RP-HPLC method for stability, estimation of lopinavir and ritonavir.

| Posted in: Science

Journal name: World Journal of Pharmaceutical Research
Original article title: Stability indicating method development and validation for simultaneous estimation of lopinavir and ritonavir by using rp-hplc
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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Original source:

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Author:

P. S. Raghu


World Journal of Pharmaceutical Research:

(An ISO 9001:2015 Certified International Journal)

Full text available for: Stability indicating method development and validation for simultaneous estimation of lopinavir and ritonavir by using rp-hplc

Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research

Doi: 10.20959/wjpr20183-11106


Download the PDF file of the original publication


Summary of article contents:

Introduction

Lopinavir and Ritonavir are widely recognized antiretroviral agents used in combination therapy for treating HIV/AIDS. This study, conducted by P.S. Raghu at the University College of Pharmaceutical Sciences, focuses on developing and validating a rapid and sensitive reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of these two drugs in pharmaceutical formulations. The methodological optimization involves various chromatographic conditions to ensure accuracy, precision, and practicality for quality control testing in the pharmaceutical industry.

Method Development and Optimization

The developed RP-HPLC method employs an Inertsil C18 analytical column with a mobile phase composed of orthophosphoric acid buffer and acetonitrile in a specific ratio. An analytical setup featuring a flow rate of 1.0 mL/min and a system maintained at 30°C was utilized. The method's optimal detection wavelength for both drugs was determined to be 310 nm, with retention times recorded at 2.214 minutes for Lopinavir and 3.163 minutes for Ritonavir. This optimization resulted in a method that is not only efficient but also suitable for regular quality control testing.

Validation of the Method

A significant aspect of the research was the method's validation, which included assessing parameters such as accuracy, precision, and linearity. The accuracy was demonstrated at varied concentrations with recovery rates of 99.64% for Lopinavir and 99.56% for Ritonavir, reinforcing the reliability of the results. Additionally, the method's precision was confirmed through repeated tests, yielding low relative standard deviation (RSD) values, indicating high reproducibility in results.

Analytical Performance and Robustness

The analytical performance of the RP-HPLC method was thoroughly evaluated, highlighting its linearity across a range of concentrations. Furthermore, robustness experiments demonstrated that small intentional changes in chromatographic conditions did not significantly impact the method's performance, underscoring its stability and reliability. The limits of detection (LOD) and quantification (LOQ) were established, with Lopinavir at 0.20 ppm and 0.61 ppm, and Ritonavir at 0.08 ppm and 0.24 ppm, respectively, affirming the method’s sensitivity.

Conclusion

The newly developed RP-HPLC method for the simultaneous estimation of Lopinavir and Ritonavir is effective, accurate, and suitable for routine quality control analyses in pharmaceutical settings. Its validation and robustness highlight its potential for widespread application in the pharmaceutical industry. The method's simplicity and economic nature further enhance its feasibility for analyzing commercial products, offering a reliable option for ensuring the quality of antiretroviral medications.

FAQ section (important questions/answers):

What it the purpose of the developed RP-HPLC method?

The developed RP-HPLC method aims to accurately and precisely estimate Lopinavir and Ritonavir simultaneously in pharmaceutical dosage forms, providing a simple and economical approach for quality control testing.

What are the optimal conditions for the method?

The optimal conditions include a mobile phase of ortho phosphoric acid buffer and acetonitrile at a 48:52 ratio, pH 6.0, a flow rate of 1.0 mL/min, and a wavelength of 310 nm for detection.

How reliable is the method's accuracy and precision?

The method demonstrated an accuracy of 99.64% for Lopinavir and 99.56% for Ritonavir, with high precision indicated by %RSD values of 0.6 and 0.3, respectively.

What were the retention times for Lopinavir and Ritonavir?

The retention times found were 2.214 minutes for Lopinavir and 3.163 minutes for Ritonavir, making them easily identifiable during analysis.

What are the limits of detection and quantification?

The limit of detection (LOD) for Lopinavir is 0.20 ppm and for Ritonavir is 0.08 ppm. The limits of quantification (LOQ) are 0.61 ppm and 0.24 ppm, respectively.

What materials were used in the method development?

Reference standards of Lopinavir and Ritonavir were obtained from Tenna labs, along with HPLC grade solvents from Merck India and analytical grade chemicals from SD Fine Chemicals.

Glossary definitions and references:

Scientific and Ayurvedic Glossary list for “RP-HPLC method for stability, estimation of lopinavir and ritonavir.”. 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:
A table is a structured arrangement of data, typically consisting of rows and columns. In the context of research, tables present numerical information, statistics, and experimental results concisely and clearly, allowing for easy comparison and analysis of different parameters such as recovery percentages, precision, and limits of detection or quantification.

2) Raghu:
Raghu is the surname of the lead author of the study discussed in the article. P.S. Raghu is a researcher associated with the University College of Pharmaceutical Sciences. His expertise and leadership are crucial for conducting the experiments, analyzing data, and interpreting results related to the simultaneous estimation of Lopinavir and Ritonavir.

3) Drug:
The term 'drugs' refers to multiple substances used for medicinal purposes. In the research context, it emphasizes the need for reliable methods to evaluate the effectiveness, dosage, and safety of pharmaceuticals, particularly combinations of drugs like Lopinavir and Ritonavir that are essential in managing viral infections.

4) Performance:
Performance refers to the effectiveness and efficiency of a method or system in producing desired outcomes. In this study, performance metrics such as precision, accuracy, and recovery rates are critical in validating the developed RP-HPLC method, ensuring it meets the standards required for pharmaceutical quality control.

5) Purity:
Purity indicates the degree to which a substance is free from impurities or contaminants. In pharmaceutical analysis, ensuring high purity for drugs like Lopinavir and Ritonavir is essential as it affects efficacy and safety. The study assesses the purity of the samples using HPLC, emphasizing reliability in measurements.

6) Krishnadevaraya:
Sri Krishnadevaraya University in Andhra Pradesh, India, is the academic institution where the study was conducted. Named after a historical king, the university plays a crucial role in higher education and research, providing an environment for scientific investigations like the simultaneous estimation of drugs for quality assurance.

7) Study (Studying):
A study is a systematic investigation aimed at discovering or confirming knowledge. In this context, the study develops and validates an RP-HPLC method for analyzing Lopinavir and Ritonavir, showcasing rigorous research practices to produce reliable and reproducible results vital for pharmaceutical analysis and validation.

8) Science (Scientific):
Science is the systematic enterprise that builds and organizes knowledge in the form of testable explanations and predictions. In the article, scientific principles are applied to develop analytical methods for pharmaceuticals, emphasizing the role of scientific inquiry in improving public health through better drug quality and efficacy.

9) Quality:
Quality refers to the standard of excellence or grade of a product or process. In the pharmaceutical context, it encompasses the safety, efficacy, and reliability of drugs. Ensuring quality is paramount for regulatory compliance and patient safety, which is the focus of validation in the study's methodology.

10) India:
India is a country where the study was conducted, hosting various institutions that contribute to pharmaceutical research. The significance of this study lies in advancing healthcare by developing accurate analytical methods in a country that increasingly focuses on improving access to medications and enhancing public health standards.

11) Water:
Water is a crucial solvent in the pharmaceutical industry, often used in the preparation of solutions for analytical methods. The document mentions its use in HPLC procedures, emphasizing the need for high-quality water for accurate results, as impurities in water can significantly affect analytical outcomes.

12) Measurement:
Measurement refers to the determination of the size, amount, or degree of something. In this study, accurate measurement of drug concentrations using RP-HPLC is critical. Reliable measurements ensure that the pharmaceutical formulations meet required standards and thus maintain therapeutic effectiveness and patient safety.

13) Discussion:
Discussion refers to the section of a research paper where results are interpreted and implications are explored. This part of the article allows authors to connect findings with existing knowledge, analyze the significance of the results, and propose future research directions, thereby enriching the scientific discourse.

14) Relative:
Relative signifies a comparison between two or more entities based on their relationships or ratios. In the context of the study, relative parameters may include the purity percentages, recovery rates, and performance metrics of the analytical method for Lopinavir and Ritonavir, allowing for assessment against standards.

15) Babu:
Babu is a common South Asian surname, here referring to Sudhakar Babu, who is acknowledged for his support in the research study. Recognition of contributors highlights the collaborative nature of scientific research, emphasizing mentorship and institutional backing in successfully conducting and publishing research endeavors.

Other Science Concepts:

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Discover the significance of concepts within the article: ‘RP-HPLC method for stability, estimation of lopinavir and ritonavir.’. Further sources in the context of Science might help you critically compare this page with similair documents:

High performance liquid chromatography, Retention time, Mobile phase, Limit of detection, Limit of quantification, Chromatographic parameters.

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