蓝芩颗粒高效液相色谱指纹图谱的建立及9个指标成分含量测定

Establishment of HPLC Fingerprint of Lanqin Granules and Content Determination of Nine Indicator Components

目的建立蓝芩颗粒的高效液相色谱(HPLC)指纹图谱,并测定其中指标成分含量。方法色谱柱为Agilent Infinity LabPoroshell 120 EC-C_(18)柱(150 mm×4.6 mm,2.7μm),流动相为乙腈-0.02 mol/L磷酸二氢钠溶液(p H 4.2)、梯度洗脱,流速为1.0 m L/min,检测波长为230 nm,柱温为30℃,进样量为10μL。采用中药色谱指纹图谱相似度评价系统(2012版)绘制12批样品的特征图谱,计算相似度;确定共有峰并指认,并同时测定指认出的成分含量。结果蓝芩颗粒HPLC指纹图谱中有23个共有峰,相似度均大于0.92。指认出其中9个,分别为(R,S)-告依春、京尼平龙胆双糖苷、栀子苷、盐酸黄柏碱、黄芩苷、千层纸素A-7-O-β-D-葡萄糖醛酸苷、汉黄芩苷、盐酸小檗碱、汉黄芩素,其质量浓度分别在0.0262~7.815μg/m L、0.2040~411.600μg/m L、0.1222~377.400μg/m L、0.1951~15.360μg/m L、0.0787~297.000μg/m L、0.0920~410.103μg/m L、0.2019~149.625μg/m L、0.1018~7.538μg/m L、0.1019~19.200μg/m L范围内与峰面积线性关系良好(r≥0.9998);检测限为0.0065~0.0511μg/m L,定量限为0.0262~0.2040μg/m L;中间精密度、稳定性、重复性试验结果的RSD均小于3.0%,平均加样回收率为96.37%~101.69%,RSD为0.48%~0.91%(n=6)。结论该方法灵敏、简便、准确,可用于蓝芩颗粒的质量控制。

Objective To establish a high-performance liquid chromatography(HPLC)fingerprint of Lanqin Granules,and to determine the contents of nine indicator components.Methods The chromatographic column was the Agilent Infinity Lab Poroshell 120 EC-C_(18)column(150 mm×4.6 mm,2.7μm),the mobile phase was acetonitrile-0.02 mol/L sodium dihydrogen phosphate solution(pH 4.2)with gradient elution,the flow rate was 1.0 mL/min,the detection wavelength was 230 nm,the column temperature was 30℃,and the injection volume was 10μL.The characteristic HPLC fingerprint of 12 batches of samples was established by the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Version),and the similarity was calculated;the common peaks were marked and identified,and the contents of identified components were determined.Results There were 23 common peaks in the HPLC fingerprint of Lanqin Granules,with a similarity greater than 0.92.Nine of them were identified,including(R,S)-goitrin,genipin 1-gentiobioside,geniposide,phellodendrine chloride,baicalin,oroxyloside,wogonoside,berberine hydrochloride,wogonin.The contents of the above nine components were 0.0262-7.815μg/mL,0.2040-411.600μg/mL,0.1222-377.400μg/mL,0.1951-15.360μg/mL,0.0787-297.000μg/mL,0.0920-410.103μg/mL,0.2019-149.625μg/mL,0.1018-7.538μg/mL,0.1019-19.200μg/mL(r≥0.9998),respectively.The limits of detection(LOD)were in the range of 0.0065-0.0511μg/mL,and the limits of quantification(LOQ)were in the range of 0.0262-0.2040μg/mL.The RSDs of the intermediate precision,stability and repeatability tests were all lower than 3.0%.The average recovery rates of the above nine components were in the range of 96.37%-101.69%,with the RSDs in the range of 0.48%-0.91%(n=6).Conclusion This method is sensitive,simple,and accurate,which can be used for the quality control of Lanqin Granules.