高效液相法测定颐神达络方中葛根素、3′-甲氧基葛根素、芹糖基葛根素、大豆苷和芍药苷的含量

Quantitative Determination of puerarin, 3′-methoxyl-puerarin, mirificin, daidzin and paeoniflorin in Yishendaluofang by RP-HPLC

目的建立反相高效液相法测定颐神达络方中葛根素、3′-甲氧基葛根素、芹糖基葛根素、大豆苷和芍药苷含量的方法.方法采用ODS HYPERSIL 色谱柱（250 mm×4.6 mm,5 μm）,乙腈-4‰磷酸水梯度洗脱,检测波长为240 nm,柱温：室温,对葛根素、3′-甲氧基葛根素、芹糖基葛根素、大豆苷和芍药苷的含量进行测定.结果葛根素、3′-甲氧基葛根素、芹糖基葛根素、大豆苷和芍药苷分别在0.152～0.912 μg、0.184～1.104 μg、0.240～1.200 μg、0.142～0.852 μg、0.204～1.224 μg的范围内呈现良好的线性关系,r=0.999 5、0.999 6、0.999 3、0.999 6、0.999 7（n=6）,平均加样回收率分别为100.09%、101.12%、100.69%、100.05%、99.19%,RSD分别为2.00%、2.97%、2.90%、2.95%、1.62%.结论本方法简便、准确、可靠,可以作为颐神达络方有效部位的质量控制方法.

Objective To establish a method for quantitative determination of puerarin, 3＇-methoxyl- puerarin, mirificin, daidzin and paeoniflorin in Yishendaluo Formula by RP-HPLC. Method The RP- HPLC procedure was used in the determination as follows： the chromatographic column was ODS HYPERSIL （250 mm ×4.6 mm,5μm）, the gradient elution program was acetonitrile： 4%0 H3PO4 water, the detection wavelength was 240 nm, and the column temperature was the room temperature. Results The results showed that puerarin, 3＇-methoxyl-puerarin, mirificin, daidzin and paeoniflorin had a good linear relationship respectively when the concentration was between 0. 152 μg - 0. 912 μg, 0. 184 μg - 1. 104 μg, 0.240 μg -1.200 μg, 0. 142 μg -0.852μg, and 0.204μg -1.224 μg respectively （r =0. 999 5s, 0.999 6, 0.999 3, 0.999 6 and 0.999 7 respectively, n =6）. The average recoveries of the samples were 100.09%, 101.12%, 100.69%, 100.05% and 99.19% respectively, RSDs were 2. 00%, 2.97%, 2.90%, 1.62% and 2.95% respectively. Conclusion The procedure, which is easy, accurate and reliable, can be used to control the quality of the effective fraction of Yishendaluo Formula.