HPLC测定2种大黄中4类功效组分含量

Determination of Contents of Four Types of Active Constituents in Rhei Radix ex Rhizoma by HPLC

摘要:

目的: 建立用高效液相色谱法快速测定2种大黄游离型蒽醌、结合型蒽醌、番泻苷、鞣质单体4类14种与功效紧密相关组分含量方法,为全面评价大黄药材质量提供依据。 方法: 采用高效液相色谱法测定,Agilent Zorbax SB-C₁₈色谱柱(4.6 mm×250 mm,5 μm);流动相Ⅰ:0.1%磷酸水-甲醇梯度洗脱,流动相Ⅱ:四氢呋喃-水-冰醋酸(2∶80∶1.5)与乙腈梯度洗脱,流动相Ⅲ:0.5%冰醋酸水-甲醇梯度洗脱;流速分别为1.0,0.8,1.0 mL·min^-1,检测波长分别为254,350,270 nm,柱温25 ℃。 结果: 测定了两种大黄的4类功效组分含量,各组分均在30 min内均得到良好分离;不同成分在各自的线性范围内线性关系良好,平均回收率均在97.09%~103.2%,RSD均在0.15%~3.0% (n=6)。 结论: 方法简便、准确、重复性好,可作为大黄全面质量控制。

Abstract:

Objective:To establish quantification methods for determination of four types of active constituents in two sorts of Rhei Radix ex Rhizoma by HPLC, including free anthraquinone, anthraquinone glycoside, sennoside, catechin and gallic acid, in order to evaluate the quality of Rhei Radix ex Rhizoma all-around. Method: HPLC was applied on the determination of four types of active constituents with Agilent Zorbax SB-C₁₈ column (4.6 mm×250 mm,5 μm). Mobile phaseⅠwas 0.1% phosphoric acid-methanol by gradient elution, mobile phaseⅡwas THF-water-acetic acid(2∶80∶1.5)-acetonitrile by gradient elution, and mobile phase Ⅲ was 0.5% acetic acid-methanol by gradient elution; the three kinds of flow rate were 1.0,0.8,1.0 mL·min^-1; the wavelengths were set at 254,350,270 nm; the column temperature were all set at 25 ℃. Result: Determine of four types of active constituents in two different sorts of Rhei Radix ex Rhizoma. These components were separated completely within 30 minutes under this chromatographic condition. The average recoveries were from 97.09% to 103.2%, its relative standard deviations were from 0.15% to 3.0% (n=6). Conclusion: The methods with good reproducibility are simple and accurate; it can be used as quality control methods for Rhei Radix ex Rhizoma.