Method development and validation of metformin by HPLC.

| Posted in: Science

Journal name: World Journal of Pharmaceutical Research
Original article title: Method development and validation of metformin by using reverse- phase high performance liquid chromatography
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:

Krishna Mohan Chinnala, B. Parijatha, D. Santhoshi Priya and Alapati Sahithi


World Journal of Pharmaceutical Research:

(An ISO 9001:2015 Certified International Journal)

Full text available for: Method development and validation of metformin by using reverse- phase high performance liquid chromatography

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

Doi: 10.20959/wjpr20178-9069


Download the PDF file of the original publication


Summary of article contents:

Introduction

Metformin, an oral anti-diabetic drug belonging to the biguanides class, plays a crucial role in managing type 2 diabetes, particularly in patients who are obese. Its efficacy has led to its availability in over 60 countries worldwide. The development of a reliable method for analyzing Metformin is vital, as current chromatographic approaches can be complex and time-consuming. This article presents a study focusing on the development and validation of a sensitive and efficient method for Metformin quantification using Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) in conjunction with Ultraviolet (UV) spectrophotometry.

Method Development and Optimization

The method employs HPLC with a HITACHI L2130 system, utilizing a C18 Develosil ODS column and an isocratic mobile phase consisting of acetonitrile and phosphate buffer in a 30:70 ratio. During the initial phases of method development, parameters such as flow rate, wavelength, and injection volume were optimized. The selected detection wavelength was 225 nm, with a flow rate of 1.0 ml/min. In the developed method, Metformin was effectively quantified after employing specific dilutions and sample preparations, leading to well-resolved chromatograms demonstrating a retention time of approximately 2.80 minutes.

Method Validation and Performance

The validation of the developed method showcased high linearity, with a coefficient of determination (r²) of 0.998 over the range of 0-100 µg/ml, establishing its reliability for quantitative analysis. Recovery studies demonstrated accuracy, with recovery rates averaging around 100 percent for different concentrations of Metformin spiked into pre-analyzed samples. Precision values for both intra-day and inter-day assays showed minimal variability, thus affirming the robustness and reliability of the method under varied conditions.

Robustness and Stability Testing

Robustness studies assessed the method's resilience to minor alterations in operational parameters—flow rate, temperature, and wavelength variations. Results indicated that the % relative standard deviations were remarkably low, affirming the method's stability. Additionally, assays conducted on commercial formulations of Metformin confirmed the method's practicality, yielding consistent results aligned with stated drug content. System suitability tests, including resolution and theoretical plate counts, further reinforced the method's efficacy.

Conclusion

The study successfully developed and validated a sensitive, selective RP-HPLC method for analyzing Metformin in pharmaceutical formulations. The method's accuracy, precision, and robustness make it ideal for routine quality control and stability testing. As such, it represents a valuable analytical tool for ensuring the quality and efficacy of Metformin products in various formulations available in the market.

FAQ section (important questions/answers):

What is Metformin and its classification in pharmaceuticals?

Metformin is an oral anti-diabetic drug classified as a biguanide, primarily used for treating type 2 diabetes. It functions by reducing glucose production in the liver and enhancing insulin sensitivity.

What methods were used for Metformin analysis in this study?

The study developed and validated a method using reverse-phase high-performance liquid chromatography (RP-HPLC) and ultraviolet (UV) spectrophotometry for the analysis of Metformin, ensuring it is effective for routine analysis.

What were the optimal chromatographic conditions for Metformin analysis?

Optimal chromatographic conditions included a C18 Develosil ODS column, a mobile phase of acetonitrile and phosphate buffer (30:70), a flow rate of 1.0 ml/min, and a wavelength of 225 nm.

How was the accuracy of the method verified?

Accuracy was determined through recovery studies, adding known amounts of pure Metformin to a pre-analyzed sample. The recovery percentage was calculated to confirm the method’s reliability.

What was the correlation coefficient for Metformin's calibration curve?

The calibration curve for Metformin demonstrated excellent linearity with a correlation coefficient (r²) of 0.998 over the range of 0-100 µg/ml.

What key results affirmed the method's precision in the study?

The study reported low relative standard deviations (% RSD) for repeatability and intra-day/inter-day precision, confirming that the developed method is precise for Metformin analysis.

Glossary definitions and references:

Scientific and Ayurvedic Glossary list for “Method development and validation of metformin by HPLC.”. 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:
Table refers to a systematic arrangement of data, often organized in rows and columns. In the context of this research article, tables are used to present instrumental data, chemicals, results, and other relevant information clearly and concisely, facilitating easier understanding and comparison of data findings related to Metformin analysis.

2) Drug:
Drug denotes any substance used in the diagnosis, treatment, or prevention of diseases. In this paper, Metformin is the central drug under study, recognized for its role as an oral hypoglycemic agent in managing type 2 diabetes, impacting patient health significantly and requiring rigorous analysis for safety and efficacy.

3) Water:
Water serves as a universal solvent essential in biochemical reactions and laboratory experiments. It is mentioned in the context of solubility studies of Metformin, highlighting its fractions in various solvents, which is crucial to understanding how the drug behaves in a biological environment and during preparations in pharmaceutical analyses.

4) Purity:
Purity indicates the degree to which a substance is free from contaminants and impurities. In pharmaceutical contexts, assessing drug purity is imperative, as the presence of impurities may affect the drug's efficacy and safety; the study emphasizes the importance of developing methods for verifying the purity of Metformin preparations.

5) Study (Studying):
Studying is a systematic process of learning and analysis. In this document, it reflects the ongoing research efforts to explore Metformin's efficacy and safety through various analytical methods, reinforcing the importance of rigorous study protocols to ensure the reliability and validity of outcomes in pharmaceutical science.

6) Performance:
Performance refers to how well a method or system operates to produce desired outcomes. In the text, performance is evaluated in relation to the HPLC method, which is assessed for its sensitivity, accuracy, and reproducibility in analyzing Metformin, ultimately determining its practicality for routine pharmaceutical use and regulatory compliance.

7) Narasimha (Narasingha, Nara-simha):
Narasimha likely refers to a personal or institutional name associated with the educational context of the research. This name indicates the affiliation of the authors with Nalla Narasimha Reddy Education Society, suggesting a background of academic dedication in pharmacy education and contributions to pharmaceutical research within India.

8) Education:
Education denotes the process of fostering knowledge and skills. In this setting, the institution's role is emphasized in training future pharmacists and researchers while contributing to the scientific community through studies like the one on Metformin, pivotal for developing competent professionals in the pharmaceutical industry.

9) Relative:
Relative indicates a relationship or comparison between entities. In the context of this research, relative values such as standard deviation or percentage recovery are used to evaluate the precision and accuracy of the analytical methods, providing insights into the consistency of results and the performance of methodologies employed for Metformin analysis.

10) Powder:
Powder refers to a solid material composed of tiny particles. In pharmaceutical contexts, the form and characteristics of powders, such as Metformin, are crucial for formulation and pharmacokinetics. The article discusses powder characteristics when preparing samples for analysis, influencing the method selected and outcomes of the assessment.

11) India:
India serves as the geographical and institutional context for the research. The study highlights contributions from Indian researchers and educational institutions, reflecting the nation's growing pharmaceutical industry and commitment to healthcare advancements, particularly concerning diabetes management through effective drug analysis and validation methodologies.

12) Nalla:
Nalla refers to the name within the educational institution's title. It signifies the Nalla Narasimha Reddy Education Society, underpinning the academic background of the authors, which enhances the credibility of the study and emphasizes the society's mission toward excellence in pharmacy education and research contributions.

13) Observation:
Observation embodies the act of scrutinizing data or phenomena to gain insights. In the research paper, observations relate to recorded data from experiments and analyses of Metformin, crucial for interpreting results and making conclusions about the efficacy and reliability of the developed methods for pharmaceutical analysis.

14) Discussion:
Discussion entails the evaluation and interpretation of research findings. This section synthesizes results, compares them against expectations, and proposes implications for future research or practical applications in formulating Metformin, serving as a vital communicator of knowledge to the scientific community and stakeholders in healthcare.

15) Krishna (Krsna):
Krishna likely refers to one of the authors involved in the research. The inclusion of individual names in academic work is vital as it provides recognition to contributors' roles, expertise, and academic backgrounds, promoting accountability and academic integrity in the published findings surrounding Metformin analysis.

16) Filling (Filled):
Filled pertains to the process of adequately preparing samples or apparatus for experiments. In the research article, it refers to filling volumetric flasks or chromatographic systems with solutions, which is critical for experimental accuracy and subsequent analysis of Metformin, thereby ensuring reliable data collection in the quality assessment.

17) Priya:
Priya is likely one of the contributing authors. The inclusion of author names in research highlights collaboration and shared responsibility for the study's findings related to Metformin, enhancing the work's credibility and showcasing interdisciplinary teamwork among scientists in pharmaceutical research.

18) Pari:
Pari likely refers to another collaborator in the research study. Mentioning all authors supports transparency in academic research, attributing credit to participants according to their contributions to developing and validating analytical methods for Metformin, showcasing the collective effort directed toward meaningful advancements in the field.

19) Line:
Line refers to defined boundaries in scientific research, such as establishing methods or parameters for analysis. A linear relationship in calibration curves denotes reliable measurements of Metformin concentrations, highlighting the significance of accurately interpreting associated data in method validation and quantifying the drug's presence in formulations effectively.

20) Life:
Life, in this context, reflects the importance of drug efficacy in enhancing quality of life for patients, particularly those with diabetes. By developing accurate analytical methods for Metformin, the study contributes to healthcare, ensuring effective management of diabetes and thereby positively impacting patient outcomes and overall public health.

Other Science Concepts:

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Discover the significance of concepts within the article: ‘Method development and validation of metformin by HPLC.’. Further sources in the context of Science might help you critically compare this page with similair documents:

Non-insulin dependent diabetes mellitus, Average weight, Retention time, Limit of detection, Method development, HPLC Method, Calibration curve.

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