【摘要】 建立了检测细胞中嘌呤核苷酸(ATP、ADP、AMP、NAD+和NADP+)的超高效液相色谱-质谱(UPLC-MS)分析方法。采用KinetexTMHILIC柱(50 mm×4.6 mm,2.1μm)进行分离,对细胞样品的萃取方法、流动相组成及质谱参数进行优化。方法学确证表明:各待测物在1.0~100μmol/L范围内线性关系良好,相关系数(r)均大于0.99,平均回收率为95.7%~101.1%,相对标准偏差为1.2%~6.1%。本方法的灵敏度高、简便快速、重复性好,可用于细胞中能量代谢的研究。更多还原

【Abstract】 An ultra performance liquid chromatography-mass spectrometric( UPLC-MS) method was developed for the simultaneous determination of purine nucleotides( e. g. ATP, ADP, AMP,NAD+and NADP+) in mammalian cell. The HPLC separation was carried out on a KinetexTMHILIC column( 50 mm × 4. 6 mm,2. 1 μm). The experimental parameters,including extraction method,compositions of mobile phase and MS parameters were optimized. The results demonstrated that the standard curves were linear in the range of 1. 0-100 μmol / L,with correlation coefficients above0. 99. The average recoveries were in the range of 95. 7%-101. 1% with RSDs of 1. 2%-6. 1%.The method is sensitive and repeatable,and could be applied in the analysis of intracellular energy metabolism.更多还原