ANALYTICAL METHOD FOR SEPARATION OF AMINOPHENOL ISOMERS BY REVERSED PHASE CHROMATOGRAPHY

Rajendra Phadke*, Ganesh Hande, Deepesh Patil and Dr. Amit Gosar

ABSTRACT

The purpose of this research study is to develop simple, precise, accurate and economical method for determination and speration of Aminophenol isomers by reserverse phase chromatography. This chromatographic method was developed on Thermo, Hypersil BDS C18, 250 mm x 4.6 mm, 5 μm or equivalent column. The mobile phase used for speperation was 0.01M diammonium hydrogen phosphate buffer adjusted pH 5.5±0.05 with ortho phosphoric acid with secondary mobile phase is methanol. The flow rate of mobile phase was 1.0 ml per minute with intial composition 90 % diammonium hydrogen phosphate buffer and 10% methanol solvent changing to 30% and 70% within 15 minutes then stabalised with initial composition of mobile phase.The detection of isomer of aminophenol was observed at walength 272 nm and peak responcese are very good. The deteceted isomer is about 0.003ppm and further to be quitified at 0.01ppm level concentration. The relative standard deviation of isomer aminophenol was 0.69%. This developed method was validated as per ICH guideline and found out to be linear, accurate, specific, selective, precise, and robust. The correlation coefficient for main aminophenol was 1.000 and the co-isomer re-sponceses where linear from 0.10 ppm to 0.8 ppm. The obtained recovery of isomers were found to be between 95 to 105 percentage and the sample solution was stable up to 12 hrs in dark places hence this method can be success-fully applied for the determination of isomers of Aminophenols.

Keywords: Aminophenol, Isomer, Binary Gradient, HPLC, method development, validation.