title: Analysis of adenosine by RP-HPLC method and its application to the study of adenosine kinase kinetics.
abstract: 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 micromol/L. The analytical method inter- and intra-run accuracy and precision were better than +/- 15%. The LOQ was 0.25 micromol/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.
authors.affiliation: Department of Internal Medicine, School of Medicine, UNICAMP, Campinas, SP, Brazil.
authors.forename: Rodrigo Miguel
authors.initials: RM
authors.lastname: Marin
authors.forename: Kleber Gomes
authors.initials: KG
authors.lastname: Franchini
authors.forename: Silvana Aparecida
authors.initials: SA
authors.lastname: Rocco
mesh_terms: D000241:Adenosine / Q000032:analysis*; D000248:Adenosine Kinase / Q000378:metabolism*; D000818:Animals; D002623:Chemistry Techniques, Analytical / Q000379:methods*; D002845:Chromatography / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D007700:Kinetics; D051379:Mice; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013056:Spectrophotometry, Ultraviolet; D013997:Time Factors; D014867:Water / Q000737:chemistry*
chemical_list: D014867:Water; D000248:Adenosine Kinase; D000241:Adenosine