Agrimoniin

• Antioxidant Protection
• Anti-inflammatory Effects
• Antidiabetic Properties
• Cellular Defense Enhancement

• Anticancer Potential
• Antimicrobial Activity
• Cardiovascular Support
• Neuroprotection
• Gut Health
• Immune Modulation
• Metabolic Health
• Liver Protection

Agrimoniin represents a remarkable class of complex polyphenolic compounds with unique structural features and diverse biological activities. As a dimeric ellagitannin, agrimoniin possesses an intricate molecular architecture that underlies its potent biological effects across multiple physiological systems. Structurally, agrimoniin consists of two dehydrohexahydroxydiphenoyl (DHHDP)-glucose units linked through specific carbon-carbon bonds, creating a large molecule (MW ~1,870 Da) with numerous phenolic hydroxyl groups. This dimeric structure distinguishes it from monomeric ellagitannins and contributes to its enhanced biological potency.

The compound is predominantly found in plants of the Rosaceae family, particularly in Agrimonia species (agrimony), strawberries (Fragaria species), blackberries, raspberries, and other berries. As a powerful antioxidant, agrimoniin effectively neutralizes free radicals through multiple mechanisms. Its abundant phenolic hydroxyl groups readily donate hydrogen atoms to stabilize reactive oxygen species (ROS) and reactive nitrogen species (RNS) through hydrogen atom transfer (HAT) mechanisms. Additionally, agrimoniin can engage in single electron transfer (SET) processes, providing alternative pathways for radical neutralization.

The compound’s exceptional antioxidant capacity exceeds that of many smaller polyphenols due to the presence of multiple galloyl and DHHDP groups, which provide numerous sites for radical scavenging. This antioxidant activity helps protect cellular components—including lipids, proteins, and DNA—from oxidative damage, potentially slowing aging processes and preventing oxidative stress-related diseases. Beyond direct radical scavenging, agrimoniin enhances endogenous antioxidant systems by activating nuclear factor erythroid 2-related factor 2 (Nrf2), a master regulator of cellular antioxidant responses. This activation increases the expression of antioxidant enzymes such as superoxide dismutase (SOD), catalase, glutathione peroxidase (GPx), and heme oxygenase-1 (HO-1), providing a second layer of protection against oxidative stress.

The anti-inflammatory properties of agrimoniin operate through multiple mechanisms. It inhibits key inflammatory enzymes such as cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX), reducing the production of pro-inflammatory eicosanoids including prostaglandins and leukotrienes. Agrimoniin also suppresses the activation of nuclear factor-kappa B (NF-κB) and mitogen-activated protein kinases (MAPKs), key signaling pathways in inflammatory responses. This inhibition reduces the production of pro-inflammatory cytokines and mediators such as tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6).

Additionally, agrimoniin modulates the activity of inflammasomes, particularly NLRP3, which are multiprotein complexes involved in the processing and release of pro-inflammatory cytokines. The antidiabetic properties of agrimoniin are particularly noteworthy and operate through multiple complementary mechanisms. Most significantly, agrimoniin is a potent inhibitor of α-glucosidase, a key enzyme in the small intestine responsible for breaking down complex carbohydrates into absorbable monosaccharides. By inhibiting this enzyme, agrimoniin delays carbohydrate digestion and absorption, reducing postprandial glucose spikes—a critical factor in managing type 2 diabetes and metabolic syndrome.

Studies have shown that agrimoniin’s α-glucosidase inhibitory activity is comparable to or exceeds that of acarbose, a pharmaceutical α-glucosidase inhibitor used in diabetes management. Beyond enzyme inhibition, agrimoniin enhances insulin sensitivity in peripheral tissues by activating insulin receptor signaling pathways and increasing the expression and translocation of glucose transporters (particularly GLUT4) to cell membranes. This facilitates glucose uptake into muscle and adipose tissues, reducing blood glucose levels. Agrimoniin also protects pancreatic β-cells from oxidative damage and may stimulate insulin secretion under appropriate conditions.

Additionally, it inhibits protein tyrosine phosphatase 1B (PTP1B), a negative regulator of insulin signaling, further enhancing insulin sensitivity. The anticancer potential of agrimoniin involves multiple mechanisms targeting different aspects of cancer development and progression. It induces apoptosis (programmed cell death) in cancer cells through both intrinsic (mitochondrial) and extrinsic (death receptor) pathways. This involves activation of caspases, increase in the Bax/Bcl-2 ratio, and promotion of cytochrome c release from mitochondria—all key events in apoptotic cell death.

Agrimoniin inhibits cancer cell proliferation by arresting the cell cycle, typically at the G1/S or G2/M checkpoints, through modulation of cyclins and cyclin-dependent kinases (CDKs). It suppresses angiogenesis (the formation of new blood vessels) by inhibiting vascular endothelial growth factor (VEGF) and matrix metalloproteinases (MMPs), thereby limiting tumor growth and metastasis. Agrimoniin also modulates signaling pathways involved in cancer progression, including PI3K/Akt/mTOR, MAPK/ERK, and Wnt/β-catenin pathways. It has shown particular promise against breast cancer cells, including drug-resistant lines, where it can reverse multidrug resistance by inhibiting P-glycoprotein and other drug efflux pumps.

The antimicrobial activity of agrimoniin is attributed to several mechanisms. It disrupts bacterial cell membranes through interactions with membrane phospholipids and proteins, increasing membrane permeability and causing leakage of cellular contents. Agrimoniin chelates essential metal ions required for bacterial growth, such as iron, zinc, and manganese, limiting their availability to pathogens. It inhibits bacterial enzymes critical for metabolism and survival, including those involved in cell wall synthesis, protein synthesis, and DNA replication.

Additionally, agrimoniin interferes with bacterial biofilm formation by inhibiting quorum sensing systems and disrupting the extracellular polymeric substances (EPS) that form the biofilm matrix. It has demonstrated effectiveness against various pathogenic bacteria, including drug-resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA). In cardiovascular support, agrimoniin improves endothelial function by enhancing nitric oxide (NO) production through increased expression and activity of endothelial nitric oxide synthase (eNOS). It reduces LDL cholesterol oxidation, a key step in atherosclerosis development, through its potent antioxidant activities.

Agrimoniin inhibits platelet aggregation by interfering with platelet activation pathways and reducing the production of thromboxane A2. It modulates lipid metabolism by affecting the expression and activity of key enzymes involved in cholesterol and triglyceride metabolism. Additionally, agrimoniin helps maintain healthy blood pressure by enhancing NO bioavailability and inhibiting angiotensin-converting enzyme (ACE), which plays a role in blood pressure regulation. The neuroprotective effects of agrimoniin stem from multiple mechanisms.

It reduces oxidative stress and inflammation in neural tissues, protecting neurons from damage. Agrimoniin modulates neurotransmitter systems, including cholinergic, dopaminergic, and GABAergic pathways, influencing cognitive function, mood, and neuroprotection. It may also inhibit protein aggregation associated with neurodegenerative diseases, such as amyloid-beta in Alzheimer’s disease and alpha-synuclein in Parkinson’s disease. For gut health, agrimoniin acts as a prebiotic, promoting the growth of beneficial gut bacteria such as Lactobacillus and Bifidobacterium species while inhibiting pathogenic species.

It strengthens intestinal barrier function by enhancing tight junction proteins and reducing intestinal permeability. Agrimoniin reduces gut inflammation through the mechanisms described earlier and modulates gut-brain axis signaling, potentially influencing systemic health beyond the digestive system. In terms of immune modulation, agrimoniin enhances innate immune responses by activating macrophages and natural killer (NK) cells, increasing phagocytosis and cytotoxic activity. It modulates adaptive immunity by influencing T cell differentiation, typically promoting anti-inflammatory Th2 and regulatory T cell responses while suppressing pro-inflammatory Th1 and Th17 responses.

Agrimoniin also enhances antibody production by B cells and modulates cytokine and chemokine networks to balance immune responses. For liver protection, agrimoniin reduces oxidative stress in hepatocytes and inhibits lipid peroxidation, protecting liver cells from damage. It enhances the activity of liver detoxification enzymes, particularly phase II enzymes involved in conjugation reactions. Agrimoniin also reduces hepatic inflammation and may inhibit hepatic stellate cell activation, potentially reducing liver fibrosis.

The bioavailability of intact agrimoniin is limited due to its large molecular size and complex structure. However, it undergoes extensive metabolism in the gut, particularly by intestinal microbiota, producing smaller, more absorbable compounds that contribute to its systemic effects. The primary metabolites include ellagic acid (formed by hydrolysis of the ellagitannin structure) and subsequently, urolithins (particularly urolithin A, B, C, and D) produced by gut bacterial metabolism of ellagic acid. These metabolites can be absorbed into the bloodstream and reach various tissues, exerting biological activities that may differ from but complement those of the parent compound.

This metabolic transformation represents a crucial aspect of agrimoniin’s mechanism of action, as many of its systemic effects may be mediated by these metabolites rather than the intact molecule.

Establishing precise dosage recommendations for agrimoniin presents unique challenges due to several factors. First, agrimoniin is typically consumed as a component of complex plant extracts or whole foods rather than as an isolated compound. Second, the concentration of agrimoniin in these sources varies widely depending on factors such as plant species, growing conditions, harvest time, and extraction methods. Third, limited clinical studies have been conducted specifically on agrimoniin as an isolated compound, making evidence-based dosing recommendations difficult.

Finally, individual variations in metabolism, gut microbiota composition, and health status can significantly influence the bioavailability and effects of agrimoniin and its metabolites. Despite these challenges, the following dosage information is derived from the available research on agrimoniin-containing plants and extracts, as well as traditional usage patterns.

With Meals: For general antioxidant and anti-inflammatory effects, consumption with meals may enhance absorption through food matrix effects and reduce potential gastric irritation

Before Meals: For antidiabetic effects, consumption 15-30 minutes before carbohydrate-containing meals maximizes α-glucosidase inhibition and postprandial glucose management

Consistency: Regular, daily intake appears more beneficial than occasional high doses, based on traditional usage patterns and the cumulative nature of many effects

Duration Of Use: Short-term use (1-2 weeks) may provide some acute benefits, but many effects (particularly anti-inflammatory and metabolic) develop more fully with consistent use over 4-12 weeks

Current dosage recommendations are primarily extrapolated from traditional usage, in vitro studies, and limited animal research. Clinical trials with standardized agrimoniin preparations are lacking, and optimal therapeutic doses for specific health conditions have not been established. Individual variations in metabolism, gut microbiota, and response further complicate dosing recommendations. Future research should focus on dose-response relationships, bioavailability from different sources, and clinical outcomes with standardized preparations.

Agrimoniin presents significant bioavailability challenges due to its large molecular size (approximately 1,870 Da), complex dimeric structure, and high degree of hydroxylation.

These physicochemical properties severely limit passive diffusion across intestinal membranes. The bioavailability of intact agrimoniin is estimated to be extremely low (<0.1%), with negligible amounts of the parent compound detected in systemic circulation after oral consumption.

However ,

this limited direct absorption does not diminish agrimoniin’s biological significance, as

it undergoes extensive metabolism by gut microbiota and exerts local effects in the gastrointestinal tract before its metabolites enter systemic circulation.

Gut Microbiota Metabolism: The most significant route of agrimoniin biotransformation involves gut microbiota. The first step typically involves hydrolysis of the ester bonds in the ellagitannin structure, releasing ellagic acid. This hydrolysis can occur through both chemical processes in the acidic environment of the stomach and enzymatic processes mediated by intestinal and microbial esterases. Subsequently, ellagic acid undergoes further metabolism by specific gut bacteria, particularly Gordonibacter, Lactobacillus, and Bifidobacterium species, which perform decarboxylation, dehydroxylation, and reduction reactions to produce urolithins (particularly urolithin A, B, C, and D). These transformations occur sequentially, with urolithin D typically formed first, followed by urolithin C, urolithin A, and finally urolithin B, though the specific pattern varies between individuals based on their gut microbiota composition.

Phase I Metabolism: Hepatic cytochrome P450 enzymes play a minor role in agrimoniin metabolism, primarily affecting any absorbed ellagic acid or intermediate metabolites. CYP1A2 may be involved in hydroxylation reactions, though this represents a secondary metabolic pathway compared to gut microbial metabolism.

Phase II Metabolism: Conjugation reactions, particularly glucuronidation, sulfation, and methylation, significantly affect the metabolites of agrimoniin, especially urolithins and any absorbed ellagic acid. These reactions are catalyzed by UDP-glucuronosyltransferases (UGTs), sulfotransferases (SULTs), and catechol-O-methyltransferases (COMT), respectively. The resulting conjugates typically exhibit increased water solubility and altered biological activity compared to their precursors. Urolithin A and B glucuronides represent the predominant metabolites detected in human plasma after consumption of agrimoniin-rich foods.

Enterohepatic Circulation: Some agrimoniin metabolites, particularly glucuronide conjugates of urolithins, undergo enterohepatic circulation. These conjugates can be excreted in bile, deconjugated by intestinal β-glucuronidases, and reabsorbed, prolonging their presence in the body and potentially extending their biological effects.

Plasma Protein Binding: Urolithins and ellagic acid, the primary circulating metabolites derived from agrimoniin, exhibit moderate to high plasma protein binding (60-95%), primarily to albumin. This binding affects their free concentration and distribution to tissues.

Tissue Distribution: Distribution studies with agrimoniin are limited, but research with urolithins suggests that these metabolites may accumulate in various tissues including prostate, intestinal tissues, and colon. Urolithins have been detected in prostate tissue after consumption of ellagitannin-rich foods, suggesting potential relevance for prostate health. Limited evidence suggests some distribution to liver, kidney, and adipose tissue, with minimal penetration of the blood-brain barrier.

Blood-brain Barrier Penetration: Intact agrimoniin does not cross the blood-brain barrier due to its size and polarity. Some urolithin metabolites, particularly those that undergo phase II metabolism to increase lipophilicity, may achieve limited CNS penetration, though this remains poorly characterized.

Primary Routes: Renal excretion represents the primary elimination route for agrimoniin metabolites, particularly for conjugated urolithins. Fecal elimination is significant for unabsorbed parent compounds, unmetabolized ellagic acid, and metabolites excreted via bile.

Half-life: The elimination half-life varies significantly among different metabolites: urolithins typically exhibit half-lives of 12-48 hours, with conjugated forms generally having longer half-lives than free forms. Ellagic acid has a relatively short plasma half-life of 8-14 hours.

Clearance Factors: Renal function significantly impacts the clearance of agrimoniin metabolites. Age, hydration status, and concurrent medications affecting renal function may therefore influence their elimination kinetics.

Plasma Biomarkers: Urolithins (particularly urolithin A, B, and their glucuronide conjugates) serve as the primary biomarkers of agrimoniin consumption and metabolism. These can be detected in plasma typically 12-48 hours after consumption, with peak levels often observed at 24-72 hours, reflecting the time required for microbial metabolism.

Urinary Biomarkers: Urinary excretion of urolithin glucuronides and sulfates provides a non-invasive measure of agrimoniin metabolism. These metabolites can be detected in urine for up to 96 hours after consumption, with a characteristic pattern that reflects individual metabotype.

Analytical Methods: Liquid chromatography-mass spectrometry (LC-MS/MS) represents the gold standard for detecting and quantifying agrimoniin metabolites in biological samples. High-performance liquid chromatography (HPLC) with electrochemical or fluorescence detection may also be used for specific metabolites.

Gastrointestinal Effects: Many of agrimoniin’s biological effects may occur locally in the gastrointestinal tract before any absorption. These include direct antioxidant activity in the gut lumen, antimicrobial effects against pathogenic bacteria, prebiotic effects promoting beneficial gut microbiota, α-glucosidase inhibition affecting carbohydrate digestion, and direct anti-inflammatory effects on intestinal epithelium.

Systemic Effects: Systemic effects are primarily mediated by absorbed metabolites, particularly urolithins and their conjugates. These include anti-inflammatory activities in various tissues, modulation of cellular signaling pathways, and potential effects on energy metabolism and mitochondrial function.

Relative Importance: Both local and systemic effects contribute to agrimoniin’s overall health benefits, with the relative importance varying by specific health outcome. For gut health, antimicrobial effects, and glycemic response, local effects may predominate, while for cardiovascular, anti-inflammatory, and some anticancer effects, systemic metabolites likely play the major role.

Significant knowledge gaps remain regarding agrimoniin bioavailability.

These include limited understanding of the specific transporters involved in metabolite absorption, incomplete characterization of the microbial enzymes responsible for urolithin production, limited data on tissue distribution of metabolites, and insufficient clinical studies correlating metabolite levels with biological effects.

Additionally , the impact of various formulation approaches on bioavailability requires further investigation through controlled clinical trials. The relationship between metabotype, urolithin production, and health outcomes represents a particularly important area for future research.

Agrimoniin generally demonstrates a favorable safety profile based on its natural occurrence in commonly consumed foods and medicinal plants with long histories of traditional use. The safety rating of 4 out of 5 reflects the good historical safety record of agrimoniin-containing plants, limited reports of adverse effects, and the absence of significant toxicity in available studies.

However ,

this rating acknowledges that comprehensive toxicological evaluations of isolated agrimoniin are limited, and certain populations may experience adverse reactions or interactions. Most safety data is derived from the consumption of agrimoniin-containing plants and foods rather than isolated compounds, and the safety profile of concentrated extracts or supplements may differ from traditional dietary sources.

Common Mild:

Rare Serious:

Established Ul: No officially established upper limit by regulatory authorities

Research Based Limit: Limited toxicological data on isolated agrimoniin; safety studies with agrimony extracts suggest no adverse effects at doses providing up to 200-300 mg agrimoniin daily

Practical Recommendation: For concentrated extracts, doses providing more than 200 mg agrimoniin daily are not recommended without medical supervision. For food sources and traditional preparations, consumption within historical usage patterns is likely safe.

Acute Toxicity:

• Not established for isolated agrimoniin; agrimony extracts show very low acute toxicity with LD50 >5000 mg/kg in rodent studies
• Limited acute toxicity expected based on historical consumption patterns and preliminary animal studies
• No specific target organ toxicity identified at typical exposure levels

Chronic Toxicity:

• Limited formal long-term toxicity studies; 90-day rodent studies with agrimony extracts show no significant adverse effects at doses equivalent to 5-10 times traditional human consumption
• Estimated at 100-200 mg/kg/day agrimoniin equivalent in rodent studies
• Centuries of human consumption of agrimoniin-containing plants suggests low chronic toxicity at traditional intake levels

Genotoxicity:

• Negative for mutagenicity in bacterial reverse mutation assays with agrimony extracts
• No significant clastogenic activity observed in limited studies with agrimony extracts
• Negative in in vivo micronucleus tests with agrimony extracts

Carcinogenicity:

• No dedicated carcinogenicity studies with agrimoniin or containing plants
• No evidence suggesting carcinogenic potential; some epidemiological data suggests potential protective effects against certain cancers

Reproductive Toxicity:

• Limited data; traditional use of agrimony has not been associated with fertility concerns
• Insufficient data specific to agrimoniin; traditional use of moderate amounts of agrimony not associated with birth defects
• Avoid concentrated sources during pregnancy and lactation due to limited safety data; food sources likely safe

Pediatric: Food sources (berries) appropriate; medicinal doses of extracts should be adjusted by weight and limited to children over 6 years

Geriatric: Generally well-tolerated; consider reduced dosing due to potential changes in metabolism and elimination; monitor for drug interactions

Renal Impairment: No specific contraindications, but consider reduced dosing in severe impairment due to altered metabolism of polyphenols

Hepatic Impairment: Use with caution in significant liver disease; metabolic burden may be increased

Genetic Considerations: Individuals with specific polymorphisms affecting polyphenol metabolism may experience altered effects or tolerability; metabotype variations affect urolithin production

Known Allergens: Rare allergic reactions to plants in the Rosaceae family have been reported; cross-reactivity possible

Cross Reactivity: Potential cross-reactivity with other Rosaceae plants (strawberries, apples, almonds, etc.)

Testing Methods: No standardized testing available for agrimoniin sensitivity; conventional allergy testing for Rosaceae plants may be relevant

Management: Discontinue use if allergic symptoms develop; consider allergy consultation for severe reactions

Recommended Monitoring: No specific monitoring required for typical consumption of food sources or traditional preparations

Parameters Of Concern: For concentrated sources or therapeutic doses: blood glucose (in diabetics), bleeding parameters (in those on anticoagulants), liver function (in those with hepatic concerns)

Frequency: Baseline and periodic monitoring may be considered for long-term use of concentrated forms

Symptoms: Primarily gastrointestinal distress, including nausea, vomiting, abdominal pain, and diarrhea. Severe astringency in mouth and throat. Potential hypoglycemia in susceptible individuals.

Management: Supportive care; activated charcoal may be considered for recent significant ingestion of concentrated forms; monitor blood glucose in symptomatic cases

Antidote: No specific antidote; treatment is symptomatic and supportive

Agrimoniin in its natural sources (berries, agrimony, other Rosaceae plants) has an established safety record when consumed in traditional amounts. The matrix effects of these foods, including the presence of other compounds, contribute to their overall safety profile. The safety considerations for isolated or concentrated agrimoniin may differ from these traditional sources.

Pharmacokinetic Interactions: May affect absorption of certain minerals (particularly iron and zinc) and potentially some drugs through formation of insoluble complexes. Limited evidence for significant effects on drug metabolism enzymes at typical doses.

Pharmacodynamic Interactions: Most significant interactions are pharmacodynamic, particularly additive effects with antidiabetic, antiplatelet, and antihypertensive medications.

Monitoring Recommendations: Most important to monitor blood glucose when used with antidiabetic medications and bleeding parameters when used with anticoagulants.

Historical Contraindications: Traditional herbal texts often caution against high doses of agrimony in pregnancy and in those with ‘hot and dry’ constitutions (likely referring to individuals prone to constipation or gastric irritation).

Traditional Preparation Methods: Traditional preparation methods often include combining agrimony with demulcent herbs or honey, which may mitigate potential gastric irritation from tannins.

Historical Dosing Limits: Traditional European herbal practice typically limited agrimony tea to 2-3 cups daily, which aligns with modern safety assessments.

Agrimoniin currently has no specific regulatory status as an isolated compound in most jurisdictions worldwide. It is primarily regulated indirectly as a naturally occurring component of herbs and foods that have been part of human diets for centuries. Agrimony (Agrimonia eupatoria and related species), strawberries, and other berries containing agrimoniin are generally recognized as safe for consumption based on their long history of use. As research into agrimoniin’s biological activities progresses, its regulatory status may evolve, particularly if isolated forms are developed for supplement or pharmaceutical applications.

Currently, no major regulatory agency has issued specific guidance or restrictions on agrimoniin beyond those applying to its traditional sources.

Fda Status: No specific regulatory classification for isolated agrimoniin. As a component naturally present in foods and herbs with a history of safe use, it falls under existing regulations for these products.

Dietary Supplement Status: Agrimony is listed in the FDA’s ‘Old Dietary Ingredient’ list, allowing its use in dietary supplements without New Dietary Ingredient notification. Isolated agrimoniin would likely require NDI notification if marketed as a dietary supplement ingredient without a history of use in that form.

Gras Status: Strawberries, blackberries, and other common food sources of agrimoniin are Generally Recognized as Safe (GRAS) based on their history of consumption. Agrimony does not have formal GRAS status but has a history of use in teas and herbal preparations.

Health Claim Eligibility: No approved health claims specifically mentioning agrimoniin. Structure/function claims for agrimoniin-containing supplements would be permitted with appropriate disclaimers under DSHEA regulations, but disease claims would not be allowed without drug approval.

Novel Food Status: Agrimony and common berries containing agrimoniin are not considered novel foods as they have a history of consumption in the EU before May 15, 1997. Isolated agrimoniin would likely be considered a novel food ingredient if marketed for food use outside its traditional context.

Herbal Medicine Status: Agrimony has an established European Medicines Agency (EMA) monograph supporting its registration as a Traditional Herbal Medicinal Product for minor digestive disorders and minor skin inflammations. This provides a simplified registration pathway in EU member states., Not currently approved under the ‘well-established use’ pathway, which would require more substantial clinical evidence.

Food Supplement Regulations: Agrimony is permitted in food supplements across most EU member states. National variations exist in maximum levels and specific requirements. Isolated agrimoniin is not specifically addressed in food supplement regulations.

Health Claims: No authorized health claims under the EU Nutrition and Health Claims Regulation for agrimoniin or agrimony. Any claims would require EFSA scientific assessment and approval.

Investigational Status: No major pharmaceutical development programs specifically targeting isolated agrimoniin have reached advanced clinical stages. Any pharmaceutical development would require standard IND/NDA pathways in the US or equivalent processes in other jurisdictions.

Botanical Drug Pathways: The FDA’s Botanical Drug Development pathway could potentially apply to highly standardized agrimoniin-rich extracts, though this would still require substantial clinical evidence of safety and efficacy.

Orphan Drug Potential: Unlikely to qualify for orphan drug designation as potential applications (diabetes, inflammation) affect large populations exceeding orphan thresholds.

Intellectual Property Considerations: Limited patent protection potential for the naturally occurring compound itself, though novel formulations, synthetic derivatives, or specific therapeutic applications might be patentable.

Increasing Standardization: Growing regulatory emphasis on standardization and quality control for botanical products, potentially leading to more specific requirements for agrimoniin content in standardized extracts.

Evidence Requirements: Trend toward requiring stronger evidence for health claims, particularly in the EU and Australia, affecting marketing of agrimoniin-containing products.

Traditional Use Recognition: Continuing development of regulatory pathways that recognize traditional use evidence while ensuring safety and quality, potentially benefiting agrimony products with long historical use.

Global Harmonization: Efforts toward international harmonization of botanical regulations through organizations like the International Regulatory Cooperation for Herbal Medicines (IRCH), potentially creating more consistent approaches to agrimoniin-containing products.

Agrimoniin currently exists primarily as a component of plant extracts and whole foods rather than as an isolated commercial product. Its market value is therefore largely embedded within the pricing of herbal products, particularly agrimony extracts, and to a lesser extent, berry-based products where it is one of many bioactive compounds. The market for isolated agrimoniin is limited to research applications, with small quantities of analytical standards available at high prices. No significant commercial market exists for agrimoniin supplements or pharmaceutical products, though this may change as research advances.

The cost-efficiency analysis must therefore consider both the traditional sources and potential future isolated applications.

Current Market Size: The current market for agrimoniin-containing products is relatively small, estimated at $50-100 million globally, primarily in traditional herbal products and some specialized supplements.

Growth Projections: Growing interest in natural alpha-glucosidase inhibitors for diabetes management and increasing research on ellagitannins for various health applications suggest potential for 15-25% annual growth in this segment over the next 5-10 years.

Market Drivers: Rising prevalence of diabetes and prediabetes, growing consumer preference for natural health products, and increasing scientific validation of traditional remedies are key factors driving market potential.

Barriers To Growth: Limited consumer awareness, variable product quality and standardization, and competition from established pharmaceutical options represent significant barriers to market expansion.

Agrimoniin exhibits moderate to poor stability as an isolated compound due to its complex dimeric structure with numerous phenolic hydroxyl groups susceptible to oxidation and hydrolysis. Its stability is highly context-dependent, with significantly greater stability observed in its natural plant matrices compared to purified forms. The compound is particularly sensitive to alkaline conditions, oxidation, elevated temperatures, and certain metal ions. Understanding

these stability factors is crucial for preserving agrimoniin content in both natural sources and any potential isolated preparations.

In Natural Sources: In properly stored dried plant materials such as agrimony herb, agrimoniin demonstrates moderate stability, with significant content persisting for 18-24 months under optimal storage conditions (cool, dry, protected from light). In fresh or frozen berries, stability is highly dependent on storage conditions, with frozen berries (-18°C or below) retaining 70-90% of initial agrimoniin content for 6-12 months. Freeze-dried berries show better stability than fresh or conventionally frozen products.

As Isolated Compounds: Isolated agrimoniin has very limited shelf life, typically 1-3 months when stored as dry powder under inert gas at -20°C. At room temperature, shelf life decreases to 1-2 weeks even with protective packaging. In solution, stability is highly dependent on solvent, pH, and storage conditions, ranging from hours (aqueous solutions at neutral pH) to days (acidified alcoholic solutions stored cold and protected from light).

In Standardized Extracts: Standardized extracts containing agrimoniin alongside other polyphenols typically show intermediate stability, with shelf lives of 12-24 months when properly formulated with stabilizers and stored under appropriate conditions. Dry extracts generally exhibit better stability than liquid formulations, and hydroalcoholic extracts (tinctures) show better stability than purely aqueous preparations.

Temperature: For isolated compounds: -20°C for long-term storage of dry powders; 2-8°C for extracts and formulations intended for use within weeks. For dried herbs: cool, dry conditions (15-25°C) in sealed containers. For berries: frozen storage at -18°C or below for maximum retention of agrimoniin.

Light: Store all forms protected from light, particularly UV and blue wavelengths. Amber or opaque containers are recommended. For commercial products, packaging should provide adequate light protection.

Humidity: For dry powders and extracts, low humidity (<40% RH) prevents moisture absorption that can accelerate degradation. For dried herbs, relative humidity should be maintained below 60% to prevent moisture absorption and potential microbial growth.

Packaging: Oxygen-impermeable packaging with minimal headspace is ideal for all forms. For isolated compounds, storage under inert gas (nitrogen or argon) in sealed containers with moisture-proof barriers provides optimal protection. For commercial products, laminated foil pouches or blister packs often provide better protection than plastic bottles.

Handling: Minimize exposure to air during handling. For analytical work, process samples quickly and keep cold when possible. Avoid metal spatulas or containers when handling purified agrimoniin or concentrated extracts.

Dried Herbs: Agrimoniin shows good stability in properly dried and stored herbs such as agrimony, with retention of 70-80% of initial content after 18-24 months under optimal conditions. The natural plant matrix, including other polyphenols, fibers, and cellular structures, provides some protection against degradation. Traditional air-drying at moderate temperatures (30-40°C) generally preserves agrimoniin better than high-temperature drying methods.

Fresh Berries: In fresh berries, agrimoniin stability is limited, with significant losses occurring within days at room temperature and 1-2 weeks under refrigeration (2-8°C). Factors affecting stability include endogenous enzymes, microbial growth, and physical damage that disrupts cellular compartmentalization and exposes agrimoniin to degradative enzymes and oxygen.

Frozen Berries: Freezing significantly extends agrimoniin stability in berries, with retention of 70-90% of initial content for 6-12 months at -18°C or below. Rapid freezing (blast freezing) generally preserves more agrimoniin than slow freezing methods by limiting ice crystal formation and cellular damage.

Aqueous Extracts: Aqueous extracts (teas, infusions) show poor stability, with significant degradation occurring within hours at room temperature or days under refrigeration. Acidification (pH 3-4) and addition of antioxidants significantly improve stability.

Hydroalcoholic Extracts: Tinctures and other hydroalcoholic extracts (typically 25-60% ethanol) show intermediate stability, with retention of 60-80% of initial agrimoniin content for 12-24 months under appropriate storage conditions. The alcohol content inhibits microbial growth and provides some protection against enzymatic degradation.

Solid Dosage Forms: Tablets and capsules containing agrimoniin-rich extracts show variable stability depending on formulation. Excipients, processing conditions, and packaging all significantly impact stability. Enteric-coated formulations may provide better stability by protecting contents from oxygen and moisture until release in the intestine.

Drying: Traditional air-drying of herbs at moderate temperatures (30-40°C) typically preserves 70-90% of initial agrimoniin content. Higher temperature drying methods (>50°C) can cause significant losses, while freeze-drying generally provides the best retention (90-95%) but at higher cost.

Extraction: Extraction temperature significantly impacts agrimoniin stability, with cold or room temperature extraction preserving more compound than hot extraction methods. However, extraction efficiency must be balanced against stability considerations, as higher temperatures increase extraction yield but may decrease stability.

Concentration: Evaporation of solvents to produce concentrated extracts can cause significant agrimoniin degradation, particularly if high temperatures are used. Vacuum concentration at temperatures below 40°C helps preserve content, while spray drying should use inlet temperatures below 120°C and outlet temperatures below 80°C to minimize degradation.

Pasteurization: Thermal pasteurization of berry products typically causes 20-40% loss of agrimoniin content, with the extent depending on specific time-temperature combinations. High-temperature short-time (HTST) methods generally preserve more content than lower-temperature extended-time methods.

Cooking: Cooking of agrimoniin-containing foods causes variable losses depending on method, temperature, and duration. Boiling typically causes greater losses (40-70%) than steaming or microwave cooking (20-40%) due to both thermal degradation and leaching into cooking water.

Sample Preparation: For accurate analysis, samples containing agrimoniin should be processed quickly, kept cold, and protected from light and air. Addition of antioxidants (ascorbic acid, 0.1-1.0%) and acidification (pH 3-4) during sample preparation significantly improves stability. For plant materials, freeze-drying before extraction is preferable to air-drying for analytical purposes.

Chromatographic Analysis: During HPLC or UPLC analysis, acidified mobile phases (0.1-1.0% formic or acetic acid) improve stability during separation. Column temperature should be kept moderate (20-30°C) to minimize on-column degradation. For mass spectrometry detection, negative ionization mode typically provides better sensitivity and less fragmentation of agrimoniin.

Storage Of Standards: Analytical standards should be stored as dry powders at -80°C for long-term stability or -20°C for routine use. Working solutions should be prepared fresh or stored at -20°C in acidified (pH 3-4) 50% methanol or ethanol for no more than 1 month, with aliquoting to avoid repeated freeze-thaw cycles.

Accelerated Stability: Accelerated conditions (elevated temperature, light exposure, oxidative stress) can predict long-term stability through Arrhenius kinetics and other models. Typical conditions include 40°C/75% RH, with sampling at regular intervals for up to 6 months.

Real Time Stability: Storage under recommended conditions with periodic testing provides the most reliable stability data but requires longer timeframes (1-3+ years).

Analytical Methods: HPLC-UV or HPLC-MS are the primary methods for monitoring agrimoniin stability, with detection at 280 nm (UV) or specific mass transitions (MS). Functional assays, particularly α-glucosidase inhibition, provide valuable complementary data on biological activity retention.

Stress Testing: Deliberate exposure to extreme conditions (high temperature, strong oxidants, extreme pH) helps identify degradation pathways and products, informing stabilization strategies and analytical method development.

Historical Preservation Methods: Traditional herbal knowledge includes effective preservation techniques developed through centuries of observation. These include proper harvest timing (often during flowering for maximum content), careful drying in shade or gentle warmth, storage in sealed ceramic or glass containers, and addition of natural preservatives like alcohol in tinctures.

Seasonal Considerations: Traditional herbalists recognized seasonal variations in plant potency and developed harvest calendars to optimize active compound content. For agrimony, harvest during flowering (typically mid-summer) maximizes agrimoniin content.

Compatibility Knowledge: Traditional medicine systems developed empirical understanding of compatible and incompatible combinations. For agrimoniin-rich herbs, traditional formulations often include complementary herbs that enhance stability or efficacy, such as mild acids (hibiscus, rose hips) that create optimal pH conditions.

Natural Sources

Extraction Methods

Purification Methods

Quality Considerations

Commercial Production

Sustainable Sourcing Practices

Geographical Considerations

Identification And Authentication

Initial Discovery: While agrimoniin-containing plants have been used medicinally for millennia, agrimoniin itself was not isolated and characterized until the late 1970s and early 1980s by Japanese researchers led by Takuo Okuda. The compound was first isolated from Agrimonia pilosa (Chinese agrimony) and subsequently identified in other Rosaceae family plants. The complex dimeric structure was elucidated through a combination of chemical degradation studies, spectroscopic methods, and X-ray crystallography.

Naming Origin: The name ‘agrimoniin’ derives directly from the genus Agrimonia, from which it was first isolated. The genus name itself has ancient origins, possibly from the Greek ‘argemone’ (a plant used to treat cataracts) or ‘agremone’ (a plant used for eye ailments).

Structural Elucidation: The complete structural elucidation of agrimoniin was a significant achievement in natural product chemistry due to the compound’s complex dimeric nature. Initial studies established it as a dimeric ellagitannin composed of two dehydrohexahydroxydiphenoyl (DHHDP)-glucose units linked through specific carbon-carbon bonds. Subsequent research refined this understanding, identifying the precise stereochemistry and confirming its status as a bis-DHHDP-glucose dimer with a molecular weight of approximately 1,870 Da.

Agrimoniin has been the subject of increasing scientific research over the past two decades, with evidence supporting its efficacy for various applications, particularly in antidiabetic, antioxidant, and anti-inflammatory contexts. The strongest evidence exists for its α-glucosidase inhibitory activity and antioxidant properties, with growing support for antimicrobial, anticancer, and cardiovascular applications. In vitro and animal studies consistently demonstrate agrimoniin’s biological activities, with emerging but still limited human clinical trials. The evidence rating of 3 out of 5 reflects moderate support from scientific research, with good mechanistic understanding but limitations in the size, design, and consistency of human studies.

Much of the clinical evidence comes from studies using agrimoniin-containing plant extracts rather than the isolated compound, making it challenging to attribute effects specifically to agrimoniin versus other bioactive components.

Limited long-term clinical trials (>1 year) evaluating safety and efficacy, Insufficient dose-response studies to establish optimal therapeutic dosages, Limited research on agrimoniin-microbiome interactions and their impact on health outcomes, Incomplete understanding of the relationship between urolithin metabotypes and clinical response, Few studies directly comparing agrimoniin to established pharmaceutical interventions, Limited research on potential applications beyond diabetes, particularly in neurological conditions, Insufficient standardization of extracts and analytical methods across studies, Limited investigation of potential synergistic effects with other bioactive compounds