RRC ID 67572
著者 Fukuuchi T, Yamaoka N, Kaneko K.
タイトル Analysis of Intra- and Extracellular Levels of Purine Bases, Nucleosides, and Nucleotides in HepG2 Cells by High-performance Liquid Chromatography.
ジャーナル Anal Sci
Abstract To evaluate cellular uptake and purine transport, we developed a high-performance liquid chromatography method for intra- and extracellular purine quantification. Our aim was to develop an effective method for simultaneously quantifying the substrate and metabolites with high sensitivity. C18 columns from different manufacturers were tested for simultaneous quantification of 22 different purine bases, nucleosides, and nucleotides. We used a YMC-Triart C18 column. The analysis conditions, including extraction solutions for the cells and cell culture medium, were optimized to achieve good quantification. Linearity, accuracy, determination limits, and recovery were assessed and showed good performance. The developed HPLC method was successfully applied to the qualitative analysis of 22 different intra- and extracellular purines, demonstrating that it is useful for studying the overall pattern of purine metabolism. This method could also be useful for evaluating metabolic dynamics of purines under a variety of stimulatory conditions of culture cells.
巻・号 31(9)
ページ 895-901
公開日 2015-1-1
DOI 10.2116/analsci.31.895
PMID 26353955
MeSH Chromatography, High Pressure Liquid / methods* Extracellular Space / metabolism* Hep G2 Cells Humans Intracellular Space / metabolism* Nucleosides / metabolism* Nucleotides / metabolism* Purines / metabolism*
IF 2.049
リソース情報
ヒト・動物細胞 Hep G2