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Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Administrative data

Description of key information

Repeated Dose toxicity (Oral)
The summaries of the various studies presented as weight of evidence having Klimisch rating 2 are presented in the table below:
S. No End pointValue (mg/kg bw)SpeciesKlimisch Rating
1.NOAEL7142.85Mouse2
2.NOAEL500Dog2
3.NOAEL2500Rat2
4.NOAEL2857Mouse2
Thus, it can be seen that the no observed adverse effect level (NOAEL) values in the various chronic studies ranges from 500 to 7142.85 mg/kg bw in various animal species tested. Hence, it can be concluded that the chemical dihydrogen (ethyl)[4-[4-[ethyl(3-sulphonatobenzyl)amino](4-hydroxy-2-sulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, disodium salt (Synonym Fast Green FCF/ FD&C Green No. 3) is not likely to exhibit toxic effects, within the dose levels mentioned upon chronic repeated dosing by the oral route.
Repeated Dose (Dermal)
In a Chronic dermal toxicy Study, Swiss Webster male and female mice treated with F D & C Green No. 3, the concentration of 0 and 1.0 % twice weekly. The results shows that F D & C Green NO. 3.was not toxic dermally. No effect was observed on survival and clinical signs of treated mice. High or low body weights observed late in treated group were due to only a few surviving animals which reduced the biological significance of the averages. In addition, No significant incidence of gross changes was observed in treated mice. Malignant lymphoma of liver, Kidneys and Lungs, Myeloid metaplasia of Spleen were observed in treated mice. There was no indication of treatment related changes since most lesions were also evident in the control.
  
Therefore, NOAEL was considered to be 1.0 % (1428.5 mg/kg) when Swiss Webster male and female mice were treated with FD & C Green NO. 3.by dermal application for 19.5 months.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic 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 from WHO Food Additive Series
Qualifier:
according to guideline
Guideline:
other: Repeated dose carcinogenicity
Principles of method if other than guideline:
Repeated dose carcinogenicity study of FD&C Green No. 3 orally in mice
GLP compliance:
not specified
Limit test:
no
Species:
mouse
Strain:
other: Charles-River CD-1
Sex:
male/female
Details on test animals or test system and environmental conditions:
- Age at study initiation: 43 days study
Route of administration:
oral: feed
Vehicle:
other: diet
Details on oral exposure:
mice were fed diets containing 0, 0.5, 1.5, or 5.0% Fast Green FCF
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
24 months
Frequency of treatment:
Daily
Remarks:
Doses / Concentrations:0, 0.5, 1.5 and 5.0 % (714.3, 2143 and 7142.85 mg/kg/day respectively)Basis:nominal in diet
No. of animals per sex per dose:
Total : 3000 %: 60 male, 60 female 0.5 % 30 male, 30 female 1.5 % : 30 male, 30 female 5.01 % : 30 male, 30 female
Control animals:
yes, concurrent vehicle
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes - Time schedule: No data available- Cage side observations checked in table [No.?] were included: Mortality and morbidity was observedDETAILED CLINICAL OBSERVATIONS: No data available - Time schedule: No data availableBODY WEIGHT: Yes - Time schedule for examinations: No data available FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): No data available- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data available- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data available FOOD EFFICIENCY: No data available- 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 availableWATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No data available - Time schedule for examinations: No data available OPHTHALMOSCOPIC EXAMINATION: No data available - Time schedule for examinations: No data available- Dose groups that were examined: No data available HAEMATOLOGY: Yes - Time schedule for collection of blood: At 3, 6, 12, 18 months and on completion of study.- Anaesthetic used for blood collection: No data available - Animals fasted: No data available - How many animals: 10 animals from each group at 3, 6, 12, 18 months and all on completion of study. - Parameters checked in table [No.?] were examined: Haemoglobin, haematocrit, and erythrocyte counts were examined. CLINICAL CHEMISTRY: No data available- Time schedule for collection of blood:- Animals fasted: No data available- How many animals: No data available- Parameters checked in table [No.?] were examined: No data available URINALYSIS: No data available- Time schedule for collection of urine: No data available- Metabolism cages used for collection of urine: No data available - Animals fasted: No data available- Parameters checked in table [No.?] were examined: No data available NEUROBEHAVIOURAL EXAMINATION: No data available - Time schedule for examinations: No data available- Dose groups that were examined: No data available- Battery of functions tested: sensory activity / grip strength / motor activity / other: No data availableOTHER: No data available
Sacrifice and pathology:
GROSS PATHOLOGY: Yes Gross changes/tissue masses were examined for all animals in treated dose groups. HISTOPATHOLOGY: Yes Tissues were examined histologically from all control and 5% dose groups as well as all animals dying or killed in extremis from these groups Organ examined:Adrenals, aorta, bone and marrow (femur), brain (3 sections), eyes (with optic nerve), gall bladder, gastrointestinal tract (oesophagus, stomach,duodenum, ileum, caecum, colon), heart, kidneys, liver, lung, lymph nodes (mesenteric and mediastinal), mammary gland, nerve (sciatic), ovaries, pancreas, pituitary, prostate, salivary gland, seminal vesicles, skeletal muscle, skin, spinal cord, spleen, testes with epididymides, thymus, thyroid/parathyroid, trachea, urinary bladder, uterus and gross lesions/tissue masses were examined.
Clinical signs:
no effects observed
Description (incidence and severity):
No mertality were observed in treated male and female mice as compare to control.
Mortality:
no mortality observed
Description (incidence):
No mertality were observed in treated male and female mice as compare to control.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
When treated with 5 % in male mice mean body weight was significantly decreased at weeks 52 and 78 as compare to control. In female mice mean body weight was consistently decreased as compare 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:
effects observed, treatment-related
Description (incidence and severity):
When treated with 5% in male mice slight reductions in haemoglobin, haematocrit, and erythrocyte counts were observed at 18 months but no other consistent or dose-related haematological changes were observed.
Clinical biochemistry findings:
not specified
Urinalysis findings:
not specified
Behaviour (functional findings):
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Details on results:
-Clinical signs and mortalityMortality : No mertality were observed in treated male and female mice as compare to control.Clinical sign: No data available -Body weight and weight gain: When treated with 5 % in male mice mean body weight was significantly decreased at weeks 52 and 78 as compare to control.In female mice mean body weight was consistently decreased as compare to control.-Haematology:When treated with 5% in male mice slight reductions in haemoglobin, haematocrit, and erythrocyte counts were observed at 18 months but no other consistent or dose-related haematological changes were observed.-Histopathology: Histological examination did not reveal any treatment-related lesions and the incidence, origins and histology of benign and malignant neoplasms did not differ significantly between controls and treated animals.
Dose descriptor:
NOAEL
Effect level:
7 142.85 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No adverse effect on survival, body weight, hematology, gross pathology and histopathology
Critical effects observed:
not specified
Conclusions:
NOAEL for repeated dose toxicity study was considered to be 5% (7142.85 mg/kg/day) in male and female mice when exposed to FD&C Green No. 3 by oral route for 24 months. 
Executive summary:

A Chronic study was conducted to evaluate the toxic effects of repeated administration of FD&C Green No. 3 to male and female mice by feed. FD&C Green No. 3 was administered to mice in diet at dosages of 0, 0.5, 1.5 and 5.0 % (714.3, 2143 and 7142.85 mg/kg/day) for 24 months. No mortalities occurred that could be directly attributed to treatment. Change was observed in body weight of male and female mice at 5 % but no other consistent differences in body weight were noted. Similarly, no changes were observed in hematology of female mice, in male mice change was observed in haemoglobin, haematocrit, and erythrocyte counts at 18 months but no other consistent or dose-related haematological changes were observed. In addition, no change were observed in histopathology of treated male and female mice when compare with control. Therefore, NOAEL for repeated dose toxicity study was considered to be 5% (7142.85 mg/kg/day) in male and female mice when exposed toFD&C Green No. 3by oral route for 24 months. 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
7 142.85 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
weight of evidence approach from studies published in peer reviewed journals

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Data is from peer reviewed journal
Qualifier:
according to guideline
Guideline:
other: Skin painting studies
Principles of method if other than guideline:
Skin painting studies in Swiss Webster mice were carried out.
GLP compliance:
not specified
Limit test:
yes
Species:
mouse
Strain:
Swiss Webster
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS- Source:- Age at study initiation:- Weight at study initiation: 17 to 25 g- Fasting period before study: No data available- Housing: All groups were equally divided as to sex. Mice of the same sex were housed five per cage.- Diet (e.g. ad libitum): Purina Laboratory Chow ad libitum- Water (e.g. ad libitum): Fresh- Acclimation period: No data availableENVIRONMENTAL CONDITIONS- Temperature (°C): No data available- Humidity (%):No data available- Air changes (per hr): No data available- Photoperiod (hrs dark / hrs light): No data availableIN-LIFE DATES: From: To: No data available
Type of coverage:
not specified
Vehicle:
water
Remarks:
Distilled water
Details on exposure:
TEST SITE- Area of exposure: dorsal area- % coverage: 6 cm2- Type of wrap if used: No data available- Time intervals for shavings or clipplings: according to the rate of hair growth.REMOVAL OF TEST SUBSTANCE- Washing (if done): No data available- Time after start of exposure: No data availableTEST MATERIAL- Amount(s) applied (volume or weight with unit): No data available- Concentration (if solution):0.1 ml - Constant volume or concentration used: yes- For solids, paste formed: Not applicableVEHICLE- Justification for use and choice of vehicle (if other than water): Distilled water was used- Amount(s) applied (volume or weight with unit): No data available- Concentration (if solution): No data available- Lot/batch no. (if required): No data available- Purity: No data availableUSE OF RESTRAINERS FOR PREVENTING INGESTION: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
19.5 months
Frequency of treatment:
Twice weekly
Remarks:
Doses / Concentrations:1.0 % (1428.5 mg/kg)Basis:no data
No. of animals per sex per dose:
Total: 300 0 % : 100 male, 100 female1.0 % : 50 male, 50 female
Control animals:
yes, concurrent vehicle
Details on study design:
50 mice/ sex/dose were used and 0.1 ml of the test material was applied to the dorsal of each mouse with rubber applicator.Animals that died, those sacrificed moribund and those surviving the 19.5 month experimental period were necropsied and tissues were preserved in 10% formalin.
Positive control:
No data available
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes - Time schedule: Daily - Cage side observations checked in table [No.?] were included: Observations were done to record mortality DETAILED CLINICAL OBSERVATIONS: Yes - Time schedule: Daily DERMAL IRRITATION (if dermal study): No data BODY WEIGHT: Yes, Time schedule for examinations: Daily FOOD CONSUMPTION:- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No data availableFOOD 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 availableWATER CONSUMPTION: No data OPHTHALMOSCOPIC EXAMINATION: No data availableHAEMATOLOGY: No data availableCLINICAL CHEMISTRY: No data available URINALYSIS: No data availableNEUROBEHAVIOURAL EXAMINATION: No data availableOTHER: No data available
Sacrifice and pathology:
GROSS PATHOLOGY: Yes Grossly abnormal organs or tissues were observed. HISTOPATHOLOGY: YesOrgans were fixed in 10% formalin solution.Tissues examined.Brain, Pituitary, Thyroid, Thymus, Liver, Spleen, Kidney, Adrenal, Stomach, Small intestines, Large intestines, Urinary bladder, Axillary lymph node, Kidney, ovary, Skin from area of treatment, Any tissue masses and grossly abnormal organs or tissues were examined.
Other examinations:
No data available
Statistics:
No data available
Clinical signs:
no effects observed
Description (incidence and severity):
No treatment related effect were observed on survival of mice as compared to control. No effect was observed on behavior of treated mice as compared to control.
Dermal irritation:
not specified
Mortality:
no mortality observed
Description (incidence):
No treatment related effect were observed on survival of mice as compared to control. No effect was observed on behavior of treated mice as compared to control.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
High or low body weights observed late in treated group were due to only a few surviving animals which reduced the biological significance of the averages.
Food consumption and compound intake (if feeding study):
not examined
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
not examined
Haematological findings:
not examined
Clinical biochemistry findings:
not examined
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
not examined
Gross pathological findings:
no effects observed
Description (incidence and severity):
No significant incidence of gross changes was observed in treated mice as compared to control.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Malignant lymphoma of liver, Kidneys and Lungs, Myeloid metaplasia of Spleen were observed in treated mice. There was no indication of treatment related changes since most lesions were also evident in the controls.
Histopathological findings: neoplastic:
not specified
Details on results:
The findings described appear to indicate freedom from significant systemic and/or dermal toxic manifestations associated with administrations.
Dose descriptor:
NOAEL
Effect level:
1 428.5 mg/kg bw/day
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: Survival, Clinical signs, Body weight, Gross pathology and histopathology
Critical effects observed:
not specified
Conclusions:
NOAEL was considered to be 1.0 % (1428.5 mg/kg) when Swiss Webster male and female mice were treated with F D & C Green NO. 3.
Executive summary:
In a Chronic dermal toxicy Study, Swiss Webster male and female mice treated with F D & C Green No. 3, the concentration of 0 and 1.0 % twice weekly. The results shows that F D & C Green NO. 3.was not toxic dermally. No effect was observed on survival and clinical signs of treated mice. High or low body weights observed late in treated group were due to only a few surviving animals which reduced the biological significance of the averages. In addition, No significant incidence of gross changes was observed in treated mice. Malignant lymphoma of liver, Kidneys and Lungs, Myeloid metaplasia of Spleen were observed in treated mice. There was no indication of treatment related changes since most lesions were also evident in the control.

 

Therefore, NOAEL was considered to be 1.0 % (1428.5 mg/kg) when Swiss Webster male and female mice were treated with FD & C Green NO. 3.by dermal application for 19.5 months.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 428.5 mg/kg bw/day
Study duration:
chronic
Species:
mouse
Quality of whole database:
Reliable with restriction

Additional information

Repeated dose toxicity (Oral)

The summaries of the various studies presented as weight of evidence having Klimisch rating 2 are presented in the table below:

S. No

End point

Value (mg/kg bw)

Species

Klimisch Rating

1.       

NOAEL

7142.85

Mouse

2

2.       

NOAEL

500

Dog

2

3.       

NOAEL

2500

Rat

2

4.       

NOAEL

2857

Mouse

2

 

Thus, it can be seen that the no observed adverse effect level (NOAEL) values in the various chronic studies ranges from 500 to 7142.85 mg/kg bw in various animal species tested. Hence, it can be concluded that the chemical dihydrogen (ethyl)[4-[4-[ethyl(3-sulphonatobenzyl)amino](4-hydroxy-2-sulphonatobenzhydrylidene]cyclohexa-2,5-dien-1-ylidene](3-sulphonatobenzyl)ammonium, disodium salt (Synonym Fast Green FCF/ FD&C Green No. 3) is not likely to exhibit toxic effects, within the dose levels mentioned upon chronic repeated dosing by the oral route.

Repeated Dose Toxicity (Dermal)

In a Chronic dermal toxicy Study, Swiss Webster male and female mice treated withF D & C Green No. 3, the concentration of 0 and 1.0 % twice weekly. The results shows that F D & C Green NO. 3.was not toxic dermally. No effect was observed on survival and clinical signs of treated mice. High or low body weights observed late in treated group were due to only a few surviving animals which reduced the biological significance of the averages. In addition, No significant incidence of gross changes was observed in treated mice. Malignant lymphoma of liver, Kidneys and Lungs, Myeloid metaplasia of Spleen were observed in treated mice. There was no indication of treatment related changes since most lesions were also evident in the control.

 

Therefore, NOAEL was considered to be 1.0 % (1428.5 mg/kg) when Swiss Webster male and female mice were treated with FD & C Green NO. 3.by dermal application for 19.5 months.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Data is from an authoritative source and is part of weight of evidence studies.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
This end point was considered for waiver since repeated exposure of humans via inhalation to the chemical FD&C Green is highly unlikely taking into account the extremely low vapour pressure of 1.7E-43 mm Hg at 25 deg C. In addition, the possibility of exposure to inhalable particles is also negligible given the particle size distribution of the chemical which have larger particle size than which can be inhaled. Thus, repeated exposure by the inhlation route for this chemical appears highly unlikely.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
data is from peer reviewed journal

Justification for classification or non-classification

From the available data on chronic repeated dose studies of the chemical FD&C Green No. 3, by the oral and dermal route of exposure, it can be concluded that based upon the high no observed adverse effect level (NOAEL) values, the chemical is not likely to exhibit toxic effects on repeated exposure within the dose levels mentioned in the studies. Exposure by the inhalation route for this chemical is also unlikely given the low vapour presure of the chemical.

Thus, the chemical is not likely to be classified for repeated dose toxicity by the oral, inhalation and dermal route of exposure.