Abstract

RAPID DETERMINATION OF HYDROXYZINE CONCENTRATION IN HUMAN PLASMA BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

Reem Alswayeh, Syed N. Alvi, and Muhammad M. Hammami*

ABSTRACT

A simple, precise, and rapid high performance liquid chromatography (HPLC) method for the determination of hydroxyzine level in human plasma using omeprazole as an internal standard (IS) was developed and validated. Plasma samples containing hydroxyzine were spiked with the IS; 3 ml of tert. Butyl methyl ether were added; and mixture was vortexed for 1 min and then centrifuged for 15 min at 4200 rpm. The organic layer was transferred to a clean tube and dried under and a gentle stream of nitrogen, and the residue was reconstituted in 200 μl mobile phase. The compounds of interest were efficiently separated on 4.6 x 150 mm, Atlantis dc18, 5-μm, steel column, maintaining temperature at 30°C, and were detected with a photodiode array detector set at 230 nm. The mobile phase consisted of 0.05 M Ammonium Acetate buffer (pH ꞊ 4.0, adjusted with phosphoric acid), acetonitrile, and methanol (50:35:15, v:v:v) and was delivered at a flow rate of 1.0 ml/min. No interference in blank plasma or by commonly used drugs was observed, and the detection limit of hydroxyzine was 0.015 μg/ml. The relationship between hydroxyzine concentration in plasma and peak area ratio of hydroxyzine /IS was linear (R2≥0.986) in the range of 0.04 – 0.6 μg/ml. Intra- and inter-day coefficient of variations (CV) and biases were ≤10.9% and ≤11.3%, and ≤10.0 and ≤5.0 respectively. Mean extraction recovery of hydroxyzine and the IS from the plasma samples was ≥86%, Using the method, hydroxyzine was found to be stable under conditions generally encountered in the clinical laboratory (≥89% and ≥92% in processed and unprocessed samples, respectively).Further, the method was successfully employed to measure hydroxyzine level in plasma samples from a healthy volunteer.

Keywords: Hydroxyzine, Omeprazole, Human plasma, HPLC.