3,3'-Dimethoxybenzidine dihydrochloride is an off-white
powder with a melting point of 274 degrees C.
3,3'-Dimethoxybenzidine is used principally as an intermediate in the production of commercial bisazobiphenyl
dyes for coloring textiles, paper,
plastic, rubber, and leather. In the synthesis of the bisazobiphenyl
dyes, the
amine groups of
3,3'-dimethoxybenzidine are chemically linked with other aromatic
amines. A small quantity of
3,3'-dimethoxybenzidine is also used as an intermediate in the production of
o-dianisidine diisocyanate, which is used in
isocyanate-based adhesive systems and as a component of
polyurethane elastomers.
3,3'-Dimethoxybenzidine dihydrochloride was evaluated in toxicity and carcinogenicity studies as part of the National Toxicology Program's
Benzidine Dye Initiative. This Initiative was designed to evaluate the representative
benzidine congeners and
benzidine congener-derived and
benzidine-derived
dyes.
3,3'-Dimethoxybenzidine dihydrochloride was nominated for study because of the potential for human exposure during production of bisazobiphenyl
dyes and because
benzidine, a structurally related chemical, is a known human
carcinogen. NTP Toxicology and
Carcinogenesis studies were conducted by administering
3,3'-dimethoxybenzidine dihydrochloride (greater than 97.5% pure) in
drinking water to groups of F344/N rats of each sex for 14 days, 13 weeks, 9 months, or 21-months. The 21-month studies were intended to last 24 months but were terminated early because of rapidly declining survival due to
neoplasia. Studies were performed only in rats because similar studies are being performed in mice at the National Center for Toxicology Research. Genetic toxicology studies were conducted with Salmonella typhimurium, Chinese hamster over (CHO) cells, and Drosophila melanogaster. Fourteen-Day Studies: All rats receiving
drinking water concentrations up to 4,500 ppm lived to the end of the studies. Rats that received water containing 4,500 ppm
3,3'-dimethoxybenzidine dihydrochloride lost weight. Water consumption decreased with increasing concentration of chemical and at 4,500 ppm was less than one-fourth that by the controls. Lymphoid depletion of the thymus in males and hypocellularity of the bone marrow in males and females were seen at the 4,500-ppm concentration, but not at the next lower concentration or in controls. Thirteen-Week Studies: All rats receiving concentrations up to 2,500 ppm lived to the end of the studies. Final mean
body weights of rats given
drinking water containing 1,250 or 2,500 ppm
3,3'-dimethoxybenzidine dihydrochloride were 5%-20% lower than those of controls. Water consumption at these concentrations was 40%-60% that consumed by controls. Compound-related effects in rats given water containing 2,500 ppm
3,3'-dimethoxybenzidine dihydrochloride included a mind exacerbation of naturally occurring nephropathy and the presence of a yellow-brown pigment (
lipofuscin) in the cytoplasm of thyroid follicular cells. Serum
triiodothyronine (T3) and
thyroxin (T4) concentrations in females receiving 330 ppm or more and T4 concentrations in males receiving 170 ppm or more were significantly lower than in controls.
Thyrotropin (TSH) concentrations were comparable in controls and exposed rats. Based on the chemical-related nephropathy and reductions in water consumption and
body weight gain observed in the 13-week studies, doses for the long-term studies in male and female rats were 0 or 330 ppm
3,3'-dimethoxybenzidine dihydrochloride in
drinking water administered for 9 months and 0, 80, 170, or 330 ppm administered for 21 months. Nine-Month Studies: Ten rats of each sex in control and 330-ppm groups were evaluated after 9 months. Significant decreases in T3 and T4 concentrations were seen in exposed male and female rats. Other lesions seen in exposed rats included foci of alteration in the liver, a
carcinoma of the preputial gland in one male, a
carcinoma of the clitoral gland in one female, and
carcinoma of the Zymbal gland in two males.
Body Weights and Survival in the Twenty-One-Month Studies: The average amount of
3,3'-dimethoxybenzidine dihydrochloride consumed per day was approximately 6, 12, or 21 mg/kg for low, mid, or high dose male rats and 7, 14, or 23 mg/kg for low, mid, or high dose female rats. Mean
body weights of male and female rats began to decrease relative to those of controls after about 1 year of exposure at 170 or 330 ppm and were 6%-22% lower for males and 7%-17% lower for females. Survival of rats exposed to
3,3'-dimethoxybenzidine dihydrochloride was reduced because animals were dying with
neoplasms or being killed in a moribund condition (survival at 21 months--male: control, 44/60, 73%; low dose, 8/45, 18%; mid dose, 0/75; high dose, 0/60; female: 45/60, 75%; 15/45, 33%; 6/75, 8%; 0/60). Because of these early compound-related deaths, the studies were terminated at 21 months. Nonneoplastic and Neoplastic Effects in the Twenty-One-Month Studies: Increased incidences of several nonneoplastic lesions were observed in exposed rats, including hematopoietic cell proliferation in the spleen and cystic and centrilobular degeneration and
necrosis of the liver.
Neoplasms attributed to
3,3'-dimethoxybenzidine dihydrochloride exposure were observed in rats at many tissue sites, including the skin, Zymbal gland, preputial and clitoral glands, oral cavity, small and large intestines, liver, brain, mesothelium, mammary gland, and uterus/cervix. The incidences of these
neoplasms in male and female rats are given in the abstract summary table (see page 5 of the Technical Report). Genetic Toxicology:
3,3'-Dimethoxybenzidine was mutagenic in S. typhimurium strain TA100 with exogenous metabolic activation and in strain TA98 without activation; a weakly positive response was observed in strain TA1535 with metabolic activation.
3,3'-Dimethoxybenzidine induced sister chromatid exchanges and
chromosomal aberrations in CHO cells with and without exogenous metabolic activation.
3,3'-Dimethoxybenzidine did not induce sex-linked recessive lethal mutations in adult male D. melanogaster exposed via feeding or injection. Conclusions: Under the conditions of these 21-month
drinking water studies, there was clear evidence of carcinogenic activity of
3,3'-dimethoxybenzidine dihydrochloride for male F344/N rats, as indicated by benign and
malignant neoplasms of the skin, Zymbal gland, preputial gland, oral cavity, intestine, liver, and mesothelium. Increased incidences of
astrocytomas of the brain may have been related to chemical administration. There was clear evidence of carcinogenic activity of
3,3'-dimethoxybenzidine dihydrochloride for female F344/N rats, as indicated by benign and
malignant neoplasms of the Zymbal gland, clitoral gland, and mammary gland. Increases in
neoplasms of the skin, oral cavity, large intestine, liver, and uterus/cervix were also considered to be related to chemical administration of
3,3'-dimethoxybenzidine dihydrochloride. Synonyms:
o-dianisidine dihydrochloride; 3,3'-dimethoxy-1,1-
biphenyl)-4,4'-
diamine dihydrochloride; 3,3'-dimethoxy-4,4'-diaminobiphenyl dihydrochloride