Metabolite profiling of human amniotic fluid by hyphenated nuclear magnetic resonance spectroscopy
Metabolite profiling of human amniotic fluid by hyphenated nuclear magnetic resonance spectroscopy
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DOI：
10.1021/ac800907f
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发表时间：
2008-08-01
影响因子：
7.4
通讯作者：
Gil, Ana M.
中科院分区：
文献类型：
--
作者：
Graca, Goncalo;Duarte, Iola F.;Gil, Ana M.
人类羊水(HAF)的代谢谱对于诊断母亲或胎儿的疾病具有潜在的意义。为了建立一个全面的HAF代谢物数据库，首次使用了联用核磁共振对HAF进行系统的分析。实验分别采用反相(RP)和离子交换液相色谱(LC)，以检测较少和较多的极性化合物。在RP-LC条件下，氨基酸、部分糖类和黄嘌呤类物质得到很好的分离。随后的核磁共振和MS分析使30个化合物得以快速鉴定，其中包括我们所知的首次在HAF中鉴定的3-甲基-2-氧戊酸酯和4-氨基-马尿酸。在离子交换LC条件下，检测到30种不同的化合物，包括糖、有机酸、几种有机酸衍生物和氨基酸。在本实验中，首次从HAF中鉴定出5个化合物：D-木糖醇、氨基酸衍生物(N-乙酰丙氨酸、N-乙酰甘氨酸、2-氧代亮氨酸)和异戊酸酯。讨论了某些代谢物(咖啡因、对黄嘌呤、D-木糖醇、山梨醇)的非内源性性质。联用核磁共振可以快速检测HAF中类似的60种代谢物，其中一些由于丰度低(Mu M)和信号重叠而不能被标准核磁共振检测到，从而使得能够为HAF建立一个扩展的代谢物数据库。
The metabolic profiling of human amniotic fluid (HAF) is of potential interest for the diagnosis of disorders in the mother or the fetus. In order to build a comprehensive metabolite database for HAF, hyphenated NMR has been used, for the first,time, for systematic HAF profiling. Experiments were carried out using reverse-phase (RP) and ion-exchange liquid chromatography (LC), in order to detect less and more polar compounds, respectively. RP-LC conditions achieved good separation of amino acids, some sugars, and xanthines. Subsequent NMR and MS analysis enabled the rapid identification of 30 compounds, including 3-methyl-2-oxovalerate and 4-amino-hippurate identified in HAF for the first time, to our knowledge. Under ion-exchange LC conditions, a different set of 30 compounds was detected, including sugars, organic acids, several derivatives of organic acids, and amino acids. In this experiment, five compounds were identified for the first time in HAF: D-Xylitol, amino acid derivatives (N-acetylalanine, N-acetylglycine, 2-oxoleucine), and isovalerate. The nonendogenous nature of some metabolites (caffeine, paraxanthine, D-Xylitol, sorbitol) is discussed. Hyphenated NMR has allowed the rapid detection of similar to 60 metabolites in HAF, some of which are not detectable by standard NMR due to low abundance (mu M) and signal overlap thus enabling an extended metabolite database to be built for HAF.