For Ratio spectrum of GBP, MCB and ALP, spectrum of the mixture was divided by standard spectrum of MCB (0.5 μg/ml) and ALP (100 μg/ml); GBP (100 μg/ml) and ALP (100 μg/ml); and MCB (0.5 μg/ml)

and GBP (100 μg/ml) respectively. Obtained ratio spectra were smoothed (Δλ = 10) and converted to first order derivative spectrum (Δλ = 10, SF = 10 for GBP and MCB; Δλ = 10, SF = 1 for ALP). Amplitude (dA/dλ) of GBP, MCB and ALP were measured at 731.10 nm, 768.53 nm and 242.21 nm Hydroxychloroquine supplier respectively. Concentrations of GBP, MCB and ALP were computed by putting value of their amplitudes in respective standard regression equation obtained from calibration curve. The analysis procedure was repeated six times with tablet formulation. Excellent linearity was obtained for all the three drugs in the range of 100–500 μg/ml for GBP and ALP; and 0.5–2.5 μg/ml MCB. Linearity of GBP, MCB and ALP were shown in Fig. 2, Fig. 3 and Fig. 4 respectively. The correlation coefficients (r2) were found to be greater than 0.998 (n = 6) in all instances. LOD and LOQ were found to be 3.09 μg/ml and 9.37 μg/ml for GBP; 0.03 μg/ml and 0.10 μg/ml for MCB; and 4.79 μg/ml and 14.52 μg/ml for ALP ( Table 1). The proposed method afforded high recoveries for GBP,

MCB and ALP tablets. Results obtained from recovery studies shown in Table 2 indicate that see more this assay procedure can be used for routine quality control analysis of this ternary mixture in tablets. Precision of the analytical method was found to be reliable based on % RSD (<2%) corresponding to the peak areas. The % RSD values were less than 2, for intra-day and inter-day precision. Hence, the method was found to be precise for all the three

drugs. In all deliberately varied conditions for robustness study, the % RSD of GBP, MCB and ALP were found to be well within the acceptable limit (<1.5%) for robustness study ( Table 3). The validated method was used in the analysis of marketed conventional tablet trigabantin 100 with a label claim: 100 mg GBP, 500 μg MCB and 100 mg ALP per tablet. The results for the drugs assay shown in Table 4 indicate a good agreement with the label claims. The spectrum of blank does not show any interference at the detection from of GBP, MCB and ALP as it can be seen from the respective spectra ( Fig. 5). The results of stability study of drugs shown in Table 5. The developed Ratio spectra derivative spectroscopic method is simple, accurate and precise for the simultaneous determination of GBP, MCB and ALP from tablets. It was successfully validated in terms of linearity, range, accuracy, precision, LOD, LOQ and robustness in accordance with ICH Guidelines. Thus, the described method is suitable for routine analysis and quality control of pharmaceutical preparations containing these drugs in combination. All authors have none to declare.