U-HPLC separation of the indole alkaloid strychnine.
Journal name: World Journal of Pharmaceutical Research
Original article title: U-hplc (ultra-high-pressure liquid chromatography) separation of indole alkaloid strychnine
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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Pasupuleti Sreenivasa Rao, Gangapatnam Subramanayam and Matcha Bhaskar
World Journal of Pharmaceutical Research:
(An ISO 9001:2015 Certified International Journal)
Full text available for: U-hplc (ultra-high-pressure liquid chromatography) separation of indole alkaloid strychnine
Source type: An International Peer Reviewed Journal for Pharmaceutical and Medical and Scientific Research
Doi: 10.20959/wjpr201713-9955
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Summary of article contents:
Introduction
The study focuses on the separation of the indole alkaloid strychnine from the genus Strychnos using Ultra-High-Pressure Liquid Chromatography (UHPLC), marking a significant advancement in chromatographic techniques. This genus is known for its numerous species that possess important medicinal properties, particularly due to the presence of indole alkaloids, which have been utilized in various traditional and modern medical practices. Previous isolation methods, including Reverse Phase High-Performance Liquid Chromatography (RP-HPLC), have yielded valuable insights into the chemical compositions of these compounds; however, this research uniquely highlights the efficiency and effectiveness of UHPLC in facilitating rapid analysis with minimal solvent use.
Significance of UHPLC
UHPLC stands out as a state-of-the-art analytical technique that offers significant advantages over traditional HPLC methods. The study emphasizes that UHPLC enables quicker analyses and shorter run times, resulting in lower solvent consumption and reduced usage of laboratory equipment. The research establishes a highly efficient methodology for the separation of strychnine, achieving optimal conditions for effective peak resolution. The ability to complete the separation of strychnine in under six minutes demonstrates the potential of UHPLC for both qualitative and quantitative analyses in pharmacognosy.
Methodological Innovations
A detailed methodology underpins the successful separation of strychnine using the developed UHPLC technique. The parameters for the mobile phase were carefully optimized, utilizing a methanol-water solvent mix in a 20:80 ratio at a flow rate of 0.75 ml/min, while monitoring the eluate at 254 nm. The study succeeds in establishing the reproducibility of peak identification at a consistent retention time of 5.3 minutes, showcasing the robustness of the method. This careful calibration and optimization of chromatographic conditions represent a crucial innovation in the extraction and analysis of indole alkaloids.
Analytical Outcomes
The quantitative analysis of strychnine concentration revealed a strong linear correlation across tested ranges from 5 µg to 1 mg. Calibration curves indicated a high degree of reliability, as demonstrated by the regression equation (y = 2448.01x + 878906.6) with a correlation coefficient of 0.99. This level of precision indicates that the UHPLC method not only improves upon previous techniques but offers valid and reproducible results essential for further pharmacological and toxicological studies related to strychnine and its implications in traditional and modern medicine.
Conclusion
In conclusion, the study successfully establishes a simple, efficient, and repeatable UHPLC method for the identification and quantification of indole alkaloids, particularly strychnine. By harnessing the advantages of UHPLC, the research addresses the limitations of conventional techniques, providing a reliable platform for future investigations into the pharmacological potentials of Strychnos species. This work not only contributes to the field of medicinal chemistry but also emphasizes the importance of modern chromatographic techniques in exploring natural products with therapeutic applications.
FAQ section (important questions/answers):
What is the focus of the study conducted by Rao et al.?
The study focuses on the separation and analysis of the indole alkaloid strychnine using ultra-high-pressure liquid chromatography (UHPLC) for the first time, highlighting its effectiveness in rapid qualitative and quantitative analysis.
What are the main alkaloids studied in the Strychnos genus?
The main alkaloids in the Strychnos genus studied are strychnine and brucine, which are noted for their medicinal properties but can also be toxic in high doses.
How does UHPLC differ from traditional HPLC methods?
UHPLC is an updated version of HPLC that allows for quicker analysis with shorter run times, better column equilibration, and is particularly suitable for method development in the separation of compounds.
What was the optimal mobile phase composition used in the study?
The optimal mobile phase composition used in the study was a mixture of methanol and water in a 20:80 ratio, facilitating effective separation of strychnine at a flow rate of 0.75 ml/min.
What was the retention time for strychnine during UHPLC?
The retention time for strychnine during the UHPLC analysis was consistently noted at 5.3 minutes, unaffected by varying concentrations of the compound.
What significance does this study have for pharmaceutical research?
This study is significant as it establishes a reliable UHPLC method for identifying and quantifying strychnine, potentially aiding in the standardization and quality control of this important alkaloid in pharmaceutical applications.
Glossary definitions and references:
Scientific and Ayurvedic Glossary list for “U-HPLC separation of the indole alkaloid strychnine.”. 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) Study (Studying):
Studying refers to the investigation and analysis of a subject or phenomenon to gain knowledge or insight. In this article, studying the separation of strychnine through U-HPLC highlights the relevance of advanced analytical techniques in botany and pharmacology, thereby aiding in understanding the chemical properties of medicinal plants.
2) Genu:
Genu is likely a typographical error for 'genus,' which refers to a rank in biological classification, one of several levels of taxonomy. The genus Strychnos in this context represents a group of species known to produce indole alkaloids, including the well-studied strychnine. This classification aids in the understanding of relationships among different plant species.
3) Species:
Species is a fundamental unit of biological classification representing a group of organisms that can interbreed and produce fertile offspring. The article discusses various species within the Strychnos genus that are valuable for medicinal uses due to their alkaloid composition. Understanding species diversity is crucial for pharmacological and ecological studies.
4) Purification:
Purification is the process of isolating a compound or substance from a mixture, enhancing its purity for further use or analysis. In this study, it relates to scientifically separating strychnine from other constituents in plant extracts to understand its properties and effects. High purity levels ensure accurate evaluations of biological activity.
5) Disease:
Disease refers to the pathological condition of a living organism, often characterized by specific symptoms and signs. The research emphasizes the traditional medicinal values of Strychnos species in treating various diseases, highlighting the potential therapeutic applications of indole alkaloids like strychnine in modern medicine and pharmacology.
6) Drug:
A drug is a substance used to diagnose, cure, or prevent a disease, or to enhance physical or mental well-being. The article indirectly pertains to drug development by discussing compounds derived from Strychnos species. Understanding natural drugs like strychnine may lead to advancements in developing effective treatments against various ailments.
7) Medicine:
Medicine is the science and practice of diagnosis, treatment, and prevention of disease. The paper highlights the importance of studying alkaloids from Strychnos as a potential source of medicinal compounds. This knowledge contributes to the field of pharmaceutical development and offers insights into traditional healing practices.
8) Cancer:
Cancer is a disease characterized by uncontrolled growth of abnormal cells. The article notes that alkaloids from Strychnos species have anti-cancer properties. This information is crucial for developing targeted therapies and underscores the importance of exploring natural compounds as sources for effective cancer treatments.
9) India:
India is a country in South Asia noted for its diverse flora and rich traditional medicinal practices. The study is situated within Indian contexts, particularly focusing on Strychnos species found in the region. Its position parallels the global exploration of plant-derived alkaloids for medicinal applications.
10) Water:
Water is a vital solvent in scientific research and plays a crucial role in various biological processes. In the paper, water is used as a component in the mobile phase for UHPLC analyses. Its presence is significant for solvent interactions in the chromatographic separation of compounds like strychnine.
11) Antibiotic (Antibacterial):
Anti-bacterial compounds inhibit the growth of bacteria. The research acknowledges that indole alkaloids from the Strychnos species possess antibacterial properties, which can inform the development of new antibacterial drugs. This study highlights the potential of natural plant compounds in combating bacterial infections.
12) Cosmopolitan:
Cosmopolitan refers to species that are widespread and can thrive in diverse environments across the globe. The Strychnos genus is described as cosmopolitan, suggesting its evolutionary adaptability, which is important for researchers as they study its various medicinal properties in a wide range of ecological settings.
13) Science (Scientific):
Scientific pertains to methods and approaches based on empirical evidence and standardized procedures. The article utilizes scientific techniques like U-HPLC to analyze strychnine, demonstrating the importance of rigorous methodologies in pharmacological research, which helps in establishing reliability and credibility in findings.
14) Narayana (Nara-ayana, Nara-yana):
Narayana refers to Narayana Medical College and Hospitals in India, associated with advanced research in clinical and pharmaceutical sciences. The institution plays a key role in the study, underpinning the local commitment to researching valuable natural medicines derived from indigenous plants, thus enhancing local healthcare innovation.
15) Relative:
Relative pertains to the systematic relationships or comparisons among entities. In the context of this study, it might refer to the relative purity of the strychnine or the relative efficacy of alternative chromatographic methods in assessing the alkaloids, which is crucial for accurate pharmacological applications.
16) Purity:
Purity signifies the absence of impurities in a substance, critical for accurate scientific analysis. In this study, achieving high purity of strychnine through UHPLC enhances reliability in subsequent qualitative and quantitative evaluations, ensuring valid interpretations and potential therapeutic applications of the compound.
17) Snake poison (Snake venom):
Snake poison refers to toxins released by snakes as a defense mechanism or when hunting prey. Strychnine is noted for its toxicological properties, akin to snake venom. The article likely discusses its medicinal applications as an antidote for snake bites, demonstrating the dual nature of alkaloids in nature.
18) Phytochemical:
Phytochemical denotes chemical compounds produced by plants, often possessing biological activity. The research focuses on the phytochemicals, especially indole alkaloids from Strychnos species, underscoring their importance in drug discovery and development to combat various health conditions and contribute to knowledge of medicinal plants.
19) Discussion:
Discussion in scientific literature refers to analysis and interpretation of findings. The article engages in a thorough discussion of the methods and results of strychnine analysis, contributing to the broader understanding of alkaloidal research, drug development, and possible therapeutic applications for these compounds in medicine.
20) Tirupathi:
Tirupathi is a city in India known for its religious significance and educational institutions. It plays a role in the study as the location of Sri Venkataeswara University, which is involved in researching pharmacological properties of indigenous plants like those in the Strychnos genus, enhancing local education.
21) Flavonoid:
Flavonoid refers to a diverse group of phytonutrients with antioxidant properties found in various plants. While not the main focus in this study, flavonoids have been explored for their health benefits. Understanding these compounds in conjunction with alkaloids can provide comprehensive insights into the synergistic effects of plant-based medicines.
22) Toxicity:
Toxicity refers to the degree to which a substance can cause harm to an organism. In the context of strychnine, its toxicity is highlighted, necessitating careful handling and understanding of its pharmacological effects. This is significant for distinguishing its therapeutic potentials from its harmful side effects.
23) Activity:
Activity in pharmacological terms refers to the effectiveness of a drug in producing a biological effect. The article investigates the biological activity of strychnine, focusing on its separation methods. This highlights the importance of understanding and quantifying the activity of alkaloids in therapeutic contexts.
24) Channel:
Channel in a scientific context might refer to pathways or means through which substances are analyzed or transported. The UHPLC methods used in this study employ channels to efficiently separate compounds, demonstrating the significance of technology and methodology in obtaining reliable results in chemical research.
25) Quality:
Quality pertains to the standard or degree of excellence of a substance. The study focuses on quality analysis of the indole alkaloid strychnine through rigorous UHPLC methods, ensuring that the derived data accurately reflects its characteristics, which is crucial for pharmaceutical development.
26) Zoology:
Zoology is the scientific study of animals and their biology. Although not the primary focus of this article, it may relate to the Division of Animal Biotechnology mentioned, signifying interdisciplinary research where understanding animal physiology can complement studies of naturally derived compounds for medical applications.
27) Company:
Company in this context could refer to the local or commercial entities providing chemicals and materials for research. Such partnerships are essential in scientific investigations, as collaboration enhances access to necessary resources for studies like those involving chromatographic techniques and alkaloid purification.
28) Andhra (Amdhra):
Andhra refers to Andhra Pradesh, a state in India, significant for its rich biodiversity and traditional medicinal practices. The study's geographical context highlights the state's contribution to the exploration of local flora, like Strychnos species, which have potential medicinal applications in combating diseases.
29) Family:
Family is a taxonomic rank used in the classification of organisms; for instance, the family Loganiaceae includes the genus Strychnos. Understanding the family relationships helps researchers categorize and better study the properties and uses of plant species, particularly in studying natural products for medicinal purposes.
30) Animal:
Animal refers to organisms in the kingdom Animalia. In the context of this article, it underscores the relevance of studying animal models in pharmacological research, particularly when examining the effects of plant-derived compounds like strychnine, contributing to understanding drug actions and potential toxicities.
31) Doubt:
Doubt refers to uncertainty or lack of confidence in a particular outcome or result. The article mentions prior studies where TLC provided ambiguous results concerning the alkaloid separation, thus underscoring the reliability of advanced techniques like UHPLC in yielding clearer and more definitive data.
32) Field:
Field refers to a specific area of study or expertise. In this article, the field encompasses pharmacology, botany, and phytochemistry, highlighting interdisciplinary approaches in researching plant-derived compounds for medicinal uses and the advancement in analytical techniques for assessment of these natural biologically active substances.
33) Fight:
Fight signifies the struggle against diseases or health challenges. The article emphasizes the potential role of Strychnos-derived alkaloids in combating various diseases, suggesting that such natural compounds can contribute to innovative therapeutic strategies in the ongoing fight against ailments affecting human health.
34) Bail:
Bail may refer to a specific species of Strychnos or could be a typographical error. In the context of the article, if it refers to a species, it emphasizes the biological diversity within the Strychnos genus, which is essential in exploring the pharmacological potential of various indole alkaloids for therapeutic applications.