Disodium 2-(2,4,5,7-tetrabromo-6-oxido-3-oxoxanthen-9-yl)benzoate

Developmental toxicity / teratogenicity

Administrative data

Endpoint:

developmental toxicity

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Justification for type of information:

Data from peer reviewed journal

Data source

Reference

Reference Type:

publication

Title:

Developmental Toxicity and Psychotoxicity Evaluation of test chemical in Rats

Author:

Charles V. Vorhees et.al

Year:

2018

Bibliographic source:

Arch Toxicol (1983)

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

other: As mentioned below

Principles of method if other than guideline:

Test chemicalwas given in diet to Sprague-Dawley rats From parent to offspring generation to evaluate the developmental toxicity and psychotoxicity effects.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

EC Number:

240-474-8

EC Name:

Disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

Cas Number:

16423-68-0

Molecular formula:

C20H8I4O5.2Na

IUPAC Name:

disodium 2-(2,4,5,7-tetraiodo-6-oxido-3-oxoxanthen-9-yl)benzoate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material (as cited in study report): Erythrosine
- Molecular formula (if other than submission substance): C20-H6-I4-Na2-O5
- Molecular weight (if other than submission substance): 879.86
- InChl Key (if other than submission substance): RAGZEDHHTPQLAI-UHFFFAOYSA-L
-Substance type- Organic
- Physical state: Solid (powder)
- Analytical purity:91%
- Impurities (identity and concentrations):9 % impurities

Test animals

Species:

rat

Strain:

Sprague-Dawley

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source:Laboratory Supply Co., Indianapolis, Indiana, USA
- Age at study initiation:90-110 days of age of the offspring
- Weight at study initiation:Male (200-220 g) and female (200-220 g)
- Fasting period before study:No data available
- Housing:No data available
- Diet (e.g. ad libitum):ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period:5 days

Administration / exposure

Route of administration:

oral: feed

Type of inhalation exposure (if applicable):

not specified

Vehicle:

not specified

Details on exposure:

Details on exposure
PREPARATION OF DOSING SOLUTIONS: Test chemical mixed in powdered Purina rat chow.

VEHICLE
- Justification for use and choice of vehicle (if other than water):Purina rat chow were used.
- Concentration in vehicle: 0.0, 0.25, 0.5 and 1.0 %

Analytical verification of doses or concentrations:

not specified

Details on mating procedure:

- Impregnation procedure: [artificial insemination / purchased timed pregnant / cohoused]:cohoused
- If cohoused:
- M/F ratio per cage:1:1
- Length of cohabitation:Till the confirmation of breeding
- Further matings after two unsuccessful attempts: [no / yes (explain)]:No data
- Verification of same strain and source of both sexes: [yes / no (explain)]:No data
- Proof of pregnancy: [vaginal plug / sperm in vaginal smear] referred to as [day 0 / day 1] of pregnancy:The presence of sperm in the daily vaginal lavage of the females and designated as day zero of gestation.
- Any other deviations from standard protocol:No data

Duration of treatment / exposure:

For male : fed diets containing the dye for 2 weeks.
For female: The dye was fed to each treatment group for 2 weeks prior to breeding, 1-14 days during breeding , then to the females during gestation and lactation.

Frequency of treatment:

Daily

Duration of test:

From parent to offspring generation

Doses / concentrations

Remarks:

0.0, 0.25, 0.5, or 1.0%( 250, 500 and 1000 mg /kg bw/day )

No. of animals per sex per dose:

Total: 196
0.0%: 19 male, 19 female
0.25%: 22 male, 22 female
0.5 %: 18 male, 18 female
1.0%: 21 male, 21 female
50 mg/kg (positive control): 18 male, 18 female

Control animals:

yes

Details on study design:

No data available

Examinations

Maternal examinations:

BODY WEIGHT: Yes
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
OTHER:females for reproductive success.

Ovaries and uterine content:

No data available

Fetal examinations:

Cincinnati Psychoteratogenicity Screening Test Battery, plus weight, food consumption, physical landmarks of development, and brain weight.

Statistics:

Analysis of variance for unequal group sizes, with split-plot designs where repeated measures were made, were performed on the majority of the data. Duncan a posteriori multiple range comparison tests for unequal group sizes (Kramer 1956) were used on those measures on which significant F-ratios occurred. Frequency data (e.g., mortality) were analyzed by Fisher's test for uncorrelated proportions (Guilford 1965). On preweaning tests, data on individual subjects were averaged for the entire litter or within sex and the litter was, therefore, the unit of analysis. On postweaning tests, only one male and one female was used from each litter on any given test,therefore, litter was again the unit of analysis, but no litter averaging was required. Because of the large number of tests done, and therefore, the number of statistical tests required, effects were only accepted as significant if the overall test (i.e., usually an F-test) occurred at a level ofp < 0.01 or beyond. Follow-up a posteriori individual group comparisons, however, were less stringent (p < 0.05) under the assumption that the F-test provided an adequate protection level when used at p < 0.01.

Indices:

No data available

Historical control data:

No data available

Results and discussion

Results: maternal animals

General toxicity (maternal animals)

Clinical signs:

not specified

Dermal irritation (if dermal study):

not specified

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

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):

no effects observed

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Maternal developmental toxicity

Number of abortions:

no effects observed

Pre- and post-implantation loss:

no effects observed

Total litter losses by resorption:

not specified

Early or late resorptions:

not specified

Dead fetuses:

no effects observed

Changes in pregnancy duration:

no effects observed

Changes in number of pregnant:

no effects observed

Other effects:

not specified

Effect levels (maternal animals)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day

Based on:

test mat.

Basis for effect level:

other: maternal toxicity

Remarks on result:

other: No reproductive toxic effects were observed

Maternal abnormalities

Abnormalities:

not specified

Localisation:

not specified

Results (fetuses)

Fetal body weight changes:

no effects observed

Reduction in number of live offspring:

no effects observed

Changes in sex ratio:

no effects observed

Changes in litter size and weights:

no effects observed

Changes in postnatal survival:

no effects observed

External malformations:

no effects observed

Skeletal malformations:

not specified

Visceral malformations:

not specified

Other effects:

no effects observed

Description (incidence and severity):

No advers effect were observed onPsychotoxicity of treated pups as compared to control.

Effect levels (fetuses)

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

fetal/pup body weight changes

changes in postnatal survival

external malformations

Remarks on result:

other: No developmental toxic effects were observed

Fetal abnormalities

Abnormalities:

not specified

Overall developmental toxicity

Developmental effects observed:

not specified

Treatment related:

not specified

Any other information on results incl. tables

Offspring mortality expressed as a % of those alive at the beginning of each phase

	Treatment	Days of age			No. Of liveoffspring after culling on day 1
		1-21	22-24	25-90
Experiment 1	R3 1.0	9.3^a	0.0	0.7	161
	R3 0.5	10.2^a	0.0	1.5	147
	R3 0.25	1.5	0.0	0.0	128
	R3 0.0	3.2	0.0	0.8	412
	R3 HU	8.6^a	0.0	1.5	219
Experiment 2	R3 1.0	9.9^a	0.0	6.9	161
	R3 0.5	15.9^a	0.0	3.4	157
	R3 0.25	23.3	0.0	1.0	193
	R3 0.0	22.5	0.0	0.0	129
^aSignificantly different from the 0% dye exposure group, p < 0.01

Swimming development ratings (mean _+ SE)

	Treatment	n^a	Swimming-direction day 6	n^a	Swimming-angle day 10
Experiment 1	R3 1.0	14	1.9±0.1	12	2.4±0.2*
	R3 0.5	13	1.6±0.1	13	2.1±0.1
	R3 0.25	12	1.8±0.1	11	2.4±0.3*
	R3 0.0	35	1.7±0.1	35	1.8±0.1
	R3 HU	19	1.7±0.1	19	2.1±0.2
Experiment 2	R3 1.0	14	2.2±0.2**	14	1.9±0.1
	R3 0.5	13	2.6±0.2	13	2.1±0.2*
	R3 0.25	14	2.6±0.1	14	2.3±0.2**
	R3 0.0	11	2.6±0.2	11	1.9±0.2
* F-test p < 0.01, individual group comparison to R3 0.0 group, p < 0.05 ** F-test p < 0.01, individual group comparison to R3 0.0 group, p < 0.01 an represents the number of litters tested, n varies slightly due to missing data points n = 35 in the R3 0.0 group because of controls from a previous similar experiment

Applicant's summary and conclusion

Conclusions:

NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawley male and female rat were exposed to test chemical .

Executive summary:

The reproductive and developmental effects of test chemical have been investigated in a 1-generation study in rats. The test chemical was administered at dietary dose levels of 0, 0.25, 0.5and 1.0 % (250, 500 and 1000 mg /kg bw/d) togroups of male and female Sprague-Dawley rats for two weeks prior to breeding, 1-14 days during breeding, then to females during gestation and lactation. The experimental diets were freely available to the offspring throughout postnatal development up to the age of 90-110 days. Body weights were measured weekly except during breeding. On the dayfollowing birth, all litters were examined and data collected on litter size, sex distribution,weight, and number of dead or malformed offspring. A variety of behavioural tests were determined in the offspring. Further, brainweights of day 90 were determined in the offspring. The experiment was replicated after2 years using the same exposure regimen.

The test chemical produced no effects on paternal or offspring weight or food consumption. No significant effects of any of the measures ofreproductive performance (proportion of females producing litters to those bred, thenumber of small litters, gestation length, litter size, sex ratio) were observed. No malformations were seen upon external examination. Preweaning offspring mortality was significantly increased at the 1.0 % and 0.5 % dose levels in the first experiment, but not inthe second. No adverse effects on offspring growth or adult regional brain weight were observed.No adverse effects were found in the present experiments on parental weight, food consumption, or reproductive performance.Therefore,NOAEL was considered to be 1.0 % (1000 mg/kg/day) for F0 and F1 generation when Sprague-dawleymale andfemale rat were exposed to test chemical .