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Administrative data

Key value for chemical safety assessment

Effects on fertility

Link to relevant study records
Reference
Endpoint:
one-generation reproductive toxicity
Remarks:
based on generations indicated in Effect levels (migrated information)
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
Reference:
Composition 0
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 421 (Reproduction / Developmental Toxicity Screening Test)
Principles of method if other than guideline:
Teratogenicity study of Fluorescein sodium in rabbits
GLP compliance:
not specified
Limit test:
no
Test material information:
Composition 1
Species:
rabbit
Strain:
New Zealand White
Sex:
female
Details on test animals and environmental conditions:
EST ANIMALS
- Source: No data available
- Age at study initiation: (P) x wks; (F1) x wks No data available
- Weight at study initiation: (P) Males: x-x g; Females: x-x g; (F1) Males: x-x g; Females: x-x g: 4.5 kg
- Fasting period before study: No data available
- Housing: No data available
- Diet (e.g. ad libitum): No data available
- Water (e.g. ad libitum): No data available
- Acclimation period: No data available

ENVIRONMENTAL CONDITIONS
- Temperature (°C): No data available
- Humidity (%):No data available
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): No data available
Route of administration:
intravenous
Type of inhalation exposure (if applicable):
not specified
Vehicle:
other: 10% water solution (Funduscein)
Details on mating procedure:
- M/F ratio per cage: No data available
- Length of cohabitation: No data available
- 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): No data available
- Any other deviations from standard protocol: Nineteen rabbits immediately after copulation with male rabbits of the same species were used.
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
3 days (5,6 and 8 day after copulation)
Frequency of treatment:
Once daily on 5,6 and 8 day after copulation
Details on study schedule:
No data available
Remarks:
Doses / Concentrations:
1.4 ml (2241.4 mg/kg)
Basis:
actual ingested
No. of animals per sex per dose:
Total: 19
0 ml: 4 female
2241.4 mg/kg: 15 female
Control animals:
yes, concurrent vehicle
Details on study design:
No data available
Positive control:
No data available
Parental animals: Observations and examinations:
Survival and clinical sign were examined.
Estrous cyclicity (parental animals):
Any irriggularty in Estrous cyclicity were examined.
Sperm parameters (parental animals):
No data available
Litter observations:
Delayed appearance of birth defects were examined.
Postmortem examinations (parental animals):
No data available
Postmortem examinations (offspring):
Gross pathology and histopathology were examined.
Statistics:
The difference between the number of offspring that died at one month in the experimental vs the control group was analyzed with the Chi-square test. The probability that the distribution occurred by chance was found to be at the 5% level of significance (P = .05).
Reproductive indices:
No data available
Offspring viability indices:
Viability till four weeks were observed
Clinical signs:
no effects observed
Body weight and weight changes:
not specified
Food consumption and compound intake (if feeding study):
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Other effects:
not specified
Reproductive function: estrous cycle:
not specified
Reproductive function: sperm measures:
not specified
Reproductive performance:
no effects observed
No effect on survival were observed in trated rats as compared to control.

Clinical signs:
No abortion or miscarriage were observed in trated rats as compared to control.

Body weight and food consumption: No data available

Test substance intake: No data available

Reproductive function: estrous cycle: No data available

Reproductive function: sperm measures: No data available

Reproductive performance:No effect on live-born offspring and stillbirth were observed in treated rats as compared to control.

Organ weights No data available

Gross pathology: No data available

Histopathology: No data available

other findings No data available
Dose descriptor:
NOAEL
Effect level:
2 241.4 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: No effect on survival, clinical sign and Reproductive performance
Clinical signs:
no effects observed
Mortality / viability:
no mortality observed
Body weight and weight changes:
not specified
Sexual maturation:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
no effects observed
Histopathological findings:
no effects observed
Mortality:
No effect on live-born offspring were observed in trated rats as compared to control.

At 4 week, 14 offspring (19%) from four mothers subsequently died.
The observed effect were due to poor mothers not due to treatment.

Clinical signs:
One stillbirth and all other offspring were normal and no delayed appearance of birth defects were observed in offspring as compared to control.

Body weight and food consumption: No data available

Test substance intake: No data available

Reproductive function: estrous cycle: No data available

Reproductive function: sperm measures: No data available

Reproductive performance: No data available

Organ weights: No data available

Gross pathology:No gross pathological changes were observed in offspring of treated rats.

Histopathology:No Soft tissue and Visceral abmormalities were observed in offspring of treated rats.

Dose descriptor:
NOAEL
Generation:
F1
Effect level:
2 241.4 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on survival, clinical sign, Gross pathology and histopathology
Reproductive effects observed:
not specified
Conclusions:
NOAEL was considered to be 2241.4 mg/kg when New Zealand female Rabbits were treated with Fluorescein sodium.
Executive summary:

In a Teratogenicity study,New Zealand female Rabbits were treated with Fluorescein sodium in the concentration of 2241.4 mg/kg intravenouslly in 10% water solution (Funduscein) on day 5,6 and 8 day after copulation. No effect was observed on survival and no abortion or miscarriages were observed in treated rats as compared to control. No effect on live-born offspring and stillbirth were observed in treated rabbits as compared to control.  No effect on live-born offspring was observed. At 4 week, 14 offspring (19%) from four mothers subsequently died. The observed effects were due to poor mothers and not due to treatment. One stillbirth and all other offspring were normal and no delayed appearances of birth defects were observed in offspring as compared to control. In addition, no gross pathological and histopathological changes were observed in offspring of treated rabbits as compared to control. Therefore, NOAEL was considered to be 2241.4 mg/kg for F0 and F1 generation when New Zealand female Rabbits were treated with Fluorescein sodium intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 day and day 13, 15 and 16 day after copulation.

Effect on fertility: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
2 241.4 mg/kg bw/day
Study duration:
subacute
Species:
rabbit
Quality of whole database:
Data is Klimish 2 and from peer reviewed journal
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: 

Data available for target Sodium fluorescein (CAS no 5185-47-8) for reproductive toxicity are summarized as below.

In a study conducted by Mcenerneyet al(1977), Teratogenicity was evaluated in New Zealand female Rabbits treated with Fluorescein sodium in the concentration of 2241.4 mg/kg intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 and day 13, 15 and 16 after copulation. No effect was observed on survival and no abortion or miscarriages were observed in treated rats as compared to control. No effect on live-born offspring and stillbirth were observed in treated rabbits as compared to control.  No effect on live-born offspring was observed. At 4 week, 14 offspring (19%) from four mothers subsequently died. The observed effects were due to poor mothers and not due to treatment. One stillbirth and all other offspring were normal and no delayed appearances of birth defects were observed in offspring as compared to control. In addition, no gross pathological and histopathological changes were observed in offspring of treated rabbits as compared to control. Therefore, NOAEL was considered to be 2241.4 mg/kg for F0 and F1 generation when New Zealand female Rabbits were treated with Fluorescein sodium intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 day and day 13, 15 and 16 day after copulation.

In a study conducted by Burnetiet al(1986), Teratogenic toxicity was evaluated in Dutch Belted female Rabbits treated with D&C Yellow no 8 in the concentration of 0, 30, 100 and 250 mg/kg body weight/day by oral gavage. One rabbits died each in 30 and 250 mg/kg bw/day including control. One rabbit aborted on day 28 of gestation at 250 mg/kg bw/day as compared to control. Pneumonia was observed in three rabbit which were died. Orange discoloration of the urine was observed in all the treated rabbits between days 7 and 28 of gestation as compared to control. No effect on body weight and body weight gain and No effect on Post-implantation loss/dam, Total implantations/dam and Corpora lutea/dam were observed in treated rabbits as compared to control. Similarly, There were no biologically meaningful or statistically significant differences in the number of litters, Foetal sex and gross pathological changes were observed in treated rabbits as compared to control. In addition, No effect on viability of foetus, foetal body weight and sex of foetus were observed as compared to control. At 30 and 100 mg/kg be/day, malformations were observed in two foetuses from two litters. However, these values fell within the ranges of historical control data. There were no biologically meaningful or statistically significant differences in number of fetuses with malformations, number of foetuses or litters with developmental variations were observed in foetuses of any of the treated groups. Therefore, NOAEL was considered to be 250 mg/kg body weight /day for F0 and F1 generation when Dutch Belted female rabbits were treated with D&C Yellow no 8 orally by gavage from day 6 to 27 of gestation.

In a above similar reference, Teratogenic toxicity was evaluated in CD Sprague-Dawley female rats treated with D&C Yellow no. 8 in the concentration of 0, 100, 500 and 1500 mg/kg body weight/day by oral gavage. Six rats died during the dosing period and at 1500 mg/kg bw/day and Orange discoloration of the urine was observed in all the treated rats as compared to control. Similarly, green discoloration of the amniotic fluid and green colored small intestines were observed at 1500 mg/kg bw/day and green discoloration of the amniotic fluid was observed at 100 and 500 mg/kg bw/day treated rats as compared to control. No effect on Post-implantation loss/dam, Total implantations/dam, Corpora lutea/dam and There were no biologically meaningful or statistically significant differences in the number of litters and Foetal sex of treated rats were observed as compared to control. In addition, No effect on viability of foetuse, number of fetuses with malformations, number of foetuses or litters with developmental variations were observed in any of the treated groups. Slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14thrib(s) was observed in feotus of 1500 mg/kg bw/day treated rats as compared to control, but the observed effect fell within the ranges of historical control data. Therefore, NOAEL was considered to be 1500 mg/kg body weight /day for F0 and F1 generation when CD Sprague-Dawley female rats were treated with D&C Yellow no. 8 orally by gavage for 14 days.

Thus, based on the above available data for target sodium fluorescein (CAS no 518-47-8) from peer reviewed journals is not likely to be classified as a reproductive toxicant in rabbits and rats.


Short description of key information:
NOAEL was considered to be 2241.4 mg/kg for F0 and F1 generation when New Zealand female Rabbits were treated with Fluorescein sodium

Justification for selection of Effect on fertility via oral route:
NOAEL was considered to be 2241.4 mg/kg for F0 and F1 generation when New Zealand female Rabbits were treated with Fluorescein sodium intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 day and day 13, 15 and 16 day after copulation.

Effects on developmental toxicity

Description of key information
NOAEL was considered to be 1500 mg/kg bw F0 and F1 generation when Sprague-Dawley Female rats were treated with D & C Yellow No. 8 
Link to relevant study records
Reference
Endpoint:
developmental toxicity
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- reivewed journal
Reference:
Composition 0
Principles of method if other than guideline:
The teratogenic potential was determined in Sprague dawley rats after the administration of D & C Yellow No. 8 by gavage
GLP compliance:
not specified
Limit test:
no
Test material information:
Composition 1
Species:
rat
Strain:
Sprague-Dawley
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Portage, Michigan
- Age at study initiation: 14 weeks
- Weight at study initiation:No data available
- Fasting period before study:No data available
- Housing:No data available
- Diet (e.g. ad libitum):No data available
- Water (e.g. ad libitum):No data available
- Acclimation period: 4 weeks

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 72 ± 4F
- Humidity (%): 50 ± 15%
- Air changes (per hr): No data avaialble
- Photoperiod (hrs dark / hrs light): 2-hrlight/dark cycle

IN-LIFE DATES: From: To:No data available
Route of administration:
oral: gavage
Type of inhalation exposure (if applicable):
not specified
Vehicle:
not specified
Details on exposure:
From day 6-19 days of gestation
Analytical verification of doses or concentrations:
not specified
Details on mating procedure:
- M/F ratio per cage: No data available
- Length of cohabitation: No data available
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy Copulatory plug or presence of sperm in vaginal washings.
- 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
- After successful mating each pregnant female was caged (how): They were housed individually and offered Purina Certified Rodent Chow No. 5002 and water ad lib.
- Any other deviations from standard protocol: Not mentioned
Duration of treatment / exposure:
14 days
Frequency of treatment:
Daily
Duration of test:
days 6-19 of gestation
No. of animals per sex per dose:
Total: 100
0 mg/Kg bw: 25
100 mg/Kg bw: 25
500 mg/Kg bw: 25
1500 mg/Kg bw: 25
Control animals:
yes, concurrent vehicle
Maternal examinations:
Caesarean sections were performed on all surviving females killed with carbon dioxide on gestation day 20 The number and location of viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea were recorded.
Ovaries and uterine content:
Gravid and Nongravid uteri were examined.
Fetal examinations:
Foetal sex ratio, foetal body weight, gross pathology and histopathology were examined.
Statistics:
Differences in the Ibctal scx distribution and the number of litters with malformations between control and treated groups wcre compared using the Chi-square test criterion with Yates" correction for 2 x 2 contingency lables and, or Fisher's exact probability test as described by Sicgel (1956). The numbers of early and late resorptions, dead foetuses and post-implantation losses were compared between groups by the Mann Whitney U test as described by Siegel (1956) and Well (1970). The mean numbers of viable foetuses, total implantations, corpora lutca and mean t\)etal weights were compared between groups by analysis of variance (oneway classification), Bartlett's test l\~r homogeneity of variances, and the appropriate t test (for equal or unequal variances) as described by Steel & Torric (1960) using Dunnctt's multiple comparison tables (1964).
Indices:
Dams with viable foetuses, Viable foetuses/dam, Post-implantation loss, Total implantations and Corpora lutea were examined.
Historical control data:
No data available
Details on maternal toxic effects:
Maternal toxic effects:yes

Details on maternal toxic effects:
Mortality:
when treated wtih 1500 mg/kg bw, Six rats died as compared to control.

Clinical sign:
Orange discoloration of the urine was observed in 100, 500 and 1500 mg/kg bw treated rats.

Body weigth gain:
Slight decrease in body weight gain were observed in 1500 mg/kg bw treated rats as compared to control.

Reproductive indices:
No effect on dams with viable foetuses, viable foetuses/dam, post-implantation loss, total implantations, corpora lutea and number of litters with malformations were examined.

Gross pathology:
Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats in the 100, 500 and 1500 mg/kg/day treated rats as compared to control.
Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day
Based on:
test mat.
Basis for effect level:
other: maternal toxicity
Details on embryotoxic / teratogenic effects:
Embryotoxic / teratogenic effects:no effects

Details on embryotoxic / teratogenic effects:
No significant effect on Foetal sex ratio, foetal body weight, gross pathology and histopathology were observed.
Dose descriptor:
NOAEL
Effect level:
1 500 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on foetal body weight, sex retio, gross pathology and histopathology
Abnormalities:
not specified
Developmental effects observed:
not specified

Mean maternal body weights changesfor rats given D & (" Yellow No. 8 by gavage on days 6 -19 of gestation

 

Day of gestation

       No. of foetuses (no. of litters)

 

0

100mg/kg

500mg/kg

1500mg/kg

0

246±145    

249 ± I3.7

5249±14.7

241±12. 4

6

272±16.8

271 ± 14.4

276±14.5

263 ± 12.2

9

279±18,1

278 ± 15.9

283±16.7

269 ±16.5

12

294±20.1

292 ± 17.5

296 ± 16.1

282 ±16.9

16

321±23.4

320 ± 18.9

323 ± 20.4

300 ± 30.7

20

379±30.0

378 ±21.2

374 ±32.2

359 ±33.3

Conclusions:
NOAEL was considered to be 1500 mg/kg bw when Sprague-Dawley Female rats were treated with D & C Yellow No. 8 orally by gavage from day 6-19 of gestation.
Executive summary:

In a teratogenic potential study, Sprague-Dawley Female rats were treatedw with D & C Yellow No. 8 in the concentration of 0, 100, 500 and 1500mg/kg body weight orally by gavage. Six rats died at 1500 mg/kg bw as compared to control. Orange discoloration of the urine was observed in 100, 500 and 1500 mg/kg bw treated rats and Slight decrease in body weight gain were observed in 1500 mg/kg bw treated rats as compared to control. Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats in the 100, 500 and 1500 mg/kg/day treated rats as compared to control. But, no effect on dams with viable foetuses, viable foetuses/dam, post-implantation loss, total implantations, corpora lutea and number of litters with malformations were examined in treated rats as compared to control. In addition, No significant effect on Foetal sex ratio, foetal body weight, gross pathology and histopathology were observed. Therefore, NOAEL was considered to be 1500 mg/kg bw when Sprague-Dawley Female rats were treated with D & C Yellow No. 8 orally by gavage from day 6-19 of gestation.

Based on the observations these doses did not result in evidence of maternal toxicity or adverse effects on foetal development.

Effect on developmental toxicity: via oral route
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 500 mg/kg bw/day
Study duration:
subacute
Species:
rat
Quality of whole database:
Data is Klimish 2 and from peer reviewed journal
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

Developmental toxicity:

Data available for target Sodium fluorescein (CAS no 5185-47-8) for developmental toxicity are summarized as below.

In a study conducted by Burnetiet al(1986), teratogenic potential was evaluated in Sprague-Dawley Female rats by using D & C Yellow No. 8 in the concentration of 0, 100, 500 and 1500mg/kg body weight orally by gavage. Six rats died at 1500 mg/kg bw as compared to control. Orange discoloration of the urine was observed in 100, 500 and 1500 mg/kg bw treated rats and Slight decrease in body weight gain were observed in 1500 mg/kg bw treated rats as compared to control. Green discoloration of the amniotic fluid was observed in 1, 10 and 16 rats in the 100, 500 and 1500 mg/kg/day treated rats as compared to control. But, no effect on dams with viable foetuses, viable foetuses/dam, post-implantation loss, total implantations, corpora lutea and number of litters with malformations were examined in treated rats as compared to control. In addition, No significant effect on Foetal sex ratio, foetal body weight, gross pathology and histopathology were observed. Therefore, NOAEL was considered to be 1500 mg/kg bw F0 and F1 generation when Sprague-Dawley Female rats were treated with D & C Yellow No. 8 orally by gavage from day 6-19 of gestation.

In a above similar study, teratogenic potential was evaluated in Dutch Belted Female rabbits by using D & C Yellow No. 8 in the concentration of 0, 30, 100 and 250 mg/kg body weight orally by gavage. One rabbit died in 30, 25 and in control due to pneumonia and one rabbit aborted on day 28 of gestation in 250 mg/kg bw. Green discoloration of the urine was observed in 30, 100 and 250 mg/kg bw treated rabbits. No effect on body-weight gains were observed in treated rabbits. Similarly, No statistically significant effect on numbers of corpora lutea, total implantations and early or late resorptions were observed in treated rabbits as compared to control. In addition, No statistically significant effect on viable foetuses, foetal sex, distribution or mean foetal body weight, external, visceral and skeletal foetal malformations and developmental variations were observed in treated rabbits. Only malformations observed occurred in two foetuses from two litters in both the 30 and 100 mg/kg/day groups and these have been observed in historical control data. Therefore, NOAEL was considered to be 250 mg/kg bw for F0 and F1 generation when Dutch Belted Female rabbits were treated with D & C Yellow No. 8 orally by gavage from day 6-27 of gestation.

 

In a study conducted by Salemet al(1979), Teratogenicity was evaluated in CD female rats by using sodium fluorescein in the concentration of 0 and 10000 mg/kg (10 %) by intravenous injections under light ether anesthesia into the saphenous vein on days 1, 6, 12 and 18 of pregnancy (4 different female rats per day). No statistically significant effect on number of pups per litter was observed as compared to control. In addition, No statistically significant effect on overt abnormalities, average length (cm) or weight (g) of the pups and gross external and internal organs of pups of treated rats as compared to control. Therefore, NOAEL was considered to be 10000 mg/kg for F0 and F1 generation when CD female rats were treated with sodium fluorescein intravenously into the saphenous vein on days 1, 6, 12 and 18 of pregnancy (4 different female rats per day).

In a study conducted by Mcenerneyet al(1977), Teratogenicity was evaluated in New Zealand female Rabbits treated with Fluorescein sodium in the concentration of 2241.4 mg/kg intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 and day 13, 15 and 16 after copulation. No effect was observed on survival and no abortion or miscarriages were observed in treated rats as compared to control. No effect on live-born offspring and stillbirth were observed in treated rabbits as compared to control.  No effect on live-born offspring was observed. At 4 week, 14 offspring (19%) from four mothers subsequently died. The observed effects were due to poor mothers and not due to treatment. One stillbirth and all other offspring were normal and no delayed appearances of birth defects were observed in offspring as compared to control. In addition, no gross pathological and histopathological changes were observed in offspring of treated rabbits as compared to control. Therefore, NOAEL was considered to be 2241.4 mg/kg for F0 and F1 generation when New Zealand female Rabbits were treated with Fluorescein sodium intravenouslly in 10% water solution (Funduscein) on day 5, 6 and 8 day and day 13, 15 and 16 day after copulation.

Thus, based on above available data for target sodium fluorescein (CAS no 518-47-8) from peer reviewed journals is not likely to be classified as a developmental toxicant in rabbits and rats.


Justification for selection of Effect on developmental toxicity: via oral route:
NOAEL was considered to be 1500 mg/kg bw F0 and F1 generation when Sprague-Dawley Female rats were treated with D & C Yellow No. 8 orally by gavage from day 6-19 of gestation.

Justification for classification or non-classification

sodium fluorescein (CAS no 518-47-8) from peer reviewed journals is not likely to be classified as a reproductive and developmenal toxicant in rabbits and rats.