Formaldehyde, reaction products with N,N-dimethylbenzenamine and N-methylbenzenamine, oxidized, hydrochlorides

Endpoint summary

Description of key information

LD50 was considered to be > 300 mg/kg bw when Crl:CD SD female rats were treated with C.I. Basic Violet 1 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Reference

Endpoint:

acute toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Data is from Ministry of Economy, Trade and Industry, Japan database .

Qualifier:

according to guideline

Guideline:

OECD Guideline 423 (Acute Oral toxicity - Acute Toxic Class Method)

Principles of method if other than guideline:

Single dose acute oral toxicity study of C.I. Basic Violet 1 in female rats

GLP compliance:

not specified

Test type:

acute toxic class method

Limit test:

no

Species:

rat

Strain:

other: Crl:CD (SD)

Sex:

female

Details on test animals or test system and environmental conditions:

Details on test animal
TEST ANIMALS
- Source: No data available.
- Age at study initiation: 9 weeks old
- Weight at study initiation: 1st step : 210.0 (208 ~ 213) g, Step 2: 192.7(189 ~ 195) g, Step 3: 215.3 (214-217)g]
- Fasting period before study: Starved from p.m.5 of the day before dosing to 6 hours after dosing.
- Housing: Individual animal housed in stainless steel cages.
- Diet (e.g. ad libitum): Solid feed [CE-2: CLEA Japan, stock company] ad libitum
- Water (e.g. ad libitum): an automatic water supply was there for animals. The water provided was adapted to the Water Supply Act water quality standards The water supply was ad libitum.
- Acclimation period: No data available.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 21.6-22.5 °C
- Humidity (%): 42 to 48%,
- Air changes (per hr): ventilation frequency 15 times / hour
- Photoperiod (hrs dark / hrs light): Lighting 12 hours/ Day (07:00 to 19:00 on)

Route of administration:

oral: gavage

Vehicle:

other: 0.5%w/v methylcellulose aqueous solution (suspended）

Details on oral exposure:

Details on exposure
VEHICLE
- Concentration in vehicle: 300 and 2000 mg/kg
- Amount of vehicle (if gavage): 10 mL/kg
- Justification for choice of vehicle:
- Lot/batch no. (if required):
- Purity:

MAXIMUM DOSE VOLUME APPLIED:

DOSAGE PREPARATION (if unusual):

CLASS METHOD (if applicable)
- Rationale for the selection of the starting dose: 300 mg/kg (1st), 300 mg/kg (2nd), 2000 mg/kg (3rd)

Doses:

300 and 2000 mg/kg

No. of animals per sex per dose:

Total number of animals 9
300 mg/kg- 3 female rats
2000 mg/kg- 3 female rats

Control animals:

not specified

Details on study design:

Details on study design
- Duration of observation period following administration: 14 days.
- Frequency of observations and weighing:
Body weight was observed immediately before administration, was measured in 2, 5,8,11 and 14 days. Clinical signs were observed daily.
- Necropsy of survivors performed: yes, for 300 and 2000 mg/kg dose group animals.
- Other examinations performed: Mortality, clinical signs, body weight, gross pathology and histopathology.

Statistics:

No data available.

Sex:

female

Dose descriptor:

LD50

Effect level:

> 300 mg/kg bw

Based on:

test mat.

Remarks on result:

other: 50 % mortality not observed

Mortality:

Mortality was observed from day 2 to 9 after treatment. As all the animals in the dose group 2000 mg/kg/day were found mortal within days 2-9 of experiment.

Clinical signs:

other: Clinical sign like Mucus stool, soft stool, Colored stool, colored urine, decrease in stool volume, no stool were observed at dose level of 300 mg/kg/day. Clinical sign like Salivation, mucus stool, soft stool, colored stool (violet), colored urine (viol

Gross pathology:

At dose group of 2000mg/kg/day , Violet discoloration of organs and tissues in all over the body (3/3), dilatation of stomach(3/3), small sizes of spleen (3/3) and thymus(2/3), enlargement of adrenal (2/3), black focus in lung (1/3)were observed.

Other findings:

Histopathology-
At dose group of 2000 mg/kg/day changes observed in the tissue were given below
Stomach: erosion of glandular mucosa(3/3), inflammatory cell infiltration of lamina propria (3/3), hypertrophy of epithelial cell in the glandular stomach (2/3),
Duodenum: erosion of mucosa (3/3)
Intestines: hypertrophy of epithelial cell / inflammatory cell infiltration of lamina propria (3/3), congestion(3/3) /hemorrhage (3/3)/fibrin thrombus of submocosa (1/3)
Liver: enlargement of centrilobular hepatocyte (3/3), necrosis(1/3)/ vacuolation (1/3) of periportal hepatocyte
Adrenal: hypertrophy of zona fasciculata (2/2)
Spleen: Atrophy of follicle/marginal zone /periarterial lymphatic sheath (3/3)
Thymus: Atrophy (2/2) / necrosis of lymphocyte (1/2)
Lung: Congestion (1/1)

Interpretation of results:

Toxicity Category IV

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

LD50 was considered to be > 300 mg/kg bw when Crl:CD SD female rats were treated with C.I. Basic Violet 1 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution.

Executive summary:

In a acute oral toxicity was evaluated in Crl:CD SD female rats by using C.I. Basic Violet 1 in the concentration of 300 and 2000 mg/kg bw as per OECD Guideline 423 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution. Within 2 to 9 days all the animals in 2000 mg/kg bw dose group died with Salivation, Mucus stool, Soft stool, Colored stool (violet), Colored urine (violet), Soiled perineal region, Decrease in stool volume, No stool, Decrease in spontaneous activity, Lacrimation, Emaciation. On 4 day , Decrease in body weight was observed in 300 and 2000 mg/kg bw treated rats. Violet discoloration of organs and tissues in all over the body (3/3), Dilatation of stomach (3/3), Small sizes of spleen (3/3) and thymus (2/3), Enlargement of adrenal (2/3), Black focus in lung (1/3) in gross pathology and Erosion of glandular mucosa (3/3), Inflammatory cell infiltration of lamina propria (3/3), Hypertrophy of epithelial cell in the glandular stomach (2/3) in Stomach, Erosion of mucosa (3/3) in Duodenum, Hypertrophy of epithelial cell / inflammatory cell infiltration of lamina propria (3/3), Congestion (3/3) / hemorrhage (3/3) / fibrinthrombus of submucosa (1/3) in Intestines, Enlargement of cenrtrilobular hepatocyte (3/3), Necrosis (1/3) / vacuolation (1/3) of peri portal hepatocyte in Liver, Hypertrophy of zona fasciculata (2/2) in Adrenal, Atrophy of follicle/marginal zone /periarterial lymphatic sheath (3/3) in Spleen, Atrophy (2/2) / necrosis of lymphocyte (1/2) in Thymus and Congestion (1/1) in Lung were observed in 2000 mg/kg bw treated rats. Therefore, LD50 was considered to be > 300 mg/kg bw when Crl:CD SD female rats were treated with C.I. Basic Violet 1 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution.

Based on the above value the test chemical was classified in category 4 of CLP criteria.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LD50

Value:

300 mg/kg bw

Quality of whole database:

Data is Klimish 2 and from Ministry of Economy, Trade and Industry, Japan database .

Acute toxicity: via inhalation route

Endpoint conclusion

Endpoint conclusion:

no study available

Acute toxicity: via dermal route

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Acute oral toxicity:

Acute oral toxicity of C.I. Basic Violet 1 was summaries as below:

In a study conducted by Ministry of Health, Labour and Welfare, Environment and National Institute of Technology and Evaluation (2007), acute oral toxicity was evaluated in Crl:CD SD female rats by using C.I. Basic Violet 1 in the concentration of 300 and 2000 mg/kg bw as per OECD Guideline 423 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution. Within 2 to 9 days all the animals in 2000 mg/kg bw dose group died with Salivation, Mucus stool, Soft stool, Colored stool (violet), Colored urine (violet), Soiled perineal region, Decrease in stool volume, No stool, Decrease in spontaneous activity, Lacrimation, Emaciation. Decrease in body weight was observed in 300 and 2000 mg/kg bw treated rats. Violet discoloration of organs and tissues in all over the body (3/3), Dilatation of stomach (3/3), Small sizes of spleen (3/3) and thymus (2/3), Enlargement of adrenal (2/3), Black focus in lung (1/3) in gross pathology and Erosion of glandular mucosa (3/3), Inflammatory cell infiltration of lamina propria (3/3), Hypertrophy of epithelial cell in the glandular stomach (2/3) in Stomach, Erosion of mucosa (3/3) in Duodenum, Hypertrophy of epithelial cell / inflammatory cell infiltration of lamina propria (3/3), Congestion (3/3) / hemorrhage (3/3) / fibrinthrombus of submucosa (1/3) in Intestines, Enlargement of cenrtrilobular hepatocyte (3/3), Necrosis (1/3) / vacuolation (1/3) of peri portal hepatocyte in Liver, Hypertrophy of zona fasciculata (2/2) in Adrenal, Atrophy of follicle/marginal zone /periarterial lymphatic sheath (3/3) in Spleen, Atrophy (2/2) / necrosis of lymphocyte (1/2) in Thymus and Congestion (1/1) in Lung were observed in 2000 mg/kg bw treated rats. Therefore, LD50 was considered to be > 300 mg/kg bw when Crl:CD SD female rats were treated with C.I. Basic Violet 1 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution.

In a conducted by Buffalo Cdldr. Cdrp (1982), acute oral toxicity was evaluated in Sprague-Dawley male and female rats by using C.I. Basic Violet 1 in the concentration of 310, 500, 635, 806, 1024 and 1300 mg/kg orally by gavage in corn oil. 50 % mortality was observed at 500, 635, 806, 1024 and 1300 mg/kg. Clinical sign and change in body weight were also observed in treated rats. Gross pathological changes were all observed. Therefore, LD50 was considered to be 413 mg/kg bw when Sprague-Dawley male and female rats were treated with C.I. Basic Violet 1 orally.

In the above similar study, acute oral toxicity was evaluated in Sprague-Dawley male and female rats by using C.I. Basic Violet 1 orally as a 45% solution in 15% acetic acid.50 % mortality was observed at 815 mg/kg. Therefore, LD50 was considered to be 815 mg/kg bw when Sprague-Dawley male and female rats were treated with C.I. Basic Violet 1 orally as a 45% solution in 15% acetic acid.

In a safety assessment report given by Diamante et al (2009) for isomer of Formaldehyde, reaction products with N,N-dimethylbenzenamine and N-methylbenzenamine, oxidized, hydrochlorides, acute oral toxicity was evaluated in 17 young and 12 adult rats by using Basic Violet 1 in the form of 28% of an indelible pencil. Pencil leads were ground into a fine powder, and 3% to 5% lead suspensions in water were used without having been filtered from the undissolved matter. No mortality was observed in both the treated rats. Therefore, LD50 was considered to be > 2160 to 26000 mg/kg when 12 adult rats were treated with Basic violet 1 orally.

Considering the weight of evidence based on the above data for alternate Substance Basic violet 1 (CAS no 8004-87-3), target substance Formaldehyde, reaction products with N,N-dimethylbenzenamine and N-methylbenzenamine, oxidized, hydrochlorides (CAS no 83968-28-9) is likely to be non toxic by oral route. 

Thought, there is a tow non classifying studies are available for the substance C.I. Basic Violet 1, maximum other studies shows LD50 rang from > 300 to 815 mg/kg bw.

Thus, based on weight of evidence, substance C.I. Basic Violet 1 is likely to be classified in Toxicity Category IV as per the CLP regulation.

Justification for selection of acute toxicity – oral endpoint

LD50 was considered to be > 300 mg/kg bw when Crl:CD SD female rats were treated with C.I. Basic Violet 1 orally by gavage in 0.5 w/v% Methylcellulose aqueous solution.

Justification for classification or non-classification

C.I. Basic Violet 1 is likely to be classified in Toxicity Category IV as per the CLP regulation.