Apigenin is a naturally occurring flavone, a subclass of flavonoids characterized by a trihydroxyflavone structure with hydroxyl groups at positions 4', 5, and 7, and the molecular formula C₁₅H₁₀O₅.[1] It appears as a yellow solid with a molecular weight of 270.24 g/mol and a melting point of 345–350 °C, exhibiting solubility in ethanol, pyridine, and dilute alkalies.[1] Abundantly present in various fruits (such as cherries and apples), vegetables (including celery and parsley), herbs (like chamomile), and beverages (such as tea and beer), apigenin contributes to the nutritional profile of many common dietary plants.[2]As a bioactive compound, apigenin demonstrates potent antioxidant and anti-inflammatory properties, primarily through scavenging free radicals, regulating pro-inflammatory cytokines, and modulating signaling pathways like NF-κB and MAPK.[2] It has shown promising effects in disease prevention, including suppressing cancer progression via induction of apoptosis, cell-cycle arrest, and inhibition of angiogenesis, while exhibiting low toxicity to normal cells.[2] Additionally, apigenin supports cardiovascular health, neuroprotection, and management of conditions such as diabetes, obesity, and organ injuries in the liver, kidney, and heart by reducing oxidative stress and enhancing cellular repair mechanisms.[2]Research highlights apigenin's potential in enhancing chemotherapy efficacy and improving bioavailability through nanoformulations, addressing its moderate solubility challenges for therapeutic applications.[2] Its role in promoting sleep and mitigating aging-related effects further underscores its multifaceted pharmacological profile, positioning it as a key dietary polyphenol for health promotion.[3]
Chemistry
Structure and Properties
Apigenin is a flavone, a class of flavonoid compounds, characterized by the molecular formula C₁₅H₁₀O₅. It features a basic flavone backbone consisting of two phenyl rings (A and B) connected by a heterocyclic pyrone ring (C), with hydroxyl groups attached at the 5, 7, and 4' positions, making it 5,7,4'-trihydroxyflavone.[1]The compound has a molar mass of 270.24 g/mol and appears as a yellow crystalline solid, often isolated as yellow needles when crystallized from aqueous pyridine. It has a melting point of 345–350 °C. Apigenin exhibits negligible solubility in water (approximately 2.35 μg/mL), but it is soluble in organic solvents such as dimethyl sulfoxide (DMSO, up to 54 mg/mL) and ethanol (1 mg/mL in hot ethanol).[1][4]In terms of spectral properties, apigenin displays characteristic UV-Vis absorption maxima at 269 nm and 340 nm in ethanol, corresponding to its conjugated aromatic system, which contributes to its yellow coloration. Historically, this pigmentation has led to its use as a natural yellowdye for textiles, particularly in mordanting wool with chromium to produce fast colors resistant to soap.[1][5][6]Apigenin demonstrates basic stability under physiological conditions, remaining intact in rat plasma for up to 24 hours at room temperature and at least 4 weeks when frozen. Its reactivity includes the formation of conjugates, such as glucuronides and sulfates, through phase II metabolic processes, which enhance its solubility and facilitate excretion.[4][4]
Biosynthesis
Apigenin is synthesized in plants via the phenylpropanoid pathway, a core metabolic route for secondary metabolites that begins primarily with the amino acid L-phenylalanine, though L-tyrosine serves as an alternative precursor in certain species through bifunctional enzymes.[7][8] The pathway initiates with phenylalanine ammonia-lyase (PAL) or tyrosine ammonia-lyase (TAL), which deaminate these amino acids to yield trans-cinnamic acid or p-coumaric acid, respectively.[7] Cinnamic acid is then hydroxylated by cinnamate 4-hydroxylase (C4H) to p-coumaric acid if starting from phenylalanine.[7] This p-coumaric acid is subsequently activated by 4-coumarate:CoA ligase (4CL) to form p-coumaroyl-CoA, the key intermediate that branches into flavonoid biosynthesis.[9]From p-coumaroyl-CoA, chalcone synthase (CHS) catalyzes the condensation with three molecules of malonyl-CoA to produce naringenin chalcone, the first committed flavonoid precursor.[9]Chalcone isomerase (CHI) then cyclizes naringenin chalcone to the flavanone naringenin through stereospecific isomerization.[9] The final conversion to apigenin occurs via flavone synthase (FNS), which performs oxidative dehydration and aromatization of naringenin; in the Apiaceae family, this is mediated by the soluble FNS I, a Fe²⁺/2-oxoglutarate-dependent dioxygenase.[9][10]Biosynthesis of apigenin exhibits tissue-specific regulation, particularly in Apiaceae species like parsley (Petroselinum crispum), where FNS I expression is prominent in leaves and glandular structures to support flavone accumulation.[11] Genetic factors, including tandem gene duplications of flavanone 3-hydroxylase (F3H) leading to FNS I neofunctionalization, contribute to clustered biosynthetic genes in Apiaceae genomes, such as in celery and carrot, enhancing pathway efficiency.[9] Environmental cues like UV-B radiation and light intensity modulate pathway flux by upregulating key enzymes such as FNS and CHS, thereby increasing apigenin production as a protective response against stress.[12][13]
Occurrence
Natural Sources
Apigenin occurs in high concentrations in plants of the Apiaceae family, notably parsley (Petroselinum crispum), where dried leaves can contain up to 45 mg/g dry weight, primarily as the aglycone and glycosides.[14]Celery (Apium graveolens) also accumulates significant amounts, with concentrations reaching 0.79 mg/g in seeds and 0.24 mg/g in aerial parts like leaves and stems.[15]In the Asteraceae family, apigenin is prominent in chamomile (Matricaria chamomilla), constituting approximately 68% of total flavonoids in dried flowers, with overall contents of 3–5 mg/g dry weight.[16][17] Various Artemisia species, such as A. annua and A. vestita, contain apigenin in their aerial parts and roots, often as C- and O-glycosides, with accumulation varying by species and environmental factors.[18][19]Herbs like oregano (Origanum vulgare) accumulate up to 14 mg/g total apigenin in dried samples,[20] while thyme (Thymus vulgaris) shows lower levels around 0.04 mg/g in dried leaves.[21]Ecologically, apigenin contributes to plantdefense and adaptation, absorbing UV-B radiation to protect tissues from photodamage, as demonstrated in maize where flavone synthase enzymes produce apigenin in response to UV exposure.[12] As part of flavonoid pigments, it also aids in pollinator attraction by enhancing flower coloration and signaling in species like chamomile.[22]
Glycosides
Apigenin occurs in nature predominantly as glycosides, where sugar moieties are attached to the flavonoid core, forming O-glycosides or C-glycosides that modify its chemical properties.[4] Major glycosides include apigetrin (apigenin-7-O-glucoside), an O-linked form with a glucose attached at the 7-position of the B-ring; vitexin (apigenin-8-C-glucoside), a C-linked variant with glucose bound directly to the 8-position of the A-ring; isovitexin (apigenin-6-C-glucoside), similarly C-linked but at the 6-position; and rhoifolin (apigenin-7-O-rutinoside), featuring a disaccharide (rutinose, composed of rhamnose and glucose) at the 7-position.[4] Another notable example is apiin, a complex O-glycoside with an apiosyl-glucose unit at the 7-position.[23]These glycosides are formed in plants through the action of UDP-glycosyltransferases (UGTs), enzymes that catalyze the transfer of activated sugars, such as UDP-glucose or UDP-rhamnose, to specific hydroxyl (for O-glycosides) or carbon (for C-glycosides) positions on the apigenin aglycone, typically at sites 5, 7, or 6/8.[24] This glycosylationprocess enhances the molecule's water solubility—up to several orders of magnitude higher than the aglycone (which has a solubility of approximately 0.00135 mg/mL)—and improves chemical stability against degradation, facilitating storage and transport within plant tissues.[4][24]In terms of distribution, apigenin glycosides are widespread in various plant families, with specific prevalence in certain species; for instance, vitexin and isovitexin are abundant in bamboo leaves (e.g., up to 2.78 mg/g for related isomers), while apiin predominates in parsley seeds and leaves of Apiaceae plants like celery.[25][23] Rhoifolin occurs in citrus fruits and herbs such as oregano.[4] Analytical identification of these glycosides commonly employs high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), which separates and characterizes them based on retention times, mass-to-charge ratios, and fragmentation patterns, enabling precise quantification in plant extracts.[26]Compared to the free aglycone form, glycosylated apigenin exhibits superior bioavailability due to increased solubility, allowing better dissolution and uptake in biological systems, though the exact mechanisms vary by glycoside type.[4] In plants, these conjugates serve as storage forms of the flavonoid, acting as inactive reservoirs that can be hydrolyzed to release active aglycones during stress responses, such as UV exposure or pathogen attack, thereby contributing to defense and metabolic regulation.[27]
Dietary Aspects
Food Sources
Apigenin is most abundantly found in certain edible herbs and vegetables, with parsley serving as one of the richest sources. Dried parsley contains approximately 4503 mg of apigenin per 100 g, while fresh parsley provides about 215 mg per 100 g. Celery hearts contribute around 19 mg per 100 g, making them a notable vegetable source. In chamomile flowers, used to prepare tea, apigenin constitutes up to 68% of the total flavonoids.Dietary exposure to apigenin varies significantly based on food preparation methods. Drying concentrates apigenin levels in herbs like parsley by removing water content, whereas fresh forms retain lower concentrations due to higher moisture. For beverages such as chamomile tea, brewing time influences extraction; longer steeping (e.g., 5-10 minutes) can increase apigenin yield from 1.5 mg to 5 mg per cup, depending on flower quality and water temperature. Herbs like parsley and thyme, vegetables including celery and artichokes, and herbal teas collectively account for the majority of apigenin in typical diets.Global dietary intake estimates indicate that apigenin is a minor component of overall flavonoid consumption. In the average Western diet, daily apigenin intake ranges from 0.13 to 1.35 mg among middle-aged and older women, while in China it averages 4.23 mg per day. These levels reflect moderate contributions from everyday foods rather than high-dose sources.Several factors influence apigenin concentrations in foods. Cultivation practices, such as soil quality and sunlight exposure, can vary levels by up to 20-30% across growing regions. Seasonal differences affect biosynthesis, with higher accumulation in summer-harvested parsley and chamomile. Processing methods, including thermal treatments like boiling or drying, lead to losses of 10-50% due to degradation at elevated temperatures or pH shifts above 5.
Absorption and Metabolism
Apigenin exhibits low oral bioavailability, estimated at approximately 5-10% in humans, primarily attributed to its poor water solubility (around 2.16 μg/mL) and extensive first-pass metabolism.[28] Following dietary intake, apigenin is mainly absorbed in the small intestine, particularly the duodenum and jejunum, through passive diffusion across the intestinal epithelium, with involvement of efflux transporters such as P-glycoprotein (P-gp) and multidrug resistance-associated protein 2 (ABCC2) that can limit uptake.[29] This absorption process is saturable and occurs within 3.9 to 24 hours post-ingestion, resulting in peak plasma concentrations of about 127 nM in human studies.[28]In terms of metabolism, apigenin undergoes phase I oxidation primarily in the liver via cytochrome P450 enzymes, including CYP1A2 and CYP3A4, leading to hydroxylated metabolites such as luteolin.[28] Phase II conjugation follows, with extensive glucuronidation and sulfation by UDP-glucuronosyltransferases (UGTs) and sulfotransferases (SULTs) in the liver and intestine, producing water-soluble conjugates like apigenin glucuronides and sulfates.[29] For dietary glycosides, such as apigenin-7-glucoside, gut microbiota play a crucial role in initial deglycosylation through beta-glucosidase activity in the colon, releasing free aglycone apigenin for subsequent absorption; this microbial transformation enhances overall bioavailability compared to the intact glycoside.[30] Enterohepatic recirculation of these conjugates, via biliary secretion back into the intestine, further prolongs systemic exposure.[28]Excretion of apigenin occurs predominantly through urine as conjugated metabolites (accounting for about 51% of the dose in preclinical models) and to a lesser extent via feces (around 12%), with unabsorbed portions eliminated directly in the gut.[29] The plasma half-life of apigenin is approximately 4-8 hours in humans, reflecting rapid clearance, though terminal elimination can extend longer due to recirculation and tissue distribution.[28] Chronic dietary intake may lead to potential accumulation in tissues like the liver, as evidenced by detection in rat models up to 9 days post-dose, underscoring the need for further human studies on long-term pharmacokinetics.[29]
Biological Effects
Pharmacological Mechanisms
Apigenin exerts anxiolytic effects primarily through its interaction with the γ-aminobutyric acid type A (GABA_A) receptor, where it binds competitively to the benzodiazepine site located at the α-γ subunit interface, thereby enhancing GABA-mediated chloride influx and hyperpolarization of neurons.[31]In vitro studies using recombinant GABA_A receptors (e.g., α1β2γ2 subtypes) demonstrate that apigenin acts as a positive allosteric modulator, enhancing GABA-induced currents at concentrations around 1-10 μM.[32] This is consistent with its moderate affinity (Ki ≈ 4-30 μM) for the benzodiazepine site as measured by displacement of radiolabeled flunitrazepam in rat brain membranes.[33] Animal models, such as elevated plus-maze tests in mice, confirm this mechanism, showing dose-dependent anxiolytic activity (e.g., 0.5-5 mg/kg orally) that aligns with benzodiazepine site involvement, as effects in related flavonoid studies are reversed by the antagonistflumazenil.[31][34] Early reports indicated conflicting potency data, with some suggesting insensitivity to flumazenil and potential allosteric modulation outside the classical benzodiazepine site; however, post-2020 structural analyses of flavonoid-GABA_A interactions, including cryo-EM models of flavone-bound receptors, have clarified that apigenin's effects arise from benzodiazepine site occupancy with subtype-specific variations (e.g., stronger potentiation at α2/3-containing receptors).[35][36]In terms of anti-inflammatory actions, apigenin inhibits key enzymes in the arachidonic acid pathway, including cyclooxygenase-2 (COX-2) and lipoxygenase (LOX), thereby reducing prostaglandin and leukotriene production that drive inflammation.[2] For instance, in lipopolysaccharide (LPS)-stimulated macrophages, apigenin (10-50 μM) suppresses COX-2 expression and activity by 40-60%, as evidenced by decreased PGE2 release in vitro and in rodent models of acute inflammation (e.g., carrageenan-induced paw edema).[37] Similarly, it exhibits weak but measurable LOX inhibition (IC50 ≈ 100 μM for 5-LOX in cell-free assays), contributing to reduced leukotriene B4 in activated neutrophils. Complementing these enzymatic effects, apigenin activates the nuclear factor erythroid 2-related factor 2 (Nrf2) pathway, promoting translocation of Nrf2 to the nucleus and upregulation of antioxidant genes such as heme oxygenase-1 (HO-1) and NAD(P)H quinonedehydrogenase 1 (NQO1).[38] In oxidative stress models, like H2O2-treated human melanocytes or rat kidney cells under high-fat diet conditions, apigenin (5-20 μM) enhances Nrf2 nuclear accumulation by 2-3 fold, mitigating reactive oxygen species (ROS) and lipid peroxidation without altering Keap1 levels directly.[39]Apigenin also modulates estrogen signaling by interacting with estrogen receptors (ERα and ERβ) and inhibiting aromatase, the enzyme converting androgens to estrogens. It acts as a selective ER modulator, preferentially activating ERβ (EC50 ≈ 10 μM in reporter assays) over ERα, which promotes antiproliferative effects in ER-positive cells like breast cancer lines.[40] In vitro binding studies show apigenin displaces estradiol from ERβ with higher affinity (Ki ≈ 50 μM) than ERα, leading to recruitment of corepressors and suppression of estrogen-responsive genes in MCF-7 cells.[41] Concurrently, apigenin inhibits aromatase activity (IC50 ≈ 0.5-10 μM in microsomal assays from human placental tissue), reducing estrogen biosynthesis by up to 70% in preclinical models of hormone-dependent cancers.[42] In immune cells, such as macrophages and mast cells, apigenin interferes with nuclear factor-κB (NF-κB) signaling by inhibiting IκB kinase (IKK) phosphorylation and p65 nuclear translocation, thereby attenuating pro-inflammatory cytokine production (e.g., TNF-α, IL-6 reduced by 50-80% at 20-50 μM in LPS-challenged models).[43] Animal studies corroborate this, with oral apigenin (20 mg/kg) suppressing NF-κB activation in TRAMP mouse prostates, linking it to reduced immune-mediated inflammation.[44]
Health Applications and Safety
Apigenin has been investigated for its potential anxiolytic and sleep-promoting effects, primarily through studies on chamomile extracts containing the compound. Clinical trials involving chamomile tea or extracts, standardized to apigenin content, have demonstrated mild reductions in anxiety symptoms and improvements in sleep quality, particularly in individuals with generalized anxiety disorder or insomnia, at doses equivalent to 50-100 mg of apigenin daily.[45][46] A 2024 systematic review of randomized controlled trials confirmed these benefits, noting decreased awakenings and better sleep maintenance, though effects were modest and more pronounced in short-term use.[45]In cancer research, apigenin exhibits anti-proliferative effects in vitro by inducing apoptosis in prostate and breast cancer cells through inhibition of the PI3K/Akt pathway.[47] Limited human evidence comes from Phase I trials, such as a pilot study in high-risk breast patients testing apigenin supplementation from celery sources, which assessed safety and tolerability without reporting significant adverse events or efficacy endpoints yet.[48] Another early-phase trial explored its role in gastrointestinal cancers, highlighting potential chemopreventive actions but emphasizing the need for further validation.[47]Neuroprotective properties of apigenin have been observed in Alzheimer's disease models, where it reduces amyloid-beta deposition and inhibits tau protein hyperphosphorylation, thereby mitigating neuronal damage.[49] A 2023 study in animal models demonstrated apigenin's ability to lower amyloid-beta levels and improve cognitive function, supporting its role in attenuating neuroinflammation and oxidative stress associated with the disease.[49]For anti-inflammatory applications, apigenin shows promise in arthritis management, with animal studies indicating suppression of osteoarthritis progression via enhanced mitophagy and reduced chondrocyte pyroptosis.[50] Human observational data link higher flavonoid intake, including apigenin, to lower arthritis risk, while a clinical study using chamomile oil rich in apigenin glycosides reported significant knee pain relief in osteoarthritis patients.[51]Apigenin is generally recognized as safe (GRAS) by the FDA when consumed as part of chamomile extracts, with no major toxicity observed in chronicanimal studies up to doses of 50 mg/kg body weight.[52] Rare side effects include allergic reactions in sensitive individuals, similar to those from chamomile.[52] It may interact with CYP450 substrates, such as sedatives, by inhibiting enzymes like CYP3A4 and CYP4F2, potentially enhancing their effects or leading to toxicity; caution is advised with concurrent use.[53] Subacute toxicity studies report a no-observed-adverse-effect level (NOAEL) of 75 mg/kg for apigenin glycosides, with an LD50 exceeding 4800 mg/kg in rodents.[54]Despite these findings, evidence gaps persist, including the lack of large-scale randomized controlled trials (RCTs) to confirm clinical efficacy across applications. Recent 2024 reviews highlight advancements in nanoparticle delivery systems, such as nanoemulsions and PLGA nanoparticles, which improve apigenin's poor bioavailability and stability, potentially enabling higher therapeutic doses with reduced gastrointestinal inactivation.[55][56] As of 2025, comprehensive reviews continue to affirm apigenin's multifaceted pharmacological profile and explore emerging applications in neurodegeneration, metabolic syndrome, and bone protection.[16]