Microbiological and thin-layer chromatographic identification of benzylpenicillin and ampicillin in animal body.

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Abstract
Benzylpenicillin (PG-G) and ampicillin (AB-PC) were administered i.v. to pigs and calves. An attempt was made to identify these penicillins qualitatively in tissues and urine collected from the animals after slaughter. The kidney, liver and muscle were homogenized into 4-fold dilutions. The diameter of an inhibition zone produced by each dilution was determined on Micrococcus luteus ATCC 9341 and Bacillus subtilis ATCC 6633 plates at pH 6.0 and 8.0. The urine was diluted up to 625-fold. TLC identification was performed for only tissue and urine samples characterized by the largest inhibition zone that was observed on M. luteus plate and inactivated by the addition of penicillinase. Next, a sample extracted with methanol was spotted on a silica gel plate, which was then developed in ethyl acetate-methanol-water (6:2:1). Rf values were determined by bioautography with M. luteus as the test organism. They were about 2 times as high in animals given PC-G as in those given AB-PC. They coincided closely with those of reference antibiotic solutions. Thus, it seems likely that PC-G and AB-PC residues may be identified in slaughtered animals by TLC with the spot volume of 10 and/or 30 .mu.l on the basis of inhibition zones produced on M. luteus plate. It was recommended that chromatography should be performed on extracts from the medulla rather than cortex of the kidney, which had been proved to contain penicillins.

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