ABSTRACT
A stability-indicating reverse phase–high performance liquid chromatography (RP–HPLC) method was developed and validated for the determination of atazanavir sulfate in tablet dosage forms using C18 column Phenomenix (250 mm×4.6 mm, 5μm) with a mobile phase consisting of 900 mL of HPLC grade methanol and 100 mL of water of HPLC grade. The pH was adjusted to 3.55 with acetic acid. The mobile phase was sonicated for 10 min and filtered through a 0.45μm membrane filter at a flow rate of 0.5 mL/min. The detection was carried out at 249 nm and retention time of atazanavir sulfate was found to be 8.323 min. Linearity was observed from 10 to 90μg/mL (coefficient of determination R2 was 0.999) with equation, y=23.427x+37.732. Atazanavir sulfate was subjected to stress conditions including acidic, alkaline, oxidation, photolysis and thermal degradation, and the results showed that it was more sensitive towards acidic degradation. The method was validated as per ICH guidelines.
ABSTRACT
A highly sensitive ultra-fast LC-MS/MS based bioanalytical method for the measurement of Rabeprazole in human plasma was developed and validated using 13C-D3-Rabeprazole as internal standard. Rabeprazole is a sulfabenzimidazole class of compounds, setting chromatography for these classes of compounds is always a challenge. Rabeprazole was extracted from human plasma samples by liquid-liquid extraction (LLE) and separated on a short reverse phase Ascentis® Express C18, 50 mm × 4.6 mm, 2.7 μm column by isocratic elution with 40% 10 mM ammonium acetate solution and 60% acetonitrile at a flow rate of 0.700 mL/min. Rabeprazole and its labeled internal standard were detected by multiple reaction monitoring (MRM) mode using electro spray ionization (ESI). All the validation parameters as per current guidelines like specificity, selectivity, accuracy, precision, recovery, matrix factor, haemolysis effect and stability are assessed in human plasma. Rabeprazole was found to be linear over a range of 0.1 ng/mL to 150 ng/mL in human plasma. LLOQ of0.1 ng/mL is sensitive enough for application to different clinical studies. The intra- and inter-day precision was less than 10% and accuracy was within -3.33 to 10.00%. Recovery (70%) was consistent across the linear dynamic range of the method. The validated method is a simple, accurate, precise and robust to measure Rabeprazole in human plasma and could be used for application to any format of clinical studies.