HPLC法同时测定葛根芩连汤中8种有效成分的含量

目的: 建立运用HPLC法同时测定葛根芩连汤中8种有效成分(葛根素、甘草苷、黄芩苷、汉黄芩苷、巴马汀、小檗碱、黄芩素、汉黄芩素)的含量。方法: 采用反相高效液相色谱法测定,色谱柱:Agilent ZORBAX Eclipse SB-C₁₈(4.6 mm×250 mm,5 μm);流动相:甲酸铵(pH 3.0、10 mmol·L^-1)-乙腈,梯度洗脱;流速:1 mL·min^-1;检测波长:270、360 nm;柱温:25 ℃;进样量:20 μL。结果:葛根素、甘草苷、黄芩苷、汉黄芩苷、巴马汀、小檗碱、黄芩素、汉黄芩素的质量浓度分别在1.12~22.3 μg·mL^-1(r=0.9990),1.08~21.7 μg·mL^-1(r=0.9991),4.10~82.0 μg·mL^-1(r 0.9990),1.23~24.6 μg·mL^-1(r=0.9993),1.99~39.8 μg·mL^-1(r=0.9991),4.05~81.0 μg·mL^-1(r=0.9993),2.40~48.0 μg·mL^-1(r=0.9994),1.11~22.1 μg·mL^-1(r =0.9993)范围内与色谱峰面积呈良好的线性关系。平均回收率(n=5)均在95%~105%范围内,RSD均小于3.0%。结论: 本方法可快速简便测定葛根芩连汤中8个有效成分的含量。

Objective: To establish an HPLC method to determine eight active components in Gegen Qinlian Decoction simultaneously. Methods: HPLC, column:Agilent ZORBAX Eclipse SB-C18 column(4.6 mm × 250 mm, 5 μm), mobile phase:HCOONH₃(pH3.0, 10 mmol·L^-1)-CH₃CN, flow rate:1.0 mL·min^-1, detection wavelength:270 nm and 360 nm, temperature:25 ℃, injection volume:20 μL. Results: The method has a good linearity in the ranges of 1.12-22.3 μg·mL^-1(r =0.9990)for puerarin, 1.08-21.7 μg·mL^-1(r =0.9991)for liquiritin, 4.10-82.0 μg·mL^-1(r =0.9990)for baicalin, 1.23-24.6 μg·mL^-1(r =0.9993)for wogonoside, 1.99-39.8 μg·mL^-1(r = 0.9991)for palmatine, 4.05-81.0 μg·mL^-1(r =0.9993)for berberine, 2.40-48.0 μg·mL^-1(r =0.9994)for baicalein, 1.11-22.1 μg·mL^-1(r =0.9993)for wogonin.The average recoveries(n=5)of eight active compounds were 95%-105 % with RSDs of recoveries(n=5)were less then 3.0% Conclusion: The method for determination of 8 active compounds in Gegen Qinlian Decoction, this method can be used for rapid and convenient evaluation of the quality of Gegen Qinlian Decoction.