Disodium 6-hydroxy-5-[(4-sulphonatophenyl)azo]naphthalene-2-sulphonate

Endpoint summary

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records

Reference

Endpoint:

two-generation reproductive toxicity

Remarks:

based on test type (migrated information)

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Principles of method if other than guideline:

The reproductive effects of Sunset Yellow FCF on male and female Charles River CD rats by oral (Diet) route at different dose levels was examined.

GLP compliance:

not specified

Species:

mouse

Strain:

other: Crj: CD-1

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Japan Inc., Kanagawa, Japan
- Age at study initiation: four weeks of age
- Weight at study initiation: Not available
- Fasting period before study: Not available
- Housing: housed individually in polycarbonate solid-floored cages with wood flakes
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C):24 +/-1 deg c
- Humidity (%):55+/-5 deg c
- Air changes (per hr): Not available
- Photoperiod (hrs dark / hrs light): 12-h light/dark cycle

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: basal diet(Nihon Clea, CE-2)

Details on exposure:

DIET PREPARATION
- Mixing appropriate amounts with (Type of food):Nihon Clea, CE-2

Details on mating procedure:

- M/F ratio per cage: 1:1
- Length of cohabitation: 5 days
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy: no data
- Further mating after two unsuccessful attempts: [no / yes (explain)] Not available
- After successful mating each pregnant female was caged (how): individually

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

17 weeks

Frequency of treatment:

Daily

Details on study schedule:

At nine weeks of age, each female was paired with one male from the same treatment group for a period of five days. The males were removed from the females after five days, and the females were allowed to carry their litters to term, and deliver and rear all of their offspring.
In the F 1 generation, litter size, litter weight, and sex ratio (male/female) were measured on postnatal day (PND) 0 (at birth). The offspring were weighed individually on PNDs 0, 4, 7,14, and 21 during the lactation period. The offspring were weaned when they were four weeks old, and one male and one female were selected randomly to continue treatment from each litter. The animals were weighed individually at four, five, six, seven, eight, and nine weeks of age after weaning.

Remarks:

Doses / Concentrations:
0,300,600,1200 mg/kg bw
Basis:

No. of animals per sex per dose:

0mg/kg bw (Control):10 female and 10 male mice
300 mg/kg bw (low dose group):10 female and 10 male mice
600 mg/kg bw (mid dose group):10 female and 10 male mice
1200 mg/kg bw (high dose group):10 female and 10 male mice

Control animals:

yes, concurrent vehicle

Parental animals: Observations and examinations:

DETAILED CLINICAL OBSERVATIONS: Yes
BODY WEIGHT: Yes
- Time schedule for examinations: weighed individually on experimental days 0, 2, 4, 7, 14, 21, 28, and 30 during the preconception period.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
OTHER:
Movement activity of exploratory behavior: Yes

Oestrous cyclicity (parental animals):

No data

Sperm parameters (parental animals):

No data

Litter observations:

STANDARDISATION OF LITTERS
- Performed on day 4 postpartum: [yes/no]: no data
PARAMETERS EXAMINED
The following parameters were examined in [F1 / F2 / F3] offspring: F2 generation
Litter size, litter weight, and sex ratio were examined.
GROSS EXAMINATION OF DEAD PUPS: yes

Statistics:

Food intake, litter size, litter weight, and body weight were assessed with the Bonferroni’ss multiple comparison test after the analysis of variance (ANOVA) or the Kruskal-Wallis test.
Sex ratio, survival, and behavioral developmental data were assessed with the Chi-square test (mxn) of frequency analysis. Movement activity data were assessed with the Mann-Whitney U-test of nonparametric method (Martin and Bateson, 1990). Multiple water T-maze performance data were assessed with the Sign-Wilcoxon test for trials and assessed with the Wilcoxon test within each treatment group.

Offspring viability indices:

Yes

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

Dam having underdeveloped mammary glands

Body weight and weight changes:

no effects observed

Description (incidence and severity):

The average body weight of male and female mice showed no significant adverse effects from the treatment during the preconception period, and the average body weight of dams showed no significant adverse effects during the gestation or lactation periods.

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

The average body weight of male and female mice showed no significant adverse effects from the treatment during the preconception period, and the average body weight of dams showed no significant adverse effects during the gestation or lactation periods.

Organ weight findings including organ / body weight ratios:

not specified

Histopathological findings: non-neoplastic:

not specified

Other effects:

no effects observed

Description (incidence and severity):

Test substance intake: There was no significant adverse effect of Sunset Yellow FCF on the average food intake during any period

Reproductive function: oestrous cycle:

not specified

Reproductive function: sperm measures:

not specified

Reproductive performance:

no effects observed

OTHER FINDINGS (PARENTAL ANIMALS):
No significant adverse effect on movement activity of exploratory behavior in either sex at eight weeks of age.

Dose descriptor:

NOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: no observed adverse effect

Clinical signs:

not specified

Mortality / viability:

mortality observed, treatment-related

Description (incidence and severity):

The survival indices at PND 0 showed viability indices at birth. The survivals were significantly reduced in the middle-dose group (600 mg/kg bw/day) of each sex.

Body weight and weight changes:

no effects observed

Description (incidence and severity):

No significant adverse effect was observed in litter weight

Sexual maturation:

no effects observed

Description (incidence and severity):

No significant adverse effect was observed in sex ratio at birth

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

not specified

Histopathological findings:

not specified

OTHER FINDINGS (OFFSPRING)
No significant adverse effect was observed in litter size

Dose descriptor:

LOAEL

Generation:

F2

Effect level:

600 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: Effects: Dam having underdeveloped mammary glands.The survival indices at PND 0 showed viability indices at birth. The survivals were significantly reduced in the middle-dose group (600 mg/kg bw/day) of each sex.

Dose descriptor:

NOAEL

Generation:

F2

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: see 'Remark'

Reproductive effects observed:

not specified

Conclusions:

The NOAEL for maternal toxicity study was considered to be 300 mg/kg bw/day whereas LOAEL was considered to be 600 mg/kg bw/day in Crj: CD-1 strain mouse when Sunset Yellow FCF was administered orally by diet.

Executive summary:

Sunset Yellow FCF was administered in the diet to 60 mice (10/sex/group) at dietary levels of 300, 600, and 1200 mg/kg bw/day the control groups (20 mice, 10/sex) were given basal diets (Nihon Clea, CE-2) to investigate for possible reproductive effect. Individual food intake of mice was measured during five divided periods - preconception (from five weeks of age to mating), mating (5 days), gestation (14 days), lactation (from birth to weaning), and F 1 generation (four to nine weeks of age).

During experiment adverse effects observed as dam having underdeveloped mammary glands. The survival indices at PND 0 showed viability indices at birth. The survivals were significantly reduced in the middle-dose group (600 mg/kg bw/day) of each sex.

And on other side the average body weight of male and female mice showed no significant adverse effects from the treatment during the preconception period, and the average body weight of dams showed no significant adverse effects during the gestation or lactation periods. No effects on food consumption, No significant adverse effect on movement activity of exploratory behavior in either sex at eight weeks of age.

Therefore, NOAEL for maternal toxicity study was considered to be 300 mg/kg bw/day whereas LOAEL was considered to be 600 mg/kg bw/day in Crj: CD-1 strain mouse when Sunset Yellow FCF was administered orally by diet.

Effect on fertility: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

300 mg/kg bw/day

Study duration:

chronic

Species:

rat

Quality of whole database:

The data is K2 level as the data has been obtained from the experimental study from the reliable journal 'Toxicology and industrial health'

Effect on fertility: via inhalation route

Endpoint conclusion:

no study available

Effect on fertility: via dermal route

Endpoint conclusion:

no study available

Additional information

WoE Summary of 2783-94-0 for toxicity to reproduction

Based on the various studies available with Klimish rating 2 and 4 for the target as well as the read across substances for CAS: 583-39-1 based on the category approach of organic functional group along with similar mechanistic approach and having structural similarities defined by QSAR toolbox.

The results for target as well as analogues are summarized as follows

Sr. No	End point	Value	Species	Route	Effects	Remarks
1.	LOAEL(parent)	600 mg/kg bw/day	Mouse (Crj: CD-1)	oral: feed	Dam having underdeveloped mammary glands.The survival indices at PND 0 showed viability indices at birth. The survivals were significantly reduced in the middle-dose group (600 mg/kg bw/day) of each sex.	Data from Publication of ‘Toxicology and industrial health’ for target CAS:2783-94-0
2.	NOAEL(F1 generation)	300 mg/kg bw/day	Mouse (Crj: CD-1)	oral: feed	The average body weight of male and female mice showed no significant adverse effects from the treatment during the preconception period, and the average body weight of dams showed no significant adverse effects during the gestation or lactation periods.No effects on food consumption,No significant adverse effect on movement activity of exploratory behavior in either sex at eight weeks of age.	Data from Publication of ‘Toxicology and industrial health’ for target CAS:2783-94-0
3.	NOAEL(F3 generation)	500 mg/kg bw/day	Rat (Charles River CD)	oral: feed	There were no compound related effects on fertility, gestation,on reproductive organs of females, or on organ weights among parents.There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats.	Data from study report of ‘International Research and Development Corporation’ for target CAS:2783-94-0
4.	NOEL	509 mg/kg bw/day	Rat (Crj: CD(SD))	oral: gavage	No any reprotoxic effects were observed.	Predicted data from QSAR for target CAS : 2783-94-0
5.	NOAEL(Parent)	100000 mg/kg diet	Rat (Sprague-Dawley)	oral: feed	not significantly affect gestation length,no malformations were seen,no significant changes in food consumption	Experimental data from publication ‘Toxicology’for read across CAS: 25956-17-6

 

 

Based on the studies summarized in the above table it can be observed that the endpoint lowest observed adverse effect level (LOAEL) found to be 600 mg/kg bw/day whereas no observed adverse effect level (NOAEL) value found to vary between 300 mg/kg bw/day to 100000 mg/kg diet based on the data from prediction as well as publication for target &read across substance. The effect observed on the above doses are-

  

* Dam having underdeveloped mammary glands. The survival indices at PND 0 showed viability indices at birth. The survivals were significantly reduced in the middle-dose group (600 mg/kg bw/day) of each sex.

* The average body weight of male and female mice showed no significant adverse effects from the treatment during the preconception period, and the average body weight of dams showed no significant adverse effects during the gestation or lactation periods. No effects on food consumption, No significant adverse effect on movement activity of exploratory behavior in either sex at eight weeks of age.

* There were no compound related effects on fertility, gestation, on reproductive organs of females, or on organ weights among parents. There were no compound related lesions in any tissue examined histologically, including kidneys and adrenal glands from parental rats.

* No any reprotoxic effects were observed.

* Not significantly affect gestation length,no malformations were seen,no significant changes in food consumption

 

Thus based on the above results it can be concluded that substance CAS: 2783-94-0 is expected to show the similar toxicological effect based on the effects observed on the other category members. Since low effective dose value (LOAEL) is 600 mg/kg bw/day based on this value it can be concluded that substance CAS: 2783-94-0 is considered to be not toxic to reproduction below the dose level of 600 mg/kg bw/day.

Short description of key information:

From the various data available and adopting the weight of evidence approach, it has been concluded that Sunset Yellow FCF is not likely to have reprotoxic effects.

Justification for selection of Effect on fertility via oral route:

The NOAEL for maternal toxicity study was considered to be 300 mg/kg bw/day whereas LOAEL was considered to be 600 mg/kg bw/day in Crj: CD-1 strain mouse when Sunset Yellow FCF was administered orally by diet.

Effects on developmental toxicity

Description of key information

From the various data available and adopting the weight of evidence approach, it has been concluded that Sunset Yellow FCF is not likely to have developmental toxic effects.

Link to relevant study records

Reference

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

4 (not assignable)

Principles of method if other than guideline:

The developmental effects of FD&C Yellow No. 6 on Pregnant Charles River CD rats and their offspring by oral (gavage) route at different dose levels, on days 6-15 of gestation, were examined.

GLP compliance:

not specified

Species:

rat

Strain:

other: Charles River CD

Route of administration:

oral: gavage

Type of inhalation exposure (if applicable):

not specified

Vehicle:

other: methocel and retinoic acid

Details on exposure:

FD&C Yellow No. 6 was administered by gavage at dose levels of 100, 300 or 1000 mg/kg bw/day to 140 female Charles River CD rats. Three negative control groups (20/group) received the vehicle control while one control group received the positive control (7.5% mg/kg bw/day retinoic acid). All females were dosed on days 6-15 of gestation.

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

9 days

Frequency of treatment:

Daily

Duration of test:

20 days

Remarks:

Doses / Concentrations:
100, 300 or 1000 mg/kg/day
Basis:
no data

No. of animals per sex per dose:

140 female Charles River CD rats.
Negative control:Three groups of 20 rats/group
Positive control: one group of 20 rats

Control animals:

yes

Details on study design:

Three negative control groups were maintained and administered 0.5% methocel, while one positive control group was maintained and administered 7.5% mg/kg bw/day of retinoic acid.

Details on maternal toxic effects:

Maternal toxic effects:no effects

Details on maternal toxic effects:
No effect was observed on any other maternal parameters like body weight, corpora lutea, empty implantation sites, early resorptions, late resorptions, and live or dead term fetuses.

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: no observed adverse effect

Details on embryotoxic / teratogenic effects:

Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No effect was observed on any fetal parameters evaluated like sex, external, internal and skeletal abnormalities. No teratogenicity was observed among the offspring.

Dose descriptor:

NOAEL

Effect level:

100 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Dose descriptor:

LOAEL

Effect level:

300 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: teratogenicity

Abnormalities:

not specified

Developmental effects observed:

not specified

Conclusions:

The teratogenicity of FD&C Yellow No. 6 was observed at dose concentrations of 0, 100, 300 or 1000 mg/kg bw/day, but no effect observed in parents and their offsprings at a dose concentration of 100 mg/kg bw/day. At a dose concentration of 300 mg/kg bw/day effects were observed as the mean weights of the offspring were decreased.
Therefore the ‘No observed adverse effect level (NOAEL) for teratogenicity was considered to be 100 mg/kg bw/day, and the “lowest observed adverse effect level (LOAEL)” was considered to be 300 mg/kg bw/day.

Executive summary:

The study was designed to investigate the teratogenic effects of FD&C Yellow No. 6 to pregnant rats by oral (gavage) route. FD&C Yellow No. 6 was administered by gavage at dose levels of 0,100, 300 or 1000 mg/kg bw/day to 140 female Charles River CD rats on days 6-15 of gestation.

 

No compound related effectswere observedon early or late resorptions, empty implantation sites, body weight or numbers of live or dead fetuses. No teratogenicity was observed among the offspring at concentration 100 mg/kg bw/day.

 

The mean weights of the offspring from the 300 and 1000 mg/kg bw/day groups were decreased when compared to the average fetus weight of the combined negative controls.

 

Therefore the ‘No observed adverse effect level (NOAEL) for teratogenicity was considered to be 100 mg/kg bw/day,andthe“lowest observed adverse effect level (LOAEL)” was considered to be 300 mg/kg bw/day when administered orally.

Effect on developmental toxicity: via oral route

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

100 mg/kg bw/day

Study duration:

subacute

Species:

rat

Quality of whole database:

The data is K4 level as the data has been obtained from the experimental study from the study report 'International Research and Development Corporation'.

Effect on developmental toxicity: via inhalation route

Endpoint conclusion:

no study available

Effect on developmental toxicity: via dermal route

Endpoint conclusion:

no study available

Additional information

WoE Summary of 2783-94-0 for developmental toxicity

Based on the various studies available with Klimish rating 2 and 4 for the target as well as the read across substances for CAS: 2783-94-0based on the category approach of organic functional group along with similar mechanistic approach and having structural similarities defined by QSAR toolbox.

The results for target as well as analogues are summarized as follows

Sr. No	End point	Value	Species	Route	Effects	Remarks
1.	NOAEL	100 mg/kg bw/day	Rat (Charles River CD)	oral: gavage	No compound related effects on early or late resorptions,empty implantation sites, body weight or numbers of live or dead fetuses. No teratogenicity was observed among the offspring.	Experimental data from study report ‘International Research and Development Corporation’for target CAS: 2783-94-0
2.	LOAEL	300 mg/kg bw/day	Rat (Charles River CD)	oral: gavage	The mean weights of the offspring from the 300 and 1000 mg/kg bw/day groups were decreased when compared to the average fetus weight of the combined negative controls.	Experimental data from study report ‘International Research and Development Corporation’for target CAS: 2783-94-0
3.	NOEL	94 mg/kg bw/day	Rat	oral: gavage	No any adverse effect was observed on fetus.	Predicted data from QSAR for target CAS: 2783-94-0
4.	NOAEL	1000mg/kg bw/day	Rat (Osborne-Mendel)	oral: gavage	Skeletal anomaly was not reported at the higher dose levels. No other soft-tissue or sternebral variations were reported.	Experimental data from study report ‘Food and Chemical Toxicology’for read across CAS: 25956-17-6
5.	NOAEL	1000mg/kg bw/day	Rat (HanIbm: WIST)	oral: gavage	No abnormalities were observed in any foetuses. In the foetuses of group treated at 1000 mg/kg bw/day	Experimental data from study report ‘European Commission opinion report’for read across CAS:633-69-5

  

Based on the studies summarized in the above table it can be observed that the endpoint lowest observed adverse effect level (LOAEL) found to be 300 mg/kg bw/day whereas no observed adverse effect level (NOAEL) value found to vary between 94 mg/kg bw/day to 1000mg/kg bw/day based on the data from prediction as well as publication for target &read across substance. The effect observed on the above doses are-

  

* The mean weights of the offspring from the 300 and 1000 mg/kg bw/day groups were decreased when compared to the average fetus weight of the combined negative controls.

* No compound related effects on early or late resorptions,empty implantation sites, body weight or numbers of live or dead fetuses. No teratogenicity was observed among the offspring.

* No any adverse effect was observed on fetus.

* Skeletal anomaly was not reported at the higher dose levels. No other soft-tissue or sternebral variations were reported.

* No abnormalities were observed in any foetuses. In the foetuses of group treated at 1000 mg/kg bw/day. 

Thus based on the above results it can be concluded that substance CAS: 2783-94-0 is expected to show the similar toxicological effect based on the effects observed on the other category members. Since low effective dose value (LOAEL) is 300 mg/kg bw/day based on this value it can be concluded that substance CAS: 2783-94-0 is considered to be not toxic to developmental toxicity below the dose level of 300 mg/kg bw/day.

Justification for selection of Effect on developmental toxicity: via oral route:

The teratogenicity of FD&C Yellow No. 6 was observed at dose concentrations of 0, 100, 300 or 1000 mg/kg bw/day, no effect observed in parents and their offsprings at dose concentration 100 mg/kg bw/day and up to dose concentration 300 mg/kg bw/day effects observed as the mean weights of the offspring was decreased.

Therefore the ‘No observed adverse effect level (NOAEL) for teratogenicity was considered to be 100 mg/kg bw/day and “lowest observed adverse effect level (LOAEL)” was considered to be 300 mg/kg bw/day.                                                                            

Justification for classification or non-classification

From the NOAEL and LOAEL values obtained for Sunset Yellow FCF for toxicity to reproduction as well as developmental toxicity, it is considered that the chemical does not qualify for classification as a reproductive or developmental toxicant.