RP-HPLC Method For Estimation Research Articles

Development and Validation of RP HPLC Method for Estimation of Deferiprone and Its Related Impurity in Pharmaceutical Dosage Form

Development and Validation of RP HPLC Method for Estimation of Deferiprone and Its Related Impurity in Pharmaceutical Dosage Form

Development and Validation of RP HPLC Method for Estimation of Deferiprone and its Related Impurityin Pharmaceutical Dosage Form

Aim of this study is to develop a new, precise, sensitive, simple, efficient, selective, and accurate high-performance liquid chromatographic method for the separation and determination of Deferiprone and its impurity in the capsule dosage form. A wide-range of literature survey disclosed no method for estimation said as the above. The chromatographic separation was achieved on Agilent Zorbax Bonus-RP (250 x 4.6mm, 5µ) with a mobile phase of Methanol: 0.1% O-Phosphoric acid (10:90, % v/v) combination in 1000ml of Methanol: Water (50: 50, % v/v) using a diluent. The flow rate of 1mL/min and UV detection at 280nm use as wavelength. The developed method was validated as reported by ICH guidelines. The linearity of the calibration curve for deferiprone and its process-related impurity in the concentration range of 4.0-6.0μg/ml. There exists a good correlation between peak area and analyte concentration. The retention time for deferiprone was discovered to be 2.29 min and its impurity was 8.65min. Deferiprone's relative standard deviation value is 0.45 and its process-related impurity is 0.17. All the results tell that the proposed method was highly sensitive, simple, precise, accurate, and fast. A large number of samples can be analyzed in a shorter time due to shorter retention times, so it can be successfully applied for routine analysis of Deferiprone and related impurity (maltol) in pharmaceutical dosage forms.

Development and Validation of RP-HPLC Method for estimation of Secnidazole in API and Pharmaceutical Dosage Form

Objective of the present work is to develop and validate a simple, cost effective, sensitive and fast HPLC method for the analysis of Secnidazole. A Merck-Hitachi HPLC system with Peerless Basic C18 (50mm x 4.6mm x 3μm) column is employed for the analysis using buffer: methanol (80:20, v/v) as mobile phase. Signal from Secnidazole is detected at 310nm by UV Spectrophotometer. The proposed method is fully validated and found to be linear over a workable drug concentration, accurate, precise and robust. This fast and inexpensive method is suitable for research laboratories as well as for quality control analysis in pharmaceutical industries.

Development and validation of stability-indicating RP-HPLC method for estimation of dalfampridine in bulk drug and tablet dosage form

BackgroundIn the current study, a simple, improved, precise, rapid, and accurate reverse phase liquid chromatographic method was produced for the estimation of dalfampridine in bulk and tablet dosage form which is a potassium channel blocker used for the treatment of multiple sclerosis (MS). The separation of dalfampridine was achieved isocratically on a C18 column (250 × 4.6 mm, 5 μm) using (0.1% v/v) buffer pH 3.0 ± 0.05 adjusted with diluted orthophosphoric acid (OPA) and acetonitrile (ACN) in the ratio of 60:40% (v/v) as a mobile phase, at a flow rate of 0.5 mL/min, and column temperature of 40 °C. HPLC grade methanol as diluents was used. Five microliters of the standard solution of the drug was injected, and the eluted analytes were detected at 262 nm.ResultsDalfampridine was eluted at 4.5 min with a run time of 10 min. Linearity in the method was measured in the concentration range of 25–75 ppm with a correlation coefficient of 0.999. Limit of detection and limit of quantitation were found to be 0.711 μg/mL and 2.154 μg/mL, respectively. Dalfampridine was subjected for forced degradation stability study in conditions of thermal, acid, alkali, and oxidation and photo-degradation condition. The degradants were well resolved from the dalfampridine main peak. Validation of the developed method is carried as per USFDA and ICH guidelines.ConclusionThe results of the analysis prove that the method is simple, improved, precise, accurate, and rapid for estimating the content of dalfampridine in bulk drug and tablet dosage form and can be applied for routine analysis.

Development and Validation of RP-HPLC Method for Estimation of Methotrexate Drug Intercalated in Mg-Al Layered Double Hydroxide Nanoparticles

In the present study, pristine magnesium aluminium layered double hydroxide (Mg-Al LDH, sample A) nanoparticles were synthesized by simple co-precipitation technique followed by intercalation of methotrexate drug (MTX), their detailed physicochemical characterization and development of RP-HPLC method for estimation of MTX intercalated in Mg-Al LDH nanoparticles (sample B). The PXRD analysis showed the basal spacing of 003 plane corresponding to sample A and sample B to be 8.63 Å and 11.39 Å, respectively. Particle size distributions were measured to be 56-75 nm (D50=66.15 nm) in case of sample A and 75-100 nm (D50=87.41 nm) for sample B. Simple, accurate RP-HPLC method was developed for estimation of the MTX drug from sample B, on Agilent HPLC 1200 system with UV detector on L1 column. 0.02 M potassium dihydrogen phosphate solution (pH 3.0) and acetonitrile (85:15 v/v) was used as mobile phase in an isocratic elution mode at a flow rate of 1.0 mL/min at 35°C with a load of 10 μL at 307 nm. The retention time for methotrexate was at 4.44 min. The method was validated as per ICH Q2 (R1) guidelines. The method was found to be linear over a concentration range of 10-100 μg/mL.

Development and Validation of RP-HPLC Method for Estimation of Ondansetron HCL in Bulk and Pharmaceutical Dosage Form

Development and Validation of RP-HPLC Method for Estimation of Ondansetron HCL in Bulk and Pharmaceutical Dosage Form

QbD-Based Development and Validation of an Efficient RPHPLC Method for Estimation of Abiraterone Acetate in Bulk, Tablet, and In-House-Developed Nano-Formulation

The present study focused on the development of a simple, reliable, cost-effective, precise, and stable quality by design (QbD) RP-HPLC method for estimation of Abiraterone Acetate in bulk, pharmaceutical dosage form, and in-house-developed nano-formulation. The essential analytical target profile (ATP), and critical analytical attributes (CAAs) were described. Plackett-Burman Design and Box-Behnken Design were used for screening and optimization of critical method parameters (CMPs). The chromatographic method with reverse-phase isocratic mode, C-18 column, 69: 31 (% v/v) acetonitrile, and ammonium acetate buffer (pH 3.5) as a mobile phase with a flow rate of 0.75 ml/min and 235 nm detection wavelength was developed. The method was validated as per International Conference on Harmonization guideline and found to be linear between 10 to 180 μg/ml with R2 0.997, detection limit of 3.64 μg/ml, and quantitation limit of 11.05 μg/ml. Precision was demonstrated using a relative standard deviation of 1.55 % and 1.59 % for inter and intraday studies respectively. The developed method was also used for the analysis of ABT in bulk, tablet dosage form, and in-house-developed nanosponges prepared using different cross-linking agents, devoid of any interference of formulation excipients. The method effectively established the utility of QbD for optimizing chromatographic conditions used for the estimation of Abiraterone acetate.

Chromatographic method for estimation of vitamin E from dried blood spot sample

BackgroundWe developed a selective bioanalytical RP-HPLC method for estimation of vitamin E from dried blood spot (DBS) sample, a potential technique which can be used for population-based epidemiological studies. Vitamin E was extracted from DBS by using liquid-liquid extraction technique with methanol (100% v/v) as reconstituting solvent for the residue. Alpha tocopheryl acetate was used as internal standard. Samples were analyzed directly on HPLC with C18 (250 × 4.6 mm × 5 μm) Phenomenex column. The mobile phase used was methanol to water (99:1% v/v) at a flow rate of 1.4 mL/min. The detector wavelength used was 292 nm.ResultsThe retention time observed for vitamin E and internal standard was 10.225 ± 0.00075 min and 13.580 ± 0.00075 min respectively. The vitamin E calibration curve was found to be linear over the range of 0.625 to 60 μg/mL. The limit of quantification for vitamin E was found to be 0.1 μg/mL. Accuracy of the developed method was found to be 103.179%, 101.625%, and 100.174% with percentage of coefficient of variation of 0.0161, 0.0215, and 0.2790 for HQC, MQC, and LQC samples respectively which were within USFDA acceptance limit of ± 15 to ± 20%.The intraday and interday precision expressed as coefficient of variation were 0.0191–0.0841% and 0.0074–0.0252% respectively.ConclusionsThe method represents a simple, rapid, specific, accurate, and precise method for estimation of vitamin E in human blood using DBS technique. The developed method can be further evaluated with respect to effect of matrix variability before it can be used in clinical setting.

Method Development and Validation of Irbesartan by RP-HPLC Method

This study includes development of RP-HPLC method for estimation of Irbesartan. The developed method was validated as per ICH guidelines in terms of specificity, accuracy, linearity, LOD, LOQ, ruggedness and robustness. The assay experiment showed that the contents of Irbesartan estimated in the tablet dosage were free from interference of excipients, which indicate that developed method was specific. Recovery of standard added drugs was found to be 97.2–99.9% for Irbesartan indicating that the proposed method was accurate. After performing analysis by different analyst; it was found that the RP-HPLC method for the determination of Irbesartan % RSD of ruggedness for standard 0.0425% and tablet 0.0142% was found to be rugged. % RSD of robustness was well within the limits for standard 0.36% and tablet 1.87% that the proposed method was robust. The LOD was 0.074 μg/ml for standard drug of Irbesartan. For standard drug of Irbesartan the LOQ were found to be 0.24 μg/ml This demonstrated that the developed RP-HPLC method was simple, linear, precise, accurate, robust and rugged, could be conveniently adopted for the routine quality control analysis of Irbesartan from it's pharmaceutical dosage form and bulk drug.

RP-HPLC Method for estimation of Alogliptin and Glibenclamide in Synthetic mixture

The objective of the current study was to develop simple, accurate, precise and rapid RP-HPLC method with subsequently validate as per ICH guidelines for the determination of Alogliptin benzoate and Glibenclamide hydrochloride using mobile phase [mixture of Acetonitrile: Phosphate buffer pH-3.6 and in the ratio of 65:35.The proposed method involves the measurement of Retention time at selected analytical wavelength. 240.0 nm. The retention time of Alogliptin benzoate (ALO) and Glibenclamide was found to be 5.055 and 2.838 respectively. The linearity of the proposed method was investigated in the range of 1-5 µg/ml (r = 0.9998) for ALO and 10-50µg/ml (r = 0.9999) for MET respectively. The method was statistically validated for linearity, accuracy and precision. Both inter-day and intra-day variation was found to be showing less % RSD (Relative Standard Deviation) value indicating high grade of precision of the method.

Development and Validation of RP-HPLC Method for Estimation of Etoposide in Liposomes

Development and Validation of RP-HPLC Method for Estimation of Etoposide in Liposomes

Comparison of UV-spectrophotometric and RP-HPLC methods for estimation of deflazacort in solid dosage form

Comparison of UV-spectrophotometric and RP-HPLC methods for estimation of deflazacort in solid dosage form

Quantitative bioanalytical and analytical methods for estimation of ivabradine hydrochloride in pure and pharmaceutical dosage form

Introduction: New analytical and bioanalytical methods were developed for the estimation of Ivabradine hydrochloride in bulk and pharmaceutical dosage form by UV spectrophotometry and high-performance liquid chromatography (HPLC) technique. Objective: The primary objective of the study is to develop a new RP-HPLC method for estimation of Ivabradine Hydrochloride in pure and formulation and to develop a bioanalytical method for analysis of Ivabradine Hydrochloride in biological samples. The methods were validated as per International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use and USFDA guidelines respectively. Methods: In Reversed-phase (RP)-HPLC method developed telmisartan is the internal standard used. After liquid–liquid extraction, the analyte and the internal standard were chromatographed on Waters 125A (10 µ, 300 × 3.9 mm) C18 column using 20-µL injection volume with a run time of 15 min. An isocratic mobile phase consisting of acetonitrile and ammonium acetate buffer pH 7.8 (60:40% vol/vol) is used to separate drug and internal standard. In Spectrophotometric bioanalytical method developed for the estimation of ivabradine hydrochloride in pure and pharmaceutical dosage form. The solvent system used is absolute methanol and detected at the wavelength of 287 nm. The solvent system used is absolute methanol and detected at the wavelength of 287 nm. Results: The Analytical method is validated according to ICH guidelines over the range of 2–16 μg/ mL, showing accuracy, precision, selectivity, and robustness. For Bioanalytical method the linearity is established in the range of 500–3500 ng/mL with the regression coefficient of r2 = 0.9994. The validated spectrophotometric method is used successfully to study ivabradine hydrochloride in rat plasma and also quantitative determination in marketed tablets. Conclusion: The proposed methods were successfully applied for the quantitation of ivabradine hydrochloride in pharmaceutical dosage form with good recovery and reproducibility.

A Validated RP-HPLC Method for Estimation of Related Compounds in Hydroxy Naproxen

A simple, linear gradient liquid chromatographic method (RP-HPLC) is proposed for the simultaneous estimation of related compounds in hydroxy naproxen samples. Chromatographic separation was achieved on Zorbax SB C8 (150 × 4.6) mm, 3.5 μm particle size RRLC short column and eluent A used as 0.1% v/v trifluoroacetic acid in water and eluent B used as acetonitrile using Agilent RRLC (UHPLC) system. The mobile phase flow rate was 1.0 mL/min and the eluted compounds were monitored at 235 nm for related substance method and assay method. The excellent resolution was obtained between hydroxy naproxen and its related compounds, which were eluted within 25 min. The performance of the method was validated with respect to ICH guidelines for specificity, limit of detection, limit of quantification, linearity, accuracy, precision and robustness. The correlation coefficient(r) was > 0.995 for both the methods from linearity data and percentage of recovery is 98.0 to 102.0 for assay method and 80.0 to 120.0% for related substance method. Sensitivity of the method was found be less than 0.5 μg/mL. Peak homogeneity data for naproxen in the chromatograms from the selectivity solution obtained by use of the photodiode array detector demonstrated the specificity of the method for analysis of hydroxy naproxen in presence of the related compounds

Development and Validation of RP HPLC Method for Estimation of Vortioxetine in Bulk and Pharmaceutical Dosage Form

Development and Validation of RP HPLC Method for Estimation of Vortioxetine in Bulk and Pharmaceutical Dosage Form

RP-HPLC Stability Indicating Method Development and Validation of Pseudoephedrine Sulfate and Related Organic Impurities in Tablet Dosage Forms, Robustness by QbD Approach

Pseudoephedrine is used for the temporary relief of stuffy nose and sinus pain/pressure caused by infection such as the common cold and flu. Here we were reporting a simple and precise validated stability indicating RP-HPLC method for estimation of Pseudoephedrine Sulfate and its related compounds in solid oral dosage forms. The stability indicating nature of current method has been demonstrated by a forced degradation study. All process related impurities and degradation products from forced degradation samples were well resolved from each other. The chromatographic separation of impurities was achieved with a Hypersil phenyl column (25 cm x 4.6 mm i.d., particle size 5 μ) using buffer and acetonitrile in the ratio of 95:5 v/v respectively as the mobile phase. The buffer used in mobile phase contained 10 mM potassium dihydrogen phosphate and its pH was adjusted to 3.0 ±0.05 with diluted orthophosphoric acid. The flow rate of HPLC was 1.0 mL min−1 and the wavelength of UV detector at 210 nm. The method was shown a linear response over range of 0.125 μg mL−1 to 0.750 μg mL−1 PES and impurities. The correlation coefficient was more than 0.999. Performed the robustness by using design experiments.

Development and Validation of a Stability Indicating RP-HPLC Method for Estimation of Etizolam in Tablet Dosage Form

A simple, precise, and sensitive stability indicating liquid chromatographic method was developed and validated for Etizolam in tablets. An isocratic RP-HPLC method was developed for Etizolam on a C18 Agilent (150 x 4.6 mm x 3.5µm) column using a mobile phase consisting of acetonitrile and triethylamine solution, pH 3.2 (50:50, v/v) at a flow rate of 1.0 mL/min. The chromatographic peaks for Etizolam were eluted at 3.21 ± 0.24 min and the detection was carried out at 254 nm. The method obeyed linearity in the range of 0.5 - 100 µg/mL (r2=0.999). The method gave repeated results as observed from the % RSD (< 2.0) indicating the precision. The mean recovery obtained in the present method was found to be 98.58 ± 0.32%. The drug was subjected to a variety of stress environments and was highly sensitive to acid, base and oxidation conditions. The method was validated as per ICH guidelines in terms of specificity, accuracy, precision, LOD, LOQ and robustness. This method can be used for quantification of Etizolam in tablet dosage form without any interference from excipients.

Development of Stability Indicating RP-HPLC Method for Estimation of Clemastine Fumarate and its Pharmaceutical Dosage Form

A simple, fast and precise stability indicating RP-HPLC method was developed for quantification of Clemastine Fumarate in pure and pharmaceutical dosage form. The quantification was carried out using LiChrospher® 100 RP-C8 endcapped column (5μm, 150 × 4.6mm) and mobile phase comprised of methanol and water (0.05% triethyamine) in proportion of ratio 90: 10 and degassed under ultrasonication. The flow rate was 0.8ml/min and effluent was monitored at 220nm. The retention time of Clemastine Fumarate was found to be 7.49±0.039 min and peak was separated. The method was validated in terms of linearity, precision, accuracy, specificity, robustness, limit of detection and limit of quantitation in accordance with ICH guideline. Linearity of Clemastine Fumarate was in the range 15–50μg/mL. The percentage recovery of Clemastine Fumarate were 98.46 to 99.99% from tablet formulation. The stability-indicating capability was established by forced degradation experiments. The proposed method suitable for determination of Clemastine Fumarate in pharmaceutical dosage form.

A Validated RP-HPLC Method for Estimation of Telmisartan and Metoprolol in its Bulk Form

A Validated RP-HPLC Method for Estimation of Telmisartan and Metoprolol in its Bulk Form

Simple Extraction Cum RP-HPLC Method for Estimation of Nanotized Quercetin in Serum and Tissues of Mice

In recent years, several studies have focused on antioxidant, anti-inflammatory, and anti-cancer activities of quercetin (3,3′,4′,5, -pentahydroxyflavone). The nanotization of quercetin was shown to enhance its therapeutic efficacy due to smaller particle size. In the present study, an additional step was added to simple extraction cum RP-HPLC method for the quantification of nanotized quercetin (nQ) in biological samples to understand the pharmacokinetics and biodistribution of nQ following intravenous administration. The proposed method involves extraction of nQ from blood serum and tissues of mice with 2N HCl in comparison to well-known DMSO:MeOH mix method. The HCl extraction was found to be 2 – 3 times more efficient than DMSO:MeOH mix method. Results showed that the amount of nQ at various time intervals in the serum and tissues was 2 – 3 fold greater for HCl extraction than for DMSO:MeOH mix method, suggesting that HCl extraction must take into account nQ bound with protein. The reversed-phase HPLC was used for nQ detection, which showed the nQ retention time of 3.2 min. The limit of detection of nQ in blood serum was found to be 0.1 μg/mL. The proposed method was also validated in terms of linearity, precision, and accuracy.