Determination of flutamide and two major metabolites using HPLC–DAD and HPTLC methods
Research Abstract
Determination of Flutamide and Two Major Metabolites
using HPLC-DAD and HPTLC Methods.
Nada S. Abdelwahb (a)& (b), Heba A. H. Elshemy (c), Nehal F.Farid (a)
Flutamide is a potential antineoplastic drug classified as an anti-androgen. It is a therapy
for men with advanced prostate cancer, administered orally after which it undergoes
extensively first pass metabolism in the liver with the production of several metabolites.
These metabolites are predominantly excreted in urine. One of the important metabolites
in plasma is 4-nitro-3-(trifluoromethyl)phenylamine (Flu-1), while the main metabolite in
urine is 2-amino-5-nitro-4-(trifluoromethyl)phenol (Flu-3).
In this work the two metabolites, Flu-1 and Flu-3, have been synthesized, and then
structural confirmation has been carried out by HNMR analysis. Efforts were exerted to
develop chromatographic methods for resolving Flutamide and its metabolites with the use
of acceptable solvents without affecting the efficiency of the methods. The drug along with
its metabolites were quantitatively analyzed in pure form, human urine, and plasma
samples using two chromatographic methods, HPTLC and HPLC-DAD methods. FDA
guidelines for bio-analytical method validation were followed and USP recommendations
were used for analytical method validation. Interference from excipients has been tested
by application of the methods to pharmaceutical tablets. No significant difference was
found between the proposed methods and the official one when they were statistically
compared at p-value of 0.05 %.
Keywords: Flutamide; metabolites; HPTLC, HPLC; plasma; urine.
Research Keywords
Flutamide; metabolites; HPTLC, HPLC; plasma; urine.