Dapaglifozin is a Sodium-glucose co-transporter-2 inhibitors work by inhibiting SGLT2, to prevent re absorption of glucose and facilitate its excretion in urine. Impurities in pharmaceuticals which are unwanted chemicals that remains with the Active Pharmaceutical Ingredients (APIs), or develop during stability testing, or develop during formulation or upon aging of both API and formulation. The presence of these unwanted chemicals even in small amounts may influence the efficacy and safety of the pharmaceutical products. It is necessary to estimate related impurities present in this drug. A simple and very sensitive method developed for estimation of impurities present in Dapagifozin formulation by Reverse Phase High Performance Liquid Chromatographic method. Method is capable to detect impurities in very low level (0.1µm/mL). Chromatographic separation of six different impurities was achieved on Agilent Zorbax Bonus RP-18 (250 x 4.6) mm, 5µm column using gradient elution method at 30°C column temperature and the detection was carried at 230 nm at a flow rate of 1.2mL/min. The method was validated as per ICH Q2 (R1) guideline along with stress studies.

Background: A simple, Accurate, precise method was developed for the simultaneous estimation of the Diphenhydramine and Bromhexine in tablet dosage form by RP-HPLC method.

Methods: Chromatogram was run through standard discovery 150 x 4.6 mm, 5m. Mobile phase containing Buffer 0.01N Potassium Dihydrogen Phosphate: Acetonitrile taken in the ratio 50:50 was pumped through column at a flow rate of 1 ml/min. Buffer used in this method was 0.01N Potassium Dihydrogen Phosphate and pH adjusted to 3.0 with dilute Orthophosphoric acid solution. Temperature was maintained at 30°C. Optimized wavelength selected was 225 nm. Retention time of Diphenhydramine and Bromhexine were found to be 2.458 min and 2.972.

Results: % Relative Standard Deviation of the Diphenhydramine and Bromhexine were found to be 0.5 and 0.3 respectively. % Recovery was obtained as 99.20% and 99.40% for Diphenhydramine and Bromhexine respectively. Limit of Detection, Limit of Quantitation values obtained from regression equations of Diphenhydramine and Bromhexine were 0.07, 0.20 and 0.11, 0.33 respectively. Regression equation of Diphenhydramine is y = 9539.x + 42940, and y = 9765x + 8034 of Bromhexine.

Conclusion: Since the retention time decreased the run time also decreased. So the method developed was simple and economical that can be applied successfully for simultaneous estimation of both Diphenhydramine and Bromhexine in bulk and combined tablet formulation.

Purpose: Although Methotrexate (MTX) is a commonly used therapeutic agent in the treatment of cancer, its use in high doses leads to some toxic effects. Thus, we have aimed that to develop and validate sensitive, fast, inexpensive High Performance Liquid Chromatography-UV method for monitoring MTX concentration in plasma samples which is applicable for routine clinical analysis.

Methods: Plasma was deproteinized with acetone and the chromatographic separation was performed on C18 column (250 x 4.6 mmx 5μm) using mobile phases composed of 0.05 M sodium phosphate buffer/ tetrahydrofuran (95:5) (pH=4.85) (mobile phase A) and 0.05 M sodium phosphate buffer/tetrahydrofuran (75:25) (pH=4.0) (mobile phase B) at a flow rate of 0.6 mL/min. Ultraviolet detection was done at 313 nm and at ambient temperature.

Results: Retention time for MTX was 7.78 minutes. The linearity is evaluated by a calibration curve in the concentration range of 1.0-50.0 μmol/L and presented a correlation coefficient of 0.9999. Precision of method within a day was 0.67-4.02 % and between days was 1.16-5.19%. The limits of detection and quantification achieved 0.1 and 0.9 μmol/L, respectively.

Conclusion: The fast and precise method enables to analyze large number of samples by using less mobile phase that makes it to be cost-effective. Also, this method is suitable for quantitation of MTX after infusion of high doses of this drug and has good accuracy, precision and quantitation limit.