Reaction mass of Methanaminium, N-[4-[[4-(dimethylamino)phenyl]phenylmethylene]-2,5-cyclohexadien-1-ylidene]-N-methyl- & acetate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Description of key information

Reproductive toxicity study

Data available for the read across chemicals was reviewed to determine the reproductive toxicity of Reaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate.Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 30-40mg/kg bw . Thus, comparing this value with the criteria of CLP regulationReaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate is likely to classify as reproductive toxicant.

Link to relevant study records

Reference

Endpoint:

screening for reproductive / developmental toxicity

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Remarks:

Experimental data of read across substances

Justification for type of information:

Data for the target chemical is summarized based on the structurally similar read across chemicals

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

WoE report is based on two reproductive toxicity studies on rats 1.The three-generation reproductive toxicity study of test material was performed in Fischer 344 rats.2.Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test in Rats

GLP compliance:

not specified

Limit test:

no

Justification for study design:

No data available

Specific details on test material used for the study:

- Name of test material: Reaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate- Molecular formula: C25H28N2O2- Molecular weight: 388.508 g/mole- Smiles : [N+](=C1/C=CC(\C=C1)=C(\c1ccccc1)c1ccc(cc1)N(C)C)(\C)C.C(C)(=O)[O-]- Inchl: 1S/C23H25N2.C2H4O2/c1-24(2)21-14-10-19(11-15-21)23(18-8-6-5-7-9-18)20-12-16-22(17-13-20)25(3)4;1-2(3)4/h5-17H,1-4H3;1H3,(H,3,4)/q+1;/p-1- Substance type: Organic- Physical state: maroon lustrous liquid

Species:

rat

Strain:

other: 1.Fischer 344 ,2.Crj: CD(SD)

Details on species / strain selection:

No data available

Sex:

male/female

Details on test animals or test system and environmental conditions:

2.- Age at study initiation of dosing: 10 weeks old

Route of administration:

oral: feed

Vehicle:

other: Feed

Details on exposure:

2.PREPARATION OF DOSING SOLUTIONS: Dose were prepared by suspendingtest substance in 0.5 % Methylcellulose aqueous solution at 0, 1.6, 8 and 40 mg/kg bw.

Details on mating procedure:

1.- M/F ratio per cage:No data available - Length of cohabitation: 14 days - Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:No data available - After ... days of unsuccessful pairing replacement of first male by another male with proven fertility.No data available - Further matings after two unsuccessful attempts: [no / yes (explain)]No data available - After successful mating each pregnant female was caged (how): single - Any other deviations from standard protocol:No data available

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

No data available

Duration of treatment / exposure:

Study 1total:280-294F0 generation :at least 80days F1b generation:100-140 days F2b generation:100-140 days F3a generation:up to weaning Study 2Male: 42 day Female: 41 - 48 days (from 14 days before mating to day 4 of lactation)

Frequency of treatment:

Daily

Details on study schedule:

Study 1- F1 parental animals not mated until [...] weeks after selected from the F1 litters.After at lest 80 daysfrom start of study, In F0 generation male and female rats of same dose group were caged together for 14 days for mating .Pups from this mating F1a generation used for separate study while F0 generation animals were mating second times in same dose group following birth of F1b generation - Selection of parents from F1 generation when pups were [...] days of age.: At 100-140 days of age, one male and one female from each litter were randomly selected to produce F2a generation. Same procedure was used to produce F2b generation - Age at mating of the mated animals in the study: [...] weeksAfter 100-140 days, F3a generation were produce as result of mating one male and one female animal per litter from F2b generation.

Remarks:

Study 10, 5,15 and 30 mg /kg bw /dayStudy 20,1.6,8.0,40.0 mg/kg bw /day

No. of animals per sex per dose:

Study 1Total :450Control group:180Male :90 female 90treated group:2705mg/kg bw/day:45male and 45 female 15mg/kg bw/day:45male and 45 female30mg/kg bw/day:45male and 45 femaleStudy 2Total: 1060 mg/kg/day: 7 male, 12 female 1.6 mg/kg/day: 12 male, 12 female8 mg/kg/day: 12 male, 12 female40 mg/kg/day: 7 male, 12 femaleRecory group:0 mg/kg/day: 5 male, 5 female 40 mg/kg/day: 5 male, 5 female

Control animals:

yes, concurrent vehicle

Details on study design:

No data available

Positive control:

No data available

Parental animals: Observations and examinations:

Study 1CAGE SIDE OBSERVATIONS: No data - Time schedule: - Cage side observations checked in table [No.?] were included. DETAILED CLINICAL OBSERVATIONS: No data - Time schedule: BODY WEIGHT: Yes - Time schedule for examinations: Study 2.Survival, clinical sign, body weight, food consumption and urineanalysis, FOB, Urinalysis (Male only), were examined.

Oestrous cyclicity (parental animals):

Estrous cyclicity, copulation and implantation were examined.

Sperm parameters (parental animals):

No data available

Litter observations:

Study 1STANDARDISATION OF LITTERS- Performed on day 4 postpartum: [yes/no]: No data available - If yes, maximum of [...] pups/litter ([...]/sex/litter as nearly as possible); for F1b and F2a generation one male and one female from each litter were seleted and 2 male and 2 female per litter for F3a generation were selected excess pups were killed and discarded. PARAMETERS EXAMINEDThe following parameters were examined in [F1 / F2 / F3] offspring:yes [number and sex of pups, stillbirths, live births, postnatal mortality, presence of gross anomalies, were observed GROSS EXAMINATION OF DEAD PUPS:[no / yes, for external and internal abnormalities; possible cause of death was/was not determined for pups born or found dead.]No data available ASSESSMENT OF DEVELOPMENTAL NEUROTOXICITY:No data available ASSESSMENT OF DEVELOPMENTAL IMMUNOTOXICITY:No data available Study 2.Number of pups, number of live pups, sex ratio on days 0 and 4, clinical signs and body weight were examined.

Postmortem examinations (parental animals):

Study 1SACRIFICE - Male animals: All surviving animals [describe when, e.g. as soon as possible after the last litters in each generation were produced.]No data available - Maternal animals: All surviving animals [describe when, e.g. after the last litter of each generation was weaned.] No data available GROSS NECROPSY - Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]No data available HISTOPATHOLOGY / ORGAN WEIGHTS The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively.: yes in F3a generations were observed Study 2.Hematology, clinical chemistry, Organ weight, gross pathology and histopathology were examined.

Postmortem examinations (offspring):

Study 1SACRIFICE - The F1 offspring not selected as parental animals and all F2 offspring were sacrificed at [#?] days of age. - These animals were subjected to postmortem examinations (macroscopic and/or microscopic examination) as follows: No data available GROSS NECROPSY - Gross necropsy consisted of [external and internal examinations including the cervical, thoracic, and abdominal viscera.]No data available HISTOPATHOLOGY / ORGAN WEIGTHS The tissues indicated in Table [#] were prepared for microscopic examination and weighed, respectively. yes in F3a generation Study 2.Gross pathology were examined.

Statistics:

No data available

Reproductive indices:

2.Fertility , gestation period, implantation index, live birth index, delivery index were examined.

Offspring viability indices:

Viability index on 0 and 4 day were examined.

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Study 2. When treated with 40 mg/kg/day, 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 were observed in dead treated rats as compared to control. Soft stool and dirty around anus in male and female and External genital bleeding in Female survival rats were observed. Colored feces were observed in 1.6 and 8 mg/kg bw treated rats.

Dermal irritation (if dermal study):

not specified

Mortality:

mortality observed, non-treatment-related

Description (incidence):

Study 1: No dose related effect on mortality was observed Study 2.When treated with 40 mg/kg bw, 1 male and 4 female died and 3 male and 1 female were sacrificed due to morbidity. No effect on survival of treated rats were observed at 1.6 and 8 mg/kg bw as compared to control.

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Study 1:The dose related effect in body weight was observed at dose 30mg/kg bw /day group compared to control and lower dose group.Study 2.When treated with 40 mg/kg/day, decrease in body-weight and body-weight gain were observed in male and body weight gain in female rats as compared to control

Food consumption and compound intake (if feeding study):

effects observed, treatment-related

Description (incidence and severity):

Study 2.When treated with 40 mg/kg/day, decrease in food consumption was observed in male and female rats as compared to control.

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

Description (incidence and severity):

Study 2.When treated with 40 mg/kg/day, Increase in the PLT was observed in male and female rats as compared to control.

Clinical biochemistry findings:

effects observed, treatment-related

Description (incidence and severity):

Study 2.When treated with 40 mg/kg/day, Increase in the AST and ALT in male, CPK and BUN were observed in extremely killed male and female rat. Decrease in the TP and alpha 1-glb, Increase in the Alb, A/G and BUN in male and Increase in the BUN (tendency) in Female rats were observed as compared to control in survival animals.

Urinalysis findings:

not specified

Behaviour (functional findings):

effects observed, treatment-related

Description (incidence and severity):

Study 2.When treated with 40 mg/kg/day, Bradypnea, Prone position, Decrease in spontaneous activity, Incomplete eyelid opening and abnormal gait in died animals and no effect on surviving animals were observed.

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Study 2.When treated with 40 mg/kg/day, 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) were observed as compared to control in extremis killed animals. Liver; Hypertrophy of centrilobular hepatocyte (Male 2/4), Duodenum; Hypertrophy of epithelial cell (Male 2/4, Female 6/7) and Mesenteric lymph node and Sinus histiocytosis (Male 1/4, Female 3/7) were observed as compared to control in survival animals.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

no effects observed

Description (incidence and severity):

Study 2. No effect on Estrous cyclicity, copulation and inplantation were observed.

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

Study 1: No effect observed on fertility index and stillborn animals ,sex ratio, number of pups per litter .Study 2.No effect on Reproductive performance of treated male and femlae rats were observed as compared to control.

Dose descriptor:

NOAEL

Effect level:

> 30 - <= 40 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other: No effect observed on reproductive performance

Critical effects observed:

not specified

System:

other: not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Description (incidence):

Study 1;not dose related mortality was observed,

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

Study 1: The dose related effect in body weight was observed at dose 30mg/kg bw /day group compared to control and lower dose group

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

Description (incidence and severity):

Study 1: No effect observed on fertility index and stillborn animals ,sex ratio, number of pups per litter .

Dose descriptor:

NOAEL

Effect level:

30 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

reproductive performance

Remarks on result:

other: No effect observed on reproductive performace

Critical effects observed:

not specified

System:

other: not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Description (incidence and severity):

Study 2.No Clinical signs were observed in treated pups as compared to control.

Dermal irritation (if dermal study):

not specified

Mortality / viability:

no mortality observed

Description (incidence and severity):

Study 2.No effect on suvival of pups were observed as compared to control.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Study 2.No effect on body weight of pups were observed as compared to control.

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

Study 1:No dose related effects on incidence of gross deformities were noted Study 2.No gross pathological changes were observed in treated pups as compared to control.

Histopathological findings:

not specified

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

NOAEL

Generation:

other: F2b

Effect level:

30 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: gross deformities

Remarks on result:

other: No effect observed at dose 30mg/kg bw

Dose descriptor:

NOAEL

Generation:

F1

Effect level:

40 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

not specified

Basis for effect level:

viability

clinical signs

mortality

body weight and weight gain

gross pathology

Remarks on result:

other: No effects on developmental parameters

Critical effects observed:

not specified

System:

other: not specified

Treatment related:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality / viability:

not specified

Body weight and weight changes:

not specified

Food consumption and compound intake (if feeding study):

not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Sexual maturation:

not specified

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

effects observed, treatment-related

Description (incidence and severity):

Study 1:Dose related trend for focal dilatation of the renal cortex and tubules ,for necrosis of the thymus and inverse dose-response relationship for red pulp haematopoietic cell proliferation of the spleen

Other effects:

not specified

Behaviour (functional findings):

not specified

Developmental immunotoxicity:

not specified

Dose descriptor:

LOAEL

Generation:

other: F3a

Effect level:

5 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

histopathology: non-neoplastic

Remarks on result:

other: Dose related effect observed

Critical effects observed:

yes

Lowest effective dose / conc.:

5 mg/kg bw/day (nominal)

Organ:

kidney

spleen

thymus

Treatment related:

yes

Dose response relationship:

yes

Relevant for humans:

not specified

Reproductive effects observed:

not specified

Treatment related:

not specified

Relation to other toxic effects:

not specified

Dose response relationship:

not specified

Relevant for humans:

not specified

Conclusions:

NOAEL for reproductive toxicity was considered to be above 30 -40mg/kg bw/day for P, P1 and F2b generation when male and female rats were treated with Reaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate orally.

Executive summary:

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of Reaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate .The studies are as mentioned below:

Study 1.

In three generation reproductive toxicity study, Total 450 male and female Fischer 344 rats were used in the study separated in two group i.e. control group and treated group.90 animals of each sex were used in control group. Test material was given in feed at dose concentration 0, 5, 15 and 30 mg /kg bw /day.45 animals of each sex were used for each dose concentration in F0 generation.

 

In F0 generation male and female rats of same dose group were caged together for 14 days for mating .Pups from this mating F_1ageneration used for separate study while F0 generation animals were mating second times in same dose group following birth of F_1bgeneration. At 100-140 days of age, one male and one female from each litter were randomly selected to produce F_2ageneration. Same procedure was used to produce F_2bgeneration .After 100-140 days, F3a generation were produce as result of mating one male and one female animal per litter from F2b generation. Pups from all generations were examined for gross deformities while at weaning, 2male and 2 female per litter of F3a generation were randomly selected for histopathology of organs and tissues.

 

The dose related effect in body weight was observed at dose 30mg/kg bw /day dose group, as animals in this group had lower body weight compared to control and lower dose group. No dose related effects were observed on reproductive performance as number of pups per litter, fertility index and numbers of stillborn animals were not affected across the generation or dose group.Also No dose related effects were observed on gross deformities in each generation.The only significant histopathological changes noted in F_3ageneration were dose related trend for focal dilation of renal cortex and tubules and statistically significant dose related trend for necrosis of thymus (p<0.012 for male and p<0.0001 for female).Also statistically significant inverse dose response relationship for red pulp hematopoietic cell proliferation of spleen (p<0.001 for male and p<0.00001 for female).Hence, NOAEL for reproductive toxicity was considered to be 30mg/kg bw/day for P, P1 and F2b generation and LOAEL w as considered to be 5mg/kg bw/day for F3a generation when male and female Fischer 344 rats were treated with test material orally.

 

Study 2.

In a Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test,Crl:CD (SD)male and female rats treated with test material in the concentration of 0, 1.6, 8 and 40 mg/kg/day orally by gavage in0.5 % Methylcellulose aqueous solution. 1 male and 4 female died and 3 male and 1 female were sacrificed due to morbidity at 40 mg/kg bw. No effect on survival of treated rats were observed at 1.6 and 8 mg/kg bw as compared to control, 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 were observed in dead treated rats as compared to control. Soft stool and dirty around anus in male and female and External genital bleeding in Female surviving rats at 40 mg/kg/day. Colored feces were observed in 1.6 and 8 mg/kg bw treated rats.Decrease in body-weight and body-weight gain were observed in male and body weight gain in female rats and decrease in food consumption were observed in male and female rats as compared to control at40 mg/kg/day.Bradypnea, Prone position, Decrease in spontaneous activity,incompleteeyelid opening and abnormal gait in died animals and noeffects on surviving animals wereobserved.No effect on ureinanalysis wasobserved in male rats as compared to control.Similarly,Increase in the PLT was observed in male and female ratsas compared to control at40 mg/kg/day. Increase in the AST and ALT in male, CPK and BUN were observed in extremely killed male and female rat at 40 mg/kg/day. Decrease in the TP and alpha 1-glb, Increase in the Alb, A/G and BUN in male and increase in the BUN (tendency) in Female rats were observed as compared to control in survival animals. No effect on organ weight of treated was male and female rats were observed as compared to control. 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) observed in extremely killed animals and Light violet aqueous content in alimentary tract in surviving male and female rats at 40 mg/kg/day. Light violet aqueous content in stomach and cecum were observed in male rats (3/12) at 8 mg/kg/day. 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) were observed as compared to control in extremis killed animals and Liver; Hypertrophy of centrilobular hepatocyte (Male 2/4), Duodenum; Hypertrophy of epithelial cell (Male 2/4, Female 6/7), Mesenteric lymph node and Sinus histiocytosis (Male 1/4, Female 3/7) were observed as compared to control in survival animals at 40 mg/kg/day. In addition no reproductive effect such as Estrous cyclicity, copulation and implantation, Fertility rat, gestation period, implantation index, live birth index, delivery index were observed in treated rats as compared to control. No effect on viability, development and growth of pups were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 40 mg/kg/day for P and F1 generation whenCrl:CD (SD)male and female rats treated with test material orally by gavage.

 

 

 

Effect on fertility: via oral route

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

30 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

Data is Klimicsh 2 and from authoritative database

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

Reproductive toxicity study

Data available from different studies were reviewed to determine the reproductive toxicity of Reaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate .The studies are as mentioned below:

Study 1.

In three generation reproductive toxicity study, Total 450 male and female Fischer 344 rats were used in the study separated in two group i.e. control group and treated group.90 animals of each sex were used in control group. Test material was given in feed at dose concentration 0, 5, 15 and 30 mg /kg bw /day.45 animals of each sex were used for each dose concentration in F0 generation.

 

In F0 generation male and female rats of same dose group were caged together for 14 days for mating .Pups from this mating F_1ageneration used for separate study while F0 generation animals were mating second times in same dose group following birth of F_1bgeneration. At 100-140 days of age, one male and one female from each litter were randomly selected to produce F_2ageneration. Same procedure was used to produce F_2bgeneration .After 100-140 days, F3a generation were produce as result of mating one male and one female animal per litter from F2b generation. Pups from all generations were examined for gross deformities while at weaning, 2male and 2 female per litter of F3a generation were randomly selected for histopathology of organs and tissues.

 

The dose related effect in body weight was observed at dose 30mg/kg bw /day dose group, as animals in this group had lower body weight compared to control and lower dose group. No dose related effects were observed on reproductive performance as number of pups per litter, fertility index and numbers of stillborn animals were not affected across the generation or dose group.Also No dose related effects were observed on gross deformities in each generation.The only significant histopathological changes noted in F_3ageneration were dose related trend for focal dilation of renal cortex and tubules and statistically significant dose related trend for necrosis of thymus (p<0.012 for male and p<0.0001 for female).Also statistically significant inverse dose response relationship for red pulp hematopoietic cell proliferation of spleen (p<0.001 for male and p<0.00001 for female).Hence, NOAEL for reproductive toxicity was considered to be 30mg/kg bw/day for P, P1 and F2b generation and LOAEL w as considered to be 5mg/kg bw/day for F3a generation when male and female Fischer 344 rats were treated with test material orally.

 

Study 2.

In a Combined Repeated Dose Toxicity Study with the Reproduction/ Developmental Toxicity Screening Test,Crl:CD (SD)male and female rats treated with test material in the concentration of 0, 1.6, 8 and 40 mg/kg/day orally by gavage in0.5 % Methylcellulose aqueous solution. 1 male and 4 female died and 3 male and 1 female were sacrificed due to morbidity at 40 mg/kg bw. No effect on survival of treated rats were observed at 1.6 and 8 mg/kg bw as compared to control, 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 were observed in dead treated rats as compared to control. Soft stool and dirty around anus in male and female and External genital bleeding in Female surviving rats at 40 mg/kg/day. Colored feces were observed in 1.6 and 8 mg/kg bw treated rats.Decrease in body-weight and body-weight gain were observed in male and body weight gain in female rats and decrease in food consumption were observed in male and female rats as compared to control at40 mg/kg/day.Bradypnea, Prone position, Decrease in spontaneous activity,incompleteeyelid opening and abnormal gait in died animals and noeffects on surviving animals wereobserved.No effect on ureinanalysis wasobserved in male rats as compared to control.Similarly,Increase in the PLT was observed in male and female ratsas compared to control at40 mg/kg/day. Increase in the AST and ALT in male, CPK and BUN were observed in extremely killed male and female rat at 40 mg/kg/day. Decrease in the TP and alpha 1-glb, Increase in the Alb, A/G and BUN in male and increase in the BUN (tendency) in Female rats were observed as compared to control in survival animals. No effect on organ weight of treated was male and female rats were observed as compared to control. 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) observed in extremely killed animals and Light violet aqueous content in alimentary tract in surviving male and female rats at 40 mg/kg/day. Light violet aqueous content in stomach and cecum were observed in male rats (3/12) at 8 mg/kg/day. 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) were observed as compared to control in extremis killed animals and Liver; Hypertrophy of centrilobular hepatocyte (Male 2/4), Duodenum; Hypertrophy of epithelial cell (Male 2/4, Female 6/7), Mesenteric lymph node and Sinus histiocytosis (Male 1/4, Female 3/7) were observed as compared to control in survival animals at 40 mg/kg/day. In addition no reproductive effect such as Estrous cyclicity, copulation and implantation, Fertility rat, gestation period, implantation index, live birth index, delivery index were observed in treated rats as compared to control. No effect on viability, development and growth of pups were observed in treated rats as compared to control. Therefore, NOAEL was considered to be 40 mg/kg/day for P and F1 generation whenCrl:CD (SD)male and female rats treated with test material orally by gavage.

 

 

Based on the data available for the read across chemical, No Observed Adverse Effect Level (NOAEL) was considered to be above 30-40mg/kg bw . Thus, comparing this value with the criteria of CLP regulationReaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetateis likely to classify as reproductive toxicant.

 

Effects on developmental toxicity

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Justification for classification or non-classification

Thus, comparing this value with the criteria of CLP regulationReaction mass of Methanaminium, N-​[4-​[[4-​(dimethylamino)​phenyl]​phenylmethylene]​-​2,​5-​cyclohexadien-​1-​ylidene]​-​N-​methyl-​ & acetate is likely to classify as reproductive toxicant.