Herein, a Rayleigh light-scattering (RLS) detection method combined with high performance liquid chromatograph (HPLC) without any post-column probe was developed for the separation and determination of three α 1 -adrenoceptor antagonists. The quantitative analysis is benefiting from RLS signal enhancement upon addition of methanol which induced molecular aggregation to form an hydrophobic interface between aggregates and water that produce a sort of superficial enhanced scattering effect. A good chromatographic separation among the compounds was achieved using a Gemini 5u C 18 reversed phase column (250mm×4.6mm; 4μm) with a mobile phase consisting of methanol and ammonium acetate–formic acid buffer solution (25mM; pH=3.0) at the flow rate of 0.7mLmin −1 . The RLS signal was monitored at λ ex =λ em =354nm. A limit of detection (LOD) of 0.065–0.70μgL −1 was reached and a linear range was found between peak height and concentration in the range of 0.75–15μgL −1 for doxazosin mesylate (DOX), 0.075–3.0μgL −1 for prazosin hydrochloride (PRH), and 0.25–5μgL −1 for terazosin hydrochloride (TEH), with linear regression coefficients all above 0.999. Recoveries from spiked urine samples were 88.4–99.0% which is within acceptable limits. The proposed method is convenient, reliable and sensitive which has been used successfully in human urine samples.