Absorption, Distribution and Excretion
…When rats were fed vanillin at a dose of 100 mg/kg, most metabolites were excreted in the urine within 24 hours…

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Metabolism/Metabolites …After intraperitoneal injection of vanillin into rats, various metabolites were produced in the urine; the most important of these were free and bound vanillic acid. Other metabolites included bound vanillin, bound vanillyl alcohol, and catechols. Protocatechuic aldehydes may be produced by microorganisms; vanillin-4-β-D-glucoside is produced in legumes and japonica rice; vanillin-4-β-D-glucuronide is produced in rabbits.
/Table Data/ …When rats were fed vanillin at a dose of 100 mg/kg, most metabolites were excreted in the urine within 24 hours, primarily as glucuronide and/or sulfate conjugates, but the generated acids were also excreted in free form and as glycine conjugates. Within 48 hours, 94% of the dose was excreted, including 7% vanillin, 19% vanillyl alcohol, 47% vanillic acid, 10% vanillyl glycine, 8% catechol, 2% 4-methylcatechol, 0.5% guaiacol, and 0.6% 4-methylguaiacol. Vanillic acid is produced in the human body. /Table Data/

Toxicity Summary
Identification: Vanillin is used in small amounts in perfumery to soften and fix sweet and balsamic aromas. It is also used as a whitening agent in electroplating processes and is an important intermediate in the production of drugs such as L-DOPA and methyldopa. Vanillin (EPA/OPC Pesticide Code: 115801) currently has no matching label. / Not registered for use in the U.S., but approved pesticide uses may change periodically, so consult federal, state, and local authorities for currently approved uses. / Pharmaceutical adjuvant (flavoring agent). Used as a flavoring agent in confectionery, beverages, food, and animal feed. Fragrances and flavoring agents in cosmetics. Synthetic reagent. Human Exposure and Toxicity: In a closed patch study on human skin, vanillin was tested at concentrations of 20%, 2%, and 0.4% in 29 healthy subjects, 30 healthy subjects, and 35 patients with dermatological conditions, respectively, without causing primary irritation. A maximum dose study was conducted in an experimental group of 25 volunteers. The substance was tested in petrolatum solution at concentrations of 2% and 5%, and no sensitizing reaction was observed. Vanillin is considered a minor allergen because sensitizing reactions have only been found in patients allergic to vanilla, isoeugenol, and coniferyl benzoate. Vanillin/ethyl vanillin may interact with drugs metabolized by CYP2E1/CYP1A2. Animal studies: Intraperitoneal injection of vanillin into strain A mice at total doses of 3.6–18.0 g/kg for 24 weeks did not result in excessive lung tumor development and is therefore not considered carcinogenic. Sixteen rats were divided into several groups and fed a diet containing vanillin at a dose of 20 mg/kg body weight/day for 18 weeks without any adverse reactions; however, a dose of 64 mg/kg/day for 10 weeks resulted in growth inhibition and damage to the myocardium, liver, kidneys, lungs, spleen, and stomach. In yeast experiments, vanillin acted as a co-mutagenic agent. Using cultured Chinese hamster V79 cells, the effects of vanillin on cytotoxicity and 6-thioguanine (6TG) resistance mutations induced by two different chemical mutagens—ethyl methanesulfonate (EMS) and hydrogen peroxide (H₂O₂)—were investigated. This study also examined the effect of vanillin on H₂O₂-induced chromosomal aberrations. When cells were co-treated with vanillin, vanillin showed a dose-dependent enhancement of EMS-induced cytotoxicity and 6TG resistance mutations. Posttreatment with vanillin also showed an increased frequency of EMS-induced mutations during the period of mutational expression following EMS treatment. However, when cells were post-treated with vanillin after H₂O₂ treatment, vanillin inhibited H₂O₂-induced cytotoxicity. Vanillin showed no effect in the absence of mutagenic activity in H₂O₂. Posttreatment with vanillin also inhibited H₂O₂-induced chromosomal aberrations. The differential effects of vanillin may be related to the nature of mutagenic DNA damage, and vanillin may affect at least two different cellular repair functions. Studies have found that vanillin (200 μg/culture) directly inhibits the in vitro anti-sheep erythrocyte antibody response at non-cytotoxic doses.

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Toxicity Data
LC (Rat)> 41.7 mg/m3/4hr

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Interactions
Methotrexate (MTX) is a chemotherapeutic drug used to treat cancer that causes cellular genetic damage and exhibits cytotoxic effects in various assay systems. Several antigenotoxic agents have been studied in various in vitro and in vivo systems. However, data on their ability to modulate MTX-induced genotoxicity are limited. In this study, we used vanillin (VA) and chlorophyll (CHL) as antigenotoxic agents to investigate their ability to reduce MTX-induced DNA damage. V79 Chinese hamster lung cells in the exponential growth phase were treated with five different concentrations (5-100 μg/mL) of MTX for 6 hours to activate S9 cells, followed by treatment with two concentrations of VA (50 or 100 μg/mL) or CHL (50 or 100 μg/mL) for 40 hours. Micronucleus (MN) assays were performed with cytochalasin B, and antigenotoxic agents were added to assess micronuclei in binucleated cells. Chromosomal aberrations were assessed in parallel cultures. Results showed that MTX alone dose-dependently reduced the mitotic index (NDI) and mitotic index (MI). A significant increase in the percentage of micronucleated binucleated cells (MNBN) and the percentage of abnormal cells (Abs) was observed. Studies using vanillin (VA) as an antigenotoxic agent showed that the addition of 50 or 100 μg/mL VA decreased both the number of MNBNs (26.3-83.1%) and the number of antibodies (16.0-87.5%). The addition of CHL also significantly reduced the number of MNBNs (53.0-91.5%), and this phenomenon was observed at both concentrations tested. Chromosomal aberration rates were also significantly reduced (41.0-83.0%). These studies indicate that both VA and CHL can effectively reduce MTX-induced chromosomal damage. Vanillin (VA) is an anti-chromosomal breakage agent that has been shown to inhibit gene mutations in bacterial and mammalian cells. However, data on its role in combating radiation-induced cellular genetic damage are limited. This study aimed to investigate the protective effect of VA against radiation-induced chromosomal damage in V79 cells. Cells in the exponential growth phase were exposed to five different doses of X-rays (1-12 Gy) and ultraviolet light (50-800 x 10² μJ/cm²), followed by treatment with three concentrations of VA (5, 50, or 100 μg/mL) for 16 hours (for micronucleus (MN) analysis) or 18 hours (for structural chromosomal aberration (SCA) analysis). MN and SCA assays were performed simultaneously according to standard procedures. Results showed that X-ray treatment alone increased the percentage of micronucleated binucleated cells (MNBNs) (5.6% to 79.6%) and abnormal cells (Abs) (12% to 98%) with increasing dose. Inhibition experiments showed that the addition of 100 μg/mL VA significantly reduced the percentage of micronucleated binucleated cells induced by X-ray irradiation 1, 2, and 4 times (21% to 48%). At VA concentrations of 5 and 50 μg/mL, the percentage of micronucleated binucleated cells decreased slightly. All three concentrations of VA reduced the percentage of antibodies induced by X-rays at all doses (15.7% to 57.1%). UV radiation alone significantly increased the percentage of micronucleated binucleated cells (3.5% to 14.8%) and antibodies (17% to 29%). The addition of 50 or 100 μg/mL VA significantly reduced the percentage of micronucleated binucleated cells (31.7% to 86.2%) and antibody percentage (54.5% to 90.9%) at all UV doses. Decreases in the percentage of micronucleated binucleated cells (2.8% to 72.4%) and antibody percentage (34.8% to 66.7%) were also observed at a VA concentration of 5 μg/mL. These data clearly demonstrate that VA has a protective effect against radiation-induced chromosomal damage, suggesting that VA is an anti-fracture agent. This study investigated the effects of dietary bioantimutagenic agents (compounds that have been shown to inhibit mutations by interacting with DNA repair processes) on the frequency of spontaneous and heterocyclic amine (HCA)-induced micronuclei (MN) in metabolically healthy human hepatocellular carcinoma cells (Hep-G2). All tested compounds (coumarin, vanillin, caffeine, tannic acid, and cinnamaldehyde) moderately increased the number of micronuclei in Hep-G2 cells at high concentrations (500 μg/ml); only tannic acid was active at lower doses. In experiments using the heterocyclic amine 2-amino-3-methylimidazo[3,4-f]quinoline (IQ), treatment of cells with the antimutagenic agent significantly reduced the number of micronuclei (75-90%). Vanillin, coumarin, and caffeine showed the most significant effects at concentrations ≤5 μg/ml. Further experiments showed that these compounds also attenuated the mutagenic effects of other heterocyclic amines (PhIP, MeIQ, MeIQx, Trp-P-1).
Injection of N-methylnitrosourea (MNU) into pregnant mice during early embryonic development (blastocyst formation) may induce congenital abnormalities by acting directly on the embryo. This study investigated whether several antimutagenic agents, including vanillin (VA) and cobalt chloride (CoCl2), could alter the developmental toxicity of N-methylnitrosourea administered to pregnant mice during the preimplantation period. On day 2.5 of gestation, ICR mice were intraperitoneally injected with a single dose of 20 mg/kg N-methylnitrosourea. One hour after N-methylnitrosourea treatment, mice were either intraperitoneally injected with a single dose of 50 mg/kg vanillin or intravenously injected with a single dose of 10 mg/kg cobalt chloride. Embryotoxicity and teratogenicity were assessed on day 18 of gestation. Cobalt chloride (CoCl2) significantly reduced N-methylnitrosourea-induced embryo/fetal mortality and markedly reduced the incidence of N-methylnitrosourea-induced external malformations. These inhibitory effects of vanillin and cobalt chloride support the view that the embryotoxicity of N-methylnitrosourea is caused by its direct effects on the embryo. Furthermore, the effects of vanillin and cobalt chloride are considered to be direct modifications of the affected embryos following N-methylnitrosourea treatment. ...
This study used cultured Chinese hamster V79 cells to investigate the effects of vanillin on cytotoxicity and 6-thioguanine (6TG) resistance mutations induced by two different chemical mutagens—ethyl methanesulfonate (EMS) and hydrogen peroxide (H2O2). The effect of vanillin on hydrogen peroxide-induced chromosomal aberrations was also examined. The results showed that vanillin had a dose-dependent enhancing effect on EMS-induced chromosomal aberrations. Both cytotoxicity and 6TG resistance mutations were increased when cells were simultaneously treated with vanillin. Posttreatment with vanillin during the mutation expression period after EMS treatment also showed an increased EMS-induced mutation frequency. However, when cells were posttreated with vanillin after hydrogen peroxide treatment, vanillin inhibited hydrogen peroxide-induced cytotoxicity. Vanillin showed no effect when hydrogen peroxide did not have mutagenic activity. Posttreatment with vanillin also inhibited hydrogen peroxide-induced chromosomal aberrations. The differential effects of vanillin may be related to the nature of mutagenic DNA damage, and vanillin may affect at least two distinct cellular repair functions. This article discusses the mechanisms by which vanillin enhances or inhibits chemically induced cytotoxicity, mutations, and chromosomal aberrations.

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Non-human toxicity
Rats oral LD50 1580 mg/kg
Rats intraperitoneal LD50 1160 mg/kg
Rats subcutaneous LD50 1500 mg/kg
Mouse intraperitoneal LD50 475 mg/kg
For more complete non-human toxicity data for vanillin (11 in total), please visit the HSDB record page.

Vanillin appears as white or slightly yellow needle-like crystals. It belongs to the benzaldehyde class of compounds, with methoxy and hydroxyl substituents at positions 3 and 4, respectively. It is a plant metabolite with anti-inflammatory, flavoring, antioxidant, and anticonvulsant effects. It belongs to the phenolic, monomethoxybenzene, and benzaldehyde classes of compounds. Vanilla allergen extracts are used in allergen testing. Vanillin has been reported in hops (Humulus lupulus), banyan trees (Ficus erecta var. beecheyana), and several other organisms with relevant data. Vanillin is the main component of vanilla bean extract. Synthetic vanillin sometimes replaces natural vanilla extracts as a flavoring agent in food, beverages, and pharmaceuticals. Like ethyl vanillin, it is widely used in the food industry. Artificial vanilla flavorings are pure vanillin solutions, usually synthesized. Due to the scarcity and high cost of natural vanilla extracts, efforts have long been made to synthesize its main component. The first commercial synthesis of vanillin began with the more readily available natural compound eugenol. Today, synthetic vanillin is made from guaiacol or lignin, a component of wood and a byproduct of the paper industry. (Wikipedia) Vanillin is a metabolite found or produced in Saccharomyces cerevisiae. See also: Vanilla oil (note moved to).

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Therapeutic Uses
The choleretic properties and mechanisms of coumarin and phenolic compounds were explored by studying their effects on parameters such as bile flow, bile acids, electrolytes, and bile metabolites. Vanillin accelerates bile secretion.
Veterinarian: ...as a nebulizer adjunct to help ewes nurse orphaned lambs.
Therapeutic Uses: Methotrexate (MTX) is a chemotherapy drug used to treat cancer that causes cellular genetic damage and exhibits cellular inhibitory effects in various testing systems. Several anti-genotoxic drugs have been studied in various in vitro and in vivo systems. However, data on their ability to modulate MTX-induced genotoxicity are limited. In this study, vanillin (VA) and chlorophyll (CHL) were used as antigenotoxic agents to investigate their ability to reduce MTX-induced DNA damage. V79 Chinese hamster lung cells in the exponential growth phase were treated with five different concentrations (5–100 μg/mL) of MTX for 6 hours, simultaneously activating S9 cells, followed by treatment with two concentrations of VA (50 or 100 μg/mL) or CHL (50 or 100 μg/mL) for 40 hours. Micronucleus (MN) assays were performed with cytochalasin B, and antigenotoxic agents were added to assess micronuclei in binucleated cells. Chromosomal aberrations were assessed in parallel cultures. Results showed that MTX alone dose-dependently reduced the mitotic index (NDI) and mitotic index (MI). A significant increase in the percentage of micronucleated binucleated cells (MNBN) and the percentage of abnormal cells (Abs) was observed. Studies using vanillin (VA) as an antigenotoxic agent have shown that the addition of 50 or 100 μg/mL VA reduced both MNBN numbers (26.3-83.1%) and antibody levels (16.0-87.5%). The addition of chlorhexidine (CHL) also significantly reduced MNBN numbers (53.0-91.5%) at both tested concentrations. Chromosomal aberrations were also significantly reduced (41.0-83.0%). These studies indicate that both VA and CHL can effectively reduce methotrexate (MTX)-induced chromosomal damage. Vanillin is a well-known food and cosmetic additive with antioxidant and antimutagenic properties. Studies have also shown antifungal activity against major human pathogenic fungi, but the effects have been limited. This study investigated the antifungal activity of vanillin and 33 of its derivatives against the human fungal pathogen Cryptococcus neoformans (a major pathogen of cryptococcal meningitis in immunocompromised patients). We found a correlation between the structure of vanillin derivatives and their antifungal activity, indicating that hydroxyl or alkoxy groups in benzaldehyde are more advantageous than halogenated or nitro groups. Among the vanillin derivatives containing hydroxyl or alkoxy groups, o-vanillin and o-ethylvanillin exhibited the highest antifungal activity against Cryptococcus neoformans. We further investigated the antifungal mechanism of o-vanillin. By comparing the transcriptomes of untreated and o-vanillin-treated Cryptococcus neoformans cells using RNA sequencing, we found that this compound can cause mitochondrial dysfunction and induce oxidative stress. These antifungal mechanisms of o-vanillin were experimentally confirmed by mutants lacking mitochondrial function and genes related to oxidative stress response, resulting in significantly reduced growth.

C8H8O3

152.15

152.047

121-33-5

1183

White to off-white solid powder

1.2±0.1 g/cm3

282.6±20.0 °C at 760 mmHg

81-83 °C(lit.)

117.6±15.3 °C

0.0±0.6 mmHg at 25°C

1.588

1.19

1

3

2

11

135

0

MWOOGOJBHIARFG-UHFFFAOYSA-N

InChI=1S/C8H8O3/c1-11-8-4-6(5-9)2-3-7(8)10/h2-5,10H,1H3

4-hydroxy-3-methoxybenzaldehyde

NSC-15351; NSC 15351; Vanillin

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~657.25 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)