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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.