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目的通过RP-HPLC法检测25种真菌对齐墩果酸的转化情况,并对有转化的真菌菌种进行转化率的测定。方法采用Kromasil C18柱(250 mm×4.6 mm,5μm),流动相为甲醇-体积分数0.1%乙酸水溶液(体积比94∶6),柱温为室温,流速1.0 mL.min-1,检测波长210 nm。结果齐墩果酸质量浓度在10~80 mg.L-1内与峰面积呈良好线性关系,回归方程为A=6.778×106ρ-2.539×104(r=0.999 8)。经HPLC检测25种真菌中有5种真菌对齐墩果酸有转化,测得的转化率分别为56.2%、69.7%、77.9%、81.6%、83.0%。结论该法简便、准确、重现性好,适用于齐墩果酸转化的筛选及转化率的测定。
OBJECTIVE To determine the transformation of oleanolic acid by 25 kinds of fungi by RP-HPLC and to determine the conversion rate of transformed fungi. Methods A Kromasil C18 column (250 mm × 4.6 mm, 5 μm) was used. The mobile phase consisted of methanol and 0.1% acetic acid in volume ratio (94:6). The column temperature was at room temperature and the flow rate was 1.0 mL · min- nm. Results The oleanolic acid showed a good linear relationship with the peak area in the range of 10-80 mg · L-1, the regression equation was A = 6.778 × 106ρ-2.539 × 104 (r = 0.999 8). The results of HPLC showed that oleanolic acid was converted into 5 fungi among 25 fungi. The conversion rates were 56.2%, 69.7%, 77.9%, 81.6% and 83.0%, respectively. Conclusion The method is simple, accurate and reproducible. It is suitable for the selection of oleanolic acid and the determination of conversion rate.