Simple and Sensitive HPLC Method with Fluorescence Detection for the Measurement of Ibuprofen in Rat Plasma: Application to a Long-Lasting Dosage Form

Author： Hassan Ahmed Sheikh Sapin Anne Ubrich Nathalie Maincent Philippe Bolzan Claire Leroy Pierre

Publisher： Informa Healthcare

ISSN： 0363-9045

Source： Drug Development and Industrial Pharmacy, Vol.34, Iss.10, 2008-10, pp. : 1064-1070

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Abstract

A simple and sensitive high-performance liquid chromatography (HPLC) assay applied to the measurement of ibuprofen in rat plasma has been developed. Two parameters have been investigated to improve ibuprofen detectability using fluorescence detection: variation of mobile phase pH and the use of β-cyclodextrin (β-CD). Increasing the pH value from 2.5 to 6.5 and adding 5 mM β-CD enhanced the fluorescence signal (λexc = 224 nm; λem = 290 nm) by 2.5 and 1.3-fold, respectively, when using standards. In the case of plasma samples, only pH variation significantly lowered detection and quantification limits, down to 10 and 35 ng/mL, respectively. Full selectivity was obtained with a single step for plasma treatment, that is, protein precipitation with acidified acetonitrile. The validated method was applied to a pharmacokinetic study of ibuprofen encapsulated in microspheres and subcutaneously administered to rats.