Pharmacokinetic study of daclatasivir in rabbit plasma using an eco-friendly factorial-designed spectrofluorimetric method: Greenness assessment

Abstract

A green and simple spectrofluorometric method has been developed for the detection of daclatasivir dihydrochloride in rabbit plasma. The optimum conditions for the plasma preparation procedure were determined using the factorial design technique. The factorial design technique is considered the greenest technique for achieving optimum conditions in a short time. Daclatasivir dihydrochloride was efficiently recovered through a single-step protein precipitation using acetonitrile. The extraction efficiencies were 95.07–97.05 %. The supernatant was scanned at a fixed excitation wavelength of 316 nm. The fluorescence intensity of the emission peak maximum was obtained at 384 ± 2 nm. The proposed method was validated in accordance with the bioanalytical method validation of ICH's and EMA's guidelines. The calibration curve was linear over 0.10–3.00 μg mL-1 (coefficient of determination (R2) = 0.9990), with a lower limit of quantification (LLOQ) at 0.10 μg mL-1. The mean recovery was 101.45%, with a standard deviation of 5.52%. The percentage coefficient of variation of within-run and between-run precision was less than 4.39%. The rabbit plasma pharmacokinetic parameters of two daclatasivir dihydrochloride doses were compared and estimated by the linear-log trapezoidal method using the PK solver application. The results show a significant difference between the two doses concerning AUC(0→∞), AUMC(0→∞), and Cmax, while both Tmax and MRT show non-significant differences. Their elimination rate constants (Ke) and half-lives (t1/2) are almost identical. Finally, the proposed method is excellently green, according to Eco-Score (81) and GAPI. The proposed method is suitable for preclinical trials of new DAC tablet formulations.