RP-HPLC法同时测定酿酒酵母胞内磷酸腺苷和辅酶Ⅰ

doi:10.3969/j.issn.1673-1689.2012.05.008

首页 > 过刊浏览>2012年第31卷第5期 >492-498. DOI:10.3969/j.issn.1673-1689.2012.05.008

RP-HPLC法同时测定酿酒酵母胞内磷酸腺苷和辅酶Ⅰ
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                        10.3969/j.issn.1673-1689.2012.05.008
                    
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基金项目:国家“十一五”科技支撑计划项目(2008BAI63B06);广东省科技计划工业攻关项目(2009A010700004,2010A010500002)

Simultaneous Determination of Adenosine Phosphate and Coenzyme Ⅰ in Cells of Saccharomyces cerevisiae by RP-HPLC

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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。

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In this manuscript,an RP-HPLC method was developed to determine simultaneously adenosine phosphate and coenzyme I in Saccharomyces cerevisiae,in order to monitor metabolic state.The intracellular metabolites extracted by the method of rubbing with liquid nitrogen were separated on an Agilent Zorbax SB-Aq column(5 μm,4.6 mm×150 mm).A 0.025 mmol/L triethylamine-phosphoric acid buffer solution(pH 6.0) and acetonitrile mixture was used as mobile phase,with a flow rate of 1 mL/min.Detection wavelength was set at 254 nm.Under these conditions,three kinds of adenosine phosphate and two kinds of coenzyme I could be well separated.The recoveries were between 98.30% and 103.42%,and the RSD of the method was less than 3%.The results showed that this method is sensitive,accurate,reproducible and can be applied to determinate the content of intracellular adenosine phosphate and coenzyme I of S.cerevisiae.

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李会品,赵谋明,俞志敏,雷宏杰,赵海锋. RP-HPLC法同时测定酿酒酵母胞内磷酸腺苷和辅酶Ⅰ[J].食品与生物技术学报,2012,31(5):492-498.

LI Hui-pin, ZHAO Mou-ming, YU Zhi-min, LEI Hong-jie, ZHAO Hai-feng. Simultaneous Determination of Adenosine Phosphate and Coenzyme Ⅰ in Cells of Saccharomyces cerevisiae by RP-HPLC[J]. Journal of Food Science and Biotechnology,2012,31(5):492-498.

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