Development Of RP-HPLC Method Research Articles

A new analytical RP-HPLC method development and validation for the estimation of Nilutamide in bulk form and marketed pharmaceutical dosage form

A rapid and precise reverse phase high performance liquid chromatographic method has been developed for the validated of Nilutamide, in its pure form as well as in tablet dosage form. Chromatography was carried out on a Symmetry C18 (4.6 x 150 mm, 5 µm) column using a mixture of Methanol and water (45: 55 % v/v) as the mobile phase at a flow rate of 0.8 ml/min, the detection was carried out at 260 nm. The retention time of the Nilutamide was 2.379 ± 0.02 min respectively. The method produce linear responses in the concentration range of 24-120 mg/ml of Nilutamide. The method precision for the determination of assay was below 2.0 % RSD. The method is useful in the quality control of bulk and pharmaceutical formulations. The method was validated for accuracy, precision, linearity, robustness, ruggedness and LOD & LOQ of standard solution. The developed RP-HPLC method was found to be accurate, precise, linear, and robust and was successful applied to a pharmaceutical tablet formulation for qualitative estimation of Nilutamide in Bulk form and Marketed Pharmaceutical Dosage forms.

Box-Behnken Design employed stability demonstrating RP-HPLC method development of sorafenib tosylate for rapid and sensitive quantification

Sorafenib tosylate (SFN) is a tyrosine kinase inhibitor used clinically to treat liver, kidney, and thyroid carcinoma. This research aims to accurately quantify SFN using a quality-by-design (QbD) approach with RP-HPLC. Chromatographic settings were optimized using a Box-Behnken design, measuring responses such as retention time, tailing factor, theoretical plate, and peak area. At the same time, independent variables were flow rate, mobile phase composition, and wavelength. A C18 column (4.6 × 250 mm, 5 µm) served as the stationary phase, with a mobile phase of methanol and 0.1% o-phosphoric acid in a 41:59 v/v ratio. SFN detection occurred at 267 nm in isocratic mode at a flow rate of 1.1 mL min−1. Method validation followed International Council for Harmonization guidelines, yielding a coefficient of determination (R2) of 0.9998, indicating linearity in the 5-25 µg mL−1 range. Results showed detection (LOD) and quantification (LOQ) limits of 0.04 and 0.12 µg mL−1, respectively. Additionally, the method demonstrated precision, accuracy, and robustness consistent with ICH criteria. Overall, this simple, accurate, rapid, and robust RP-HPLC method is suitable for routine SFN analysis in various formulations.

Development of RP-HPLC Method of Tizanidine HCL with Some Validation Parameter.

This study compiles the information for the development of analytical methods for estimation of the Tizanidine HCl that will be helpful for further research work on this drug and its impurity. The present Literature survey provides information about the Analytical methods like UV,TLC,RP-HPLC,HPTLC,UHPLC and other methods have been reported for Tizanidine HCl drug individually and along with other drugs. The analysis of published data revealed that, there was only UV spectroscopic method (calibration curve metod) is reported for estimation of Tizanidine HCl fixed dose combination. Estimation of Tizanidine HCl by superlative RP-HPLC method i.e. Mobile phase- Acetonitrile: phosphate buffer (pH: 7.5) (50:50%v/v), Column C18 (250mm*4mm*5µm), Flow rate- 1.0 ml/min, Wavelength: 318 nm. Optimized HPLC condition was validated by assessing validation parameters and it meet the acceptance criteria set by ICH. It was showed method was linear and precise. The validated RP-HPLC-PDA method can be used for routine analysis of Tizanidine HCl in tablet.

RP-HPLC Method Development and Validation for the Estimation of Trelagliptin in Pure and Pharmaceutical Dosage Form

RP-HPLC Method Development and Validation for the Estimation of Trelagliptin in Pure and Pharmaceutical Dosage Form

Development and Validation of a Robust RP-HPLC Method for the Simultaneous Analysis of Exemestane and Thymoquinone and Its Application to Lipid-Based Nanoformulations.

The present study describes the development and validation of a simple and rapid HPLC method for the simultaneous quantification of exemestane and thymoquinone. The separation of both compounds was performed on a 5 μ C-18 column utilizing phase A as water/methanol (45:5 v/v) and phase B as acetonitrile (50 v/v) (total ratio of A/B = 40:60 v/v) in isocratic elution mode as the mobile phase at a flow rate of 0.8 mL/min. Further, the Box-Behnken design was used for optimizing the analytical method. The proposed method was validated for various parameters, and all parameters were found to be within an acceptable range. The simultaneous detection of both drugs was monitored at 243 nm with a retention time of 5.73 and 6.93 min, respectively. Moreover, the forced degradation studies were conducted under various stress conditions, and the relevance of the validated RP-HPLC method was further explored for the estimation of drugs from lipid-based nanoformulation. Taken together, the study construed the development of an efficient and robust method that could be used for the quantification of these agents in various in vitro as well as in vivo models.

Exploration of AQbD Approach for Development of RP-HPLC Method of Nebivolol

Exploration of AQbD Approach for Development of RP-HPLC Method of Nebivolol

Stability Indicating RP-HPLC Method Development and Validation for Imeglimin HCL in Pharmaceutical Dosage form

Background: Imeglimin HCL is a medication used in pharmaceutical products. To ensure its quality and effectiveness, it's crucial to have a reliable method to measure its concentration in these products. High-Performance Liquid Chromatography (HPLC) is a common technique for this purpose. Objective: This study aimed to develop and validate a precise and accurate stability indicating HPLC method for measuring Imeglimin HCL in pharmaceutical formulations. Methods: The method optimization process included selecting a suitable chromatographic column and determining the optimal mobile phase composition. A Credchrom C18 column (250mm x 4.6 mm x 5µm) was chosen, and a mobile phase consisting of Phosphate Buffer and acetonitrile (80:20, v/v) was identified as optimal. The selected conditions provided satisfactory resolution and a retention time of 2.5 minutes for Imeglimin HCL, with detection achieved at a wavelength of 241nm. Validation parameters, including linearity, precision, accuracy, limit of detection (LOD), and limit of quantification (LOQ), were thoroughly evaluated. Precision was assessed through %RSD values for repeatability and intra-day and inter-day precision. Accuracy was determined by % recovery, with values ranging from 98.00% to 102%. Subsequently, force degradation studies were conducted to further assess the method's stability. Results: The developed RP-HPLC method exhibited excellent linearity (R^2 close to 1) and sensitivity, with LOD and LOQ values of 0.577722 μg/ml and 1.750673 μg/ml, respectively. Precision, as indicated by %RSD values, ranged from 0.994733 for repeatability to 0.988377–0.7480963 for intra-day precision and 0.988377–10.9883477 for inter-day precision. Accuracy, reflected by % recovery, was within acceptable limits (98.62–100.34%). These results, along with successful force degradation studies, confirmed the method's reliability and stability. Conclusions: In conclusion, the developed RP-HPLC method offers a sensitive, precise, and accurate means for the estimation of Imeglimin HCL in pharmaceutical formulations. Its robustness and stability make it suitable for routine analysis in pharmaceutical laboratories, ensuring the quality and efficacy of Imeglimin-containing products.

RP-HPLC Method Development and Validation for Simultaneous Estimation of Erlotinib and Ramucirumab in bulk and Pharmaceutical Dosage form and its stability studies

RP-HPLC Method Development and Validation for Simultaneous Estimation of Erlotinib and Ramucirumab in bulk and Pharmaceutical Dosage form and its stability studies

RP-HPLC Method Development and Validation of Choline Salicylate and Lignocaine HCl in Mouth Ulcer Gel

RP-HPLC Method Development and Validation of Choline Salicylate and Lignocaine HCl in Mouth Ulcer Gel

A New RP-HPLC Method development and validation for Etodolac and Thiocolchicoside

Abstract The aim of this work is to develop and validate a simple, sensitive, rapid and accurate and less time consuming validated RP-HPLC method for Etodolac and Thiocolchicoside in pharmaceutical dosage form. The method was developed and validated for various parameters as per ICH guidelines. The results obtained were within the acceptance criteria. The proposed method was applied for the determination of Etodolac and Thiocolchicoside in marketed formulation. The assay results confirm with the label claim of formulation.. Hence, the proposed method was found to be satisfactory and could be used for the routine analysis of Etodolac and Thiocolchicoside in combined tablet dosage forms. The retention time were found to be was 2.777 and 2.380 min for Etodolac and Thiocolchicoside respectively. The quantitative estimation gave a satisfactory result for Etodolac (99.66 % w/w) and Thiocolchicoside (99.3% w/w) respectively. The regression values over its peak areas were found to y = 17537x + 361.5 and y = 17769x + 1505 respectively and correlation coefficient found to be about 0.9999 and 0.9998 for Etodolac and Thiocolchicoside respectively. The percentage recovery for Etodolac and Thiocolchicoside were found to be 99.99% and 101.2 % respectively

Stability Indicating RP-HPLC Method Development and Validation for Simultaneous Estimation of Metformin Hydrochloride and Remogliflozin Etabonate in Bulk and Tablet Formulation

A simple efficient stability-indicating validated RP-HPLC method has been developed for the simultaneous determination of Metformin hydrochloride (MET) and Remogliflozin etabonate (REM) in bulk and was applied to marketed formulations. The mobile phase used for detection was Phosphate Buffer (pH 4.0): Acetonitrile (60:40%v/v). Drug peaks were detected by a UV detector at 226nm. The linearity of the developed method was found to be 20-60µg/mL for Metformin hydrochloride and 4-12μg/mL for Remogliflozin etabonate. The Limit of Detection (LOD.) and Limit of Quantitation (LOQ.) were found to be 0.085µg/mL and 0.258µg/mL for MET and 0.010µg/mL and 0.030µg/mL for REM respectively. Hydrolysis by HCl, NaOH, hydrogen peroxide, UV light and temperature were performed on a formulation which proves that the proposed method was specific and will be suitable for routine quality control analysis.

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).

Rp-HPLC method development and validation of finerenone in bulk drug and its formulations

I have done research and achieved the method development & validation of highly accurate, sensitive, precise, rapid gradient system RP-HPLC method of finerenone in both bulk drug & pharmaceutical fixed dosage forms. The separation was achieved on (Agilent) C18 column (4.6 mm*250 mm, 5µm) having this configuration with particle size 5µm & using mobile Methanol: 10 Mm Citric Acid with ratio (38:62) having flow rate 1.0 ml/min. Detection of finerenone was carried out at 258nm. The total chromatographic analysis time per sample was about 15min and retention time of overall analysis is 6.9 min. The response exhibited a linear relationship with concentration in range 98-102 mcg/ml for finerenone correlation coefficient is 0.999 and %RSD is 0.126. The method was validated for accuracy, precision, specificity, linearity & sensitivity. Validation studies demonstrated that the method is simple, specific, rapid, reliable & reproducible.

Development and validation of a QbD-based RP-HPLC method for vericiguat quantification

Aim: An RP-HPLC method for Vericiguat using the QbD approach was developed and validated by ICH guidelines. Method: The ICH (Q2R1) guidelines have been followed in the development and validation of an RP-HPLC technique by considering several validation parameters like linearity, precision, LOD, LOQ, and accuracy. The study was performed on Agilent Tech using the C18 column (4.6x250 mm; 5 µm) and Chemstation 10.1 software with statistical data analysis, and the detector used was UV (DAD). Results: The mobile phase used for separation was Methanol: 0.1% OPA in the ratio of (76:24) at room temperature, the flow rate was 0.8ml/min, and the wavelength was 331nm. The results indicated that the quantification limit was 0.7209 µg/ml, and the detection limit was 0.2379 µg/ml. Conclusion: The validation studies confirmed that the developed method is fast, accurate, precise, cost-effective, selective, and useful for routine analysis of vericiguat in tablet dosage forms.

Simultaneous Estimation of Naltrexone Hydrochloride and Zonisamide in Synthetic Mixture by RP-HPLC Method

A simple, rapid, precise, and reproducible RP-HPLC method development and validation done for simultaneous estimation of Naltrexone Hydrochloride and Zonisamide in synthetic mixture. The isocratic method was developed usinɡ a Water Symmetry C18 (250 mm X 4.6 mm, 5 µm) column. Mobile phase composition was Phosphate Buffer: Methanol (75:25 %V/V) (Buffer-1.36 ɡ of KH2PO4 in 1000 ml adjust pH 3.0 ± 0.05 with OPA.), flow rate 1 ml/min and detection carried out at 230 nm at column temperature 30ºC. The injection volume was 10µl with run time of 10 minutes. Parameters such as linearity, precision, accuracy, recovery, specificity and robustness are performed as per ICH ɡuidelines Q2(R1). Retention time of Naltrexone Hydrochloride and Zonisamide were found to be 3.92 min and 8.15 min respectively. The developed method found linear in the ranɡe of 1-10 µɡ/ml for Naltrexone Hydrochloride and 2.84-142 µɡ/ml for Zonisamide (R2 = 0.9999 for both druɡs). The % Recovery were found to be 99.39 %-100.70 % and 98.91 %-100.59 % for Naltrexone Hydrochloride and Zonisamide respectively. The RSD for both druɡ were less than 2 for precision, specificity and robustness.

RP-HPLC Method Development and Validation of Moxifloxacin Hydrochloride & Ketorolac Tromethamine in Bulk and its Pharmaceutical Formulation

RP-HPLC Method Development and Validation of Moxifloxacin Hydrochloride & Ketorolac Tromethamine in Bulk and its Pharmaceutical Formulation

Development and Validation of RP-HPLC Method using UV detection for Simultaneous quantification of Amlodipine besylate and Nebivolol hydrochloride in Fixed-dose combination tablets

Calcium channel blockers and beta blockers are the class of choice of drugs in treatment of hypertension and cardio vascular diseases. Such a fixed dose combination is viable to deal with the patients for whom monotherapy is not sufficient. A simple, responsive and productive high performance liquid chromatography method was designed and approved for simultaneous quantification of Amlodipine Besylate (AMB) and Nebivolol Hydrochloride (NBL) in tablet dosage form. In the developed method mobile phase comprised of a mixture of aqueous buffer (pH 6) and acetonitrile (55:45 v/v, respectively) at a flow rate of 1.5 ml/min. Chromatographic separation was achieved using Reverse-phase Kromasil 60-5-CN column, 250 × 4.6 mm (particle size 5 μm). Method was discovered specific as no obstruction of excipients peaks at the retention time of tablets was observed and demonstrated linearity in concentration range of 30.4-70.8 mcg/ml for AMB and 29.8-69.4 mcg/ml for NBL with R2> 0.999. Accuracy values for AMB and NBL was found to be 98.21-99.49% and 97.91-99.09%, respectively. Inter-day and intra-day precision was carried out and % RSD was found to be less than 2%. Robustness was performed to evaluate the results with slight variations in method and % RSD found to be less than 2%. Solution stability studies were conducted in normal conditions. Force degradation study was carried out using Photo-diode Array (PDA) detector and revealed that acid degradation in AMB and NBL was 7.12% and 1.88%, alkaline degradation was 9.61% and 0.05% and oxidative degradation was 8.50% and 1.59%, respectively.

Development of a fast RP-HPLC method for the separation of nifurtimox and its forced degradation products through a design of experiment approach

The study depicts the successful development and validation of a simple and fast reverse-phase high-performance liquid chromatography (RP-HPLC) method for the separation of nifurtimox (NFX) and its degradation products. NFX was exposed to different degradation conditions and a RP-HPLC method reported in the literature was used to evaluate different peaks that could interfere with the one from the pure NFX. NaOH generated a degradation product, with a peak that tends to overlap with NFX. Therefore, the NFX exposed to this condition was used during screening and optimization of the RP-HPLC method using a Design of Experiment (DoE) approach. A factorial design was applied to screen different factors, followed by a central composite design for process optimization with those factors that significantly affected the evaluated responses during the screening step. The optimized method was further validated according to International Conference of Harmonization guidelines. Methanol proportion and gradient time to reach it, alongside with mobile phase flow rate, were the factors that had a statistically significant effect on resolution, retention time, and tailing factor in the chromatographic peak. The optimized chromatographic parameters were: 41.6 % of methanol, a gradient time of 2 min, and a flow rate of 0.62 mL/min. Under these chromatographic conditions, the retention time of NFX was 5.3 min, while for the nearest degradation products was 4.5 min and 6.3 min. The resolution of the NFX peak was 5.14 and the tailing was 1.13. The DoE approach allowed the development of an optimized RP-HPLC method capable of successfully separating NFX and its forced degradation products.

Development and Validation of RP-HPLC Method for Routine Analysis of Osimertinib

Osimertinib is a newly approved treatment line for the non-small cell lung carcinoma actively caused by mutation in EGFR and T790M. The present study was targeted to originate a validated method for the qualification as well as quantification of osimertinib through RP-HPLC in bulk and its pharmaceutical formulation. The mobile phase, 65% triethylamine buffer (pH 2.5) and 35% acetonitrile (v/v) was run through a prontosil column (C18, 4.6×150 mm i.d., 3.5 μm particle size) at a flow rate of 0.7 ml/min. A PDA plus detector was used to scan the absorbance over 210 to 360 nm. Validation study performed following the ICH guidelines Q2 (R1) was found the developed method to be specific, precise, accurate, linear over the range of 64–96 μg/ml with R2 > 0.999, robust and rugged. The % recovery of osimertinib at different levels ranged between 99.32% and 100.37% from tablet formulation (Osicent). The method holds promise for analysis of osimertinib in bulk, pharmaceutical formulations and for further research. Bangladesh Pharmaceutical Journal 27(1): 85-91, 2024 (January)

Development and Validation of RP-HPLC Method for Analysis of Counterfeit Clopidogrel

Development and Validation of RP-HPLC Method for Analysis of Counterfeit Clopidogrel