SIMULTANEOUS IDENTIFICATION AND DETERMINATION OF RESIDUAL PENICILLINSBY USE OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH SPECTROPHOTOMETRIC OR FLUOROMETRIC DETECTORS
Cc. Hong et al., SIMULTANEOUS IDENTIFICATION AND DETERMINATION OF RESIDUAL PENICILLINSBY USE OF HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY WITH SPECTROPHOTOMETRIC OR FLUOROMETRIC DETECTORS, American journal of veterinary research, 56(3), 1995, pp. 297-303
Using 7 penicillins (amoxicillin, ampicillin, methicillin, penicillin
G, oxacillin, cloxacillin, and dicloxacillin), simultaneous and direct
determination of residual penicillins in biological samples was carri
ed out by use of bioassay and high-performance liquid chromatography w
ith spectrophotometric or fluorometric detectors. By use of assay medi
um seeded with penicillin-sensitive Micrococcus luteus (ATCC No. 9341)
as a test organism, we were able to detect penicillins even at low co
ncentrations. All penicillins treated with 10 U of penicillinase/ml di
d not produce inhibition zones by disk testing, even at a concentratio
n of 100 mu g of penicillin/ml/assay plate. Using a mobile phase of ac
etonitrile:methanol:0.01M KH2PO4 (19:11:70, v/v/v; pH, 7.1), standard
solutions of the penicillins were separated from each other by use of
high-performance liquid chromatography analysis, producing symmetric p
eaks without tailing, each of which had a characteristic retention tim
e. Simultaneous detection of residual penicillins in bovine serum kidn
eys, and liver, for the 5 penicillins for which analysis was possible
by use of the UV method, yielded recovery rates from 71.4 to 102.3%; f
or the 2 amino-penicillins, amoxicillin and ampicillin, which could on
ly be detected by use of the fluorometric method, recovery rate ranged
from 72.9 to 103%.