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    Ionizable isotopic labeling reagents for relative quantification by mass spectrometry
    10.
    发明授权
    Ionizable isotopic labeling reagents for relative quantification by mass spectrometry 有权
    用于质谱相对定量的可离子化同位素标记试剂

    公开(公告)号：US07982070B2

    公开(公告)日：2011-07-19

    申请号：US11726307

    申请日：2007-03-21

    申请人： Lloyd M. Smith ,  Michael R. Shortreed ,  Brian L. Frey ,  Margaret F. Phillips ,  Joshua J. Coon ,  Shane M. Lamos ,  Casey J. Krusemark ,  Peter J. Belshaw ,  Madhusudan Patel ,  Neil L. Kelleher

    发明人： Lloyd M. Smith ,  Michael R. Shortreed ,  Brian L. Frey ,  Margaret F. Phillips ,  Joshua J. Coon ,  Shane M. Lamos ,  Casey J. Krusemark ,  Peter J. Belshaw ,  Madhusudan Patel ,  Neil L. Kelleher

    IPC分类号： C07C211/10 ,  C07C211/11 ,  G01N33/58

    CPC分类号： C07B59/001 ,  C07B59/00 ,  C07B59/008 ,  G01N30/7233 ,  G01N2458/15 ,  Y10T436/163333 ,  Y10T436/17

    摘要： Relative quantification of metabolites by Electrospray Ionization Mass Spectrometry (ESI-MS) requiring a mechanism for simultaneous analysis of multiple analytes in two or more samples. Labeling reagents that are reactive to particular compound classes and differ only in their isotopic kit facilitating relative quantification and providing tangible evidence for the existence of specific functional groups. Heavy and light isotopic forms of methylacetimidate were synthesized and used as labeling reagents for quantification of amine-containing molecules, such as biological samples. Heavy and light isotopic forms of formaldehyde and cholamine were also synthesized and used independently as labeling reagents for quantification of amine-containing and carboxylic acid-containing molecules, such as found in biological samples. Advantageously, the labeled end-products are positively charged under normal acidic conditions involving conventional Liquid Chromatography Mass Spectrometry (LC/MS) applications. Labeled primary and secondary amine and carboxylic acid end-products also generated higher signals concerning mass-spectra than pre-cursor molecules and improved sensitivity. Improved accuracy concerning relative quantification was achieved by mixing heavy and light labeled Arabidopsis extracts in different ratios. Labeling strategy was further employed to ascertain differences in the amounts of amine-containing metabolites for two strains of Arabidopsis seeds.

    摘要翻译： 通过电喷雾离子化质谱（ESI-MS）对代谢物进行相对定量，其需要同时分析两个或多个样品中多种分析物的机制。 标记对特定化合物类别具有反应性的试剂，仅在其同位素试剂盒中有所不同，便于相对定量，并为特定官能团的存在提供有形证据。 合成了甲基亚酰亚胺酸的重和轻同位素形式，并用作定量含胺分子（如生物样品）的标记试剂。 还合成了重和轻的同位素形式的甲醛和胆胺，并且独立地用作用于定量含胺和含羧酸的分子的标记试剂，例如在生物样品中发现的。 有利地，标记的最终产物在常规的液相色谱质谱（LC / MS）应用的正常酸性条件下带正电荷。 标记的伯胺和仲胺和羧酸终产物也产生了比前体分子更高的质谱信号，提高了灵敏度。 通过以不同比例混合重和轻标记的拟南芥提取物，提高了相对定量的准确性。 进一步使用标记策略来确定两种拟南芥种子的含胺代谢物的量的差异。