菌体中L-5-甲基四氢叶酸的HPLC法检测  

Determination of L-5-methyltetrahydrofolate in bacteria by HPLC

作　　者：卞筱泓[1] 刘娅梅[1] 许激扬[1] 陈宗梦[1] 

机构地区：[1]中国药科大学生物化学教研室,南京210009

出　　处：《中国新药杂志》2012年第15期1743-1745,共3页Chinese Journal of New Drugs

基　　金：中央高校基本科研业务费专项(JKQ2011046)

摘　　要：目的:建立高效液相色谱法测定菌体中L-5-甲基四氢叶酸(L-5-MTHF)的方法。方法:加热法破碎细胞,强碱性阴离子交换树脂纯化,高效液相色谱柱为C18反向柱,流动相为K2HPO4(0.02 mol.L-1,pH7.2)-甲醇(84∶16)溶液,流速为1.0 mL.min-1,柱温为40℃,检测波长为290 nm。结果:L-5-MTHF在0.5～50μg.mL-1范围内线性良好,r=0.999 5,RSD为1.11%。结论:色谱法灵敏、准确、重现性好,可用于菌体中L-5-甲基四氢叶酸的定性与定量。Objective： To establish an HPLC method for the content determination of L-5-methylenetetra- hydrofolate （L-5-MTHF） in bacteria. Methods： Cells were disrupted by heating, L-5-MTHF was purified by a strong anion-exchange solid-phase extraction. HPLC analysis was performed on C18 column and the mobile phase was K2HPO4 （0.02 mol·L^（-1） , pH 7.2） -methanol （84： 16）. The flow velocity was 1.0 mL·min^（-1） ; the column temperature was 40℃ ; the detection wavelength was 290 nm. Results： The linear range was 0.5 -50 μg·mL^（-1） （r ： 0. 999 5） , and the RSD was 1.11%. Conclusion ： This HPLC method is sensitive and accurate with good repro- ducibility, and can be used in qualitative and quantitative analysis of L-5-MTHF.

关 键 词：L-5-甲基四氢叶酸 高效液相色谱 菌体 

分 类 号：R927.2[医药卫生—药学]