Abstract
A method is described for the quantitative determination of nifurpirinol (NFP) in fish by high performance liquid chromatography. Samples were mixed with anhydrous sodium sulfate and extracted with dichloromethane. The extract was cleaned up by florisil column chromatography. Concentration of the acetone eluate yielded a residue, which was dissolved in methanol containing 2-nitro-p-cresol as an internal standard. NFP was separated on a column packed with LiChrosorb RP-18, 5μm, (4mm×150mm) by using 2.5×10-5v/v% acetic acid in acetonitrile-water (45:55, v/v%) as the mobile phase, with detection at 390nm. The recoveries of NFP added to fish were in the range from 81.8% to 93.9% with an average of 84.8% at levels from 0.1ppm to 0.5ppm. The minimum detectable amount of NFP was 1.25ng. The calibration curve was linear over the range from 2.5ng to 150ng.
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