Hepatotoxic effects are associated with N,N-Dimethylformamide (DMF; N-Formyldimethylamine)[3].

Absorption, Distribution and Excretion
This solvent can be absorbed through intact skin and also through the lungs. After 4 hours of exposure to 21 ppm of this vapor, the average concentration of dimethylformamide in the blood reached 2.8 μg/L and was undetectable 4 hours after exposure; its metabolite, methylformamide, had an average blood concentration of 1 to 2 mg/L and remained for at least 4 hours after exposure. After 4 hours of exposure to 87 ppm of this vapor, peak blood concentrations of dimethylformamide and methylformamide were observed at 0 and 3 hours, respectively, at approximately 14 μg/L and 8 μg/L. Repeated daily exposure to 21 ppm of dimethylformamide did not result in the accumulation of this chemical or its metabolites in the blood. /Dimethylformamide and Methylformamide/ Eight healthy male subjects were exposed to dimethylformamide (DMF) vapor at a concentration of 8.79 ± 0.33 ppm for 6 hours daily for five consecutive days. All urine samples were collected from subjects from the first exposure to 24 hours after the last exposure, and the methylformamide content in each sample was analyzed. Methylformamide is rapidly excreted from the body, with peak concentrations in urine reaching their peak within hours after each exposure. The mean concentration in samples 7 hours after exposure was 4.74 mg/mL. Only 2-6% of the inhaled dimethylformamide dose is recovered in urine. A significant portion of the absorbed DMF dose is excreted unchanged. The concentration of N-methylformamide in urine is likely the best indicator of the dimethylformamide working index. For more complete data on the absorption, distribution, and excretion of N,N-dimethylformamides (12 in total), please visit the HSDB records page. Metabolism/Metabolites N,N-dimethylformamide (DMF) is primarily metabolized by microsomal cytochrome P-450 to N-hydroxymethyl-N-methylformamide (HMMF), which is further broken down into N-methylformamide (NMF). However, the detailed mechanisms of its toxicity remain unclear. We investigated the metabolism and toxicity of DMF using an ex vivo perfused liver model. DMF was added to the circulating perfusion fluid of ex vivo perfused rat livers at concentrations of 0, 10, and 25 mM. Samples were collected from the inferior vena cava at 0, 30, 45, 60, 75, and 90 minutes after DMF addition. DMF metabolites were analyzed by gas chromatography (GC). Changes in DMF-induced oxygen consumption were monitored during perfusion. Enzyme activities (aspartate aminotransferase: AST, alanine aminotransferase: ALT, and lactate dehydrogenase: LDH) in the perfusion fluid were monitored to observe whether DMF induced hepatotoxicity. As perfusion progressed, the concentration of DMF in the perfusion fluid decreased, while the concentration of NMF increased to a maximum of 1.16 mM. Oxygen consumption increased at DMF concentrations of 10 mM and 25 mM. However, when the perfusion fluid was pretreated with the known cytochrome P-450 inhibitor SKF 525A (300 μM) before the addition of DMF, the oxygen consumption rate was significantly reduced, indicating that the cytochrome P-450 system is responsible for the conversion of DMF to NMF. After the addition of DMF, the activities of AST, ALT, and LDH enzymes increased significantly in a time- and dose-dependent manner. However, the release of these enzymes was inhibited after pretreatment with SKF 525A. …Two groups of researchers studied the metabolism of DMF in volunteers. …Both studies found that most of the absorbed substances were excreted within 24 hours, with N-methylformamide being the main urinary metabolite. Its concentration was related to the exposure intensity. It is known that dimethylformamide is metabolized in the human body through continuous N-demethylation to methylformamide and formamide, and these metabolites are mainly excreted in urine.
Gas chromatography analysis of blood and urine samples from rats and dogs exposed to dimethylformamide (DMF) revealed the presence of N-methylformamide (NMF) and formamide, in addition to DMF. These metabolites were cleared more rapidly in rats than in dogs. Recent studies have shown that the major metabolite of DMF identified by gas chromatography as NMF is not NMF, but rather N-hydroxymethyl-N-methylformamide (HMMF). HMMF is a direct product of the C-hydroxylation of the methyl group in DMF and is a relatively stable methanolamide in aqueous solution. However, it has poor thermal stability and quantitatively decomposes into NMF on a gas chromatography column, potentially further decomposing into formaldehyde. Three studies have confirmed that the metabolite identified as NMF is actually HMMF. One study found a formaldehyde precursor in the urine of mice given DMF. This metabolite only releases formaldehyde upon alkaline hydrolysis. In aqueous solution, pure HMMF also decomposes into formaldehyde only upon alkaline hydrolysis. Another study used high-performance liquid chromatography (HPLC) to separate DMF metabolites in rat urine and then performed mass spectrometry analysis. The observed fragmentation patterns indicated the presence of HMMF, although mass spectrometric fragments, including molecular ions, were also detected in control urine samples. Recently, high-field proton nuclear magnetic resonance (NMR) analysis of urine samples from mice administered DMF provided clear evidence that HMMF, rather than NMF, is the major metabolite of DMF. HMMF exists in two rotational isomers, in which the methyl and formyl protons are not equivalent. In the NMR spectra of the urine, the resonance frequencies corresponding to the methyl and formyl protons in both rotational isomers showed significant signals. However, only a weak signal was observed at the resonance frequency of the methyl proton in NMF. This study found that dimethylamine and methylamine are minor metabolites of DMF in mouse urine. For more complete data on the metabolism/metabolites of N,N-dimethylformamide (14 metabolites in total), please visit the HSDB record page. Dimethylformamide can be absorbed through ingestion, inhalation, and skin contact, and is evenly distributed throughout the body. Its metabolism primarily occurs in the liver via microsomal enzyme systems, producing N-hydroxymethyl-N-methylformamide (DMF-OH) as the main urinary metabolite.

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Biological half-life
Systemic: 4 hours; [TDR, page 551]

Toxicity Summary
Dimethylformamide…is an organic solvent produced in large quantities worldwide. It is used in the chemical industry as a solvent, intermediate, and additive. Dimethylformamide is a colorless liquid with a slightly unpleasant odor, making it poorly detectable; exposure can occur through inhalation of its vapors. Occupational exposure occurs through skin contact with dimethylformamide liquid and vapors. …Inhalation and skin absorption of dimethylformamide can reach toxic doses. Absorbed dimethylformamide is uniformly distributed. Metabolism of dimethylformamide occurs primarily in the liver, aided by microsomal enzyme systems. In animals and humans, the major biotransformation product of dimethylformamide is N-hydroxymethyl-N-methylformamide. This metabolite is converted to N-methylformamide during gas chromatography analysis, while N-methylformamide itself (along with N-hydroxymethylformamide and formamide) is a minor metabolite. In metabolic studies and biomonitoring, urinary concentrations are expressed as N-hydroxymethylformamide. The determination of metabolites in urine may be a suitable bioindicator for total dimethylformamide exposure. In laboratory animals, the metabolism of dimethylformamide has been shown to saturate at high concentrations, and at extremely high concentrations, dimethylformamide inhibits its own metabolism. A metabolic interaction exists between dimethylformamide and ethanol. The environmental effects of dimethylformamide are not fully investigated. Its toxicity to aquatic organisms appears to be low. Dimethylformamide exhibits low acute toxicity in many species. It has mild to moderate skin and eye irritation. A guinea pig study showed that dimethylformamide is not sensitizing. Dimethylformamide can promote the absorption of other chemicals through the skin. Exposure to dimethylformamide in laboratory animals via various routes may lead to dose-related liver injury. Signs of cardiomyopathy and nephropathy have been observed in some studies. Dimethylformamide has not been found to be active in vitro or in vivo, and it has not been found to be active in a wide range of short-term genetic and related effects studies. There are currently no adequate reports on long-term carcinogenicity in laboratory animals. Direct exposure to dimethylformamide in humans has been reported to cause skin irritation and conjunctivitis. Accidental exposure to high concentrations of dimethylformamide can cause abdominal pain, nausea, vomiting, dizziness, and fatigue within 48 hours. Liver function may be impaired, and there have been reports of changes in blood pressure, tachycardia, and abnormal electrocardiograms. Symptoms of prolonged and repeated exposure include headache, loss of appetite, and fatigue. Biochemical markers of liver dysfunction may be observed. Even at concentrations below 30 mg/m³, exposure to dimethylformamide can lead to alcohol intolerance. Symptoms may include sudden facial flushing, chest tightness, and dizziness, sometimes accompanied by nausea and difficulty breathing. …Currently, evidence regarding the carcinogenicity of dimethylformamide in humans is limited. One study reported an increased incidence of testicular tumors, while another showed an increased incidence of oral and pharyngeal tumors, but no increase in testicular tumors. In two studies with limited details, an increased miscarriage rate was reported in women exposed to dimethylformamide and other chemicals. Although the mechanism of action of dimethylformamide is not fully understood, thiocarbamate pesticides have been shown to inhibit aldehyde dehydrogenase. (A2459)

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Toxicity Data
LCLo (rat) = 5,000 ppm/6 hours
LD50: 2800 mg/kg (oral, rat) (T14)
LD50: 1400 mg/kg (intraperitoneal, rat) (T14)
LD50: 3800 mg/kg (subcutaneous, rat) (T14)
LD50: 2000 mg/kg (intravenous, rat) (T14)

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Interactions
In acetone-pretreated male CD1 mice, a single intraperitoneal injection of 1000 mg/kg body weight of dimethylformamide resulted in hepatic necrosis and a significant increase in serum alanine aminotransferase activity. In contrast, no hepatotoxicity was observed in untreated mice receiving the same dose, or in male Sprague Dawley rats receiving either pretreated or untreated doses up to 2000 mg/kg body weight via single intraperitoneal injection. These differences may be related to the significant differences in CYP2EI substrate affinity between rats and mice. In a study of 102 workers, 19 workers experienced symptoms of alcohol intolerance, including facial flushing, sweating, dizziness, and palpitations, primarily occurring within 24 hours of DMF exposure. Of the 34 cases recorded, 26 occurred after workers had consumed alcohol. Of the 13 workers in total, 7 experienced abdominal cramps lasting more than 3 days, 3 had abnormal liver function, and 2 experienced facial flushing. /Dimethylformamide (DMF) Poisoning/ A veterinary euthanasia drug containing embutrothion, mebezoni, tetracaine, and dimethylformamide (DMF; T-61 or Tanax), it can cause severe symptoms and even death after suicidal poisoning. Immediate toxicity is primarily caused by general anesthetics and neuromuscular blocking agents, while delayed hepatotoxicity appears to be related to the solvent DMF. The protective effect of N-acetylcysteine (NAC) remains controversial. Two male veterinarians (aged 50 and 44, respectively) attempted suicide; the former injected T-61 into his chest, while the latter ingested 50 mL orally. Both received NAC treatment (the former for 14 days, the latter for only 20 hours). Urine samples were collected for serial determination of dimethylformamide (DMF), N-methylformamide (NMF), and N-acetyl-S-(N-methylcarbamoyl)cysteine (AMCC). Both patients presented with only mild liver damage. The DMF metabolite NMF appeared rapidly in the urine, while AMCC excretion took longer. The elimination kinetics of DMF and its metabolites were slightly slower than those reported in exposed workers. Although both patients had a good prognosis, there is no clear evidence that NAC directly affects the excretion of NMF and AMCC…
For more complete data on N,N-dimethylformamide interactions (12 items in total), please visit the HSDB records page.

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Non-human toxicity values
Intraperitoneal LD50 in mice: 1120 mg/kg (1.2 mL/kg)
Oral LD50 in mice: 6.8 mL/kg
Intraperitoneal LD50 in Swiss mice: 3.07 g/kg, once daily for 21 days
Intraperitoneal LD50 in tumor-bearing BDF1 mice: 1.23 g/kg, once daily for 9 days
For more complete (30) non-human toxicity values of N,N-dimethylformamide, please visit the HSDB record page.

[1]. Capillary electrophoresis in N,N-dimethylformamide. Electrophoresis. 2005;26(17):3279-3291.

[2]. Solvent artifacts likely to be induced by dimethylformamide. Indian J Physiol Pharmacol. 1986;30(3):248-254.

[3]. Investigation of the mechanistic basis of N,N-dimethylformamide toxicity. Metabolism of N,N-dimethylformamide and its deuterated isotopomers by cytochrome P450 2E1. Chem Res Toxicol. 1993;6(2):197-207.

According to California labor laws and the International Agency for Research on Cancer (IARC) of the World Health Organization, N,N-dimethylformamide is carcinogenic. N,N-Dimethylformamide is a water-white liquid with a slightly fishy odor. Its flash point is 136°F (58°C). Its density is slightly less than water. Its vapor is heavier than air. It is toxic if inhaled or absorbed through the skin. It may irritate the eyes. N,N-Dimethylformamide belongs to the formamide class of compounds, where the amino hydrogen is replaced by a methyl group. It is a polar aprotic solvent, a hepatotoxic substance, and an anti-aging agent. It is a volatile organic compound belonging to the formamide class. Its function is similar to that of formamides. Dimethylformamide is used as an industrial solvent and also in the production of fibers, films, and surface coatings. Acute (short-term) exposure to dimethylformamide in animals and humans has been observed to damage the liver. Symptoms of acute exposure to dimethylformamide in humans include abdominal pain, nausea, vomiting, jaundice, alcohol intolerance, and rash. Long-term occupational inhalation of dimethylformamide can lead to liver damage and digestive disorders in workers. Human studies have suggested a possible link between dimethylformamide (DMF) exposure and testicular cancer, but further research has failed to confirm this association. The U.S. Environmental Protection Agency has not yet classified DMF as a carcinogen. It has been reported that N,N-dimethylformamide is present in tobacco (Nicotiana tabacum) and Cystoseira barbata, with supporting data. N,N-Dimethylformamide (DMF) is a transparent liquid widely used in industry as a solvent, additive, or intermediate due to its good miscibility with water and most common organic solvents. Its health effects include hepatotoxicity and male reproductive toxicity, possibly related to alterations in mitochondrial DNA (mtDNA), including common mtDNA deletions (delta-mtDNA4977) and changes in mtDNA copy number; DMF generates free radicals, including hydroxyl radicals, during biotransformation in vivo. In 2001, global DMF consumption was approximately 285,000 tons, most of which was used as an industrial solvent. Excessive exposure to dimethylformamide (DMF) may lead to hepatotoxicity, alcohol intolerance, potential embryotoxicity and teratogenicity in humans and animals, and decreased sperm motility in humans. Given its widespread use and various toxic effects, DMF has been listed as one of the four compounds prioritized for field human studies by the National Toxicology Program (NTP) of the National Institute of Environmental Health Sciences (NIEHS). Currently, the permissible exposure limit for dimethylformamide (DMF) in the workplace is 10 ppm in both the United States and Taiwan. The American Association of Governmental Industrial Hygienes (ACGIH) recommends using the concentrations of two major DMF metabolites in urine—N-methylformamide (U-NMF) at 15 mg/L and N-acetyl-S-(N-methylcarbamoyl)cysteine (U-AMCC) at 40 mg/L—as a bioexposure indicator (BEI) for workplace DMF exposure (A7735). N,N-Dimethylformamide is a metabolite found or produced in Saccharomyces cerevisiae. It is a formamide in which the amino hydrogen is replaced by a methyl group.

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Mechanism of Action
N,N-Dimethylformamide (DMF) is an organic solvent widely used in the synthetic leather, fiber, and film industries. It can cause hepatotoxicity and carcinogenicity. Although there are numerous experimental and clinical reports on dimethyl fumarate (DMF)-induced liver failure, its toxic mechanism remains unclear. This study aimed to investigate whether the combined use of DMF with low-dose hepatotoxic substances enhances hepatotoxicity and its mechanism of action. Treatment of rats with DMF alone (50-500 mg/kg/day for 3 consecutive days) or a single low-dose carbon tetrachloride (CCl₄, 0.2 mL/kg) resulted in a slight increase in plasma transaminase and lactate dehydrogenase activities. However, the combined use of DMF and CCl₄ significantly exacerbated changes in blood biochemical parameters. Histopathological examination confirmed the synergistic effect of the two in hepatotoxicity. Furthermore, DMF+CCl₄ induces PARP cleavage and caspase-3 activation, but reduces Bcl-xL levels, both of which confirm hepatocyte apoptosis. Consistent with this, the combined administration of DMF+CCl₄ significantly increased lipid peroxidation. In contrast, combined treatment with DMF and lipopolysaccharide, acetaminophen, or D-galactosamine did not enhance hepatotoxicity. Given the link between endoplasmic reticulum (ER) dysfunction and cell death, we monitored ER stress response after DMF and/or CCl₄ treatment. DMF or CCl₄ treatment alone only slightly altered the expression levels of glucose-regulated proteins 78 and 94 and phosphorylated PKR-like ER-localized eIF2α kinase, while combined treatment with DMF and CCl₄ synergistically induced the expression of these proteins and increased the levels of glucose-regulated protein 78 and C/EBP homolog mRNA. These results indicate that combined treatment with DMF and CCl₄ synergistically increases hepatocyte death, which may be related to severe ER stress induction.

C3H7NO

73.09

73.052

68-12-2

6228

Colorless to light yellow liquid

0.948 g/mL at 20 °C

153 °C(lit.)

-61 °C

136 °F

n20/D 1.430(lit.)

0.34

0

1

0

5

33.9

0

CN(C)C=O

ZMXDDKWLCZADIW-UHFFFAOYSA-N

InChI=1S/C3H7NO/c1-4(2)3-5/h3H,1-2H3

N,N-dimethylformamide

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 (1368.18 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.)