e., neuralgia (neuronal pain) and neuron degeneration. Gabapentin is proved to be very effective and well tolerated in the treatment of neuropathic pain. Alpha lipoic acid is an universal antioxidant which prevents oxidative damage of neurons. Methylcobalamin increases myelin sheath formation thereby regenerates neuron. Literature survey reveals many reported methods for the analysis of GBP by ultra-violet (UV),6 and 7 high-performance liquid chromatography (HPLC)8, 9, 10 and 11 and high-performance thin-layer chromatography (HPTLC).12 Various methods have been reported for determination of MCB by UV,13, 14, 15, 16 and 17 HPLC5, 17 and 18

and HPTLC.12 Estimation of ALP by UV,19 and 20 HPLC3, 21 and 22 and GC,21 either individually or in combination with other drugs are reported. To the best of our knowledge, there is no analytical method Bioactive Compound Library datasheet reported for simultaneous determination of ternary mixture containing GBP, MCB and ALP. Therefore, an attempt has been made to develop a simple, accurate, rapid and reproducible ratio spectra derivative spectroscopic method for simultaneous determination

of GBP, MCB and ALP in tablet dosage form and validate it, in accordance with ICH guidelines.23 Pharmaceutical Afatinib molecular weight grade of GBP (Zydus Research Center, Ahmedabad, Gujarat, India), ALP (Centurion Laboratories, Vadodara, Gujarat, India) and MCB (Centurion Laboratories, Vadodara, Gujarat, India) were kindly supplied as gift samples, certified to contain >99% (w/w) on dried basis. Commercially available trigabantin 100 (Sun Pharma, Sikkim) tablets claimed to contain 100 mg Gabapentin, 0.5 mg Methylcobalamin and 100 mg Alpha lipoic acid have been utilized in

the present work. Methanol of Analytical grade was purchased from Merck Chemicals, India and Rankem Chemicals, India. Sartolon Polyamide, 0.20 μm pore size membrane filter, Sartorius AG. 37070 Goettingen, Germany, and 0.45 μm pore size, 47 mm Ø, Sartolon Polyamide, Sartorious AG, Germany. Solutions Rolziracetam containing appropriate concentration of GBP, MCB, ALP and mixture of GBP + MCB + ALP in methanol (glassware’s protected with aluminium foil & keep all glassware below 25 °C) were scanned using UV–visible spectrophotometer in “Spectrum mode” in the range of 800−200 nm and their spectra were stored in computer. Using UV Probe software spectrum of mixture was divided by spectrum of GBP (100 μg/ml) and MCB (0.5 μg/ml); GBP (100 μg/ml) and ALP (100 μg/ml); MCB (0.5 μg/ml) and ALP (100 μg/ml) to get ratio spectrum of ALP, MCB and GBP respectively. Ratio spectra of the drugs were smoothed (Δλ = 10) and Libraries converted to first order derivative spectra (Δλ = 10) using UV Probe software. First order ratio derivative spectra of the drugs were overlaid. From the overlain ratio derivative spectra of GBP, MCB and ALP, 731.10 nm, 768.53 nm and 242.21 nm were selected as suitable analytical wavelengths for analysis of GBP, MCB and ALP respectively ( Fig. 1).