HPLC法测定刺囊毛霉微生物转化甘草次酸的主产物含量

Determination of 7β-hydroxyglycyrrhetinic Acid in Microbial Transformation of Glycyrrhetinic Acid by Mucor spinosus by HPLC

摘要:

采用刺囊毛霉AS3.3450对甘草次酸进行微生物转化, 生成的主产物经分析鉴定是7β-羟基甘草次酸。采用高效液相色谱方法, 以C18-ODS为色谱柱, 甲醇-0.03%三氟乙酸水溶液(梯度洗脱)为流动相, 检测波长为:254 nm, 测得在160 r/min、27°C的转化条件下, 底物加量为130 mg/L, 转化时间为9 d时, 主产物得率最高为712 mg/g甘草次酸, 且在选定色谱条件下7b-羟基甘草次酸的线性范围良好, 平均加样回收率为98.1%, 平均标准偏差(RSD)为2.37%。

Abstract:

The major product of microbial transformation of glycyrrhetinic acid by Mucor spinosus AS3.3450 has been identified as 7β-hydroxyglycyrrhetinic acid on the basis of its spectral data. 7β-hydroxyglycyrrhetinic acid was directly determined by HPLC. Methanol and ultra pure water containing 0.03% trifluoroacetic acid was used as the mobile phase on C18 column, the UV detection wavelength was 254 nm. The maximum transformation rate observed was 712 mg 7β-hydroxyglycyrrhetinic acid/g glycyrrhetinic acid on day 9 under 27°C, 160 r/min and substrate at a concentration of 130 mg/L. Results show that the content of 7β-hydroxyglycyrrhetinic acid indicates a good linearity under these conditions with a average recovery of 98.1% and a RSD of 2.37%.