Abstract
A rapid, simple, reliable, and sensitive immunogold chromatographic assay (IGCA) was described for simultaneous determination of four macrolide antibiotics (erythromycin, spiramycin, tilmicosin, and tylosin) in raw milk. Three antigens were immobilized as three test lines on the nitrocellulose membrane, which enable the simultaneous determination on a single test strip. Samples were detected directly without treatment; the entire testing process was completed within 10 min. The visual detection limits for erythromycin, spiramycin, tilmicosin, and tylosin were 5, 5, 10, and 20 ng/mL, respectively. Sixty blind raw milk samples were analyzed by both IGCA and liquid chromatography–tandem mass spectrometry; the results showed a good correlation between the two methods. The results demonstrate that the developed method could provide a rapid and effective approach for the onsite determination of multi-macrolide antibiotics residues in a large number of samples.
Similar content being viewed by others
References
Albrecht U, Hammer P, Heeschen W (1996) Chicken antibody based ELISA for the detection of spiramycin in raw milk. Milchwissenschaft-Milk Sci Int 51:209–212
Albrecht U, Walte HG, Hammer P (1998) Detection of erythromycin in raw milk by an antibody-capture-immunoassay. Kiel Milchwirtschaftliche Forschungsberichte 50:163–170
Berrada H, Borrull F, Font G, Marce RM (2008) Determination of macrolide antibiotics in meat and fish using pressurized liquid extraction and liquid chromatography-mass spectrometry. J Chromatogr A 1208:83–89
Billedeau SM, Heinze TM, Siitonen PH (2003) Liquid chromatography analysis of erythromycin A in salmon tissue by electrochemical detection with confirmation by electrospray ionization mass spectrometry. J Agric Food Chem 51:1534–1538
Breier AR, Garcia CV, Oppe TP, Steppe M, Schapoval EES (2002) Microbiological assay for azithromycin in pharmaceutical formulations. J Pharm Biomed Anal 29:957–961
Chen Y, Wang Z, Tang S, Zhu Y, Xiao X (2008) Rapid enzyme-linked immunosorbent assay and colloidal gold immunoassay for kanamycin and tobramycin in swine tissues. J Agric Food Chem 56:2944–2952
Deubel A, Fandino AS, Soergel F, Holzgrabe U (2006) Determination of erythromycin and related substances in commercial samples using liquid chromatography/ion trap mass spectrometry. J Chromatogr A 1136:39–47
Heinrich K, Macarthur R, von Holst C, Sharman M (2013) An inter-laboratory validation of a multiplex dipstick assay for four classes of antibiotics in honey. Anal Bioanal Chem 24:7875–7884
Horie M, Saito K, Ishii R, Yoshida T, Haramaki Y, Nakazawa H (1998) Simultaneous determination of five macrolide antibiotics in meat by high-performance liquid chromatography. J Chromatogr A 812:295–302
Kibwage IO, Roets E, Hoogmartens J (1983) Thin-layer chromatography of erythromycins and other macrolides. J Chromatogr 256:164–171
Le T, He HQ, Niu XD, Chen Y, Xu J (2013) Development of an immunochromatographic assay for detection of tylosin and tilmicosin in muscle, liver, fish and eggs. Food Agric Immunol 4:467–480
Li XM, Luo PJ, Tang SS, Beier RC, Wu XP, Yang LL, Li YW, Xiao XL (2011) Development of an immunochromatographic strip test for rapid detection of melamine in raw milk, milk products and animal feed. J Agric Food Chem 59:6064–6070
Lucchetti D, Fabrizi L, Esposito A, Guandalini E, Di Pasquale M, Coni E (2005) Simple confirmatory method for the determination of erythromycin residues in trout: a fast liquid-liquid extraction followed by liquid chromatography-tandem mass spectrometry. J Agric Food Chem 53:9689–9694
Luo XQ, Wan YP, Sun Z, Wang SL, Tian T, Cui TT (2012) Determination of tylosin and tilmicosin in milk by gold immune chromatographic assay. China Dairy Ind 40:42–45
Maher HM, Youssef RM, Khalil RH, El-Bahr SM (2008) Simultaneous multiresidue determination of metronidazole and spiramycin in fish muscle using high performance liquid chromatography with UV detection. J Chromatogr B 876:175–181
Paek SH, Lee SH, Cho JH (2000) Development of rapid one-step immunochromatographic assay. Methods 22:53–60
Peng DP, Ye SQ, Wang YL, Chen DM, Tao YF, Huang LL, Liu ZL, Dai MH, Wang XQ, Yuan ZH (2012) Development and validation of an indirect competitive enzyme-linked immunosorbent assay for the screening of tylosin and tilmicosin in muscle, liver, milk, honey and eggs. J Agric Food Chem 60:44–51
The Maximum residue limits of Veterinary drugs in Animal Food. Announcement No. 235 (2002) of the Ministry of Agriculture of the People’s Republic of China
Wang J, Leung D, Lenz SP (2006) Determination of five macrolide antibiotic residues in raw milk using liquid chromatography-electrospray ionization tandem mass spectrometry. J Agric Food Chem 54:2873–2880
Zhang JK, Qi YH, Liu JX, Wang JP (2013) Heterologous immunoassay for screening macrolide antibiotics residues in milk based on the monoclonal antibody of tylosin. Food Agric Immunol 24:419–431
Acknowledgments
This work is supported financially by an National Key Technology R&D Program from the Ministry of Science and Technology (No. 2012BAK17B16), and the Trans-Century Training Programme Foundation for Talents by the Ministry of Education (NCET-12–0529) and the State Key Program of the National Natural Science of China (No. 30830082).
Conflict of Interest
Xiangmei Li has no competing financial interests. Kai Wen has no competing financial interests. Yiqiang Chen has no competing financial interests. Xiaoping Wu has no competing financial interests. Xingyao Pei has no competing financial interests. Qi Wang has no competing financial interests. Ailing Liu has no competing financial interests. Jianzhong Shen has no competing financial interests. Guidelines for the care and use of laboratory animals were followed.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Li, X., Wen, K., Chen, Y. et al. Multiplex Immunogold Chromatographic Assay for Simultaneous Determination of Macrolide Antibiotics in Raw Milk. Food Anal. Methods 8, 2368–2375 (2015). https://doi.org/10.1007/s12161-015-0130-x
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12161-015-0130-x