HPLC、UPLC测定芪参益气滴丸中5个丹参活性成分含量的比较研究

Comparison of HPLC and UPLC determination of five active components of Radix et Rhizoma Salviae Miltiorrhizae in Qishenyiqi dropping pills

目的:建立同时测定芪参益气滴丸中5个丹参活性成分(丹参素、原儿茶醛、迷迭香酸、丹酚酸B、丹酚酸A)含量的高效液相色谱(HPLC)和超高效液相色谱(UPLC)方法,并对建立的方法进行比较。方法:HPLC方法色谱柱为Agilent Zorbax SBC18柱(4.6 mm×250 mm,5μm),UPLC方法色谱柱为Waters Acquity BEH C18柱(2.1 mm×100 mm,1.7μm),流动相均为乙腈-甲酸-水体系梯度洗脱,2种方法的流速分别为1.0和0.4 mL.min-1,检测波长均为280 nm,柱温均为30℃。比较2种方法的分离情况及线性、精密度、重复性、加样回收率、样品测定结果。结果:UPLC的分析时间少于HPLC;5个物质在一定范围内均呈现良好线性,r均大于0.9998;2种方法的精密度、重复性、加样回收率和样品测定结果无显著性差异。结论:HPLC和UPLC 2种方法准确性、重复性均较好;UPLC分析比HPLC具有更加快速、高效的优势。

Objective：To compare the established high performance liquid chromatography（HPLC） and ultra-performance liquid chromatography（UPLC） method for simultaneous determination of five active compounds in Qishenyiqi dropping pills（danshensu,protocatechualdehyde,rosmarinic acid,salvianolic acid B and salvianolic acid A）.Methods：A gradient HPLC system was used in the quantitative analysis of five compounds on the reversed-phase column of Agilent Zorbax SB C18（4.6 mm×250 mm,5 μm）,using acetonitrile-water-formic acid as mobile phase,with a flow rate of 1.0 mL·min-1.A Waters Acquity BEH C18 column（2.1 mm×100 mm,1.7 μm） was applied,using the same mobile phase as the HPLC method,with a flow rate of 0.4 mL·min-1 in UPLC.The detective wavelength was set at 280 nm and the column temperature was 30 ℃ in both of the methods.The five active compounds were determined by HPLC and UPLC,respectively.In order to compare the two methods,the separation resolution,linearity,repeatability,recovery and the contents of the five compounds were investigated.Results：The analysis time of UPLC was obviously shorter than that of HPLC.The linear relationship between the concentration and peak areas of the five compounds were all linear（r0.9998）.There were no significant difference among precision,recovery,repeatability and the contents of the two methods.Conclusion：Both of the methods show good accuracy and repeatability.Compared with HPLC,UPLC is more fast and high-performance.