o-Toluidine-d7 (2-Methylaniline-d7; 2-Toluidine-d7; o-Methylaniline-d7)   中文名称: 2-甲苯胺-D7

药物化合物包括碳、氢和其他元素的稳定重同位素，主要作为影响药物开发过程中测量的示踪剂。药物的药代动力学和功能范围可能导致人们对突变的担忧[1]。

Absorption, Distribution and Excretion
We conducted diffusion cell experiments on aniline (ANI), o-toluidine (OT), 4,4'-methylenediphenylamine (MDA), and N-phenyl-2-naphthylamine (PBNA). Human skin was exposed to amino acid solutions of varying concentrations containing water and solvent. Within 24 hours, the recovery rates of ANI in the recipient fluid were approximately 20-38%, and the recovery rate of MDA was 15%. PBNA was not detected in the recipient fluid. We also referenced data from recent studies on OT and β-naphthylamine (BNA). We derived a semi-quantitative ranking of amino acid transdermal absorption as follows: BNA > OT > ANI > MDA > PBNA. For saturated concentrations of ANI in aqueous solution, a linear relationship between exposure dose and permeation was observed. However, the linear extrapolation of pure compound flux, often recommended in risk assessment policies, significantly underestimates transdermal absorption. In vitro data support… the findings of a study of rubber industry workers that transdermal absorption may significantly increase overall AA exposure.
...This study used a diffusion cell to investigate the effects of two skin barrier creams (SBC) and one skin cream (SCC) on the transdermal penetration of aniline (ANI) and o-toluidine (OT), as well as a mixture of OT and workplace-specific lubricants. Experiments were conducted on untreated skin and skin treated with the skin cream. Compared to untreated skin, treated skin showed significantly enhanced transdermal penetration of the test compounds; the enhancement was most significant on skin treated with SBC (water-in-oil emulsion-based) (mean enhancement factor 6.2–12.3 times). Skin treated with SCC showed the lowest enhancement (mean enhancement factor 4.2–9.7 times). In vitro data support...the phenomenon observed in workers where the transdermal absorption of aromatic amines is significantly increased when skin cream is applied. The effectiveness of skin cream in protecting the transdermal penetration of aromatic amines has not yet been confirmed...
...Using a diffusion cell...the penetration of o-toluidine (OT) through human skin was measured. OT is rapid (lag time: approximately 0.8 hours) and has high penetration (50% penetration of the administered dose within 24 hours). Therefore, the use of a moisturizing cream is reasonable…
The distribution of the carcinogen o-toluidine (OT) in rats was studied using diazotization. Three days after the last administration (total treatment duration of 8 months), the level of free amines was 1.5–2 times that of the control group. The highest concentrations were observed in the target tissue gland (Zymbal gland). Furthermore, the concentration of conjugated amines in the gland was also significantly increased (8 times that of the control group). Similar results were reported seven days after the last OT administration: the level of free amines at the same site was 7 times that of the control group. The increased concentrations of free and conjugated amines may indicate their excretion via the Zymbal gland ducts. This may also promote their carcinogenesis.
For more data on the absorption, distribution, and excretion (complete) of 2-aminotoluene (21 in total), please visit the HSDB record page.

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Metabolism/Metabolites
Male F344 rats were subcutaneously injected with 50 or 400 mg/kg of o-[methyl-(14)C]toluidine… The results showed that the main metabolic pathway of o-toluidine in rats was N-acetylation and 4-hydroxylation. Secondary pathways included 6-hydroxylation, methyl oxidation, and amino oxidation. The ratio of sulfate conjugate to glucuronide was 6:1. Substances detected in urine included o-toluidine, azotoluene, o-nitrosotoluene, N-acetyl-o-toluidine, N-acetyl-o-aminobenzyl alcohol, 4-amino-m-cresol, N-acetyl-4-amino-m-cresol, o-aminobenzoic acid, N-acetyl-o-aminobenzoic acid, 2-amino-m-cresol, and unidentified substances. Substances excreted in feces or exhaled air were not mentioned. Four dogs over one year of age received intravenous injection of o-toluidine hydrochloride at a dose of 0.77 mM/kg body weight (equivalent to 111.1 mg/kg body weight), dissolved in water. Blood concentrations of o-toluidine were observed over 6 hours… N-oxidation products were extracted from the blood using carbon tetrachloride… Hemoglobin was estimated by measuring the increase in absorbance at 550 μm after adding cyanide to a blood solution at pH 6.8. The plasma elimination half-life was approximately half an hour. Approximately 10 μg of o-toluidine was detected per milliliter of blood 7 hours after administration. The proportion of hemoglobin to total hemoglobin increased with time after administration, reaching a maximum at 6 hours, indicating the presence of reactive oxidation products. The carbon tetrachloride extract was free of o-nitrosotoluene. Male F344 rats (weighing 230–260 g) were subcutaneously injected with 0.82 mmol/kg body weight o-toluidine (corn oil solution). Urine was collected over 6 hours. All urine samples were analyzed using high-performance liquid chromatography-electrochemical detection within 4 hours of collection. In rats treated with 0.82 mmol/kg body weight of o-toluidine, the concentration of N-hydroxytoluidine in urine ranged from 0.04 to 0.36 μmol from 0 to 6 hours. The activities of metabolic enzymes in the liver, kidneys, and lungs of rats were determined. Male Wistar rats, weighing 200-250 g (n=6 per group), were intraperitoneally injected with 0.75 mg/kg body weight of o-toluidine (dissolved in sunflower oil) for 3 consecutive days. After the last administration, the rats were sacrificed after fasting for 12 hours. On the fourth day, the rats were decapitated, and the liver, kidneys, and lungs were immediately removed, weighed, and homogenized. Analysis was performed (methods not mentioned); a p-value < 0.05 was considered statistically significant. Changes in enzyme activity in various organs: Liver: Cytochrome b5: 0.545 vs 0.447 nmol/mg protein; NADPH cytochrome c reductase: 201.3 vs 165.8 nmol/mg protein/min; Aromatic hydrocarbon hydrolase (AHH): 654 vs 295 pmol/mg protein/min; Glutathione S-transferase to AHH activity ratio: 1926 vs 3969 nmol/mg protein/min; Epoxide hydrolase to AHH activity ratio: 1.85 vs 3.35 nmol/mg protein/min. Kidney: AHH: 70.35 vs 2.91 pmol/mg protein/min; glutathione S-transferase to AHH activity ratio: 2840 vs 63780 nmol/mg protein/min; epoxide hydrolase to AHH activity ratio: 1.24 vs 34.02 nmol/mg protein/min. Lung: AHH: 9.49 vs 4.75 pmol/mg protein/min; glutathione S-transferase to AHH activity ratio: 5184 vs 12484 nmol/mg protein/min; epoxide hydrolase to AHH activity ratio: 4.00 vs 9.89 nmol/mg protein/min.
For more complete data on the metabolisms/metabolites of 2-aminotoluene (14 in total), please visit the HSDB record page.

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Biological Half-Life
In rats (strain not specified), the plasma elimination half-life after oral administration of 500 mg/kg body weight of toluidine was 12 to 15 hours; in dogs, the plasma elimination half-life after intravenous injection of approximately 111 mg/kg body weight was half an hour.

Toxicity Summary
Compound Identification: o-Toluidine is a synthetic chemical, a pale yellow liquid at room temperature. It is primarily used in dye manufacturing, but also in the production of rubber, chemicals, and pesticides, and as a curing agent in epoxy resin systems. Human Exposure: In occupational settings, o-toluidine may exhibit significant carcinogenicity and genotoxicity. Animal Studies: The acute toxicity of o-toluidine is low to moderate, potentially causing mild skin irritation and mild irritation. Information regarding skin or respiratory sensitization by o-toluidine is currently unavailable. The main toxic symptom following short-term acute exposure to this chemical is methemoglobinemia and its associated effects on the spleen. These effects were observed in rats after administration of o-toluidine at a dose of 225 mg/kg body weight/day for 5 consecutive days. The No Observed Adverse Effect Dose (NOEL) has not been determined. In several carcinogenicity studies, oral administration of o-toluidine to rats and mice resulted in a significant increase in the incidence of benign and malignant tumors in various tissues. o-Toluidine is generally not mutagenic in standard bacterial mutagenicity tests, but it is chromosomal breakage-inducing in in vitro mammalian cells. The in vivo genotoxicity of o-toluidine is uncertain; however, some positive results have been reported. Based on the widespread distribution of tumors in animals exposed to o-toluidine and the chromosome breakage activity observed in in vitro mammalian studies, o-toluidine may have genotoxic carcinogenic effects. No information was found related to assessing the risk of reproductive or developmental effects of o-toluidine.

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Toxicity Data
LC50 (rat) = 862 ppm/4hInteractions
Concurrent subcutaneous injection of benzidine and o-toluidine into 116 rats resulted in earlier tumor development. The latency of aniline was twice that of the combined administration (548.6 days vs. 264.1 days, respectively).

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Non-human toxicity values
Oral LD50 in rats: 940 mg/kg
Oral LD50 in rats: 670 mg/kg
Wistar oral LD50 in rats: 0.75 mL/kg body weight (750 mg/kg body weight)
Intraperitoneal LD50 in male Sprague-Dawley rats: 164 mg/kg body weight
For more complete non-human toxicity data for 2-aminotoluene (out of 15), please visit the HSDB records page.

[1]. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019 Feb;53(2):211-216.

According to an independent committee of scientific and health experts, o-toluidine is potentially carcinogenic. o-Toluidine appears as a colorless or pale yellow transparent liquid. It may turn reddish-brown upon exposure to air and light. Its flash point is 85 degrees Celsius (185 degrees Fahrenheit). Its density is similar to water, and it is slightly soluble in water. Its vapor is heavier than air. It has been identified as a carcinogen. o-Toluidine is an aminotoluene, in which the amino substituent is located ortho-to the methyl group. It is a carcinogen. o-Toluidine is primarily used in dye manufacturing. o-Toluidine is highly toxic to humans through skin absorption, inhalation of vapors, or ingestion. Acute (short-term) exposure to o-Toluidine can affect the blood (e.g., methemoglobinemia) and cause clinical symptoms of central nervous system depression. Long-term exposure to o-Toluidine can cause chronic (long-term) effects such as anemia, anorexia, weight loss, skin lesions, central nervous system depression, cyanosis, and methemoglobinemia. Animal studies have shown that long-term exposure to o-toluidine can damage the spleen, liver, bladder, and blood. Occupational exposure to dyes (including o-toluidine) is associated with an increased risk of bladder cancer. 2-Methylaniline hydrochloride (the hydrochloride salt of o-toluidine) is carcinogenic to rats and mice. O-toluidine has been classified as a Group 2 carcinogen by the U.S. Environmental Protection Agency (EPA), meaning it is likely carcinogenic to humans. O-toluidine has been reported to be present in tea (Camellia sinensis), and relevant data exist. O-toluidine is a synthetic, light-colored, light-sensitive liquid, slightly soluble in water, and miscible with carbon tetrachloride, ether, and ethanol. O-toluidine hydrochloride is a synthetic, light-sensitive white crystalline powder, soluble in dimethyl sulfoxide and ethanol. O-toluidine and its hydrochloride are primarily used as intermediates in the production of dyes and pigments. O-toluidine decomposes upon heating, releasing toxic nitrogen oxide fumes, while the hydrochloride salt also produces hydrochloric acid. Four studies on workers exposed to o-toluidine reported an increased incidence of bladder cancer. o-Toluidine and its hydrochloride are likely human carcinogens. (NCI05)
Toluidine's chemical properties are very similar to aniline and share properties with other aromatic amines. Due to the amino group attached to the aromatic ring, toluidine is weakly basic. All toluidine compounds have low solubility in pure water, but will dissolve in acidic solutions due to the formation of ammonium salts, as is typical for organic amines. At room temperature and pressure, o-toluidine and m-toluidine are viscous liquids, while p-toluidine is a flaky solid. This can be explained by the more symmetrical molecule of p-toluidine, making it easier to form a crystal structure. p-Toluidine can be prepared by reducing p-nitrotoluene. p-Toluidine reacts with formaldehyde to form the Trog base.

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Mechanism of Action
The mechanism of DNA damage caused by the carcinogen o-toluidine metabolites in the presence of metals was studied using DNA sequencing technology with 32P-labeled human DNA fragments. The main metabolite, 4-amino-3-methylphenol, causes DNA damage in the presence of Cu(II). The primary DNA cleavage sites are thymine and cytosine residues. o-Nitrotoluene is a minor metabolite that does not induce DNA damage even in the presence of Cu(II), but the addition of NADH induces DNA damage very effectively. Its DNA cleavage pattern is similar to that of 4-amino-3-methylphenol. Bartophenonelin and catalase inhibit DNA damage induced by these o-toluidine metabolites, indicating that Cu(I) and H₂O₂ are involved in the DNA damage process. Typical hydroxyl radical scavengers have no inhibitory effect on DNA damage. In the presence of Cu(II), o-toluidine metabolites increase the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine in calf thymus DNA. UV-Vis and electron spin resonance (ESR) spectroscopy studies show that 4-amino-3-methylphenol undergoes auto-oxidation in the presence or absence of Cu(II) to generate aminomethylphenoxy radicals, while o-nitrosotoluene is reduced to o-toluenehydronitrooxy radicals under the action of NADH. Therefore, these free radicals are thought to react with O₂ to generate O₂⁻, which in turn generates H₂O₂, and the reactive species produced by the reaction of H₂O₂ with Cu(I) are involved in DNA damage. The metal-dependent DNA damage mediated by H₂O₂ via o-toluidine metabolites appears to be associated with the carcinogenicity of o-toluidine. This study investigated the in vivo covalent binding of o-toluidine and p-toluidine to rat liver macromolecules to determine whether there was a correlation between the degree of binding of each isomer and its carcinogenic potency. The ortho isomer has been shown to have stronger hepatotoxicity than the para isomer. In addition to macromolecular binding, the tissue distribution of each isomer was measured. The degree of binding of both isomers to liver macromolecules appeared to reach its maximum 24–28 hours post-administration. At 24 hours post-administration, the binding level of o-toluidine to DNA was approximately 1.2 times lower than that of p-toluidine. The binding of o-toluidine to RNA and proteins was also lower than that of p-toluidine, but the difference was not as significant as the difference in DNA binding. The tissue distribution of the two isomers showed subtle differences. However, contrary to the macromolecular binding data, the area under the plasma concentration-time curve for o-toluene was approximately 1.8 times greater than that for p-toluene. Based on the results of these studies, there is no direct correlation between the degree of macromolecular binding and carcinogenic potency.

C7H2D7N

114.20

114.117

68408-22-0

97917-08-3;636-21-5 (hydrochloride)

7242

Colorless to light yellow liquid

2.7 °F (NTP, 1992)
-14.41 °C
Melting point: -23.7 °C (alpha form); -14.7 °C (beta form)
-16.3 °C
-16 °C (beta-form)
2.7 °F
6 °F

2.158

26.02

1

1

0

8

70.8

0

[2H]C1=C([2H])C([2H])=C(N)C(C([2H])([2H])[2H])=C1[2H]

RNVCVTLRINQCPJ-UHFFFAOYSA-N

InChI=1S/C7H9N/c1-6-4-2-3-5-7(6)8/h2-5H,8H2,1H3

2-methylaniline

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

注意: 如下所列的是一些常用的体内动物实验溶解配方，主要用于溶解难溶或不溶于水的产品（水溶度<1 mg/mL）。 建议您先取少量样品进行尝试，如该配方可行，再根据实验需求增加样品量。

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注射用配方1: DMSO : Tween 80： Saline = 10 : 5 : 85 (如： 100 μL DMSO → 50 μL Tween 80 → 850 μL Saline)
*生理盐水/Saline的制备：将0.9g氯化钠/NaCl溶解在100 mL ddH ₂ O中，得到澄清溶液。
注射用配方 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL DMSO → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)
注射用配方 3: DMSO : Corn oil = 10 : 90 (如： 100 μL DMSO → 900 μL Corn oil)
示例: 以注射用配方 3 (DMSO : Corn oil = 10 : 90) 为例说明, 如果要配制 1 mL 2.5 mg/mL的工作液, 您可以取 100 μL 25 mg/mL 澄清的 DMSO 储备液，加到 900 μL Corn oil/玉米油中, 混合均匀。

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注射用配方 4: DMSO : 20% SBE-β-CD in Saline = 10 : 90 [如：100 μL DMSO → 900 μL (20% SBE-β-CD in Saline)]
*20% SBE-β-CD in Saline的制备（4°C，储存1周）：将2g SBE-β-CD (磺丁基-β-环糊精) 溶解于10mL生理盐水中，得到澄清溶液。
注射用配方 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (如： 500 μL 2-Hydroxypropyl-β-cyclodextrin (羟丙基环胡精)→ 500 μL Saline)
注射用配方 6: DMSO : PEG300 : Castor oil : Saline = 5 : 10 : 20 : 65 (如： 50 μL DMSO → 100 μL PEG300 → 200 μL Castor oil → 650 μL Saline)
注射用配方 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (如： 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
注射用配方 8: 溶解于Cremophor/Ethanol (50 : 50), 然后用生理盐水稀释。
注射用配方 9: EtOH : Corn oil = 10 : 90 (如： 100 μL EtOH → 900 μL Corn oil)
注射用配方 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (如： 100 μL EtOH → 400 μL PEG300 → 50 μL Tween 80 → 450 μL Saline)

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口服配方 1: 悬浮于0.5% CMC Na (羧甲基纤维素钠)
口服配方 2: 悬浮于0.5% Carboxymethyl cellulose (羧甲基纤维素)
示例: 以口服配方 1 (悬浮于 0.5% CMC Na)为例说明, 如果要配制 100 mL 2.5 mg/mL 的工作液, 您可以先取0.5g CMC Na并将其溶解于100mL ddH2O中，得到0.5%CMC-Na澄清溶液；然后将250 mg待测化合物加到100 mL前述 0.5%CMC Na溶液中，得到悬浮液。

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口服配方 3: 溶解于 PEG400 (聚乙二醇400)
口服配方 4: 悬浮于0.2% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 5: 溶解于0.25% Tween 80 and 0.5% Carboxymethyl cellulose (羧甲基纤维素)
口服配方 6: 做成粉末与食物混合

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注意: 以上为较为常见方法，仅供参考， InvivoChem并未独立验证这些配方的准确性。具体溶剂的选择首先应参照文献已报道溶解方法、配方或剂型，对于某些尚未有文献报道溶解方法的化合物，需通过前期实验来确定（建议先取少量样品进行尝试），包括产品的溶解情况、梯度设置、动物的耐受性等。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。