基于色谱法筛查及验证杞菊地黄口服液中抑菌剂

Screening and Verification of Bacteriostatic Agents in Qiju Dihuang Oral Liquid by Chromatography

目的 筛查并验证杞菊地黄口服液中的抑菌剂。方法 采用高效液相色谱(HPLC)法测定苯甲酸、山梨酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、对羟基苯甲酸丁酯的含量,色谱柱为Welch Ultimate Plus C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-0.02 mol/L乙酸铵溶液(梯度洗脱),流速为1.0 mL/min,检测波长为255 nm,柱温为35℃,进样量为20μL;采用液相色谱串联质谱(LC-MS/MS)法验证制剂中的对羟基苯甲酸乙酯。结果 苯甲酸、山梨酸、对羟基苯甲酸甲酯、对羟基苯甲酸乙酯、对羟基苯甲酸丙酯、对羟基苯甲酸丁酯的进样量分别在0.065 97~3.298 5μg、0.051 71~2.585 5μg、0.034 36~1.718 0μg、0.015 18~0.759 0μg、0.031 67~1.583 5μg、0.035 48~1.774 0μg范围内与峰面积响应值线性关系良好(r≥0.999 9,n=7);精密度、稳定性、重复性试验结果的RSD均小于2.0%;加样回收率分别为102.61%,103.00%,99.53%,95.95%,100.10%,100.45%,RSD分别为0.90%,0.50%,0.57%,0.14%,0.39%,0.43%(n=6)。12批样品违规使用对羟基苯甲酸乙酯,5批样品未检出药品说明书中规定的抑菌剂。结论 所建立的方法科学、简便、专属性较强,可用于口服液中常见6种抑菌剂的快速筛查。杞菊地黄口服液中抑菌剂的使用种类及用量存在与标示不符的情况。

Objective To screen and verify the antibacterial agents in Qiju Dihuang Oral Liquid.Methods High-performance liquid chromatography(HPLC) method was used to determine the contents of benzoic acid,sorbic acid,methyl 4-hydroxybenzoate,ethyl 4-hydroxybenzoate,propyl 4-hydroxybenzoate,butyl 4-hydroxybenzoate,the chromatographic column was Welch Ultimate Plus C18column(250 mm × 4.6 mm,5 μm),the mobile phase was acetonitrile-0.02 mol/L ammonium acetate solution(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 255 nm,the column temperature was 35 ℃,and the injection volume was 20 μL.The liquid chromatography tandem-mass spectrometry(LC-MS/MS) was used to verify the ethylparaben in the preparation.ResultsThe linear ranges of benzoic acid,sorbic acid,methyl 4-hydroxybenzoate,ethyl 4-hydroxybenzoate,propyl 4-hydroxybenzoate,butyl 4-hydroxybenzoate were 0.065 97-3.298 5 μg,0.051 71-2.585 5 μg,0.034 36-1.718 0 μg,0.015 18-0.759 0 μg,0.031 67-1.583 5 μg,0.035 48-1.774 0 μg(r ≥ 0.999 9,n = 7).The RSDs of precision,stability,and repeatability test results were all lower than 2.0%.The recovery rates of benzoic acid,sorbic acid,methyl 4-hydroxybenzoate,ethyl 4-hydroxybenzoate,propyl 4-hydroxybenzoate,butyl 4-hydroxybenzoate were 102.61%,103.00%,99.53%,95.95%,100.10%,and 100.45%,with RSDs of 0.90%,0.50%,0.57%,0.14%,0.39%,and 0.43%(n = 6),respectively.Illegal use of ethylparaben in the samples was confirmed in 12 batches of samples.No bacteriostatic agents lablelled in the drug manual were dectected in five batches of samples.Conclusion The established method is scientific,simple,and highly specific,which can be used for rapid screening of six common bacteriostatic agents in oral fluid.There is a discrepancy between the varieties and amounts of bacteriostatic agents used in Qiju Dihuang Oral Liquid and the labeling.