Author/Authors :
MADNI, IASAD ULLAH The lslamia University of Bahawalpur - Faculty of Pharmacy and Alternative Medicine - Department of Pharmacy, Pakistan , AHMAD, MAHMOOD Islamia University of Bahawalpur - Faculty of Pharmacy and Alternative Medicine - Department of Pharmacy, Pakistan , AKHTAR, INAYEED lslamia University of Bahawalpur - Faculty of Pharmacy and Alternative Medicine - Department of Pharmacy, Pakistan , ASHRAF, MOHAMMAD lslamia University of Bahawalpur - Faculty of Pharmacy and Alternative Medicine - Department of Pharmacy, Pakistan , SHUJA, ZEBA AHMAD Schazoo Laboratories, Pakistan
Abstract :
A simple and sensitive HPLC method was developed for the determination of ethionamide (ETA) in serum. The method was based on liquid liquid extraction with acetonitrile causing precipitation of serum proteins, drying of supernatant layer and reconstitution with mobile phase. The separation was done by reverse phase chromatography using a hypersil ODS column (250 mm x 4.6 mm, 5µm particle size) and detection at 291 nm. The mobile phase consisted of acetonitrile: phosphate buffer (75:25) which was delivered at 1.5 mL/ min flow rate. The retention time of ETA was found to be 3.8 min. The assay was linear in the concentration range of 0.1-4 µg/ mL. Within day and between day precision extract by relative standard deviation was less than 4 % and inaccuracy did not exceed 3 %. This method is an improved method wherein instead of solid phase extraction, liquid liquid extraction is carried out which resulted in increased sensitivity.
