小儿清热止咳口服液高效液相色谱指纹图谱的建立及8种成分含量测定

Establishment of HPLC Fingerprint and Determination of Eight Components in Xiaoer Qingre Zhike Oral Liquid

目的建立小儿清热止咳口服液的高效液相色谱(HPLC)指纹图谱,并测定其中8种主要成分的含量。方法色谱柱为Agilent Infinity Lab Poroshell 120 EC-C18柱(150 mm×4.6 mm,2.7μm),流动相为乙腈-甲醇(89∶11,V/V)-0.01 mol/L磷酸二氢钾溶液(pH 3.2),梯度洗脱,流速为1.0 mL/min,检测波长为210 nm(0~31 min)、270 nm(31~75 min),柱温为30℃,进样量为10μL。结果共得到28个共有峰,其中26个可找到归属,各批样品间相似度均大于0.99。盐酸麻黄碱、黄芩苷、苦杏仁苷、甘草酸铵、盐酸伪麻黄碱、黄芩素、甘草苷、(R,S)-告依春质量浓度分别在2.05~41.06μg/mL、38.42~768.48μg/mL、9.00~179.93μg/mL、3.23~64.61μg/mL、1.09~21.90μg/mL、1.49~29.80μg/mL、1.94~38.87μg/mL、0.69~13.71μg/mL范围内与峰面积线性关系良好(r≥0.9994),检测限为0.0686~0.8576μg/mL,定量限为0.3284~1.6152μg/mL;稳定性、重复性、中间精密度试验结果的RSD均小于3.0%;平均加样回收率为97.90%~101.44%,RSD为0.45%~1.85%(n=6)。结论该方法灵敏、简便、准确,可用于小儿清热止咳口服液的质量控制。

Objective To establish a high-performance liquid chromatography(HPLC)fingerprint of Xiaoer Qingre Zhike Oral Liquid,and to determine the contents of eight main components.Methods The chromatographic column was Agilent Infinity Lab Poroshell 120 EC-C18 column(150 mm×4.6 mm,2.7μm),the mobile phase was acetonitrile-methanol(89∶11,V/V)-0.01 mol/L potassium dihydrogen phosphate solution(pH 3.2),with gradient elution,the flow rate was 1.0 mL/min,the detection wavelengths were 210 nm(0-31 min)and 270 nm(31-75 min),the column temperature was 30℃,and the injection volume was 10μL.Results A total of 28 common peaks were obtained,of which 26 could be attributed specific medicinal herb,and the similarity of different batches of samples was greater than 0.99.The linear ranges of ephedrine hydrochloride,baicalin,amygdalin,ammonium glycyrrhizinate,pseudoephedrine hydrochloride,baicalein,liquiritin and(R,S)-epigoitrin were 2.05-41.06μg/mL,38.42-768.48μg/mL,9.00-179.93μg/mL,3.23-64.61μg/mL,1.09-21.90μg/mL,1.49-29.80μg/mL,1.94-38.87μg/mL,0.69-13.71μg/mL(r≥0.9994)respectively.The limit of detection(LOD)and the limit of quantification(LOQ)were in the range of 0.0686 to 0.8576μg/mL,0.3284 to 1.6152μg/mL respectively.The RSDs of stability,repeatability and intermediate precision tests were all lower than 3.0%.The average recovery rate of the above eight components was in the range of 97.90%to 101.44%,with the RSD in the range of 0.45%to 1.85%(n=6).Conclusion This method is sensitive,simple and accurate,which can be used for the quality control of Xiaoer Qingre Zhike Oral Liquid.