超高效液相色谱-质谱法测定细胞中嘌呤核苷酸的方法探究  

Study on Determination of Purine Nucleotides by Ultra Performance Liquid Chromatography- Mass Spectrometry

作　　者：姜丹丹[1] 李伟[1] 周怀彬[1] 张婷[1] 许芳[1] 武佳[1] 吕建新[1] 

机构地区：[1]温州医学院检验医学院&生命科学学院检验医学教育部重点实验室,浙江温州325035

出　　处：《分析测试学报》2013年第10期1202-1206,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金(30900811);浙江省自然科学基金(Y2090753)

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（UP-LC-MS）分析方法.采用KinetexTM HILIC柱（50 mm ×4.6 mm,2.1 μm）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 ～ 100 μmol/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.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 KinetexTM HILIC 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 above 0.99.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.

关 键 词：超高效液相色谱-质谱法 细胞 ATP ADP AMP NAD+ NADP+ 

分 类 号：O657.63[理学—分析化学] Q524[理学—化学]