高效和超高效液相色谱法测定乐脉丸中丹酚酸B含量的比较

摘要

目的 运用高效液相色谱法和超高效液相色谱法对乐脉丸中丹酚酸B的含量进行测定,并对测定方法和结果进行比较.方法 高效液相色谱法中,色谱柱采用迪马C18柱(250 mm×4.6mm,5 μm),流动相为甲醇-乙腈-17％甲酸(27:4:69),流速为1.0 mL/min,检测波长为286 nm,进样量为5μL.超高效液相色谱法中,色谱柱采用Acquity UPLC BEH C18柱(50 mm×2.1mm,1.7 μm),流动相为甲醇-乙腈-17％甲酸(20:1:79),流速为0.6 mL/min,进样量为1μL.结果 高效液相色谱法主峰出峰时间为45 min,3批样品含量分别为16.94,12.06,6.84 mg/g;超高效液相色谱法主峰出峰时间为1 min,3批样品含量分别为16.93,12.06,6.84 mg/g.结论 两种方法均可用于测定乐脉丸中丹酚酸B的含量,与高效液相色谱法相比,超高效液相色谱法加快了分析速度,提高了分析效率,减少了有机溶剂的使用.%Objective To determine the content of salvianolic acid B in Lemai Pill by the HPLC and UPLC methods and to compare the detecting methods and their detected results. Methods In the HPLC method, the Diamonsil C18 column(250 mm×4. 6 mm, 5 μm) was used with the mobile phase of methanol - acetonitrile -17% formic acid (27 : 4 : 69) at a flow rate of 1 mL/min,the injection volume was 5 μL,and the detective wavelength was set at 286 nm. In the UPLC method,AcquityUPLC BEH C18 column(50 mm×2. 1 mm,1.7 μm) was used with the mobile phase of methanol - acetonitrile - 17% formic acid (20 : 1 :79) at a flow rate of 0.6 mL/min,and the injection volume was 1 μL. Results The appearance time of the main peak in HPLC was 45 min,and the contents of salvianolic acid B in three batches of samples were 16.94,12.06,6.84 mg/g,respectively. The appearance time of the main peak in UPLC was 1 min.and the contents of salvianolic acid B in three batches of samples were 16.93,12.06,6.84 mg/g,respectively. Conclusion Both two methods can be adopted to accurately determine the content of salvianolic acid B in Lemai Pill. Compared with the HPLC method,the UPLC method accelerates the analytic speed, increases the analytic efficiency and reduces the use amount of organic solvents.