P.V

A new selective, sensitive and Rapid Reverse phase-UPLC method was developed and validated to determine the known potential impurities present in Telmisartan (TL) and Hydrochlorothiazide (HC) in fixed dose combination drug product. The quantification was carried out by using Acquity UPLC, HSS T3 (100 × 2.1) mm, 1.8m column, with a flow rate of  0.5mL/min at 225 nm .The mobile phase consists of 0.1% ortho phosphoric acid pH adjusted to 2.6 with diluted sodium hydroxide as Mobile phase A and acetonitrile as Mobile phase B. Separation of the impurities was achieved within 10.0 minutes of run time. Typical retention times of TL and HC were found to be about 5.4 and 2.0 minutes respectively. The product was subjected to various degradation conditions and validated in terms of linearity, precision, accuracy, LOD, LOQ and robustness in accordance with ICH guidelines. The known impurities quantified in this study were HC imp-1 to 4 for Hydrochlorothiazide and TL imp-1 to 6 for Telmisartan. Recovery was established for all the impurities with respective to LOQ to 150%. The data supports that the newly developed method is capable to determine all the potential impurities of TL and HC.

A new rapid, sensitive and precise stability indicating HPLC method was developed on kromasil RP-C18 column (250mm X 4.6mm, 5µm), for the simultaneous analysis of Escitalopram and Clonazepam. Chromatographic separation was achieved at 229nm by using a mobile phase composed of methanol, acetonitrile and phosphate buffer (70: 28: 2, v/v). Retention time of escitalopram and clonazepam were found to be 2.67min and 3.46min respectively. Method validation was done as per the ICH guidelines. Good linearity was established in the concentration range of 50-300µg/ml and 2.5-15µg/ml by escitalopram and clonazepam respectively. Resolution of escitalopram and clonazepam was found to be 5.25. Percentage RSD of precision was found to be less than 2. The percentage assay of escitalopram and clonazepam in commercial formulation was found to be 99.18 and 99.22 respectively. Stability of the molecule was measured by inducing different stress conditions like acidic, basic, peroxide, thermal, light and UV light.

A Molecular docking study on novel Acetamide derivatives as specific Mono amino oxidase (MAO) A inhibitory agents was performed with a set of 40 compounds to analyze their inhibitory action.  For this, compounds were designed on the basis of available literature and used as Ligands for molecular interaction. The structure of molecular target Mono Amino Oxidase A (MAO A) was retrieved from the PDB database (PDB ID 2Z5X). For comparative analysis Clorgyline, a well-known specific MAO A inhibitor was taken as the standard. Computational docking analysis was performed using PyRx, AutoDock Vina option based on scoring functions. Among 40 compounds the top 11 hits were recognized as promising MAO A inhibitors, according to their docking scores and selected for further study of interaction and visualization. Phenyl sulphanyl derivative with chlorobenzyl amino moiety (Code AD31) showed an optimum binding affinity and stable complex with a molecular target MAO A  with the binding energy of -8.3  kcal/mol as compared to the standard (-7.6 kcal/mol). These results indicated that proposed modification in Acetamide derivatives may produce potent and specific MAO A inhibitors to treat depression with lesser side effects.

A simple, accurate and stability indicating high performance liquid chromatographic (HPLC) method was developed for the simultaneous estimation of Torsemide and Spironolactone in combined dosage form. Isocratic RP-HPLC separation was achieved on Kromasil RP- C18 column (250mm×4.6mm; 5µm) using methanol: acetonitrile: water in the ratio of 50:30:20 (v/v), pH6.8, at flow rate of1.0ml/min at ambient temperature. Quantization was achieved by UV detection at 235nm over the concentration range of 10-60μg/ml for torsemide and 25-150μg/ml for spironolactone with percentage recoveries of range 99.688-101.792 and 98.282-101.811for torsemide and spironolactone respectively. Different stress degradation studies like acidic, alkaline, peroxide, thermal etc were measured for both standard drugs and results found that the stress degradation conditions doesn’t affect the elution of the both the drugs and hence the developed method was found to be stability indicating method.

A novel, convenient, accurate, precise and reproducible reverse phase high performance liquid chromatography was developed and validated for the estimation of Vilazodone in bulk and pharmaceutical tablet dosage form. Objective was achieved under optimized chromatographic conditions on Shimadzu LC-20AT Prominence Liquid Chromatograph with Welchrom C18 isocratic column, (250 mm × 4.6 mm i.d., particle size 5 μm, maintained at ambient temperature), is used as stationary phase. An isocratic mode with mobile phase consisting of Acetonitrile: Water (50:50 v/v), with apparent pH of 3.3, at a flow rate of 1.0 mL/minutes. The effluent was monitored at 240 nm using Shimadzu SPD-20A prominence UV-Vis detector. The retention time of Vilazodone was found to be 4.103 minutes. The linearity range was found to be 1-5 μg/mL with correlation coefficient (R2) is 0.999. Validation parameters such as specificity, linearity, precision, accuracy, and robustness, limit of detection (LOD) and limit of quantitation (LOQ) were evaluated for the method according to the International Conference on Harmonization ICH Q2 (R1) guidelines. The LOD and the LOQ were found to be 0.044 μg/mL and 0.135 μg/mL respectively. Recovery of Vilazodone was found to be in the range of 99.80 % - 99.92 %. The method was validated statistically using the % RSD and the values are found to be within the limits. Therefore this method was conveniently and easily applied for the quantitative determination of Vilazodone in pharmaceutical dosage forms.

An accurate, simple and precise RP-HPLC method for the simultaneous estimation sulfadiazine and trimethoprim in pharmaceutical formulations was developed and validated. Chromatographic separation of two drugs was achieved on PEAK 7000 isocratic HPLC with rheodyne manual sample injector by using the mobile phase consisting of methanol, water and aceticacid in the ratio 70:25:05 (v/v/v) at a flow rate of 1mL/min and the wavelength of detection was at 237 nm. The retention time for sulfadiazine and trimethoprim were found to be 4.24 and 7.25 min respectively. The linearity of the method was tested over a concentration range of 41-287 µg/mL for sulfadiazine and 9-63 µg/mL for trimethoprim and the correlation coefficient was 0.999 for sulfadiazine and 0.998 for trimethoprim which is almost equal to 1. The limit of quantification was 3.5 μg/mL for sulfadiazine and 1 μg/mL for trimethoprim and the limit of detection was 1 μg/mL for sulfadiazine and 0.3 μg/mL for trimethoprim. The percentage recoveries were ranged from 98.64-101.64 for sulfadiazine and 98.56-100.86 for trimethoprim.

The concept of "Quality by Design" (QbD) is getting popularized in pharmaceutical manufacturing industry to understand the product and process to identify the risks involved during manufacturing.  One of the perpetual quality attribute is to have robust analytical method that can provide consistent results though out the life cycle of the product. General considerations during analytical method validation is to perform robustness studies by deliberate changes made in pH of the buffer in mobile phase, change in organic ratio, change in column oven temperature, change in buffer strength and using different column lot numbers etc. However to improve the analytical quality standard, a novel method concept under QbD was introduced which uses single mobile phase for three drug components and estimates using different column chemistries used in pharmaceutical industry viz., C18, C8, phenyl and Cyano column. The validated RP-LC method was successfully applied to the quantitative determination of Chlorpheniramine, Ibuprofen and Pseudoephedrine in tablet dosage form, helping to improve quality control and to assure therapeutic efficacy using all column chemistries.

To evaluate the anti-convulsion activity of Crocus sativus linn. aqueous extract  in electrical induced epilepsy models. To compare the anti convulsant activity of crocus sativus linn. with phenytion .After obtaining Institutional Ethical Committee approval, Wistar albino rats (150-200g) of either sex were randomly divided into 5 groups of 6 animals each. Dried powder of crocus sativus linn. was boiled with distilled water, cooled, filtered, placed on hotplate for complete evaporation, finally weighed and stored. The control group, test group and standard drugs group received saline, crocus sativus linn. extract (200,400 & 800 mg/kg), phenytoin (25 mg/kg) respectively by oral feeding. The anti-convulsent effect was assessed by maximal electrical shock (MES) in rats. In electrical induced epilepsy models It implies that saffron 400mg/kg (groupIV) and 800mg/kg(groupV) significantly (p

A new gradient reverse phase Ultra Performance Liquid Chromatography (UPLC) method was developed for the analysis of Dissolution profile samples of Paricalcitol in Paricalcitol Soft Gelatin capsules. The aim of the new method was to achieve proper accuracy and precision for the highly potent low dose drug product formulations. The normal injection loop allows upto 10µL of the sample in normal condition in UPLC systems.  For the current method the loop was modified to handle 50µL of injection volume in order to achieve quantifiable area counts. Efficient separation is achieved on Acquity UPLC HSS T3 (100 mm length × 2.1 mm ID with, 1.8 m particle size. Validation parameters such as specificity, linearity, precision, accuracy, and robustness were evaluated as per ICH guidelines. The validated RP-UPLC method was successfully applied to the Dissolution of Paricalcitol Soft Gelatin Capsules dosage forms.

This paper describes a strategy for the systematic development and validation of stability-indicating method of the determination of degradation impurities present in Naftopidil Orally dispersible tablets.  Efficient separation is achieved in 75mm length x 2.1mm ID, Octadecyl column with 3µ particle size. Using pH 3.2 phosphate buffer and acetonitrile as mobile phase in gradient pump mode. Flow rate was selected 0.4mL.min-1with a detection wavelength of 210nm. Validation parameters such as specificity, linearity, precision, accuracy, determination of LOD,  LOQ and robustness were evaluated as per ICH guidelines. The validated Reverse phase –Ultra Performance liquid chromatography (RP-UPLC) method was successfully applied to the quantitative determination of impurities of Naftopidil in Naftopidil Orally Dispersible tablet dosage forms, helping to improve quality control and to assure therapeutic efficacy at reduced run time of minutes.