参花胶囊的HPLC指纹图谱研究及4种成分含量测定

目的研究建立参花胶囊的高效液相色谱(HPLC)指纹图谱分析方法,并测定该制剂中的绿原酸、羟基红花黄色素A、迷迭香酸、丹酚酸B的含量,为其质量控制提供依据。方法用HPLC法,采用Kromasil C18色谱柱(250mm×4.6mm,5μm);用乙腈-5.0g·L-1磷酸水溶液为流动相进行梯度洗脱;以254nm为检测波长。在参花胶囊样品的HPLC图中,以迷迭香酸为对照参比物,对各共有色谱峰与迷迭香酸色谱峰的相对保留时间与相对峰面积比值的RSD值进行分析,用中药色谱指纹图谱对10批参花胶囊进行相似度评价。采用外标法,以320nm为检测波长,运用绿原酸、羟基红花黄色素A、迷迭香酸、丹酚酸B的混合对照品测定其在10批该制剂中的含量。结果在选定的条件下确定39个峰构成参花胶囊的指纹特征,各批次样品均具有上述特征,且特征峰的相对含量分布基本一致;各批次样品中迷迭香酸色谱峰所对应的保留时间一致;每批样品的各色谱峰与迷迭香酸色谱峰的相对峰面积和相对保留时间的比值RSD均符合中药指纹图谱的相关要求,10个批次样品的指纹图谱相似度均大于0.966,4种物质在10批该制剂中的含量基本一致。结论 HPLC指纹图谱方法与4种物质的含量测定方法准确、稳定、简便,均可用于参花胶囊的质量控制。

Study on HPLC fingerprint of Shenhua Capsules and determining the four components

Objective To establish HPLC fingerprint of Shenhua Capsules ,to determine the content of chlorogenic acid ,hydroxysaf-flor yellow A ,rosmarinic acid ,salvianolic acid B ,and to provide the basis for its quality control .Methods A high performance liquid chromatography （HPLC） method was used ,and Kromasil C18 （250 mm × 4 .6 mm ,5 μm） column was used ,using acetonitrile-5 .0 g ？ L - 1 phosphoric acid solution as mobile phase in a gradient elution mode ,and determination wavelength was 254 nm .In the recorded chromatogram of the test preparation ,rosmarinic acid was used as the reference substance ,and RSD of the relative retention time and the relative peak areas of all peaks compared with its peak were measured .The similarity of 10 batches of Shen-hua Capsules was appraised by the similarity evaluation system .By using the external standard method ,and determination wave-length of 320 nm ,the contents of chlorogenic acid ,hydroxysafflor yellow A ,rosmarinic acid ,ralvianolic acid B were determined in 10 batches of the preparation .Results Under the chromatographic conditions ,39 peaks were identified as the characteristic fin-gerprint of Shenhua Capsules .10 batches of Shenhua Capsules samples showed the same characteristics .The retention time of ros-marinic acid was consistent with each other .RSD of the relative retention time and the relative peak areas of all peaks compared with rosmarinic acid fit the requirements of fingerprint .The fingerprints showed good similarity up to 0 .966 in 10 batches of Shen-hua Capsules .Four compounds′s contents were almost the same in 10 batches of the preparation .Conclusion The method is exact ,simple and accurate ,and can be used for the quality control of Shenhua Capsules .