Development of Robust RP-HPLC Method for Concurrent Analysis of Aliskiren and Amlodipine in Combined Tablet Dosage Form

Development and validation of RP-HPLC method for simultaneous estimation of Atazanavir and Ritonavir in their combined tablet dosage form. Atazanavir and Ritonavir are antiviral agents used in treatment of HIV. A simple, precise, rapid, accurate and cost-effective high- performance liquid chromatography (HPLC) method was successfully developed and validated for simultaneous estimation of Atazanavir and Ritonavir in their combined tablet dosage form. The selected mobile phase was Methanol: Phosphate Buffer in proportion 65:35 v/v respectively. The optimized columns used are C18 column, Symmetry and Zodiac column. X bridge C18 (4.6×150 mm, 5 µm) particle size was found to be ideal as it gave good peak shape and resolution at 1ml/min flow for Atazanavir and Ritonavir. In this study, the validation of Atazanavir and Ritonavir in API and marketed formulations were performed keeping in accordance with the parameters like system suitability, specificity, linearity, accuracy, precision (reproducibility & repeatability), robustness. The developed stability indicating method is capable for determination of impurities of Atazanavir and Ritonavir in combined tablet dosage form as well as individual dosage forms also. The method has been successfully validated according to ICH guidelines and the results obtained by using RP – HPLC are rapid, accurate and precise. The proposed methods were successfully applied for the analysis of synthetic mixtures and pharmaceutical formulations of Atazanavir and Ritonavir.

A simple, robust, precise and accurate reverse phase liquid chromatographic method was developed for simultaneous estimation of Atazanavir sulphate and Ritonavir in combined tablet dosage form by RP-HPLC method. Chromatography was carried out on a Nucleodur C18Â column (150 mm x 4.6 mm x 5 m) using Acetonitrile: Methanol: Phosphate buffer, pH was adjusted to 3.0 with orthophosphoric acid in the ratio of 44:11:45 (V/V) as a mobile phase at a flow rate of 1.5 mL min-1Â and eluents were monitored at 210 nm. The calibration curves were linear over the range of 34 102 ?g mL-1Â for Atazanavir sulphate and 10 30 ?g mL-1Â for Ritonavir. The average retention time of Atazanavir sulphate and Ritonavir was found to be 3.133 min and 6.133 min respectively. The method was reproducible, with good resolution of Atazanavir sulphate and Ritonavir. The results of the analysis have been validated statistically.

Development and validation of eco-friendly micellar-HPLC and HPTLC-densitometry methods for the simultaneous determination of paritaprevir, ritonavir and ombitasvir in pharmaceutical dosage forms

The present paper describes a reverse phase HPLC method for simultaneous estimation of Trifluoperazine and Isopropamide from their combined tablet dosage form. The proposed RP-HPLC method utilizes an Agilent Zorbax, C18 column (250 × 4.6 mm, 5μm particle size) using a mobile phase consisting of mixture of Buffer: Acetonitrile (80: 20 v/v) mobile phase flow rate of 0.8 ml/min; and UV detection at 227nm. The retention time for Trifluoperazine and Isopropamide were 2.4 and 3.62 minutes respectively. The method was linear in the range of 40-120μg/ml and 100-300μg/ml and correlation coefficients were 1.00 and 1.00 for Trifluoperazine and Isopropamide respectively. The limit of detection was found to be 2.963 and 2.985µg/ml for Trifluoperazine and Isopropamide respectively. Limit of quantification was found to be 9.877 and 9.9502 for Trifluoperazine and Isopropamide respectively. The percentage recoveries for Trifluoperazine and Isopropamide ranged from 100.03-100.36% and 100.00-100.07 % respectively. The percentage of RSD for precision of the method was found to be less than 2%. The proposed method can be used for the routine analysis for the estimation of these drugs in combined dosage form.

A simple and selective LC method is described for the determination of Atorvastatin, Ezetimibe and Fenofibrate in tablet dosage forms. Chromatographic separation was achieved on a reversed-phase C18 column (Inertsil ODS 3 V, 5 μ, 250 mm × 4.6 mm) using mobile phase of a mixture of 80 volumes of Methanol: 10 volumes of Acetonitrile and 10 volumes of Water with detection of 256 nm. Linearity was observed in the range 3-7 μg/ml for Atorvastatin (r2=0.9985), 3-7 μg /ml for Ezetimibe (r2=0.9971) and 48-112 μg /ml for Fenofibrate (r2=0.9964) for the amount of drugs estimated by the proposed methods was in good agreement with the label claim. The results show that the method was found to be specific, simple, accurate, precise, sensitive and validated. The accuracy of the methods was assessed by recovery studies at three different levels. The method was found to be precise as indicated by the repeatability analysis, showing %RSD less than 2. The method was successfully applied for the determination of three drugs in combined tablet dosage form.

The aim of this work is to develop and validate a simple, sensitive, rapid and accurate and less time consuming validated RP-HPLC method for Etodolac and Thiocolchicoside in pharmaceutical dosage form. The method was developed and validated for various parameters as per ICH guidelines. The results obtained were within the acceptance criteria. The proposed method was applied for the determination of Etodolac and Thiocolchicoside in marketed formulation. The assay results confirm with the label claim of formulation.. Hence, the proposed method was found to be satisfactory and could be used for the routine analysis of Etodolac and Thiocolchicoside in combined tablet dosage forms. The retention time were found to be was 2.777 and 2.380 min for Etodolac and Thiocolchicoside respectively. The quantitative estimation gave a satisfactory result for Etodolac (99.66 % w/w) and Thiocolchicoside (99.3% w/w) respectively. The regression values over its peak areas were found to y = 17537x + 361.5 and y = 17769x + 1505 respectively and correlation coefficient found to be about 0.9999 and 0.9998 for Etodolac and Thiocolchicoside respectively. The percentage recovery for Etodolac and Thiocolchicoside were found to be 99.99% and 101.2 % respectively

BackgroundAn accurate, precise and robust analytical method was developed for the impurity profiling in the metformin hydrochloride and teneligliptin hydrobromide hydrate tablet. The gradient was optimized for better separation of impurities by using BDS Hypersil C18 250 × 4.6 mm, 5µ column operated at 35 °C. The octane sulfonic acid and phosphate buffer with triethylamine at pH 3.0 were used as mobile phase A, and acetonitrile was used as mobile phase B. The mobile phase was pumped at 1.0 mL/min. The gradient was optimized for better resolution, and the chromatogram was monitored at 210 nm.ResultsThe % recovery of teneligliptin and metformin HCL observed was above 90% from LOQ level to 150%. The correlation coefficient r2 was 0.999 for metformin HCl, teneligliptin, melamine, cyanocobalamin, teneligliptin impurity A and 0.998 for teneligliptin impurity B. The method was found unaffected by change in method variance during the robustness study. During the stress study with acid, base, peroxide and temperature, maximum degradation was observed with peroxide indicating the sensitivity of the molecule toward oxidative stress.ConclusionsThe developed method is precise, accurate, robust and linear and hence can be routinely used for the related substance analysis of metformin hydrochloride and teneligliptin hydrobromide hydrate tablet in the quality control laboratory at manufacturing site during the commercial manufacturing.

The study describes method development and subsequent validation of RP-HPLC method for simultaneous estimation of emtricitabine, tenofovir disoproxil fumarate and rilpivirine in combined tablet dosage forms. Chromatographic separation was achieved on a hypersilBDSC18 column (250 mm x 4.6 mm, 5 µm) using a mobile phase consisting of (45:55 v/v) buffer: acetonitrile at a flow rate of 1 mL/min. The detection wavelength is 280 nm. The retention times of emtricitabine, tenofovirdisoproxil fumarate and rilpivirine were found to be 2.692, 4.402 min and 5.725 min respectively. The developed method was validated as per ICH guidelines. The developed and validated method was successfully used for the quantitative analysis of emtricitabine,tenofovirdisoproxilfumarate and rilpivirine in tablet dosage forms.

A rapid and precise reverse phase high performance liquid chromatographic method has been developed for the validated of Trihexyphenidyl and Haloperidol, in its pure form as well as in tablet dosage form. Chromatography was carried out on a Altima C18 (4.6 x 150mm, 5μm) column using a mixture of Methanol: TEA Buffer pH 4.5: Acetonitrile (50:25:25) as the mobile phase at a flow rate of 1.0ml/min, the detection was carried out at 225 nm. The retention time of the Trihexyphenidyl and Haloperidol was 2.102, 3.537±0.02min respectively. The method produce linear responses in the concentration range of 15-75ppm of Trihexyphenidyland 37.5-187.5ppm of Haloperidol. The method precision for the determination of assay was below 2.0%RSD. The method is useful in the quality control of pharmaceutical formulations.

Development and validation of RP-HPLC method for determination of Atorvastatin calcium and Nicotinic acid in combined tablet dosage form

A simple, sensitive and accurate RP-HPLC method for the simultaneous quantification of ambroxol, roxithromycin and serratiopeptidase in bulk and combined tablet dosage form was developed and validated. Ambroxol, roxithromycin and serratiopeptidase were separated and estimated using Waters HPLC system and YMC Pack pro C18 (250 × 4.6 mm, 5 \(\mu\)m particle size) column. The mobile phase consisted of 0.1% orthophosphoric acid and acetonitrile (60:40 v/v). The calibration curves were linear over concentration range of 12-36 \(\mu\)g/mL, 60-180 \(\mu\)g/mL and 6-18 \(\mu\)g/mL with limits of detection of 0.040 \(\mu\)g/mL, 1.176 \(\mu\)g/mL and 0.127 \(\mu\)g/mL for ambroxol, roxithromycin and serratiopeptidase, respectively. For the studied drugs, recovery varied in range of 99.57% - 100.24% with relative standard deviation ranging from 0.12% to 0.36%. The developed method was validated as per the ICH guidelines. The proposed RP-HPLC method could be used for the analysis of ambroxol, roxithromycin and serratiopeptidase simultaneously in combined tablet dosage forms.

A RP-HPLC method for the estimation of ATN (Atenolol) and CTN (chlorthalidone) in combined dosage form was developed using Comosil RP-C18 (4.6 x 250mm, 5m) in an gradient mode with mobile phase comprising of Methanol: Water (pH 3 using OPA) The flow rate was 1 mL/ min and effluent was monitored at 226.0 nm. The retention times were found to be 2.2 min for ATN and 3.36 min for CTN. The assay exhibited a linear dynamic range of 40- 200 g/mL for ATN and 10- 50 g/mL for CTN. The calibration curves were linear (r 2 = 0.999 for ATN and r 2 = 0.999 for CTN) over the entire linear range. Mean % recovery was found to be 99.78 % for ATN and 99.30 % for CTN with % RSD was NMT 2 for both estimations which fully agrees with system suitability which is in good agreement with labeled amount of formulation. The % RSD for Intra- Day & Inter-Day Precision was NMT than 2 for both the drugs. The developed method was validated as per ICH guidelines

This work intends to develop a QbD-based RP-HPLC method and validate rutin (RU) and ascorbic acid (ASC) in combined tablet dosage form. The method is optimized by the central composite design. The independent variables chosen are mobile phase ratio and pH. The dependent variables are retention time for RU (R1), retention time for ascorbic acid ASC (R2), and resolution (R3). Analysis of variance revealed that the method parameters were significant (p <5). Waters Alliance-e2695 [C18 (150x 4.6 mm, 3.5)] was used to separate rutin and ascorbic acid using a mobile phase Acetonitrile: Hexane Sulphonic Acid (pH-2.5/OPA) in a 70:30 ratio, the selected wavelength was 215 nm. Method validation and degradation studies were performed ICH guidelines are followed. The approach was determined to be easy, suitable, accurate, precise, and robust for quantitative analysis of ascorbic acid and rutin.

A force degradation profile of Metformin Hcl & Glimepiride in combine tablet dosage form on RP-HPLC was developed using Grace RP-C18 (4.6 x 150 mm, 5 m) in an gradient mode with mobile phase comprising of Acetonitrile: Dihydrogen Pott.Phosphate (pH 2.5 using 0.1% OPA) The flow rate was 0.7 mL/ min and effluent was monitored at 242 nm.. The retention times were found to be 2.06 min for MET and 5.80 min for GLIM. The assay shows a linear dynamic range of 250- 1250 g/mL for MET and 1.0-5.0 g/mL for GLIM. The calibration curves were linear (r 2 = 0.999 for MET and r 2 = 0.998 for GLIM) over the entire linear range. Mean % recovery was found to be 99.80 % for MET and 98.93 % for GLIM with % RSD was NMT 2 for both estimations which fully agrees with system suitability which is in good agreement with labeled amount of formulation. The % RSD for Intra- Day & Inter-Day Precision was NMT than 2 for both the drugs. The developed method was validated as per ICH guidelines

A high performance reverse phase liquid chromatographic procedure is developed for simultaneous estimation of metformin hydrochloride and pioglitazone hydrochloride in combined tablet dosage form. The mobile phase used was a combination of acetonitrile:water:acetic acid (60:40:0.3) and the pH was adjusted to 5.5 by adding triethylamine. The detection of the combined dosage form was carried out at 230 nm and a flow rate employed was 1 ml/min. Linearity was obtained in the concentration range of 0.015 to 0.120 μg/ml of pioglitazone hydrochloride and 0.5 to 4.0 μg/ml of metformin hydrochloride with a correlation coefficient of 0.9992 and 0.9975. The results of the analysis were validated statistically and recovery studies confirmed the accuracy and precision of the proposed method.