高效液相色谱法同时测定丹参中10种水溶性和4种脂溶性成分的含量

目的: 建立高效液相色谱法同时测定不同来源的丹参药材及饮片中丹参素钠、原儿茶醛、咖啡酸、异阿魏酸、丹酚酸D、迷迭香酸、紫草酸、丹酚酸B、丹酚酸A、丹酚酸C、二氢丹参酮Ⅰ、隐丹参酮、丹参酮Ⅰ、丹参酮Ⅱ_A 10种水溶性化合物和4种脂溶性化合物的含量。方法: 采用Agilent Zorbax SB-aq(250 mm×4.6 mm,5 μm)色谱柱,以乙腈-0.03%磷酸溶液为流动相,梯度洗脱(0~60 min,10%A→68%A;60~70 min,68%A→80%A),流速0.8 mL·min^-1,柱温30 ℃,检测波长280 nm,进样量20 μL。结果: 在65 min内,丹参中的14种成分可实现完全分离,质量浓度在0.01~1 766.67 μg·mL^-1范围内与峰面积呈良好的线性关系,相关系数r不低于0.999 5;平均回收率(n=6)为97.4%~102.5%,RSD<2.6%。不同的产地来源、叶子大小、根茎粗细、生长年限及栽培方式等因素对丹参中指标性成分含量差异都有很大的影响。结论: 该分析方法专属性好,灵敏度高,定量准确,经方法学验证可用于同时测定丹参中的10种水溶性和4种脂溶性化合物的含量,为丹参药材及饮片的质量评价标准提供参考。

Objective: To establish an HPLC method for the simultaneous determination of ten hydrophilic compounds and four lipophilic compounds,i.e.,sodium danshensu,protocatechualdehyde,caffeic acid,isoferulic acid,salvianolic acid D,rosmarinic acid,lithospermic acid,salvianolic acid B,salvianolic acid A,salvianolic acid C,1,2-dihydrotanshinquinone,cryptotanshinone,tanshinone Ⅰ,tanshinone Ⅱ_A in Radix Salviae Miltiorrhizae from different sources in order to provide the foundation for the improvement of quality standard of Radix Salviae Miltiorrhizae. Methods: The HPLC separation was performed on an Agilent Zorbax SB-aq(250 mm×4.6 mm,5 μm)column eluted with the mobile phase of acetonitrile-0.03% phosphoric acid solution.The separation was achieved using the following gradient:0-60 min,10%A→68%A;60-70 min,68%A→80%A,at a flow rate of 0.8 mL·min^-1.The column temperature was controlled at 30 ℃,the UV detection wavelength was 280 nm,and the injection volume was 20 μL. Results: The complete separation was obtained within 65 minutes for the 14 compounds.The linear ranges of the 14 compounds were between 0.01-1 766.67 μg·mL^-1 and the correlation coefficients were more than 0.999 5.The average recoveries of the 14 compounds were 97.4%-102.5% and RSDs were less than 2.6%.There was a great difference in the contents of 14 compounds in Radix Salviae Miltiorrhizae as a result of different sources,different leaf sizes,different rhizome thickness,different growth ages and different cultivation methods. Conclusion: The proposed method is selective,sensitive,accurate and validated,which can be used for simultaneous determination of ten hydrophilic compounds and four lipophilic compounds in Radix Salviae Miltiorrhizae.This method can provide reference for evaluating the quality of Radix Salviae Miltiorrhizae.