HPLC同时测定赤芝中4种三萜酸的含量

Simultaneous HPLC determination of four triterpenoid acids in Ganoderma lucidum

目的：建立HPLC同时测定赤芝药材中灵芝酸C2、灵芝烯酸A、灵芝酸A和灵芝酸D含量的方法。方法：采用Kromasil C18色谱柱（4.6 mm×250 mm，5 μm）；流动相为乙腈-0.03%磷酸水溶液采用梯度洗脱，流速1.0 mL·min-1，检测波长252 nm，柱温35 ℃。结果：灵芝酸C2、灵芝烯酸A、灵芝酸A和灵芝酸D的线性范围分别为：5.0～50.0 mg·L-1（r=0.999 9），7.2～72 mg·L-1（r=0.999 9），11.67～116.7 mg·L-1（r=0.999 9）和5.32～53.2 mg·L-1（r=0.999 8）；平均回收率（n=3）分别为98.8%（RSD1.5%），99.1%（RSD1.9%），99.5%（RSD1.4%）和98.5%（RSD1.9%）。结论：该方法简便、准确，重复性好，为赤芝药材的质量控制提供了实验依据。

Objective： To determine 4 kinds of triterpenoid acids in Ganoderma lucidum, namely ganoderic acid C2, ganoderenic acid A, ganoderic acid A and ganoderic acid D quantitively. Method： The RP-HPLC method was applied and the separation was performed on a Kromasil C18 analytical column （4.6 mm×250 mm, 5 μm ）. The mobile phase was acetonitrile （A）-water containing 0.03%H3PO4 （B） with gradient elution mode at the flow rate of 1.0 mL·min-1. The detection was set at 252 nm, and the column temperature was 35 ℃. Result： The linear ranges of ganoderic acid C2, ganoderenic acid A, ganoderic acid A and ganoderic acid D were 5.0-50.0 μg·mL-1 （r=0.999 9）, 7.2-72 mg·L-1 （r=0.999 8）, 11.67-116.7 mg·L-1 （r=0.999 9）, 5.32-53.2 mg·L-1 （r=0.999 6）, respectively. The average recoveries （n=9） were 98.8% （RSD 1.5%）, 99.1% （RSD 1.9%）, 99.5% （RSD 1.4%）, 98.5% （RSD 1.9%）, respectively. Conclusion： The method is simple and accurate with a good reproducibility and can be used as a quality control method for G. lucidum of different sources.