Analysis of selected sugars and sugar phosphates in mouse heart tissue by reductive amination and liquid chromatography-electrospray ionization mass spectrometry.
Analysis of selected sugars and sugar phosphates in mouse heart tissue by reductive amination and liquid chromatography-electrospray ionization mass spectrometry.
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DOI：
10.1021/ac400769g
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发表时间：
2013-06-18
影响因子：
7.4
通讯作者：
Borchers, Christoph H.
中科院分区：
文献类型：
--
作者：
Han, Jun;Tschernutter, Vera;Yang, Juncong;Eckle, Tobias;Borchers, Christoph H.
Sensitive and reliable analysis of sugars and sugar phosphates in tissues and cells is essential for many biological and cell engineering studies. However, the successful analysis of these endogenous compounds in biological samples by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) is often difficult because of their poor chromatographic retention properties in reversed-phase LC, the complex biological matrices, and the ionization suppression in ESI. This situation is further complicated by the existence of their multiple structural isomers in vivo. This work describes the combination of reductive amination using 3-amino-9-ethylcarbazole, with a new LC approach using a pentafluorophenyl core–shell ultrahigh performance (UP) LC column and methylphosphonic acid as an efficient tail-sweeping reagent for improved chromatographic separation. This new method was used for selected detection and accurate quantitation of the major free and phosphorylated reducing sugars in mouse heart tissue. Among the detected compounds, accurate quantitation of glyceraldehyde, ribose, glucose, glycerylaldehyde-3-phosphate, ribose-5-phosphate, glucose-6-phosphate, and mannose-6-phosphate was achieved by UPLC/multiple-reaction monitoring (MRM)-MS, with analytical accuracies ranging from 87.4% to 109.4% and CVs of ≤8.5% (n = 6). To demonstrate isotope-resolved metabolic profiling, we used UPLC/quadrupole time-of-flight (QTOF)-MS to analyze the isotope distribution patterns of C3 to C6 free and phosphorylated reducing sugars in heart tissues from 13C-labeled wild type and knockout mice. In conclusion, the preanalytical derivatization-LC/ESI-MS method has resulted in selective determination of free and phosphorylated reducing sugars without the interferences from their nonreducing structural isomers in mouse heart tissue, with analytical sensitivities in the femtomole to low picomole range.
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影响因子：
4.4
作者：
Valdes, Jorge;Pedroso, Inti;Quatrini, Raquel;Dodson, Robert J.;Tettelin, Herve;Blake, Robert, II;Eisen, Jonathan A.;Holmes, David S.
通讯作者：
Holmes, David S.
影响因子：
82.9
作者：
通讯作者：
--
影响因子：
3.1
作者：
Wakamatsu, A;Morimoto, K;Kudoh, S
通讯作者：
Kudoh, S
影响因子：
4.1
作者：
Jeong, Ji-Seon;Kwon, Ha-Jeong;Hong, Seon-Pyo
通讯作者：
Hong, Seon-Pyo
影响因子：
2.8
作者：
Harada, Kazuo;Ohyama, Yohko;Fukusaki, Eiichiro
通讯作者：
Fukusaki, Eiichiro