RP-HPLC Method For Simultaneous Estimation Research Articles

Development and validation of RP-HPLC method for simultaneous estimation of 5-fluorouracil and silibinin in corn-derived protein-polysaccharide nanocomplex: Forced degradation and mathematical modeling of drug release

In the current work, RP-HPLC method was developed and validated for simultaneous analysis of 5-Fluorouracil (5-FU) and silibinin from corn-derived protein polysaccharide nanocomplex. The method employs a mobile phase of water (A) and acetonitrile: methanol (60:40) (B) on a Waters C18 column (4.6 x 150 mm, 5 μm), with detection wavelengths set at 266 nm and 288 nm for 5-FU and silibinin, respectively. The runtime was 15 min at a flow rate of 0.7 mL/min. The analytical method validation adhered closely to ICH guidelines, exhibiting remarkable selectivity, specificity, linearity, and precision, all falling comfortably within acceptable thresholds. The analytical method was found to be linear from 1 to 150 µg/mL with R2 of 0.9998. Forced degradation studies were carried to assess the stability of 5-FU and silibinin within the nanocomplex across diverse conditions. Additionally, nanocomplexes loaded with 5-FU and silibinin were formulated and characterized, demonstrating a particle size of 209.83 ± 3.83 nm and a zeta potential of 57.93 ± 1.06 mV. The %entrapment efficiency for 5-FU and silibinin was found to be 88.73 ± 0.37 % and 97.00 ± 1.58 % with a %drug loading of 6.73 ± 0.92 % and 2.74 ± 0.61 % respectively. The %in vitro drug release was determined using developed and validated analytical methods. Further, various release kinetic models were applied to determine the release behavior from the nanocomplex system. To conclude, we ensured the eco-friendliness of the developed analytical method by assessing its greenness through Eco-scale, the green analytical procedure index (GAPI), and the Analytical GREEnness metric (AGREE).

METHOD DEVELOPMENT AND VALIDATION OF RAPID ISOCRATIC RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF PARACETAMOL, CAFFEINE, AND PROPYPHENAZONE IN PHARMACEUTICAL FORMULATION

Objective: The objective is to develop a novel, rapid, simple, precise, accurate, and reproducible RP-HPLC method for simultaneous estimation of paracetamol, caffeine, and propyphenazone in bulk and a pharmaceutical dosage form. Method: Optimized chromatographic conditions were used for isocratic elution with Shimadzu C18 (4.6×250 mm, 5 μm), methanol, and 20mM phosphate buffer (60:40, v/v, pH 2.5) as mobile phase, flow rate 1.2 mL/min, and UV detector at λ max 272 nm. The method was validated for specificity, precision, linearity, accuracy, robustness, and solution stability as per the International Council for Harmonization (ICH) guidelines. Result: It was discovered that the retention time for paracetamol, caffeine, and propyphenazone was found to be 2.6 min, 3.0 min, and 7.5 min. The method proved to be rapid, simple, linear (R2>0.999), precise {relative standard deviation (RSD<2.0%)}, accurate (recovery 98–102%), sensitive, and robust. Conclusion: The proposed novel isocratic RP-HPLC method was found to be rapid (short run time of 10 min), highly selective, accurate, and sensitive. The method has been successfully applied to the simultaneous analysis of paracetamol, caffeine, and propyphenazone in a pharmaceutical formulation.

Analytical Method Development and Validation of Beclomethasone Dipropionate and Fusidic Acid in pure and Pharmaceutical Dosage Form (Cream) by RP-HPLC

A simple, rapid, specific, cheap, eco-friendly, and accurate RP-HPLC method for simultaneous estimation of Beclomethasone Dipropionate and Fusidic Acid in pure and combination drugs has been developed and validated. A stationary phase of BDS Hypersil C18 ((250mm x 4.6m, 5m) and a mobile phase of Acetonitrile: Methanol: Orthophosphoric acid (60:20:20v/v) were used for the separation. The flow rate was continued at 1.0 ml/min and the effluent was observed at 230nm. The designated situations were appropriate for separating Beclomethasone Dipropionate and Fusidic acid eluted with a mean retention time of 7.2 and 5.1 minutes, respectively. The planned technique was validated as per the International Council for Harmonisation recommendation. Its linearity, specificity, system suitability, accuracy and precision, limit of detection, limit of quantitation, and robustness are some of the parameters that have been validated. The concentration range produced linear calibration curves of 1-6g/ml and 2-12g/ml for Beclomethasone Dipropionate and Fusidic Acid with R2 = 0.9961 and 0.9963. Percent RSD values for both intraday and interday were determined to be less than 2.0%. Demonstrating the accuracy of the approach. The technique was proven to be reliable. A successful application of the developed RP-HPLC method for the quantification of Beclomethasone Dipropionate and Fusidic Acid in marketed semi-solid dosage form (Cream).

A Review on Method Development and Validation of RP-HPLC Method for Simultaneous Estimation of Teneligliptin, Metformin Hydrochloride and Pioglitazone Hydrochloride in Pharmaceutical Dosage Form

A Review on Method Development and Validation of RP-HPLC Method for Simultaneous Estimation of Teneligliptin, Metformin Hydrochloride and Pioglitazone Hydrochloride in Pharmaceutical Dosage Form

Development and validation of stability indicating RP-HPLC method for simultaneous estimation of silodosin and mirabegron in synthetic mixture

The study focuses to validate and develop a precise, simple and accurate stability indicating RP-HPLC method for estimation simultaneously of and silodosin and mirabegron in synthetic mixture. The chromatographic separation was achieved by using Shimpack Solar C18 column (250×4.5mm, 5μm) with acetonitrile: 5mM ammonium acetate in ratio of 90:10% v/v as a mobile phase at a constant flow rate of about 1.2mL/min. The development and validation were carried out at detection wavelength of 229nm. We developed a robust RP-HPLC method, validated for linearity, precision, accuracy, specificity, and system suitability. The method demonstrated excellent linearity with correlation coefficient value r2 was nearly 0.998 with linearity range 8-18μg/mL for Silodosin and 24-54μg/mL for mirabegron. LOD and LOQ were found to be lower; hence, the method is sensitive. Percentage recovery was obtained 99.97% and 99.99% for silodosin and mirabegron, respectively. In case of precision, robustness and repeatability, RSD was found to be less than 2. The validated and developed RP-HPLC method offers an efficient and practical approach for the simultaneous quantification of silodosin and mirabegron in pharmaceutical formulations, making it a valuable tool for quality control and pharmaceutical research.

Simultaneous estimation of emtricitabine, tenofovirdisoproxilfumarate and rilpivirine in pharmaceutical dosage form by using rp-hplc

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.

Multipurpose RP-HPLC Method for Simultaneous Estimation of Fixed-Dose Combinations of Anti-diabetic Drugs: Integrating Green, Economical, and Robust Approaches with Design of Experiments and White Analytical Chemistry

Multipurpose RP-HPLC Method for Simultaneous Estimation of Fixed-Dose Combinations of Anti-diabetic Drugs: Integrating Green, Economical, and Robust Approaches with Design of Experiments and White Analytical Chemistry

Development and Validation of an Analytical RP-HPLC Method for Simultaneous Estimation of Losartan and its Active Metabolite (EXP-3174) in Isolated Perfused Rat Liver

Background: The liver perfusion method is frequently used in drug pharmacokinetic studies and the various effects of drugs on liver tissue. The aim of this study was to establish and validate an analytical method using high-performance liquid chromatography to determine the simultaneous concentration of losartan and its active metabolite, EXP-3174, in an isolated perfused rat liver study. Method: An HPLC system with isocratic mode was used. Various chromatographic parameters were adjusted to develop and validate a method for determination of losartan and its active metabolite in liver perfusion media. Results: In this study, losartan and its active metabolite, EXP-3174, were separated using a C18 stationary phase, a mobile phase consisting of acetonitrile: phosphate buffer at a flow rate of 1 mL.min-1, and UV detection at 254 nm. Retention times for losartan and the metabolite were 10 and 16 minutes, respectively. Linearity from 25-250 ng.ml-1 was validated with acceptable accuracy and precision. The LOD and LOQ for losartan were 7.0 and 21.1 ng.ml-1, respectively. The LOD and LOQ for metabolite were 7.4 and 22.4 ng.ml-1, respectively. ChromGate® software was used to acquire and process the data. Conclusions: The optimized and validated technique was effectively used to analyze losartan and its active metabolite in isolated perfused rat liver.

A Validated RP-HPLC Method for Simultaneous Estimation of Benidipine and Telmisartan in Bulk and Tablet Dosage Form

Background: In this work, a novel, simple and accurate reverse phase high-performance liquid chromatographic method with UV detection is presented for the simultaneous estimation of Benidipineand Telmisartan in API and marketed formulations. Objective: To develop and validate this method in accordance to regulatory guidelines i.e. ICH Q2B-R1 for analytical parameters. Methods: In RP-HPLC the separation was achieved using a Xbridge C18 (250x 4.6 i.d; 5µm) column with isocratic elution using a mobile phase consisting of methanol: acetonitrile (80:20v/v/ with 0.1% v/v TEA) adjusted to pH 5.2 with glacial acetic acid at 1mL/min flow for 10min. Benidipine and Telmisartan, were detected at dual wavelength at 237 and 297nm with retention time (Rt) 6.891 and 8.997 minutes respectively. Results: Our new methods (RP-HPLC) were found to (r2>0.999) for both Benidipine and Telmisartan. The developed methods are highly selective, precise and reproducible with %RSD < 2%, Moreover the methods are accurate with recovery more than 99% in both methods. Conclusion: The RP-HPLC method results are efficient, simple and easy. These highly sensitive methods were successfully applied for assay determination of both Benidipine and Telmisartan at fixed dose combination.

Simultaneous Estimation of Eugenol and Scopoletin from the in House <i>Avipattikar Churna</i> by RP-HPLC Method and Estimation of Scopoletin from Different Extracts of <i>Jalap</i>

Background: Although standardising polyherbal medicine requires immediate attention, it is a tedious undertaking. Phytochemical profiling is a particularly useful tool for assessing the quality and effectiveness of polyherbal medicines, among various methods used for standardization. The proposal aimed to develop a precise RP-HPLC method for simultaneous estimation of eugenol and scopoletin in in-house Avipattikar churna. This method was also used to estimate scopoletin in various extracts of Ipomoea turpethum. Methods: The phytomarkers in Avipattikar churna, hydroalcoholic and alcoholic extracts of Jalap were estimated by RP-HPLC system. In this setup, RP-ODS C8 column was employed with methanol: water (30:70 v/v, 0.1% formic acid) at 1 ml/min for 0-10 minutes, and then with methanol: water (60:40 v/v) at 0.8 ml/min for 10.01-25 minutes. Detection was done at 280 nm for eugenol and 366 nm for scopoletin using a UV/VIS detector. The method was validated by performing validation parameters as per ICH guidelines. Results: The linearity of eugenol and scopoletin was performed, with correlation coefficients of 0.999 and 0.9969 respectively. In repeatability, % RSD was observed as 0.856 and 0.909 for eugenol and scopoletin correspondingly. The LOD (detection limit) of eugenol was 0.67 μg/mL and of scopoletin was 1.39 μg/mL. While LOQ (quantification limit) of eugenol was found as 2.04 μg/mL and 4.03 μg/mL for scopoletin. The % recovery was ranging from 102.96 - 100.45 % for eugenol and from 102.65 - 101.3 %w/w for scopoletin, after adding a pre-quantified amount (20 μg/mL) in the different concentrations of the standards. The eugenol and scopoletin were estimated 0.1366 %w/w and 0.0465 %w/w respectively in Avipattikar churna. The hydro alcoholic extract of Jalap showed presence of more scopoletin than in the alcoholic extract. Conclusion: The validated process is established as accurate, consistent and precise which results as a better standardization drive for Ayurvedic dosage forms.

Development and Validation of RP-HPLC Method for Simultaneous Estimation of Gallic Acid, Scopoletin, Umbelliferone, and Imperatorin in Bael avaleha

Bael avaleha is a semisolid ayurvedic formulation containing the fruit of bael, which acts as an anthelmintic to treat diarrhea, dysentery, and IBS. It strengthens the gastrointestinal tract, heals ulcers, and alleviates abdominal pain. The avleha contains excess sugar, which can result in faster absorption of active ingredients in the biological system. Researchers have begun scrutinizing ayurvedic formulations for quality and consistency, with quality testing being conducted to ensure that the quality of ayurvedic formulations adheres to ayurvedic standards. Phytochemical fingerprinting by spectroscopic and chromatographic techniques is becoming more popular in assessing the efficacy of ayurvedic products. Previous studies evaluated phenolic compounds, flavonoids, monoterpenes, and sesquiterpenes in Bael fruit using Gas Chromatography Mass Spectrometry (GC-MS), HPTLC, and HPLC methods.This study evaluated marketed bael avaleha formulations (BA I and II) for organoleptic characteristics, physical parameters, and phytoconstituents. A UV-vis spectrophotometric study was conducted for quantitative analysis of phytochemicals. The extract of bael avaleha formulations BA I and BA II were found to have a total phenolic content of 42.34 ± 0.090 mg GAE/g and 38.52 ± 0.065 mg GAE/g, a flavonoid content of 14.34 ± 0.070 mg QE/g and 12.42 ± 0.086 mg QE/g, sugar content of 69.44 ± 0.020 mg Glu/g and 67.18 ± 0.065 mg Glu/g, and a reducing sugar content of 31.34 ± 0.025 mg Glu/g and 29.08 ± 0.047 mg Glu/g, respectively. The Fourier transform infrared (FTIR) spectra of extract of bael avaleha formulations were recorded in region 4000–400 cm−1 and confirmed the presence of hydroxy group, aromatic C-H stretch, C=C and C-O group. RP-HPLC method was developed and used for the estimation of gallic acid, scopoletin, umbelliferone, and imperatorin in the extract of Bael avaleha formulations

Simultaneous Chromatographic Estimation of Vildagliptin and Dapagliflozin Using Hybrid Principles of White Analytical Chemistry and Analytical Quality by Design.

The fixed-dose combination of vildagliptin (VDG) and dapagliflozin (DGZ) is used in the treatment of type 2 diabetes mellitus. According to the literature survey, RP-HPLC and HPTLC methods have been reported for routine analysis of VDG and DGZ. These chromatographic methods have been developed using potentially neurotoxic and teratogenic solvents, which are unsafe for human and aquatic animal life and hazardous to the environment. These types of organic solvents shall be replaced or reduced during chromatographic analysis of drugs for the safety of human and aquatic animal life and the protection of the environment. The novel white analytical chemistry (WAC) approach has been introduced, which emphasizes robust, green, user-friendly, economical, and rapid analysis of drug samples. Hence, the WAC-based RP-HPLC method has been developed for the estimation of VDG and DGZ using lower toxic and economical solvents. The development of the RP-HPLC method includes the implementation of the analytical quality by design approach using principles of design of experiments to reduce organic waste generation and regulatory compliance of analytical method. The central composite design was applied for response surface modeling (RSM) and optimization of the RP-HPLC method. The method validation was carried out according to ICH Q2 (R1) guidelines. The fixed-dose combinations of VDG and DGZ were assayed, and results were found in compliance with their labeled claim. The published and proposed RP-HPLC methods were assessed for chromatographic analysis of VDG and DGZ using the Red-Green-Blue (RGB) model, AGREE calculator, Eco-Scale Assessment tool, GAPI software, and NEMI standards. The proposed method was found to be robust, green, economical, and user-friendly for chromatographic analysis of VDG and DGZ. The proposed method can be an economical and eco-friendly analytical tool in the pharmaceutical industry for quality control and routine analysis of fixed-dose combinations of VDG and DGZ. Hybrid principles of WAC and analytical quality by design to RP-HPLC method for simultaneous estimation of VDG and DGZ in their fixed-dose combinations.

A Novel RP-HPLC Method for Simultaneous Estimation of Vilanterol Trifenatate, Umeclidinium Bromide and Fluticasone Furoate in Inhalation Dry Powder Formulation.

The dry powder inhalation formulation containing vilanterol trifenatate, umeclidinium bromide and fluticasone furoate intended for the therapy of bronchospasm related to chronic obstructive pulmonary disease and bronchial asthma was selected for the development and validation of a novel, selective, accurate, precise, quick and cost-efficient reversed-phase, high-performance liquid chromatography method. Neither an official monograph nor a single method has yet been published for the simultaneous estimation of these three compounds, which makes this method novel. The stationary phase of an ACE-C18-PFP column (250mm × 4.6mm, 5μ) was used with a mobile phase of 25-mM sodium perchlorate buffer (pH2.5 adjusted with ortho-phosphoric acid) and acetonitrile (40:60% v/v) at a flow rate of 1mL/min to optimize chromatographic variables. The column temperature was kept at 40°C, and detection was at 224nm, which was the isosbestic point of these three drugs. Well-resolved good peak symmetry was obtained for all three molecules by isocratic elution in less than 10min, and the retention times of vilanterol trifenatate, umeclidinium bromide and fluticasone furoate were found to be 3.7, 5.4 and 8.3min, respectively. The proposed method was validated as per ICH Q2 (R1) guidelines, and the calibration curves were linear in concentration ranges of 5-35μg/mL for vilanterol trifenatate, 5-80μg/mL for umeclidinium bromide and 5-150μg/mL for fluticasone furoate, with mean % recoveries of 99-100%. The limits of detection and quantitation are 0.15 and 0.45μg/mL for vilanterol trifenatate, 0.58 and 1.77μg/mL for umeclidinium bromide and 0.32 and 0.96μg/mL for fluticasone furoate, respectively. Hence, the proposed RP-HPLC technique was successfully used to quantify the inhalation formulation containing all three compounds.

Development and Validation of Chemometric Assisted RP-HPLC Method for Simultaneous Estimation of Perindopril Erbumine, Indapamide, and Amlodipine Besylate in Bulk and Pharmaceutical Formulation

This research paper illustrates a latterly developed, optimized and validated gradient RP-HPLC approach for simultaneous analysis ofIndapamide, Perindopril erbumine and Amlodipine besylate in bulk and pharmaceutical formulation with the assistance of quality by design. Qualityis predicated on desired and predetermined specifications. Understanding various factors, dependent variables, and their interconnection effectsby a desired set of experiments on the responses to be analyzed is an important component of QbD. Several operating conditions of variousprocesses optimization, chromatographic separation performance improvement, and high extraction efficiency were attained by using QbD. Thepowerful chromatographic conditions were done using the HypersilC18 column (250mm × 4.6mm, 5μm particle Size). The UV detector wasadjusted to 215nm. Design of experiments (DoE) was applied for multivariate optimization of the experimental conditions of the RP-HPLCmethod. Three independent factors, mobile phase composition, phosphate buffer strength, and flow rate, were used to design mathematicalmodels. Central composite design (CCD) was used to examine the response surface methodology and fully examine the results of theseindependent factors. The desirability function was used to optimize the retention time and resolution of the analytes simultaneously. Theimproved and anticipated data from the contour diagram consisted of methanol and phosphate buffer (pH 2.5, strength 0.05M) in the ratio of65:35, respectively, at a flow rate of 1.1 ml/min. Using these optimum conditions, baseline separation of both drugs with good resolution and runtime of less than 5.0 min was achieved. The novelty of the developed method was time-consuming, cost-effective, and sensitive. The optimizedassay conditions were validated according to ICH guidelines. Under the optimized state, the linearity ranges were found to be 10-40 μg/mL, 32–128 μg/mL, and 40-160 μg/mL for Indapamide, Perindopril erbumine, and amlodipine besylate, respectively, with correlation coefficients (R2) of0.999. The mean accuracy studied ranged from 99.18 to 99.58%. The percentage coefficient variation value for the precision study was lower than1%. The proposed method showed good precision and repeatability. Hence the developed RP-HPLC method using quality by design can be usedas a routine quality control analysis of indapamide, perindopril erbumine, and amlodipine besylate.

Development and validation of RP-HPLC method for simultaneous estimation of Ertugliflozin and Sitagliptin in bulk drug and tablet dosage form

To treat type 2 diabetes, in a combined tablet dosage form the ertugliflozin and sitagliptin were administered. Considering the less complication and readily availability of HPLC, the main objective of present study was to develop a new, precise, accurate, linear, robust, and economical RP-HPLC method for the simultaneous estimation of ertugliflozin and sitagliptin in tablet dosage form. Effective chromatographic separation of Ertugliflozin and Sitagliptin was achieved on Kromasil C18 (5 µm 250 mm X 4.6 mm) and the mobile phase containing Methanol and 0.1% OPA in water isocratic elution mode at a flow rate of 1.0mL/min. with column temperature at 30 °C and the injection volume was 20 µL at column temperature at 30°C. At an isosbestic wavelength of 212 nm, ertugliflozin and sitagliptin were found to have retention times of 5.30 min. and 2.05 min., respectively. The method was proven to be precise (%RSD 2%), accurate (>90%), and specific for the simultaneous measurement of both drugs in tablets. As a result, the suggested method with excellent specificity, accuracy, precision, linearity and robustness as well as economical was useful for the regular quality control analysis of ertugliflozin and sitagliptin tablets.

Application of Principal Component Analysis and DoE-Driven Green Analytical Chemistry Concept to Liquid Chromatographic Method for Estimation of Co-formulated Anti-Hypertensive Drugs.

The fixed-dose combination (FDC) of metoprolol succinate (MTS), cilnidipine (CDN), and telmisartan (TST) is used for the management of hypertension. Numerous reversed phase (RP)-HPLC methods have been reported in the literature for chromatographic analysis of MTS, CDN, and TST. According to the concept of green analytical chemistry (GAC), toxic organic solvents should be avoided or minimized during chromatographic method development for the safety of analysts and the protection of the environment. The reported RP-HPLC methods have been developed using acetonitrile (ACN) or methanol as an organic component of the mobile phase and diluent for sample preparation. These organic solvents are considered toxic solvents as per the International Council for Harmonization (ICH) Q3C (R6) guideline and Pfizer medicinal chemistry solvent selection (PMCSS) guide. We aimed to develop an environment-friendly and economical RP-HPLC-photo-diode array (PDA) method for the analysis of MTS, CDN, and TST using less toxic organic solvents to support the concept of GAC. The method development was carried out by the implementation of chemometrics and design of experiments (DoE) to avoid wastage of organic solvent. Principal component analysis (PCA) was applied as a chemometric tool for the identification of critical method risk variables (MRVs) and method performance attributes (MPAs). The identified critical MRVs and MPAs were further studied by DoE-based response surface modelling for optimization of the method. The chromatographic analysis of MTS, CDN, and TST was carried out using a Shim-pack ODS column as a stationary phase and ethanol as an organic modifier in the mobile phase. The developed method was applied to the assay of FDCs and results were found to be in compliance with the label claim. The greenness profiles of reported and present RP-HPLC methods were evaluated by national environmental method index (NEMI) and analytical greenness (AGREE) methods. The developed method was found to be green, robust, and economical as compared to published methods for the analysis. Development and validation of an RP-HPLC method for simultaneous estimation of MTS, CDN, and TST using safe organic solvents. Implementation of a analytical quality by design (AQbD) approach in method development using PCA and DoE. Application of the method for assay of FDCs of MTS, CDN, and TST.

Method Development and Analysis on High Performance Liquid Chromatograph for the Simultaneous Estimation of Pantoprazole and Itopride in Oral Dosage Form (Tablet)

Attempts were made to develop RP-HPLC method for simultaneous estimation of Itopride HCl & Pantoprazole for the RP - HPLC method, Waters Gradient System with UV Detector and C18 column with 250mm x 4.6 mm i.d and 5μm particle size. After different trial run injections with different solvent mixture ratio and pH of the mobile phase is also alter during one trail condition, From all the traial injection separation of peaks are observed in the mobile phase of the Acetonitrile: 0.05 % OPA (60:40) ration hence this mobile phase ration and pH was used as the mobile phase for the method. The detection wavelength was 252 nm and flow rate was 0.7 ml/min. In the developed method, the retention time of Itopride HCl and Pantoprazole were found to be 4.42 min and 8.59 min. The developed method can be used for the routine quality control analysis of Itopride HCl and Pantoprazole in bulk drug as well as in formulations after complete method Validation.

Application of a Quality by Design-based approach in development and validation of an RP-HPLC method for simultaneous estimation of methotrexate and mangiferin in dual drug-loaded liposomes.

The present study delineates the development of a novel rugged and sensitive stability-indicating risk-based HPLC method for the concurrent estimation of methotrexate and mangiferin in dual drug-loaded nanopharmaceuticals based on an analytical QbD approach. Preliminary screening trials along with systemic risk analysis were performed, endeavouring to explicate the critical method attributes, namely pH, percentage orthophosphoric acid content and percentage methanol content, that influence critical quality attributes. Box-Behnken design was utilized for the optimization of the tailing factor as response for methotrexate and mangiferin in short run time. The chromatographic conditions were optimized by performing 17 experimental runs acquired from Design-Expert software. The chromatographic conditions after the analysis of an optimized zone within the confines of the design space were chosen as mobile phase water-methanol adjusted to pH3.0 with 0.05% orthophosphoric acid (65:35, v/v) and flow rate 1.0ml/min using a C18 analytical column at an isosbestic wavelength of 265 nm. Furthermore, the validation of the optimized method was done in accordance with International Conference on Harmonization guidelines and were reckoned to be in the prescribed limits. The developed RP-HPLC method has a high degree of practical utility for synchronous detection of methotrexate and mangiferin in pharmaceutical nano-dosage forms such as protein-based-nanoparticles, nanocrystals, polymeric nanoparticles and metallic nanoparticles in in vivo and in vitro studies.

RP-HPLC Method Development for Simultaneous Estimation of Oral Iron Chelator Deferiprone and its related Impurity

Purpose: This study aims to develop and validate a RP-HPLC method for simultaneous estimation of deferiprone and its synthesized impurity. Method: Structure of impurity was confirmed by spectral analysis while their purity was confirmed by HPLC analysis. The chromatographic separation was achieved on Agilent Zorbax Bonus-RP (250 x 4.6mm, 5µ) using mobile phase of 20 ACN-80 0.1% TFA. Detection was carried out at 280 nm using PDA detector with flow rate of 1 ml/min and temperature of 30℃. Result: The method was validated as per ICH guidelines. The percent recovery was found to be within limit and %RSD for deferiprone and the impurity was found to be 0.09 and 0.48. The standard curve was linear over the concentration range of 60-140µg/ml and shows correlation coefficient (r2) of 1. The LOD and LOQ values for deferiprone were found to be 0.21µg/ml and 0.62µg/ml respectively. The method was found to be robust even by change in flow rate of ±0.2ml/min. Conclusion: Validation studies demonstrated that the proposed HPLC method is specific, reliable, rapid and reproducible. The developed method can be successfully employed for routine analysis for deferiprone in pharmaceutical dosage form.

Development And Validation of RP-HPLC Method For Simultaneous Estimation of Amlodipine and Valsartan In Its Bulk And Tablet Dosage Form by Using The Quality By Design Approach

Development And Validation of RP-HPLC Method For Simultaneous Estimation of Amlodipine and Valsartan In Its Bulk And Tablet Dosage Form by Using The Quality By Design Approach