HPLC法测定丹参-红花配方颗粒中5种成分的含量

Content Determination of 5 Constituents in Danshen-Honghua Dispensing Granule by HPLC

目的:建立高效液相色谱法测定丹参-红花配方颗粒中丹酚酸B、丹参素、紫草酸、迷迭香酸、羟基红花黄色素A的含量。方法:采用ThermoHypersilGOLDC18色谱柱(250 mm×4.6 mm,5μm);流动相为乙腈-0.2%磷酸水溶液;梯度洗脱;柱温:25℃;检测波长:288 nm;流速:1.0 mL/min。结果:丹酚酸B、丹参素、紫草酸、迷迭香酸、羟基红花黄色素A分别在31.50~315.00 ng(r=0.9995)、8.40~84.00 ng(r=0.9991)、3.60~36.00 ng(r=0.9994)、5.80~58.00 ng(r=0.9991)、6.70~67.00 ng(r=0.9990)范围内线性关系良好,各成分的平均加样回收率范围在94.51%~99.09%,RSD均低于4.9%。3批样品中丹酚酸B、丹参素、紫草酸、迷迭香酸、羟基红花黄色素A的含量分别为37.27~42.21 mg/g、7.11~8.11 mg/g、4.48~4.94 mg/g、2.74~3.50 mg/g、和1.39~1.76 mg/g。结论:该方法简便、准确、快速,重复性好、稳定性好,可用于丹参-红花配方颗粒中5种成分的含量测定。

Objective:To establish a high performance liquid chromatography(HPLC)method for the content determination of salvianolic acid B,danshensu,lithospermic acid,rosmarinic acid,and hy⁃droxysafflor yellow A in the Danshen-Honghua dispensing granule.Methods:The separation was per⁃formed on the Thermo Hypersil GOLD C18 column(250 mm×4.6 mm,5μm)with acetonitrile-0.2%phosphoric acidas the mobile phase by gradient elution at flow rate of 1.0 mL/min.The column tempera⁃ture was 25℃,and the detection wavelength was 288 nm.Results:salvianolic acid B,danshensu,litho⁃spermic acid,rosmarinic acid,and hydroxysafflor yellow A were successfully separated with this meth⁃od,with good linearity at 31.5-315.00 ng(r=0.9995),8.40-84.00 ng(r=0.9991),3.60-36.00 ng(r=0.9994),5.80-58.00 ng(r=0.9991),6.70-67.00 ng(r=0.9990),respectively.The range of their average recovery was 94.51%-99.09%,and the RSD of results were less than 4.9%.The con⁃tents of salvianolic acid B,danshensu,lithospermic acid,rosmarinic acid,and hydroxysafflor yellow A in three batches of samples were 37.27-42.21 mg/g,7.11-8.11 mg/g,4.48-4.94 mg/g,2.74-3.50 mg/g and 1.39-1.76 mg/g.Conclusion:The established method is simple,accurate,rapid,reproduc⁃ible,well stability,which are suitable for the determination of 5 constituents in the Danshen-Honghua dispensing granule.