Stability-indicating RP-HPLC Method Research Articles

Stability-indicating RP-HPLC method development and validation for determination of nine impurities in apixaban tablet dosage forms. Robustness study by quality by design approach.

A quality by design (QbD) based high-resolution HPLC method is described for determination of impurities in apixaban (APX) in the tablet dosage form. Employing a simple and stability-indicating HPLC method, nine known impurities were quantified with good peak resolution. Mobile phase A (MP-A) was prepared with buffer and acetonitrile 90:10v/v, while mobile phase B (MP-B) contained water and acetonitrile 10:90 v/v. The gradient program was 0 min, MP-A 75%, B 25%; 20 min, MP-A 65%, B 35%; 30 min, MP-A 40%, B 60%; 40min, MP-A 40%, B 60%; 42 min, MP-A 75%, B 25%; and 50 min, MP-A 75%, B 25%. The chromatographic separation was achieved using a Zorbax RX C18 250 × 4.6 mm column, 5 μm (1.0 ml min-1 , 280 nm, 50 μl) and a column temperature of 40°C. Several separation studies were carried out using design of experiments to optimize the method. Validation results confirm the applicability of the developed method for quality analysis and stability studies of the regular product on the manufacturing stream.

Stability-Indicating RP-HPLC Method for the Quantification of Paliperidone in Bulk and Solid Dosage Form to Establish Validation and Stability Indicating Parameters

Stability-Indicating RP-HPLC Method for the Quantification of Paliperidone in Bulk and Solid Dosage Form to Establish Validation and Stability Indicating Parameters

Identification, Characterization and HPLC Quantification of Process-Related Impurities in Bepotastine Besilate Bulk Drug.

Bepotastine besilate (here after referred to as BTST), chemically known as ({d(S)4[4[(4chlorophenyl) (2pyridyl) methoxy] piperidino} butyric acid monobenzene sulphonate), is a second-generation antihistamine drug. To the best of our knowledge, no studies concerning the isolation or identification of process-related impurities have been reported so far. The current study reports the development and validation of a stability-indicating RP-HPLC method for the separation and identification of 5 potential impurities in bepotastine besilate. In this experiment, the structures of 3 process-related impurities were found to be new compounds. They were characterized and confirmed by NMR and MS spectroscopy analyses. These 3 new compounds were proposed to be (S)-4-[(phenyl)-2-pyridinylmethoxy]-1-piperidinebutanoic acid,(Imp-A); 4-[(S)-(4-chlorophenyl)-2-pyridinylmethoxy]-1- piperidinebutyric acid, N-oxide (Imp-B) and (S)-4-[(4- chlorophenyl)-2-pyridinylmethoxy]-1-piperidylethane (Imp-C). In addition, an efficient optimized chromatographic method was performed on a Shimadzu Inertsil C8-3 column (150 mm×4.6 mm, 3 μm) to separate and quantify these 5 impurities. It was using 15 mmol ammonium formate buffer in water (pH adjusted to 3.8 with formic acid) and acetonitrile as the mobile phase in gradient mode. The method was developed to separate and quantify these 5 impurities obtained in the range of 0.05-0.75 μg/mL. It was validated and proven to be selective, accurate and precise and suitable. It is the first publication of identification and characterization data of the 3 new compounds. It is also the first effective HPLC method for separation and quantification of all of process-related impurities in bepotastine besilate.

Development and Validation of a Stability-Indicating Ion-Pair RP-HPLC Method for Determination of Bethanechol with UV Detection: Application to Pharmaceutical Analysis

A rapid and stability indicating ion-pair RP-HPLC method was developed for estimation of bethanechol chloride (BC) in pharmaceutical preparations. The separation was achieved on a phenyl column using a mobile phase consisting of 0.05 M phosphate buffer (pH 6) – ethanol (98:2 v/v) mixture containing 0.56 mg/mL of sodium 1-heptanesulfonate. Quantitation was achieved with UV detection at 190 nm. In forced degradation studies, the drug was subjected to oxidation, hydrolysis, photolysis, and heat. The method was validated for specificity, selectivity, linearity, precision, accuracy, and robustness. The analytical procedure was found to be linear in a BC concentration range of 5.0 – 500.0 μg/mL (r2 = 0.9997). Precision was evaluated by replicate analysis in which%deviations from relative standard values of peak areas were found below 2.0. The recoveries obtained (99.0 – 102.0%) ensure the accuracy of the proposed method. Impurities as well as excipients were well resolved from the parent drug. Accordingly, the proposed validated procedure was proved to be suitable for routine and stability analysis of BC in pharmaceutical preparations.

Development and validation of a stability-indicating RP-HPLC method of cholecalciferol in bulk and pharmaceutical formulations: Analytical quality by design approach

Development and validation of a stability-indicating RP-HPLC method of cholecalciferol in bulk and pharmaceutical formulations: Analytical quality by design approach

HPLC METHOD DEVELOPMENT, VALIDATION, AND FORCED DEGRADATION FOR SIMULTANEOUS ANALYSIS OF OXYCODONE AND NALTREXONEIN PHARMACEUTICAL DOSAGE FORM

A simple, precise and stability-indicating RP-HPLC method was developed for simultaneous quantification of oxycodone (OXCD) and naltrexone (NTRX) in combined dosage form. The developed method was validated with respect to precision, linearity, accuracy, robustness, ruggedness, sensitivity and solution stability. The method was developed with ammonium di hydrogen phosphate buffer (pH 5.0) and acetonitrile in a ratio of 55:45 (v/v) as mobile phase at a flow rate of 1.0 mL/minute over Intersil ODS C18 column (250 mm × 4.6 mm×5μ).The UV detection wavelength was fixed at 235 nm. The column temperature was maintained at ambient temperature. The method showed good linearity with correlation coefficient values of 0.9990 and 0.9994 for OXCD and NTRX. The percent recoveries of the two drugs found within the limits of 98.0–102.0%. The LOQ concentrations of OXCD and NTRX are 0.625 μg/ mL and 0.075 μg/mL, respectively. The LOD concentrations of OXCD and NTRX are 0.3125 μg/mL and 0.0375 μg/mL, respectively. According to ICH guidelines forced degradation study was validated.

Quantitative Determination of Brexpiprazole by RP-HPLC Method

A stability-indicating RP-HPLC method was established and validated for the determination of Brexpiprazole in bulk drug using C18 column Waters (150 mm×4.6 mm, 5 µm) with a mobile phase containing of 500 mL of 10 mM monobasic Potassium phosphate buffer adjusted pH 2.0 with 85% Orthophosphoric acid and 500 mL of HPLC grade Acetonitrile. The mobile phase was filtered with a 0.45 µm membrane filter and degassed by sonicating for few minutes. The detection was performed at 213 nm using a Photodiode Array Detector at a flow rate of 1.0 mL min-1 and Brexpiprazole was eluted at 2.5 min. with column temperature 30°C. The detector response was generated from the concentration range of 0.01-0.06 mg mL-1 and regression coefficient(r) was 0.999. Brexpiprazole was exposed to stress conditions such as acidic, basic, oxidation, hydrolysis, photolysis and thermal degradation, and the outcomes showed that the molecule was more sensitive to peroxide degradation. This method was validated as per ICH and FDA guidelines and showed linearity, accuracy, precision, specificity, robustness, LOD, LOQ and system suitability results within the acceptance criteria.

Development and Validation of a Stability-indicating RP-HPLC Method for Estimation of Metformin and Rosuvastatin along with Impurities from a Combined Oral Solid Dosage Form

The objective of current study was to develop and validate a stability-indicating reversed-phase high-performance liquid chromatography for simultaneous quantification of impurities in metformin sustained release-rosuvastatin immediate release combined oral solid dosage form as per ICH guidelines. Chromatographic separation was achieved using a Waters HPLC system with a YMC-Pack ODS-A 100×4.6 mm; 3 µm column. Mobile phase A comprised of 20 mM ammonium acetate:acetonitrile (9:1 v/v) pH 3.5, and mobile phase B, consisted of 20 mM ammonium acetate:acetonitrile:methanol (3:5:2 v/v/v) pH 3.5. The gradient program was set as (T/% B) 0/30, 5/30, 10/50, 30/50, 40/80, 42/30 and 45/30, which pumped at a rate of 0.9 ml/min. About 10 μl of sample solutions were injected and monitored at 240 nm. Column temperature and sample compartment was maintained at 25° and 5°, respectively. Forced degradation studies revealed that the peak threshould was more than the peak purity and no purity flag was observed. Repeatability, intra, and inter-day precision results were well within the tolerable limits. Limits of detection were found to be 0.10, 0.05, 0.15, 0.20 and 0.40 ppm and limits of quantitation as 0.30, 0.15, 0.45, 0.60 and 1.0 ppm for metformin, rosuvastatin, rosuvastatin impurity A, B, and C, respectively. The correlation coefficient of linearity for metformin, rosuvastatin and its impurities were found to be ≥0.999. The method recovery was found between 99 and 102 %. The proposed method appeared to be a simple, rapid, economical, accurate, specific, robust and able to distinguish the two key components from their degradants, impurities, and excipients. Therefore, current method could be applied successfully for stability and routine examination.

Development and Validation of A Fast, Simple And Specific Stability Indicating RP-HPLC Method for Determination of Dexpanthenol in Eye Gel Formulation.

In this study a simple and reliable stability-indicating RP-HPLC method was developed and validated for analysis of Dexpanthenol in an artificial tear formulation. The chromatographic separation was performed on a HPLC C18 column (25.0 cm ´4.6 mm, 5 mm) using a mixture of 0.037 M monobasic potassium phosphate in water (adjusted with 0.1% (v/v) phosphoric acid to a pH of 3.2) and methanol (90:10). The flow rate was set at 1.5 mL/min and Dexpanthenol concentration was determined at λmax = 205 nm. The HPLC analysis method was validated in terms of linearity, precision, accuracy, specificity, and sensitivity according to International Conference on Harmonization (ICH) guidelines. The results indicated that the retention time was 8 min and no interferences were observed from the formulation excipients and stress degradation products. Linear regression analysis of data for the calibration plot showed a linear relationship between peak area and concentration over the range of 10 - 100 μg/mL; the regression coefficient was 0.996 and the linear regression equation was y = 20.011 x + 146.83. This HPLC method was precise and accurate in the range of 10 – 100 μg/mL. Also, the dexpanthenol concentration in artificial tear formulation was determined by this HPLC method, which was in accordance with the label claimed. This validated HPLC method could be used for routine analysis, quality control and the stability of analysis of eye gel containing dexpanthenol formulations.

New Validated Stability-Indicating RP-HPLC Method for The Determination of Docetaxel and Its Related Substances

Docetaxel is used for the treatment of castration resistant prostate cancer. Docetaxel is a semi synthetic chemo therapeutic agent extracted from the leaves of Taxus baccata. Two process related substances and four impurities were separated and quantified using Sunfire C18 column (250 x 4.6 mm, 5µm particle size) on gradient mode (40 ± 5°C). A mixture of acetonitrile: 0.01 % Acetic acid was used as mobile phase and a mixture of acetonitrile: methanol (1:1, v/v) was used as diluent and the compounds were monitored at 230 nm. Forced degradation studies were performed and the method was validated as per ICH guidelines.

Development and Validation of a Stability-indicating RP-HPLC Method Using Quality by Design for Estimating Captopril

The applicability of a quality by design framework for the development of a sensitive, simple and selective, stability-indicating reversed-phase high-performance liquid chromatography analytical method for the analysis of captopril was investigated. Design of experiments using a central composite design approach was used for method development. Twenty experimental runs were performed with acetonitrile content ranging between 28 and 36 % v/v, pH from 2.8 to 3.6 and temperature between 22° and 32°. The experimental data obtained was used to derive a quadratic model for the retention time of captopril. The optimized method produced sharp peaks with good resolution (>2) for captopril and the internal standard with retention times of 3.1 and 6.2 min, respectively. The experimental data revealed that acetonitrile content in the mobile phase and pH are significant factors that affect the retention time and resolution of captopril. Normal probability plots revealed that the residual and predicted data fall approximately on a straight line, indicating that the experimental error for these studies was evenly distributed suggesting that the model could be used to navigate the design space. This approach is useful to expedite method development and optimization activities in analytical laboratories.

A New Stability Indicating Validated RP-HPLC Method for Simultaneous Estimation of Escitalopram and Clonazepam in Bulk and Tablet Dosage Form

Present research work describes sensitive, accurate, validated and welldefined stability-indicating RP-HPLC method for the determination of escitalopram (ESC) and clonazepam (CLNZ) in bulk and tablet dosage forms. Chromatographic separation was achieved on a C18 column using mobile phase of a mixture of phosphate buffer and acetonitrile (55: 45 v/v), pH 5.8 with a detection of 231nm. The retention times for Escitalopram and Clonazepam were found to be as 2.01 min, 4.85 min respectively. Linearity was observed in the range 1–200 μg/mL for escitalopram (R2=0.998) and 1.5–150μg/mL for clonazepam (R2 =0.998). The LOD and LOQ were found 1.63 and 3.130μg/mL for ESC whereas 0.099.and 0.301 μg/mL for CLNZ. The drugs under study has been subjected to several stress conditions like acid, alkali, Photolytic, thermal, oxidative conditions and both the drugs are found stable in photolytic, oxidative and thermal conditions but degradation has occurred in acidic and alkaline conditions. Along with this, the method was found to have enough percentage recovery 98.4fr ESC and 101.11 for CLNZ. The developed validated method was found suitable for the routine analysis of ESC and CLNZ in bulk and tablet dosage form.

Development and Validation of a Stability-indicating RP-HPLC Method for the Estimation of Eltrombopag in Bulk and Tablet Formulation

Development and Validation of a Stability-indicating RP-HPLC Method for the Estimation of Eltrombopag in Bulk and Tablet Formulation

UV-Spectrophotometric and Stability Indicating RP-HPLC Methods for the Determination of the Hepatitis C Virus Inhibitor Sofosbuvir in Tablet Dosage Form

A UV-Spectrophotometric and Stability-indicating RP-HPLC methods are developed and validated for the determination of Sofosbuvir (SOF) in tablet dosage form. The RP-HPLC method is performed on the ACE C18 column (250 mm x 4.6 mm, 5 μm) particle size, using buffer solution of pH 5.0 containing 50 mM potassium dihydrogen phosphate: Acetonitrile (60:40 v/v) as the mobile phase at a flow rate of 1.5 mL/min,injection volume 20 μL and UV detection at 260 nm. This method is validated according to ICH and USP requirements for new methods, which include accuracy, precision, selectivity, robustness, ruggedness, linearity and range. A linear range of 50-500 μg/mL with a correlation coefficient of 0.9996 and Rt value of 4.100 min for SOF. The forced degradation studies as acidity, alkalinity, oxidation, heat, thermal and photo degradation are performed according to ICH guidelines, therefore the method can be considered as a stability indicating one. The UV-spectrophotometric method is based on the measurement of the absorbance of SOF at λmax equals to 260 nm. A linear range of 5-50 μg/mL with a correlation coefficient of 0.9995, good accuracy and recovery for laboratory prepared mixture were achieved. The obtained results for the proposed methods are statistically compared with those obtained by the reported methods for SOF using student’s-t and F-ratio tests, showing that the proposed methods are accurate and precise.

A new stability indicating RP-HPLC method development and validation for simultaneous estimation of emtricitabine and tenofovir with degradation kinetics.

A new, simple, rapid and stability-indicating RP-HPLC method was developed for the estimation of Emtricitabine (EMT) and Tenofovir disoproxil fumarate (TDF) in pharmaceutical dosage forms and validated. The HPLC method was developed on SHISEIDO C18 column (250 x 4.6 mm i.d, 5μ) using methanol: 20mM phosphate buffer (pH 5.0) in the ratio of 35: 65 v/v at 271 nm. The retention times for EMT and TDF was found to be 2.79 and 4.91 min respectively. Linearity was established in the range of 0.5–1.5 μg/ml and 0.75–2.0 μg/ml for EMT and TDF respectively. The method was precise with %RSD < 2 for both intraday and interday precision. The accuracy of the method was performed over three levels of concentration and the recovery was in the range of 98–102%. The drugs were individually subjected to forced degradation (thermal, photolytic, hydrolytic, and oxidative stress conditions) studies for seven days. EMT showed maximum degradation in acidic medium and TDF showed maximum degradation in basic medium. The method was successfully applied for quantifying the drugs in marketed dosage forms.

Development and validation of a stability-indicating RP-HPLC method for simultaneous determination of dapagliflozin and saxagliptin in fixed-dose combination

A simple and precise stability indicating method for the simultaneous estimation of dapagliflozin and saxagliptin in combined tablet dosage form was developed and validated using RP-HPLC.

A Validated Stability-indicating RP-HPLC Method for Analysis of Azelaic Acid in Pharmaceuticals

The study is designed to develop a simple, rapid, selective and robust stability indicating reversed phase-high performance liquid chromatography method for quantitative analysis of azelaic acid in pharmaceutical preparations. The chromatographic separation and estimation of azelaic acid was carried out using a Waters high performance liquid chromatography system employing Kromasil 100-5C18 column (250×4.6 mm; 5 µm particle size) as a stationary phase. The mobile phase comprised of 75 volumes of sodium di-hydrogen orthophosphate (pH 3.5; 50 mM) and 25 volumes of acetonitrile, eluted at a flow rate of 1.2 ml/min. The eluents were monitored at 206 nm using 2487 dual wavelength ultra violet detector. The method was developed and validated in terms of stability as per International Conference on Harmonisation and Center for Drug Evaluation and Research guidelines. A linear relationship between peak area and concentration of azelaic acid was observed in a concentration range of 5-400 µg/ml (correlation coefficient, r2= 0.998). The method showed acceptable levels of precision (%RSD ≤2), accuracy (>96 % recovery), robustness (<10 % content difference) and stability (>96 % recovery) over varied environmental and laboratory conditions. The method was successfully applied for the determination of azelaic acid, extracted from three different batches of Aziderm® cream, which yielded a recovery of >97 %, indicated its applicability in routine analysis of azelaic acid in pharmaceutical preparations.

Stability-Indicating RP-HPLC Method for Simultaneous Quantification of Ombitasvir, Paritaprevir and Ritonavir in Tablet Dosage Form

Stability-Indicating RP-HPLC Method for Simultaneous Quantification of Ombitasvir, Paritaprevir and Ritonavir in Tablet Dosage Form

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF LEDIPASVIR AND SOFOSBUVIR IN BULK AND PHARMACEUTICAL DOSAGE FORM

The day by day new combinations drugs are being introduced in market. Then the multiple therapeutic agents which acts at different sites are used in the management of various diseases and disorders are done. Thus it is necessary to develop methods for analysis with the help of number of analytical techniques which are available for the estimation of the drugs in combination. The analyst were determine the Specific, accurate, simple, selective and stability-indicating RP-HPLC method is developed and validated for simultaneous determination of sofosbuvir and ledipasvir in tablet dosage form. RP-HPLC method was performed on the systronics isocratic HPLC System equipped with SP930 D HPLC pump and dual wavelength UV-VIS detector and C18 column (250 mm × 4.6 mm, 5μm), using the mobile phase (Methanol: Water 83:17 v/v) pH 3.0 with 0.05% acidic acid at a flow rate of 1.0 ml/min, injection volume 20μl and UV detection at 245 nm. This method is validated according to BP, USP and ICH requirements for new methods, which include accuracy, precision, robustness, ruggedness, lod, loq, linearity and range. Linear relationships were obtained in the ranges of 10-50μg/ml and 40-200μg/ml with correlation coefficients of 0.9991 and 0.9994 at Rt value of 7.45 min and 3.50 min for sofosbuvir and ledipasvir respectively. The forced degradation studies as acidity, alkalinity, oxidation and hydrolytic degradation were performed according to ICH guidelines.

Stability-indicating RP-HPLC method for simultaneous quantitation of tramadol and aceclofenac in presence of their major degradation products: Method development and validation

ABSTRACTPrimary objective of this study was to develop a stability-indicating reverse-phase high-performance liquid chromatography (HPLC) method for simultaneous quantitation of tramadol and aceclofenac in presence of their degradation products. The drugs were subjected to various International Conference on Harmonization recommended stress conditions, such as acid hydrolysis, alkaline hydrolysis, peroxide oxidation, thermolysis, and photolysis. The major degradation products got well resoluted from the analytes in HPLC analysis with a mobile phase composed of a mixture of 0.01 M ammonium acetate buffer (pH 6.5) and acetonitrile (65:35, v/v) through a Phenomenex Gemini C18 (250 mm × 4.6 mm, 5 µm particle size) column. The method was linear over a range of 15–60 µg/mL for tramadol and 40–160 µg/mL for aceclofenac concentration. The analytes were detected at a wavelength of 270 nm. The method was validated and found to be specific, accurate, precise, stable, and robust for its intended use. The method can be recommended for its future use in routine quality control, accelerated and real-time stability analysis of the formulations containing tramadol and aceclofenac combination.