Determination of Multiclass Harmful Residues in Shellfish Using LC-MS

Xiaoming Gong, Ronghui Ma, Kai Li, Hongwei Zhang, Zhenxing Wu, Hongtao Wang, Jun Sun, Han Zhao


A multi-residue procedure with liquid chromatography-tandem mass spectrometry analysis was developed to simultaneously detect the presence of 4 diarrheic shellfish poisoning toxins, 14 sulfonamides, chloramphenicol and thiamphenicol in shellfish tissues. Samples were first purified by a modified rapid procedure and separated on a ZOBRAX Eclipse Plus C18 column and quantified using electron spray ionization-mass spectrometry-mass spectrometry. The results showed that the limits of quantification (LOQ) of SPX1, OA-C, GYM, and PTX-2 were 1.0, 2, 0.5 and 2 μg/kg, respectively. The LOQ for the antibiotics was below acceptable limits established by most countries. Compared to the external standard method, this matrix matched the calibration method effectively, overcame the matrix effects and gave better quantitative results. Recoveries of spiked compounds ranged between 67.6% and 109.8%, with relative standard deviations below 15% for most target analytes. Only sulfathiazole had a %RSD of 18.6% at the lowest spiked concentration. The proposed method was accurate, rapid and reliable. (DOI assigned 4/17/2019)


LC-MS/MS, diarrheic shellfish poisoning toxins, multiclass residues, matrix effects

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Amending Annex III to Regulation (EC) No 853/2004 of the European Parliament and of the Council as regards the permitted limits of yessotoxins in live bivalve molluscs Text with EEA relevance, (EU) No 786/2013 (2013).

Bittencourt, M. S., Martins, M. T., de Albuquerque, F. G. S., Barreto, F., & Hoff, R. (2012). High-throughput multiclass screening method for antibiotic residue analysis in meat using liquid chromatography-tandem mass spectrometry: a novel minimum sample preparation procedure. Food Additives and Contaminants Part a-Chemistry Analysis Control Exposure & Risk Assessment, 29(4), 508-516.

Fusetani, N., & Kem, W. (2009). Marine toxins: an overview. Progress in Molecular and Subcellular Biology, 46, 1-44.

Fux, E., McMillan, D., Bire, R., & Hess, P. (2007). Development of an ultra-performance liquid chromatography-mass spectrometry method for the detection of lipophilic marine toxins. Journal of Chromatogrophy A, 1157(1-2), 273-280.

Gerssen, A., Mulder, P. P., McElhinney, M. A., & de Boer, J. (2009). Liquid chromatography-tandem mass spectrometry method for the detection of marine lipophilic toxins under alkaline conditions. Journal of Chromatogrophy A, 1216(9), 1421-1430.

James, K. J., Lehane, M., Moroney, C., Fernandez-Puente, P., Satake, M., Yasumoto, T., & Furey, A. (2002). Azaspiracid shellfish poisoning: unusual toxin dynamics in shellfish and the increased risk of acute human intoxications. Food Additives and Contaminants, 19(6), 555-561.

Laying Down Specific Hygiene Rules for Food of Animal Origin, (EC) No 853/2004 (2004).

Lehotay, S. J. (2011). QuEChERS sample preparation approach for mass spectrometric analysis of pesticide residues in foods. Methods in Molecular Biology, 747, 65-91.

Pharmacologically Active Substances and Their Classification Regarding Maximum Residue Limits in Foodstuffs of Animal Origin, (EU) No 37/2010 (2009).

Quilliam, M. A. (2003). The role of chromatography in the hunt for red tide toxins. Journal of Chromatography A, 1000(1-2), 527-548.

Suzuki, H. (2013). Differences in Susceptibility to Okadaic Acid, a Diarrhetic Shellfish Poisoning Toxin, between Male and Female Mice. Toxins, 5(1), 9-15.

Suzuki, T., & Quilliam, M. A. (2011). LC-MS/MS Analysis of Diarrhetic Shellfish Poisoning (DSP) Toxins, Okadaic Acid and Dinophysistoxin Analogues, and Other Lipophilic Toxins. Analytical Sciences, 27(6), 571-584.

Tian, W. L., Gao, L. Y., Zhao, Y. Z., Peng, W. J., & Chen, Z. Z. (2013). Simultaneous determination of metronidazole, chloramphenicol and 10 sulfonamide residues in honey by LC-MS/MS. Analytical Methods, 5(5), 1283-1288.

Yasumoto, T., Oshima, Y., & Yamaguchi, M. (1978). Occurrence of a New Type of Shellfish Poisoning in the Tohoku District. Nippon Suisan Gakkaishi, 44(11), 1249-1255.