杏仁止咳合剂HPLC指纹图谱及7种成分的含量测定

Study on HPLC Fingerprints and Content Determination of 7 Components in Xingren Zhike Mixture

目的建立杏仁止咳合剂HPLC指纹图谱,同时测定7种成分含量。方法采用HPLC-DAD法,Agilent ZORBAX Eclipse Plus C18色谱柱(250 mm×4.6 mm,5μm);流动相:乙腈(A)-0.1%磷酸水溶液(B),梯度洗脱;流速为1.0 mL·min^-1;检测波长:210 nm(苦杏仁苷、细叶远志皂苷)、251 nm(甘草酸铵)、237 nm(甘草苷)、284 nm(橙皮苷、柚皮苷、新橙皮苷);柱温30℃;进样量10μL。利用中药色谱指纹图谱相似度评价系统进行相似度评价,采用SPSS 22.0统计软件对含量测定结果进行聚类分析。结果15批样品指纹图谱有20个共有峰,相似度>0.90。苦杏仁苷、细叶远志皂苷、甘草苷、柚皮苷、橙皮苷、甘草酸铵、新橙皮苷检测质量浓度线性范围分别为31.97~1278μg·mL^-1,26.40~1056μg·mL^-1,40.38~1615μg·mL^-1,2.252~90.10μg·mL^-1,7.698~307.9μg·mL^-1,3.742~149.7μg·mL^-1,3.032~121.3μg·mL^-1(r≥0.9991);定量限≤1.901μg·mL^-1;精密度、稳定性(48 h)、重复性试验的RSD<2.0%(n=6或n=7);平均加样回收率分别为99.1%,96.2%,101.1%,96.9%,97.4%,98.1%,99.2%(n=9);15批样品聚为3类。结论本法简便、准确、重复性好,可以用于杏仁止咳合剂的鉴定与质量评价,可作为杏仁止咳合剂质量控制的重要科学依据。

OBJECTIVE To establish the HPLC fingerprint of Xingren Zhike mixture and determine the contents of 7 chemical compositions.METHODS HPLC-DAD analysis was performed on Agilent ZORBAX Eclipse Plus C18(250 mm×4.6 mm,5μm)with the column temperature of 30℃and the injection volumn was 10μL.The mobile phase was acetonitrile(A)-0.1%phosphoric acid solution(B)with gradient elution at the flow rate of 1.0 mL·min^-1.The detection wavelength was 210 nm for amygdalin and tenuifolin;237 nm for liquiritin;251 nm for ammonium glycyrrhizate;284 nm for hesperidin,naringin and neohesperidin.The similarity evaluation system for chromatographic fingerprint of TCM was used to calculate the similarity and the cluster analysis was conducted for the results of content determination by SPSS 22.0 statistical software.RESULTS There were 20 common peaks in the HPLC fingerprints of 15 batches of samples with the similarities>0.90.The linear ranges were 31.97-1278μg·mL^-1 for amygdalin,26.40^-1056μg·mL^-1 for tenuifolin,40.38-1615μg·m L^-1 for liquiritin,2.252-90.10μg·mL^-1 for naringin,7.698-307.9μg·mL^-1 for hesperidin,3.742-149.7μg·mL^-1 for ammonium glycyrrhizate,3.032-121.3μg·mL^-1 for neohesperidin.The limits of quantitation were≤1.901μg·mL^-1,with r≥0.9991.RSDs of precision,reproducibility and stability tests(48 h)were≤2.0%(n=6 or n=7).Average recoveries were 99.1%,96.2%,101.1%,96.9%,97.4%,98.1%,99.2%(n=9),respectively.The samples of 15 batches of Xingren Zhike mixture could be divided into 3 categories.CONCLUSION The method is simple and accurate with good repeatability,which can be used for the identification and evaluation of Xingren Zhike mixture,and provides important scientific basis for the quality control of Xingren Zhike mixture.