Colistin, an imperative member of the polymyxin group, is a cationic peptide antibiotic. Itis also known as polymyxin E, but this peptide antibiotic has been forbidden for human consumption due to its high toxicity. Regrettably, this antibiotic is utilized as a feed additive and veterinary drug for animals. Due to the toxicity of colistin, the presence of its residue in the animal system represents a threat to human health regarding the consumption of meat, especially chicken. A novel method was proposed for quantifying colistin B in chicken muscles and eggs using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). In this method, extraction of colistin B from samples was achieved by mixing the sample with acidified methanol:water (1/1, v/v), followed by centrifugation and filtration by a membrane filter excluding solid-phase extraction (SPE) clean up, as well as evaporation steps. The analysis was conducted by optimized liquid chromatography-tandem mass spectrometry (LC-MS/MS), and method performance was assessed in terms of the limit of quantitation, specificity, selectivity, precision, linearity and recovery in coherence with the guidelines of SANTE and the Commission Decision 2002/657/EC. The result obtained from the study showed the limit of quantitation (LOQ) as 10 µg Kg-1 for muscles and 5 µg Kg-1 for eggs, with acceptable recoveries along with precision. The linearity was plotted in the range of 5-25 µg L-1 (solvent) for egg and 10-50 µg Kg-1 (matrix-matched) for muscles. The result of average recoveries showed the value of 70-94% (3.3-12% relative standard deviation (RSD)) for chicken muscles and 88-107% (2.5-18.6% RSD) for egg samples, which meets the criteria for acceptability of method according to both SANTE and 2002/657/EC guidelines. This proposed protocol provides a cost-effective solution for food testing labs by reducing the cost of the sample preparation by 60% along with the time required for SPE cleanup. Further, the optimized method was also tested on real samples collected from nearby provinces in Solan city, Himachal Pradesh, India, and three out of 20 muscles were found to have colistin B in the range of 50-560 µg Kg-1.

Department of Chemistry Faculty of Science University of Hradec Kralove 50003 Hradec Kralove Czech Republic

Food Safety Solution Center FSSAI Thermo Fisher Scientific Indirapuram Ghaziabad 201014 India

School of Bioengineering and Food Technology Shoolini University of Biotechnology and Management Sciences Solan 173229 India

School of Biological and Environmental Sciences Shoolini University of Biotechnology and Management Sciences Solan 173229 India

School of Pharmaceutical Sciences Shoolini University of Biotechnology and Management Sciences Solan 173229 India

Zobrazit více v PubMed

Kumar H., Chen B.H., Kuca K., Nepovimova E., Kaushal A., Nagraik R., Bhatia S.K., Dhanjal D.S., Kumar V., Kumar A., et al. Understanding of colistin usage in food animals and available detection techniques: A review. Animals. 2020;10:1892. doi: 10.3390/ani10101892. PubMed DOI PMC

Bladek T., Szymanek B.I., Posyniak A. Determination of polypeptide antibiotic residues in food of animal origin by ultra-high-performance liquid chromatography-tandem mass spectrometry. Molecules. 2020;25:3261. doi: 10.3390/molecules25143261. PubMed DOI PMC

Catry B., Cavaleri M., Baptiste K., Grave K., Grein K., Holm A., Jukes H., Liebana E., Lopez Navas A., Mackay D., et al. Use of colistin-containing products within the European Union and European Economic Area (EU/EEA): Development of resistance in animals and possible impact on human and animal health. Int. J. Antimicrob. Agents. 2015;46:297–306. doi: 10.1016/j.ijantimicag.2015.06.005. PubMed DOI

Liu Y.Y., Wang Y., Walsh T.R., Yi L.X., Zhang R., Spencer J., Doi Y., Tian G., Dong B., Huang X., et al. Emergence of plasmid-mediated colistin resistance mechanism mcr-1 in animals and human beings in China: A microbiological and molecular biological study. Lancet Infect. Dis. 2016;16:161–168. doi: 10.1016/S1473-3099(15)00424-7. PubMed DOI

Davies M., Walsh T.R. A colistin crisis in India. Lancet Infect. Dis. 2018;18:256–257. doi: 10.1016/S1473-3099(18)30072-0. PubMed DOI

Livemint Govt May Ban Antibiotic Colistin Used to Fatten Chicken. [(accessed on 20 June 2019)]; Available online: https://www.livemint.com/Industry/yt5eE5hqMLYP1px2d63Q1K/Govt-may-ban-antibiotic-colistin-used-to-fatten-chicken.html.

TOI Tolerance Limits’ to Be Fixed by Food Regulator for Presence of Antibiotics in Animal, Foods. [(accessed on 12 June 2019)]; Available online: https://www.fssai.gov.in/upload/media/FSSAI_News_AntiBiotics_TOI_01_08_2018.pdf.

FSSAI Direction under Section 16(5)Read with 18(2)(d)of Food Safety and Standard Act, 2006 Regarding Operationalisation of Draft Food Safety and Standard (Contaminants, Toxins and Residues) Amendment Regulations. [(accessed on 8 September 2019)];2019 Available online: https://www.fssai.gov.in/upload/advisories/2019/08/5d4c042779d77Direction_Colistin_Ban_FSSAI_08_08_2019.pdf.

Sin D.W., Ho C., Wong Y.C., Ho S.K., Ip A.C.B. Analysis of major components of residual bacitracin and colistin in food samples by liquid chromatography tandem mass spectrometry. Anal. Chim. Acta. 2005;535:23–31. doi: 10.1016/j.aca.2004.11.063. DOI

Wan E.C., Ho C., Sin D.W., Wong Y.C. Detection of residual bacitracin A, colistin A, and colistin B in milk and animal tissues by liquid chromatography tandem mass spectrometry. Anal. Bioanal. Chem. 2006;385:181–188. doi: 10.1007/s00216-006-0325-5. PubMed DOI

Xu I., Tian X., Ren C., Huang H., Zhang X., Gong X., Liu H., Yu Z., Zhang L. Analysis of colistin A and B in fishery products by ultra performance liquid chromatography with positive electro spray ionization tandem mass spectrometry. J. Chromatogr. B. 2012;899:14–20. doi: 10.1016/j.jchromb.2012.04.028. PubMed DOI

Boison J.O., Lee S., Matus J. A multi-residue method for the determination of seven polypeptide drug residues in chicken muscle tissues by LC-MS/MS. Anal. Bioanal. Chem. 2015;406:4065–4078. doi: 10.1007/s00216-015-8644-z. PubMed DOI

Pascale R., Bianco G., Coviello D., Lafiosca M.C., Masi S., Mancini I.M., Bufo S.A., Scrano L., Caniani D. Validation of a LC-MS/MS method for the determination of drugs in wastewater using a three-phase solvent system. J. Sep. Sci. 2020;43:886–895. doi: 10.1002/jssc.201900509. PubMed DOI

SANTE. [(accessed on 8 December 2020)]; Available online: https://ec.europa.eu/food/sites/food/files/plant/docs/pesticides_mrl_guidelines_wrkdoc_2019-12682.pdf.

European Communities Commission Decision (2002/657/EC) of 12 August 2002 implementing Council Directive 96/23/EC concerning the performance of analytical methods and the interpretation of results. Off. J. Eur. Communities. 2002;L221:8–36.

Fu Q., Li X., Zheng K., Ke Y., Wang Y., Wang L., Yu F., Xia X. Determination of colistin in animal tissues, egg, milk, and feed by ultra-high performance liquid chromatography-tandem mass spectrometry. Food Chem. 2018;248:166–172. doi: 10.1016/j.foodchem.2017.12.029. PubMed DOI

Saluti G., Diamanti I., Giusepponi D., Pucciarini L., Rossi R., Moretti S., Sardella R., Galarini R. Simultaneous determination of aminoglycosides and colistins in food. Food Chem. 2018;266:9–16. doi: 10.1016/j.foodchem.2018.05.113. PubMed DOI

QuPPe-Method. [(accessed on 10 December 2020)]; Available online: https://www.eurl-pesticides.eu/userfiles/file/EurlSRM/meth_QuPPe_AO.pdf.

Electrochemical immunosensor for the detection of colistin in chicken liver