RP HPLC

A new simple and sensitive RP-HPLC method was developed and validated as per the ICH guidelines for the estimation of fenoverine in bulk and pharmaceutical dosage form. The chromatographic separation was achieved  on Enable 18H C18  column (250 x 4.6mm, 5µm)  with a mobile phase containing acetonitrile and phosphate buffer pH 7 (55:45) at flow rate of 0.8ml/min using in isocratic elution mode. Detection was carried out at 262nm with the retention time of 4.7mins. Linearity in the calibration plot was achieved over the concentration range of 5-25ng/ml with an r2 value of 0.997. The method was validated for accuracy, precision, specificity and selectivity, robustness, detection and quantification limits and system suitability parameters according to ICH guidelines Q2 R1. The detection limit and quantitation limit were found to be 1.3ng/ml and 4ng/ml respectively. Further the validated method was successfully applied for the analysis of fenoverine in bulk and pharmaceutical dosage forms.

A Novel stability-indicating method was developed and validated by Reverse phase Hplc method   for the estimation of Thiotepa in Lyophilized form. The simple, Accurate Assay method was developed by PDA Detector at 215nm and column(YMC,150mmx4.6mm, 3.5 μm,12nm).The mobile phase A consisted of (3.5g of potassium Dihydrogen phosphate/1liter: 4.2g of Disodium hydrogen Phosphate/2liters)Buffer : Acetonitrile (85:15v/v) and The mobile phase B consisted of (3.5g of potassium Dihydrogen phosphate/1liter: 4.2g of Disodium hydrogen Phosphate/2liters)Buffer : Acetonitrile (50:50v/v) gradient flow rate at 1.0 mL/min for 30minutes. Samples was stored at 5°c temperature in sample cooler and Column oven temperature was maintained  in the method at 30°c  Respectively. The retention time of Thiotepa was 11.313 minutes. The proposed method was developed and validated with respect to specificity, accuracy, precision, linearity, Analyte Stability. Specificity of the Assay method shows no interference from Diluent and degrading products formed by alkaline, acidic, oxidative, Water Hydrolysis conditions. At Each level, %Recovery of Thiotepa in formulations was obtained to be in a range of 98-102%. The linearity of the assay was established in the range of 159.915-479.744µg/mL with correlation coefficient (R2) > 0.9999. This method was shows simple, precise, more accurate, stability indicating and reliable determination of Thiotepa for drug stability assay in pharmaceutical studies.

A simple, accurate, rapid and precise isocratic High performance liquid chromatographic (HPLC) method was developed and validated for the quantification of betaine in Achyranthes aspera. The method employs Reversed Phase Phenomenex (US) C18 Column 250 x 4.6mm I.D, 5µ Particle Size system consisted of prominence LC-20AT gradient pumps, Prominence SIL- 20A Auto sampler and a Prominence SPD-M20A Diode Array Detector (SHIMADZU), Japan and flow rate of 1.0 ml/min with a load of 20μl. Water and acetonitrile was used as mobile phase in the composition of 10:90. The detection was carried out at 242 nm. Linearity range was 0.32-0.48 mg/ml. Retention time of betaine was found to 8.42 min. The recovery studies 98-103%. This method was validated for accuracy, precision, linearity and Robustness as per ICH guidelines. The method was found to be specific, selective, and robust.