Development and Validation of HPLC,TLC and Derivative Spectrophotometric Methods for the Analysis of Ezetimibe in the Presence of Alkaline Induced Degradation Products

Reversed phase‐high performance liquid chromatography (RP‐HPLC), thin layer chromatography (TLC) densitometry and first derivative spectrophotometry (1D) techniques are developed and validated as a stability‐indicating assay of ezetimibe in the presence of alkaline induced degradation products. RP‐HPLC method involves an isocratic elution on a Phenomenex Luna 5μ C₁₈ column using acetonitrile: water: glacial acetic acid (50:50:0.1 v/v/v) as a mobile phase at a flow rate of 1.5 mL/min. and a UV detector at 235 nm. TLC densitometric method is based on the difference in Rf‐values between the intact drug and its degradation products on aluminum‐packed silica gel 60 F₂₅₄ TLC plates as stationary phase with isopropanol: ammonia 33% (9:1 v/v) as a developing mobile phase. On the fluorescent plates, the spots were located by fluorescence quenching and the densitometric analysis was carried out at 250 nm. Derivative spectrophotometry, the zero‐crossing method, ezetimibe was determined using first derivative at 261 nm in the presence of its degradation products. Calibration graphs of the three suggested methods are linear in the concentration ranges 1–10 mcg/mL, 0.1–1 mg/mL and 1–16 mcg/mL with a mean percentage accuracy of 99.05 ± 0.54%, 99.46 ± 0.63% and 99.24 ± 0.82% of bulk powder, respectively. The three proposed methods were successfully applied for the determination of ezetimibe in raw material and pharmaceutical dosage form; the results were statistically analyzed and compared with those obtained by the reported method. Validation parameters were determined for linearity, accuracy and precision; selectivity and robustness and were assessed by applying the standard addition technique.     

Oxidation of o‐phenylenediamine to 2,3‐diaminophenazine in the presence of cubic ferrites MFe2O4 (M = Mn,Co, Ni,Zn) and the application in colorimetric detection of H2O2

Metal ferrites nanocrystallites, MFe₂O₄ (M = Mn, Co, Ni, Zn) were prepared by coprecipitation method and characterized by a combination of physico‐chemical and spectroscopic techniques. MFe₂O₄ nanoparticles having particle size in the range 10–35 nm were tested as catalysts in the oxidation of o‐phenylenediamine (OPD) to 2,3–diaminophenazine (DAP) using hydrogen peroxide as oxidant at room temperature. Kinetic data was collected for the catalytic oxidation of OPD to DAP by monitoring the UV–vis absorbance at 415 nm and fit well to the Michaelis–Menten model yielding kinetic parameters K_m (Michaelis–Menten constant) and V_max (maximum rate of reaction). MnFe₂O₄ nanoparticles provide the highest catalytic activity in the oxidation of OPD to DAP at room temperature. A colorimetric method was developed based on the MnFe₂O₄/OPD system for the detection of H₂O₂ in reaction solution. The method has a detection limit of 30 μM for H₂O₂ and wide linear range.