LC-MS/MS method for oseltamivir and metabolite in plasma.
Journal name: World Journal of Pharmaceutical Research
Original article title: A selective and sensitive liquid chromatographic/ tandem mass spectrometric method for simultaneous estimation of oseltamivir and its metabolite oseltamivir carboxylic acid in human plasma for bioavailability or bioequivalence studies
The WJPR includes peer-reviewed publications such as scientific research papers, reports, review articles, company news, thesis reports and case studies in areas of Biology, Pharmaceutical industries and Chemical technology while incorporating ancient fields of knowledge such combining Ayurveda with scientific data.
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Shankrappa Janiwarad, Kashif Ul Haq, Dinesh Choudhary
World Journal of Pharmaceutical Research:
(An ISO 9001:2015 Certified International Journal)
Full text available for: A selective and sensitive liquid chromatographic/ tandem mass spectrometric method for simultaneous estimation of oseltamivir and its metabolite oseltamivir carboxylic acid in human plasma for bioavailability or bioequivalence studies
Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research
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Summary of article contents:
Introduction
Oseltamivir, a neuraminidase inhibitor effective against influenza A and B, is administered as the prodrug oseltamivir phosphate. Upon oral intake, it is rapidly absorbed and metabolized by hepatic esterases primarily into its active form, oseltamivir carboxylate (OCA). This study focuses on developing and validating a sensitive liquid chromatographic/tandem mass spectrometric (LC-MS/MS) method for the simultaneous quantification of oseltamivir and its metabolite OCA in human plasma, which is essential for conducting bioavailability and bioequivalence studies. The method adheres to US FDA guidelines and aims to address previous deficiencies observed in similar analytical methods.
Method Development and Validation
The developed method utilized a solid phase extraction approach with Oasis® HLB cartridges for the extraction of 0.5 mL plasma samples. Chromatography was performed on a Hydrosphere C18 column, and a mobile phase of acetonitrile and 0.1% formic acid was implemented. The quantification was conducted on a triple quadruple API-3200 LC/MS/MS system with a positive electrospray ionization source. The method’s selectivity, sensitivity, precision, recovery, and stability were meticulously validated. Linear calibration curves for oseltamivir and OCA were established, demonstrating high coefficients of correlation, indicating the method's reliability for quantitative analysis.
Sensitivity and Selectivity
The sensitivity of the analytical method was showcased through low limits of quantification (LLOQ), demonstrating acceptable precision and accuracy. The selectivity was confirmed by analyzing six different blank plasma samples, which exhibited no interfering peaks at the analytes' retention times, ensuring the method's reliability in discerning oseltamivir and OCA from other endogenous substances present in human plasma. Furthermore, the matrix effect analysis revealed that the presence of plasma components did not significantly affect the analyte ionization, thus supporting the method's validity for real sample applications.
Stability and Reliability
Extensive stability studies indicated that both oseltamivir and OCA remained stable under various storage conditions, including freeze-thaw cycles and room temperature for extended periods. The method demonstrated its robustness, consistently yielding reproducible results across multiple validation batches and even post-reanalysis, which highlights its applicability in pharmacokinetic studies and routine clinical analysis. A rigorous approach to extending batch verification indicated the method's capability to handle a high volume of samples while maintaining accuracy, thereby streamlining the analytical processes in clinical settings.
Conclusion
The validated LC-MS/MS method for the simultaneous determination of oseltamivir and its metabolite, OCA, stands as a reliable analytical solution for pharmacokinetic studies. This method not only complies with regulatory requirements but also addresses limitations observed in previously reported methods, such as specificity and recovery rates. Its simplicity and efficiency in analysis mark a significant advancement in bioanalytical methods, making it a valuable tool for drug monitoring and clinical research focused on oseltamivir treatment efficacy.
FAQ section (important questions/answers):
What was the main objective of the study?
The main objective was to develop and validate a method for simultaneously estimating Oseltamivir and its metabolite in human plasma for bioavailability or bioequivalence studies.
What extraction method was used in the study?
A single-step solid phase extraction method using Oasis® HLB cartridges was employed to extract Oseltamivir and its metabolite from human plasma samples.
How were the analytes quantified in plasma samples?
Quantification was performed using a triple quadruple API-3200 LC/MS/MS with positive electrospray ionization in multiple reaction-monitoring mode.
What were the validation parameters for the method?
The method was validated for selectivity, sensitivity, accuracy, precision, recovery, stability, and ruggedness, according to US FDA guidelines.
What were the linear ranges for Oseltamivir and its metabolite?
The linear range was 2.08 ng/mL to 241.12 ng/mL for Oseltamivir and 10.8 ng/mL to 1251.8 ng/mL for Oseltamivir carboxylic acid.
What stability conditions were tested for the analytes?
Stability assessments included freeze-thaw cycles, bench-top stability, post-extraction stability, and long-term stability at -70°C.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “LC-MS/MS method for oseltamivir and metabolite in plasma.”. This list explains important keywords that occur in this article and links it to the glossary for a better understanding of that concept in the context of Ayurveda and other topics.
1) Table:
In scientific research, a 'Table' is an organized display of data, often used to summarize and present findings in a clear format. In this method validation context, tables are used to illustrate results such as recovery rates and precision metrics, making complex data easily accessible for analysis and comparison.
2) Quality:
'Quality' refers to the standard of the results obtained from the method validation process. High quality in analytical methods ensures that the estimates of Oseltamivir and its metabolite are accurate, reproducible, and reliable, which is crucial for bioavailability and bioequivalence studies to meet regulatory standards.
3) Water:
'Water' is a critical solvent in analytical chemistry, often used in buffer solutions and sample preparations. In this study, HPLC-grade water ensures that the extraction and chromatography processes are free from contaminants, contributing to the integrity of the mass spectrometric analysis of Oseltamivir and its metabolite.
4) Study (Studying):
'Study' indicates a systematic investigation, often designed to test a hypothesis or evaluate a theory. In this context, the study focuses on validating a liquid chromatographic/tandem mass spectrometric method to assess the pharmacokinetics of Oseltamivir, pivotal for ensuring effective therapeutic applications.
5) Post:
'Post' typically refers to activities occurring after a significant event or process. In this text, it may relate to 'post-processing' which involves analysis or stability assessments following sample preparation, ensuring that the samples maintain their integrity and the results are consistent over time.
6) Drug:
'Drug' is a term referring to substances used for medical purposes. In this study, Oseltamivir is a drug used to treat influenza, and understanding its pharmacokinetics is essential for evaluating its efficacy and safety in clinical settings, especially during bioavailability studies.
7) Purity:
'Purity' pertains to the concentration of the active ingredient without contamination from other substances. In the method validation process, ensuring the purity of standards like Oseltamivir and OCA is crucial for accurate quantification and reliable analytical results, directly impacting therapeutic assessments.
8) Blood:
'Blood' is a vital biological matrix used for pharmacokinetic studies. In this context, the analysis of Oseltamivir and its metabolite in human plasma (derived from blood) provides insights into drug metabolism and distribution, fundamental for determining how effectively the drug works in patients.
9) India:
'India' is mentioned in the context of sourcing chemical reagents. The reference to Varada Biotech (P) Ltd from Mumbai signifies the global nature of pharmaceutical research, where collaborations and sourcing of high-quality standards for analytical methods often involve international partnerships and local expertise.
10) Noise:
'Noise' in analytical chemistry refers to unwanted signals that can interfere with the detection of analytes. In method validation, maintaining a high signal-to-noise ratio is critical for the selective detection of Oseltamivir and its metabolite, ensuring that the results are accurate and devoid of background interference.
11) Viru (Vīṟu):
'Viru' likely refers to 'virus,' specifically considering Oseltamivir's role as a neuraminidase inhibitor against influenza viruses. The study's focus on this drug aligns with its therapeutic targets in viral infections, highlighting its relevance in public health and epidemic management strategies.
12) Discussion:
'Discussion' typically involves interpreting the results and the implications of the findings in scientific literature. In this context, the discussion would be important for analyzing the method's performance, comparing it with other methods, and affirming its applicability in bioanalytical settings.
13) Medicine:
'Medicine' pertains to the science and practice of diagnosing, treating, and preventing disease. This study contributes to the field of medicine by validating a method crucial for assessing the therapeutic efficacy of Oseltamivir, guiding clinical decisions and medication management in influenza cases.
14) Activity:
'Activity' in pharmacology refers to the capacity of a drug to produce a biological effect. This study assesses the pharmacological activity of Oseltamivir through detailed bioanalytical methods, helping to characterize its therapeutic potential and effectiveness in real-world medical scenarios.
15) Relative:
'Relative' is often used in scientific measurements to compare values against a standard. In bioanalytical contexts, relative measurements of drug concentrations inform pharmacokinetic profiles, helping to understand dosage requirements and therapeutic effects in patient populations.
16) Varada (Varadā, Vāraḍa, Vara-da):
'Varada' refers to Varada Biotech (P) Ltd, a supplier of reference standards mentioned in this study. Their involvement underlines the importance of sourcing high-purity substances for analytical validation, which is crucial for ensuring the reliability of the results obtained in the pharmacokinetic evaluations.
17) Liquor:
'Liquor' typically refers to a liquid substance. In this analytical context, 'liquor' may pertain to 'liquor ammonia' used as a reagent. Its role in sample preparation or analytical processes is essential, highlighting the variety of chemicals involved in chromatographic and mass spectrometric analysis.
18) Food:
'Food' is included to possibly reference food-grade chemical compliance for reagents. In pharmaceutical studies, ensuring that reagents meet safety and quality standards, akin to food safety regulations, reinforces the credibility and safety of the research, particularly when translating findings to human health.
19) Line:
'Line' in this context may refer to the calibration line used in the method validation process. Establishing a reliable calibration line ensures accurate quantification of analytes, such as Oseltamivir and its metabolite, facilitating consistent and reproducible results in pharmacokinetic studies.
Other Science Concepts:
Discover the significance of concepts within the article: ‘LC-MS/MS method for oseltamivir and metabolite in plasma.’. Further sources in the context of Science might help you critically compare this page with similair documents:
Best practice, Clinical studies, High performance liquid chromatography, Drug stability, Mass spectrometry, Retention time, Liquid chromatography-tandem mass spectrometry, Internal standard, Multiple reaction monitoring, Calibration curve, Method validation, Linear regression analysis, Chromatographic condition, Bioavailability studies, Pharmacokinetic studies, Reference Standard, Signal to noise ratio, Goodness of fit.