Abstract
A sensitive, reliable and reproducible ion-pair chromatographic (IPC) method with enhanced fluorescent detection suitable for therapeutic drug monitoring was developed and validated for the determination of α1-blockers. The chromatographic and fluorescence enhancement conditions were optimized for maximum sensitivity and selectivity. Sodium dodecyl sulfate (SDS) has a dual role in the newly developed IPC method as an ion pair reagent as well as enhancement of the native fluorescence of the studied drugs. Additionally, a simple isocratic separation mode with mobile phase containing acetonitrile and 20mM phosphate buffer (pH6.3) (60:40, v/v) containing 25mM SDS was the best for separation. The flow rate was set at 0.8mLmin−1 and the effluent was monitored by a fluorescence detector and the wavelengths at emission wavelength of 389nm after excitation at =250nm. The developed IPC method provided rapid separation of α1-blockers with good peak resolution compared with conventional reversed phase (RP-C18) methods. The method was validated in accordance with the requirements of USP-FDA guidelines and shown to be suitable for intended applications. The linearity range for the developed IPC method in rabbit plasma obtained was 0.5–20ngmL−1 with correlation coefficients of more than 0.9985. The limits of detection (S/N=3) and quantification (S/N=10) were 0.16–0.71ngmL−1 and 0.53–2.14ngmL−1, respectively. The inter-day and intra-day precision % RSD was 1.14–3.91 and accuracy of 9.06–102.3%. The proposed IPC–FL method was successfully applied for the bioequivalence study of two formulations for terazosin from different brands in rabbits by a randomized, two-way, single-dose, crossover study and proved to be bioequivalent.
•A simple, reliable, and sensitive IPC-FL method was developed for the determination of α1-blockers in plasma.•SDS has a dual role as an ion pair reagent and enhancement of the native fluorescence of α1-blockers.•IPC improved peak resolution in short analysis time and augmented method selectivity.•IPC-FL method was 1000 times more sensitive than HPLC-UV method.•IPC-FL method was applied successfully to study bioequivalence of TER.