Bioanalytical Method Development and Validation of Ranitidine from Plasma Using High Performance Liquid Chromatography (original) (raw)
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International Journal of Pharmacy and Pharmaceutical Sciences, 2017
Objective: This study points to build up and validate a simple methodology to quantify the most used drug sofosbuvir for the treatment of hepatitis C virus (HCV) infection, in human plasma by using atazanavir as an Internal Standard (IS) for preclinical studies and validate as per USFDA guidelines. Methods: Sofosbuvir was isolated from plasma samples by liquid-liquid extraction method using acetonitrile; good chromatographic separation was achieved on Kromasil Column (250 mm ×4.6 mm, 5 µm). The mobile phase consisted of 0.1 % orthophosphoric acid (OPA) buffer pH 2 and acetonitrile in the ratio of (68:32, v/v), respectively. The analysis time was 7 min at a flow rate 1 ml/min. The photodiode array detector (PDA) detection was carried out at 228 nm. The suggested method was validated by performing linearity, system suitability, specificity and sensitivity, accuracy and precision, recovery, ruggedness, stability studies. The method was validated as per USFDA guidelines. Results: The developed method resulted in retention times of sofosbuvir and IS were found out to be 4.7 and 4.2 min respectively. The calibration curves are linear (r2 = 0.999) over the concentration range of 0.050-2.0 µg/ml of plasma analytes concentration. LOQ value was found to be 0.050 µg/ml with precision and accuracy. Within-batch % mean accuracy of the method ranged between 96.00% and 109.09%, and within-batch and total precision, expressed as the coefficient of variation, was 1.40-10.33%. Overall percentage mean recovery of sofosbuvir from spiked plasma was 84.14%. All the validated parameters were found to be within the limit. Conclusion: A simple, accurate, precise, linear, rugged and rapid RP-HPLC method was developed for quantitative estimation of sofosbuvir in human plasma and should be suitable for conducting pharmacokinetics studies and therapeutic drug monitoring.
Journal of Advanced Scientific Research
A Reverse Phase High Performance Liquid Chromatographic (RP-HPLC) method was developed and validated for theestimation of Sofosbuvir which is a new antiviral drug. The RP-HPLC was succeeded on a Phenomenex Luna® LC C18column (150×4.6mm, 5μm). The mobile phase was effective according to the polarity of studied drug. The mobile phasewas consisting of Acetonitrile: Methanol: Water in the ratio of 50:30:20 v/v/v using at flow rate of 1ml/min. withinjection volume of 20 μL was selected for this present work. Detection was made by using UV detector at 260nm.Retention time was found to be 2.1min. The developed method was validated according to the ICH guidelines. Thecalibration curve was linear for Sofosbuvir in the concentration range of 10-50 μg/ml was good. The developed methodwas validated for Linearity, Precision, Accuracy and Robustness of Sofosbuvir drug and was accurate, precise and reliablefor the analysis of Sofosbuvir in formulation. The Relative Standard Deviation for all the p...
Objective:Simple, rapid and reproducible reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet derivative spectrophotometric (UVDS) methods for the simultaneous determination of daclatasvir (DAC) and sofosbuvir (SOF) in pure and in pharmaceutical dosage forms have been developed and validated. Methods:The chromatographic separation of DAC and SOF was achieved using Agilent Zorbax SB C 18 (4.6 x 250 mm, 5 µm) column at temperature of 40 ° C. The mobile phase used was 9 mM dipotassium hydrogen orthophosphate buffer (pH 4±0.1): acetonitrile (60:40, v/v). The flow rate was maintained at 1 mL/min with UV detection at 265 nm. Results:The calculated resolution was 4.56 (> 2), which ensures complete separation. The tailing factor was 1.13 and 1.40 (≤ 2) for DAC and SOF, respectively. Intermediate precision value was ≤ 2% indicate acceptable ruggedness. Conclusion:The two methods were validated according to the International Conference on Harmonization (ICH) guidelines in terms of linearity, precision, accuracy, selectivity, specificity, detection limit, quantification limit, robustness and ruggedness.
Estimation and Validation of Sofosbuvir in Bulk and Tablet Dosage Form by RP-HPLC
International Journal of Pharmacy, 2016
A simple, sensitive, precise, and accurate isocratic reverse phase high pressure liquid chromatographic method has been developed and validated for the estimation of sofosbuvir in bulk and tablet dosage form. To optimize, a column Phenomenex prodigy ODS-3V (150 mm x 4.6 mm, 5 µm), mobile phase mixture of methanol and (0.1%) tri-fluro acetic acid as buffer having pH of 3.2 in the ratio of (30:70 v/v) found to be an efficient system for elution of drug with good peak shape as well as retention time 2.990 min., flow rate 1.0 ml/min. at UV wavelength of 260nm. Quantitative linearity was obeyed in the concentration range of 100 to 600 µg/ml, the regression equations of concentration over their peak areas were found to be Y = 18864x + 58306 R² = 0.996, where Y is the peak area and X is the concentration of drug. The number of theoretical plates obtained was 2604.352 which indicate the efficient performance of the column. The limit of detection was 0.01 µg/ml and limit of quantification was 0.03 µg/ml, which indicates the sensitivity of the method the high percentage recovery indicates that the proposed method is highly accurate. No interfering peaks were found in the chromatogram indicating that excipients used in tablet formulation did not interfere with the estimation of the drug by the proposed RP-HPLC method.
A simple, specific and accurate reverse phase ultra-performance liquid chromatographic method was developed for the simultaneousdetermination Sofosbuvir and Velpatasvirin pharmaceutical dosage form. The column used was Kromosil C18(150mm x 4.6 mm, 5µm)inisocratic mode, with mobile phase containing phosphate buffer andacetonitrile(75:25v/v). The flow rate was 1.0ml/ min and effluents weremonitored at 260 nm. The retention times of Sofosbuvir and Velpatasvirwere found to be 2.404min and 2.986 min, respectively.The linearityfor Sofosbuvir and Velpatasvirwere in the range of 35-210µg/ml and 8-48 µg/ml respectively. The recoveries of Sofosbuvir andVelpatasvirwere found to be 99.64% and 99.25%, respectively. The proposed method was validated and successfully applied to the estimationofSofosbuvirandVelpatasvirincombinedtabletdosageforms.
Asian Journal of Pharmaceutical and Clinical Research, 2016
ABSTRACTObjective: The objective of the present work is to develop a simple, efficient, and reproducible spectrophotometric method for the quantitativeestimation of hepatitis-C drugs - Daclatasvir and Sofosbuvir in its active pharmaceutical ingredient (API) form.Methods: The developed ultraviolet spectrophotometric method for the quantitative estimation of hepatitis-C drugs - Daclatasvir and Sofosbuvir isbased on measurement of absorption at a wavelength maximum (λmax) of 317 and 261 nm using methanol as solvent.Results: The method was validated in terms of specificity, precision, linearity, accuracy, and robustness as per the ICH guidelines. The method wasfound to be linear in the range of 50-150% for Daclatasvir and in the range of 43-143% for Sofosbuvir. The percentage recovery values were in therange of 99.4-100.6% for Daclatasvir and in the range of 99.7-100.6% for Sofosbuvir at different concentration levels. Relative standard deviation forprecision and intermediate precision ...
International Journal of Pharmaceutical Sciences, 2024
Background: To assess the concentration of sofosbuvir (CAS-1190307-88-0) in pharmaceutical formulations as well as in its pure form, a unique green analytical chemistry approach was developed. ATR-FTIR technique was used to quantitatively evaluate sofosbuvir, a multipurpose antiviral drug, in both tablet and bulk forms. Objective: Sofosbuvir in pharmaceutical goods may be quickly detected by the development and confirmation of an accurate, sensitive, and non-invasive attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy approach. Methods: Sofosbuvir is analysed using a method that looks for the distinctive skeletal band for CO -C stretching in the 1170-1000 cm-1 spectral region. These procedures, which have linear ranges for SBR of 1.0 to 7.0 mg g-1, were fully verified in compliance with ICH recommendations. Results: The correlation value was 0.9993, and the linearity ranged from 1.0 to 7.0 milligrammes. The results indicated that the limits of quantification (LOQ) and detection (LOD) were 0.142 ± 0.09 mg and 0.079 ± 0.02 mg, respectively. Conclusions: Green chemistry guided the development of the analytical approach, and it was shown that the outcomes were exactly in line with destructive methods. Highlights: It was shown that the results produced were in perfect accord with destructive methods, and the analytical strategy was designed in light of green chemistry.
BMC Chemistry
In the present study two different RSLC columns, Acclaim RSLC 120 C18, 5.0 µm, 4.6 × 150 mm (column A) and Acclaim RSLC 120 C18, 2.2 µm, 2.1 × 100 mm (Column B) were utilized for the analysis of velpatasvir (VPS) in presence of sofosbuvir (SFV), where due to the encountered fluorescent properties of VPS fluorescent detection at 405 nm after excitation at 340 nm (Method 1) was used for its detection where the non-fluorescent SFV did not interfere. The same columns were further utilized for the simultaneous determination of SFV and VPS either in bulk form or in their combined tablet, where UV-spectrophotometric detection at 260 nm was selected for the simultaneous analysis of both drugs (Method 2). A mobile phase consisting of NaH 2 PO 4 , pH 2.5 (with phosphoric acid) and acetonitrile in a ratio of 60:40 v/v was used for both methods. The mobile phase was pumped at a flow rate of 1.0 mL/min when using column, A and 0.5 mL/min when using column B. The methods showed good linearity over the concentration ranges of 1.0-5.0 and 2.5-10.0 ng/mL for VPS when utilizing Method 1 A and B respectively. Where the linearity concentration range was from 30.0-150.0 to 120-600.0 ng/mL for VPS and SFV respectively when applying Method 2. Both methods 1 and 2 were performed by utilizing the two analytical columns. The different chromatographic parameters as retention time, resolution, number of theoretical plates (N), capacity factor, tailing factor and selectivity were carefully optimized. The results show that comparing the performance of the two utilized columns revealed that shorter column (2.1 mm × 100 mm) with small particle packing was superior to the longer column (4.6 × 150 mm) for the analysis of the studied drugs allowing a reduction of the analysis time by 70% without any detrimental effect on performance. This prompts the decrease of the investigation costs by saving money on organic solvents and expanding the overall number of analyses per day.
The present research work describes a simple, accurate, precise and effective UV-Vis Spectroscopic and RP-HPLC method for simultaneous estimation of Sofosbuvir and Ledipasvir. Spectrophotometric methods for simultaneous estimation are developed & validated using Q-Absorbance ratio and dual wavelength method. In Q-Absorbance ratio method Sofosbuvir and Ledipasvir showed an iso-absorptive point at 241nm, the second wavelength used was 260nm, which was λmax of Sofosbuvir. In dual wavelength method Sofosbuvir was measured at 252 nm and 310.70 while Ledipasvir was measured at 332 nm and 310.70 nm. A reverse phase high performance chromatographic method was developed for simultaneous estimation of Sofosbuvir and Ledipasvir in their combined dosage. The separation was achieved by BDS Hypersil C18 column (150X4.6mm, 5µm) column, and ACN : 0.1% TFA in the proportion of 32:68 %v/v as mobile phase, at a flow rate of 1 ml/min. Detection was carried out at 245 nm.
International journal of health sciences
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. An accurate, precise and reproducible UV spectrophotometer method was developed for the simultaneous quantitative determination of Sofosbuvir and Ledipasvir in tablet dosage forms. Methods: Younglin (S. K.) gradient system UV detector and C18 column with 250 mm x 4.6 mm i. d. and 5μm particle size Acetonitrile: OPA water (80: 20v/v) pH 2.5 was used as the mobile phase for the method. The detection wavelength was 283 nm and flow rate was 0.9 ml/min. Results: In the developed method, the retention time of Sofosbuvir and Ledipasvir were found to be 6.366 min and 8.616 min. The developed method was validated ...