双丹颗粒HPLC指纹图谱研究及7个成分的含量测定

Study on the HPLC Fingerprint and Content Determination of Seven Constituents in Shuangdan Granules

目的:建立双丹颗粒(丹参、牡丹皮)HPLC指纹图谱,并测定其中7个成分的含量。方法:采用Agilent TC-C_(18)(250 mm×4.6 mm,5μm)色谱柱;流动相为乙腈-0.05%磷酸水溶液,梯度洗脱;流速1.0 ml·min^(-1);检测波长220 nm;柱温:30℃。采用《中药色谱指纹图谱相似度评价系统(2012版)》建立15批双丹颗粒的HPLC指纹图谱,并进行相似度评价,与混合对照品色图谱比对指认共有峰,并测定其含量。结果:15批样品指纹图谱中有26个共有峰,与对照指纹图谱的相似度均不低于0.950。指认化学成分7个,分别为没食子酸、丹参素、芍药苷、迷迭香酸、丹酚酸B、丹皮酚、丹参酮Ⅱ_(A),且在各自范围内线性关系良好(r≥0.9950),平均加样回收率为99.4%-102.8%,RSD为0.31%-1.38%,15批样品中上述成分的含量分别为0.6120-0.9940 mg·g^(-1),2.1060-2.5000 mg·g^(-1),0.4360-0.6470 mg·g^(-1),0.8750-1.6670 mg·g^(-1),3.7960-4.7410 mg·g^(-1),2.0010-2.9650 mg·g^(-1),1.0020-1.4210 mg·g^(-1)。结论:本研究建立了双丹颗粒的HPLC指纹图谱和含量测定方法,可用于双丹颗粒的质量控制。

Objective:To establish the HPLC fingerprint of Shuangdan granules(Salviae miltiorrhizae Radix et Rhizoma,Moutan cortex),and to determine the contents of 7 components.Methods:An HPLC method was adopted.The separation was performed on an Agilent TC-C_(18) column(250 mm×4.6 mm,5μm)with the mobile phase consisting of acetonitrile and 0.05%phosphoric acid flowing at 1.0 ml·min^(-1) in a gradient elution manner,the detection wavelength was set at 220 nm,and the column temperature was set at 30℃.HPLC fingerprints of 15 batches of Shuangdan granule were established by using Similarity Evaluation System of TCM Chromatographic Fingerprint(2012 edition),the common peaks were identified by comparing with the mixed reference substance,and the contents were determined as well.Results:There were 26 common peaks in the fingerprints of 15 batches of samples,and the similarities with the control fingerprint were not less than 0.950.Seven components were identified,which were gallic acid,danshensu,paeoniflorin,rosmarinic acid,salvianolic acid B,paeonol and tanshinoneⅡ_(A),and all showed good linear relationships within their own ranges(r≥0.9950),whose average recoveries were 99.4%-102.8%with the RSDs of 0.31%-1.38%.In 15 batches of samples,the contents of the above 7 components were 0.6120-0.9940 mg·g^(-1),2.1060-2.5000 mg·g^(-1),0.4360-0.6470 mg·g^(-1),0.8750-1.6670 mg·g^(-1),3.7960-4.7410 mg·g^(-1),2.0010-2.9650 mg·g^(-1) and 1.0020-1.4210 mg·g^(-1),respectively.Conclusion:In this study,HPLC fingerprint and content determination method for Shuangdan granules are successfully established,which can be used for the quality control.