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10～500 μg·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。
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.
关键词
莫那克林J /
辛伐他汀 /
洛伐他汀 /
高效液相色谱法 /
生物合成
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Key words
monacolin J /
simvastatin /
lovastatin J /
HPLC /
biosynthesis
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包如胜 孙鹏 杨仲毅 杨勇 王然 李娜.
生物合成过程中莫那克林J的高效液相色谱法测定方法[J]. 中国药学杂志, 2012, 47(7): 543-546
HPLC Analysis of Monacolin J in Biosynthesis[J]. Chinese Pharmaceutical Journal, 2012, 47(7): 543-546
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参考文献
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脚注
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基金
国家“重大新药创制”科技重大专项（2008ZX09204-008）
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