Objective: The present study deals with the development, validation, and application of simple, precise, and accurate high-performance liquid chromatography (HPLC) method for the simultaneous estimation of metformin hydrochloride and ertugliflozin in pharmaceutical formulation and to validate.Methods: The analytical conditions were optimized on BDS C8 column (150 mm × 4.6 mm, 5 μm) at room temperature. The mobile phase consists of buffer: acetonitrile in 55:45 v/v ratio. Injection volume was 10 μl. The flow rate was maintained at 1.0 ml/min, and the analysis was carried out at 224 nm.Results: The method was found to be linear in the concentration range of 125–750 μg/ml and 1.875–11.25 μg/ml for metformin hydrochloride and ertugliflozin with regression coefficient r2 = 0.999. The method was found to be precise with percentage relative standard deviation below 2%. The limit of detection and limit of quantification were found to be within the limits. The percentage recovery of the developed method was 100.15%. All the validation parameters such as robustness, recovery, and precision were found to be within the limits. Degradation parameters such as acid, base, thermal and peroxide, light, temperature, and humidity were performed and found that the drugs are stable in all the extreme conditions.Conclusions: A simple, accurate, precise, and less time-consuming reversed-phase HPLC method for the simultaneous estimation of metformin hydrochloride and ertugliflozin has been developed and validated in accordance with the ICH guidelines.

Objective: The present study deals with the development, validation, and application of simple, precise, and accurate high-performance liquid chromatography (HPLC) method for the simultaneous estimation of metformin hydrochloride and ertugliflozin in pharmaceutical formulation and to validate.Methods: The analytical conditions were optimized on BDS C8 column (150 mm × 4.6 mm, 5 μm) at room temperature. The mobile phase consists of buffer: acetonitrile in 55:45 v/v ratio. Injection volume was 10 μl. The flow rate was maintained at 1.0 ml/min, and the analysis was carried out at 224 nm.Results: The method was found to be linear in the concentration range of 125–750 μg/ml and 1.875–11.25 μg/ml for metformin hydrochloride and ertugliflozin with regression coefficient r2 = 0.999. The method was found to be precise with percentage relative standard deviation below 2%. The limit of detection and limit of quantification were found to be within the limits. The percentage recovery of the developed method was 100.15%. All the validation parameters such as robustness, recovery, and precision were found to be within the limits. Degradation parameters such as acid, base, thermal and peroxide, light, temperature, and humidity were performed and found that the drugs are stable in all the extreme conditions.Conclusions: A simple, accurate, precise, and less time-consuming reversed-phase HPLC method for the simultaneous estimation of metformin hydrochloride and ertugliflozin has been developed and validated in accordance with the ICH guidelines.

Background The development of sound bioanalytical liquid chromatography-mass spectroscopy (LC-MS) method(s) is of paramount importance during the process of drug discovery and development, eventually culminating in marketing approval. The use of oral antidiabetic agents has been increased significantly from past decades, and till now, no bioanalytical method is available for quantitation of metformin (MET) and ertugliflozin (ERT) in the biological matrix that can be applied in bioequivalence studies using LC-MS/MS. Objective To study the use of highly responsive simple liquid–liquid extraction method development using deuterated MET and deuterated ERT, LC-MS/MS method for gradation of MET and ERT in the rat plasma. Materials and methods The chromatographic condition involves isocratic mode using Waters XBridge C18 3.5 μ (150×4.6 mm) column. Mobile phase was 0.1% orthophosphoric acid and acetonitrile in the ratio of 80 : 20 v/v. Detection was carried out on a triple quadrapole MS employing electrospray ionization technique, operating multiple reactions, monitoring with the transitions of m/z 258.2→174.1, m/z 250.1→210.2, m/z 258.2→174.1, and m/z 260.3→210.2 for MET, ERT, deuterated MET, and deuterated ERT, respectively, in the positive ion mode. Results and conclusion The method has been validated, and the linearity was observed in the range of 10–150 ng/ml and 0.1–1.5 ng/ml for MET and ERT, respectively. For intraday and interday %RSD, the values were found to be within the acceptable limits. Recovery studies for MET and ERT obtained, mean recovery of 99.5 and 98.6%, respectively. A battery of stability studies like bench-top stability, autosampler stability, freeze-thaw stability, and long-term stability were performed. Highly responsive simple LC-tandem MS assay method was developed and witnessed for the gradation of MET and ERT in the rat plasma; the developed method was applied to pharmacokinetic studies.

A fascinating oncology target is the serine/threonine-specific protein kinase B-Raf, which is a component of the MAPK pathway. A new series of triazolo oxadiazole derivatives have been identified as the primary scaffold of small molecule B-Raf inhibitors. Twelve derivatives (7a-l) of triazolo oxadiazole hybrids were synthesized and characterized by spectral data. The evaluation of physicochemical and pharmacokinetic parameters demonstrated that the compounds' solubility, lipophilicity, and compliance with Lipinski's rule are all within acceptable ranges. The anti-proliferative capability of the synthesized derivatives was evaluated by screening them against the human melanoma cell lines Skmel-23 and A375, which express B-Raf in their wild-type and mutant forms, respectively. The IC 50 values differed significantly from those of the conventional doxorubicin. The IC 50 values for compounds 7b and 7e against Skmel-23 were determined to be 26.95 and 76.17 μM, respectively. The compounds 7b and 7c inhibited A375 cells at IC 50 concentrations of 16.64 and 78.1 μM, respectively. Therefore, these compounds were subjected to morphological screening to detect changes in cell morphology and evaluate their apoptotic potential. The compound 7b was found to significantly inhibit both cell lines and is therefore a promising therapeutic candidate for targeting both wild-type and mutant B-Raf forms.

B‐Raf, a proto‐oncogene that encodes the B‐Raf protein in the Ras/Raf/Mek/Erk (MAPK) pathway, is important in directing cell development and differentiation, mutation of which results in various types of cancers. A series of novel indazole derivatives were synthesized employing scaffold‐based approach and characterized by the spectral data. All the synthesized compounds were subjected to molecular docking to assess their binding affinity with the protein. The physicochemical and pharmacokinetic profile of the compounds revealed their compliance with Lipinski's rule. The antiproliferative capacity of the synthesized derivatives was assessed by screening them against the human melanoma cell lines Skmel‐23 and A375, which express B‐Raf in wild‐type and mutant forms, respectively. Compounds with 2‐chloro and 2‐cyano‐4‐isoproxy substitutions elicited potent inhibition against melanoma cell lines with IC50 values of 40.11 and 13.37 μM against Skmel‐23 and 16.89, 15.57 μM against A375 cells respectively. The compounds manifested greater potential than the standard doxorubicin. In order to detect changes in cell morphology and evaluate their apoptotic potential; these substances were subjected to morphological screening. Both cell lines were shown to be strongly inhibited by the 2‐cyano‐4‐isoproxy substituted indazole derivative; making it a prospective therapeutic candidate for targeting both wild‐type and mutant B‐Raf forms.