METHOD DEVELOPMENT AND VALIDATION OF ULTRAVIOLET-VISIBLE SPECTROSCOPIC METHOD FOR THE ESTIMATION OF HEPATITIS-C DRUGS - DACLATASVIR AND SOFOSBUVIR IN ACTIVE PHARMACEUTICAL INGREDIENT FORM

ABSTRACTObjective: The objective of the present work is to develop a simple, efficient, and reproducible spectrophotometric method for the quantitativeestimation of hepatitis-C drugs - Daclatasvir and Sofosbuvir in its active pharmaceutical ingredient (API) form.Methods: The developed ultraviolet spectrophotometric method for the quantitative estimation of hepatitis-C drugs - Daclatasvir and Sofosbuvir isbased on measurement of absorption at a wavelength maximum (λmax) of 317 and 261 nm using methanol as solvent.Results: The method was validated in terms of specificity, precision, linearity, accuracy, and robustness as per the ICH guidelines. The method wasfound to be linear in the range of 50-150% for Daclatasvir and in the range of 43-143% for Sofosbuvir. The percentage recovery values were in therange of 99.4-100.6% for Daclatasvir and in the range of 99.7-100.6% for Sofosbuvir at different concentration levels. Relative standard deviation forprecision and intermediate precision results were found to be <2%. The correlation coefficient value observed for Daclatasvir and Sofosbuvir drugsubstances was not <0.99, 0.99, respectively. Results obtained from the validation experiments prove that the developed method is quantified for theestimation of Daclatasvir and Sofosbuvir drug substances.Conclusion: The developed method can be successfully applied for routine analysis, quality control analysis, and also suitable for stability analysis ofDaclatasvir and Sofosbuvir in API form as per the regulatory requirements.Keywords: Daclatasvir, Sofosbuvir, Method development, Validation, Ultraviolet-visible spectrophotometry

STABILITY INDICATING REVERSE PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHIC METHOD FOR SIMULTANEOUS ESTIMATION OF LABETALOL AND ITS DEGRADATION PRODUCTS IN TABLET DOSAGE FORMS

ABSTRACTObjective: The objective of the present work is to develop a simple, efficient, and reproducible stability indicating reverse phase high-performanceliquid chromatographic method for simultaneous determination labetalol and its degradation products in tablet dosage forms.Methods: The chromatographic separation of labetalol and its degradation products in tablets was carried out on Zorbax Eclipse Plus C-18(100 × 4.6 mm, 3.5 µm) column using 0.1% trifluoroacetic acid (TFA) (v/v) in 1000 ml of water and 0.1% TFA (v/v) in 1000 ml of acetonitrile:Methanol (1:1) by linear gradient program. Flow rate was 1.0 mL min with a column temperature of 35°C, and detection wavelength was carried outat 230 nm. Known impurity is well resolved from the main active drug within 14 minutes run time.−1Results: The forced degradation studies were performed on labetalol tablets under acidic, basic, oxidation, thermal, humidity, and photolyticconditions. No degradation products were observed from the forced degradation studies, and the known impurity is well resolved from the mainactive drug. The method was validated in terms of specificity, linearity, limit of detection (LOD), limit of quantitation (LOQ), accuracy, precision, androbustness as per the ICH guidelines. The method was found to be linear in the range of LOQ to 120% for all the known and unknown impurities.The LOD and LOQ values of known impurity were found between 0.3593 and 0.7187 µg mL, and the percentage recovery values were in the rangeof 95.5-105.2% at different concentration levels. Relative standard deviation for precision and intermediate precision results were found to be <5%.The correlation coefficient found for all compounds was not <0.99. The results obtained from the validation experiments prove that the developedmethod is a stability indicating method.−1Conclusion: The developed method can be successfully applied for routine analysis, quality control analysis and also suitable for stability analysis ofthe simultaneous determination of labetalol and its degradation products in tablet dosage forms as per the regulatory requirements.Keywords: Labetalol, Development, Validation, Reverse phase high-performance liquid chromatography

DEVELOPMENT AND VALIDATION OF A DISSOLUTION METHOD FOR FROVATRIPTAN TABLETS BY REVERSED PHASE UPLC

Objective: The main objective of the method was to develop a simple, rapid, efficient and reproducible, stability indicating reverse phase ultra performance liquid chromatography (RP-UPLC) method for the estimation of frovatriptan in tablet dosage form.Methods: The RP-UPLC method for estimation of frovatriptan (FRT) in their tablets was carried out on Acquity UPLCTM, BEH C-18 (100 × 2.1 mm, 1.7 µm) column using 0.1% trifluroacetic acid buffer and a mixture of methanol and acetonitrile (50:50) using isocratic program. The flow rate of the mobile phase was 0.2 mL min-1and detection wavelength was carried out at 244 nm. Total runtime is 3 minutes for chromatographic run. The method was validated in terms of specificity, linearity, accuracy, precision and robustness as per ICH guidelines.Results: The method was found to be linear in the range of 1.41-3.67 μg mL-1. Recovery was found to be in the range of 97.8-101.8%. Relative standard deviation for precision and intermediate precision was found to be less than 3%. The developed method was successfully applied for the estimation of frovatriptan in tablet formulation and average dissolution rate was found to be 93%. The results obtained from the validation experiments prove that the developed method is suitable for routine analysis.Conclusion: The developed RP-UPLC method was simple, rapid, accurate, and precise for the estimation of dissolution rate in frovatriptan tablet dosage form.Â

METHOD DEVELOPMENT AND VALIDATION OF UV-VISIBLE SPECTROSCOPIC METHOD FOR THE ESTIMATION OF ASSAY OF SUGAMMADEX SODIUM, APREMILAST, RIOCIGUAT AND VORAPAXAR SULFATE DRUGS IN API FORM.

Objective: The objective of the present work is to develop a simple, efficient and reproducible spectrophotometric method for the quantitative estimation of Sugammadex Sodium, Apremilast, Riociguat and Vorapaxar sulfate drugs in its active pharmaceutical ingredient (API) form.Methods: The developed UV-Visible spectrophotometric method for the quantitative estimation of drugs – Sugammadex Sodium, Apremilast, Riociguat and Vorapaxar sulfate is based on measurement of absorption at a wavelength maximum (λmax) of 210nm, 230nm, 323nm and 271nm using water and methanol as diluents.Results: The method was validated in terms of specificity, precision, linearity, accuracy, and robustness as per the ICH guidelines. The method was found to be linear in the range of 33% to 167% for Sugammadex Sodium and Apremilast drug substances; 50% to 150% for Riociguat and Vorapaxar sulfate drug substances. The percentage recovery values were in the range of 99.7 to 100.9% for Sugammadex Sodium, 99.3 to 100.3% for Apremilast, 99.7 to 100.3% for Riociguat and in the range of 99.5 to 100.3% for Vorapaxar sulfate at different concentration levels. Relative standard deviation for precision and intermediate precision results were found to be less than 2%. The correlation co-efficient value observed for Sugammadex Sodium, Apremilast, Riociguat and Vorapaxar sulfate drug substances was not less than 0.99 for their respective drugs. Results obtained from the validation experiments prove that the developed method is quantified for the estimation of assay of Sugammadex Sodium, Apremilast, Riociguat and Vorapaxar sulfate drug substances.Conclusion: The developed method can be successfully applied for routine analysis, quality control analysis and also suitable for stability analysis of assay of Sugammadex Sodium, Apremilast, Riociguat and Vorapaxar sulfate in API form as per the regulatory requirements.Keywords: Sugammadex Sodium, Apremilast, Riociguat, Vorapaxar sulfate, Method Development, Validation, UV-Visible spectrophotometry

Stability-Indicating RP-HPLC Method for Simultaneous Estimation of Enrofloxacin and Its Degradation Products in Tablet Dosage Forms

The present work was the development of a simple, efficient, and reproducible stability-indicating reverse-phase high performance liquid chromatographic (RP-HPLC) method for simultaneous determination enrofloxacin (EFX) and its degradation products including ethylenediamine impurity, desfluoro impurity, ciprofloxacin impurity, chloro impurity, fluoroquinolonic acid impurity, and decarboxylated impurity in tablet dosage forms. The separation of EFX and its degradation products in tablets was carried out on Kromasil C-18 (250×4.6 mm, 5 μm) column using 0.1% (v/v) TEA in 10 mM KH2PO4 (pH 2.5) buffer and methanol by linear gradient program. Flow rate was 1.0 mL min−1 with a column temperature of 35°C and detection wavelength was carried out at 278 nm and 254 nm. The forced degradation studies were performed on EFX tablets under acidic, basic, oxidation, thermal, humidity, and photolytic conditions. The degraded products were well resolved from the main active drug and also from known impurities within 65 minutes. The method was validated in terms of specificity, linearity, LOD, LOQ, accuracy, precision, and robustness as per ICH guidelines. The results obtained from the validation experiments prove that the developed method is a stability-indicating method and suitable for routine analysis