紫参片的HPLC指纹图谱研究及5个成分含量测定

目的: 研究建立紫参片的HPLC法指纹图谱分析方法,并测定该制剂中藁苯内酯、正丁烯基苯酞、丹参酮Ⅰ、隐丹参酮、丹参酮Ⅱ_A的含量,为其质量控制提供依据。 方法: 利用HPLC法,采用Ultimate^® XB C₁₈色谱柱(250 mm×4.6 mm,5 μm),使用乙腈-0.3%磷酸水溶液为流动相进行梯度洗脱,以260 nm为检测波长。在紫参片样品的HPLC图中,以丹参酮Ⅱ_A为对照参比物,对各色谱峰与丹参酮Ⅱ_A色谱峰的相对保留时间和相对峰面积的RSD进行分析,运用中国药典委员会"中药色谱指纹图谱相似度评价系统2004A版"对10批紫参片的图谱进行相似度评价。采用外标法,利用藁苯内酯、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_A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32个峰构成紫参片的指纹特征,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;各批次样品中丹参酮Ⅱ_A色谱峰所对应的保留时间一致;每批样品的各色谱峰与丹参酮Ⅱ_A色谱峰的相对保留时间和相对峰面积的RSD均符合指纹图谱的检测要求,10个批次样品的指纹图谱相似度均大于0.940,5个物质在10批该制剂中的含量基本一致。 结论: HPLC指纹图谱方法与5个物质的含量测定方法准确、稳定、简便,均可用于紫参片的质量控制。

Objective: To establish an HPLC method for fingerprint analysis of Zishen tablets,and determination of the contents such as ligustilide,n-butylene(n-butene) phthalide,cryptotanshinone,tanshinone Ⅰ,and tanshinoneⅡ_A,so as to provide a scientific basis for the quality control. Methods: The high performance liquid chromatography(HPLC) method was adopted,and Kromasil KR100-5C₁₈(250 mm×4.6 mm,5 μm)column was used,acetonitrile-0.3% phosphoric acid solution was selected as mobile phase adopting gradient elution,and determination wavelength was 260 nm.In the recorded chromatogram of the test preparation,tanshinoneⅡ_A was used as 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 Zishen tablets was appraised by the similarity evaluation system.Using the external standard method and mixture reference substance ,their contents were determined in the 10 batches of preparation. Results: In this chromatogram condition,32 peaks were identified as the characteristic fingerprints of Zishen tablets.Ten batches of Zishen tablet samples showed the same character.The retention time for tanshinone Ⅱ_A were consistent with each other.RSD(%)of the relative retention time and the relative peak areas of all peaks compared with tanshinoneⅡ_A fit the requirements of fingerprint.The fingerprints showed good similarity up to 94% in 10 batches of Zishen tablets.Five compounds' contents were almost the same in 10 batches of preparation. Conclusion: The method is accurate,stable and simple,which can be used for quality control of Zishen tablets.