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Efficient High Performance Liquid chromatograph/ultraviolet Method for Determination of Diclofenac and 4'-hydroxydiclofenac in Rat Serum

Efficient High Performance Liquid chromatograph/ultraviolet Method for Determination of Diclofenac and 4'-hydroxydiclofenac in Rat Serum

Lata Kaphalia, Bhupendra S Kaphalia, Santosh Kumar, Mary F Kanz, Mary Treinen-Moslen

J Chromatogr B Analyt Technol Biomed Life Sci. 2006 Jan 18;830(2):231-7.

PMID: 16301007

Abstract:

A rapid and sensitive high-performance liquid chromatographic method was developed for determination of diclofenac and its major metabolite, 4'-hydroxydiclofenac, in serum from rats treated with diclofenac. The method is simple with a one-step extraction procedure, isocratic HPLC separation, and UV detection at 280 nm. Use of N-phenylanthranilic acid as the internal standard provided good accuracy without interference by endogenous compounds or 5-hydroxydiclofenac, another metabolite of interest. Limits of detection for diclofenac and 4'-hydroxydiclofenac were 0.0225 and 0.0112 microg/ml, respectively. Average extraction efficiencies of diclofenac, 4'-hydroxydiclofenac, and the internal standard were >/=76%. The method was applied to serum collected at 3h after rats were treated with an experimentally useful dosage range of 3, 10 and 50mg/kg diclofenac. Recovery (as a percentage of dose) for the 4'-hydroxy metabolite in serum was found to consistently average from 0.10 to 0.12% following each dosage, whereas recovery of diclofenac in serum declined from 0.45 to 0.37%. Thus, the method is suitable for measurement of a major diclofenac metabolite in experimental studies.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP64118849-B	4′-Hydroxydiclofenac		64118-84-9	Price

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