通过氢质子（¹H）和磷-31（³¹P）核磁共振波谱法直接同时定量测定三磷酸腺苷（ATP）、二磷酸腺苷（ADP）和一磷酸腺苷（AMP）。

Direct and simultaneous quantification of ATP, ADP and AMP by (1)H and (31)P Nuclear Magnetic Resonance spectroscopy.

作者信息

Lian Yakun, Jiang Hua, Feng Jinzhou, Wang Xiaoyan, Hou Xiandeng, Deng Pengchi

机构信息

Analytical & Testing Center, Sichuan University, Chengdu 610064, China.

Metabolomics and Multidisciplinary Laboratory for Trauma Research, Sichuan Provincial People's Hospital, Sichuan Academy of Medical Sciences, Chengdu 610064, China.

出版信息

Talanta. 2016 Apr 1;150:485-92. doi: 10.1016/j.talanta.2015.12.051. Epub 2015 Dec 20.

DOI:10.1016/j.talanta.2015.12.051

PMID:26838434

Abstract

ATP, ADP and AMP are energy substances with vital biological significance. Based on the structural differences, a simple, rapid and comprehensive method has been established by (1)H and (31)P Nuclear Magnetic Resonance ((1)H-NMR and (31)P-NMR) spectroscopies. Sodium 3-(trimethylsilyl) propionate-2,2,3,3-d4 (TMSP) and anhydrous disodium hydrogen phosphate (Na2HPO4) were selected as internal standards for (1)H-NMR and (31)P-NMR, respectively. Those three compounds and corresponding internal standards can be easily distinguished both by (1)H-NMR and (31)P-NMR. In addition, they all have perfect linearity in a certain range: 0.1-100mM for (1)H-NMR and 1-75 mM for (31)P-NMR. To validate the precision of this method, mixed samples of different concentrations were measured. Recovery experiments were conducted in serum (91-113% by (1)H-NMR and 89-113% by (31)P-NMR).

摘要

ATP、ADP和AMP是具有重要生物学意义的能量物质。基于结构差异，通过氢-1（¹H）和磷-31（³¹P）核磁共振（¹H-NMR和³¹P-NMR）光谱法建立了一种简单、快速且全面的方法。分别选用3-（三甲基硅基）丙酸-2,2,3,3-d4（TMSP）钠和无水磷酸氢二钠（Na₂HPO₄）作为¹H-NMR和³¹P-NMR的内标。这三种化合物及其相应的内标在¹H-NMR和³¹P-NMR中都很容易区分。此外，它们在一定范围内都具有良好的线性：¹H-NMR为0.1-100mM，³¹P-NMR为1-75mM。为验证该方法的精密度，对不同浓度的混合样品进行了测量。在血清中进行了回收率实验（¹H-NMR为91-113%，³¹P-NMR为89-113%）。