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Enzyme immunoassay and liquid chromatography-fluorescence detection for amikacin in raw milk

Yiqiang Chen1, Qian Chen, Lidong He

  • 1State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.

Food Chemistry
|August 8, 2012
PubMed

An enzyme immunoassay and a liquid chromatography (LC) method for amikacin (AMK) in raw milk were developed in this study. Anti-AMK antibody was prepared by immunizing rabbits with AMK-BSA conjugate. The developed immunoassay exhibited an IC(50) value of 1.30 ng/mL and the spiked recoveries at 25-1000 ng/mL ranged from 69.8% to 93.3% with coefficients of variation (CVs) of 8.5-17.6%. For LC analysis, AMK was derivatized with 9-fluorenylmethyl chloroformate, which was followed by C8 column separation and fluorescence detection. By trichloroacetic acid extraction and MCX column purification, the recoveries at spiked concentrations of 500-5000 ng/mL were 80.7-91.3% with CVs less than 6.3%. The two methods can be selectively used for rapidly screening or quantitatively determining AMK in raw milk.

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