目的:建立HPLC法同时测定丹参-当归药对中丹参素、绿原酸、阿魏酸、迷迭香酸、丹酚酸B、藁本内酯和丹参酮ⅡA 7个成分含量,明确配伍对7个成分含量变化的影响。方法:应用RP-HPLC多波长切换梯度洗脱技术,采用Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5 μm),以乙腈-0.1%磷酸水溶液为流动相,流速1.0 mL·min-1,检测波长280 nm(丹参素、丹参酮ⅡA)、286 nm(丹酚酸B)和320 nm(绿原酸、阿魏酸、迷迭香酸、藁本内酯),柱温25℃。结果:丹参素、绿原酸、阿魏酸、迷迭香酸、丹酚酸B、藁本内酯和丹参酮ⅡA质量浓度分别在7.263~145.3、4.002~200.1、6.433~160.8、10.57~528.3、209.0~1.045×104、111.7~5 584、6.027~301.3 ng·mL-1范围内与峰面积呈良好的线性关系(r=0.999 5~1.000),平均加样回收率在97.1%~101.9%之间(RSD<3%,n=6)。丹参-当归配伍后丹参素和藁本内酯的含量显著增加(P<0.05),而绿原酸、阿魏酸、迷迭香酸、丹酚酸b和丹参酮ⅱa含量无明显变化。3个批次的丹参、当归饮片中上述7个成分含量测定结果分别为0.237 7~0.264 1、0.384 8~0.455 5、0.687 3~0.955 7、1.018~1.768、39.32~54.64、10.65~14.98、1.675~2.608 mg·g-1。结论:该方法可用于丹参-当归药对及其制剂的质量控制,并为丹参、当归的临床配伍应用及其研究提供科学依据。
Objective:To establish an HPLC method for simultaneous determination of danshensu, chlorogenic acid, ferulic acid, rosmarinic acid, salvianolic acid B, ligustilide and tanshinoneⅡA in Radix Salviae Miltiorrhizae-Radix Angelicae Sinensis drug pair, so as to analyse the variation of the 7 ingredients after compatibility. Methods:The samples were separated on an Eclipse XDB-C18 column(4.6 mm×250 mm, 5 μm) by gradient elution with acetonitrile-0.1% phosphoric acid aqueous solution at a flow rate of 1.0 mL·min-1. The detection wavelength was 280 nm (danshensu and tanshinoneⅡA), 286 nm (salvianolic acid B) and 320 nm (chlorogenic acid, ferulic acid, rosmarinic acid and ligustilide). The column temperature was controlled at 25. Results:Danshensu, chlorogenic acid, ferulic acid, rosmarinic acid, salvianolic acid B, ligustilide and tanshinoneⅡA exhibited good linearity (r=0.999 5-1.000) among the ranges of 7.263-145.3, 4.002-200.1, 6.433-160.8, 10.57-528.3, 209.0-1.045×104, 111.7-5 584 and 6.027-301.3 ng·mL-1, respectively. The average recoveries (RSD<3%, n=6) were between 97.1%-101.9%. After the compatibility, the content of danshensu and ligustilide increased significantly (P<0.05), while there was no significant change in the content of chlorogenic acid, ferulic acid, rosmarinic acid, salvianolic acid B and tanshinoneⅡA. The contents of the above-mentioned compounds in 3 batches of samples were 0.237 7-0.264 1, 0.384 8-0.455 5, 0.687 3-0.955 7, 1.018-1.768, 39.32-54.64, 10.65-14.98 and 1.675-2.608 mg·g-1, respectively. Conclusion:The method can be used for the quality control of Salviae Miltiorrhizae Radix and Angelicae Sinensis Radix drug pair and its preparations, and can provide scientific basis for the clinical compatibility and application of Salviae Miltiorrhizae Radix and Angelicae Sinensis Radix.