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为准确监控酿酒酵母胞内磷酸腺苷和辅酶I的代谢情况,作者建立了酿酒酵母胞内磷酸腺苷和辅酶I的高效液相色谱分析方法。胞内代谢物经液氮研磨提取后,采用Agilent ZorbaxSB-Aq色谱柱(5μm,4.6mm×150mm)分离,流动相0.025mmol/L三乙胺-磷酸缓冲液(pH6.0)与乙腈梯度洗脱,流速1mL/mim,检测波长254nm。在此条件下,3种磷酸腺苷和两种辅酶I达到了较好的分离效果。回收率在98.30%～103.42%之间,RSD<3.00%。结果表明,本法灵敏、准确、重现性好,可用于酿酒酵母胞内磷酸腺苷和辅酶I摩尔质量浓度的测定。
In order to accurately monitor the metabolism of intracellular ATPase and Coenzyme I in Saccharomyces cerevisiae, the authors established a high performance liquid chromatographic method for the determination of intracellular ATP and Coenzyme I in Saccharomyces cerevisiae. The intracellular metabolites were extracted by liquid nitrogen and separated on an Agilent Zorbax SB-Aq column (5 μm, 4.6 mm × 150 mm). The mobile phase was 0.025 mmol / L triethylamine-phosphate buffer (pH 6.0) Off, flow rate 1mL / mim, detection wavelength 254nm. Under these conditions, three kinds of adenosine triphosphate and two coenzyme I achieved a better separation effect. The recoveries ranged from 98.30% to 103.42% with a RSD <3.00%. The results showed that this method was sensitive, accurate and reproducible, and could be used to determine the molar concentration of ATP and CoI in Saccharomyces cerevisiae.