Abstract: A high performance capillary electrophoresis (HPCE) method was developed for the simultaneous determination of penicillin intermediate and penicillins in milk, including 6-amino-penicillanic acid (6-APA), penicillin G (PEN), ampicillin (AMP) and amoxicillin (AMO). The main parameters including the ion concentration and pH value of running buffer, separation voltage and column temperature were optimized systematically by orthogonal test. The four penicillins (PENs) were baseline separated within 4.5 min with the running buffer of 40 mmol/L potassium dihydrogen phosphate-20 mmol/L borax solution (pH 7.8), separation voltage of 28 kV and column temperature of 30 ℃. The calibration curves showed good linearity in the range of 1.56~100 mg/L, and the correlation coefficients (r2) were between 0.9979 and 0.9998. The average recoveries at three spiked levels were in the range of 84.91%~96.72% with acceptable relative standard deviations (RSDs) of 1.11%~9.11%. The method is simple, fast, accurate and suitable for the determination of penicillins in real samples.

Key words: 6-aminopenicillanic acid, milk, penicillin intermediate, penicillins, high performance capillary electrophoresis(HPCE)