Application and evaluation of a high-resolution mass spectrometry screening method for veterinary drug residues in incurred fish and imported aquaculture samples
The ability to detect chemical contaminants, including veterinary drug residues in animal products such as fish, is an important example of food safety analysis. In this paper, a liquid chromatography high-resolution mass spectrometry (LC-HRMS) screening method using a quadrupole-Orbitrap instrument was applied to the analysis of veterinary drug residues in incurred tissues from aquacultured channel catfish, rainbow trout, and Atlantic salmon and imported aquacultured products including European eel, yellow croaker, and tilapia. Compared to traditional MS methods, the use of HRMS with nontargeted data acquisition and exact mass measurement capability greatly increased the scope of compounds that could be monitored simultaneously. The fish samples were prepared for analysis using a simple efficient procedure that consisted of an acidic acetonitrile extraction followed by solid phase extraction cleanup. Two different HRMS acquisition programs were used to analyze the fish extracts. This method detected and identified veterinary drugs including quinolones, fluoroquinolones, avermectins, dyes, and aminopenicillins at residue levels in fish that had been dosed with those compounds. A metabolite of amoxicillin, amoxicillin diketone, was also found at high levels in catfish, trout, and salmon. The method was also used to characterize drug residues in imported fish. In addition to confirming findings of fluoroquinolone and sulfonamide residues that were found by traditional targeted MS methods, several new compounds including 2-amino mebendazole in eel and ofloxacin in croaker were detected and identified.
KeywordsHigh-resolution mass spectrometry Screening method Veterinary drug residues
The authors would like to acknowledge chemists in the Denver Regulatory Laboratory who assisted in sample preparation and analysis, Dr. Cynthia Stine for assisting with generation of the incurred samples, as well as helpful discussions with Thermo Fisher scientists.
Compliance with ethical standards
Reference to any commercial materials, equipment, or process does not, in any way, constitute approval, endorsement, or recommendation by the U.S. Food and Drug Administration. In addition, the views expressed in this article are those of the author(s) and may not reflect the official policy of the Department of Health and Human Services, the U.S. Food and Drug Administration, or the U.S. Government.
The experimental protocol to generate the incurred samples was approved by the Animal Care and Use Committee at the FDA/CVM/OR, and all procedures were conducted in accordance with the principles stated in the Guide for the Care and Use of Laboratory Animals (2011) and the Animal Welfare Act of 1966 (P.L. 89-544), as amended.
Conflict of interest
The authors declare they have no conflict of interest.
- 1.World Bank Report. Fish to 2030: Prospects for Fisheries and Aquaculture. 2013.Google Scholar
- 5.US Government Accounting Office (GAO). Imported Seafood Safety: FDA and USDA could strengthen efforts to prevent unsafe drug residues. Report to the Chairman, Committee on Appropriations, US Senate. 2017;GAO-17-443.Google Scholar
- 6.Storey JM, Clark SB, Johnson AS, Andersen WC, Turnipseed SB, Lohne JJ, et al. Analysis of sulfonamides, trimethoprim, fluoroquinolones, quinolones, triphenylmethane dyes and methyltestosterone in fish and shrimp using liquid chromatography mass spectrometry. J Chromatogr B. 2014;972:38–47.CrossRefGoogle Scholar
- 11.Justino CIL, Duarte KR, Freitas A, C., Panteleitchouk TSL, Duarte AC, Rocha-Santols TAP. Contaminants in aquaculture: overview of analytical techniques for their determination. Trends Anal Chem 2016;80:293–310.Google Scholar
- 13.Turnipseed SB, Storey JM, Lohne JJ, Andersen WC, Burger RJ, Johnson AS, et al. Wide-scope screening method for multi-class veterinary drug residues in fish, shrimp, and eel using liquid chromatography-quadrupole high-resolution mass spectrometry. J Agric Food Chem. 2017;65:7252–67.CrossRefGoogle Scholar
- 16.FDA. Chemotherapeutics in Seafood Compliance Program 7304.018 2017.Google Scholar
- 17.FDA. Acceptance criteria for confirmation of identity of chemical residues using exact mass data for the FDA Foods and Veterinary Medicine Program. 2015.Google Scholar
- 18.Casey CR, Nickel T, Bradley JD, Ayres P. A rapid liquid chromatography-fluorescence detection for the quantitative analysis of avermectin residues in salmon and trout. FDA Laboratory Information Bulletin. 2018; in progress.Google Scholar
- 19.Casey CR, Karbiwnyk CM, Andersen WC, Ayres P. Liquid chromatography-tandem mass spectrometry method for the confirmation and quantitative analysis of avermectin residues in salmon. FDA Laboratory Information Bulletin. 2011;4496.Google Scholar
- 22.Roybal JE, Walker CC, Pfenning AP, Turnipseed SB, Storey JM, Gonzales SA, et al. Concurrent determination of four flouroquinolones in catfish, shrimp, and salmon by liquid chromatography with fluorescence detection. J AOAC Int. 2002;85:1293–301.Google Scholar
- 28.Verdon E, Andersen WC. Certain dyes as pharmacologically active substances in fish farming and other aquaculture products. In: Kay JF, MacNeil J, Wang J, editors. Chemical analysis of non-antimicrobial veterinary drug residues in food Hoboken. Avenel: Wiley; 2017. p. 497–531.Google Scholar
- 32.Wang J-H, Chao M-R, Chang M-H, Kuo T-F. Liquid chromatograph determination of amoxicillin residues in grouper muscle following oral administration of the veterinary drug. Taiwan Vet J. 2009;35:21–8.Google Scholar
- 33.Plakas SM, DePaola A, Moxey MB. Bacillus stearothermophilus disk assay for determining ampicillin residues in fish. J AOAC Int. 1991;74:910–12.Google Scholar
- 34.Wang B, Pang M, Xie X, Xie K, Zhang Y, Zhao X, et al. Quantitative analysis of amoxicillin, amoxicillin major metabolites, and ampicillin in chicken tissues via ultra-performance liquid chromatography-electrospray ionization tandem mass spectrometry. Food Anal Methods. 2017;10:3292–305.CrossRefGoogle Scholar
- 38.Ang CY, Luo W, Hansen EB Jr, Freeman JP, Thompson HC Jr. Determination of amoxicillin in catfish and salmon tissue by liquid chromatography with precolumn formaldehyde derivatization. J AOAC Int. 1996;79:389–96.Google Scholar
- 39.Andersen WC, Storey JM, Turnipseed SB, Wu I-L. unpublished work. 2018.Google Scholar
- 41.Iosifidou EG, Haagsma N, Olling M, Boon JH, Tanck MWT. Residue study of mebendazole and its metabolites hydroxy-mebendazole and amino-mebendazole in eel (Anguilla anguilla) after bath treatment. Drug Metab Dispos. 1997;25:317–20.Google Scholar
- 42.European Agency for the Evaluation of Medicinal Products (EMEA). Mebendazole Summary Report. EMEA/MRL/625/99-FINAL. 1999.Google Scholar
- 45.Wetzstein H-G, Schneider J, Karl W. Patterns of metabolites produced from the fluoroquinolone enrofloxacin by basidomycetes indigenous to agricultural sites. Applied Microbial and Cell PhysioloResearch in Veterinary Science. 2006;71:90–100.Google Scholar