HPLC analysis of ascorbic, gallic, and catechin using Box-Behnken.
Journal name: World Journal of Pharmaceutical Research
Original article title: Simultaneous quantitative hplc analysis of ascorbic acid, gallic acid, and catechin in punica granatum, tamarindus indica and prunus domestica using boxbehnken statistical design
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Mohd Amir, Mohd Mujeeb, Sayeed Ahmad, Mohd Akhtar, Kamal YT, Kamran Ashraf
World Journal of Pharmaceutical Research:
(An ISO 9001:2015 Certified International Journal)
Full text available for: Simultaneous quantitative hplc analysis of ascorbic acid, gallic acid, and catechin in punica granatum, tamarindus indica and prunus domestica using boxbehnken statistical design
Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research
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Summary of article contents:
Introduction
The study presented by Mohd Mujeeb et al. focuses on developing a validated reversed-phase high-performance liquid chromatography (HPLC) method for the simultaneous estimation of ascorbic acid, gallic acid, and catechin found in the fruits of Punica granatum (pomegranate), Tamarindus indica (tamarind), and Prunus domestica (plum). The method aims to enhance the quality control of herbal drugs by providing a reliable analytical technique to quantify these antioxidant compounds that are known for their biological significance and potential health benefits against oxidative stress-induced diseases.
Optimization of HPLC Conditions
The researchers optimized the chromatographic conditions for effective separation of the target compounds. Using a C18 reverse phase column, they established a mobile phase consisting of 0.1% glacial acetic acid and acetonitrile in a 70:30 (v/v) ratio, with a flow rate of 1.0 mL/min, resulting in effective separation within a 15-minute run time. The detection was conducted at a wavelength of 254 nm, allowing for symmetrical and well-resolved peaks for ascorbic acid, gallic acid, and catechin, with retention times recorded at 2.163, 3.269, and 6.392 minutes, respectively.
Method Validation and Precision
The validation of the HPLC method was performed according to International Conference on Harmonisation (ICH) guidelines, which evaluated aspects like selectivity, linearity, precision, robustness, accuracy, and limits of detection and quantification (LOD and LOQ). The method exhibited high linearity within specific concentration ranges for each compound, with regression coefficients above 0.995. The precision studies indicated low relative standard deviations (RSD) for both intra-day and inter-day analyses, confirming the method's reliability and consistency.
Robustness of the Method
The robustness of the developed method was assessed using Box-Behnken statistical design, which examined the effects of small changes in temperature, flow rate, and detection wavelength on peak area responses. The findings demonstrated that flow rate and wavelength had significant effects on the peak area for ascorbic acid, gallic acid, and catechin, while temperature inversely affected peak area response. These evaluations ensured that the method is not only precise but also resilient against minor fluctuations in analytical conditions.
Conclusion
In conclusion, the research successfully established a validated HPLC method for the simultaneous determination of key antioxidant compounds in P. granatum, T. indica, and P. domestica. The method proved to be simple, accurate, specific, and cost-effective, with robust analytical capabilities suitable for routine quality control of herbal drugs. Future implications of this work extend to employing this analytical method for identifying and quantifying antioxidant compounds in other herbal extracts, thereby enhancing the understanding of their health benefits and therapeutic potential.
FAQ section (important questions/answers):
What is the main purpose of the HPLC method developed?
The HPLC method aims to simultaneously estimate ascorbic acid, gallic acid, and catechin in Punica granatum, Tamarindus indica, and Prunus domestica for quality control and analysis of these antioxidant compounds.
What are the key components analyzed in this study?
The study focuses on three key components: ascorbic acid, gallic acid, and catechin, which are known for their antioxidant properties and potential health benefits.
How did the researchers validate the HPLC method?
Validation was performed through selectivity, linearity, precision, robustness, accuracy, limits of detection and quantification, adhering to ICH guidelines.
What were the limits of detection for the compounds?
The limits of detection were 0.31 µg/mL for ascorbic acid, 1.53 µg/mL for gallic acid, and 15.16 µg/mL for catechin.
What is the significance of using the Box-Behnken design?
Box-Behnken design was used to assess the robustness of the HPLC method and analyze the effects of temperature, flow rate, and wavelength on the peak area.
What are the findings regarding the antioxidant compounds in the fruits?
The quantification results showed varying concentrations of ascorbic acid, gallic acid, and catechin in the tested fruits, highlighting their potential as antioxidant sources.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “HPLC analysis of ascorbic, gallic, and catechin using Box-Behnken.”. 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 the context of scientific research, 'Table' refers to a structured representation of data in rows and columns, which provides a clear overview of experimental results, such as quantification of compounds in samples. This facilitates easy comparison and analysis, thus enhancing understanding of the findings in research articles.
2) India:
India, a country in South Asia, is significant in this study as it serves as the geographical context for the research. The herbal samples, including Punica granatum and Tamarindus indica, were collected from this region, which is known for its rich biodiversity and traditional use of medicinal plants.
3) Water:
Water is a crucial solvent in the extraction and analysis processes of the compounds studied. As an essential part of the mobile phase in HPLC, it ensures effective separation and quantification of ascorbic acid, gallic acid, and catechin from the herbal samples during the chromatographic analysis.
4) Drug:
In the context of this study, 'Drug' refers to the bioactive compounds derived from herbal sources that exhibit pharmacological activities. Analyzing these compounds is essential for quality control, as they are used in traditional medicine and play a vital role in health and wellness.
5) Delhi:
Delhi is the capital of India and serves as a key location for this research. The fruits of the plants were collected here, highlighting the local biodiversity and the importance of regional flora in herbal medicine, allowing for the assessment of important antioxidant constituents.
6) New Delhi:
New Delhi, as the administrative capital of India, represents a significant urban center where this research was conducted. The studies were facilitated by institutions located in New Delhi, which plays a crucial role in promoting scientific research and development in herbal pharmacology.
7) Tamarindus indica:
Tamarindus indica, commonly known as tamarind, is one of the studied plant species in this research. This fruit is recognized for its nutritional and medicinal properties, particularly its antioxidant activity, making it a valuable component in traditional and modern herbal medicine.
8) Punica granatum:
Punica granatum, known as pomegranate, is another plant investigated in the study. This fruit is celebrated for its high antioxidant properties, including its content of ascorbic acid and other phenolic compounds, which are associated with various health benefits and disease prevention.
9) Relative:
The term 'Relative' in this context refers to the measures used to compare the amounts of different compounds in the study, such as the relative standard deviations (RSD) that gauge the precision of analytical results, indicating the method's reliability in quantifying specific antioxidants.
10) Disease:
'Disease' is a critical term in the discussion of the health benefits of antioxidants highlighted in the research. The study emphasizes how compounds like ascorbic acid, gallic acid, and catechin play a role in combatting oxidative stress related to chronic diseases such as cancer and cardiovascular disorders.
11) Study (Studying):
'Study' refers to the systematic investigation undertaken in this research to explore the simultaneous quantification of antioxidants in specific fruits. This study aims to validate a robust analytical method, contributing to the knowledge base regarding the health benefits of plant-based antioxidants.
12) Activity:
In the context of this research, 'Activity' refers to the biological activities of the compounds analyzed, particularly their antioxidant properties. These activities are crucial for preventing oxidative damage in cells, contributing to the potential therapeutic effects of the studied herbal medicines.
13) Quality:
'Quality' relates to the assessment of herbal ingredients based on their content of active compounds. In this study, quality control is essential to ensure that the levels of ascorbic acid, gallic acid, and catechin meet safety and efficacy standards for therapeutic use.
14) Surface:
'Surface' refers to the methodology applied for evaluating the analytes in this research, particularly in the context of the Box-Behnken design used in robust testing. This term also pertains to the interaction of the compounds with the reverse-phase HPLC column surface during analysis.
15) Human body:
The 'Human body' is the biological context within which the antioxidant activity of the studied compounds operates. Understanding how dietary antioxidants influence health outcomes, particularly their role in mitigating oxidative stress and preventing chronic diseases, is crucial in this research.
16) Pharmacological:
The term 'Pharmacological' relates to the study of how the bioactive compounds found in the analyzed plants affect biological systems. This research aims to elucidate the pharmacological relevance of ascorbic acid, gallic acid, and catechin in the prevention of chronic diseases.
17) Pharmacology:
Pharmacology is the branch of medicine that focuses on the study and effects of drugs. This research contributes to the field by providing insights into the therapeutic potential of plant-derived antioxidants, emphasizing their importance in promoting health and managing diseases.
18) Pomegranate:
Pomegranate is a rich source of antioxidants, particularly ascorbic acid and other polyphenols studied in this research. Its health benefits, including anti-inflammatory and anti-cancer properties, underline the importance of quantifying its bioactive constituents for nutritional and medicinal purposes.
19) Discussion:
'Discussion' is a vital section in scientific literature that interprets the results and places them in the context of existing knowledge. This study provides a discussion on the significance of the findings, the methodology used, and implications for future research on antioxidants.
20) Tamarind:
Tamarind is highlighted in this study as a significant source of bioactive compounds. Its traditional use in medicine and its rich content of antioxidants such as gallic acid demonstrate its importance in dietary practices and its impact on health.
21) Account:
'Account' refers to the detailed reporting of findings in research. In this study, various analytical parameters are accounted for, including precision, accuracy, and linearity, which are crucial in validating the HPLC method used for quantifying the studied compounds.
22) Mineral:
Minerals play a supporting role in the effectiveness of antioxidants and overall health. Although the study focuses on specific antioxidant compounds, the synergistic effect of minerals in diet highlights their importance in enhancing the bioactivity of the studied plant extracts.
23) Family:
'Family' refers to the taxonomic classification of the studied plants, which belong to different botanical families. Understanding the familial relationships among these plants can provide insights into their shared characteristics and potential similarities in the phytochemical profiles.
24) Cancer:
Cancer is a significant focus in this research due to the established link between oxidative stress and cancer development. The study emphasizes how antioxidants can mitigate cellular damage and contribute to cancer prevention, showcasing the health benefits of the tested compounds.
25) Botany:
'Botany' is the scientific study of plants, which provides the foundational knowledge for this research. Understanding plant taxonomy, morphology, and phytochemistry is essential for the identification and classification of the medicinal plants analyzed in this study.
26) Ayus (Āyus, Ayush):
'Ayush' refers to the Department of AYUSH, which promotes traditional medicine practices in India, including Ayurveda, Yoga, and Homeopathy. The study acknowledges the support from this department, indicating the importance of integrating modern scientific research with traditional medicinal knowledge.
27) Diet:
'Diet' is a significant factor affecting human health, as it influences the intake of antioxidants. The study discusses the role of a diet rich in fruits and vegetables as a means of obtaining essential antioxidants necessary for health and disease prevention.
28) Rich (Ṛch):
'Rich' describes the significant abundance of antioxidants present in the examined fruits. This term underscores the nutritional value of Punica granatum, Tamarindus indica, and Prunus domestica, indicating their potential role in a health-promoting diet.
Other Science Concepts:
Discover the significance of concepts within the article: ‘HPLC analysis of ascorbic, gallic, and catechin using Box-Behnken.’. Further sources in the context of Science might help you critically compare this page with similair documents:
Punica granatum, Tamarindus indica, Analytical study, Analytical method, Accuracy, Experimental design, Financial assistance, Quality Control, Antioxidant activity, Chromatogram, Temperature effect, Retention time, Mobile phase, Ascorbic acid, Marker compound, Standard deviation, HPLC Method, Gallic acid, Herbal drug, Department of Pharmacology, Chemical constituent, Recovery studies, Method validation, Limits of detection and quantification, Relative standard deviation, Stock Solution, Chromatographic condition, Selectivity, Reverse phase HPLC, Catechin, Linear regression equation, Pharmacognosy and Phytochemistry, Reverse Phase C18 Column, Health benefit, Chemical component, HPLC fingerprinting.