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 K2 HPO4 ( 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.%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0.5～50 μg· mL-1范围内线性良好,r=0.9995,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.
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