中文摘要：

研究了三磷酸腺苷（ATP）及其代谢物在细胞内的含量以及2-叔丁基-1,4-苯醌（TBBQ）对ATP及其代谢产物在细胞内含量的影响。建立了一种高效液相色谱法（HPLC）用于快速分离、检测细胞内ATP及其代谢产物（二磷酸腺苷（ADP）和一磷酸腺苷（AMP））的含量。使用岛津高效液相系统及艾杰尔Venusil MP C18柱,采用等度洗脱的方式。流动相A相为50 mmol/L磷酸氢二钠和15 mmol/L三甲胺（TEA）,用醋酸（HAc）调节pH至7.88;流动相B相为甲醇。采用建立的高效液相色谱法得到了3种代谢物的工作曲线,相关系数高（R^2≥0.999 6）,MRC-5细胞中3种代谢物的含量均在线性范围（0.1-100μmol/L）内。该方法检出限低。采用预冷的80%（体积分数）甲醇水溶液提取细胞内的代谢物。该研究建立的方法成功地应用于检测MRC-5细胞中的ATP、ADP和AMP的含量,结果表明,TBBQ会对ATP、ADP、AMP在细胞内的含量产生影响,但TBBQ浓度和ATP、ADP以及AMP在MRC-5细胞内浓度的关系比较复杂。

英文摘要：

This study was aimed to provide insight regarding the intracellular metabolites of adenosine 5′-triphosphate（ATP）and whether 2-tert-butyl-1,4-benzoquinone（TBBQ）affects cell metabolites. A rapid high performance liquid chromatography（HPLC）protocol was developed for the separation and quantitation of ATP and its metabolites（adenosine diphosphate（ADP）and adenosine monophosphate（AMP））in cells. Chromatographic separation was performed using a Shimadzu HPLC system equipped with an Agela Venusil MP C18column;isocratic elution was adopted. The mobile phase comprised solvent A（50 mmol / L disodium hydrogen phosphate and 15 mmol / L trimethylamine（TEA）;pH adjusted to 7. 88 using acetic acid（HAc））and solvent B（methanol）. The correlation coefficients of the three analytes were very high（R^2≥0. 999 6）,and the contents of the three metabolites in the MRC-5 cells were within the linear ranges（0. 1- 100 μmol / L）. The limits of detection for the detected three compounds were low. Samples were extracted from cells（after exposure and non-exposure to quinones）using 80%（v / v）methanol aqueous solution. The method developed in this study was successfully applied to detect ATP,ADP and AMP in MRC-5 cells,and the results demonstrated that ATP,ADP,AMP levels in cells were affected by TBBQ,but the relations between the concentration of TBBQ and the level of ATP,ADP and AMP were complex.