高效液相色谱变波长法同时测定杜仲平压片中绿原酸和芦丁的含量

Simultaneous content determination of chlorogenic acid and rutin in Duzhong Pingya Pian(杜仲平压片)by HPLC with variable wavelength

目的建立杜仲平压片中绿原酸和芦丁的含量测定方法。方法采用高效液相色谱(HPLC)变波长法同时测定杜仲平压片中绿原酸和芦丁的含量,色谱柱为Waters sunfire-C18(4.6 mm×250.0 mm,5μm),流动相为乙腈(A)-0.5%磷酸水溶液(B),梯度洗脱(0~11 min,12%A;>11~12 min,12%→24%A;>12~20 min,24%A);检测波长327 nm(0~12 min)、354 nm(>12~20 min);流速1.0 mL/min,柱温35℃,进样量10μL。结果绿原酸回归方程为Y=27.739X-2.015(r=0.9998),芦丁回归方程为Y=15.508X-0.0584(r=0.9997),表明绿原酸和芦丁分别在18.66~261.24,1.04~14.57μg/mL范围内呈良好的线性关系;绿原酸和芦丁的平均回收率分别为103.76%,98.38%,RSD分别为0.70%,0.94%。结论该法简便可行、重现性好,可用于杜仲平压片中绿原酸和芦丁的含量测定。

Objective To establish a method of content determination of chlorogenic acid and rutin in Duzhong Pingya Pian(杜仲平压片)by HPLC with variable wavelength.Methods HPLC with variable wavelength was carried out on Waters sunfire-C18 column(4.6 mm×250.0 mm,5μm),with a mixture of acetonitrile(A)and 0.5%phosphoric acid(B)as the mobile phase;with gradient elution(0-11 min,12%A;>11-12 min,12%→24%A;>12-20 min,24%A),and the detection wavelengths were 327 nm(0-12 min),354 nm(>12-20 min);at a flow rate of 1.0 mL/min,the column temperature was set at 35℃,the sample size was 10μL.Results The regression equation of chlorogenic acid was Y=27.739X-2.015(r=0.9998),that of rutin was Y=15.508X-0.0584(r=0.9997),which showed that a good linearity of chlorogenic acid and rutin was in the range of 18.66-261.24 and 1.04-14.57μg/mL,respectively.The average recovery rate of chlorogenic acid and rutin was 103.76%and 98.38%,respectively.And RSD was 0.70%and 0.94%,respectively.Conclusion The method is simple,feasible and repeatable,and can be applied in the content determination of chlorogenic acid and rutin in Duzhong Pingya Pian.