期刊
COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
卷 4, 期 5, 页码 -出版社
ELSEVIER SCIENCE BV
DOI: 10.5936/csbj.201301012
关键词
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资金
- National Science Foundation [MCB-0822581]
- NIH Pre-Doctoral Training Program in Genetics T32 grant [GM008629]
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [T32GM008629] Funding Source: NIH RePORTER
Nicotinamide adenine dinucleotide (NAD(+)) is a coenzyme for hydride transfer reactions and a substrate for sirtuins and other NAD(+)-consuming enzymes. The abundance of NAD(+), NAD(+) biosynthetic intermediates, and related nucleotides reflects the metabolic state of cells and tissues. High performance liquid chromatography (HPLC) followed by ultraviolet-visible (UV-Vis) spectroscopic analysis of NAD(+) metabolites does not offer the specificity and sensitivity necessary for robust quantification of complex samples. Thus, we developed a targeted, quantitative assay of the NAD(+) metabolome with the use of HPLC coupled to mass spectrometry. Here we discuss NAD(+) metabolism as well as the technical challenges required for reliable quantification of the NAD(+) metabolites. The new method incorporates new separations and improves upon a previously published method that suffered from the problem of ionization suppression for particular compounds.
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