高效液相色谱法测定蒿板青颗粒中一枝蒿酮酸及 (R, S)-告依春的含量

Determination of Rupestonic Acid and (R, S)-epigoitrin in Haobanqing granules by HPLC

关键词:  高效液相色谱法  蒿板青颗粒  一枝蒿酮酸  (R, S)-告依春  含量测定

Key words:  HPLC  Haobanqing granules  Rupestonic acid  (R, S)-epigoitrin  Content determination

摘要:

为有效控制蒿板青颗粒的质量，采用高效液相色谱法对蒿板青颗粒中有效成分一枝蒿酮酸和 (R, S) -告依春含量进行了研究。采用C18柱（250 mm×4.6 mm，5 μm)，流动相为乙腈（A）- 0.4%磷酸水溶液（B），梯度洗脱（0 min，5% A；0～10min，5% A→15% A；10～25min，15% A→30% A；25～40min，30% A；40～41min，30% A→5% A；41～45min，5% A），检测波长为245 nm，流速1.0mL/min，柱温35℃。结果表明，在该色谱条件下，辅料对一枝蒿酮酸和 (R,S) -告依春出峰均无干扰，一枝蒿酮酸和 (R, S) -告依春与相邻杂质峰分离良好；一枝蒿酮酸、(R, S)-告依春浓度分别在24～120 μg /mL和4.8～24 μg /mL范围内，峰面积与其对应的含量呈现良好的线性关系（一枝蒿酮酸R2 = 0.9992，3组样品中一枝蒿酮酸的平均回收率分别为98.4%、97.9%和98.1%，RSD分别为1.31%、1.02%和1.33%；(R, S)-告依春R2 = 0.9997，3组样品中(R, S)-告依春的平均回收率分别为98.8%、98.3%和97.5%，RSD分别为0.56%、0.88%和1.14%）。该方法简便、分析快速、准确度高，重复性好，适用于蒿板青颗粒中有效成分一枝蒿酮酸和(R, S)-告依春的质量控制。

Abstract:

In order to improve the quality standard of Haobanqing granules, the contents of rupestonic acid and (R, S)-epigoitrin in Haobanqing granules was determined by HPLC. The isolation was performed on a C18 column ( 4.6 mm × 250 mm, 5 μm) with mobile phase consisting of acetonitrile (A)–0.4% phosphoric acid aqueous solutions(B) using gradient elution (0min, 5% A; 0~10min, 5% A→15% A; 10~25min, 15% A→30% A; 25~40min, 30% A; 40~41min, 30% A→5% A; 41~45min, 5% A). The detection wavelength was 245 nm, flow rat 1.0mL/min, column tempterature 35℃. In the method, excipients and solvents did not Interfere with the peaks of rupestonic acid and (R, S)-epigoitrin; rupestonic acid and (R, S)-epigoitrin were well separated from adjacent impurity peaks. The calibration curves of rupestonic acid and (R, S)-epigoitrin were in good linearity within the ranges of 24~120 μg /mL (R2 = 0.9992) and 4.8~24 μg /mL (R2 = 0.9997),(The average recovery of rupestonic acid in the three groups of samples were 98.4%, 97.9% and 98.1%, and the RSD were 1.31%,1.02% and 1.33%；The average recovery of (R, S)-epigoitrin in the three groups of samples were 98.8%, 98.3% and 97.5%, and the RSD were 0.56%,0.88% and 1.14%). The results indicated that the method was simple, rapid, accurate and repeatable, and can used to control the content of rupestonic acid and (R, S)-epigoitrin in Haobanqing granules.