Abstract
A specific high-performance thin-layer chromatography (HPTLC) method with fluorescence detection was developed for determination of selective alpha(1)-blockers, including terazosin (TER), doxazosin, prazosin, and alfuzosin. So far, this is the first HPTLC method using fluorescence detection mode for the determination of alpha(1)-blockers. It was designated to combine the specificity attributed to the drug's native fluorescence, the high sample throughputs of the HPTLC methods, and the use of environmentally safe reagents. Good resolution was obtained using simple mobile phase composed of acetonitrile and phosphate buffer (0.02mole/L) adjusted to pH 3.0 (30:70, v/v). The method used fluorescence detection mode and was validated according to the International Conference on Harmonization guidelines. The emission intensity was measured using optical filter K400 after excitation at 332nm. The correlation coefficient in the range 0.9984-0.9997 was obtained in the concentration range 5-100ng/band. The limits of detection and quantification were 3.52-6.91 and 11.76-23.02ng/band, respectively. Forced degradation stability for TER was studied using the developed HPTLC method. The proposed method was successfully applied for the determination of the studied alpha(1)-blockers in pharmaceutical dosage forms and in human plasma without the need for complicated extraction and cleanup steps.