丹参多酚酸对照提取物的质量控制

目的:建立丹参多酚酸对照提取物多指标含量测定及指纹图谱的质量控制方法。方法:采用Agilent ZORBAX SB-C18色谱柱(4.6 mm×250 mm,5μm),以0.1%甲酸-水溶液为流动相A,0.1%甲酸-乙腈溶液为流动相B,梯度洗脱(0~30 min,20%~21.5%B;30~35 min,21.5%~25%B;35~45 min,25%~40%B;45~50 min,40%~95%B),柱温30℃,流速1 mL·min^-1,检测波长288 nm;采用浓度法测定咖啡酸,丹酚酸E,迷迭香酸,紫草酸,丹酚酸B,丹酚酸Y的相对校正因子,并测定丹参多酚酸对照提取物各指标成分含量,与单体对照品外标法测定结果进行比较;同时建立指纹图谱方法,并对10批次提取物进行相似度评价。结果:咖啡酸,丹酚酸E,迷迭香酸,紫草酸,丹酚酸B,丹酚酸Y在各自的检测质量浓度范围内线性关系良好(r>0.9999),进样精密度RSD在0.1%~1.2%,重复性RSD在1.2%~1.6%,6种成分的加样回收率在82.03%~98.68%,样品溶液在36 h内各成分稳定性良好。经测定各指标成分的相对校正因子分别为咖啡酸(2.92),丹酚酸E(1.10),迷迭香酸(1.61),紫草酸(1.07),丹酚酸B(1.00),丹酚酸Y(0.83)。考察不同方法、浓度、仪器、色谱柱、波长等因素的影响,发现测得的相对校正因子适用性较好。校正因子法测定结果和单体对照品外标法测定结果差异较小。建立了丹参多酚酸对照提取物的HPLC特征指纹图谱,确定了5个共有特征峰,并根据对照品对色谱峰进行确认,10批次提取物指纹图谱相似度较高,质量差异较小。结论:该研究建立的多指标含量测定方法和指纹图谱方法简便、快速、准确性高、重复性好,可用于丹参多酚酸对照提取物的质量控制。

Quality Control of Salvianolic Acids Reference Extract

Objective:To establish the quality control method for multi-index content determination and fingerprint of salvianolic acids.Method:Agilent ZORBAX SB-C18(4.6 mm×250 mm,5μm)column was adopted,with 0.1%formic acid-water as mobile phase A and 0.1%formic acid-acetonitrile as mobile phase B for gradient elution(0-30 min,20%-21.5%B;30-35 min,21.5%-25%B;35-45 min,25%-40%B;45-50 min,40%-95%B).The column temperature was set at 30℃,the flow rate was set at 1 mL·min^-1,and the detection wavelength was set at 288 nm.Relative correction factors of caffeic acid,salvianolic acid E,rosmarinic acid,lithosperic acid,salvianolic acid B and salvianolic acid Y were determined by the concentration method.The content of each indicator component of the reference extract of salvianolic acid polyphenolic acid was determined and compared with the results of the monomer reference substance by the external standard method.At the same time,the fingerprint method was established.and the similarity evaluation was carried out on 10 batches of extracts.Result:Caffeic acid,salvianolic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B,and salvianolic acid Y had a good linear relationship within the respective detection mass concentration ranges(r>0.9999).The injection precision RSD was 0.1%-1.2%,the reproducible RSD was 1.2%-1.6%,and the recovery of the six components was 82.03%-98.68%.The stability of each component in the sample solution was good within 36 h.The relative correction factors for each indicator component were determined to be caffeic acid(2.92),salvianolic acid E(1.10),rosmarinic acid(1.61),lithosperic acid(1.07),salvianolic acid B(1.00),salvianolic acid Y(0.83).The effects of different methods,concentrations,instruments,columns,wavelengths were investigated,and the measured relative correction factors were found to be suitable.The results of the calibration factor method and the monomer standard reference substance method were less different.The HPLC fingerprints of the reference extract of salvianolic acids were established,and five common characteristic peaks were determined.The chromatographic peaks were confirmed according to the reference substance.The similarity of the fingerprints of the 10 batches of extracts was higher,and the quality difference was smaller.Conclusion:The multi-index content determination method and the fingerprint method established in this study are simple,rapid,accurate and reproducible,and can be used for quality control of Salviae miltiorrhizae Radix et Rhizoma polyphenolic acid reference extract.