摘要
Adenosine and its corresponding nucleotides adenosine 5′-monophosphate (AMP), adenosine 5′-diphosphate (ADP) and adenosine 5′-triphosphate (ATP) are important biomolecules that provide energy and substrates for various cellular biochemical processes[1]. There have been strong demands for sensitive and reliable analysis of these nucleotides because the determination of their levels in cells may provide valuable information for understanding cellular energy metabolism. Analysis of adenosine nucleotides is a big challenge because these chemicals exist together with high cellular background in samples. Moreover, nucleotides are extremely polar due to the presence of multiple phosphate groups that may interfere with chemical determination. In particular, the nucleotide phosphates are not retained under conventional reversed-phase chromatographic conditions because of their extremely high polarity[2]. The most common approach for quantitative analysis of nucleotide phosphates in cellular extracts is to determine indirectly the corresponding parent nucleosides resulted from enzymatic dephosphorylation after the nucleotides were separated by using an anion-exchange solid phase extraction. The generated nucleosides were analyzed by using liquid chromatography with ultra violet absorption detection (LC/UV), radio-immunoassay and liquid chromatography-mass spectrometry-mass spectrometry (LC-MS-MS)[3-6]. Because nucleotides are not retained under conventional reversed-phase conditions, ion-suppression high performance liquid chromatography (HPLC)[7], capillary electrophoresis[8] and ion-pairing HPLC[9-11] have been employed to circumvent the poor retention problem. The high selectivity of MS and MS/MS techniques provides great separation and detection power, making them attractive alternatives for the trace analysis of nucleotides. The application of negative ion electrospray ionization (ESI)-MS to nucleotide analysis has been reviewed[12,13]. The use of negative ion ESI-MS seemed to be a logical starting point for nuc
出处
《色谱》
CAS
CSCD
北大核心
2004年第4期358-360,共3页
Chinese Journal of Chromatography
关键词
离子对液相色谱
飞行时间质谱
联用技术
同时分离
核苷
核苷酸
腺苷
ion-pairing liquid chromatography
quadrupole-time of flight mass spectrometry
adenosine
(nucleotides)
