高效液相色谱法同时测定葛根提取物中5种异黄酮的含量

HPLC simultaneous determination of five isoflavonoids in Radix Pueraria extracts

目的:建立葛根提取物中5种异黄酮含量测定的HPLC方法。方法:采用Hypersil C18色谱柱(250 mm×4.6 mm,5μm),以乙腈-0.02 mol·L-1醋酸铵(醋酸调pH=4.6)为流动相,进行梯度洗脱,流速为1.0mL·min-1,检测波长为260 nm,柱温为35℃。结果:葛根素、大豆苷、染料木苷、大豆素、染料木素分别在0.204～510.4μg·mL-1(r=0.999 9),0.201～503.0μg·mL-1(r=0.999 9),0.233～581.8μg·mL-1(r=0.999 9),0.201～502.4μg·mL-1(r=0.999 9),0.195～487.6μg·mL-1(r=0.999 9)范围内线性关系良好。平均回收率(n=6)分别为101.52%(RSD=0.69%),102.66%(RSD=0.30%),98.90%(RSD=0.26%),98.64%(RSD=0.29%),100.16%(RSD=0.20%)。结论:本方法快速、简便、精密度和重现性好、灵敏度高,可作为葛根提取物的含量控制方法。

OBJECTIVE To develop a HPLC method for the determination of the five isoflavonoids in Radix Pueraria extracts. METHODS The chromatography was performed on a Hypersil C18 column （250 mm× 4. 6 mm, 5 μtm）with a mobile phase of acetonitrile - 0. 02 mol·L^-1 ammonium acetate （adjusted to pH 4. 6 with acetic acid）and gradient elution. The flow rate was 1.0 mL.min^-1. The column temperature was 35 ℃. The detection wavelength was 260 nm. RESULTS The calibra tion curves of puerarin, daidzin, genistin, daidzein, genistein were linear in the range of 0.204-510.4μg·mL^-1（r=0.9999）,0.201-503.0μg·mL^-1（r=0.9999）,0.233-581.8μg·mL^-1（r=0.9999）,0.201-502.4μg·mL^1-（r=0.9999）,0.195-487.6μg·ml^-1（r=0.9999）, respectively. The mean recoveries （n = 6） were 101.52%（RSD=0.69%）,102.66%（RSD7=0.30%）,98.90%（RSD=0.26%）,98.64%（RSD=0.29%）,100.16%（RSD=0.20%）, respectively. CONCLUSION The method is simple, rapid, accurate, reproducible and sensitive. It can be used as the content control method for Radix Pueraria extracts.