口服液体制剂中8种抑菌剂的HPLC检测与抑菌效力评价  

Determination by HPLC and Evaluation of Antimicrobial Effectiveness of 8 Antibacterial Agents of Oral Liquid

作　　者：蔡金莲 郭智聪 黄婉锋 Cai Jinlian;Guo Zhicong;Huang Wanfeng(Foshan Center for Food and Drug Control,Foshan 528000,China)

机构地区：[1]佛山市食品药品检验检测中心,广东佛山528000

出　　处：《广东化工》2023年第16期173-177,共5页Guangdong Chemical Industry

基　　金：2020年佛山市自筹经费类科技计划项目(医学类科技攻关项目,2020001005459)。

摘　　要：目的:建立口服液体制剂中8种抑菌剂的HPLC检测方法,对20批样品进行抑菌剂检测并进行抑菌效力检查和评价。方法:色谱柱:Agilent C18柱(4.6×250 mm,5μm),以流动相20 mmol/L乙酸铵(A)-甲醇(B)进行梯度洗脱,检测波长:230 nm、254 nm,柱温30℃,流速1 m L/min。按2020年版《中国药典(四部)》通则1121抑菌效力检查法,以金黄色葡萄球菌、铜绿假单胞菌、大肠埃希菌、白色念珠菌和黑曲霉为试验菌株,分别加至20批口服液体制剂中,进行抑菌效力试验,检测微生物在不同时间点的存活菌落数。结果:8种抑菌剂浓度在0.2~20μg/mL范围内与峰面积线性关系良好,线性相关系数R2≥0.9991,检出限(S/N=3)为0.007~0.1μg/mL,加标平均回收率为99.4%~100.2%(RSD为0.1%~0.5%)。20批样品中,有2批样品羟苯甲酯不符合药典的规定,有11批样品的抑菌效力不符合药典的规定。结论:8种抑菌剂的HPLC检测方法简单可靠,可用于口服液体制剂中抑菌剂的监测,口服液体制剂中抑菌剂的添加有待进一步的规范。Objective:To establish an HPLC method for simultaneous determination of 8 antibacterial agents,determine the same 8 antibacterial agents with the method and evaluate the antimicrobial effectiveness of 20 batches of oral liquid.Methods:The chromatographic separation was achieved on a Agilent C18(4.6 mm×150 mm,5μm)column with 20 mmol/L ammonium acetate as mobile phase A and methanol as mobile phase B(gradient elution).The detection wavelength was 230 nm and 254 nm,column temperature was 30℃,flow rate was 1 mL/min.According to the requirements of antimicrobial effectiveness of the general rule 1121 in the Chinese Pharmacopoeia(2020 edition,volumeⅣ),Staphylococcus aureus,Pseudomonas aeruginosa,Escherichia coli,Candida albicans and Aspergillus niger were used as the test strains and added to the 20 batches of oral liquid for the challenge test of microorganism,the number of viable bacteria in samples was determined at different time points.Results:The linear range of the method of 8 antibacterial agents was 0.2~20μg/mL(R2≥0.9991),detection limit(S/N=3)was 0.007~0.1μg/mL,the average recoveries was 99.4%~100.2%(RSD was 0.1%~0.5%).The content of methyl hydroxybenzoate in 2 batches was exceeding the standard of Chinese Pharmacopoeia,and the antimicrobial effectiveness of 11 samples failed to meet the requirements of the Chinese Pharmacopoeia.Conclusion:The method is simple and reliable and can be used as daily monitoring method for the antibacterial agents.The use of antibacterial agents needs further standardization.

关 键 词：口服液体制剂 HPLC 抑菌剂 抑菌效力评价 存活菌落数 

分 类 号：TQ[化学工程]