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Development and validation of an HPLC method for the determination of seven penicillin antibiotics in veterinary drugs and bovine blood plasma

Samanidou, Victoria F., Giannakis, Dimitrios E., Papadaki, Antonia
Journal of separation science 2009 v.32 no.9 pp. 1302-1311
adenosine monophosphate, amoxicillin, blood plasma, cattle, colchicine, detection limit, dicloxacillin, freeze-thaw cycles, high performance liquid chromatography, penicillin V, veterinary drugs
Herein a quantitative method for the determination of seven penicillins in bovine plasma and veterinary drugs has been developed. Amoxicillin (AMO), ampicillin (AMP), penicillin G (PENG), penicillin V (PENV), oxacillin (OXA), cloxacillin (CLO) and dicloxacillin (DICLO) were separated on a Perfectsil ODS-2 (250x4 mm, 5 μm) column, using gradient elution, with a mobile phase of 0.1% v/v TFA and ACN-methanol (90:10 v/v). PDA detection was used at 240 nm. Penicillins were isolated from bovine plasma by SPE on Lichrolut RP-18 cartridges with mean recoveries from 85.7 to 113.5%. Colchicine (3 ng/μL) was used as an internal standard. The developed method was validated in terms of selectivity, linearity, accuracy, precision, stability and sensitivity. Repeatability (n = 5) and between-day precision (n = 5) revealed RSD < 12%. The detection limits in the bovine plasma were estimated as 18 ng for AMO and AMP, 25 for PENG, PENV and OXA, 3 ng for CLO and 12 ng for DICLO. Spiked plasma samples were stable for 1 wk, except for AMP and CLO, which were stable for 3 wk and OXA for 4 wk. AMO, PENG and PENV were stable for two freeze-thaw cycles, OXA, CLO and DICLO for four, while AMP only for one.