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

Repeated dose toxicity: oral

Administrative data

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

combined repeated dose and reproduction / developmental screening

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 J check

Data source

Reference

Reference Type:

other: J check

Title:

Unnamed

Year:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

GLP compliance:

not specified

Test material

Test material information

Constituent 1

Reference substance name:

8004-87-3

EC Number:

616-846-4

Cas Number:

8004-87-3

Molecular formula:

C24H28ClN3

IUPAC Name:

8004-87-3

Constituent 2

Reference substance name:

C.I. Basic Violet 1

IUPAC Name:

C.I. Basic Violet 1

Test material form:

solid: crystalline

Details on test material:

- Name of the test material: C.I. Basic Violet 1
- Molecular weight: 393.95
- Molecular formula: C24H28ClN3
- Appearence: Dark yellow-green to dark green crystal (nub)

Test animals

Species:

rat

Strain:

other: Crl:CD (SD)

Sex:

male/female

Details on test animals or test system and environmental conditions:

- Age at study initiation of dosing: 10 weeks old

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

other: 0.5 % Methylcellulose aqueous solution

Details on oral exposure:

No data

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data

Duration of treatment / exposure:

- Male: 42 days
- Female: 41 - 48 days (from 14 days before mating to day 4 of lactation)

Frequency of treatment:

- Female: 41 - 48 days (from 14 days before mating to day 4 of lactation)

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 1.6, 8, 40 mg/Kg bw (actual ingested)
Basis:

No. of animals per sex per dose:

- Male: 7 rats/group (control and high dose groups of main study) + 5 rats/group (control and high dose groups of recovery).
12 rats/group (low and middle dose groups of main study).
- Female: 12 rats/group (all groups of main study) + 5 rats/group (control and high dose groups of recovery).

Control animals:

yes, concurrent vehicle

Details on study design:

No data

Positive control:

No data

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: No data
- Cage side observations checked in table [No.?] were included. Mortality

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: No data

BODY WEIGHT: Yes
- Time schedule for examinations: No data

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No data

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data
- Time schedule for examinations: No data

OPHTHALMOSCOPIC EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data

HAEMATOLOGY: Yes
- Time schedule for collection of blood: No data
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: No data
- Animals fasted: No data
- How many animals: No data
- Parameters checked in table [No.?] were examined. No data

URINALYSIS: Yes (Males only)
- Time schedule for collection of urine: No data
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked in table [No.?] were examined. No data

NEUROBEHAVIOURAL EXAMINATION: No data
- Time schedule for examinations: No data
- Dose groups that were examined: No data
- Battery of functions tested: sensory activity / grip strength / motor activity / other No data

OTHER: No data

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

No data available

Results and discussion

Results of examinations

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Food efficiency:

not specified

Water consumption and compound intake (if drinking water study):

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, treatment-related

Clinical biochemistry findings:

effects observed, treatment-related

Urinalysis findings:

effects observed, treatment-related

Behaviour (functional findings):

effects observed, treatment-related

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Histopathological findings: neoplastic:

not specified

Details on results:

CLINICAL SIGNS AND MORTALITY
Clinical signs:
1.6 mg/Kg bw: Test article-colored feces
8 mg/Kg bw: Test article-colored feces
40 mg/Kg bw:
Found dead or killed in extremis:
Decreased spontaneous activity, Prone position, Bradypnea, Abnormal respiratory tones, Hypothermia, Abnormal gait, Soft stool, Emaciation, Abdominal distention, Dirty around anus, Soiled perineal region, External genital bleeding and test article-colored feces

Survivals:
Soft stool and dirty around anus (Male/Female),
External genital bleeding (Female)

Mortality:
1.6 mg/Kg bw: Male: 0/12, Female: 0/12
8 mg/Kg bw: Male: 0/12, Female: 0/12
40 mg/Kg bw: Male: 4/12 (day 9, day 11 (3 animals)), Female: 5/12 (day 20 (2 animals), gestational day 6, 18, 21)

BODY WEIGHT AND WEIGHT GAIN: Decrease in the body weight and decrease in the body weight gain (Male), Decrease in the body weight gain (Female) was noted in 40 mg/Kg bw dosed animals

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Decrease in the food consumption was noted in 40 mg/Kg bw dosed animals

FOOD EFFICIENCY: No data available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available

OPHTHALMOSCOPIC EXAMINATION: No data available

HAEMATOLOGY: Increase in the PLT was noted in 40 mg/Kg bw dosed animals

CLINICAL CHEMISTRY:
Killed in extremis:
Increase in the AST and ALT (Male), Increase in the CPK (Male/Female), Increase in the BUN (Male/Female) in 40 mg/Kg bw dosed animals

Survivals:
Decrease in the TP, Increase in the Alb, Decrease in the alpha 1-glb and increase in the A/G (Male), Increase in the BUN (Male), Increase in the BUN (tendency) (Female) in 40 mg/Kg bw dosed animals

URINALYSIS: No adverse effects were noted in any of the dosed grouped animals

NEUROBEHAVIOUR: No data available

ORGAN WEIGHTS: No adverse effects were noted in any of the dosed grouped animals

GROSS PATHOLOGY
1.6 mg/Kg bw: No adverse effects were noted
8 mg/Kg bw: Light violet aqueous content in stomach and cecum (Male 3/12)
40 mg/Kg bw:
Found dead and killed in extremis (Male 4/12, Female 5/12):
Light violet aqueous content and discoloration of mucus membrane in all of the alimentary tract containing oral cavity, subcutaneous tissues and uterus (sporadically noticed in Male and Female), Hydrothorax in thoracic cavity (Male 1/4, Female 2/5), Small thymus (Male 3/4, Female 2/5), Small spleen (Male 1/4, Female 2/5), Edema in thymus (Male 1/4), Reddish urine in gallbladder (Male 1/4), Red discoloration in mucosa of the bladder (Male 1/4), Red discoloration of testis (Male 1/4), Red discoloration of adipose tissue around the testis (Male 1/4), Dilatation of stomach (Female 4/5), Enlargement of adrenal (Female 4/5), Gas retention in stomach (Female 2/5), Dark red viscous retention in vagina (Female 2/5), Dilatation of cecum (Female 1/5), Gas retention in cecum (Female 1/5)

Survivals:
Light violet aqueous content in alimentary tract (Male/Female)

HISTOPATHOLOGY:
40 mg/kg bw:
Found dead or killed in extremis:
Trachea; Desquamation of epithelium and inflammatory cell infiltration of mucosa (Male 1/4, Female 2/5) Glandular stomach; Atrophy of epithelial cell (Male 1/4, Female 1/5) Small/large intestine; Hypertrophy of epithelial cell (sporadically observed in Male and Female) Liver; Hypertrophy of centrilobular hepatocyte (Male 3/4, Female 3/5), Necrosis (Male 1/4, Female 2/5), Vacuolation (Male 1/4) Adrenal; Hypertrophy of zona fasciculata (Male 2/4, Female 5/5) Bone marrow; Deficient erythropoiesis and granulopoiesis (Male 3/4, Female 2/5) Spleen; Atrophy of follicle / marginal zone (Male 2/4, Female 5/5) and periarterial lymphatic sheath (Male 3/4, Female 5/5) Thymus; Atrophy (Male 4/4, Female 3/5) /necrosis of lymphocyte (Male 3/4, Female 4/5) Lymph node; Atrophy of follicle / paracortex (Male 3/4, Female 5/5) Spinal cord / fourth ventricle / testis / urinary bladder; Hemorrhage or hemorrhagic infarction (Male 1/4) Vagina; Hemorrhage (Female 2/5), Mucoid degeneration of mucosa (Female 2/5) Lung; Hemorrhage of alveolus, edema of alveolus and inflammatory cell infiltration (Female 1/5)

Survivals:
Liver; Hypertrophy of centrilobular hepatocyte (Male 2/4) Duodenum; Hypertrophy of epithelial cell (Male 2/4, Female 6/7) Mesenteric lymph node; Sinus histiocytosis (Male 1/4, Female 3/7)

Effect levels

Dose descriptor:

NOAEL

Effect level:

8 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Male/Female 40 mg/kg/day: Death, Hypertrophy of epithelial cell of intestinal tract, Hypertrophy of centrilobular hepatocyte, Necrosis of centrilobular hepatocyte

Target system / organ toxicity

Critical effects observed:

not specified

Applicant's summary and conclusion

Conclusions:

The No Observed Adverse Effect Level (NOAEL) for the test compound C.I. Basic Violet 1 is found to be 8 mg/Kg bw.

Executive summary:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) was performed to determine the toxic nature of the test compound C.I. Basic Violet 1(CAS no 8004 -87 -3) upon repeated exposure to Crl:CD (SD) strain rats. The test chemical was dosed at levels of 0, 1.6, 8 or 40 mg/Kg bw (Actual ingested dose). Male rats were treated for 42 days and female rats were treated from 41 - 48 days (from 14 days before mating to day 4 of lactation). Based on the effects noted at 40 mg/Kg bw, the No Observed Adverse Effect Level (NOAEL) for the test compound C.I. Basic Violet 1 is likely to be 8 mg/Kg bw.