高效液相色谱法和胶束电动毛细管色谱法分析细胞提取物中三磷酸核苷的定量分析：比较研究。

Quantitative analysis of ribonucleotide triphosphates in cell extracts by high-performance liquid chromatography and micellar electrokinetic capillary chromatography: a comparative study.

机构信息

Institute of Microbiology University of Padova, via A. Gabelli 63, 35121 Padova, Italy.

出版信息

Anal Chem. 1994 Sep 15;66(18):2981-4. doi: 10.1021/ac00090a028.

PMID:20213951

Abstract

Two analytical methods have been evaluated for their ability to separate and quantitate the intracellular pool of free ribonucleotide triphosphates in four different cell lines. Micellar electrokinetic capillary chromatography and high performance liquid chromatography were compared in terms of speed, sensitivity, and efficiency of analysis, with the former being 3 times faster (about 10 vs 30 min) and having an average minimun detectable quantity of 50 fmol vs 30 pmol and approximately 950 000 theorical plates/m vs 4500. Different protocols of extraction of nucleotides from whole cells were also evaluated with the two analytical methods. Micellar electrokinetic capillary chromatography proved to be a powerful tool for the reproducible and reliable quantitation of ribonucleotide triphosphates, allowing the fast processing of extremely small volumes of sample.

摘要

已经评估了两种分析方法，以确定它们在四种不同细胞系中分离和定量细胞内游离三磷酸核糖核苷酸池的能力。比较了胶束电动毛细管色谱法和高效液相色谱法的速度、灵敏度和分析效率，前者快 3 倍（约 10 对 30 分钟），平均最小检测量为 50 fmol 对 30 pmol，理论塔板数约为 950000 对 4500。还使用这两种分析方法评估了从全细胞中提取核苷酸的不同方案。胶束电动毛细管色谱法被证明是一种用于可靠和可重现地定量三磷酸核糖核苷酸的强大工具，允许快速处理极少量的样品。