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Abstract

DETERMINATION OF GEMFIBROZIL LEVEL AND ITS STABILITY IN HUMAN PLASMA BY FULLY VALIDATED RAPAID HPLC ANALYSIS

Rajaa Farhan Hussein and Muhammad M. Hammami*

ABSTRACT

We optimized and validated a rapid, simple, and sensitive HPLC assay for gemfibrozil measurement in human plasma and studied the stability of gemfibrozil under various laboratory conditions. After deproteination with acetonitrile, gemfibrozil and ibuprofen (internal standard) were eluted on Symmetry C18, 5 μm steel column at room temperature (RT) with a run time of 8 min. The mobile phase consisted of 0.01 M ammonium phosphate (dibasic) buffer (pH = 5.5) and acetonitrile (45:55, v:v). The analytes were detected using Waters 2475 multi λ fluorescence detector with the excitation and emission wavelength set at 242 and 300 nm, respectively. The response was linear over the range of 0.1–100 μg/ml. Extraction recovery of gemfibrozil and inter-run bias and imprecision were ≥ 89% (mean 95%), ≤ 13%, and ≤ 8.9, respectively. In human plasma, gemfibrozil was stable for at least 24 hours at RT (≥ 97%), 8 weeks at –200C (≥ 95%), and after 3 cycles of freeze at –200C and thaw at RT (≥ 90%). In processed samples, it was stable for at least 24 hours at RT (≥ 93%) and 48 hours at –200C (≥ 100%). Stock solution of gemfibrozil (1 mg/ml in methanol) was stable for at least 48 hours at RT (100%) and 8 weeks at –200C (100%). The assay was successfully employed

Keywords: Gemfibrozil, Ibuprofen, HPLC, Validation, Stability


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