High information throughput analysis of nucleotides and their isotopically enriched isotopologues by direct-infusion FTICR-MS.
High information throughput analysis of nucleotides and their isotopically enriched isotopologues by direct-infusion FTICR-MS.
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DOI：
10.1007/s11306-011-0388-y
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发表时间：
2012
期刊：
影响因子：
3.6
通讯作者：
Fan, Teresa W. -M.
中科院分区：
文献类型：
--
作者：
Lorkiewicz, Pawel;Higashi, Richard M.;Lane, Andrew N.;Fan, Teresa W. -M.
关键词：
傅立叶变换离子回旋共振质谱仪(FTICR-MS)能够获得稳定同位素分辨代谢组学(SIRM)无与伦比的高质量同位素数据。这种能力推动了对连续离子引入的需求，以获得最佳的同位素比率。本文报道了采用离子对样品制备方法进行LC-MS分析，用FTICR-MS同时分析了极性粗提物中的单核苷酸和二核苷酸。这包括在含有C18固定相的吸管尖端快速清理提取的核苷酸，这使得直接进样纳米电喷雾FTICR-MS能够以纳摩尔浓度对核苷酸及其13C同位素进行全球分析，只需5分钟的数据采集。分辨率和质量准确度使大多数核苷酸物种的计算机辅助明确归属，包括腺嘌呤、鸟嘌呤、尿嘧啶和胞嘧啶核苷酸的所有磷酸化形式，NAD+，NADH，NADP+，NADPH，环核苷酸，几个UDP-己糖，及其所有13C同位素同系物。该方法被应用于在[U-13C]葡萄糖中生长的人肺腺癌A549细胞的SIRM研究中，有或没有抗癌剂甲基硒酸。在m/z分辨率为400,000的情况下，核苷酸的13C同位素能够从所有其他元素同位素中完全分解，从而能够准确地测定它们的13C部分富集度。该方法实现了用于SIRM研究中代谢途径重建的核苷酸的高样本和高信息吞吐量分析。
Fourier transform-ion cyclotron resonance-mass spectrometry (FTICR-MS) is capable of acquiring unmatched quality of isotopologue data for stable isotope resolved metabolomics (SIRM). This capability drives the need for a continuous ion introduction for obtaining optimal isotope ratios. Here we report the simultaneous analysis of mono and dinucleotides from crude polar extracts by FTICR-MS by adapting an ion-pairing sample preparation method for LC-MS analysis. This involves a rapid cleanup of extracted nucleotides on pipet tips containing a C18 stationary phase, which enabled global analysis of nucleotides and their 13C isotopologues at nanomolar concentrations by direct infusion nanoelectrospray FTICR-MS with 5 minutes of data acquisition. The resolution and mass accuracy enabled computer-assisted unambiguous assignment of most nucleotide species, including all phosphorylated forms of the adenine, guanine, uracil and cytosine nucleotides, NAD+, NADH, NADP+, NADPH, cyclic nucleotides, several UDP-hexoses, and all their 13C isotopologues. The method was applied to a SIRM study on human lung adenocarcinoma A549 cells grown in [U-13C] glucose with or without the anti-cancer agent methylseleninic acid. At m/z resolving power of 400,000, 13C-isotopologues of nucleotides were fully resolved from all other elemental isotopologues, thus allowing their 13C fractional enrichment to be accurately determined. The method achieves both high sample and high information throughput analysis of nucleotides for metabolic pathway reconstruction in SIRM investigations.
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影响因子：
3.6
作者：
Fan, Teresa W-M;Yuan, Peixiong;Lane, Andrew N.;Higashi, Richard M.;Wang, Yun;Hamidi, Anahita B.;Zhou, Rulun;Guitart, Xavier;Chen, Guang;Manji, Husseini K.;Kaddurah-Daouk, Rima
通讯作者：
Kaddurah-Daouk, Rima
影响因子：
3.6
作者：
Fan, Teresa W-M.;Tan, Jinlian;McKinney, Martin M.;Lane, Andrew N.
通讯作者：
Lane, Andrew N.
影响因子：
2
作者：
Mei, H;Hsieh, YS;Korfmacher, WA
通讯作者：
Korfmacher, WA
影响因子：
7.4
作者：
Edwards, JL;Kennedy, RT
通讯作者：
Kennedy, RT
影响因子：
4.3
作者：
Dodbiba, Edra;Breitbach, Zachary S.;Armstrong, Daniel W.
通讯作者：
Armstrong, Daniel W.