SIMULTANEOUS CHEMOMETRIC AND SPECTROPHOTOMETRIC DETERMINATION OF CO-ADMINISTERED LEVAMISOLE AND TRICLABENDAZOLE IN PHARMACEUTICAL DOSAGE FORM USED IN TREATMENT FASCIOLA-INFECTED SHEEP

The goal of this work is to evaluate chemometric and spectrophotometric approaches. Among these techniques will be dual wavelength, simultaneous equation, and ratio derivative method, as well as classical least-squares (CLS), artificial neural networks (ANN) and genetic algorithm artificial neural networks (GA-ANN) models, for determining the binary combination of levamisole (LEVA) and triclabendazole (TCBZ). The zero spectrum of TCBZ and LEVA is compared, and it is seen that their spectra overlap. This means that the spectrophotometric and chemometric methods can be used to find them. Method I: in dual wavelength the absorbance values of LEVA are identical at 205 and 214 nm; hence, these two wavelengths were used for the detection of TCBZ. For both wavelengths, 217 and 225 nm, the absorbance values of TCBZ are identical; hence, these two wavelengths were chosen for the determination of LEVA. The linearity range of LEVA and TCBZ were 2-14 μg/mL and 2-10 μg/mL, respectively. The LOD were 0.487 µg/mL and 0.436 µg/mL for LEVA and TCBZ, respectively. The LOQ were 1.474 µg/mL and 1.320 µg/mL for LEVA and TCBZ, respectively. Method II: in simultaneous equation method the calibration graphs for the methods were constructed by plotting the absorbance values at 210 nm (λmax of LEVA) and at 222 nm (λmax of TCBZ) versus concentrations in μg/mL. The regression plots were found to be linear over the range of 2-14 µg/mL for LEVA and be linear over the ranges of 2-9 µg/mL for TCBZ. The LOD were 0.353 µg/mL and 0.221 µg/mL, and the LOQ were 1.069 µg/mL and 0.669 µg/mL for LEVA and TCBZ, respectively. Method III: In the ratio derivative method the absorption spectra of the LEVA were divided by the absorption spectrum of TCBZ (6 μg/mL), as a divisor, to get the ratio spectra. While the absorption spectra of the TCBZ were divided by the absorption spectrum of LEVA (14 μg/mL), as a divisor, to get the ratio spectra, the regression plots were found to be linear over the range of 1-9 µg/mL for TCBZ, while 2-14 µg/mL for LEVA. The LOD were 0.542 µg/mL and 0.293 µg/mL, and the LOQ were 1.641 µg/mL and 0.887 µg/mL for LEVA and TCBZ, respectively. Two chemometric assisted spectrophotometric models’ CLS, ANN and GA-ANN were the suitable model for the quantitative analysis of LEVA and TCBZ. These methods were developed for analyzing binary mixtures in pharmaceutical dosage form and bulk powders with high recoveries. Following ICH guidelines, the developed techniques underwent validation. The results show that there were no significant differences when all methods were compared statistically to the method described for TCBZ and LEVA.