Stability-indicating RP-HPLC Method Research Articles

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.

Development and Validation of Stability-Indicating RP-HPLC Method for the Estimation of Cuminaldehyde

Cumin aldehyde is an herbaceous plant (Cuminumcyminum L.) volatile oil that used as a regular spice in kitchen foods. It also has some pharmacological properties such as analgesic, hepato-protective, anti-inflammatory, antibacterial, antimicrobial, antifungal, antiviral, antioxidant and anticancer. The key objective of the present study was to develop and validate the RP-HPLC method for the estimation of cumin aldehyde. According to the ICH guidelines, RP-HPLC stability indicating method was used in which reverse phase enable Cosmosil C18 column (250 × 4.6 mm, 5μm) used in isocratic mode. For the estimation of cumin aldehyde, sodium sulphate: acetonitrile: methanol (20:73:7 v/v) was used as a mobile phase and it was delivered at flow rate of 1.0 mL min- 1. The injection volume was 20 µL and elute was analyzed by a UV detector at 326 nm. Linearity was observed between the concentration range of 20 μg mL-1 -140 μg mL-1 and the correlation coefficient R2 value was found to be 0.997 ± 0.5. The method was accurate and recovery was found to be in the range of 98.06 -100.40 %. The limit of detection of cumin aldehyde was found to be 1.04 μgmL-1and limit of quantitation was found to be 3.16 μgmL-1. Cumin aldehyde was subjected to stress conditions including acidic, alkaline, neutral, oxidation, and dry heat degradation. Cumin aldehyde was more sensitive to acidic, dry heat and oxidative degradation and it is stable at alkaline conditions. The method was validated according to ICH guidelines.

Quantification of Clofarabine and its Impurity Substances by RP-HPLC Method in Parenteral Formulation

&#x0D; A simple, accurate and precise stability-indicating RP-HPLC method was developed for estimation of clofarabine in parenteral formulation. Some known impurities like, alpha-anomer, 2-chloroadenine and monobenzoate are used which are related substances of clofarabine, for the analysis of the marketed formulation. Inertsil C18 (150 mm × 4.6 mm) 5μ (particle size) was used as stationary phase and Buffer: Acetonitrile 90:10 v/v was used as mobile phase with flow rate 1 ml/min at 263 nm UV detection. The retention time of clofarabine was found to be 3.07 minutes. Linearity was observed over the concentration range of 5-25 μg/ml for clofarabine in which correlation coefficient value was found to be 0.999. Force degradation study was perform and maximum degradation of Standard and Test of Clofarabine was found to be 18.8% and 17.5% respectively in Acidic condition. The LOD and LOQ was found to be 0.071 μg/mL and 0.21 μg/mL, respectively for clofarabine. Moreover, the % RSD for repeatability, inter and intra day precision was found to be within the range, which reveals that the method is precise. Accuracy study of the drug in marketed preparation also reported within the limits. In conclusion, the assay is useful for estimation of clofarabine in parenteral formulations.

User-Friendly HPLC Method Development and Validation for Determination of Enalapril Maleate and Its Impurities in Enalapril Tablets.

The official method for the determination of Enalapril Maleate and its related substances in European Pharmacopoeia (EP) is a gradient liquid chromatographic method. The method used styrene-divinylbenzene copolymer column, mobile phase buffer pH 6.8 and column oven temperature 70°C. In this method, the separation between main component Enalapril and Ph. Eur. Imp-A was not completed hence the achieving system suitability requirement is a tough task and it requires quite often adjustment in chromatographic parameters. Moreover, column oven temperature 70°C is not user friendly to HPLC instruments and users. In this study, several changes were introduced to the method in order to improve the separation, peak shapes and to overcome the column oven temperature. A new user-friendly stability-indicating RP-HPLC method was developed for Enalapril related substances analysis. The developed method uses a ZORBAX Eclipse XDB-C18 column with column oven temperature at 55°C and mobile phase containing acetonitrile and a phosphate buffer at pH 3.0. The method is capable of separating all the known impurities with resolution more than 3.5, which is much better than that obtained with the existing monograph methods. The optimized method was validated and demonstrated to have acceptable specificity, sensitivity, linearity, accuracy, precision, robustness, solution stability and equivalency to the EP method. The developed method proved to be applicable to a wide number of C18 reversed-phase columns. In addition, the Enalapril assay method also presented with 20 min run time.

Stability-indicating method for the determination of assay and quantification of impurities in amlodipine–atorvastatin combination dosage form by RP-HPLC

ABSTRACTA simple stability-indicating RP-HPLC method was developed and validated for quantification of amlodipine, atorvastatin, and its impurities on Waters HPLC using Unisol C18 5 µm, 250 × 4.6 mm column in their combined tablet dosage as per ICH guidelines. The gradient (T/%B) at 0/42, 18/42, 22/75, 30/75, 32/42, and 35/42 of 40 mM 4.7 pH ammonium acetate as mobile phase A and acetonitrile as mobile phase B of flow rate 1.5 mL/min and 240 nm wavelength. Peak purity compiled for amlodipine and atorvastatin in all stressed conditions. For impurities: Precision was found in between 1.5 and 3.6%. The limit of detection and quantification for amlodipine, amlodipine impurity A, and atorvastatin was found to be 0.06 and 0.18 µg/mL, for atorvastatin Impurity A, B, C, and H was determined as 0.04 and 0.11 µg/mL, for Atorvastatin Impurity D was measured as 0.11 and 0.28 µg/mL, respectively. The linear regression achieved >0.9999 from 0.22 to 7.5 µg/mL. Recovery was observed in between 97 and 101%. For assay: Precision was determined in between 0.1 and 0.2%. The linear regression achieved >0.9999 for amlodipine and atorvastatin. Recovery ranged from 100 to 101%. The validated method was found to be accurate, precise, reliable, and robust to determine the assay as well as impurities in amlodipine–atorvastatin combination dosage formulation.

Validated Stability-indicating RP-HPLC Method for the Determination of Parecoxib Sodium and Degradation Impurities in Bulk Drug

Background: Parecoxib sodium (PCX), the selective cyclooxygenase (COX)-2 inhibitor, has aroused great interests among researchers mainly because of its central role in analgesic and antiinflammatory effects in a wide range of perioperative or postoperative procedures. The aim of the current study was to develop and validate a precise, accurate, and specific HPLC method systematically which could accomplish the stability indicating of PCX bulk drug and qualitation and quantization for the formed main impurity under the stressed conditions. Keywords: HPLC-UV/DAD, parecoxib sodium, stability-indicating, degradation products, validation, titrimetric method.

A Simple and Specific Stability- Indicating RP-HPLC Method for Routine Assay of Adefovir Dipivoxil in Bulk and Tablet Dosage Form.

A simple and reliable stability-indicating RP-HPLC method was developed and validated for analysis of adefovir dipivoxil (ADV).The chromatographic separation was performed on a C18 column using a mixture of acetonitrile-citrate buffer (10 mM at pH 5.2) 36:64 (%v/v) as mobile phase, at a flow rate of 1.5 mL/min. Detection was carried out at 260 nm and a sharp peak was obtained for ADV at a retention time of 5.8 ± 0.01 min. No interferences were observed from its stress degradation products. The method was validated according to the international guidelines. Linear regression analysis of data for the calibration plot showed a linear relationship between peak area and concentration over the range of 0.5-16 μg/mL; the regression coefficient was 0.9999and the linear regression equation was y = 24844x-2941.3. The detection (LOD) and quantification (LOQ) limits were 0.12 and 0.35 μg/mL, respectively. The results proved the method was fast (analysis time less than 7 min), precise, reproducible, and accurate for analysis of ADV over a wide range of concentration. The proposed specific method was used for routine quantification of ADV in pharmaceutical bulk and a tablet dosage form.

Development and Validation of a Stability Indicating RP-HPLC Method for the Determination of Imatinib Mesylate

The aim of this paper was to develop and validate the stability indicating RP-HPLC method for the determination of Imatinib mesylate in bulk and pharmaceutical dosage forms. A simple, accurate, precise, sensitive and stability indicating RP-HPLC method has been developed for the determination of Imatinib mesylate in bulk drug and pharmaceutical dosage form, in which separations are done using develosil C18, 5μm, 150 × 4.6mm i.d. column at a flow rate of 1.0mL/min with an injection volume of 20μL. The beer’s law was obeyed over the concentration range of 5 - 35μg/mL. The correlation coefficient was found to be 0.996 and it showed good linearity, reproducibility, precision in this concentration range. The % recovery values were found to be within the limits, which showed that the method was accurate. The LOD and LOQ were calculated using statistical methods. The % RSD values were less than 2. The developed method was successfully applied for determination of Imatinib mesylate in pharmaceutical dosage form. The results obtained are in good agreement with those obtained by using the standard method.

New RP-HPLC Method Development and Validation for the Determination of Pazopanib Hydrochloride in Tablet Dosage Form

The aim of this paper was to develop and validate the stability indicating RP-HPLC method for the determination of Pazopanib hydrochloride in bulk and pharmaceutical dosage forms. A simple, accurate, precise, sensitive and stability indicating RP-HPLC method has been developed for the determination of Pazopanib hydrochloride in bulk drug and pharmaceutical dosage form, in which separations are done using develosil C18, 5μm, 150 × 4.6mm i.d. column at a flow rate of 1.0mL/min with an injection volume of 20μL. The beer’s law was obeyed over the concentration range of 5 - 35μg/mL. The correlation coefficient was found to be 0.996 and it showed good linearity, reproducibility, precision in this concentration range. The % recovery values were found to be within the limits, which showed that the method was accurate. The LOD and LOQ were calculated using statistical methods. The % RSD values were less than 2. The developed method was successfully applied for determination of Pazopanib hydrochloride in pharmaceutical dosage form. The results obtained are in good agreement with those obtained by using the standard method.