蚁参蠲痹胶囊中5个成分测定及HPLC指纹图谱建立

Determination of five constituents Yishen Juanbi capsules and establishment of HPLC fingerprints

目的:测定蚁参蠲痹胶囊中桂皮醛、隐丹参酮、丹参酮Ⅰ、苍术素、丹参酮Ⅱ_(A)5个成分的含量,并建立蚁参蠲痹胶囊HPLC指纹图谱。方法:以丹参酮Ⅱ_(A)为内参物,建立桂皮醛、隐丹参酮、丹参酮Ⅰ和苍术素的相对校正因子,并进行含量计算,实现一测多评(QAMS),同时采用外标法测定3批蚁参蠲痹胶囊中上述5个化学成分的含量,比较计算值与实测值的差异,验证QAMS法的准确性;色谱条件:采用Kromasil 100-5 C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.02%磷酸水为流动相,梯度洗脱,流速1.0 mL·min^(-1),检测波长270 nm,柱温20℃。采用HPLC法,建立蚁参蠲痹胶囊HPLC指纹图谱,对共有峰进行归属,通过相似度评价、聚类分析、主成分分析对指纹图谱进行评价;色谱条件:采用Agilent 5 TC-C_(18)色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.02%磷酸水为流动相,梯度洗脱,流速1.0 mL·min^(-1),检测波长320、270 nm,柱温20℃。结果:建立的相对校正因子重现性良好,采用校正因子计算的含量值与实测值之间无显著差异,相对误差均在±2%以内。建立了蚁参蠲痹胶囊HPLC指纹图谱,共标定了36个共有峰,通过与对照品比较,指认出12个成分,10批样品指纹图谱与对照指纹图谱的相似度均大于0.95。采用QAMS法测得3批样品中桂皮醛、隐丹参酮、丹参酮Ⅰ、苍术素和丹参酮Ⅱ_(A)的含量分别为0.077~0.244、0.402~0.472、0.098~0.136、0.086~0.186、0.454~0.517 mg·g^(-1),QAMS法计算值与ESM实测值间均无显著性差异。结论:所建立的QAMS含量测定方法及HPLC指纹图谱简便、准确,重复性好,可为蚁参蠲痹胶囊的质量评价提供参考。

Objective:To determine the contents of five constituents(cinnamaldehyde,cryptotanshinone,tanshinoneⅠ,atractylodin,tanshinoneⅡ_(A))in Yishen Juanbi capsules and to establish the HPLC fingerprints.Methods:A quantitative analysis of multi-components by single-marker(QAMS)method was developed to determine cinnamaldehyde,cryptotanshinone,tanshinone I,atractylodin and tanshinoneⅡ_(A).TanshinoneⅡ_(A)was selected as internal reference and the relative correction factors(RCFs)of other four components were calculated.The QAMS method was evaluated by external standard method.The analysis was performed on a column of Kromasil 100-5 C_(18)(250 mm×4.6 mm,5μm)with gradient elution of acetonitrile-0.02%phosphoric acid aqueous solution at a flow rate of 1.0 mL·min^(-1).The detection wavelengths were set at 270 nm,and the column temperature was 20℃.The fingerprints of Yishen Juanbi capsules were established by HPLC,and evaluated by similarity evaluation,cluster analysis(CA)and principal component analysis(PCA).The analysis was performed on a column of Agilent 5 TC-C_(18)(250 mm×4.6 mm,5μm)with gradient elution of acetonitrile-0.02%phosphoric acid aqueous solution at a flow rate of 1.0 mL·min^(-1).The detection wavelengths were set at 320 nm and 270 nm,and the column temperature was 20℃.Results:The established RCFs had a good reproducibility.No significant differences were found between the quantitative results of external standard method and QAMS method.Relative errors of the two methods were within±2%.The HPLC fingerprint of Yishen Juanbi capsules was established with 36 common peaks,and 12 constituents were identified by comparing with the control product spectrogram.The similarities of 10 batches of samples were all above 0.95.The contents of cinnamaldehyde,cryptotanshinone,tanshinoneⅠ,atractylodin,tanshinoneⅡ_(A)in 3 batches of samples were 0.077-0.244,0.402-0.472,0.098-0.136,0.086-0.186,0.454-0.517 mg·g^(-1)respectively by QAMS.There was no significant difference between the QAMS and the ESM.Conclusion:The established method of QAMS and HPLC fingerprint analysis is simple,accurate and reproducible,and can provide a reference for the quality evaluation of Yishen Juanbi capsules.