53-96-3

Usage

Uses

Used in Research Laboratories:
2-Acetylaminofluorene is used as a laboratory reagent for research purposes, specifically as a positive control for carcinogenicity and mutagenicity studies. It helps researchers understand the mechanisms of cancer and mutation.
Used in Carcinogenicity and Mutagenicity Studies:
2-Acetylaminofluorene is used as a positive control to study the carcinogenicity and mutagenicity of aromatic amines, providing a reference for the identification and quantitation of these substances.
Used in Animal Models for Liver Cancer Research:
2-Acetylaminofluorene is used to induce liver cancer in animal models, such as the 2-AAF/partial hepatectomy rat, to study the mechanism of liver carcinogenesis.
Used in Coal Gasification Processes:
Although it was never marketed due to its carcinogenicity, 2-Acetylaminofluorene is found as a contaminant in coal gasification processes.
Used in Environmental Studies:
The environmental release of 2-Acetylaminofluorene has been monitored and regulated by the US Environmental Protection Agency (EPA) to assess its potential impact on the environment and human health.
Used in Occupational Safety and Health Studies:
2-ACETAMIDOFLUORENE has been studied to understand the risks associated with its exposure, particularly for organic chemists, chemical stockroom workers, and biomedical researchers, who may come in contact with it in the course of their work.
Used in Dose-Response Relationship Studies:
2-Acetylaminofluorene has been used in studies involving large numbers of mice to investigate dose-response relationships, the concept of a threshold level, and the carcinogenic potential of the compound. These studies have provided insights into the factors that influence cancer/tumor development, such as the dose of the carcinogen, the length of exposure, and the gender of the subjects.

Production Methods

2-Acetylaminofluorene is produced for research purposes only with an estimated U.S. annual usage of less than 20 lb. It was originally developed as a possible insecticide but has never been used for this purpose after discovery of its carcinogenicity. It is now almost exclusively used in the laboratory studies as a model carcinogen and mutagen.

Air & Water Reactions

Insoluble in water.

Reactivity Profile

2-ACETAMIDOFLUORENE is incompatible with acids, bases and oxidizing agents. Ozone and chlorinating agents oxidize 2-ACETAMIDOFLUORENE .

Health Hazard

2-Acetylaminofluorene (AAF) is a potent carcinogen in dogs, hamsters, and rats. There is no toxicity information on humans.1

Fire Hazard

Flash point data for 2-ACETAMIDOFLUORENE are not available; however, 2-ACETAMIDOFLUORENE is probably combustible.

Biochem/physiol Actions

A genotoxic carcinogen that is used to model liver carcinogenesis in rat. When N-hydroxylated by cytochrome CYP1A2 in the liver, 2-AAF forms adducts with DNA and is tumorigenic in liver and bladder.

Safety Profile

Confirmed human carcinogen with experimental carcinogenic, neoplas tigenic, tumorigenic, and teratogenic data. Moderately toxic by ingestion and intraperitoneal routes. Experimental reproductive effects. Human mutation data reported. When heated to decomposition it emits toxic fumes of NOx.

Potential Exposure

2-AAF was intended to be used as a pesticide, but it was never marketed because this chemical was found to be carcinogenic. AAF is used frequently by biochemists and technicians engaged in the study of liver enzymes and the carcinogenicity and mutagenicity ofaromatic amines as a positive control. Therefore, these persons may be exposed to AAF.

Carcinogenicity

2-Acetylaminofluorene is reasonably anticipated to be a human carcinogen based on sufficient evidence of carcinogenicity from studies in experimental animals.

Environmental fate

Biological. In the presence of suspended natural populations from unpolluted aquatic systems, the second-order microbial transformation rate constant determined in the laboratory was reported to be 4.8 ± 2.8 x 10-12 L/organism?h (Steen, 1991). Chemical/Physical. Based on first-order rate constants determined at 85.5 °C, hydrolysis halflives at pH values of 2.49, 2.97, 7.34, 9.80, 10.25, and 10.39 were 4.2, 12, 41, 13, 7.2, and 1.9 d, respectively (Ellington et al., 1987). Releases toxic nitrogen oxides when heated to decomposition (Sax and Lewis, 1987).

Shipping

UN3077 Environmentally hazardous substances, solid, n.o.s., Hazard class: 9; Labels: 9-Miscellaneous hazardous material, Technical Name Required.

Purification Methods

Recrystallise it from toluene (1.3mg in 100mL). Its solubility in H2O is 1.3mg/L at 25o, UV: max nm(log ) : 288(4.43), 313(4.13). [Sawicki J Org Chem 21 271 1956.] It can also be recrystallised from 50% AcOH. [Diels et al. Chem Ber 35 3285 1902]. 9-14C and -14C 2-acetamidofluorene were recrystallised from aqueous EtOH and had m 194-195o and 194o respectively. Potent CARCINOGEN. [Miller et al. Cancer Res 9 504 1949, 10 616 1950, Sadin et al. J Am Chem Soc 74 5073 1952, Beilstein 12 H 3287, 12 IV 3373.]

Toxicity evaluation

According to the US EPA’s Toxics Release Inventory, environmental releases of 2-AAF considerably increased from 1998 to 2001, declined to as low as 255 lb in 2003, and have remained below 1000 lb since 2003. However, most of the releases were to hazardous-waste landfills. In 2007, one facility released about 500 lb of 2-AAF to a hazardous-waste landfill and about 250 lb to air. Release of 2-AAF to the environment from artificial sources is probably not significant since less than 20 lb year of 2-ACETAMIDOFLUORENE are consumed in the United States. If released to soil, 2-AAF is expected to have low mobility. Chemical hydrolysis, oxidation, and volatilization are not expected to be significant. If released to water, 2-AAF may undergo direct photolysis and is expected to strongly adsorb to suspended solids and sediments. Chemical hydrolysis, oxidation, volatilization, and bioaccumulation are not expected to be significant. If released to the atmosphere, 2-AAF may undergo vapor phase adsorption to airborne particulate matter, it may react with photochemically generated hydroxyl radicals (estimated vapor phase half-life1/4 5.92 h) or it may undergo direct photolysis.

Incompatibilities

Hygroscopic. Contact with strong oxidizers may cause fire and explosions. Not compatible with cyanides, acids, and/or acid anhydrides. May form unstable and explosive peroxides; a possible polymerization hazard. Contact with strong oxidizers or strong reducing agents may form flammable gases and cause fire and explosions. A weak base that may react as an acid. Incompatible with strong bases (forming potentially dangerous salts), chlorinated hydrocarbons, nitro compounds. Reacts with azo and diazo compounds, generating toxic gases. Contact with mixture of acetic acid 1 dinitrogen trioxide may cause explosion.

Waste Disposal

Consult with environmental regulatory agencies for guidance on acceptable disposal practices. Generators of waste containing this contaminant (≥100 kg/mo) must conform with EPA regulations governing storage, transportation, treatment, and waste disposal. Presumably high-temperature incineration with scrubber for any produced nitrogen oxides can be used

InChI:InChI=1/C15H13NO/c16-15(17)8-10-5-6-14-12(7-10)9-11-3-1-2-4-13(11)14/h1-7H,8-9H2,(H2,16,17)

Upstream product

• 153-78-6 2-aminofluorene 
• 108-24-7 acetic anhydride 
• 64-19-7 acetic acid 
• 6098-44-8 NA-AAF 
• 121-69-7 N,N-dimethyl-aniline 
• 74925-71-6 C₁₅H₁₂NO₅S^(1-)*K⁽¹⁺⁾ 
• 70-18-8 GLUTATHIONE 
• 16808-85-8 N-(sulfonatooxy)-N-acetyl-2-aminofluorene 
• 607-57-8 2-nitro-9H-fluorene 
• 1405-86-3 glycyrrhizin 
• 7647-01-0 hydrogenchloride 
• 110827-07-1 1-fluoren-2-yl-ethanone oxime 
• 10387-40-3 potassium thioacetate 
• 72-89-9 acetylcoenzyme A 

Downstream Products

• 153-78-6 2-aminofluorene 
• 1214-32-0 2-amino-7-nitrofluorene 
• 29005-18-3 3-nitro-fluoren-2-ylamine 
• 43146-78-7 7-Jod-2-acetamino-fluoren 
• 3096-50-2 2-acetylamino-9-fluorenone 
• 92433-40-4 N-(3-chloro-fluoren-2-yl)-acetamide 
• 6942-28-5 N-(3,7-dichloro-fluoren-2-yl)-acetamide 
• 42135-32-0 2-amino-3-nitro-fluoren-9-one 
• 88387-73-9 9-Oxo-2-amino-7-nitrofluorene 
• 72570-99-1 2-acetylamino-7-nitrofluorene 
• 108620-95-7 3-nitro-2-acetamidofluorene 
• 28065-98-7 2-phenylfluorene 
• 479408-24-7 8-(N-2-amino-fluorene)-O⁶-benzyl-3',5'-O-bis(tert-butyldimethylsilyl)-N²-dimethoxytrityl-2'-deoxyguanosine 
• 7151-59-9 9-oxo-2-acetylamino-7-nitrofluorene 

Relevant academic research and scientific papers

Effects of the garlic compounds diallyl sulphide and diallyl disulphide on arylamine N-acetyltransferase activity in Klebsiella pneumoniae

Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene (2-AF) were determined in the bacterium Klebsiella pneumoniae. Cytosols or suspensions of K. pneumoniae with or without specific concentrations of diallyl sulphide (DAS) or diallyl disulphide (DADS) as co-treatment showed different percentages of 2-AF acetylation. The data indicated that there was decreased NAT activity associated with increased levels of DAS or DADS in K. pneumoniae. In growth studies on K. pneumoniae it was demonstrated that DAS or DADS elicited a dose-dependent bacteriocide effect on K. pneumoniae. For the cytosol examinations, the apparent values of K(m) and V(max) were 0.96 ± 0.09 mM and 7.87 ± 0.79 nmol min-1 mg-1 protein, respectively, for 2-AF. However, when DAS or DADS was added to the reaction mixtures, the apparent values of K(m) and V(max) were 0.16 ± 0.04 mM and 0.99 ± 0.16 nmol min-1 mg-1 protein with DAS, respectively, and 0.14 ± 0.18 mM and 0.85 ± 0.10 nmol min-1 mg-1 protein with DADS, respectively, for 2-AF. For the intact bacteria examination, the apparent values of K(m) and V(max) were 0.57 ± 0.06 mM and 2.00 ± 0.14 nmol min-1 per 10 x 1010 CFU, respectively, for 2-AF. However, when DAS or DADS was added to the reaction mixtures, the apparent of values of K(m) and V(max) were 0.41 ± 0.04 mM and 1.30 ± 0.10 nmol min-1 per 10 x 1010 CFU with DAS, respectively, and 0.34 ± 0.04 mM and 1.08 ± 0.08 nmol min-1 per 10 x 1010 CFU with DADS, respectively, for 2-AF. This report is the first demonstration to show that the garlic components DAS and DADS would affect K. pneumoniae growth and NAT activity.

Effects of ibuprofen on arylamine N-acetyltransferase activity in human colon tumor cells

The inhibition of arylamine N-acetyltransferase (NAT) activity by ibuprofen was determined in a human colon tumour (adenocarcinoma) cell line. Two assay systems were employed, one with cellular cytosols (9000 g supernatant) and the other with intact colon tumour cell suspensions. The NAT activity in a human colon tumour cell line was inhibited by ibuprofen in a dose-dependent manner in both systems, i.e. the greater the concentration of ibuprofen in the reaction, the greater the inhibition of NAT activities In both systems. The data also indicated that ibuprofen decreases the apparent K(m) and V(max) of NAT enzyme from human colon tumour cells in both systems examined. This report is the first demonstration to show that ibuprofen affects human colon tumour cell NAT activity.

Effects of the garlic components diallyl sulfide and diallyl disulfide on arylamine N-acetyltransferase activity in human colon tumour cells

Diallyl sulfide (DAS) and diallyl disulfide (DADS), major components of garlic, were used to determine inhibition of arylamine N-acetyltransferase (NAT) activity in a human colon tumour (adenocarcinoma) cell line. Two assay systems were performed, one with cellular cytosols (9000 g suprnatant), the other with intact bacterial cell suspensions. The NAT activity in a human colon tumour cell line was inhibited by DAS and DADS in a dose-dependent manner in both system: that is, the greater the concentration of DAS and DADS in the reaction, the greater the inhibition of NAT activities in both systems. The data also indicated that DAS and DADS decrease the apparent values of K(m) and V(max) of NAT enzymes from human colon tumour cells in both systems examined. This is the first report to demonstrate that garlic components do affect human colon tumour cell NAT activity.

The effects of vitamin E on arylamine N-Acetyltransferase activity in strains of Helicobacter pylori from peptic ulcer patients

Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene (2-AF) and p-aminobenzoic acid (PABA) were determined in the bacterium Helicobacter pylori. Cytosols or suspensions of H. pylori, with or without specific concentrations of vitamin E co-treatment, showed different percentages of 2-AF acetylation. The data indicated that there was increased NAT activity associated with increased levels of vitamin E in H. pylori cytosols and intact bacteria. For the cytosol and intact bacteria examinations, the apparent values of K(m) and V(max) were increased when vitamin E was added to the reaction mixtures for 2-AF and PABA acetylation, respectively. This report is the first demonstration to show that antioxidant agents (vitamin E) can promote H. pylori N-acetyltransferase activity. Copyright (C) 1999 Elsevier Science Ltd.

Ibuprofen affects arylamine N-acetyltransferase activity in Helicobacter pylori from peptic ulcer patients

Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene and p-aminobenzoic acid were determined in the bacterium Helicobacter pylori collected from peptic ulcer patients. Cytosols or suspensions of H. pylori with or without specific concentrations of ibuprofen co-treatment showed different percentages of 2-aminofluorene and p-aminobenzoic acid acetylation. The data indicate that there was decreased NAT activity associated with increased levels of ibuprofen in H. pylori cytosols. Inhibition of growth studies on H. pylori demonstrated that ibuprofen elicited a dose-dependent bactericide effect in H. pylori cultures, i.e. the greater the concentration of ibuprofen, the greater the inhibition of growth to H. pylori. For the cytosol and intact bacteria examinations, the apparent values of K(m) and V(max) were decreased after co-treatment with 40 μM ibuprofen. This report is the first demonstration of ibuprofen inhibition of arylamine N- acetyltransferase activity and ibuprofen inhibition of growth in the bacterium H. pylori.

Rhein affects arylamine N-acetyltransferase activity in Helicobacter pylori from peptic ulcer patients

Arylamine N-acetyltransferase (NAT) activities with 2-aminofluorene and p-aminobenzoic acid were determined in the bacterium Helicobacter pylori collected from peptic ulcer patients. Cytosols or suspensions of H. pylori with or without specific concentrat

One-step reductive amidation of nitro arenes: Application in the synthesis of Acetaminophen

A novel thioacetate mediated one-step reductive acetamidation of aryl nitro compounds was developed and applied to an efficient synthesis of acetaminophen. The reaction also proceeds well without a solvent in the presence of a catalytic amount of surfactant.

Indium-mediated one-pot reductive conversion of nitroarenes to N-arylacetamides

N-Arylacetamides were prepared in excellent yields from nitroarenes in the presence of acetic anhydride, acetic acid and indium by a one-pot procedure.

DNA adducts from nitroreduction of 2,7-dinitrofluorene, a mammary gland carcinogen, catalyzed by rat liver or mammary gland cytosol

Nitrofluorenes are mutagenic and carcinogenic environmental pollutants arising chiefly from combustion of fossil fuels. Nitro aromatic compounds undergo nitroreduction to N-hydroxy arylamines that bind to DNA directly or after O-esterification. This study analyzes the DNA binding and adducts from the in vitro nitroreduction of 2,7-dinitrofluorene (2,7-diNF), a potent mammary carcinogen in the rat. Potential adduct(s) of 2,7-diNF was (were) generated by reduction of 2-nitroso-7-NF with ascorbate/H+ in the presence of calf thymus DNA. The major adduct was characterized by HPLC/ESI/MS and 1H NMR spectrometry as N-(deoxyguanosin-8-yl)-2-amino-7-NF, and a minor one was determined by HPLC/ESI/MS to be a deoxyadenosine adduct of 2-amino-7-NF. Products from enzymatic nitroreduction were monitored by HPLC and DNA adduct formation by 32P-postlabeling. Xanthine oxidase/hypoxanthine-catalyzed nitroreduction of 2,7-diNF, 2-nitrofluorene (2-NF), and 1-nitropyrene (1-NP) yielded the respective amines to similar extents (30-50%). However, the level of the major adducts (～0.15/106 nucleotides) from 2-NF [N-(deoxyguanosin-8-yl)-2-aminofluorene] and 2,7-diNF [N-(deoxyguanosin-8-yl)-2-amino-7-NF] was ≤2% that from 1-NP. In the presence of acetyl CoA, nitroreduction of 2-NF catalyzed by rat liver cytosol/NADH yielded the same adduct at a level of 2.2/106 nucleotides. Liver or mammary gland cytosol with acetyl CoA yielded mainly N-(deoxyguanosin-8-yl)-2-amino-7-NF from 2,7-diNF at >30 adducts/106 nucleotides, levels comparable to those from 1,6-dinitropyrene and 4- or 49-fold greater than the respective levels without acetyl CoA. Recovery of 2-nitroso-7-NF and 2-amino-7-NF from cytosol-catalyzed reduction of 2,7-diNF indicated nitroreduction and an N-hydroxy arylamine intermediate. Likewise, the presence of 2-acetylamino-7-NF indicated that reactivity with acyltransferase(s) was not prevented by the nitro group at C7. These data are consistent with activation of 2,7-diNF via nitroreduction to the N-hydroxy arylamine and acetyl CoA-dependent O-acetylation of the latter to bind to DNA. Enzymatic nitroreduction of 2,7-diNF was greatly enhanced by 9-oxidation. The nitroreduction of either 9-oxo-2,7-diNF or 9-hydroxy-2,7-diNF catalyzed by liver cytosol with acetyl CoA yielded two adducts (>2/106 nucleotides). Differences in the TLC migration of these adducts, compared to those from 2,7-diNF, and the lack of 2,7-diNF formation in the incubations suggested retention of the C9-oxidized groups. The relative ratios of the amine to amide from nitroreductions of 9-oxo-2,7-diNF and 2,7-diNF catalyzed by liver cytosol suggested that the 9-oxo group decreased reactivity with acyltransferase and, thus, the amount of N-acetoxy arylamine that binds to DNA. The mammary gland tumorigenicity of 2,7-diNF and the extent of its activation by the tumor target tissue shown herein suggest relevance of this environmental pollutant for breast cancer.

Synthesis of N,O-Diacetylated N-Arylhydroxylamines by Reduction of Nitroaromatics with Zinc and Acetic Anhydride

Reduction of nitroaromatic compounds with zinc and acetic anhydride in dichloromethane gave N,O-diacetylated N-arylhydroxylamines in good yields under mild conditions.