IngredientsBenzyl isothiocyanate (commonly abbreviated BITC) is a member of the isothiocyanate (ITC) class of organosulfur compounds. It is an isothiocyanate found in plants of the mustard family. Its systematic IUPAC name is (isothiocyanatomethyl)benzene; it is also known in the literature and trade as benzyl mustard oil, Ī±-isothiocyanatotoluene, benzylsenfoel (German), and by the CAS registry number 622-78-6. The molecular formula is CāHāNS, with a molecular weight of approximately 149.2 g/mol.
It is a colorless to pale yellow liquid with a boiling point of 242ā243 Ā°C and a characteristic watercress-like odor. At the 1.0% level, the odor has been described as vegetative, cabbage and radish-like, green with a musty sulfurous earthiness reminiscent of mustard greens; at 0.5 ppm the taste is green, vegetative, cabbage-like, kimchi, radish, and wasabi-like, imparting a strong trigeminal tongue-numbing and raspy sensation.
BITC does not exist freely in intact plant tissue. Glucotropaeolin (benzyl glucosinolate) is a glucosinolate found in cruciferous vegetables, particularly garden cress; upon enzymatic activity, it is transformed into benzyl isothiocyanate, which contributes to the characteristic flavor of these brassicas. Glucotropaeolin is biosynthesised from the amino acid phenylalanine in a multi-step pathway. The compound was first reported in 1899, after its isolation from Tropaeolum majus, a nasturtium species. Glucotropaeolin is now known to occur widely in other brassica families including Caricaceae, Phytolaccaceae, Resedaceae, Salvadoraceae, and Tovariaceae.
A range of organic isothiocyanates, including benzyl isothiocyanate, occur as secondary metabolites in plants, most notably in species of the Brassicaceae family. Key sources identified in the peer-reviewed literature include:
These ITCs form part of our daily diet and are synthesized in the plant from chemically rather unreactive glucosinolates (Ī²-thioglucoside-N-hydroxysulfates) in an enzymatic conversion involving myrosinase enzymes. When plant tissues are damaged, the endogenous enzyme myrosinase (Thioglucoside hydrolase, EC 3.2.3.1), stored in myrosinase grains of the myrosin cells, is released and combined with glucosinolates to produce biologically active compounds, such as nitriles, thiocyanates, isothiocyanates, epithionitriles, and oxazolidine-2-thiones. During the cooking process of cruciferous vegetables, myrosinase activity and associated protein specifier proteins are usually destroyed unless strict cooking times are adhered to; despite the thermal destruction of plant myrosinase activity, the intake of cooked Brassica vegetables still results in the formation of bioactive isothiocyanates and nitriles, which arise from the metabolism of glucosinolates by the human gut microbiota.
According to the European Prospective Investigation into Cancer and Nutrition (EPIC), the average consumption of aromatic glucosinolates such as benzyl glucosinolate varies between European countries: 1.4 mg/day (men) to 0.74 mg/day (women) in the German population, and 4.46 mg/day (men) to 4.06 mg/day (women) in the Spanish population, suggesting differently high but regular intake of Brassicales species.
Throughout the world, 182 species of plants have been used as sources for chewing sticks, of which the roots of the shrub Salvadora persica is the most common one; its use remains widespread in many parts of the world from East Africa through to the Asian subcontinent. Sticks from the roots of S. persica, Miswak sticks, have been used for centuries as a traditional method of cleaning teeth. Miswak or Siwak (Salvadora persica) is known long back in history for use as a natural toothbrush for oral hygiene; based on the tradition inherited from Prophetic medicine, the use of Siwak is widespread in Islamic countries. In addition to the mechanical removal of dental plaque and stimulation of the gingival tissues, early reports indicated that the root of S. persica exerts antimicrobial activity. It is now established that BITC is the principal antimicrobially active component responsible for these properties.
Tropaeolum majus L., an herbaceous plant commonly known as garden nasturtium, belongs to the family Tropaeolaceae and is native to Peru; it was first introduced to Europe in the sixteenth century and then spread to other parts of the world, including malaria-endemic countries such as Angola, Rwanda, and Vietnam. T. majus was selected as a potential candidate for antimicrobial drug development not only because of its widespread distribution but also because of its traditional usages against bacterial infections such as bronchitis, sinusitis, and urinary tract infections, as well as for its antifungal and antiviral activities. A combination of BITC, allyl isothiocyanate (AITC), and phenylethyl isothiocyanate (PEITC) forms the active principles of ANGOCINĀ® Anti-Infekt N (Angocin; film-coated tablets, Repha GmbH, Langenhagen, Germany), a therapy for managing inflammatory diseases of the respiratory and urinary tracts.
Papaya (Carica papaya) seeds have a long history of use in many tropical regions as a traditional anthelmintic and digestive remedy. BITC was identified scientifically as the chief or sole anthelmintic component in papaya seed extracts (Kermanshai et al., 2001, Phytochemistry 57:427ā435). Benzyl isothiocyanate, the main breakdown product of benzyl glucosinolate, is present in all parts of Tropaeolum majus L. and has antibacterial and antiparasitic activities.
Once formed, ITCs react readily with thiol and amine groups present in biomolecules; they are able to modify and hence regulate key proteins and enzymes, which results in widespread cellular responses, including the expression of Phase II enzymes. This electrophilic characterāthe isothiocyanate functional group (āN=C=S)āis central to nearly all of BITC's known biological activities.
Isothiocyanates are potent inducers of Phase II enzymes in vitro, and they have been shown to increase the metabolism and detoxification of chemical carcinogens in vitro and in animal models. Glucosinolate hydrolysis products, especially isothiocyanates, are potent inducers of Phase II detoxifying enzymes and subsequently confer protection against oxidative stress and chronic inflammation. Prooxidants or electrophiles, including BITC, may directly interact with cysteine residues present in Keap1, thereby stimulating Nrf2 dissociation; both events can facilitate the nuclear translocation of Nrf2, which subsequently associates with small Maf, forming a heterodimer that binds to the antioxidant-response element (ARE) or electrophile-responsive element (EpRE) to stimulate Phase II detoxification or antioxidant enzymes.
Specifically for BITC, BITC induces Phase II enzymes, increasing levels of heme oxygenase 1 (HO-1), glutathione, and glutamate cysteine ligase; it also decreases levels of ROS, activation of NF-ĪŗB, adhesion of monocytes, and expression of ICAM-1, VCAM-1, and E-selectin.
BITC-induced apoptosis in human breast cancer cells is initiated by reactive oxygen species (ROS) due to inhibition of complex III of the mitochondrial respiratory chain. Previous studies have shown that BITC produces a large number of intracellular reactive oxygen species (ROS) and increases intracellular CaĀ²āŗ release from endoplasmic reticulum (ER), leading to the activation of the apoptotic mechanism in tumor cells.
BITC exerts anticancer efficacy through modulation of various signaling pathways involved in apoptosis, cell proliferation, cell cycle arrest, metastasis, angiogenesis, and autophagy. BITC-mediated G2/M arrest was associated with up-regulation of cyclin dependent kinase inhibitor p21(Waf1/Cip1) and the activation of checkpoint kinase 2, whereas the expressions of other G2/M regulatory proteins, including CyclinB1, Cdc2, and cell division cycle 25C (Cdc25C), were down-regulated by 19, 51, and 70%, respectively, compared with control.
BITC causes cell cycle arrest at G2/M phase, reactive oxygen species generation, and glutathione depletion; Akt activity and NF-ĪŗB transcriptional activation were suppressed, while mitogen-activated protein kinase and activator protein 1 (AP-1) were activated.
The antibacterial property of benzyl isothiocyanate is owing to its chemical structure, as it has both lipophilic and electrophilic properties which render it able to penetrate through the outer bacterial membrane and disturb the ability of bacteria to maintain cellular integrity. The transcriptomic response of C. jejuni to benzyl isothiocyanate showed upregulation of heat shock response genes and an impact on energy metabolism; this study demonstrates that exposure to BITC elicits a heat shock response and an oxidative stress response and induced the expression of genes involved in energy production and electron transport.
BITC is one of the common isothiocyanates found in cruciferous vegetables such as broccoli, cabbage, or watercress; preclinical studies report its effectiveness in the prevention and treatment against several cancers; one review aimed to report and discuss findings on anticancer activities of BITC and its modes of action against 14 types of cancer. A total of 75 publications regarding the anticancer properties of BITC were included in one systematic review, and the authors concluded that further studies are needed to validate its effectiveness in humans for further development and translation into prophylaxis or therapy. Evidence to date is predominantly preclinical (in vitro and animal models); no completed large-scale human clinical trials specifically assessing BITC monotherapy for cancer endpoints have been identified in the available literature.
BITC has been shown to inhibit chemically induced pancreatic cancer in experimental animals; in one study, BITC (10 Āµmol/L) treatment caused marked phosphorylation of H2A.x (2.6-fold) and permanent damage to Capan-2 human pancreatic cancer cells. In 2006, there was a report that isothiocyanates, including benzyl isothiocyanate, protected against pancreatic carcinogenesis in vitro. These findings are preclinical only.
BITC, described as a dietary cancer chemopreventive agent, causes apoptosis in MDA-MB-231 and MCF-7 human breast cancer cells, though the mechanism of cell death was not fully understood at the time of that study. BITC treatment inhibits complex III of the mitochondrial respiratory chain in breast cancer cells to trigger generation of ROS, which function upstream of JNK and p38 MAPK activation and mitochondrial translocation (activation) of Bax in BITC-induced apoptosis. All studies are in vitro or animal models.
BITC inhibited gefitinib-resistant human non-small-cell lung cancer (NSCLC) cell growth by inducing apoptosis in a dose-dependent manner and activated caspase-3, with no effects on epidermal growth factor receptor and multidrug resistant protein expression. These results demonstrated that BITC overcame gefitinib resistance in lung cancer cells. This is an in vitro finding only.
BITC is a cruciferous vegetable-derived compound with anticancer properties in human cancer cells; in oral cancer cells (OC2), BITC inhibited growth, promoted G2/M phase arrest, and triggered apoptosis with minimal toxicity to normal cells. BITC rapidly produced reactive oxygen species and nitric oxide, triggering oxidative DNA damage; BITC effectively decreased the intracellular GSH and GSH/GSSG ratio. A separate study in cisplatin-resistant oral cancer cells (CAR cells) found that BITC represents a promising candidate as an adjuvant treatment for oral anticancer, and it might be a potential agent for patients with drug-resistant oral cancer in the future. These findings remain preclinical.
Anaplastic thyroid carcinoma (ATC) is the most aggressive type of thyroid cancer; BITC is a natural compound that has shown promising anticancer properties, and one study aimed to evaluate the antitumor effect of BITC in ATC, highlighting signaling pathways involved, via in vitro and in vivo studies. Again, this represents preclinical evidence only.
BITC induces G2/M phase arrest and apoptosis in human melanoma A375.S2 cells through reactive oxygen species (ROS) and both mitochondria-dependent and death receptor-mediated multiple signaling pathways. This finding is in vitro.
BITC is a member of ITCs with a benzene ring side face and short hydrogen chain; several studies have provided evidence that the bactericidal effect of BITC is stronger than other ITCs against pathogens such as Methicillin-Resistant Staphylococcus aureus (MRSA) strains, Candida albicans, and Aspergillus niger. In an in vitro study examining MRSA isolates, BITC at 440 Āµg/disc produced inhibitory zones (10.3 Ā± 0.6 mm against the reference strain) against all MRSA isolates tested, though vancomycin at 30 Āµg/disc produced larger zones (15.7 Ā± 0.6 mm), indicating BITC's antibacterial activity against drug-resistant organisms is real but does not exceed that of established antibiotics at the tested concentrations.
BITC exhibited potent antimicrobial activity under in vitro conditions; in a comparative mouse model study of BITC with gentamycin sulfate and ceftiofur hydrochloride against Pseudomonas aeruginosa infection, results showed that BITC exhibited comparable or better antimicrobial activity and lower infiltration of mouse immune cells compared to gentamycin sulfate; furthermore, BITC did not impose any toxicity to the air pouch skin tissues. Most antibacterial studies on BITC were undertaken under in vitro conditions. This in vivo mouse study is one of few preclinical animal studies assessing BITC antibacterial activity in a live infection model.
BITC exerted antifungal effects against Aspergillus fumigatus by damaging cell membranes, mitochondria, adhesion, and biofilms in a concentration-dependent manner; in vivo (mouse model), fungal load and inflammatory response including inflammatory cell infiltration and pro-inflammatory cytokine expression were reduced in BITC-treated A. fumigatus keratitis. This represents preclinical animal evidence.
Oxygen consumption was progressively impaired by benzyl isothiocyanate treatment, as revealed by high-resolution respirometry, while the ATP content increased soon after benzyl isothiocyanate exposition, suggesting a shift in the energy metabolism balance in C. jejuni. This is in vitro/mechanistic evidence.
The most clinically developed application of BITC in humans is as part of the standardized combination herbal product ANGOCINĀ® Anti-Infekt N (Repha GmbH, Germany), in which nasturtium (T. majus) and horseradish (Armoracia rusticana) are combined. Angocin comprises horseradish root (80 mg) and nasturtium (200 mg), with respective ITC ratios of PEITC (12%), AITC (38%), and BITC (50%); AITC and PEITC originate from horseradish root, while nasturtium is a rich source of BITC.
Combined in the medicinal product ANGOCINĀ® Anti-Infekt N, the two natural components demonstrated promising effects against acute bronchitis; a randomized, two-armed, placebo-controlled, double-blind, Phase IV study revealed the healing-fostering effect of the two herbal plant components in a study that included 384 patients, with 195 in the treatment and 189 in the placebo group, using the "bronchitis severity score" (BSS) as primary endpoint.
A randomized, double-blind, placebo-controlled trial demonstrated the efficacy and safety of the herbal medicinal product Angocin Anti-Infekt N in the prophylactic treatment of chronically recurrent UTIs. In nasturtium, benzyl isothiocyanate is the corresponding mustard oil. The effectiveness of this substance in reducing inflammation has been established in vitro as well as in numerous clinical trials, primarily owing to the presence of ITCs.
Limitation: Because BITC is combined with AITC and PEITC in ANGOCINĀ®, human clinical evidence cannot be attributed to BITC alone. No human clinical trial isolating BITC as a single agent for infection has been identified in the available peer-reviewed literature.
Only few studies have linked isothiocyanates with type 2 diabetes (T2D) prevention; sulforaphane supplementation from broccoli sprouts was shown to reduce insulin, inflammatory markers, and LDL levels in T2D patients, although the mechanisms underlying these effects are not clear. Also Moringa extracts rich in 4(Ī±-L-rhamnosyloxy)-benzyl isothiocyanate (a BITC analogue) were shown to display anti-obesity and anti-diabetic properties increasing insulin signaling and sensitivity. One published study showed BITC's restraining impact on obesity, fatty-liver, and insulin resistance in diet-induced obesity mouse models. Evidence for BITC specifically in this area is currently limited to in vitro cell studies and animal models; no completed human trials for these outcomes were identified.
BITC, the main breakdown product of benzyl glucosinolate, is present in all parts of Tropaeolum majus L. and has antibacterial and antiparasitic activities; to date, there is no information on the effects of BITC specifically against malaria. One 2024 study in Molecules evaluated the antimalarial activity of BITC against Plasmodium falciparum cultures in vitro. Researchers used flow cytometry to calculate growth inhibition percentage against unsynchronized cultures of the chloroquine-susceptible P. falciparum 3D7-GFP strain, validating extracts with at least 70% growth inhibition by IC50 estimation against both 3D7-GFP and Dd2 (chloroquine-resistant strain) cultures. This is an early-stage in vitro study.
The long-established traditional use of miswak (Salvadora persica) as a dental hygiene aid now has a scientific basis in BITC as the primary active compound. In addition to the mechanical removal of dental plaque and stimulation of the gingival tissues, there were early reports indicating that the root of S. persica exerts antimicrobial activity. The identification of BITC as the main active antimicrobial component was confirmed by gas chromatographyāmass spectrometry studies (Sofrata et al., 2011, PLOS ONE). Clinical data comparing miswak to standard toothbrushing exist in the oral hygiene literature, but these are attributed to the whole plant preparation rather than isolated BITC.
No official dose monograph exists specifically for isolated BITC as a dietary supplement. The doses below are drawn directly from specific studies as reported.
The corresponding cysteine conjugate was formed when the glutathione (GSH) or cysteinylglycine conjugates of benzyl isothiocyanate were incubated with rat liver or kidney homogenates; when the cysteine conjugate of benzyl isothiocyanate was similarly incubated in the presence of acetyl-CoA, the corresponding N-acetylcysteine conjugate (mercapturic acid) was formed. The mercapturic acid was excreted in the urine of rats dosed with benzyl isothiocyanate or its GSH, cysteinyl-glycine, or cysteine conjugates. An oral dose of the cysteine conjugate of [Ā¹ā“C]benzyl isothiocyanate was rapidly absorbed and excreted by rats and dogs; after 3 days, rats had excreted a mean of 92.4% and 5.6% of the dose in the urine and faeces respectively, and dogs had excreted a mean of 86.3% and 13.2% respectively.
In humans, the majority of biologically active glucosinolate breakdown products are eliminated through urine as mercapturic acid derivatives, with a smaller fraction exhaled as volatile ITCs and nitriles. ITCs could bind to proteins either partially or irreversibly, potentially reducing the bioavailability of glucosinolates when consumed with protein-rich foods. The chemical structure of benzyl glucosinolate present in nasturtium and the corresponding BITC have been characterized analytically, and the metabolism via the mercapturic acid pathway in human beings has been shown in detail.
BITC is very irritant to tissues in its isolated, concentrated form. At the doses encountered from dietary cruciferous vegetable consumption, this is of minimal practical concern, but it is highly relevant for concentrated preparations or occupational exposure to the pure compound.
An important preclinical safety signal concerns the urinary tract. Allyl isothiocyanate (AITC) is known to be weakly carcinogenic, whereas benzyl isothiocyanate (BITC) has been suggested to exert carcinogenicity toward the rat urinary bladder. In rat intravesical instillation studies, BITC caused more profound toxic damage than AITC; among the BITC-metabolites, cytotoxicity was evident with intermediate glutathione or cysteinylglycine conjugates, whereas the mercapturic acid, considered to be the major final urinary metabolite, exerted little effects. This toxicity was demonstrated via direct intravesical instillation in rodent models at 2.8 mg/kg body weight, a route and dose not representative of normal dietary or oral supplemental exposure; nevertheless, the finding has led researchers to note caution at supra-physiological concentrations.
In studies using a normal renal proximal tubular cell line (pig LLC-PK1), BITC increased cell death with an ICā ā value of about 7 ĀµM, whereas the cytotoxic effect of BITC-NAC (its mercapturic acid metabolite) was five times weaker; significant necrosis with oxidative stress was observed; and in the presence of 5 mM glutathione (comparable to physiological levels), the cytotoxicity of both BITC-NAC and BITC was significantly reduced. This finding suggests glutathione status may modulate BITC cytotoxicity in vivo.
ITCs could bind to proteins either partially or irreversibly, potentially reducing the bioavailability of glucosinolates when consumed with protein-rich foods. This is a pharmacokinetic interaction relevant to dietary intake scenarios.
Despite the thermal destruction of plant myrosinase activity during cooking, intake of cooked Brassica vegetables still results in the formation of bioactive isothiocyanates and nitriles, which arise from the metabolism of glucosinolates by the human gut microbiota. This implies that microbial community composition may affect the amount of BITC bioavailable from dietary sources, and that BITC in turn may influence the gut microbiome, though specific clinical data on this interaction require further research.
BITC, which possesses antitumor activity as an alkylating agent, reacts with D-glucosamine (commonly employed in oral treatments against osteoarthritis and inflammation); chemical results suggest that clinical treatments with D-glucosamine could reduce the beneficial effects associated with diets based on glucosinolate-rich foods. This is a biochemically characterized potential interaction, but has not been confirmed in human pharmacokinetic studies.
The two most recent clinical trials investigating the efficacy and safety of ANGOCINĀ® therapy in acute rhinosinusitis and acute bronchitis proved that the product is safe and well-tolerated. These findings are specific to the licensed combination product, not to isolated BITC.
Health conditions that Benzyl isothiocyanate may help support.
Body systems that Benzyl isothiocyanate may help support.
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