大孔吸附树脂纯化黄芩中三种黄酮的含量测定

Determination of Three Kinds of Flavonoids in Scutellaria Baicalensis Purified by Macroporous Absorption Resin

关键词:  高效液相色谱  大孔吸附树脂  黄芩苷  黄芩素  汉黄芩素

Key words:  HPLC  macroporous absorption resin  baicalin  baicalein  wogonin

摘要:

目的 建立高效液相色谱法同时测定大孔吸附树脂富集纯化后的黄芩提取物中黄芩苷、黄芩素和汉黄芩素含量的方法。方法 以AB-8型大孔吸附树脂对黄芩水提取物中总黄酮进行富集纯化，采用高效液相色谱法同时测定其中黄芩苷、黄芩素和汉黄芩素的含量。色谱柱：Agilent TC-C₁₈(4.6 mm×250 mm，5 μm)；流动相：乙腈-0.1%磷酸水溶液，梯度洗脱；检测波长：280 nm；流速1.0 mL·min^-1；柱温：30 ℃。结果 黄芩苷、黄芩素和汉黄芩素分别在0.022 4~0.448 0 μg (r=0.999 9)，0.021 0~0.420 0 μg(r=0.999 9)，0.010 3~0.205 0 μg(r=0.999 9)内线性关系良好；平均回收率分别为97.3%(RSD 1.71%)，98.4%(RSD 2.05%)，98.1%(RSD 1.65%)。黄芩水提取物经大孔吸附树脂纯化后，黄芩苷、黄芩素和汉黄芩素的平均含量分别为27.52%、6.87%和2.13%，未经纯化的黄芩水提取物中三者的平均含量分别为9.16%、1.66%和0.54%。结论 本方法简便、准确、灵敏度高，重复性好。

Abstract:

OBJECTIVE To establish a method for determination of the content of baicalin, baicalein and wogonin in Scutellaria baicalensis purified by macroporous absorption resin by HPLC. METHODS The macroporous absorption resin AB-8 was used to separate and purify the total flavonoids in the water extract from Scutellaria baicalensis. The HPLC method was used to determine the content of baicalin, baicalein and wogonin. The analytical column was Agilent TC-C₁₈(4.6 mm×250 mm, 5 μm). The mobile phase was a mixture liquid of acetonitrile and 0.1% phosphoric acid in water by gradient elution. The UV detector wavelength was 280 nm, the flow rate was 1.0 mL·min^-1, and the column temperature was 30 ℃. RESULTS The linear ranges of baicalin, baicalein and wogonin were 0.022 4-0.448 0 μg(r=0.999 9), 0.021 0-0.420 0 μg(r=0.999 9), 0.010 3-0.205 0 μg(r=0.999 9), respectively. The average recoveries of the method for baicalin, baicalein and wogonin were 97.3% with RSD 1.71%, 98.4% with RSD 2.05% and 98.1% with RSD 1.65%, respectively. The average contents of baicalin, baicalein and wogonin in the water extract from Scutellaria baicalensis with macroporous absorption resin were 27.52%, 6.87% and 2.13%, and the average contents were 9.16%, 1.66% and 0.54% without purification. CONCLUSION The method is simple, accurate, sensitive and stable.