scholarly journals DEVELOPMENT AND VALIDATION OF STABILITY INDICATING REVERSE-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF SAQUINAVIR, RITONAVIR, AND AMPRENAVIR

2018 ◽  
Vol 11 (6) ◽  
pp. 390
Author(s):  
Mangamma Kuna ◽  
Gowri Sankar Dannana
Keyword(s):  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Array Detector ◽  
Reverse Phase ◽  
Limit Of Quantification ◽  
Chromatography Method ◽  
Stability Indicating ◽  
Relative Standard ◽  
Liquid Chromatography Method

Objective: The objective of the study was to develop and validate a sensitive, precise, and accurate stability indicating reverse-phase (RP) high-performance liquid chromatography method for the quantification of saquinavir, ritonavir, and amprenavir simultaneously.Methods: The determination of saquinavir, ritonavir, and amprenavir in their mixtures was done using a mobile phase consisted of 0.1M phosphate buffer (pH 3.5) and methanol (70:30, v/v). The method is based on the simultaneous separation of studied drugs in a RP Inertsil ODS C18 (4.6 mm×100 mm, 5 μm) column at ambient temperature. Detection and quantitation were achieved with photodiode array detector set at 260 nm.Results: Saquinavir, ritonavir, and amprenavir showed linearity over a concentration range of 40–200 μg/ml (R2-0.9994), 20–100 μg/ml (R2-0.9992), and 30–150 μg/ml (R2-0.9990), respectively. The limit of quantification was 0.64 μg/ml, 0.57 μg/ml, and 0.53 μg/ml for saquinavir, ritonavir, and amprenavir, respectively. The accuracies for the three drugs were in the range of 99.40–100.53% (saquinavir), 99.45-100.47% (ritonavir), and 100.03–100.53% (amprenavir). The percentage relative standard deviations for the studied drugs were 0.785–0.848% (saquinavir), 0.338–0.499% (ritonavir), and 0.336–0.775% (amprenavir). No peaks were observed at the retention time of saquinavir, ritonavir, and amprenavir in placebo blank, mobile phase blank and stress degraded samples which suggested that the proposed was selective and specific.Conclusion: The method was found to be suitable for the regular analysis of saquinavir, ritonavir, and amprenavir simultaneously in the presence of their stress degradation products.Objective: The objective of the study was to develop and validate a sensitive, precise, and accurate stability indicating reverse-phase (RP) high-performance liquid chromatography method for the quantification of saquinavir, ritonavir, and amprenavir simultaneously. Methods: The determination of saquinavir, ritonavir, and amprenavir in their mixtures was done using a mobile phase consisted of 0.1M phosphate buffer (pH 3.5) and methanol (70:30, v/v). The method is based on the simultaneous separation of studied drugs in a RP Inertsil ODS C18 (4.6 mm×100 mm, 5 μm) column at ambient temperature. Detection and quantitation were achieved with photodiode array detector set at 260 nm. Results: Saquinavir, ritonavir, and amprenavir showed linearity over a concentration range of 40–200 μg/ml (R2-0.9994), 20–100 μg/ml (R2-0.9992), and 30–150 μg/ml (R2-0.9990), respectively. The limit of quantification was 0.64 μg/ml, 0.57 μg/ml, and 0.53 μg/ml for saquinavir, ritonavir, and amprenavir, respectively. The accuracies for the three drugs were in the range of 99.40–100.53% (saquinavir), 99.45-100.47% (ritonavir), and 100.03–100.53% (amprenavir). The percentage relative standard deviations for the studied drugs were 0.785–0.848% (saquinavir), 0.338–0.499% (ritonavir), and 0.336–0.775% (amprenavir). No peaks were observed at the retention time of saquinavir, ritonavir, and amprenavir in placebo blank, mobile phase blank and stress degraded samples which suggested that the proposed was selective and specific. Conclusion: The method was found to be suitable for the regular analysis of saquinavir, ritonavir, and amprenavir simultaneously in the presence of their stress degradation products.

scholarly journals DEVELOPMENT AND VALIDATION OF STABILITY-INDICATING REVERSE-PHASE HIGHPERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF POTENTIAL DEGRADATION PRODUCTS OF REGADENOSON FROM ITS PARENTERAL DOSAGE FORM

2018 ◽  
Vol 11 (8) ◽  
pp. 277
Author(s):  
Murlidhar V Zope ◽  
Rahul M Patel ◽  
Ashwinikumari Patel ◽  
Samir G Patel
Keyword(s):  
Liquid Chromatography ◽  
Degradation Product ◽  
Mobile Phase ◽  
Dosage Form ◽  
Degradation Products ◽  
Reverse Phase ◽  
Chromatography Method ◽  
Stability Indicating ◽  
Liquid Chromatography Method ◽  
Injectable Dosage

Objective: The objective was to develop and validate the stability indicating reverse-phase high-performance liquid chromatography method for the quantification of potential degradation products of regadenoson (REGA) from its injectable dosage form.Methods: YMC-PAK ODS AQ, 150 mm × 4.6 mm, 3 μm composed with hydrophobic high carbon loading and a relatively hydrophilic surface chemically bonded to porous silica particles column was used with the temperature maintained at 40°C. Mobile phase A composed of 0.1% triethylamine buffer having pH 4.5 while mobile phase B is 100 % acetonitrile was used for gradient elution with 1.5 ml/min as a flow rate. The wavelength used for quantification was 245 nm and 20 μl as an injection volume. The suitability of the method has been checked and validated according to the International Council for Harmonization (ICH) guidelines for different parameters, namely, specificity, linearity, accuracy, precision, limit of quantification (LOQ), Limit of detection (LOQ), and robustness studies.Results: The resolution between REGA and its two-degradation product is >8.0 for all pairs of components. The high correlation coefficient (r2>0.990) values are for drug and all potential degradation products from LOQ to 150% of specification limits for impurities calculated based on the maximum daily dose of REGA. LOQ for the drug as well as each degradation product is <0.02% w/w. The % relative standard deviation (RSD) for precision and intermediate precision is in the range of 0.17–0.89, and % RSD for precision at LOQ is 0.86–2.35. The % RSD for robustness study is maximum 2.59.Conclusion: The developed method can quantify the specified and unknown degradation products from 0.1% in the injectable dosage form which indicates that method is sensitive. Method fulfills the ICH criteria for its different validation parameters and demonstrates that the developed analytical method is highly specific, precise, and robust and would have a great value when applied in quality control and stability studies for REGA injection.

scholarly journals Development, Validation, and Routine Application of a High-Performance Liquid Chromatography Method Coupled with a Single Mass Detector for Quantification of Itraconazole, Voriconazole, and Posaconazole in Human Plasma

2010 ◽  
Vol 54 (8) ◽  
pp. 3408-3413 ◽  
Author(s):  
Lorena Baietto ◽  
Antonio D'Avolio ◽  
Giusi Ventimiglia ◽  
Francesco Giuseppe De Rosa ◽  
Marco Siccardi ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Internal Standard ◽  
Limit Of Quantification ◽  
Chromatography Method ◽  
True Value ◽  
Relative Standard ◽  
Mass Detector ◽  
Liquid Chromatography Method

ABSTRACT We have developed and validated a high-performance liquid chromatography method coupled with a mass detector to quantify itraconazole, voriconazole, and posaconazole using quinoxaline as the internal standard. The method involves protein precipitation with acetonitrile. Mean accuracy (percent deviation from the true value) and precision (relative standard deviation percentage) were less than 15%. Mean recovery was more than 80% for all drugs quantified. The lower limit of quantification was 0.031 μg/ml for itraconazole and posaconazole and 0.039 μg/ml for voriconazole. The calibration range tested was from 0.031 to 8 μg/ml for itraconazole and posaconazole and from 0.039 to 10 μg/ml for voriconazole.

Quantification of Pyrazinamide in Human Plasma by Validated High Performance Liquid Chromatography Method

2021 ◽  
Vol 15 (10) ◽  
pp. 2896-2899
Author(s):  
Waleed Arshad ◽  
Naseem Saud Ahmad ◽  
Abdul Muqeet Khan ◽  
Iram Imran ◽  
Qura- Tul-Ain ◽  
...  
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
Mobile Phase ◽  
High Performance ◽  
Ultra Violet ◽  
Chromatography Method ◽  
Ultraviolet Detector ◽  
Relative Standard ◽  
Liquid Chromatography Method

Objective: To be able to accurately determine the quantity of Pyrazinamide (PZA) in different tablet preparations and human plasma using an Ultra violet detector equipped high performance liquid chromatography (HPLC). Study Design: Experimental study Place and Duration of Study: Department of Bioequivalence Studies, University of Veterinary and Animal Sciences Lahore and the Department of Pharmacology, University of Health Sciences, Lahore the from 1st April 2017 to 31st March 2018. Methodology: Two mobile phases were used, the first compromised of disodium hydrogen phosphate buffer having a pH of 6.8 and acetonitrile in the proportion of (95:5) and the second was a combination of aforesaid substances in equivalent proportion (50:50 v/v). The gradient for the first 5 min was exclusively Mobile phase “a” after which 5-6 min Mobile phase “b” was raised from 0 to 100% and was kept at 100% till the completion of the cycle. The flow of mobile phase was kept at 1000 µl/min. Determination of PZA was done using a ultraviolet detector at a wavelength of 238 nm. Amount of sample injected was 40 μl. Procedure was done by using Shizmadu Chromatographic System, Japan equipped with a SIL-20AC HT auto-sampler, SPD-M20A, CTO 20 AC, a LC-20AT VP pump, and CBM 20A controller unit. A C18 column was used as well. Results: Retention time of PZA was 6.1±2%. Precision was 0.46 to 2.20% relative standard deviation for intra assay and for inter assay we obtained 0.29 to 34.45% RSD for all quality control levels. The overall recovery of PZA was 96.75%. Conclusion: High selectivity for PZA was seen and no other spikes from drugs present in FDC regimen were observed at the time when PZA is detected in blank plasma samples Key words: Chromatography, High pressure liquid. Pyrazinamide. Tuberculosis

scholarly journals QUALITY BY DESIGN APPROACH TO STABILITY-INDICATING REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD DEVELOPMENT, OPTIMIZATION, AND VALIDATION FOR THE ESTIMATION OF SIMEPREVIR IN BULK DRUG

2019 ◽  
pp. 93-100
Author(s):  
VANITHA C ◽  
SATYANARAYANA SV ◽  
BHASKAR REDDY K
Keyword(s):  
Liquid Chromatography ◽  
Flow Rate ◽  
High Performance ◽  
Method Development ◽  
Quality By Design ◽  
Polynomial Equation ◽  
Array Detector ◽  
Reverse Phase ◽  
Chromatography Method ◽  
Stability Indicating

Objective: A simple, robust, precise, and selective stability-indicating liquid chromatography (LC) method (reverse-phase high-performance LC) was developed for the estimation of simeprevir through quality by design paradigm. Methods: The chromatographic separation was performed on Water’s 2695(Alliance) equipped with a photodiode array detector at 300 nm. The method was developed on Discovery C18 column (250×4.6, 5 mm) using orthophosphoric acid and acetonitrile (55:45 % v/v) with the flow rate of 1 ml/min at 30°C. The degradation studies of simeprevir were carried out under the stress conditions of hydrolysis (acid, base, and neutral), oxidation, photolytic, and thermal as per the International Conference on Harmonization (ICH) guidelines. The peroxide hydrolysis shows more critical impurities which were well resolved from pure drug with the application of design of experiment and optimized the method. Results: Independent variables (critical analytical attributes) selected for the method optimization were mobile phase ratio, flow rate, and temperature of the column based on the risk assessment. The retention time and resolution were selected as the method response. In response surface method, the central composite design and 23 factorial designs were employed for the optimization of the method. The polynomial equation was derived for the estimation of method response. Conclusion: The method was optimized for better resolution among the drug, and impurity peaks were then validated as per the ICH parameters.

scholarly journals SIMULTANEOUS ESTIMATION OF LOPERAMIDE HYDROCHLORIDE AND TINIDAZOLE IN BULK AND FORMULATIONS BY REVERSE PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY

2016 ◽  
Vol 8 (9) ◽  
pp. 173
Author(s):  
Poornima K. ◽  
Channabasavaraj Kp.
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Limit Of Detection ◽  
Simultaneous Estimation ◽  
Reverse Phase ◽  
Limit Of Quantification ◽  
Chromatography Method ◽  
Ich Guidelines ◽  
Liquid Chromatography Method

<p><strong>Objective: </strong>A new, rapid, selective, precise, accurate and economical, isocratic, reverse phase high-performance liquid chromatography method has been developed for simultaneous estimation of loperamide hydrochloride and tinidazole in bulk and in tablet formulations.</p><p><strong>Methods: </strong>The separation was achieved by using Lithosphere RP C-18, (250 x 4.6 mm, 5 µm) end capped column with a mobile phase containing sodium-1-octane sulfonate buffer: methanol: acetonitrile (40:30:30%v/v/v) pH adjusted to 4.0 (using dilute orthophosphoric acid). The flow rate was 1.0 ml/m and column effluent was monitored at 224 nm. The method was validated as per international conference on chemical harmonization (ICH) guidelines.</p><p><strong>Results</strong>:<strong> </strong>Tinidazole and loperamide hydrochloride were eluted at about 3.1 and 5.4 min respectively, indicating the shorter analysis time. The proposed method was found to be accurate, precise and reproducible. The linearity was established in the concentration range of 10-50 µg/ml. Limit of detection (LOD) and Limit of quantification (LOQ) was found to be 0.001 µg/ml and 0.003 µg/ml for loperamide hydrochloride and 0.01 µg/ml and 0.03 µg/ml for tinidazole.</p><p><strong>Conclusion: </strong>This method can be used for routine analysis of formulations containing any of the above drugs or combinations without any alteration in the chromatographic conditions.</p>

scholarly journals STABILITY-INDICATING REVERSE-PHASE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY METHOD FOR THE SIMULTANEOUS QUANTIFICATION OF APIGENIN AND LUTEOLIN FROM ACHILLEA MILLEFOLIUM LINN

2019 ◽  
pp. 169-172
Author(s):  
GOMATHY SUBRAMANIAN ◽  
S.N.MEYYANATHAN ◽  
GOWRAMMA BYRAN
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Degradation Products ◽  
Forced Degradation ◽  
Reverse Phase ◽  
Chromatography Method ◽  
Stability Indicating ◽  
Degradation Studies ◽  
Liquid Chromatography Method

Objective: A stability-indicating reverse-phase high-performance liquid chromatographic method was developed and validated for the analysis of apigenin and luteolin. The degradation behavior of apigenin and luteolin was investigated under different stress conditions as recommended by the International Conference on Harmonization (ICH). Methods: In the present study, a reversed-phase high-performance liquid chromatography method was developed and the resolution of the plant constituents was successfully achieved using Hibar Lichrospher C8 column with ultraviolet detector at a wavelength of 269 nm. The mobile phase consisted of methanol and 0.5% trifluoroacetic acid (80:20 v/v) at a flow rate of 1.0 ml/min. Both apigenin and luteolin were subjected to various stress degradation studies such as oxidation, acid and alkaline hydrolysis, and photolytic degradation. Results: The proposed method was found to be linear (1–5 μg/ml) with the linear correlation coefficient of R2=0.99. Although the degradation products of stressed conditions were not identified, the methods were able to detect the changes due to stress condition. Conclusion: The method provides good sensitivity and excellent precision and reproducibility. Forced degradation studies on apigenin and luteolin give information about their storage and intrinsic stability conditions considering the advanced pharmaceutical aspects of formulations.

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