title: Adenosine and dopamine simultaneously determined in urine by reversed-phase HPLC, with on-line measurement of ultraviolet absorbance and electrochemical detection.
abstract: This assay method allows the simultaneous determination of adenosine and dopamine in 0.5 mL of human urine within approximately 35 min, by using isocratic, reversed-phase, high-performance liquid chromatography coupled with an on-line detector for ultraviolet absorbance and electrochemical activity. The analytes were rapidly separated on an affinity column packed with phenylboronate-bonded silica. Mean recoveries for adenosine, 2-chloroadenosine (internal standard), dopamine, and (+/-)-isoproterenol (also an internal standard) were 91%, 99%, 88%, and 104%, respectively. The calibration curves for adenosine and dopamine were linear (r = 0.999, P less than 0.01) over the respective concentration ranges of 0.10 to 2.00 mg/L and 0.05 to 1.00 mg/L. Analytical precisions, assessed by the within-day and the day-to-day CVs, were 4.5% and 6.5% for adenosine, and 5.1% and 6.6% for dopamine, respectively. The mean (+/- SD) concentrations of adenosine (1.27 +/- 0.73 mg/L) and dopamine concentrations (0.38 +/- 0.25 mg/L) measured in urine from 19 healthy subjects agreed well with those previously reported.
authors.affiliation: Division of Adult Nutrition, National Institute of Nutrition, Tokyo, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Echizen
authors.forename: R
authors.initials: R
authors.lastname: Itoh
authors.forename: T
authors.initials: T
authors.lastname: Ishizaki
mesh_terms: D015762:2-Chloroadenosine; D000241:Adenosine / Q000031:analogs & derivatives / Q000652:urine*; D000328:Adult; D001897:Boronic Acids; D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000652:urine*; D005260:Female; D006801:Humans; D006851:Hydrochloric Acid; D007545:Isoproterenol; D008297:Male; D000432:Methanol; D008875:Middle Aged; D011786:Quality Control; D012016:Reference Values
chemical_list: D001897:Boronic Acids; D015762:2-Chloroadenosine; D000241:Adenosine; C010686:benzeneboronic acid; D007545:Isoproterenol; D006851:Hydrochloric Acid; D004298:Dopamine; D000432:Methanol