124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Veterinary Drug Residues

124th AOAC Annual Meeting & ExpositionSeptember 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



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.”



Call for seafood methods

Drug residues of interest:

Drug Residues Examples Analytical Range (µg/g or ppb)

Nitrofuran marker residues

SEM, AHD, AMOZ, AOZ 1.0-3.6

Chloramphenicol 0.3-1.2

Fluoroquinolones Ciprofloxacin, enrofloxacin, sarafloxacin

1.0-3.6

Quinolones Oxolinic acid, flumequine, naladixic acid

0.5-2.0

Dyes Malachite green, crystal violet, leuco forms

1.0-3.6

Methyltestosterone 0.8-3.0



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



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



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.

Title From Extraction/Clean-up Detection Detection LimitsRecoveries(RSDs)

Other Comments

Multiresidue method for the triphenylmethane dyes in fish: MG, CV, and BG(catfish, trout, basa, tilapia, salmon, shrimp)

FDADEN/ADRC

Extract w/ ammonium acetate buffer (pH 4.5) + ACN, (HAH, p-TSA) (2x) LLE w/ DCM and H2O

(2x) DDQ oxidation (leuco --> dye) SPE (Alumina N + PRS, stacked, 1 step)

LC & LC-MS/MSLC:C18 ; ACN/ AmmAcVIS:620 (MG); 588 (GV)LC-MS :LC: PH ; ACN/0.1%FMS : ND-APCI+, MS2

LODs: 0.1-0.2 ppb LOQs: 0.2-0.5 ppb > 86% (11%) MG > 91% (11%) CV 67% (15%) BGValidation: n

>30/cmpd+ incurred samples

Cmpds:LMG+MG, LCV+CV, BG Quant:-external stds (VIS)Confirm: MS2 spectra



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):



Recent trends in vet drug residue methods

Multiclass multiresidue methods

Streamlined sample preparation

Accurate mass LC-MS



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.



Streamlined sample preparation: QuEChERS


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.



Streamlined sample prep: On-line SPE


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.



Accurate mass LC-MS Targeted & non-targeted analytes TOF, Q-TOF Orbitrap

Several symposia at the meeting feature this topic



Accurate mass LC-MSExamples 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.



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

Documents

124th AOAC Annual Meeting & Exposition September 26-29, 2010 Loews Royal Pacific Resort at Universal Orlando Orlando, Florida Veterinary Drug Residues