【摘要】 目的建立辛伐他汀生物合成过程中的莫那克林J的HPLC测定方法。方法色谱柱填料为C18多孔微粒键合硅胶,流动相为0.1%磷酸-乙腈(30∶70),流速为1 mL.min-1,检测波长238 nm;用HPLC控制洛伐他汀发酵液经碱裂解制备的莫那克林J及土曲霉基因改良工程菌直接发酵得到的莫那克林J含量。结果开环莫那克林J在10～500μg.mL-1内线性良好,r为0.999 8;经HPLC检测,洛伐他汀发酵液经碱裂解后提取得到莫那克林J开环酸的含量为92%,收率为96.24%,而土曲霉基因改良工程菌发酵液中则残留较多的洛伐他汀,与莫那克林J比例约为1.1∶1.0。结论本实验建立的方法可以将莫那克林J、辛伐他汀及洛伐他汀的开环形式与闭环形式完全分开,适合用于辛伐他汀生物合成过程莫那克林J的研究。更多还原

【Abstract】 OBJECTIVE To establish an HPLC analysis method of monacolin J(MJ).METHODS MJ sample was analyzed with Hypersil C18 HPLC column.The mobile phase was composed of 0.1% H3PO4 and acetonitrile(30∶70) with flow rate of 1.0 mL·min-1,and the detection wavelength was 238 nm.Monacolin J samples produced from lovastatin fermentation mycelia and engineered Aspergilus terreus were analyzed.RESULTS The calibration curve of monacolin J was linear in the range of 10-500 μg·mL-1 with r of 0.999 8.The purity of crystaline monacolin J from lovastatin fermentation mycelia was 92%,and the purification yield was 96.24%.Meanwhile the engineered Aspergilus terreus produced lovastatin and monacolin J at the ratio of about 1.1∶1.0.CONCLUSION This HPLC method can separate lactone form and acid form of monacolin J,lovastatin and simvastatin very well.This HPLC method can be effectively used in monacolin J research of simvastatin biosynthesis.更多还原