide q q q nq q qMetobromuron q q q q q qMetolachlor q q q q q qMevinphos q q q q q qNitenpyram q q q q q qOmethoate q q q q q qOxadiazon nq q q q q qOxadixyl q q q q q qPencycuron q q q q qPhenthoate q q q nq q qPhosalone q q q q q qPhosmet q q q q q qPhosphamidon q q q q q qPicoxystrobin q q q q q qPirimifos-methyl q q q q q qProchloraz q q q q q qProfenofos q q q q q qPropamocarb q q q q q qPropoxur q q q q q qPropyzamide q q q nq nq qPyraclostrobin q q q q q qPyriproxyfen q q q q q qRotenone q q q nq nq qSpinosyn A q q q q q qSpirodiclofen q q q q qSpiromesifen q q q q q qSpiroxamine q q q q q qTebufenoxide q nq q q q qTepraloxydim q q q nq q qTerbumeton q q q nq nq qTetraconazol nq q q nq q qThiacloprid q q q q q qThiamethoxam q q q q q qTolylfluanide q q q q q qTriazophos q q q q q qTricyclazole q q q q q qTrietazine q q q q q qVamidothion q q q q q q
Automatic ID
Manual inspection
# All Ions: the new tool for Hi Res screening
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#9 – Don’t re-invent the wheel
Databases & Kits
• MS, MSMS,Accurate Mass • Certified Standards
Pesticide Analyzers
• Pre-configured • Factory Tested
Solutions • Custom made • From Sample Prep to
Validation
Customer CD
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#10 – No data comes from a broken instrument
# Always factor in support when selecting an instrument
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Summary
GC/LC QQQ Multiresidual Analysis >200/300 compounds
Hi Throughput Software
Hi Res suitable for Multiresidue
Broadening the Scope
Databases, Application Kits, Analyzers, Solutions
Instrument Reliability
GC Matrix Matched Calibration Backflush LC Sample Dilution All Ions
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Thanks to...
• Thomas Glauner • Chris Sandy • Joerg Riener • Laszlo Toelgyesi • Christoph Mueller • Juan Aybar • Jan Lembcke • Gordon Ross
