Fast, efficient capillary electrophoresis method for measuring nucleotide degradation and metabolism

被引:39
作者
Qurishi, R [1 ]
Kaulich, M [1 ]
Müller, CE [1 ]
机构
[1] Univ Bonn, Inst Pharmaceut, Dept Pharmaceut Chem Poppelsdorf, D-53115 Bonn, Germany
关键词
electrokinetic injection; nucleotide metabolism; nucleotides; hexokinase; enzymes; nucleotide phosphates; phosphates;
D O I
10.1016/S0021-9673(02)00095-X
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An easy and fast method for the quantitative analysis of nucleotides by capillary zone electrophoresis was developed. The method employing a neutral-bonded capillary and reversed polarity mode provided a good resolution and a short analysis time of less than 5 min. The samples were injected electrokinetically using -6 kV voltage for 30 s and detected by their UV absorbance at 254 nm. Constant current (-45 muA) was applied, and a phosphate buffer, pH 7.4. was used. The detection limits for ATP, UDP, and UTP ranged between 0.14 and 0.28 muM. This method was required for the investigation of the purity of the commercially available nucleotides used in pharmacological studies, In addition, the analytical method was applied to study the metabolism of nucleotides in a cell line, neuroblastomaXglioma hybrid cells (NG108-15), which is used in pharmacological studies with nucleotides, since it contains purine- and pyrimidine-sensitive nucleotide receptors. Furthermore, we used the new method for monitoring enzymatic studies using the enzyme hexokinase to convert nucleotide triphosphates to diphosphates. (C) 2002 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:275 / 281
页数:7
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