雅致小克银汉霉对莫诺苷生物转化工艺的研究

Microbial transformation of morroniside by Cunninghamella elegans

目的建立UPLC-MS/MS法测定莫诺苷含量,并优选出雅致小克银汉霉对莫诺苷的生物转化工艺条件。方法采用UPLC-MS/MS法测定莫诺苷的含量,以莫诺苷的生物转化率为指标,进行培养基初始pH、底物浓度、转化时间、振摇频率等条件的考察,选出最佳生物转化条件。结果莫诺苷在76.544~19136 ng·mL^-1与峰面积线性关系良好,回归方程为y=0.0003x-0.0286(r=0.9996),方法符合生物样品分析方法的要求。雅致小克银汉霉对莫诺苷最佳的生物转化条件为底物浓度11.48μg·mL^-1、转化时间8 d、摇床振摇频率160 r·min^-1,放置于28℃摇床培养,转化率约90%。结论雅致小克银汉霉的微生物代谢模型对莫诺苷有较高的生物转化率,所建立的莫诺苷UPLC-MS/MS含量测定方法准确有效,能够为莫诺苷的体内体外生物转化研究提供依据。

Objective To determine the content of morroniside by UPLC-MS/MS to optimal bioconversion process condition of morroniside by Cunninghamella elegans.Methods UPLC-MS/MS was used to determine the bioconversion rate of morroniside with Cunninghamella elegans.For the biological conversion,we optimized medium initial pH,concentration of substrate,conversion time and shaking frequency,and selected the optimal bioconversion conditions.Results Morroniside had good linearity at 76.544 ng·mL^-1 to 19136 ng·mL^-1.The regression equation was y=0.0003x-0.0286(r=0.9996).The method conformed to the requirements of the biological sample analysis.The optimum bioconversion conditions of morroniside by Cunninghamella elegans were as follows:the substrate concentration at 11.48μg·mL^-1,conversion for 8 d,shaking frequency at 160 r·min^-1,and culture in a shaking table at 28℃.The bioconversion rate was 90%.Conclusion The microbial metabolism model of Cunninghamella elegans is highly efficient for the bioconversion of morroniside,which helps simplify the in vitro and in vivo drug metabolism.