Quantitative and confirmatory analysis of veterinary drug residues in food of animal origin by UPLC- MS/MS after QuEChERS clean-up
Michelle Whelan 1,2 Martin Danaher 1, Ambrose Furey 2. Ashtown Food Research Centre 1, C ork Institute of Technology 2. Topics Covered
• ProSafeBeef • Food Safety – Background – Anti-parasitic • Method Development – Existing method – New method – Sample preparation • Method Validation – Milk, Liver and Muscle • Method Performance • Future Work ProSafeBeef ProSafeBeef
• Advancing Beef Safety and Quality through Research and Innovation: European Framework Programme 6 • The ProSafeBeef consortium is multidisciplinary and comprised of 41 participants from research institutes, universities, private companies and industry organizations from 13 European countries, as well as Brazil, United States, Canada, Australia and New Zealand. • ProSafeBeef is divided into 7 separate management/research areas • Piller 1 - Quantitative Risk assessment for microbial and chemical contaminants Aim of Chemical Contaminants section 1.4 of Pillar 1
• Develop assays for anthelmintic residues • Transfer method to project partners, QUB, AFBI, IAEA, and Microbioticos • Inter-laboratory studies • Survey of EU beef (n = 1000 samples) • Exposure assessment • Quantitative risk assessment of chemical residues in beef • Dissemination Europe Climates Sampling Plan ProSafeBeef
• Important to identify the key anthelmintic drugs and their usage in different animal species • Also need knowledge of the time of application • Risk based approach is more likely to identify non-compliant results and will give greater consumer protection • Sampling is biased and may portray a negative image of food Food Safety Food Safety
• Main concern over benzimidazoles is due to their teratogenic properties (males and females) – Research has shown that OFZ causes birth defects if used in pregnant ewes • Secondary metabolite is generally the most toxic – Oxfendazole more toxic than fenbendazole – Hydroxy-mebendazole more toxic than mebendazole Background
• National Reference Laboratory for anthelmintic agents. • Veterinary drug residues are tested for at AFRC as part of the National Monitoring Plan. • Anthelmintics important in Ireland due to the extensive nature of farming • In early 2008: Four separate LC methods for anthelmintics in milk and liver (n=19 residues). • No method for flukicide and levamisole • Desired multi method for anthelmintics in milk, liver and muscle • Developed a multi method mid 2008 and demonstrated it at CRL in May 2009 Anthelmintics Anthelmintics drugs are used for controlling the following worms: Nematodes (roundworms) Cestodes (tapeworms) Trematodes (Flukes) Three main classes of drugs used for the treatment are: Benzimidazoles Avermectins Flukicides Economic losses Weight loss Poor wool growth and quality Reduced milk production Method development List of Analytes Abamectin Mebendazole Levamisole Doramectin Hydroxy Mebendazole Coumaphos Emamectin Amino Mebendazole Coumaphos-Oxon Eprinomectin Flubendazole Rafoxanide Ivermectin Hydroxy Flubendazole Oxyclozanide Moxidectin Amino Flubendazole Niclozamide Cambendazole Fenbendazole Nitroxinil Albendazole Fenbendazole Sulphone Bithionol Albendazole Sulphone Oxfendazole Clorsulon Albendazole Sulphoxide Triclabendazole Closantel Albendazole amino Oxibendazole Triclabendazole sulphone Sulphone 5 Hydroxy- Morantel Triclabendazole Thiabendazole Sulphoxide Thiabendazole Haloxon Existing LC-MS/MS method
• Quechers sample preparation was developed in USDA by Brian Kinsella • LC-MS/MS method was developed using Agilent 1100 and an Applied Biosystems API 3000 using a Prodigy C18 column, (150 x 3, 5 µm) • Two injections (pos and neg modes) 24 mins each. • Two internal standards • Only detects TCB and TCB-SO • LOQ 5ppb • Kinsella et al. Current UPLC-MS/MS method
• Modified sample preparation developed in USDA to detect lower levels – Scaled up Clean up step – Concentration step using DMSO • LC-MS/MS method was developed using Acquity UPLC and Waters MicroMass Quattro Premier XE using a HSS T3 column, (100 x 2.1 mm; 1.8 µm) • ONE injections (covers both pos and neg modes) 12.5 mins • 21 internal standards, 15 Deuterated • Detects all the TCB metabolites • LOQ < 2ppb UPLC Conditions
• Column: HSS T3 (100 x 2.1 Time A% B% Curve mm; 1.8 µm) 0.00 100 0 1 • Column Temp. : 60 °C 0.50 100 0 6 • MPA : 0.01% CH 3COOH in 5.00 50 50 6 H 0 + ACN (90 + 10, v/v) 2 7.00 10 90 6 • MPB : 5 mM Ammon. Form. in MeOH + ACN (75 + 25, v/v) 8.50 10 90 6 • Flow rate: 0.6 ml min -1 8.51 0 100 6 • Injection: 5 µl 9.50 0 100 6 • Run time: 12.5 min 9.51 100 0 6 12.50 100 0 6 Sample Preparation for non-MRL Substances
••ExtractionExtraction 10 g sample + 12 ml MeCN Homogenise (only for liver and muscle)
Add 4g MgSO 4 and 1g NaCl Shake for 1 minute and Centrifuge for 12 minutes ••CleanClean upup andand ConcentrationConcentration
Pour supernatant into centrifuge tube containing 1.5g MgSO 4 and 0.5g C 18 Mix for 30 seconds and Centrifuge for 10 minutes Transfer 6 mls to a test tube containing 250µl of DMSO Evaporate MeCN under nitrogen (60 °C) ••AnalysisAnalysis Filter the extracts and add to auto sampler vial. Inject Sample Preparation for MRL Substances
••ExtractionExtraction 10 g sample + 12 ml MeCN Homogenise (only for liver and muscle)
Add 4g MgSO 4 and 1g NaCl Shake for 1 minute and Centrifuge for 10 minutes ••CleanClean upup andand ConcentrationConcentration Add 1ml of the supernatant into micro-centrifuge tube containing 150mg MgSO 4 and 50mg C 18 Mix for 30 seconds and Centrifuge for 2 minutes ••AnalysisAnalysis Filter the extracts and add to auto sampler vial. Inject Method validation Method Validation
According to 2002/657/EC guidelines. Specificity was carried out using 20 blank samples. Linearity: > 0.98 Recovery Experiments: - 1, 1.5, 2 times the chosen level, (or) 0.5, 1, 1.5 times MRL Within laboratory repeatability with single analyst - 1, 1.5, 2 times the chosen level, (or) 0.5, 1, 1.5 times MRL Within laboratory reproducibility with three analysts - 1, 1.5, 2 times the chosen level, (or) 0.5, 1, 1.5 times MRL CC α and CC β values were calculated at 1, 1.5, 2 times the chosen level, or 0.5, 1, 1.5 times MRL. S/N ratio is typically >100 Ion Ratios very consistent and reproducible through-out runs. Recovery at validated levels for Muscle
160.0 1 1.5 2
140.0
120.0
100.0
80.0
60.0
40.0
20.0
0.0 34.0 32.0 Within Laboratory Reproducibility - Muscle - Benz 30.0 0.5 1 1.5 28.0
26.0
24.0
22.0
20.0
18.0
% CV % 16.0
14.0
12.0
10.0
8.0
6.0 4.0
2.0
0.0
e e e e e e le e e e e le n le le l le l le le l le le n le l id n o n o o o o o id o o o x o o o o o z z o o z o x o z z h h z z z h z a z z a z z h z a a o p p a a a a a a a a a o p a d h l l d d d lp d d d d d d d d h l d d p u u n u n n n n p u n n n l s s n n n e n n e n l s e e u - - e e fe -s e e b e e e e u - e b s e o b b b b b b b b b s e b b l - l n le u u lu e e i a - l a ia A e o in m Ox o l l -f e x l e o i h l z e z F -f M -m O c l z h t o m F o y -m ri o a T - z a a Ca a n x o y z y a d - d i n x T a d x n e n ro i d n o d e l e m d ro e r n b o b A m d n b d e l z n Hy A e a y lb A a Hy b l h d e la ic - A n F c r 5 e ri T lb T A 34.0 Within Laboratory Reproducibility - Muscle- Aver - Fluke 32.0 0.5 1 1.5 30.0
28.0
26.0
24.0
22.0
20.0
18.0
% CV % 16.0
14.0
12.0
10.0
8.0
6.0
4.0
2.0
0.0
l n l l il e n n le o e e e n e s n in in in in i in lo t t d id id o o o t t t t t t o n n n i y n h x x c c c c c c is io u a a x n p o o e e e e e e rs s r m o a a - l d m th o a tr z x a s m m m m i a i o o s o o o a a o rm x B M o l f m Ha ra e o v Cl Cl l Ni c u h b o m in v e ic y p A r I M L x Ra Co a D E p N O m E u Co Recovery at validated levels for Milk 180.0
160.0
140.0 1 1.5 2
120.0
100.0
80.0
60.0
40.0
20.0
0.0 32.0 Within Laboratory Reproducibility - Milk - Benz 30.0 28.0 0.5 1 1.5 26.0 24.0 22.0 20.0 18.0 16.0 14.0 % CV % 12.0 10.0 8.0 6.0 4.0 2.0 0.0 Within Laboratory Reproducibility - Milk - Fluke and 30.0
28.0 Aver
26.0 1 1.5 2
24.0
22.0
20.0
18.0
% CV % 16.0
14.0
12.0
10.0
8.0
6.0
4.0
2.0
0.0 Recovery at validated levels for Liver 160.0
1 1.5 2 140.0
120.0
100.0
80.0
60.0
40.0
20.0
0.0 30.0 Within Laboratory Reproducibility - Liver - Benz 28.0
26.0 1 1.5 2 24.0
22.0
20.0
18.0
16.0
14.0 % CV %
12.0
10.0
8.0
6.0
4.0
2.0
0.0 42.0 Within Laboratory Reproducibility - Liver - Flukever - A 40.0
38.0 1 1.5 2 36.0 34.0 32.0 30.0 28.0 26.0 24.0 22.0 20.0 % CV % 18.0 16.0 14.0 12.0 10.0 8.0 6.0 4.0 2.0 0.0 CC α and CC β for Milk milk LOR/MRL CC CC β µg/kg µg/kg Albendazole 100 107.40 116.61 Albendazole-sulphoxide 100 122.75 145.50 Albendazole-sulphone 100 104.77 115.75 Albendazole-amino-sulphone 100 104.52 119.96 Cambendazole 2 0.70 1.20 Fenbendazole 10 10.79 12.16 Oxfendazole 10 12.05 13.78 Fenbendazole-sulphone 10 10.84 11.81 Flubendazole 2 1.53 2.61 Amino-flubendazole 2 0.82 1.41 Hydroxy-flubendazole 2 0.80 1.36 Mebendazole 2 0.70 1.19 Amino-mebendazole 2 0.92 1.56 Hydroxy-mebendazole 2 0.69 1.17 Oxibendazole 2 1.95 3.32 Triclabendazole 2 0.71 1.20 Triclabendazole-sulphoxide 2 0.90 1.53 Triclabendazole-sulphone 2 1.33 2.26 Thiabendazole 2 1.65 2.81 5-hydroxy-thiabendazole 2 1.49 2.54 CC α and CC β for Milk Milk LOR/MRL CC CC β µg/kg Levamisole 2 1.26 2.14 Bithionol 4 1.59 2.71 Clorsulon 4 0.87 1.48 Closantel 2 1.07 1.82 Morantel 50 57.89 71.15 Niclosamide 2 0.74 1.27 Nitroxynil 2 0.66 1.12 Oxyclozanide 10 14.36 20.1 Rafoxanide 2 0.80 1.36 Coumaphos 2 0.82 1.40 Coumaphos-oxon 2 0.87 1.48 Haloxon 2 0.89 1.52 Abamectin 2 0.77 1.31 Doramectin 2 1.13 1.92 Emamectin 2 0.74 1.26 Eprinomectin 20 22.03 24.61 Ivermectin 2 0.74 1.25 Moxidectin 40 45.04 51.88 CC α and CC β for Bovine Liver Liver LOR/MRL CC CC β µg/kg µg/kg Albendazole 1000 1092.30 1173.85 Albendazole-sulphoxide 1000 1385.08 3844.29 Albendazole-sulphone 1000 1396.67 2403.42 Albendazole-amino-sulphone 1000 1051.59 1169.45 Cambendazole 2 0.32 0.55 Fenbendazole 500 552.81 620.57 Oxfendazole 500 537.20 589.62 Fenbendazole-sulphone 500 588.83 674.82 Flubendazole 2 0.87 1.49 Amino-flubendazole 2 0.87 1.49 Hydroxy-flubendazole 2 0.25 0.43 Mebendazole 2 0.21 0.36 Amino-mebendazole 2 0.87 1.49 Hydroxy-mebendazole 2 0.26 0.44 Oxibendazole 2 0.85 1.45 Triclabendazole 250 287.99 335.75 Triclabendazole-sulphoxide 250 438.55 1116.29 Triclabendazole-sulphone 250 1225.10 1825.22 Thiabendazole 2 0.39 0.66 5-hydroxy-thiabendazole 2 0.67 1.13 CC α and CC β for Bovine Liver Liver LOR/MRL CC CC β µg/kg Levamisole 2 0.30 0.52 Bithionol 4 0.43 0.74 Clorsulon 4 0.51 0.87 Closantel 2 0.45 0.77 Morantel 50 57.89 71.15 Niclosamide 2 0.59 1.00 Nitroxynil 2 0.38 0.65 Oxyclozanide 10 14.36 20.1 Rafoxanide 2 0.43 0.73 Coumaphos 2 0.63 1.07 Coumaphos-oxon 2 1.79 3.04 Haloxon 2 3.31 5.64 Abamectin 2 1.60 2.73 Doramectin 2 1.51 2.57 Emamectin 2 1.07 1.83 Eprinomectin 20 22.03 24.61 Ivermectin 2 1.09 1.86 Moxidectin 40 45.04 51.88 CC α and CC β for Muscle Muscle LOR/MRL CC CC β µg/kg µg/kg Albendazole 5 0.26 0.44 Albendazole-sulphoxide 5 0.54 0.91 Albendazole-sulphone 5 0.41 0.70 Albendazole-amino-sulphone 5 0.25 0.42 Cambendazole 5 0.24 0.40 Fenbendazole 5 0.26 0.44 Oxfendazole 5 0.34 0.58 Fenbendazole-sulphone 5 1.12 1.89 Flubendazole 5 0.51 0.86 Amino-flubendazole 5 0.40 0.67 Hydroxy-flubendazole 5 0.29 0.48 Mebendazole 5 0.26 0.44 Amino-mebendazole 5 0.58 0.98 Hydroxy-mebendazole 5 0.35 0.59 Oxibendazole 5 0.24 0.41 Triclabendazole 10 0.70 1.17 Triclabendazole-sulphoxide 10 2.38 4.02 Triclabendazole-sulphone 10 3.27 5.51 Thiabendazole 5 0.28 0.48 5-hydroxy-thiabendazole 5 0.50 0.85 CC α and CC β for Muscle Muscle LOR/MRL CC CC β µg/kg Levamisole 5 0.15 0.25 Bithionol 25 5.10 8.90 Clorsulon 25 10.21 17.23 Closantel 25 8.75 14.77 Morantel 25 4.66 7.87 Niclosamide 5 0.65 1.10 Nitroxynil 25 7.01 11.84 Oxyclozanide 10 1.99 3.37 Rafoxanide 5 1.72 2.90 Coumaphos 5 0.61 1.04 Coumaphos-oxon 5 0.33 0.56 Haloxon 5 0.84 1.41 Abamectin 5 1.09 1.85 Doramectin 5 1.05 1.77 Emamectin 5 0.92 1.55 Eprinomectin 5 0.69 1.17 Ivermectin 5 1.92 3.22 Moxidectin 5 2.57 4.31 Year 1 Survey Results Survey Results Year 1 - PSB EU Beef very safe 500 beef steaks 97.78% 97.78% of samples 2.22% residue free No risk to the consumer
1.22% -> Clos 0.20% -> Iver 0.20% -> Abz 0.20% -> Moxi 0.20% -> Eprino 0.20% -> Leva
Iver 1 sample non-compliant contained ivermectin (no-MRL) Method performance LC Versus UPLC-MS/MS for Milk HPLC Analysis UPLC-MS/MS Time (36 samples) (36 samples) No. analytes 19 38
No. runs 4 1
Sample Prep. (days) 6 1 Run time (min) 68.1 (33 + 35.1) 12.5
No. injections 144 64
Analysis time (h) 81.8 13.3
Processing time (days) 3 1
Totals (days) 12.5 2.5 Application to incurred milk
Sample No. Analyte Zu score
1 Blank -
2 Sum ABZ -0.698
3 Sum FBZ -0.927
4 Sum TCB +0.730
5 Levamisloe -0.019
6 Levamisle +0.570 Sum TCB +0.198 Application to incurred muscle Provisional Results
Sample No. Analyte Zu score
1 Blank -
2 Sum ABZ -1.001 Sum TCB +0.409 3 Sum ABZ -0.230 Sum MBZ -0.095 4 Levamisole -0.140 Sum FBZ -0.869 Sum TCB +0.129 5 Levamisole -0.010 Sum FBZ -0.346 6 Sum FBZ -0.040 Sum MBZ -0.254 Overlay of 38 Analytes 2ppb Overlay of Internal Standards ABZ and metabolites 2 ppb
Pos Ctl 2 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 6: MRM of 19 Channels ES+ 5.54 100 266.07 > 191.03 (ABZ) 44106 9.71e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 6: MRM of 19 Channels ES+ 5.54 100 266.07 > 234 (ABZ) 35989 8.09e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 5: MRM of 16 Channels ES+ 100 298.1 > 266.2 (ABZ-SO2) 1.33e6 3.61 Area % 23364 0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 5: MRM of 16 Channels ES+ 100 3.61 298.1 > 159.08 (ABZ-SO2) 29136 4.83e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 5: MRM of 16 Channels ES+ 3.31 100 282.24 > 240.1 (ABZ-SO) 19838 2.94e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 5: MRM of 16 Channels ES+ 100 3.31 282.24 > 159.06 (ABZ-SO) 8797 1.36e5 Area %
0 Time 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 Closantel & Rafoxanide 2ppb
Pos Ctl 2 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 9: MRM of 22 Channels ES- 100 6.83 660.85 > 315.1 (Closantel) 2415 7.24e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 9: MRM of 22 Channels ES- 100 6.83 660.84 > 126.9 (Closantel) 4438 1.38e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 9: MRM of 22 Channels ES- 100 7.09 623.79 > 344.83 (Rafox) 3050 8.40e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 9: MRM of 22 Channels ES- 100 7.09 623.79 > 126.87 (Rafox) 3673 9.52e4 Area %
0 Time 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 Clorsulon 4 ppb, Nitroxynil 2 ppb
Pos Ctl 2 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 4: MRM of 5 Channels ES- 3.33 100 379.8 > 343.95 (Clorsulon) 664 1.02e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 4: MRM of 5 Channels ES- 3.33 100 377.7 > 341.95 (Clorsulon) 1028 1.60e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 4: MRM of 5 Channels ES- 3.22 100 288.9 > 161.95 (Nitrox) 6684 9.86e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 4: MRM of 5 Channels ES- 3.22 100 288.9 > 126.86 (Nitrox) 6885 9.95e4 Area %
0 Time 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 Avermectins 2ppb
Pos Ctl 2 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.86 916.6 > 593.35 (Dora) 561 1.84e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.86 916.6 > 331.3 (Dora) 816 2.72e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.57 915.15 > 298.15 (Eprino) 387 2.02e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.57 915.15 > 144.06 (Eprino) 493 2.23e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.31 886.54 > 158.01 (Ema) 78444 3.26e6 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.31 886.54 > 126.05 (Ema) 4603 1.98e5 Area %
0 Time 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 Avermectins 2 ppb
Pos Ctl 2 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 7.83 100 640.25 > 528.4 (Moxi) 4970 1.77e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.83 640.25 > 498.3 (Moxi) 3129 1.15e5 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 8.16 100 892.25 > 569.45 (Iver) 1658 4.67e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 8.16 100 892.25 > 307.35 (Iver) 1560 4.14e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 100 7.67 890.5 > 567.1 (Aba) 1037 4.31e4 Area %
0 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 2009Apr21_milkval _CCa_HC_06 Sm (SG, 1x3) 10: MRM of 13 Channels ES+ 7.68 100 890.5 > 305.15 (Aba) 1406 5.47e4 Area %
0 Time 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00 10.00 Future Work
Year 2 samples ProSafeBeef Risk Assessment from analysis of beef samples (RIKILT) Improve precision for residues with CV >23% Compare QuEChERS against reference methods using incurred samples Stability study in solvent and Matrix Acknowledgements • Mr. Brian Kinsella (AFRC) • Dr. Helen Cantwell (AFRC) • Dr. Steve Lehotay (USDA, Wyndmoor) • This research was supported by EU Framework VI programme on Food Quality and Safety, ProSafeBeef project FOOD-CT-2006-36241