Determination of the Persistence of Tetracycline Antibiotics and Their Degradates in Manure-Amended Soil Using Enzyme-Linked Immunosorbent Assay and Liquid Chromatography−Mass Spectrometry

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Abstract
The persistence of manure-borne oxytetracycline in soil was investigated under field conditions. Soil cores were collected approximately once a month for over a period of two years and subsampled at depth increments of 0−5, 5−10, 10−15, 15−36, and 36−71 cm. Soil samples were analyzed by enzyme-linked immunosorbent assay (ELISA) and/or by liquid chromatography−mass spectrometry (LC-MS). Whereas LC-MS showed that oxytetracycline declined to <50% of its initial soil concentration after 3 weeks, ELISA showed that the total tetracyclines did not decline significantly 5 months after manure application. The differences between ELISA and LC-MS results are attributed to the broad cross-reactivity of the antibodies employed, which detect many structurally related tetracyclines, including their isomers and degradation products. Only trace amounts (≤1.0 μg/kg) of oxytetracycline were observed in the subsurface soil, and none was detected in water samples from field lysimeters, suggesting that oxytetracycline has low mobility in soil. Keywords: Antibiotics; manure; ELISA; LC-MS; persistence; mobility

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