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

Endpoint summary

Description of key information

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.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

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

Qualifier:

according to guideline

Guideline:

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

GLP compliance:

not specified

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

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)

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

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

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)

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

Critical effects observed:

not specified

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.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

8 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Data is obtained from K2 source (J check)

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

chronic toxicity: inhalation

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Qualifier:

according to guideline

Guideline:

other:

Principles of method if other than guideline:

Study obtained from Sax's Handbook of Dangerous Industrial Materials and Dictionary of Substances and their effects

GLP compliance:

not specified

Species:

rat

Strain:

not specified

Sex:

not specified

Route of administration:

inhalation

Type of inhalation exposure:

not specified

Vehicle:

not specified

Remarks on MMAD:

MMAD / GSD: no data

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

100 days

Frequency of treatment:

24 hr/day

Remarks:

Doses / Concentrations:
0.3 mg/m³
Basis:
no data

No. of animals per sex per dose:

no data

Control animals:

not specified

Details on results:

Rats showed significant changes in the central nervous system, blood and liver in the correlation of muscle antagonist chronaxy, a reduced number of erythrocytes and hemaglobin, increased methemoglobin level, reticulocytis, leukopenia, hypoproteinemia, a reduced-SH group content in the serum, and accumulation of pyruvic acid in the liver and serum. Dimethylaniline also increased the coproporphyrin urinary excretion and reduced the adrenal ascorbic acid level.

Dose descriptor:

LOAEC

Effect level:

0.3 mg/m³ air

Based on:

test mat.

Sex:

not specified

Basis for effect level:

other: lowest observed advser effect

Critical effects observed:

not specified

Conclusions:

The Lowest observed adverse effect concentration (LOAEC) of N,N-dimethylaniline was found to be 0.3 mg/m³ rats in a 100 day study.
Inhalation of the substance continuously led to anaemia, methaemoglobinaemia, leucopenia and significant pathological changes in the central nervous system.

Executive summary:

Repeated dose inhalation toxicit study was performed using rats for the test compound N, N- dimethylaniline (RA CAS no 121 -69 -7). The Lowest observed adverse effect concentration (LOAEC) of N,N-dimethylaniline was found to be 0.3 mg/m³ rats in a 100 day study. Inhalation of the substance continuously led to anaemia, methaemoglobinaemia, leucopenia and significant pathological changes in the central nervous system.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

LOAEC

0.3 mg/m³

Study duration:

chronic

Species:

rat

Quality of whole database:

K2 level testing data

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Peer reviewed publications for the alternate cas (8004 -87 -3) and the read across chemicals were used to determine the toxic nature of the test compound (CAS no 83968 -28 -9) upon repeated exposure. The studies are summarized below:

Repeated dose toxicity: Oral

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test) was performed (Ministry of Economy, Trade and Industry, Japan, 2016) 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.

In a repeated-dose toxicity study (Abdo et al, 2009), 0, 31.25, 62.5, 125, 250 or 500 mg/kg/day of N,N-dimethylaniline (RA CAS no 121 -69 -7) was administered to 10 male and 10 female Fischer 344 rats and B6C3F1 mice per dose via gavage 5 days/week for 90 days. No mortality was observed. Splenomegaly was observed in all treated groups of rats. Hyperplasia of the bone marrow and hematopoiesis in the spleen were observed in all rats in a dose-related manner. In addition, decreased body weight gain was observed in male rats at 250 and 500 mg/kg/day. Dose-related increases in splenomegaly, and extramedullary hematopoiesis and hemosiderosis of the spleen were observed in the mice when given 31.25 mg/kg/day of N, N-dimethylaniline. Splenomegaly was reported as minimal in 4/10 mice, and extramedullary hematopoiesis and hemosiderois were reported as mild in 1/10 mice.Based observations made, the Low Observed Adverse Effect level (LOAEL) was considered to be 31.25 mg/kg/day in Fischer 344 rats and B6C3F1 mice.

In a repeated-dose toxicity study (USEPA, 2009), Wistar rats (male and female) were administered N,N-diethylaniline via gavage at 0, 10, 50 or 250 mg/kg-bw/day, 7 days/week for 28 days. No mortalities were observed. No changes in body weight, food, and water consumption were reported. Clinical signs of toxicity consisted of increased frequency of respiratory sounds in males at 50 mg/kg-bw/day, and increased frequency of respiratory sounds and salivation in females at 250 mg/kg-bw/day. Hematological effects (decreased red cell counts, decreased hemoglobin concentrations, decreased packed cell volume (PCV) in both sexes and increased mean corpuscular volume (MCV) and mean corpuscular hemoglobin (MCH) in females) were reported at all doses (dose-response not specified). Histological effects were reported for the liver and spleen. In the liver, hemosiderosis of the Kupffer cells at 10 mg/kg-bw/day and extra medullary hematopoeisis at 50 and 250 mg/kg-bw/day were observed. In the spleen, hemosiderosis, extramedullary hematopoiesis and splenic hyperemia were reported at 10 mg/kg-bw/day. Swollen spleens were observed at 50 and 250 mg/kg-bw/day. Increased absolute and relative weights and black pigmentation of the spleen were also reported at 10mg/kg-bw/day. At 50 and 250 mg/kg-bw/day, hyperbilirubinemia, polychromasia were reported, and at 250 mg/kg-bw/day, decreased potassium levels, histopathological findings in the kidneys of both sexes, black pigmentation in the kidneys of females, and increased albumin levels in males were reported. Dose-response and statistical significance were not indicated for any of these observed effects. The Low Observed Adverse Effect level (LOAEL) was considered 10 mg/kg-bw/day (based on hematological and histopathological changes in the spleen and liver consistent with hemolytic anemia).

Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Tes was performed for the test chemical N,N-Dimethylaniline (RA CAS no 121 -69 -7) to evaluate its oral toxic nature upon repeated exposure. The test compund was tested at cocentration of 0, 1, 10 or 100 mg/Kg bw. The male rats were treated for 42 days and females 42 - 46 days (from 14 days before mating to day 4 of lactation). Based upon the observations made, the No Observed Adverse effect level (NOAEL) for the test compound N,N-Dimethylaniline is found to be 1 mg/kg bw in males and < 1 mg/Kg bw in females.

Repeated dose toxicity: Inhalation

Repeated dose inhalation toxicity study (Sax's Handbook of Dangerous Industrial Materials, 2007) was performed using rats for the test compound N, N- dimethylaniline (RA CAS no 121 -69 -7). The Lowest observed adverse effect concentration (LOAEC) of N, N-dimethylaniline was found to be 0.3 mg/m³ rats in a 100 day study. Inhalation of the substance continuously led to anaemia, methaemoglobinaemia, leucopenia and significant pathological changes in the central nervous system.

Repeated dose inhalation toxicity study was performed (Slusar, 1972) using rats for the test compound N, N- dimethylaniline (RA CAS no 121 -69 -7). The Lowest published toxic concentration (TCLo) of N,N-dimethylaniline was found to be 10700 µg/m³ in rats in a 17 week study with intermittent (5 hr) dosage.

 

Based on the data obtained from the alternate CAS no 8004 -87 -3 and the read across data for the target chemical , the substance Formaldehyde, reaction products with N,N dimethylbenzenamine and N-methylbenzenamine, oxidized, hydrochlorides (CAS no 83968 -28 -9) is not likely to show repeated dose toxicity effect by oral and inhalation route and thus cannot be considered for further classification.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:

The No Observed Adverse Effect Level (NOAEL) for the test compound C.I. Basic Violet (8004-87-3) 1 is likely to be 8 mg/Kg bw.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:

The Lowest observed adverse effect concentration (LOAEC) of N, N-dimethylaniline was found to be 0.3 mg/m³ rats in a 100 day study.

Justification for classification or non-classification

Based on the data obtained from the alternate CAS no 8004 -87 -3 and the read across data for the target chemical , the substance Formaldehyde, reaction products with N,N dimethylbenzenamine and N-methylbenzenamine, oxidized, hydrochlorides (CAS no 83968 -28 -9) is not likely to show repeated dose toxicity effect by oral and inhalation route and thus cannot be considered for further classification.