1. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Veterinary Drug Residues Subgroup Report for 2010 AOAC Chemical Contaminants & Residues in Food Community

2. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Call for seafood methods Call for methods Drug Residues in Shrimp, Catfish, Tilapia, and Salmon “Validated multi-residue method(s) for select veterinary drug residues using high technology in shrimp, catfish, tilapia, and salmon has been identified as method(s) that are most urgently needed. AOAC INTERNATIONAL is inviting method authors to submit your method to AOAC for consideration and evaluation through the AOAC Official Methods program.”

3. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Call for seafood methods Drug residues of interest: Drug ResiduesExamplesAnalytical Range (µg/g or ppb) Nitrofuran marker residues SEM, AHD, AMOZ, AOZ1.0-3.6 Chloramphenicol0.3-1.2 FluoroquinolonesCiprofloxacin, enrofloxacin, sarafloxacin 1.0-3.6 QuinolonesOxolinic acid, flumequine, naladixic acid 0.5-2.0 DyesMalachite green, crystal violet, leuco forms 1.0-3.6 Methyltestosterone0.8-3.0

4. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Response to call for methods In response to this call, there were over 20 submissions: 5 for fluoroquinolones and quinolones 5 for phenicols (chloramphenicol) 6 for dyes 5 for nitrofurans 0 for methyltestosterone 4 multiclass multiresidue methods  Methods submitted from state, federal, private labs  Methods from 10 different countries to be evaluated  Several other applicable methods found in the literature

5. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Subgroup to evaluate methods Subgroup meeting tomorrow (Tuesday) from 1:15 – 3:45 PM Evaluate seafood methods Recommend 2-3 methods of each type (multi, NFs, etc.) Identify potential study directors and participating labs Discussion may be continued via WebEx

6. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Subgroup to evaluate methods Methods have been summarized: Summary tables will be available at the subgroup meeting. Copies of full submissions will also be available. Subgroup attendees will need to sign confidentiality agreement. TitleFromExtraction/Clean-upDetectionDetection Limits Recoveries(RSDs) Other Comments Multiresidue method for the triphenylmethane dyes in fish: MG, CV, and BG (catfish, trout, basa, tilapia, salmon, shrimp) FDA DEN/ ADRC Extract w/ ammonium acetate buffer (pH 4.5) + ACN, (HAH, p- TSA) (2x) LLE w/ DCM and H 2 O (2x) DDQ oxidation (leuco --> dye) SPE (Alumina N + PRS, stacked, 1 step) LC & LC-MS/MS LC:C18 ; ACN/ AmmAc VIS:620 (MG); 588 (GV) LC-MS : LC: PH ; ACN/0.1%F MS : ND-APCI +, MS 2 LODs: 0.1-0.2 ppb LOQs: 0.2-0.5 ppb > 86% (11%) MG > 91% (11%) CV 67% (15%) BG Validation: n >30/cmpd + incurred samples Cmpds:LMG+MG, LCV+CV, BG Quant:-external stds (VIS) Confirm: MS 2 spectra

7. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Other AOAC business relating to veterinary drugs  2009: Under contract with Elanco Animal Health, an AOAC ERP met to evaluate methods for tylosin, tilmicosin, narasin, monensin, and ractopamine in animal tissue.  2010: Contract was extended to nicarbazin, avilamycin, and apramycin. A call for methods has been issued, and another ERP will meet to evaluate procedures for these drug residues. Expert Review Panels (ERPs):

8. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Recent trends in vet drug residue methods  Multiclass multiresidue methods  Streamlined sample preparation  Accurate mass LC-MS

9. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Multiclass multiresidue methods Examples of recent publications: Multiclass, multiresidue drug analysis, including aminoglycosides, in animal tissue using LC coupled to tandem MS Martos et al., J. Agric. Food Chem. 2010, 58, 5932. Multiclass, multiresidue method for the detection of antibiotic residues in distillers grains by LC and ion trap tandem MS. De Alwis & Heller, J. Chromatogr. A 2010, 1217, 3076.

10. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Streamlined sample preparation: QuEChERS Examples of recent publications:  Development of fast screening methods for the analysis of veterinary drug residues in milk by LC-triple quadrupole MS. Vidal, et al., Anal. Bioanal. Chem. 2010, 397, 2777.  The development and validation of a multiclass liquid chromatography tandem LC-MS/MS procedure for the determination of veterinary drug residues in animal tissue using a QuEChERS approach. Stubbings & Bigwood, Anal. Chim. Acta 2009, 637, 68.

11. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Streamlined sample prep: On-line SPE Examples of recent publications:  Fully automated screening of veterinary drugs in milk by turbulent flow chromatography and tandem MS. Stolker, et al., Anal. Bioanal. Chem. 2010, 397, 2841.  High-throughput screening for multi-class veterinary drug residues in animal muscle using LC/tandem MS with on-line solid-phase extraction. Tang, et al., Rapid Commun. Mass Spectrom. 2006, 20, 2565.

12. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Accurate mass LC-MS  Targeted & non-targeted analytes  TOF, Q-TOF  Orbitrap Several symposia at the meeting feature this topic

13. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Accurate mass LC-MS Examples of recent publications:  Multi-residue screening of veterinary drugs in egg, fish and meat using high resolution LC accurate mass TOF MS. Peters, et al., J. Chromatogr. A 2009, 1216, 8206.  Comprehensive fast multiresidue screening of 150 veterinary drugs in milk by UHPLC coupled to TOF MS. Ortelli, et al., J. Chromatogr. B 2009, 877, 2363.  Full-Scan accurate Mass Selectivity of UHPLC Combined with TOF and Orbitrap MS in Hormone and Veterinary Drug Residue Analysis. van der Heeft, et al., J. Amer. Soc. Mass Spectrom. 2009, 20, 451.

14. 124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Veterinary Drug Residues Subgroup Reminder: Subgroup meeting tomorrow (Tuesday) from 1:15 – 3:45 PM Evaluate seafood methods Any other issues members want to discuss