World Journal of
Pharmaceutical and Life Sciences

METHOD DEVELOPMENT AND VALIDATION OF AFATINIB USING CHIRAL HPLC

C. Purushotham Reddy, K. Ramakrishna* and K.M.V. Narayana Rao*

ABSTRACT

A sensitive and robust chiral high performance liquid chromatography (HPLC) with UV method was developed and validated for the quantification of R-isomer in afatinib as per ICH Q2 guideline. In this method, both R and S-isomers were well-separated on CHIRALPAK-IE column (250 X 4.6 mm X 5?) in an isocratic elution mode with the mobile phase comprising of methanol: methyl tertiary butyl ether: diethyl amine (80:20:0.1; v/v) at a flow rate of 0.7 mL/ min. The column oven temperature was maintained at 20ºC and both afatinib and its enantiomer were monitored at 254 nm by UV detector. The validated method was found to be precise, accurate and linear from the range of LOQ level to 150% with respect to sample concentration and the correlation co-efficient was found to be 0.998. Limit of detection and limit of quantifications were found to be 0.00005 and 0.00015 mg/mL, respectively. The validated method was found to be very sensitive and the recoveries were found to be well within the range from 95.65% to 100.73% for R-isomer. Further, the solution stability was also established and the solutions were found to be stable upto 48hrs.

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