Food Analytical Methods

, Volume 11, Issue 6, pp 1815–1831 | Cite as

Multi-residue Determination of Veterinary Drugs in Fishery Products Using Liquid Chromatography-Tandem Mass Spectrometry

  • Dasom Shin
  • Hui-Seung Kang
  • Jiyoon Jeong
  • Joohye Kim
  • Won Jo Choe
  • Kwang Soo Lee
  • Gyu-Seek Rhee


The aim of the present study was to develop a multi-residue confirmation method for the analysis of veterinary drugs in fishery products using liquid chromatography-tandem mass spectrometry. The chosen veterinary drugs, which are legally authorized compounds in Korea, were categorized into chemical classes such as amphenicols, cephalosporins, fluoroquinolones, macrolides, penicillins, pleuromutilins, sulfonamides, and tetracyclines. Sample extraction and cleanup were evaluated based on a modified quick, easy, cheap, effective, rugged, and safe procedure using octadecylsilane and primary secondary amine sorbents for purification. Extractions were carried out using an ammonium acetate buffer and ethylenediaminetetraacetic acid disodium salt (pH 4.0) with the addition of ammonium formate in water/acetonitrile. Instrumental analysis was performed using positive/negative ionization in the multiple reaction monitoring mode. The chromatographic conditions were optimized to give run time of < 20 min, and the developed method was validated according to the Codex Alimentarius Commission guidelines. Mean recoveries of the veterinary drugs ranged from 68.1 to 111%. At all target concentrations, the intraday precision was < 15%, and the lowest limits of detection and quantification were < 5 and < 10 μg kg−1. To evaluate the imprecision arising from residual matrix components, the linearity of our method was determined as > 0.98 using matrix-matched calibrations constructed at six different concentrations. The proposed method was applied to the analysis of 403 real samples obtained from domestic markets in Korea, and a number of samples tested positive (27.1%) for the targeted drugs. This analytical method is suitable for reliable detection of veterinary drug residues in fishery products.


Multi-residue determination Veterinary drug Maximum residue limit Fishery product Liquid chromatography-tandem mass spectrometry 



This work was supported by the Ministry of Food and Drug Safety of Korea.


This study was funded by the Ministry of Food and Drug Safety of Korea (Grant numbers 16161MFDS602 and 17161MFDS651).

Compliance with Ethical Standards

Not applicable

Conflict of Interest

Dasom Shin declares that she has no conflict of interest. Hui-Seung Kang declares that he has no conflict of interest. Jiyoon Jeong declares that she has no conflict of interest. Joohye Kim declares that she has no conflict of interest. Won Jo Choe declares that he has no conflict of interest. Kwang Soo Lee declares that he has no conflict of interest. Gyu-Seek Rhee declares that she has no conflict of interest.

Research Involving Human Participants and/or Animals

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed Consent

Not applicable

Supplementary material

12161_2018_1179_MOESM1_ESM.docx (233 kb)
ESM 1 (DOCX 232 kb)


  1. Aguilera-Luiz MM, Martinez Viedal JL, Romero-Gonzalez R, Garrido Frenich A (2012) Multiclass method for fast determination of veterinary drug residues in baby food by ultra-high-performance liquid chromatography-tandem mass spectrometry. Food Chem 132(4):2171–2180. CrossRefGoogle Scholar
  2. Anderson CR, Rupp HS, Wu W-H (2005) Complexities in tetracycline analysis—chemistry, matrix extraction, cleanup, and liquid chromatography. J Chromatogr A 1075(1-2):23–32. CrossRefGoogle Scholar
  3. Biselli S, Schwalb U, Meyer A, Hartig L (2013) A multi-class, multi-analyte method for routine analysis of 84 veterinary drugs in chicken muscle using simple extraction and LC-MS/MS. Food Addit Contam Part A 30(6):921–939. CrossRefGoogle Scholar
  4. Bousova K, Senyuva H, Mittendorf K (2013) Quantitative multi-residue method for determination antibiotics in chicken meat using turbulent flow chromatography coupled to liquid chromatography–tandem mass spectrometry. J Chromatogr A 1274:19–27. CrossRefGoogle Scholar
  5. CAC Codex Alimentarius Commission (2015) 38th Session of the Codex Veterinary Drug Residue in Food Online Database [Internet] Retrieved from :
  6. CCRVDF Codex Committee on Residues of Veterinary Drugs in Foods (2012) Guidelines for the design and implementation of national regulatory food safety assurance programme associated with the use of veterinary drugs in food producing animals (CAC/GL 71–2009) Rome: Codex Alimentarius Commission (CAC); p. 1–30Google Scholar
  7. Chambers E, Wagrowski-Diehl DM, Lu Z, Mazzeo JR (2007) Systematic and comprehensive strategy for reducing matrix effects in LC/MS/MS analyses. J Chromatogr B 852(1-2):22–34. CrossRefGoogle Scholar
  8. Chatterjee NS, Utture S, Banerjee K, Ahammed Shabeer TP, Kamble N, Mathew S, Ashok Kumar K (2016) Multiresidue analysis of multiclass pesticides and polyaromatic hydrocarbons in fatty fish by gas chromatography tandem mass spectrometry and evaluation of matrix effect. Food Chem 196:1–8. CrossRefGoogle Scholar
  9. Chen J, Korfmacher WA, Hsieh Y (2005) Chiral liquid chromatography-tandem mass spectrometric methods for stereoisomeric pharmaceutical determinations. J Chromatogr B 820(1):1–8. CrossRefGoogle Scholar
  10. Chen D, Yu J, Pan Y, Xie S, Huang L, Peng D, Wang X, Wang Y, Liu Z, Yuan Z (2016) Qualitative screening of veterinary anti-microbial agents in tissues, milk, and eggs of food-producing animals using liquid chromatography coupled with tandem mass spectrometry. J Chromatogra B 1017-1018:82–88. CrossRefGoogle Scholar
  11. Dasenaki ME, Thomaidis NS (2015a) Multi-residue determination of 115 veterinary drugs and pharmaceutical residues in milk powder, butter, fish tissue and eggs using liquid chromatography–tandem mass spectrometry. Anal Chim Acta 880:103–121. CrossRefGoogle Scholar
  12. Dasenaki ME, Thomaidis NS (2015b) Multianalyte method for the determination of pharmaceuticals in wastewater samples using solid-phase extraction and liquid chromatography–tandem mass spectrometry. Anal Bioanal Chem 407(15):4229–4245. CrossRefGoogle Scholar
  13. Díaz-Cruz MS, Barceló D (2006) Determination of antimicrobial residues and metabolites in the aquatic environment by liquid chromatography tandem mass spectrometry. Anal Bioanal Chem 386(4):973–985. CrossRefGoogle Scholar
  14. Dubreil E, Gautier S, Fourmond MP (2017) Validation approach for a fast and simple targeted screening method for 75 antibiotics in meat and aquaculture products using LC-MS/MS. Food Addit Contam Part A 34(4):453–468. CrossRefGoogle Scholar
  15. EC (2010) Council Regulation (EC) 37/2010/EC of 22 December 2009 on pharmacologically active substances and their classification regarding maximum residue limits in foodstuffs of animal origin (2010). Off J Eur Communities L15:1–72Google Scholar
  16. Fifield FW, Haines PJ (2000) Environmental analytical chemistry Blackwell Publishing pp. 4–5. ISBN 0-632-05383-6Google Scholar
  17. Geis-Asteggiante L, Lehotay SJ, Lightfied AR, Dutko T, Ng C, Bluhm L (2012) Ruggedness testing and validation of a practical analytical method for > 100 veterinary drug residues in bovine muscle by ultrahigh performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 1258:43–54. CrossRefGoogle Scholar
  18. Gomez-Perez ML, Plaza-Bolanos P, Romero-Gonzalez R, Martinez-Vidal JL, Garrido-Frenich (2012) Comprehensive qualitative and quantitative determination of pesticides and veterinary drugs in honey using liquid chromatography-Orbitrap high resolution mass spectrometry. J Chromatogr A 1248:130–138. CrossRefGoogle Scholar
  19. González-Curbelo MÁ, Lehotay SJ, Hernández-Borges J, Rodríguez-Delgado MÁ (2014) Use of ammonium formate in QuEChERS for high-throughput analysis of pesticides in food by fast, low-pressure gas chromatography and liquid chromatography tandem mass spectrometry. J Chromatogr A 1358:75–84. CrossRefGoogle Scholar
  20. Guan W, Li Z, Zhang H, Hong H, Rebeyev N, Ye Y, Ma Y (2013) Amine modified graphene as reversed-dispersive solid phase extraction materials combined with liquid chromatography–tandem mass spectrometry for pesticide multi-residue analysis in oil crops. J Chromatogr A 1286:1–8. CrossRefGoogle Scholar
  21. Johnson D, Boyes B, Orlando R (2013) The use of ammonium formate as a mobile-phase modifier for LC-MS/MS analysis of trypric digests. J Biomol Tech 24(4):187–197. CrossRefGoogle Scholar
  22. Kang HS, Lee SB, Shin D, Jeong J, Hong JH, Rhee GS (2018) Occurrence of veterinary drug residues in farmed fishery products in South Korea. Food Control 85:57–65. CrossRefGoogle Scholar
  23. Li C, Jin F, Yu Z, Qi Y, Shi X, Wang M, Shao H, Jin M, Wang J, Yang M (2012) Rapid determination of chlormequat in meat by dispersive solid-phase extraction and hydrophilic interaction liquid chromatography (HILIC)–electrospray tandem mass spectrometry. J Agric Food Chem 60(27):6816–6822. CrossRefGoogle Scholar
  24. Lopes RP, Reyes RC, Romero-González R, Frenich AG, Vidal JLM (2012a) Development and validation of a multiclass method for the determination of veterinary drug residues in chicken by ultra high performance liquid chromatography–tandem mass spectrometry. Talanta 89:201–208. CrossRefGoogle Scholar
  25. Lopes RP, Reyes RC, Romero-González R, Vidal JLM, Frenich AG (2012b) Multiresidue determination of veterinary drugs in aquaculture fish samples by ultra high performance liquid chromatography coupled to tandem mass spectrometry. J Chromatogr B 895–896:39–47. CrossRefGoogle Scholar
  26. Love DC, Rodman S, Neff RA, Nachman KE (2011) Veterinary drug residues in seafood inspected by the European Union, United States, Canada, and Japan from 2000 to 2009. Environ Sci Technol 45(17):7232–7240. CrossRefGoogle Scholar
  27. Mallet CR, Lu Z, Mazzeo JR (2004) A study of ion suppression effects in electrospray ionization from mobile phase additives and solid-phase extracts. Rapid Commun Mass Spectrom 18(1):49–58. CrossRefGoogle Scholar
  28. Masiá A, Suarez-Varela MM, Llopis-Gonzalez A, Picó Y (2016) Determination of pesticides and veterinary drug residues in food by liquid chromatography-mass spectrometry: a review. Anal Chim Acta 936:40–61. CrossRefGoogle Scholar
  29. Ministry of Food and Drug Safety (MFDS) (2015) Available from:
  30. Ministry of Food and Drug Safety (MFDS) (2017) Korean food standard code. Osong, KoreaGoogle Scholar
  31. Niessen WMA, Manini P, Andreoli R (2006) Matrix effects in quantitative pesticide analysis using liquid chromatography–mass spectrometry. Mass Spectrom Rev 25(6):881–899. CrossRefGoogle Scholar
  32. Picó Y, Font G, Ruiz MJ, Fernández M (2006) Control of pesticide residues by liquid chromatographymass spectrometry to ensure food safety. Mass Spectrometry Rev 25(6):17–960. Google Scholar
  33. Robert C, Gillard N, Brasseur PY, Pierret G, Ralet N, Dubois M, Delahaut P (2013) Rapid multi-residue and multi-class qualitative screening for veterinary drugs in foods of animal origin by UHPLC-MS/MS. Food Addit Contam Part A 30(3):443–457. CrossRefGoogle Scholar
  34. Romero-González R, López-Martínez JC, Gómez-Milán E, Garrido-Frenich A, Martínez-Vidal JL (2007) Simultaneous determination of selected veterinary antibiotics in gilthead seabream (Sparus Aurata) by liquid chromatography–mass spectrometry. J Chromatogr B 857(1):142–148. CrossRefGoogle Scholar
  35. Schmitz-Afonso I, Loyo-Rosales JE, de la Paz AM, Rattner BA, Rice CP (2003) Determination of alkylphenol and alkylphenolethoxylates in biota by liquid chromatography with detection by tandem mass spectrometry and fluorescence spectroscopy. J Chromatogr A 1010(1):25–35. CrossRefGoogle Scholar
  36. Schneider MJ, Lehotay SJ, Lightfield AR (2015) Validation of a streamlined multiclass, multiresidue method for determination of veterinary drug residues in bovine muscle by liquid chromatography-tandem mass spectrometry. Anal Bioanal Chem 407(15):4423–4435. CrossRefGoogle Scholar
  37. Stolker AAM, Brinkman UAT (2005) Analytical strategies for residue analysis of veterinary drugs and growth-promoting agents in food-producing animals—a review. J Chromatogr A 1067(1-2):15–53. CrossRefGoogle Scholar
  38. Turnipseed SB, Storey JM, Lohne JJ, Andersen WC, Burger R, Johnson AS, Madson MR (2016) Wide-scope screening method for multiclass veterinary drug residues in fish, shrimp, and eel using liquid chromatography–quadrupole high-resolution mass spectrometry. J Agric Food Chem 65:7252–7267. CrossRefGoogle Scholar
  39. Uchida K, Konishi Y, Harada K, Okihashi M, Yamaguchi T, Do MHN, Thi Bui L, Duc Nguyen T, Do Nguyen P, Thi Khong D, Thi Tran H, Nam Nguyen T, Viet le H, van Chau V, Thi van Dao K, Thi Ngoc Nguyen H, Kajimura K, Kumeda Y, Tran Pham K, Ngoc Pham K, Trong Bui C, Quang Vien M, Hoang le N, van Dang C, Hirata K, Yamamoto Y (2016) Monitoring of antibiotic residues in aquatic products in urban and rural areas of Vietnam. J Agric Food Chem 64(31):6133–6138. CrossRefGoogle Scholar
  40. USDA United States Department of Agriculture (2017) Maximum Residue Limits (MRL) Database. Retrieved from : Google Scholar
  41. Van Hoof N, Courtheyn D, Antignac J-P, Van de Wiele M, Poelmans S, Noppe H, De Brabander H (2005) Multi-residue liquid chromatography/tandem mass spectrometric analysis of beta-agonists in urine using molecular imprinted polymers. Rapid Comm Mass Spectrom 19(19):2801–2808. CrossRefGoogle Scholar
  42. Villar-Pulido M, Gilbert-López B, García-Reyes JF, Martos NR, Molina-Díaz A (2011) Multiclass detection and quantitation of antibiotics and veterinary drugs in shrimps by fast liquid chromatography time-of-flight mass spectrometry. Talanta 85(3):1419–1427. CrossRefGoogle Scholar
  43. WHO World Health Organization (2014) Antimicrobial resistance: global report on surveillance [Internet] Available from:

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Pesticide and Veterinary Drugs Residue DivisionNational Institute of Food and Drug Safety EvaluationCheongjuSouth Korea
  2. 2.Busan Regional Office, Ministry of Food and Drug SafetyBusanSouth Korea

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