Safety Controversies and Development Trends of Butylated Hydroxyanisole

1 Introduction

Lipid oxidation is a primary cause of oil rancidity, nutrient loss in foods, and ageing of polymer materials. Adding antioxidants is the most cost-effective approach to slow oxidative deterioration and reduce industrial losses. Butylated hydroxyanisole (BHA, EU code E320) is a synthetic phenolic antioxidant with a long history of industrial use. It consists mainly of two isomers: 3-BHA and 2-BHA. The 3-BHA isomer accounts for more than 90% of commercial BHA. It is the principal contributor to antioxidant activity, but also the main source of biological toxicity [1,2]. BHA possesses strong thermal stability, high antioxidant efficiency, broad system compatibility, and favorable cost-effectiveness. These advantages have led to its long-term application in food processing, feed production, plastics, rubber, and cosmetics [3,4,5].

Over the past decade, progress in molecular toxicology, metabolomics, network toxicology, and environmental trace detection technologies has prompted major authorities_including the International Agency for Research on Cancer (IARC), the European Food Safety Authority (EFSA), and the U.S. National Toxicology Program (NTP)–to repeatedly update their safety evaluations of BHA. The traditional view that “compliant use equals safety” is no longer tenable [6,7]. Dietary intake is the main route of human exposure. Both BHA and its major metabolite, tert-butylhydroquinone (TBHQ), have been detected in environmental waters, sediments, and human biospecimens. Long-term exposure can cause multi-organ, multi-target toxicity, posing potential threats to ecosystems and human health [2]. In February 2026, the U.S. Food and Drug Administration (FDA) initiated a comprehensive post-market safety reassessment of BHA. This is the first systematic risk review since BHA was classified as Generally Recognized As Safe (GRAS) in 1958. It signals a shift towards more rigorous and refined global regulation [8].

Currently, research on BHA has evolved from performance evaluation to mechanistic toxicology, exposure assessment, toxicity-reduction technologies, and the development of green alternatives. Based on key Chinese and English publications and authoritative reports from 2016 to 2026, this paper systematically summarizes the fundamental properties, antioxidant mechanisms, metabolic pathways, and environmental behavior of BHA. It analyses the multi-dimensional toxicological effects and safety debates, compares global regulatory frameworks, highlights current research frontiers, and projects future trends. This review is intended to support safer application, improved risk management, and the design of next-generation antioxidants.

2 Physicochemical Properties and Structural Characteristics of BHA

The chemical name of BHA is butylated hydroxyanisole (CAS No. 25013-16-5). Its molecular formula is C₁₁H₁₆O₂, and its relative molecular mass is 180.25. At ambient temperature and pressure, BHA appears as a white to pale yellow crystalline powder with no pungent odour. It has stable physicochemical properties. Its melting point ranges from 69.5 to 71.5 °C, and its boiling point is 265 °C. BHA maintains structural integrity during high-temperature processing up to 180 °C. At 200 °C, the decomposition rate is only about 30%. This makes BHA suitable for various thermal food processes, including frying, baking, and oil refining.

BHA is highly lipophilic. It has low solubility in water and glycerol, but dissolves readily in organic solvents such as ethanol, ether, and vegetable or animal oils. This property underpins its widespread use in lipid-rich foods and hydrophobic industrial materials [3]. Commercial BHA is predominantly 3-tert-butyl-4-hydroxyanisole (3-BHA). The isomeric structure significantly influences both antioxidant potency and biological toxicity. Recent toxicological evidence indicates that 3-BHA (the ortho-isomer) exerts obesogenic and endocrine-disrupting effects in vivo, whereas 2-BHA (the meta-isomer) does not show obvious toxicity [9].

3 Antioxidant Mechanisms of BHA

3.1 Core Mechanism of Free–Radical Scavenging

The primary antioxidant action of BHA is to interrupt the chain reaction of lipid oxidation. Its phenolic hydroxyl group donates hydrogen atoms to highly reactive species_such as lipid radicals and peroxyl radicals_generated during oxidation. This converts unstable radicals into stable molecules, thereby terminating the oxidative chain amplification. Consequently, oil rancidity and material ageing are inhibited [8]. The radical species formed after BHA oxidation are stabilized by the conjugated benzene ring system, which prevents secondary oxidation and ensures sustained protective effects. In vitro studies have confirmed that BHA possesses strong radical-scavenging and chain-breaking capacities. However, its apparent activity in DPPH, ABTS, and similar assays is significantly influenced by concentration, reaction medium, and measurement conditions [10].

3.2 Regulation of Oxidative Stress Pathways

In addition to direct radical scavenging, BHA exerts indirect antioxidant effects by modulating systemic oxidative stress signaling. It can activate the Nrf2 antioxidant pathway, thereby inducing the synthesis of endogenous detoxification and antioxidant enzymes_such as glutathione S-transferase and glucuronosyltransferase_in key metabolic organs, including the liver and intestine. This enhances the body’s intrinsic antioxidant defence and alleviates basal oxidative damage [10]. Importantly, this regulatory effect is dose-dependent and bidirectional. At low doses, BHA predominantly activates protective responses. At higher doses or under continuous exposure, BHA and its reactive metabolites may also exert pro-oxidant effects. Thus, the balance between protection and toxicity depends on dose, exposure duration, and biological context [3,11].

4 Application Status of BHA

4.1 Applications in the Food Industry

The food industry is the primary sector for BHA use. It is commonly added to edible vegetable oils, animal fats, meat products, fried foods, and baked goods. According to China’s national standard GB 2760-2024, the maximum permitted addition level is 0.2 g/kg. At this concentration, BHA effectively suppresses increases in peroxide value and malondialdehyde_key indicators of rancidity. It can extend product shelf life by two to three times and significantly reduce storage losses [12]. In practice, BHA is usually applied in combination with other additives. Mixtures with BHT, propyl gallate, citric acid, or natural antioxidants (e.g., rosemary extract and tea polyphenols) leverage complementary mechanisms to achieve synergistic efficacy while mitigating safety risks [13,14]. Moreover, blends of BHA and ascorbyl palmitate have demonstrated notable synergistic effects in preventing oxidative degradation in food and pharmaceutical systems [15].

4.2 Applications in Polymer Materials

Owing to its excellent thermal and oxidative stability, BHA is often combined with phosphite auxiliary antioxidants and incorporated into polymers such as polyethylene, polypropylene, and rubber. During high-temperature processing and long-term service, BHA inhibits oxidative scission of polymer chains. This delays ageing, embrittlement, and mechanical property degradation, thereby extending the service life of plastic and rubber products by three to five years. It is widely used in anti-ageing modification of industrial plastics, rubber goods, packaging materials, and rubber accessories [16].

4.3 Applications in Cosmetics and Feed

In the cosmetics industry, BHA is used as an antioxidant preservative for oil-based products and skincare ingredients. In 2025, the EU Scientific Committee on Consumer Safety (SCCS) set a safe use concentration of 0.07% for leave-on and rinse-off cosmetics. BHA is prohibited in oral products and those with inhalation exposure risks. Normal skin contact does not cause notable irritation [4]. In animal feed, BHA at dosages up to 150 mg/kg effectively delays oxidation of feed oils. It accumulates minimally in livestock, supporting its wide acceptance [5].

5 In Vivo Metabolism and Environmental Behaviours of BHA

5.1 In Vivo Metabolic Pathways

Dietary intake is the main exposure route for the general population. BHA residues are rapidly absorbed through the digestive tract and predominantly metabolised in the liver. Its metabolic fate determines bioaccumulation and toxicity potential. In humans, metabolism proceeds mainly through conjugation: approximately 72% of BHA is converted to glucuronic acid conjugates, 14% to sulfate esters, and only a small fraction remains as the parent compound. Most metabolites are excreted in urine and faeces within 24 hours. Thus, at typical dietary levels, there is no significant accumulation risk. However, hepatic cytochrome P450 enzymes can demethylate BHA to generate TBHQ_a highly reactive intermediate. TBHQ is considered a key mediator of genetic toxicity and cellular damage [3].

From a kinetic perspective, single short-term exposure at approved doses is rapidly cleared. Nevertheless, long-term continuous low-dose exposure may lead to slow accumulation of toxic metabolites, disrupting metabolic homeostasis and causing chronic oxidative injury and multi-system pathologies. This is currently the major health concern associated with BHA [2].

5.2 Environmental Residues and Bioaccumulation

BHA and other synthetic phenolic antioxidants enter the environment via industrial wastewater, material migration, product residues from food processing, plastics manufacturing, and cosmetic use. Environmental monitoring has detected BHA in wastewater treatment plant influent and effluent, receiving rivers, sludge, sediments, and indoor dust, indicating their continuous release into the environment [2,17]. However, current risk assessments do not classify BHA as a highly persistent or strongly bioaccumulative pollutant. Its environmental fate is governed by photolysis, oxidation, and biodegradation. Some aquatic organism studies indicate that BHA can accumulate under specific exposure conditions, and co-exposure with microplastics may increase its internal burden and developmental toxicity. Nonetheless, robust evidence for significant biomagnification along food chains is lacking [18].

Recent theoretical and microbial studies suggest that BHA is not uniformly persistent across all environments. Quantum chemical and kinetic modelling predict that aqueous BHA can react with reactive oxidants_including hydroxyl radicals, sulfate radicals, and ozone_and that degradation rates vary widely with oxidation system and conditions [19]. In controlled anaerobic digestion, BHA was completely removed within 15 days, indicating strong biotransformation potential by anaerobic microbial communities. However, the specific degrading organisms and complete catabolic pathways remain to be elucidated [20].

6 Research Progress on Toxicological Safety of BHA

Current toxicological evidence indicates that BHA’s adverse effects exhibit three key characteristics: dose-dependency, species-specificity, and multi-target activity. Short-term exposure at regulatory levels is considered manageable. However, high-dose acute exposure, prolonged low-dose cumulative exposure, and combined exposure with other additives can induce multi-organ and multi-system injuries. Network toxicology analyses have revealed that BHA modulates core genes such as NFKB1, TNF, IL6, and ESR1, thereby perturbing normal physiological processes through multiple molecular pathways [21].

6.1 Controversies over Potential Carcinogenic Risk

Carcinogenic risk remains the central controversy. In 1986, IARC classified BHA as a Group 2B possible human carcinogen, based on rodent studies showing that high doses induced forestomach hyperplasia, papillomas, and squamous cell carcinomas in rats. The U.S. NTP also reported carcinogenic activity, sparking global debate [7]. Long-term feeding studies demonstrated that doses above 1200 mg/kg body weight consistently produce gastric tumours in rats and hamsters [6].

However, subsequent mechanistic research has clarified that this effect is highly species-specific. Felter et al. (2021) conducted a cross-species comparative study and found that the tumour-inducing mechanism in rats depends on their unique forestomach epithelium and specific enzyme expression. Human gastric physiology and metabolic pathways differ fundamentally, so this pathway is not activatable in humans [22]. To date, no epidemiological evidence has confirmed that dietary BHA at conventional levels causes cancer in humans. IARC itself acknowledges that the animal data cannot be directly extrapolated to humans. Therefore, under approved conventional use, no definite human carcinogenic risk exists.

6.2 Cellular and Genotoxic Effects

In vitro studies demonstrate concentration-dependent cytotoxicity and genotoxicity. At an environmentally relevant concentration of 50 μM, BHA impairs normal function of human astrocytes. At concentrations above 100 μM, it directly causes DNA fragmentation and chromosomal damage [23]. The liver, being the primary metabolic site, shows hepatocyte oedema, inflammatory infiltration, and abnormal function after long-term low-dose exposure. High doses induce pathological tissue damage. Animal models also confirm that high-dose BHA causes organic lesions in the stomach and kidneys of rats [24]. Network toxicology further indicates that BHA promotes apoptosis and tissue injury through synergistic multi-target and multi-pathway actions [21].

6.3 Neurotoxicity and Endoplasmic Reticulum Stress

Recent studies have extended the toxicity profile beyond the traditional focus on liver and kidney toxicity to the nervous system. Park et al. (2019) reported that low, environmentally relevant concentrations of BHA induce abnormal calcium accumulation in human astrocytes. This triggers sustained endoplasmic reticulum stress, interferes with neural cell proliferation, differentiation, and metabolic rhythms, and ultimately leads to neuronal apoptosis and dysfunction. These findings suggest potential risks of cognitive impairment and neurodevelopmental abnormalities from long-term low-dose exposure [23].

Zebrafish embryo assays further confirm developmental neurotoxicity. Exposure to BHA at concentrations as low as 0.5–8 ppb for 96 hours delays hatching, reduces heart rate, and causes spinal curvature, yolk sac oedema, and pericardial oedema. Concurrently, activities of antioxidant enzymes (CAT, GPx, SOD) decrease significantly. Combined oxidative and endoplasmic reticulum stress exacerbate neural damage. BHA also inhibits acetylcholinesterase activity, lowers serotonin levels, and downregulates key neurodevelopmental genes_including DRD4, COMT, and BDNF_leading to increased anxiety and impaired memory. This implies that fetuses and infants may be particularly vulnerable [25].

6.4 Bidirectional Nrf2 Regulation and Oxidative Stress

Oxidative stress imbalance is a core mechanism underlying BHA toxicity. Its bidirectional modulation of the Nrf2 pathway explains the dose-dependent effects. At low doses, BHA activates Nrf2 in mouse liver and small intestine, upregulating antioxidant and detoxifying enzymes and protecting against basal oxidative damage [11]. When the dose exceeds the safety threshold or exposure is prolonged, excessive reactive oxygen species accumulate, damaging cell membranes, mitochondria, and DNA. This disrupts redox homeostasis, triggers inflammation and cell death, and manifests as overt toxicity [11]. This “low-dose protection, high-dose toxicity” pattern provides a theoretical foundation for establishing safe application limits.

6.5 Endocrine Disruption and Obesogenic Effects

Isomer-specific effects on lipid metabolism have been observed. In 3T3-L1 cells, 3-BHA promotes adipocyte differentiation and lipid accumulation, whereas 2-BHA does not [1,9]. Additionally, 3-BHA interferes with the differentiation of mesenchymal stem cells into brown adipocytes [9]. In human HepaRG hepatocytes, BHA reduces DIO2 expression and affects thyroid hormone metabolism-related genes, such as UGT1A1, suggesting interference with the conversion of T4 to T3 and hepatic thyroid hormone handling [26]. Receptor and reporter gene assays have shown weak oestrogenic, anti-oestrogenic, and anti-androgenic activities [27,28]. High-dose animal studies observed thyroid alterations, changes in sex hormone levels, and impaired reproductive development. However, it remains unclear whether such effects occur at typical dietary exposure levels [29].

6.6 Combined Exposure and Epidemiological Evidence

Single-compound assessments have inherent limitations. Recent population-based studies have focused on mixture effects. In 2025, Wang et al. investigated a normoglycemic rural Chinese population and found that combined exposure to BHA and BHT significantly interfered with sex hormone levels [30]. These effects were gender-specific and modified by body mass index. In males, mixed exposure positively correlated with progesterone and negatively correlated with testosterone and androstenedione. The endocrine-disrupting effects were amplified in overweight or obese individuals. This provides the first population-level evidence that BHA may pose health risks in real-world, multi-exposure scenarios.

7 Comparison of Global Regulatory Standards for BHA

Given the well-documented multi-system toxicity, regulatory authorities worldwide have progressively tightened controls. The general trends include lower limits, narrower scopes of use, and stricter restrictions for vulnerable groups.

·China: GB 2760-2024 sets a maximum addition level of 0.2 g/kg in all food categories. BHA is prohibited in infant foods and medical formula foods. Residue limits in nutritional supplements are strictly set at ≤0.2 mg/kg. Domestic exposure monitoring indicates that daily intakes are generally below safety thresholds, so population health risks are considered low [31].

·European Union: BHA is classified as a regulated endocrine-disrupting substance. EFSA has established an Acceptable Daily Intake (ADI) of 1 mg/kg bw. BHA is completely banned in infant complementary foods and special dietary foods. Its use in food and feed is tightly restricted. SCCS has set a safe concentration of 0.07% in cosmetics and prohibits its use in oral and inhalable products [4,6].

·United States: The FDA still lists BHA as GRAS, with a food addition limit of 0.02%. It allows limited use in conventional foods but continuously monitors carcinogenic and endocrine risks. The comprehensive safety review launched in 2026 may reshape the U.S._and potentially the global_regulatory framework [7,8].

·Japan: BHA is permitted in specific foods_butter, oils, dried or salted fish and shellfish, and dehydrated potato puree_with a general limit of 0.2 g/kg. For certain frozen fish, shellfish, and whale meat, the limit in impregnating liquid is 1 g/kg. When BHA is used together with BHT, the total amount must not exceed the respective limits [32].

8 Cutting–Edge Research Directions

8.1 Toxicity Reduction via Natural–Synthetic Combinations

Combining BHA with natural antioxidants is currently the most effective strategy to balance efficacy and safety. Mixtures with rosemary extract, pomegranate peel extract, liposoluble tea polyphenols, or citric acid achieve synergistic effects. This can reduce the BHA dosage by more than 50% while maintaining antioxidant performance, thereby lowering toxic exposure [13,14]. Studies indicate that 1000 ppm pomegranate peel extract outperforms traditional BHA/BHT blends. Natural antioxidants derived from agricultural by-products show strong substitution potential [33]. Additionally, BHA with ascorbyl palmitate exhibits synergistic inhibition of oxidative degradation in food and pharmaceuticals [15]. This formulation approach is now the mainstream optimisation direction for the food industry’s use of BHA.

8.2 Molecular Structure Modification for Lower Toxicity

Using molecular docking and quantum chemical calculations, researchers are modifying the substituent groups on BHA’s benzene ring to retain the antioxidant core while weakening toxic sites. The resulting derivatives have comparable antioxidant efficiency but significantly lower cytotoxicity and genotoxicity, avoiding the safety concerns of parent BHA. This represents a promising avenue for developing novel low-toxicity antioxidants.

8.3 Nano–Carrier Slow–Release and Toxicity Control

Encapsulating BHA in biocompatible carriers_such as cyclodextrin or chitosan_enables targeted, slow, and controlled release [16]. This improves thermal and storage stability, prevents local high-concentration toxicity from rapid release, reduces direct absorption and accumulation, and substantially lowers cytotoxicity and chronic injury risks. This technology holds strong potential for high-end food, pharmaceutical, and cosmetic applications.

8.4 Refined Exposure Risk Assessment

Risk assessment has moved from acute high-dose testing to differentiated evaluation covering long-term low-dose exposure, multi-additive mixtures, and specific subpopulations. Network toxicology, gut microbiome omics, and population follow-up data are being integrated to construct multi-target, multi-pathway toxicological profiles and improve refined risk databases [21]. Furthermore, isomer-specific risk assessment distinguishes the safety thresholds of 3-BHA and 2-BHA, enabling more precise regulation–a core future trend [9].

9 Conclusions and Prospects

9.1 Research Conclusions

BHA offers irreplaceable advantages in high-temperature processed foods, polymers, cosmetics, and feed, owing to its potent antioxidant activity and thermal stability. It effectively delays oxidative deterioration and reduces industrial losses. However, its toxic effects are dose-dependent, species-specific, and multi-target. Short-term use at approved doses is considered safe, with no confirmed human carcinogenic risk. Nevertheless, long-term cumulative low-dose exposure and combined exposure with other additives can cause DNA damage, neurodevelopmental toxicity, and endocrine disruption via oxidative stress, endoplasmic reticulum stress, and calcium imbalance. BHA is metabolised in the liver to the highly toxic intermediate TBHQ, which is a key driver of biological damage. BHA also occurs in the environment and may bioaccumulate. Although it is degradable through advanced oxidation and microbial pathways, the ecological and health risks cannot be overlooked. Globally, regulatory standards are being tightened. Strategies such as combined formulations, nano-encapsulation, and structural modification effectively balance application value and safety, and represent the current best approaches for optimising BHA use.

9.2 Development Prospects

Future research and industrial application will focus on three core directions. First, promote low-toxicity molecular modification to generate highly active, easily metabolised, low-residue BHA derivatives, while avoiding “regrettable substitution” and conducting full life-cycle safety assessments for new alternatives. Second, establish standardised combination systems, develop quantitative models linking dosage, efficacy, and toxicity, and formulate industrialisable recipes that enhance efficacy and reduce toxicity. Third, deepen population toxicology research on long-term low-dose combined exposure, integrate multi-omics technologies to refine risk assessment, and facilitate global regulatory harmonisation. As green consumption and environmental policies gain momentum, BHA will evolve towards more refined, low-toxicity, and precise use. Substitution with natural antioxidants such as liposoluble tea polyphenols will become a long-term industry trend. The FDA’s 2026 comprehensive safety reassessment may reshape the global regulatory and industrial landscape for BHA.

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