ANALYSIS OF DICLOFENAC AND 4 OF ITS METABOLITES IN HUMAN URINE BY HPLC

Citation
Rj. Sawchuk et al., ANALYSIS OF DICLOFENAC AND 4 OF ITS METABOLITES IN HUMAN URINE BY HPLC, Pharmaceutical research, 12(5), 1995, pp. 756-762
Citations number
5
Categorie Soggetti
Pharmacology & Pharmacy",Chemistry
Journal title
ISSN journal
07248741
Volume
12
Issue
5
Year of publication
1995
Pages
756 - 762
Database
ISI
SICI code
0724-8741(1995)12:5<756:AODA4O>2.0.ZU;2-S
Abstract
An HPLC method for the determination of diclofenac (DCF) and four of i ts metabolites (3'-hydroxydiclofenac, 4'-hydroxydiclofenac, 5-hydroxyd iclofenac, and 3'-hydroxy-4'-methoxydiclofenac) in human urine is desc ribed. Following base hydrolysis, the samples were neutralized and ext racted. Evaporated extracts were reconstituted in mobile phase contain ing ascorbic acid, and chromatographed, using flow-rate programming, o n a reversed-phase column. Absolute recovery (average), was at least 7 8% for diclofenac and ranged from 75 to 85% for the four metabolites. Standard curves showed linearity over the range of concentrations of 0 .2 to 40 ug/mL, using 0.25 mt of urine. Specificity was demonstrated b y examining chromatograms of extracts of blank urine from 8 volunteers and 24 study subjects. Good accuracy was observed for all compounds o ver the concentration range of 0.2 to 40 ug/mL using 0.25 mt of urine. Based on accuracy and precision criteria, the limit of quantitation f or ah 5 analytes was 0.4 ug/mL, using 0.25 mt of urine. Analysis of ur ine from subjects with normal and reduced renal function who received diclofenac orally demonstrated that total diclofenac and metabolites e xcreted in the urine represented approximately 31% and 4% of an oral d ose of diclofenac, respectively.