UV和HPLC法测定黄芩黄酮总苷元提取物中总黄酮和3种主要成分的含量

Determination of Total Flavonoids and Three Main Components in Scutellariae Radix Total Flavone Aglycone Extract by Ultraviolet Spectrophotometry and High-performance Liquid Chromatogaphy

本文建立了黄芩黄酮总苷元提取物中总黄酮和3种主要成分的含量测定方法。采用HPLC法测定黄芩素、汉黄芩素和千层纸素A的含量。使用Agilent 1200型高效液相色谱仪，Eclipse XDB-C18（250 mm×4.6 mm，5 μm）色谱柱，流动相为甲醇-0.2%磷酸水（50：50），流速1 mL / min，柱温35 ℃，检测波长275 nm的方法。黄芩素、汉黄芩素和千层纸素 A线性范围分别为3.501-70.016 μg/mL（r=0.9999），0.832-16.640 μg/mL（r=0.9998），0.418-8.352 μg/mL（r=0.9999）；平均加样回收率（n=6）分别为99.55%、98.86%、9.997%，RSD分别为1.74%、1.74%、1.70%。采用UV法测定总黄酮含量，黄芩素线性范围为1.71-8.54 μg/mL（r=0.9991）；平均加样回收率（n=6）为100.93%，RSD为2.63%。黄芩素、汉黄芩素、千层纸素 A和总黄酮的含量分别为48.22%-60.36%、12.09%-14.46%、5.05%-8.81%、78.11%-90.12%；研究结果表明该方法准确、可靠，适用于黄芩黄酮总苷元提取物的含量测定。

The paper established a method for separation and determination of total flavonoids and three main components in Scutellaria Radix total flavone aglycone extract by ultraviolet （UV） spectrophotometry and high-performance liquid chromatography.The anlysis was performed on an Agilent 1200 Series HPLC system with Eclipse XDB-C18 （250 mm×4.6 mm,5 μm） column.The mobile phase consisted of methanol-0.2% phosphoric acid water （50：50） with the flow rate of 1 min/mL.The column remained at 35 ℃ and the detection wavelength was set at 275 nm.The linear ranges of baicalein,wogonin and oroxylin A were 3.501-70.016 μg/mL （r=0.9999）, 0.832-16.640 μg/mL （r=0.9998） and 0.418-8.352 μg/mL （r=0.9999）,respectively.The average recoveries （n=6） were 99.55% （RSD=1.74%）,98.86% （RSD=1.74%） and 99.97% （RSD=1.70%）.Total flavonoids were determined by UV spectrometry.The linear range of baicalein was 1.71-8.54 μg/mL （r=0.9991）.The average recoveries （n=6） were 100.93% （RSD=2.63%）.The contents of baicalein,wogonin,oroxylin A and total flavonoids were 48.22%-60.36%,12.09%-14.46%, 5.05%-8.81% and 78.11%-90.12%,respectively.