高效液相色谱法测定β-内酰胺抗生素中残留乙酸的含量  

Determination of residual acetic acid in the raw materials of β-lactam antibiotics using high performance liquid chromatography

作　　者：冯艳春[1] 裴文莉 王晨[1] 朱俐[1] 田冶[1] 崇小萌[1] 宋暤昀 姚尚辰[1] 宁保明[1] Feng Yanchun;Pei Wenli;Wang Chen;Zhu Li;Tian Ye;Chong Xiaomeng;Song Haoyun;Yao Shangchen;Ning Baoming(National Institutes for Food and Drug Control,Beijing 102629)

机构地区：[1]中国食品药品检定研究院,北京102629

出　　处：《中国抗生素杂志》2024年第4期388-393,共6页Chinese Journal of Antibiotics

摘　　要：目的建立测定β-内酰胺类抗生素原料中乙酸残留量的HPLC方法。方法采用多因素组内设计方法,确定色谱流动相的组成、比例和pH值。参考ICH Q2,从专属性、检出限和定量限、线性、准确度、精密度和耐用性等6个方面对方法进行验证。用所建立的方法对19种β-内酰胺类抗生素原料进行了测定。结果采用C_(18)色谱柱,流动相:甲醇-0.02 mol/L磷酸二氢钾,梯度洗脱;柱温25℃;检测波长220 nm。供试品、溶剂和流动相对乙酸测定没有明显干扰;乙酸的最低检出限约为0.029μg,最低定量限约0.12μg;从定量限到限度2倍浓度范围内,方法的线性关系良好,拟合直线方程为y=388866.7186x-177.5720,r2=0.9999;低中高浓度加样回收率平均分别为93%,96%和99%;方法的重复性和中间精密度分别为2.37%和2.11%;在3根不同C_(18)色谱柱上的耐用性良好。19种β-内酰胺抗生素原料中有3种筛查出残留乙酸。结论所建方法可以实现对β-内酰胺类抗生素原料中残留乙酸的准确定量,为该类原料的生产过程控制和标准物质研制提供了依据。Objective To establish high performance liquid chromatography(HPLC)for the determination of residual acetic acid inβ-lactam raw materials.Methods A multifactorial within-group design method was used to determine the composition,ratio,and pH of the chromatographic mobile phase.The established method was validated in terms of specificity,limit of detection,limit of quantification,linearity,accuracy,precision,and robustness with reference to ICH Q2.This developed method identified 19β-lactam antibiotic raw materials.Results The separation was carried out on a C_(18)column with the mobile phase of methanol-0.02 mol/L potassium dihydrogen phosphate solution in gradient elution;the column temperature was 25℃,and the detection wavelength was 220 nm.There was no obvious interference with the determination of acetic acid by the test samples,solvent,or mobile phase.The lowest detection limit of acetic acid was about 0.029μg and the lowest limit of quantification was about 0.12μg.The linearity of the method was good in the range of concentration from the limit of quantification to 2 times the limit concentration of acetic acid.And the fitted linear equation was y=388866.7186x-177.5720,r2=0.9999.The average recoveries of the spiked samples at low,medium,and high concentrations were 93%,96%and 99%respectively;the repeatability and intermediate precision of the method were 2.37%and 2.11%respectively.The robustness of the method was good on the three different kinds of C_(18)columns.Residual acetic acid was tested in three out of the 19β-lactam antibiotics raw materials.Conclusion The proposed method could achieve accurate quantification of residual acetic acid inβ-lactam raw materials,which provided a reference for process control and the content assignment of standard substances for such raw materials.

关 键 词：Β-内酰胺类抗生素 原料 乙酸 反相高效液相色谱法 

分 类 号：R978.1[医药卫生—药品] R917[医药卫生—药学]