采用高效液相色谱法建立了同时检测牛奶中青霉素类抗生素中间体6-氨基青霉烷酸(6-APA)及2种青霉素类药物氨苄青霉素(AMP)、阿莫西林(AMO)的方法。检测条件如下:色谱柱为Xterra?MS C18柱;流动相为乙腈:0.1%氨水=5:95(V/V)等度洗脱;柱温35℃;流速1.0 mL·min-1;进样量10μL;紫外检测波长200 nm。结果表明,6 min内可实现3种青霉素类抗生素(PENs)的快速分离检测。方法在0.125~10.00 mg·L-1范围内有良好的线性,相关系数(r2)0.9972~0.9991,加标回收率为89.09%~99.64%,相对标准偏差为1.21%~7.35%。该方法简便、快速,可应用于市售牛奶中PENs的快速检测。 A method for the simultaneous determination of 6-aminopenicillanic acid (6-APA) and 2 penicillins ampicillin (AMP) and amoxicillin (AMO) in penicillin antibiotics was established by high performance liquid chromatography. The detection conditions were as follows: the column was Xterra MS 18 column; the mobile phase was acetonitrile: 0.1% aqueous ammonia = 5:95 (V / V); the column temperature was 35 ℃; the flow rate was 1.0 mL · min -1; Amount of 10μL; UV detection wavelength of 200 nm. The results showed that 3 kinds of penicillin antibiotics (PENs) could be rapidly separated and detected within 6 min. The method has a good linearity in the range of 0.125-10.00 mg · L-1, the correlation coefficient (r2) is 0.9972-0.9991, the spiked recoveries are 89.09% -99.64%, and the relative standard deviations are 1.21% -7.35%. The method is simple, rapid and can be applied to the rapid detection of PENs in commercial milk.