Stability-indicating Reversed-phase HPLC Research Articles

Stability-indicating Reverse Phase-HPLC Method Development and Method Validation for Quantitative Determination of Degradation Products in Favipiravir API and Drug Product

Introduction: Favipiravir is an antiviral medication shown to be broad spectrum activity against RNA viruses, and potentially treating the COVID-19. Methodology: In this study, the HPLC method for the quantification of degradation impurities (Favipiravir Acid Impurities) were developed and validated for Favipiravir in Tablet dosage form. The specificity of the method was achieved in analytical column Agilent HPLC-C18, 5µm, (4.6 x250) mm. using a suitable mobile phase was 10 mM Phosphate buffer (pH 3.5 with orthophosphoric acid) and Acetonitrile in the Isocratic more of 70:30 v/v. The flow rate is 1.0 mL/min. the injection volume is 10 µL, detection at 320 nm in UV and total run time is 8.0 minutes. The samples were made for forced degradation under hydrolysis, oxidation, photolytic and thermal conditions. The method was validated for specific, selective, linear, robust and accurate as per the ICH guidelines. Results and Conclusion: The linearity of the method for Impurities and the analytes was found from 25 to 150 % concentration level with the correlation coefficient (r2) > 0.999. The accuracy for impurity and the analytes was performed from 50 to 150% level concentration, and mean recovery was found from 98-102%. The analytical degradation and validated study results indicate its unstable nature in acidic, basic and thermal conditions. Therefore, this method is simple, selective and sensitive, this method can be used in pharmaceutical research and development and quality control departments.

A Novel Stability-Indicating RP-HPLC Method for the Simultaneous Estimation and In Vitro and In vivo Evaluation: Curcumin and Naringin Co-amorphous System

Curcumin (CUR) is a phytochemical widely used in food industries, cosmetics, and in the treatment of various ailments. It is a polyphenol derived from turmeric and is often considered the golden spice. CUR has a low solubility of less than 1 µg/ml and poor oral bioavailability which can be improved by co-amorphization with naringin (NRG). Analytical method to simultaneously quantify CUR and NRG is not reported in literature. This study aimed to develop a stability-indicating reverse phase HPLC method in gradient mode to simultaneously quantify CUR and NRG in co-amorphous system. The co-amorphous system of CUR and NRG in molar ratios 1:1 and 1:2 was prepared by quench cooling technique. The separation was attained on a Genesis C18, (4.6 mm × 150 mm, 4 µm) column with the mobile phase comprising of methanol and a 0.1% acetate buffer pH 3.8 at a single wavelength, 289 nm. CUR and NRG eluted at 5.1 and 11.1 min, respectively. For both the molecules, the linearity range was 0.125–16 µg/ml with LOD and LOQ of 0.063 and 0.125 µg/ml. The method developed was validated as per International Conference on Harmonization (ICH) guidelines for linearity, accuracy, precision, and robustness. The method was used to estimate CUR and NRG content in co-amorphous mixture and for in vitro evaluation.Graphical

Analytical Method Development, Validation and Forced Degradation Study of Dapagliflozin by RP-HPLC.

Worldwide, it is projected that 285 million individuals have diabetes, and by 2030, this number is expected to climb to 438 million. About 90% of cases of diabetes mellitus are type 2 (T2DM). Insulin sensitizers, such as metformin and thiazolidinediones; insulin secretagogues, such as sulfonylureas and glinides; dipeptidyl peptidase 4 (DPP-4) inhibitors; glucosidase inhibitors, or oral combination therapy are currently available treatments for type 2 diabetes. Some of these drugs exhibit serious limitations; thus, it is crucial to design an innovative therapy that is efficient and depends on a new channel. In the current work, a stability-indicating reverse phase HPLC (RP-HPLC) technique was developed and subsequently validated for the detection of dapagliflozin in its API. The stability-indicating HPLC method for assay included the use of Kromasil 100-5-C8 (100 mm × 4.6 mm) column, UV detector 224 nm, mobile phase composition involving a mixture of acetonitrile:water (52:48), and a flow rate of 1.0 mL/min. ICH guidelines were followed for the method's validation. To assess the method's specificity and stability in showing characteristics, stress degradation studies were carried out. The working standard solution of dapagliflozin was exposed to 1 and 2 N HCl by refluxing 1 and 2 N NaOH with 30% hydrogen peroxide by volume and UV radiation in order to conduct a degradation study. All system suitability parameters were determined to be within the intended ranges, and the drug's retention duration was discovered to be 1.67 minutes. It was also investigated as to how the drug degraded under various circumstances. The drug was discovered to be stable under situations of photolytic, thermal, neutral, alkaline, and oxidative deterioration. The developed stabilityindicating HPLC technique was validated in accordance with ICH Q2 recommendations, and the validation parameters, such as linearity, precision, and robustness, were achieved within the approved standards. It may be concluded that this method is stability-indicating and specific, and it can be successfully applied to analyze tablet dosage forms containing dapagliflozin.

Greenness profile and whiteness assessment of the stability-indicating HPLC method for the assay of levetiracetam

Adoption of green analytical techniques has been one of the primary goals of the analytical chemistry community over the past years. The reduction of waste generated during sample preparation or analysis is the primary objective of green analytical chemistry (GAC). For the purpose of determining levetiracetam (LEV) in the presence of its alkali-induced degradation product, an accurate, selective, and green stability-indicating reversed phase HPLC technique with UV detection has been devised and validated in the current work. The mobile phase consists of phosphate buffer (pH 3.1) and acetonitrile (87:13 v/v) at the flow rate of 1.0 mL/min, and UV detection was carried out at 210 nm. The stationary phase was Lichrosorb RP-18 (250×4.6 mm i.d., 5 μm particle size) column at 25 °C, in an isocratic mode. The method provided satisfactory resolution and sensitivity, with retention times of (5.6±0.01) minutes for the intact drug and (6.9±0.01) minutes for the alkali-induced degradation product. Different greenness assessment tools, such as the green analytical procedure index (GAPI), Analytical greenness metric (AGREE), and Analytical Eco-Scale (AES), confirmed the proposed approaches' eco-friendliness. The recently launched White Analytical Chemistry (WAC) was also designed to assess the whiteness qualities. The method can be applied for the assay of the cited antiepileptic agent in its formulation in quality control laboratories.

Development of a Simple Extraction Procedure and an HPLC Method for the Determination of Delmopinol in a Challenging Rubbery Dental Chew.

Delmopinol hydrochloride (delmopinol) is widely used in oral hygiene products such as dental chews for preventing dental plaque buildup and gingivitis. This study aimed to develop a simple and inexpensive extraction method, followed by a stability-indicating reversed-phase HPLC (RP-HPLC) method for the determination of delmopinol from rubbery dental chews. The extraction method was optimized in terms of pH, temperature, solvents, repeatability, and reproducibility. Delmopinol was extracted from the rubbery dental chews using 6 N NaOH-MeOH (1 + 1, v/v) at 65°C. The delmopinol peak was eluted in about 7 min by gradient elution on a pH-stable (alkaline) Phenomenex Gemini-NX-C18 column (50 × 4.6 mm id, 110 Å, 3 µm particle size) maintained at 50°C. Mobile phase A was composed of aqueous 10 mM ammonium hydroxide and mobile phase B was acetonitrile. The analyte was monitored by UV detection at 220 nm with a flow rate of 2.0 mL/min. The method was successfully validated as per the current International Conference on Harmonization guidelines for delmopinol with respect to specificity, linearity (R2 >0.999), accuracy, precision, LOQ (2 µg/mL), LOD (0.6 µg/mL), and robustness. The reliability of the method was also demonstrated by batch analysis of 132 dental chews. A simple extraction method and RP-HPLC method were successfully developed and validated for the determination of delmopinol from rubbery dental chews. The overall method was demonstrated to be accurate, robust, specific, and stability-indicating. Therefore, the developed method is suitable and highly desirable for routine analysis of delmopinol in its finished drug products.

Characterization of Degradation Products of Lifitegrast by Mass Spectrometry: Development and Validation of a Stability-indicating Reversed Phase HPLC Method

Lifitegrast API was subjected to forced degradation studies under various conditions of hydrolysis (acidic, alkaline, and neutral/water), oxidation, photolysis, and thermal as prescribed by International Conference on Harmonisation guideline Q1A (R2). A short and simple reversed phase HPLC method was developed. The method was developed on a Shimadzu Ultra C18, (100 x 2.1) mm, 3.0 μm column. The gradient method consisted of 0.1% orthophosphoric acid as mobile phase A and mixture of acetonitrile and methanol in the ratio of 50:50 (v/v) as mobile phase B. The flow rate of the mobile phase was 0.8 mL/min. The developed method was validated using ICH Q2 (R1) guidelines. The parameters considered for method validation were solution stability, specificity, DL/QL, linearity, accuracy, precision and robustness. The drug showed significant degradation in acidic and alkaline conditions while slight degradation was observed in oxidative condition. The drug was found stable in water, photolytic and thermal conditions. The characterization of three major degradation products (DP1, DP2, and DP3) was done with LC-Q-TOF-MS/MS in combination with accurate mass measurements. The most probable mechanisms for the formation of DPs have been proposed on the basis fragmentation pattern.

A novel stability-indicating method for known and unknown impurities profiling for diltiazem hydrochloride pharmaceutical dosage form (tablets)

BackgroundA novel gradient, high-sensitive and specific stability-indicating reverse-phase HPLC method was developed and validated for quantitative purpose of known, unknown and degradant impurities profiling for diltiazem hydrochloride tablets. The impurities were separated on the Zorbax RX C8 column (150 mm × 4.6 mm, 5 μm) with mobile phase-A consisting of a mixture of 0.05 M sodium dihydrogen phosphate monohydrate buffer pH 3.0 and methanol in the ratio 800:200v/v and mobile phase-B consisting of acetonitrile with a flow rate of 1.0 mL min−1. The column compartment was maintained at 35 °C, and the detection wavelength was 240 nm. Diltiazem hydrochloride, its known impurities and unknown impurities have been well resolved from each other.ResultsThe linearity of the method has been demonstrated across the concentration range of 0.18 to 5.65 µg mL−1 for EP impurity-F with correlation coefficient R2 greater than 0.99. Recovery of method was proved from LOQ to 150% for known and unknown impurities with respect to test concentration and found in between 80 and 120%. Forced degradation study and specificity experiment results with mass balance proved the stability-indicating nature of the method and separated all known, unknown impurities and degradants from each other as well as from main drug component (diltiazem hydrochloride). The mass balance for stress study was found in between 95 and 105%.ConclusionNewly developed analytical method was validated as per ICH Q2 (R1) guidelines “Validation of analytical procedure” and found linear, accurate, specific, robust and precise in the established working range.

A Validated Reversed-Phase HPLC Analytical Method for the Analysis of Fenofibrate in Bulk Drug and Tablet Dosage Formulation

Aims: A accurate, precise, and stability-indicating Reversed-Phase HPLC technique has been established for the estimation of fenofibrate in tablet formulation.
 Study Design: Experimental study.
 Place and Duration of Study: Department of Pharmaceutical Sciences, RTM Nagpur University, Nagpur-440033, Maharashtra, India between June 2019 and March 2020.
 Methodology: The chromatographic separation was attained on RP Princeton column (C18) (250 mm x 4.6 mm, 5 µ) with mobile solvent system as a mixture of water (pH 3.0 along o-phosphoric acid) and acetonitrile in the proportion (40:60) v/v, flow rate 1.0 ml per minute, at 240 nm. The retention time of fenofibrate was 3.905 minutes.
 Results: The method demonstrated linearity in the concentration range of 87-232 µg/ml with a coefficient of correlation (r2) of 0.9994. The % RSD was ˂2% and percentage recovery was found to be 99.13-100.74%. The assay of marketed tablet formulations was found to be 99.98%.
 Conclusion: The developed and validated technique as per ICH rules for specificity, accuracy, precision, linearity, and system suitability. Reverse Phase-HPLC technique was utilized to the market formulation.

RP-HPLC stability-indicating method for simultaneous determination of sodium valproate, methylparaben and propylparaben in oral solution

Abstract A new, sensitive, stability-indicating reversed-phase HPLC method was validated and applied for the simultaneous quantitation of sodium valproate and two paraben preservatives; methylparaben, and propylparaben in the liquid dosage form. Stability tests were carried out through exposure of the analyte's solution to stress conditions. Separation of the analytes was achieved on (waters) C18 Column (150 mm × 3.9 mm, 5 μm). A mixture of 0.05 M monobasic potassium phosphate pH 3.5 and acetonitrile (50:50; v/v) was applied at 1.5 ml/min flow rate and UV detection wavelength at 210 nm. The degradation products and the analytes were completely separated. The linearity was performed in the range of 50–150 % from a target concentration of 10 μg/ml propylparaben, 90 μg/ml methylparaben, and 2.88 mg/ml sodium valproate with a coefficient correlation (R2) 1.0 for methylparaben, propylparaben and sodium valproate. The validation results of the suggested method were in a good agreement with ICH guidelines. Application of the proposed method for analysis of liquid dosage forms was successfully carried out in the routine quality control process.

Development and Validation of a Stability-Indicating Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) Method for Identification, Assay, and Estimation of Related Substances of Ivermectin Drug Substance.

Ivermectin is a potent semi-synthetic antiparasitic drug used in veterinary medicine. It is widely used for the treatment of parasites. This study aimed to develop a stability-indicating reversed-phase HPLC (RP-HPLC) method for assay and identification of ivermectin including identification and estimation of its related substances in bulk drug substance batches of ivermectin. Ivermectin and its related substances were separated on an Ascentis Express C18 column (100 mm × 4.6 mm id, 2.7 µm particle size) maintained at 45°C (column temperature) on an HPLC system with gradient elution. The mobile phase was composed of water - acetonitrile (ACN; 50 + 50, v/v) as mobile phase A, and isopropanol - ACN (15 + 85, v/v) as mobile phase B. Analytes were detected with a detection wavelength of 252 nm and quantitated against an external reference standard of ivermectin with a quantitation limit of 0.1% of the target (analytical) concentration. The HPLC method was able to separate all analytes of interest by gradient elution within 25 min. The method was validated according to the guidelines described in the International Conference on Harmonization guideline Q2(R1). The HPLC method for assay of ivermectin and estimation of its related substances was successfully developed, validated, and demonstrated to be accurate, robust, specific, and stability indicating. The performance of the HPLC method is significantly faster and possesses a higher degree of selectivity. Implementation of this method for routine analysis in QC laboratories would save significant time, resources and solvents.

A RP-HPLC Method for the Analysis of Neostigmine Methylsulfate and Process-Related Impurities, Forced Degradation Studies, in the Injection Formulation.

Neostigmine methylsulfate is an anticholinesterase agent and is clinically used for treating myasthenia gravis. It is also used for reversing nondepolarising neuromuscular blocking agents. Neostigmine methylsulfate may be administered by intravenous, intramuscular, or subcutaneous injection. In this research paper, a distinct stability-indicating reverse phase HPLC method was developed and validated for the quantitative determination of related impurities and degradation impurities in neostigmine methylsulfate API and injection formulation. The specific objective was to improve the resolution between European Pharmacopoeia listed impurity A and impurity B and degradation impurity of neostigmine methylsulfate API and injection formulation. The analysis was performed using Kromasil C18 column at 30°C of column oven temperature with phosphate-buffer/acetonitrile in a gradient mode. The RP-HPLC method was developed and validated for in-house neostigmine methylsulfate synthesis process sample and injection formulation. The injection formulation sample was studied for accelerated stability, temperature cycling stability, and photostability. The validation studies for neostigmine methylsulfate synthesis process API were studied using impurity A, impurity B, and impurity C. The analytical method validation parameters studied were specificity, precision, linearity, limit of detection, limit of quantitation, accuracy, and robustness. The API and the injection formulation were subjected to forced degradation under acid, alkali, oxidation, and photolytic and thermal conditions. The proposed method showed a significantly improved RRT (Relative Retention Time) of impurity A and impurity B with a resolution greater than 1.5. The developed method eliminates the use of an ion-pairing agent and thereby a good performance of column was established.

Development and Validation of a Stability-Indicating HPLC Method for Assay of Milbemycin Oxime and Estimation of Its Related Compounds

A stability-indicating reversed-phase HPLC method for assay of milbemycin oxime (MO) and estimation of its related compounds has been developed and validated as per VICH and ICH method validation guidelines. The stability-indicating capability of this method has been demonstrated by adequate separation of all process-related impurities including all potential degradation products of milbemycin oxime (MO) that were generated using stress degradation conditions that are prescribed in ICH guideline. The stability-indicating capabilities of this method were further challenged by analyzing aged stability samples of MO active pharmaceutical ingredient (API). Chromatographic separation of MO and its related compounds was achieved using an isocratic elution at a flow rate of 0.5 mL/min on a Supelco Ascentis Express C18 column (100 mm × 3.0 mm, 2.7 µm particle size, 90 A pore size) maintained at 50 °C. The isocratic mobile phase was composed of 30% v/v of 0.05% phosphoric acid in aqueous solution and 70% v/v of a mixture of methanol and acetonitrile (6:4, v/v). UV detection at 244 nm was employed to monitor the analytes.

Stability Indicating, pH and pKa Dependent HPLC–DAD Method for the Simultaneous Determination of Weakly Ionizable Empagliflozin, Dapagliflozin and Canagliflozin in Pharmaceutical Formulations

A simple, rapid, and accurate stability-indicating reverse phase HPLC–DAD method was developed and validated for the simultaneous determination of empagliflozin, dapagliflozin, and canagliflozin (Gliflozins). Optimum chromatographic separations among gliflozins in the presence of matrices and degradation products have been achieved within 7 min by using Hypercil™ C18 column (25 × 4.6 mm, 5 μm) with acetonitrile and 0.1% formic acid buffer, pH 3.7 (60:40 v/v) as the mobile phase at a flow rate of 1 mL min−1. The proposed method was performed at 230 nm for empagliflozin and dapagliflozin while detection of canagliflozin was carried out at 290 nm. Analytical performance of the method was thoroughly validated in accordance with the ICH guidelines with respect to system suitability, linearity, accuracy, precision, specificity, robustness, detection and quantification limits. Regression analysis showed good correlations with regard to R2 ≥ 0.997 over the concentration range of 4–160 µg mL−1 for gliflozins. The LOD was found to be 0.07 µg mL−1, 0.12 µg mL−1 and 0.29 µg mL−1 for empagliflozin, dapagliflozin and canagliflozin, respectively. Method development was established keeping in view the structure of drugs, their pKa values, ionizability of drugs, pH values of buffer system and buffer capacity. Peak purities of gliflozins in stress tests are less than 1.5, which further confirms no co-elution of degradation products. The proposed method is suitable for routine quality control analysis of gliflozins and to carry out stability studies.

Development and Validation of Stability-indicating Reverse Phase HPLC Assay for Tramadol in Bulk and Tablet Formulations

This manuscript describes the development and validation of a simple, isocratic, and accurate reversed-phase HPLC method for the assay of tramadol in bulk powder form and tablet formulations. The chromato-graphic separation was achieved on an Qualisil BDS C18 (250 mm x 4.6mm, 5μm) column using acetonitrile: methanol: phosphate buffer pH 3.4 adjusted with orthophosphoric acid: (20:10:70, v/v) as a mobile phase, at a flow rate of 1 mL/min and detection at 271 nm. The linear range for tramadol were 2.0 to 10 μg/mL was obtained with correlation coefficients ≥ 0.999. The retention time was found to be 4.47 min for tramadol that was subjected to stress conditions, such as hydrolysis, oxidation, photolysis and thermal degradation, and the stressed samples were analyzed using the above methodology. The method was validated for the precision, accuracy, linearity and robustness. The developed stability-indicating method for tramadol was validated as per ICH guidelines.

A VALIDATED STABILITY INDICATING RP-HPLC METHOD DEVELOPMENT FOR ANTI CANCER DRUG ENZALUTAMIDE IN BULK AND PHARMACEUTICALS

A reproducible stability-indicating Reverse Phase-HPLC technique for the quantification of enzalutamide in in pharmaceuticals was developed and validated. Chromatography was achieved on Inertsil-ODS-C18 (250mm×4.6 mm) 5µmanalytical column with acetonitrile: methanol: water in 40:30:30% v/v proportion as mobile phase and flow rate of 1 ml/min. Enzalutamide was detected at 237 nm UV-wavelength maximum. In the present work mobile phase used as a diluent. Developed technique was validated over 20-150 µg/ml linear concentration range for enzalutamide. This method established with linearity coefficient value of 0.99 and the percentage recovery was found to be 99.3%. This method was proven with LOD and LOQ values of 0.53 µg/mL and 1.61µg/ml respectively. The drug was degraded in acid and alkaline conditions and the percentage degradation values were 3.10 % and 4.54 % respectively. There was no degradation of drug when exposed to neutral, UV, thermal, sun-light and oxidative conditions.Drug was undergoing degradation when exposed to acid and alkaline conditions. The developed technique was useful in the routine quantitation of enzalutamide.

Validation of Stability-Indicating Reverse Phase HPLC Method for the Determination of Related Substances in Dapagliflozin Drug Substance

Validation of Stability-Indicating Reverse Phase HPLC Method for the Determination of Related Substances in Dapagliflozin Drug Substance

Quantitative determination of clobetasone butyrate in bulk and cream formulation by a validated stability-indicating reversed-phase HPLC method

A simple isocratic reversed-phase HPLC method for quantification of clobetasone in bulk and cream dosage forms has been developed. Chromatographic analysis was accomplished on an C18 column utilizing a mixture of methanol and water (84:16 v:v, pH = 6.0) as mobile phase. An effluent flow rate of 1 mL/min was adjusted and the detection was made at 240 nm wavelength. The method was evaluated according to ICH guidelines Q2 R1 for linearity, specificity, sensitivity, precision and accuracy. The method exhibited good linearity with correlation coefficient (r2) of 0.9993 over the concentration range from 5 to 50 μg/mL. The recoveries of the test drug from the cream sample was found to be 98.56 to 99.51% and the limit of detection and quantification were calculated as 0.85 and 2.83 μg/mL, respectively, suggesting the accuracy and sensitivity of the developed method. The precision was demonstrated by a low percentage of relative standard deviation (<1%) from six independent assay analysis performed for the cream formulation. Stability indicating property of the proposed method was demonstrated by performing the analysis of forced degradation samples. The developed method can be used for estimation of the clobetasone butyrate in bulk and pharmaceutical formulations for routine analysis in the quality control laboratories.

Validated Stability-Indicating Chromatographic Methods for the Determination of Chlordiazepoxide and Clidinium Bromide in the Presence of its Alkali-Induced Degradation Product: Kinetic Study

Accurate, simple and selective stability-indicating reversed phase HPLC and TLC-densitometric methods with UV detection have been developed and validated for simultaneous determination of chlordiazepoxide (CDZ) and clidinium bromide (CDB) in the presence of its alkali-induced degradation product (DEG). Successful separation of the drugs from the degradation product was achieved. For the RP-HPLC method, the stationary phase was ACE-126-2546 AQ C-18 (250 × 4.6 mm i.d., 5 μm particle size) column at 25 °C, in an isocratic mode, using mobile phase containing a mixture of 25 mM ammonium acetate (pH 5.4): acetonitrile in the ratio of (20:80, v/v), at the flow rate of 1.0 mL min−1 and UV detection was performed at 222 nm. The retention times were (4.13 ± 0.01) and (8.5 ± 0.01) min for CDZ and CDB, respectively. For the TLC-densitometric method, the separation was performed using a stationary phase of precoated Silica Gel G/UV254 and mobile phase composed of a mixture of ethyl acetate: methanol: ammonia (8:3:1, v/v/v) and scanned at 222 nm. The R f values were (0.79 ± 0.02) and (0.11 ± 0.01) for CDZ and CDB, respectively. The linearity graphs for CDZ and CDB, respectively, were found to be linear over (0.5–40) μg mL−1 and (2–45) μg mL−1 with mean percentage recoveries (99.69 ± 0.836) and (99.28 ± 1.838) for RP-HPLC method and (1–14) μg band−1 and (0.5–10) μg band−1 with mean percentage recoveries (100.00 ± 0.782) and (100.19 ± 1.010) for TLC-densitometric method. A comparative study of different analytical validation parameters such as accuracy, precision, specificity and robustness were conducted. The obtained results were statistically compared with those of the official and reported methods; using Student’s t test, F test and one-way ANOVA, showing no significant difference with high accuracy and good precision. The proposed RP-HPLC method was also used to study the kinetics of the alkaline hydrolysis of clidinium bromide that was found to follow pseudo-first order kinetics. The t 1/2 was 8.5729 min while k (the degradation rate constant) was 0.0808353 min−1.

Development and validation of a novel RP-HPLC method for stability-indicating assay of Abiraterone acetate

ABSTRACTAbiraterone acetate is a prodrug of Abiraterone widely used for the treatment of metastatic castration resistant prostate cancer. In this study, a simple, sensitive, and rapid stability-indicating reverse phase HPLC method was developed and validated for the determination of Abiraterone acetate in bulk and its pharmaceutical formulation. The method was developed by HPLC using a Hypersil ODS C-18 (150 mm × 4.6 mm, 5 µm) column in a isocratic mode with mobile phase constituted by potassium phosphate buffer and acetonitrile (40:60, v/v%) flow rate was 1.0 mL min−1, column temperature of 30°C, UV detection wavelength 235 nm, and injection volume of 20 µL. The validated parameters were in accordance with FDA and ICH specifications, assay exhibited a linear range of 25–250 µg mL−1 with regression (r2) coefficient 0.9998. The limits of detection and quantification were 0.23 and 0.70 µg mL. Accuracy was between 99.34 and 100.07%. The drug was subjected to various stress conditions like acidic, base hydrolysis, oxidation, thermal, and photolytic degradation. Stress study Abiraterone acetate was found susceptible to degrade under hydrolytic (acid and base) conditions. The proposed method has stability indicating the resolution of the main peak from their degradation peaks. The validated method is suitable for quality control application and reduced analysis time.

A stability indicating RP-HPLC assay method for simultaneous estimation of abacavir, Lamivudine, Nevirapine and Zidovudine in pharmaceutical dosage form

A stability-indicating reverse phase HPLC method has been developed and validated for the simultaneous Assay estimation of Abacavir, Lamivudine, Nevirapine and Zidovudine in pharmaceutical dosage form. The chromatographic separation was achieved on a simple Inertsil ODS C18, 250 x 4.6 mm, 5 µm (Make: GL Sciences). The method employed a simple isocratic elution mode with a ternary mixture of mobile phase. The detection wavelength was set at 270 nm. The proposed method was extensively validated as per ICH guidelines. The specificity of the method was proved by performing forced degradation studies along with peak purity. This method is found to be simple, fast and economical Hence this validated method can be used in routine quality testing of individual dosage forms and combination dosage forms of Abacavir, Lamivudine, Nevirapine and Zidovudine.