RP-HPLC法同时测定酶法合成阿莫西林工艺中的原料、中间体和产物的含量
Simultaneous determination of content of raw materials,intermediates, and products in process of enzymatic synthesis of amoxicillin by RP-HPLC

目的:建立同时测定酶法合成阿莫西林反应体系中原料、中间体和产物含量的反相高效液相分析方法。方法:以 Gemini-NX-C18为固定相; 甲醇？？GA6FA？？0.05 mol/L、pH 5.6的磷酸盐缓冲液10？？GA6FA？？90(V？？GA6FA？？V)为流动相; 流速1.0 mL/min; 检测波长230 nm; 进样量10 μL; 柱温30 ℃进行检测。结果:6-氨基青霉烷酸、D-对羟基苯甘氨酸甲酯、D-对羟基苯甘氨酸、阿莫西林在各自的线性条件内线性关系良好,最低检测限分别为4.00、1.00、0.04、1.00 mg/L。结论:该方法准确灵敏,适合同时测定酶法合成阿莫西林反应体系中原料、中间体和产物的含量。
Aim: To establish a RP-HPLC method to simultaneously determine the content of raw materials, intermediates, and products in the process of enzymatic synthesis of amoxicillin.Methods: The determination was performed on a Gemini-NX-C18 column with a 10？？GA6FA？？90 mixture of methanol and phosphate buffer solution(0.05 mol/L, pH 5.6)as mobile phase, 1.0 mL/min as flow rate, 230 nm as determination wavelength, and 10 μL as injection volume with column temperature of 30 ℃.Results: The content of 6-APA, HPGM, HPG, and amoxicillin had good linear relationship in certain range and the limits of the detection were 4.00, 1.00, 0.04, and 1.00 mg/L, respectively.Conclusion: The method is accurate, sensitive and suitable to simultaneously determine the content of raw materials, intermediates, and products in the process of enzymatic synthesis of amoxicillin