HPLC法同时测定葛根中5种异黄酮的含量

Simultaneous determination of 5 isoflavones in Radix puerariae by HPLC

目的建立能同时测定葛根中5种异黄酮含量的HPLC法。方法采用Phenomenex C 18色谱柱(250 mm×4.6 mm,5μm);柱温:40℃;流动相A为甲醇,B为水,进行梯度洗脱;流速为1.0 mL·min^-1;检测波长为260 nm。结果葛根素、大豆苷、染料木苷、大豆苷元和染料木素的回归方程分别为y 1=35462.65 x 1+899.4(r 1=0.9999),y2=89343.6 x2-720.2(r2=0.9999),y 3=139651.625 x3+1351.7(r 3=0.9998),y4=129138.175 x4-1339.5(r 4=0.9999)和y5=216079.6 x5-3679.8(r5=0.9999),分别在0.015~0.150,0.048~0.480,0.049~0.490,0.049~0.490和0.052~0.520μg范围内线性关系良好,平均回收率分别为98.3%,99.6%,97.5%,100.1%和96.0%。结论该方法同时测定了葛根中5种异黄酮的含量,结果准确、可靠,为葛根质量评价奠定了基础。

Objective To establish an HPLC method for the determination of 5 isoflavones in Radix puerariae.Methods The Phenomenex C 18 chromatographic column(250 mm×4.6 mm,5μm)was used;column temperature was 40℃;the mobile phase A was methanol and B was water,and gradient elution was performed.The flow rate was 1.0 mL·min^-1 and the detection wavelength was 260 nm.Results The regression equation of puerarin in the range of 0.015-0.150μg was y 1=35462.65 x 1+899.4(r 1=0.9999),and the average recovery was 98.3%.The regression equation of daidzin in the range of 0.048-0.480μg was y 2=89343.6 x 2-720.2(r 2=0.9999),and average recovery was 99.6%.The regression equation of genistin in the range of 0.049-0.490μg was y 3=139651.625 x 3+1351.7(r 3=0.9998)and the average recovery was 97.5%.The regression equation of dsupportzein in the range of 0.049-0.490μg was y 4=129138.175 x 4-1339.5(r 4=0.9999),and the average recovery was 100.1%.The regression equation of genistein in the range of 0.052-0.520μg was y 5=216079.6 x 5-3679.8(r 5=0.9999),and the average recovery was 96.0%.Conclusion The method was accurate and reliable for the determination of 5 isoflavones in Radix puerariae.The results were accurate and reliable,which provided a basis for the quality evaluation of Radix puerariae.