HPLC法同时测定青天葵药材中7种黄酮的含量

Simultaneous determination of seven flavonoids in Nervilia fordii with HPLC

建立同时测定青天葵中鼠李秦素(1)、鼠李柠檬素(2)、鼠李素(3)、鼠李秦素-3-O-β-D-葡萄糖苷(4)、鼠李秦素-3-O-β-D-木糖-(1→4)-β-D-葡萄糖苷(5)、鼠李秦素-3-O-β-D-葡萄糖-(1→4)-β-D-葡萄糖苷(6)和鼠李柠檬素-3-O-β-D-葡萄糖-(1→4)-β-D-葡萄糖苷(7)含量的高效液相色谱法。采用Kromasil C18色谱柱(250 mm×4.6mm,5μm),以0.4%磷酸乙腈为流动相进行梯度洗脱,流速为1.0 mL.min 1,检测波长为256 nm,柱温为40℃。7种黄酮类化合物(1～7)的线性范围分别为0.55～70.00μg.mL 1(r=0.999 7)、0.86～110.00μg.mL 1(r=0.999 7)、0.39～50.00μg.mL 1(r=0.999 7)、0.55～70.00μg.mL 1(r=0.999 5)、1.33～170.00μg.mL 1(r=0.999 8)、1.33～170.00μg.mL 1(r=0.999 8)、0.16～20.00μg.mL 1(r=0.999 5),平均回收率在97.19%～99.45%之间,RSD在0.91%～2.69%之间。该方法简单、准确,具有良好的重复性和稳定性,可为青天葵的质量控制提供科学依据。

The study is to develop an HPLC method for simultaneous determination of rhamnazin（1）,rhamnocitrin（2）,rhamnetin（3）,rhamnazin-3-O-β-D-glucopyranoside（4）,rhamna-zin-3-O-β-D-xylopyranosyl-（1→4）-β-D-glucopyranoside（5）,rhamna-zin-3-O-β-D-glucopyranosyl-（1→4）-β-D-glucopyranoside（6）,and rhamnocitrin-3-O-β-D-glucopyranosyl-（1→4）-β-D-glucopyranoside（7） in Nervilia fordii.The separation was performed on a Kromasil C18 column（250 mm × 4.6 mm,5 μm） with 0.4% phosphoric acid acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL min 1.The detect wavelength was set at 256 nm,and the column temperature was set at 40 ℃.There were good linear relationships between the logarithm values of concentrations and those of the peak areas of seven flavonoids（1 7） in the range of 0.55 70.00 μg mL 1（r = 0.999 7）,0.86 110.00 μg mL 1（r = 0.999 7）,0.39 50.00 μg mL 1（r = 0.999 7）,0.55 70.00 μg mL 1（r = 0.999 5）,1.33 170.00 μg mL 1（r = 0.999 8）,1.33 170.00 μg mL 1（r = 0.999 8）,0.16 20.00 μg mL 1（r = 0.999 5）,respectively.The recoveries of the seven flavonoids were between 97.19 %-99.45%,the relative standard deviations（RSDs） were between 0.91 %-2.69%.The established method is rapid,accurate with high re-peatability,which could provide scientific evidence for the quality control of Nervilia fordii.