Product Research Research Articles

Core publications in drug discovery and natural product research

Natural products have long been a cornerstone of drug discovery, providing diverse and biologically relevant chemical scaffolds. This work aims to guide newcomers to natural product research and, specifically, drug discovery by presenting a curated list of 30 key publications selected through an international survey of experts and critical evaluation by the authors. The selected works span textbooks, review articles, and original research papers, covering various aspects of natural product research, including chemistry, pharmacology, analytical sciences, emerging open science, and computational approaches. We discuss historical milestones in natural product drug discovery, highlighting the specific contributions of the U.S. National Cancer Institute in developing anticancer and anti-HIV agents. The present work also addresses current challenges and innovations in the field, emphasizing the importance of data quality, interdisciplinary collaboration, and the integration of artificial intelligence. By providing this carefully selected reading list and accompanying analysis, we aim to offer a comprehensive yet accessible entry point for researchers new to natural product-based drug discovery and highlight future directions and opportunities in this dynamic field.

In Silico Studies as Support for Natural Products Research

In Silico Studies as Support for Natural Products Research

Using models and maps to inform Target Product Profiles and Preferred Product Characteristics: the example of Wolbachia replacement.

The global prevalence of diseases transmitted by Aedes aegypti mosquitoes, such as dengue, Zika and Yellow Fever, is increasing, but development of promising new mosquito control technologies could reverse this trend. Target Product Profiles (TPPs) and Preferred Product Characteristics (PPCs) documents issued by the World Health Organization can guide the research and development pathways of new products and product combinations transitioning from proof of concept to operational use. We used high resolution global maps of the case and economic burden of dengue to derive programmatic cost targets to support a TPP for Wolbachia replacement. A compartmental entomological model was used to explore how release size, spacing and timing affect replacement speed and acceptability. To support a PPC for a hybrid suppress-then-replace approach we tested whether Wolbachia replacement could be achieved faster, more acceptably or at a lower cost if preceded by a mosquito suppression programme. We show how models can reveal trade-offs, identify quantitative thresholds and prioritise areas and intervention strategies for further development. We estimate that for Wolbachia replacement to be deployable in enough areas to make major contributions to reducing global dengue burden by 25% (in line with 2030 WHO targets), it must have the potential for cost to be reduced to between $7.63 and $0.24 (USD) per person protected or less. Suppression can reduce the number of Wolbachia mosquitoes necessary to achieve replacement fixation by up to 80%. A hybrid approach can also achieve fixation faster and potentially improve acceptability, but may not justify their cost if they require major new investments in suppression technologies. Here we demonstrate the value dedicated modelling can provide for interdisciplinary groups of experts when developing TPPs and PPCs. These models could be used by product developers to prioritise and shape development decisions for new Wolbachia replacement products.

Targeted discovery of polyketides with antioxidant activity through integrated omics and cocultivation strategies.

Fungi generate a diverse array of bioactive compounds with significant pharmaceutical applications. However, the chemical diversity of natural products in fungi remains largely unexplored. Here, we present a paradigm for specifically discovering diverse and bioactive compounds from fungi by integrating genome mining with building block molecular network and coculture analysis. Through pangenome and sequence similarity network analysis, we identified a rare type I polyketide enzyme from Penicillium sp. ZJUT-34. Subsequent building block molecular network and coculture strategy led to the identification and isolation of a pair of novel polyketides, (±)-peniphenone E [(±)-1], three known polyketides (2-4), and three precursor compounds (5-7) from a combined culture of Penicillium sp. ZJUT-34 and Penicillium sp. ZJUT23. Their structures were established through extensive spectroscopic analysis, including NMR and HRESIMS. Chiral HPLC separation of compound 1 yielded a pair of enantiomers (+)-1 and (-)-1, with their absolute configurations determined using calculated ECD methods. Compound (±)-1 is notable for its unprecedented structure, featuring a unique 2-methyl-hexenyl-3-one moiety fused with a polyketide clavatol core. We proposed a hypothetical biosynthetic pathway for (±)-1. Furthermore, compounds 2, 5, and 6 exhibited strong antioxidant activity, whereas (-)-1, (+)-1, 3, and four exhibited moderate antioxidant activity compared to the positive control, ascorbic acid. Our research demonstrates a pioneering strategy for uncovering novel polyketides by merging genome mining, metabolomics, and cocultivation methods. This approach addresses the challenge of discovering natural compounds produced by rare biosynthetic enzymes that are often silent under conventional conditions due to gene regulation.IMPORTANCEPolyketides, particularly those with complex structures, are crucial in drug development and synthesis. This study introduces a novel approach to discover new polyketides by integrating genomics, metabolomics, and cocultivation strategies. By combining genome mining, building block molecular networks, and coculturing techniques, we identified and isolated a unique polyketide, (±)-peniphenone E, along with three known polyketides and three precursor compounds from Penicillium sp. ZJUT-34 and Penicillium sp. ZJUT23. This approach highlights the potential of using combined strategies to explore fungal chemical diversity and discover novel bioactive compounds. The successful identification of (±)-peniphenone E, with its distinctive structure, demonstrates the effectiveness of this integrated method in enhancing natural product discovery and underscores the value of innovative approaches in natural product research.

Microgreens market and prospects for their use as a functional ingredient in food production

Currently, the microgreens market is actively growing all over the world. Due to the fragmentation of this market, high competition from manufacturers, growing consumer demand, and the high nutrient status of microgreens, research and development of new food products with microgreens as a functional ingredient are promising. The paper presents the main trends in the global microgreens market and discusses aspects of using microgreens as a functional ingredient in food products.

Isolation and Characterization of Stigmasterol from Psidium guajava Leaves: A Promising Bioactive Compound with Therapeutic Potential

This study aimed to isolate and characterize bioactive compounds from the leaves of Psidium guajava Linn. The hydroethanolic extract yielded 32% (80 g) from the crude material, which was subjected to column chromatography for fractionation. Structural elucidation of the isolated compound was performed using 1D and 2D Nuclear Magnetic Resonance (NMR) spectroscopy, including COSY, HSQC, and HMBC experiments. The spectral data confirmed the presence of stigmasterol, characterized by its distinct chemical shifts and long-range correlations between carbon and proton atoms. Stigmasterol was isolated as a white, crystalline solid and identified by its unique physio-chemical properties, such as solubility in organic solvents, a melting point of 163-164°C, and molecular formula C29H48O. This compound is known for its significant biological activities, including antibacterial, antimalarial, anti-inflammatory, antioxidant, and anticancer effects. By employing advanced techniques like 1D and 2D NMR spectroscopy, the study provides precise structural elucidation of stigmasterol, contributing to natural product research and reinforcing the medicinal potential of guava leaves as a source of pharmacologically active compounds. This adds to the existing knowledge of the plant’s bioactive components and their applications in medicinal chemistry

Investigation on taxonomy, secondary metabolites and antibacterial activity of Streptomyces sediminicola sp. nov., a novel marine sediment-derived Actinobacteria.

Marine actinomycetes, especially Streptomyces, are recognized as excellent producers of diverse and bioactive secondary metabolites on account of the multiplicity of marine habitations and unique ecological conditions, which are yet to be explored in terms of taxonomy, ecology, and functional activity. Isolation, culture and genome analysis of novel species of Streptomyces to explore their potential for discovering bioactive compounds is an important approach in natural product research. A marine actinobacteria, designated strain SCSIO 75703T, was isolated, and the potential for bioactive natural product discovery was evaluated based on genome mining, compound detection, and antimicrobial activity assays. The phylogenetic, phenotypic and chemotaxonomic analyses indicate that strain SCSIO 75703T represents a novel species in genus Streptomyces, for which the name Streptomyces sediminicola sp. nov. is proposed. Genome analysis revealed the presence of 25 secondary metabolite biosynthetic gene clusters. The screening for antibacterial activity reveals the potential to produce bioactive metabolites, highlighting its value for in-depth exploration of chemical constituents. Seven compounds (1-7) were separated from the fractions guided by antibacterial activities, including three indole alkaloids (1-3), three polyketide derivatives (4-6), and 4-(dimethylamino)benzoic acid (7). These primarily antibacterial components were identified as anthracimycin (4), 2-epi-anthracimycin (5) and β-rubromycin (6), presenting strong antibacterial activities against Gram-positive bacteria with the MIC value ranged from 0.125 to 16μg/mL. Additionally,, monaprenylindole A (1) and 3-cyanomethyl-6-prenylindole (2) displayed moderate inhibitory activities against α-glucosidase with the IC50 values of 83.27 and 86.21μg/mL, respectively. Strain SCSIO 75703T was isolated from marine sediment and identified as a novel species within the genus Streptomyces. Based on genomic analysis, compounds isolation and bioactivity studies, seven compounds were identified, with anthracimycin and β-rubromycin showing significant biological activity and promising potential for further applications.

The impact of women in natural product research: a legacy of innovation.

The impact of women in natural product research: a legacy of innovation.

Li Nationality Intangible Cultural Heritage in the Development of Cultural and Creative Products Research

As one of the main indigenous nationalities in Hainan Island, the unique intangible cultural heritage of the Li nationality is an important part of the multicultural treasure house of the Chinese nation. This paper aims to explore the li intangible culture in the product development application and research, through the analysis of li traditional spinning weaving techniques, rattan art, li silver and other intangible cultural connotation and artistic characteristics, put forward the use of modern design concept and technology for product development strategy, in order to protect and inheriting li intangible culture provide new ideas and methods.

Dedicated low-cost electronic ocarina for people with COPD: Rationale and prototype development

ABSTRACT Introduction Chronic obstructive pulmonary disease (COPD) is a condition characterized by airflow obstruction which can lead to physical and psychosocial challenges, with a higher prevalence in developing countries. Non-pharmacological interventions for this group of people include pulmonary rehabilitation as well as music interventions. Although music technology was recommended to adjust to the user’s breathing capacity for music-making and to gather information about their performance, music therapy trials used acoustic wind instruments for their rehabilitation. This interdisciplinary project aims to develop a dedicated electronic wind instrument for people with COPD. Method This project follows a systematic prototype methodology that includes product research, instrument development, and user validation. Based in Brazil, our team has decided on an electronic ocarina based on its cultural features, its popularity as a digital application, and its potential for respiratory training. Unlike existing technologies, the prototype intends to be dedicated and a low-cost solution. Results The development process includes decision making about the appropriate mouthpiece, sensors, Arduino, pushbuttons, 3-D printing projects, building a MIDI interface and a dedicated software. This is an ongoing project, and our team is in the MIDI interface development stage. We aim to run the first hardware trial with users so that we can integrate their feedback into the functional prototype and the dedicated software for the following trials. Discussion This project’s strengths rely on the innovation of building viable electronic wind instruments that can facilitate self-management of people with COPD in developing countries, where access to healthcare can be challenging.

Consensus Statement on the Outcome of the European Herbal Health Products Summit - Which Way Forward?

Herbal medicinal products are a vital part of the healthcare system in Europe and beyond. Being predominantly sold as non-prescription medicines in pharmacies, they are very popular with patients, physicians, and pharmacists and are therefore an important part of self-medication. Interest in this sector has recently gained momentum, reflecting the ongoing revision of the general pharmaceutical legislation and the recent discussion on nutrition and health claims on foods based on the implementation report of Regulation (EC) No 1924/2006 by the European Parliament 1. Therefore, on 20th February 2024, the Society for Medicinal Plant and Natural Product Research (GA), in collaboration with the German Pharmaceutical Industry Association (BPI) and the German Medicines Manufacturers' Association (BAH, now Pharma Deutschland), hosted an in-person summit in Brussels entitled 'European Herbal Health Products Summit - Which way forward?'. The summit featured a wide range of speakers, including policymakers, regulatory authorities, industry representatives, and academic experts. It was divided into several sessions covering topics such as the future and relevance of herbal medicinal products in the EU, the revision of the EU pharmaceutical legislation, and the resulting impact on herbal medicinal products. Furthermore, the discussions delved into the "Health Claims Regulation" - the European Parliament's implementation report and the related regulatory challenges of herbal medicinal products at an EU level. This consensus paper summarises the current status and provided recommendations to pave the way for future strategies to ascertain the continued use of herbal medicinal products as an important therapeutic option for patients.

Detection of Inverse Stable Isotopic Labeling in Untargeted Metabolomic Data.

Stable isotopic labeling is a powerful tool for determining the biosynthetic origin of metabolites and for discovering natural products that incorporate precursors of interest. When isotopically substituted precursors are not available commercially or synthetically, inverse stable isotopic labeling (InverSIL) is a useful alternative. With InverSIL, an organism is grown on an isotopically substituted medium and then fed precursors of natural isotopic abundance which can be tracked by mass spectrometry, thereby bypassing issues with precursor availability. Currently, there is no automated way to identify precursor incorporation in untargeted metabolomic data using InverSIL without specifying an expected change in the mass-to-charge ratio of metabolites that have incorporated the precursor. This makes it difficult to identify unknown natural products that may incorporate portions of precursors of interest using new biochemistry or to rapidly identify incorporation of multiple precursors into different metabolites simultaneously. To address this, we developed a new, robust workflow for the automated identification of inverse labeling in untargeted metabolomic data. We then use this method to identify metabolites that incorporate para-aminobenzoic acid and different portions of l-methionine, including in the same sample, and in the process discover the likely biosynthetic origin for the C-7 and C-9 methyl groups of the pterin portion of dephosphotetrahydromethanopterin, a C1 transfer coenzyme used by methylotrophic bacteria. This workflow can be applied in the future to streamline the use of the versatile InverSIL approach for natural product and metabolism research.

Harnessing Endophytic Fungi from Terminalia chebula: A Dual Approach to Antimicrobial and Phytochemical Exploration

Endophytic fungi isolated from Terminalia chebula exhibit potent antimicrobial activity, showcasing their potential to develop novel therapeutic agents. Additionally, these fungi produce diverse phytochemicals, which may contribute to their medicinal properties and offer new avenues for natural product research. This research study investigated the endophytic fungi isolated from T. chebula and assessed their antimicrobial and phytochemical potential. A total of 28 endophytic fungal species were isolated from leaf and bark tissues. Antimicrobial assays revealed significant inhibitory effects against pathogenic fungi Candida albicans - MTCC 183 & Saccharomyces cerevisiae - MTCC 174 and bacteria Escherichia coli - MTCC 2939, & Bacillus subtilis - MTCC 1305, with endophytic Alternaria sp. exhibiting maximum inhibition zone for antifungal activity and endophytic Aspergillus flavus demonstrating potent activity for their antibacterial analysis. A wide array of phytochemical compounds like alkaloids, flavonoids, phenols, steroids, terpenoids, tannin, saponin, etc, were observed whereas, phytochemical screening indicated the presence of alkaloids and flavonoids in all selected endophytes. These findings suggest that T. chebula harbors a rich endophytic community with the potential for producing novel antimicrobial compounds.

Enhancing Patient Safety Through an Integrated Internet of Things Patient Care System: Large Quasi-Experimental Study on Fall Prevention.

The challenge of preventing in-patient falls remains one of the most critical concerns in health care. This study aims to investigate the effect of an integrated Internet of Things (IoT) smart patient care system on fall prevention. A quasi-experimental study design is used. The smart patient care system is an integrated IoT system combining a motion-sensing mattress for bed-exit detection, specifying different types of patient calls, integrating a health care staff scheduling system, and allowing health care staff to receive and respond to alarms via mobile devices. Unadjusted and adjusted logistic regression models were used to investigate the relationship between the use of the IoT system and bedside falls compared with a traditional patient care system. In total, 1300 patients were recruited from a medical center in Taiwan. The IoT patient care system detected an average of 13.5 potential falls per day without any false alarms, whereas the traditional system issued about 11 bed-exit alarms daily, with approximately 4 being false, effectively identifying 7 potential falls. The bedside fall incidence during hospitalization was 1.2% (n=8) in the traditional patient care system ward and 0.1% (n=1) in the smart ward. We found that the likelihood of bedside falls in wards with the IoT system was reduced by 88% (odds ratio 0.12, 95% CI 0.01-0.97; P=.047). The integrated IoT smart patient care system might prevent falls by assisting health care staff with efficient and resilient responses to bed-exit detection. Future product development and research are recommended to introduce IoT into patient care systems combining bed-exit alerts to prevent inpatient falls and address challenges in patient safety.

B-245 Novel monoclonal antibodies validated specifically for immunohistochemistry by performance assessment on tissue microarrays

Abstract Background Immunohistochemistry (IHC) is a laboratory immunoassay that is a pillar of diagnostic pathology, particularly in the identification and characterization of neoplasms and other diseases/disorders. Though performed routinely, rigorous SOP protocols must be determined empirically for each molecular target, beginning with antibody selection. Both traditional polyclonal and hybridoma-based monoclonal antibodies have been employed in the past, though monoclonal, and particularly recombinant monoclonal, antibodies are preferred. However, any reagent used for diagnostic purposes must be meticulously validated to minimize, if not eliminate, erroneous interpretations of IHC assays. Thus, the extent of validation is the pivotal aspect for establishing an antibody as a reliable diagnostic tool for IHC applications. Methods GeneTex has assembled an extensive catalog of rabbit and mouse monoclonal antibodies validated specifically for IHC. These antibodies are tested on both human normal tissue and cancer tissue microarrays, with upwards of 76 normal and numerous tumor tissues included. Images of the staining are available for all the tested tissues. This broad spectrum of tissues allows a researcher to assess the performance of the antibody on known positive and negative samples, thereby minimizing the risk of false positive or false negative readouts (Gown, 2016). In addition, the data images are of sufficient resolution to determine proper cellular localization of the signal. This special catalog of IHC-optimized reagents are listed under the “HistoMAX” designation. New antibodies will be added continuously, many of them being fully recombinant rabbit monoclonals manufactured using a multi-parameter FACS-based methodology to select antigen-specific IgG+ memory B cells from an immunized rabbit with subsequent cloning of the matched IgG heavy and light chains (Starkie et al., 2016). These recombinant antibodies will be validated following the same approach described above. Results Presently more than 130 mouse or rabbit monoclonal antibodies are available in the HistoMAX line of IHC-validated products and are directed against a spectrum of important targets. This includes antibodies for the commonly analyzed cytokeratins 5, 6, 7, 18, 19, and 20, among many others. Antibodies for CTLA4 and PD-L1 headline products for immunology and immunotherapy research, while other reagents are directed against various cadherins. Hormone receptor antibodies are also represented, with antibodies against estrogen receptor alpha, progesterone receptor, Her2/ErbB2, and the androgen receptor. There are also antibodies against various key “Cluster of Differentiation” (CD) proteins as well as other membrane proteins such as ACE2. Many antibodies are relevant to the study of human malignancies, such as CD66e/CEACAM5/CEA, MSH6, p63, MUC1, Collagen IV, EpCAM, PAX6, PAX8, GP2, TRIM29, S100, and GAD65. All antibodies were evaluated as described above. Conclusions Antibodies are application-specific reagents, and the best antibodies for IHC are validated through systematic staining of a multitude of normal and abnormal tissues. This is the only way that an antibody’s performance for IHC can be critically assessed. The HistoMAX reagents consist only of those monoclonal antibodies that have generated expected signals, or the lack of a signal, in a wide range of tissues in a manner consistent with the literature.

Phytochemical profiling, molecular docking, and ADMET evaluation of essential oils from Anaphalis busua and Anaphalis margaritacea in Uttarakhand's Himalayan Terrain

BackgroundThe study focuses on a comparative analysis of essential oils derived from two species of the Anaphalis genus, namely Anaphalis busua and Anaphalis margaritacea, collected at varying elevations in the hilly terrain of Uttarakhand. The investigation aims to explore the phytochemical composition, conduct in silico molecular docking studies, and evaluate ADMET properties of the essential oils to understand their potential bioactivity. MethodsEssential oils were extracted from the whole plants using the hydrodistillation method, and GC–MS was employed for compound identification. A total of 31 compounds were identified in A. busua and 32 in A. margaritacea, constituting 87.86 % and 87.83 % of the total composition, respectively. Major compounds included isocaryophyllene, caryophyllene oxide, geranyl-α-terpinene, α-pinene, γ-muurolene, δ-cadinene, humulene, and copaene. ResultsMolecular docking studies were conducted using PyRx software on ten notable compounds as ligands, targeting cyclooxygenase-2 (COX-2) and human peroxiredoxin 5 to assess antioxidant and anti-inflammatory activities. Isocaryophyllene, with concentrations of 14.07 % (A. busua) and 17.82 % (A. margaritacea), exhibited significant binding affinities of -5.7 and -7.2 kcal/mol against 1HD2 and 4COX, respectively. ConclusionThe research provides valuable insights into the chemical composition of essential oils from Anaphalis plants and validates their bioactive potential through in silico analysis. Isocaryophyllene emerges as a key compound with noteworthy binding affinities, emphasizing its importance in the medicinal and pharmacological applications of essential oils derived from A. busua and A. margaritacea. The results enhance our understanding of the therapeutic potential of Anaphalis essential oils, opening avenues for future research in natural products and drug discovery.

A spectroscopy-based proof-of-concept (POC) for developing loading of pathogen analyzer (LOPA) for dairy products

Detection of bacterial contamination in dairy products of daily use is a challenge worldwide. We have utilized Methylene Blue Reduction Test (MBRT) for quantification of the microbial count in dairy products (milk) and developed a proof-of-concept (POC) based on this for in-filed applications. In this study, we have used pasteurized milk contaminated with model bacteria Escherichia coli, and Staphylococcus aureus for the calibration and validation of the developed POC. The conversion of MB to Leuco-MB i.e., the colorimetric change due to the reduction of MB to Leuco-MB in presence of microbes has been utilized as the tool to detect presence of microbes in milk. The absorbance peak for methylene blue (MB) at 664 nm decreases significantly in presence of microbes and the blue color becomes faded. In our study, we have employed methylene blue (MB) discolouration phenomenon to estimate the microbial count in milk samples using our developed spectroscopy based POC. The limit of detection (LOD) and the limit of quantitation (LOQ) of the POC were found to be 0.32 CFU/mL and 0.97 CFU/mL. The end users of the developed POC are primarily those involved in the production, processing, testing, regulation, and research of dairy products to ensure they meet safety standards and protect public health. These include retailers, dairy farmers, dairy processors, quality control laboratories, regulatory agencies and research institutions. In our experiment, we have observed a significant change in MB absorption in the milk contaminated with microbes. The indigenously developed sensor strips designed for the working of the POC turn to colorless Leuco-MB compared to milk without the microbes. The analysis of the strips has been measured in the developed device.

A dynamic evolutionary game to discourage enterprise “greenwashing”

To meet the increasing consumer demand for green products and services, governments have increased their regulation of enterprise production behaviors so that enterprises may choose “greenwashing” to obtain the premium of green products at the lowest cost under government regulation. The increase of greenwashing hinders green innovation by enterprises. To quantify this problem and seek solutions to promote enterprises cleaner production and sustainable development, we established a tripartite evolutionary game model for enterprises, consumers, and governments. Using the model, we analyzed the stability of their strategy combinations through simulations and explored the impact of some parameters on the strategy selection. We found that (1) the combination of enterprises adopting true green, consumers buying green products, and weak government regulation was the most environmentally friendly and low-cost strategy solution in the tripartite game system. (2) An increase in government penalties or subsidies to enterprises, as well as a reduction of the extra cost of producing green products, can improve the probability that enterprises will adopt cleaner production. Based on the results, the government should define reasonable subsidies and penalties, guide enterprises to assume their social responsibility, and stimulate consumer green demand. Enterprises should improve their moral qualities, actively engage in green innovation, and increase investment in green product research to guide the game system towards the ideal state.

The National Center for Natural Products Research (NCNPR) at 30: A Legacy of Pioneering Research in Natural Products and Dietary Supplements

Since its establishment in 1994, the National Center for Natural Products Research (NCNPR) at the University of Mississippi has made notable contributions to the field of natural product research, coinciding with the passage of the Dietary Supplement Health and Education Act. Over the past three decades, the Center has focused on studying plants, herbs, and other natural materials for applications in medicine, agriculture, and nutraceuticals, particularly in the area of botanical dietary supplements. NCNPR scientists have been actively engaged in developing and improving quality control measures to help ensure the safety of dietary supplements in response to a growing market. The Center’s research efforts have led to its designation as a U.S. Food and Drug Administration Center of Excellence, reflecting its role in advancing scientific understanding of natural products. Through collaborations with various stakeholders and regulators, NCNPR has contributed to shaping the regulatory landscape for botanical dietary supplements, highlighting both their potential health benefits and associated risks, such as product adulteration. The Center’s influence is also evident internationally, as demonstrated by its annual International Conference on the Science of Botanicals, which will mark its 26th year in April 2025. This overview outlines NCNPR’s role in supporting research, regulation, and safety in the natural products field.

Development and Validation of a High-Performance Thin-Layer Chromatography Method for the Qualitative Analysis and Quantification of Hispidulin in Hydroethanol Extract of S. barbata D. Don

Background: Hispidulin, a flavonoid renowned for its diverse pharmacological activities, is abundant in S. barbata D. Don. Given its prevalence, it is essential to establish a dependable analytical method for quantifying hispidulin in this plant. Methods: This study introduces a sensitive and reliable high-performance thin-layer chromatographic (HPTLC) method for both qualitative and quantitative analysis of hispidulin in S. barbata D. Don. Pre-coated TLC aluminum plates were utilized, and after testing various mobile phase combinations, the most effective separation was achieved with Toluene-ethyl acetate-formic acid (5:4:1, v/v/v). The method underwent extensive validation for linearity, specification, instrument precision, precision, limits of LOD and LOQ, accuracy, and robustness. Results: The developed method demonstrated excellent linearity over a specified concentration range, with a determination coefficient (r2) of 0.988. A well-defined spot at Rf 0.7, corresponding to hispidulin, was observed. The concentration of hispidulin in the hydroethanolic extract of S. barbata D. Don was quantified as 13.02 μg/mg of extract. Conclusion: In conclusion, the HPTLC method presented in this study offers a simple, accurate, and robust tool for quantitatively analyzing hispidulin in the hydroethanol extract of S. barbata D. Don. This method is suitable for routine quality control and standardization of herbal formulations containing S. barbata D. Don, thereby contributing to the advancement of research in natural product- based pharmaceuticals.