反相高效液相色谱法同时测定丹参中5种成分含量

A Simultaneous Determination of Five Components in Radix et Rhizoma Salviae Miltiorrhizae by RP-HPLC

目的建立高效液相色谱法同时测定丹参中3种水溶性和2种脂溶性成分。方法采用反相高效液相色谱法,WelchromC18(250 mm×4.6 mm,5μm)色谱柱,以乙腈(A)-0.1%磷酸(B)为流动相梯度洗脱:0~15 min,10%→12%A;~35 min,12%→20%A;~45 min,20%→60%A;~65 min,60%→65%A;~80 min,65%→80%A;~90 min,10%A。流速1 m L·min-1,检测波长280 nm,柱温30℃。结果丹参素、迷迭香酸、丹酚酸B、隐丹参酮及丹参酮ⅡA分别在0.0595~2.380 0,0.346 0~13.840 0,0.656 0~26.240 0,0.420 0~16.800 0,0.414 0~16.560 0μg范围内线性良好,r=0.999 9,加样平均回收率98.69%~100.91%,RSD均<1.2%(n=6)。结论该方法快速、准确、重复性好,可用于同时测定丹参中水溶性和脂溶性成分,较全面控制丹参质量。

Objective To develop a method for simultaneous determination of three hydrophilic components and two lipophilic components in Radix et Rhizoma Salviae Miltiorrhizae.Methods The RP-HPLC method was performed by using a Welchrom C18column(250 mm×4.6 mm,5 μm) with a mobile phase of acetonitrile(A)-0.1%phosphoric acid(B).The gradient elution program was as follows:0-15 min,10%→12%A;-35 min,12%→20%A;-45 min,20%→60% A;-65 min,60% →65%A;-80 min,65% →80% A;-90 min,10% A.The flow rate was kept at 1.0 m L · min-1.The detection wavelength was set at280 nm.The column temperature was 30 ℃.Results A good linearity was obtained over 0.059 5-2.380 0 μg for tanshinol,0.346 0-13.840 0 μg for rosmarinic acid,0.656 0-26.240 0 μg for salviamolic acid B,0.420 0-16.800 0 μg for cryptotanshinone and 0.414 0-16.560 0 μg for tanshinone IIA,respectively(r = 0.999 9).The average recovery rates were between 98.69%-100.91% with RSD less than 1.2%(n = 6).Conclusion The method is rapid,accurate,credible and repeatable,and can provide basis for the quality control of Radix et Rhizoma Salviae Miltiorrhiza.