HPLC波长切换法同时测定丹参酚酸提取物中5种成分

HPLC simultaneous determination of five constituents in Salvia miltiorrhiza phenolic acids extract by wavelength switching

目的建立高效液相色谱法测定丹参酚酸提取物中丹参素钠、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B。方法采用Phenomsil-C18(250 mm×4.6 mm,5μm)色谱柱,流动相为乙腈-0.5%磷酸水溶液,梯度洗脱,体积流量1.0mL/min,检测波长为280 nm(0～50 min)、330 nm(50～68 min)、286nm(68～90 min),柱温30℃。结果丹参素钠、原儿茶醛、迷迭香酸、紫草酸、丹酚酸B的进样量分别在0.033 96～0.339 6μg、0.016 60～0.166 0μg、0.092 4～0.924μg、0.084 7～0.847μg、1.300～13.00μg范围内与色谱峰面积呈良好的线性关系;加样回收率(n=6)均在99.2%～100.2%,RSD均小于2.3%。7个来源的丹参酚酸提取物中5种物质的含有量有一定差异。结论本法快速、准确,重复性好,可更好地控制丹参酚酸提取物的质量。

AIM To establish an HPLC method for simultaneously determining five constituents（sodium danshensu,protocatechuic aldehyde,rosmarinic acid,lithospermic acid,salvianolic acid B） in Salvia miltiorrhiza phenolic acids extract.METHODS The separation was performed on a Phenomsil-C18 ODS（250 mm×4.6 mm,5 μm） column with the gradient elution of acetonitrile-water of 0.5% phosphoric acid at the flow rate of 1.0 mL/min.The column temperature was set at 30 ℃.Detection wavelength was 280 nm in 0-50 min,330 nm in 50-68 min,286 nm in 68-90 min.RESULTS Sodium danshensu,protocatechuic aldehyde,rosmarinic acid,lithospermic acid and salvianolic acid B had good linearity in the ranges of 0.033 96-0.339 6 μg,0.016 60-0.166 0 μg,0.092 4-0.924 μg,0.084 7-0.847 μg,1.300-13.00 μg,repectively.The average recoveries of constituents were 99.2%-100.2%,with RSDs less than 2.3%.The contents of five constituents in Salvia miltiorrhiza phenolic acids extract from seven sources showed differences.CONCLUSION The developed method is rapid,accurate with high roproducibility and stability,and helpful to control the quality of Salvia miltiorrhiza phenolic acids extract.