Premium
Analysis of adenosine by RP‐HPLC method and its application to the study of adenosine kinase kinetics
Author(s) -
Marin Rodrigo Miguel,
Franchini Kleber Gomes,
Rocco Silvana Aparecida
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700194
Subject(s) - high performance liquid chromatography , chromatography , adenosine , chemistry , kinetics , linear range , quantitative analysis (chemistry) , detection limit , analytical chemistry (journal) , biochemistry , physics , quantum mechanics
An RP‐HPLC method for the analysis of adenosine (ADO) has been developed and validated. In the present study, we report an RP‐HPLC‐based method with modifications of mobile phase and shorter retention time that substantially improved the efficiency of ADO analysis. The HPLC separation of the ADO was achieved on a C18 column, using a mobile phase consisting of water, containing 7% v/v ACN, at a flow rate of 0.8 mL/min. The column effluent was monitored by UV detection at 260 nm. A linear response was achieved over the concentration range of 0.25–100.00 μmol/L. The analytical method inter‐ and intra‐run accuracy and precision were better than ± 15%. The LOQ was 0.25 μmol/L, with ADO detection in the range of 6.25 pmol per sample. The method has been applied to the study of adenosine kinase (AK) kinetics.