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Journal of Pharmaceutical and Applied Chemistry
An International Journal


Volumes > Vol. 6 > No. 3


New Method Development for Pregabalin Detection in Human Plasma by HPLC-DAD

PP: 61-66
Rania A. Radwan, Nour F. Abouzied, Fawzy A. Badway, Hammad H. A., Ali M. Hassan,
Pregabalin, (S)-3-(amino ethyl)-5-methylhexanoic acid, is one of prescribed drugs with increasing abuse. Pregsal, ((S,E)-3- (((2-hydroxybenzylidene)amino)methyl)-5- methylhexanoic acid) is a novel pregabalin functionalized salicylaldehyde derivative. Present study aimed to development of new method for accurate detection of pregabalin in human plasma by HPLC-DAD through increasing its chromatographic sensitivity by derivatization and application of this method on human volunteers. Separation of pregabalin derivative was done using HPLC-DAD, Agilent 1200 series, on reversed phase columns by phosphate buffer and acetonitrile. Pregabalin in the sample was drevatized using Salicylaldehyde solution with formation of pregsal. Pregsal was extracted using liquid liquid extraction, Pregsal was best extracted using dichloromethane in slightly acidic PH. The best chromatographic separation of pregsal was on Zorbax - C8 (250 mm 4.6 mm, 5 μm) column, gradient mobile phase (phosphate buffer with triethylamine 4.5 and acetonitrile) at 0 minute 65:35and 6.1 minute 50:50 and with flow rate 1.5. Retention time for pregsal 14.10.3 minute and for ibuprofen 17.8 0.3 minute. LOD of pregabalin in extracted samples was 1000 ng/ml and LOQ was 2500 ng/ml. Linearity range was from 2500 to 40000 ng/ml and r2 was 0.988. The mean recovery of pregabalin from plasma was 73.8%. further method validation is needed for the present method.

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