DEVELOPMENT AND VALIDATION OF A STABILITY INDICATING HPLC METHOD FOR THE DETERMINATION OF NILOTINIB HYDROCHLORIDE IN BULK AND PHARMACEUTICAL DOSAGE FORM

Objective: To develop a rapid, accurate, linear, sensitive and stability indicating RP-HPLC method for the determination of nilotinib in bulk and pharmaceutical dosage forms in the presence of its four related substances.Methods: The RP-HPLC method was developed for the chromatographic separation of nilotinib and its impurities by using waters Xterra RP-18 (150*4.6 mm, 3.5 µm) column with a mobile phase combination of 10 mM ammonium formate with pH-3.5 and acetonitrile in gradient mode. An injection volume of 20 µl. Flow rate was 1.0 ml/min and detection was carried a wavelength of 250 nm. The method was validated as per ICH guidelines.Results: The retention time for nilotinib and its four impurities were found to be 4.37, 7.40, 8.96, 10.21 and 10.87 min respectively. The linear regression analysis data for the calibration plots showed the good linear relationship in the concentration range of 0.04-3.0 ppm for the nilotinib impurities. The % recovery of nilotinib impurities was found to be 96.8-99.4% in the linearity range. The detection limit (LOD) values were about 0.014, 0.016, 0.005 and 0.03 ppm respectively and the quantification limit (LOQ) values were 0.042, 0.048, 0.014 and 0.09 ppm respectively. The % degradation at various stress conditions like acid, alkaline, oxidative, thermal and photolytic stress was found to be 8.92, 18.35,5.63, 0.88 and 3.89 respectively.Conclusion: The RP-HPLC method compatible with LC-MS was developed for the analysis of nilotinib and its four impurities. It was validated as per the ICH guidelines and found to be linear, robust, precise, accurate, sensitive, stability indicating and can be used for routine as well as stability analysis of capsule dosage forms as well as for drug substance

Solid phase extractive preconcentration of Co(II) and Mn(II) from environmental and biological samples using 2-hydroxy-acetophenone-3-thiosemicarbazone functionalized Amberlite XAD-2 and determination by ICPAES

628-632A method for solid phase extraction of the trace metals, Co(II) and Mn(II), using Amberlite XAD-2 functionalized with a new chelating ligand, 2-hydroxyacetophenone-3-thiosemicarbazone (Amberlite XAD-2-HAPTSC) has been developed. The optimum experimental conditions for the quantitative sorption of two metals, pH, effect of flow rate, concentration of eluent, sorption capacity, kinetics of sorption and the effect of diverse ions on the sorption of analytes have been investigated. At pH 5.0 the quantitative sorption of two metals onto functionalized resin is observed and complete elution of metals from resin has been achieved with 2.0 mol l⁻¹ nitric acid. The detection limits are 0.82 µg g⁻¹ for Co(II) and 0.64 µg g⁻¹ for Mn(II). The sorption half time (t₁/₂) was found to be 7.5 min for Co(II) and 6.5 min for Mn(II), while the sorption capacities in mg/g are 3.2 for Co(II) and 4.1 for Mn(II). The chelating resin can be reused for more than 20 cycles of sorption-desorption without any significant change in sorption of metal ions (<1.5%). The accuracy of the proposed procedure has been evaluated by comparing the results with ICP-MS. The proposed method has been applied for the determination of trace metals in river water and leafy vegetable samples collected from different sources near Tirupati

Phylogeny of Neobursaridium reshapes the systematics of Paramecium (Oligohymenophorea, Ciliophora)

The subclass Peniculia (Oligohymenophorea, Intramacronucleata) is one of the most known groups of the phylum Ciliophora, being composed by very notorious representatives, such as Paramecium and Frontonia. Nevertheless, phylogenetic relationships among genera within this subclass are still far from being resolved. Moreover, for several members of the group the characterization by molecular markers is still lacking, such as for Wenrichia, Clathrostoma, Paraclathrostoma, Didieria and, to date, also for Neobursaridium. The finding of one strain of Neobursaridium gigas from India led to the first molecular characterization of this uncommonly sampled ciliate. The 18S rDNA sequence and the COI sequence were obtained and used for phylogenetic analyses. Moreover, the partial mitochondrial genome of N. gigas was sequenced, annotated and employed for phylogenomics analysis. To increase the sampling effort for the Paramecium clade, several newly obtained 18S rDNA sequences of parameciids are herein presented. Unexpectedly, the inclusion of N. gigas's molecular data in phylogenetics/phylogenomics analyses did not help to solve the complex evolution relationships inside Peniculia. Conversely, it raised new and intriguing questions about Paramecium phylogeny, since N. gigas clustered inside Paramecium clade as sister species of Paramecium bursaria in all the performed analyses. A critical revision of past and present data led to rename N. gigas as Paramecium gigas (Balech, 1941) comb. nov., and triggered the revision of genus Paramecium, with the proposal of the new subgenus Gigaparamecium subgen. nov. Hypotheses on the evolution of giant morphologies in ciliates are also discussed