基于HPLC指纹图谱及多成分定量测定的痹祺胶囊质量评价

Quality evaluation of Biqi Capsules based on HPLC fingerprint and multicomponent analysis

目的建立痹祺胶囊的HPLC指纹图谱及甘草苷、阿魏酸、丹酚酸B的含量测定方法,更为全面地评价痹祺胶囊的质量。方法采用Shiseido Capcell Pak C_(18)MG II色谱柱(250 mm×4.6 mm,5μm),指纹图谱以乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,体积流量1.0 mL/min,检测波长203 nm,柱温为35℃,测定19批痹祺胶囊指纹图谱,结合中药色谱指纹图谱相似度评价系统进行全面分析;含量测定以乙腈-水-0.1%磷酸水溶液(21∶79∶0.1)为流动相;体积流量1.0 mL/min;以二极管阵列检测器检测,甘草苷检测波长为276 nm,阿魏酸检测波长为321 nm,丹酚酸B检测波长为286 nm;柱温为40℃;进样体积均为10μL。结果19批痹祺胶囊指纹图谱相似度大于0.97,共标定了36个共有峰,确认了6号峰为丹参素、10号峰为士的宁、11号峰为马钱子碱、13号峰为甘草苷、14号峰为阿魏酸、15号峰为迷迭香酸、16号峰为人参皂苷Rg1、18号峰为丹酚酸B、20号峰为人参皂苷Rb1、22号峰为甘草酸铵、27号峰为藁本内酯、30号峰为丹参酮IIA、33号峰为茯苓酸13个成分;含量测定中甘草苷、阿魏酸、丹酚酸B进样量在30.35~455.30 ng、5.046~75.690 ng、61.79~926.90ng线性关系良好(均为r=0.9999);平均回收率在98.43%~102.12%,RSD均<2%;19批痹祺胶囊中甘草苷质量分数在0.630~1.428 mg/g,阿魏酸质量分数在0.188~0.260 mg/g,丹酚酸B质量分数在2.539~3.442 mg/g。结论建立的指纹图谱及含量测定方法适用于痹祺胶囊的全面质量评价。

Objective To establish the HPLC fingerprint of Biqi Capsules(痹祺胶囊)and the determination method of the content of liquiritin,ferulic acid and salvianolic acid B,and evaluate the quality of Biqi Capsules more comprehensively.Methods Shiseido Capcell Pak C_(18) MG II chromatography column(250 mm×4.6 mm,5μm)was used,fingerprint mobile phase was acetonitrile-0.1%phosphate for gradient elution,the flow rate was 1.0 mL/min,the wavelength was 203 nm,the column temperature was 35℃,and the fingerprints of 19 batches of Biqi Capsules were measured.Combined with the Chinese medicine chromatography fingerprint similarity evaluation system,the Biqi Capsules was comprehensively analyzed;Content mobile phase was acetonitrile-water-0.1%phosphoric acid(21:79:0.1),the flow rate was 1.0 mL/min;With diode array detector,the detection wavelength of liquiritin was 276 nm,the detection wavelength of ferulic acid was 321 nm,and the detection wavelength of salvianolic acid B was 286 nm,the column temperature was 40℃,and the injection volume was 10μL.Results The similarity of 19 batches of Biqi Capsules was greater than 0.97.A total of 36 common peaks were calibrated,with peak 6 identified as danshensu,peak 10 as strychnine,peak 11 as brucine,peak 13 as liquiritin,peak 14 as ferulic acid,peak 15 as rosmarinic acid,peak 16 as ginsenoside Rg1,peak 18 as salvianolic acid B,peak 20 as ginsenoside Rb1,peak 22 as ammonium glycyrrhizinate,peak 27 as ligustilide,peak 30 as tanshinone IIA,and peak 33 as pachymic acid;The inlet quantity of liquiritin,ferulic acid and salvianolic acid B have good linear relationships in the range of 30.35—455.30 ng,5.046—75.690 ng,61.79—926.90 ng,respectively(r=0.9999).The average recovery rates were 98.43%—102.12%,and RSD<2%,liquiritin content was 0.630—1.428 mg/g,ferulic acid content was 0.188—0.260 mg/g,and salvianolic acid B content was 2.539—3.442 mg/g.Conclusion The established fingerprint and content determination method are suitable for the comprehensive quality evaluation of Biqi Capsules.