Disodium 2-(3-oxo-6-oxidoxanthen-9-yl)benzoate

Endpoint summary

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)

Principles of method if other than guideline:

Teratogenic toxicity study of D&C YELLOW NO. 8 in rats

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Breeding Laboratories
- Age at study initiation: 18 weeks
- Weight at study initiation: 246 to 379 g
- Fasting period before study: No data available
- Housing: Animals were housed individually and identified by ear tag.
- Diet (e.g. ad libitum): Purina Certified Rodent Chow No. 5002, ad libitum
- Water (e.g. ad libitum): water, ad lib.
- Acclimation period: 4 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22.22 ± 15
- Humidity (%):50 ± 15%
- Air changes (per hr): No data available
- Photoperiod (hrs dark / hrs light): 12-hr light/dark cycle.

IN-LIFE DATES: From: To: No data available

Route of administration:

oral: gavage

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS: Doses were prepared daily in an aqueous solution at 100, 500 or 1500 mg/kg body weight.

DIET PREPARATION
- Rate of preparation of diet (frequency): No data available
- Mixing appropriate amounts with (Type of food): No data available
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): Water
- Concentration in vehicle: 0, 100, 500 or 1500 mg/kg body weight.
- Amount of vehicle (if gavage): 10 ml/kg
- Lot/batch no. (if required): No data available
- Purity: No data available

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

14 days

Frequency of treatment:

Daily

Remarks:

Doses / Concentrations:
0, 100, 500 or 1500 mg/kg body weight.
Basis:
nominal in water

No. of animals per sex per dose:

Total: 100
0 mg/kgbw/day:25 female
100 mg/kgbw/day:25 female
500 mg/kgbw/day:25 female
1500 mg/kgbw/day:25 female

Control animals:

yes, concurrent vehicle

Details on study design:

- Dose selection rationale: No data available
- Rationale for animal assignment (if not random): Mated females were assigned to the control group or to one of the three treatment groups of 25 animals each.
- Rationale for selecting satellite groups: No data available
- Post-exposure recovery period in satellite groups: No data available
- Section schedule rationale (if not random): 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: Survival was observed.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Daily

BODY WEIGHT: Yes
- Time schedule for examinations: On days 6, 9, 12 and 16 of gestation

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 available

WATER 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: No data available
- Time schedule for collection of blood: No data available
- Anaesthetic used for blood collection: No data available
- Animals fasted: No data available
- How many animals: No data available
- Parameters checked in table [No.?] were examined. No data available

CLINICAL CHEMISTRY: No data available
- Time schedule for collection of blood: No data available
- 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 available

OTHER: No data available

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
The thoracic and abdominal cavities and the internal organs of the dams were examined for gross morphological changes.

HISTOPATHOLOGY: No data available

Other examinations:

The number and location of viable and non-viable foetuses, early and late resorptions, total implantations and corpora lutea were recorded.

Statistics:

Statistical analysis were performed by using 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) to Differences in the foetal sex distribution and the number of litters with malformations between control and treated groups. 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 foetal weights were compared between groups by analysis of variance (one way classification), Bartlett's test for 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).

Clinical signs:

effects observed, treatment-related

Mortality:

mortality observed, treatment-related

Body weight and weight changes:

effects observed, treatment-related

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

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

effects observed, treatment-related

Histopathological findings: non-neoplastic:

not specified

Details on results:

Mortality :
When treated with 1500 mg/kg bw/day, Six rats died during the dosing period as compared to control.

Clinical signs:
Orange discoloration of the urine was observed in all the treated rats as compared to control.

Body weight and weight gain When treated with 1500 mg/kg bw/day, decrease in body weight gain was observed in treated rats as compared to control.

Food consumption and compound intake No data available

Food efficiency No data available

Water consumption and compound intake: No data available

Opthalmoscopic examination No data available

Haematology: No data available

Clinical chemistry: No data available

Urinanalysis: No data available

Neurobehaviour: No data available

Organ weights No data available

Gross pathology: When treated with 1500 mg/kg bw/day, green discoloration of the amniotic fluid and green colored small intestines were observed in treated rats as compared to control.

When treated with 100 and 500 mg/kg bw/day, green discoloration of the amniotic fluid was observed in treated rats as compared to control.

Histopathology No data available

Details on results:
There were no biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations, number of foetuses or litters with developmental variations in any of the treated groups.

At 1500 mg/kg be/day, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) was observed as compared to control. However, these values fell within the ranges of historical control data.

Dose descriptor:

NOAEL

Effect level:

500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: No effect on survival, clinical sign, body weight and body weight gain, gross pathology and reproductive performances

Critical effects observed:

not specified

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

		Mean maternal body weights (g)
Day of gestation	Dose (mg kg / day)	0 (control)	100	500	1500
0		246± 145	249 ± I3.7	249± 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

 

Mean maternal body-weight changes in rats given D & C Yellow No. 8 on days 6 -19 of gestation

		Mean maternal body weights (g)
Day of gestation	Dose (mg kg / day)	0 (control)	100	500	1500
0 -6		26	22	27	22
6 -9		7	7	7	6
9 -12		15	14	13	13
12 -16		27	28	27	18
16 -20		58	58	51	59
6-20		107	107	98	06
0-20		133	129	125	118

Conclusions:

NOAEL was considered to be 500 mg/kg body weight /day when CD Sprague-Dawley female rats were treated with D&C YELLOW NO. 8.

Executive summary:

In a Teratogenic toxicity study, CD Sprague-Dawley female rats were 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 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. In addition, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) were observed as compared to control. However, these values fell within the ranges of historical control data. There were no biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations,number of foetuses or litters with developmental variations in any of the treated groups. Therefore, NOAEL was considered to be 500 mg/kg body weight /day when CD Sprague-Dawley female rats were treated with D&C YELLOW NO. 8 orally by gavage from day 6 to 19 of gestation.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: Oral

Peer reviewed publications were assessed to determine the toxic nature of the test compound D & C Yellow No. 8 (CAS no 518-47-8). The summary is as mentioned below:

In a Teratogenic toxicity study conducted by Burneti et al(1986), CD Sprague-Dawley female rats were treated with D&C Yellow No. 8 (CAS no 518-47-8) in the concentration of 0, 100, 500 and 1500 mg/kg body weight/ day by oral gavage. Six rats died during the dosing period 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. In addition, slight increase in the number of litters with unossified sternebrae (sternebrae nos 1-6) and rudimentary 14th rib(s) were observed as compared to control. However, these values fell within the ranges of historical control data. There were no biologically meaningful or statistically significant differences in the number of litters and number of fetuses with malformations,number of foetuses or litters with developmental variations in any of the treated groups. Therefore, NOAEL was considered to be 500 mg/kg body weight /day when CD Sprague-Dawley female rats were treated with D&C Yellow No. 8 orally by gavage from day 6 to 19 of gestation.

In the same study, Dutch Belted female Rabbits were treated with D&C Yellow No. 8 (CAS no 518-47-8) in the concentration of 0, 30, 100 and 250 mg/kg body weight/ day by oral gavage. One rabbit died each in 30 and 250 mg/kg bw/day including control. Orange discoloration of the urine was observed in all the treated rabbits between days 7 and 28 of gestation as compared to control. In addition, no effects were observed on body weight and body weight gain and gross pathology of treated rabbits as compared to control. There were no biologically meaningful or statistically significant differences in the mean numbers of corpora lutea, total implantations+ early or late resorptions, viable foetuses, foetal sex distribution or mean foetal body weight in any of the treated groups as compared to the control group. At 30 and 100 mg/kg bw/day, malformations were observed in two foetuses from two litters. However, these values fell within the ranges of historical control data. Therefore, NOAEL was considered to be 250 mg/kg body weight /day when Dutch Belted female Rabbits were treated with D&C Yellow No. 8 orally by gavage from day 6 to 27 of gestation.

Based on the available key study and its supporting data, the test chemical D & C yellow no 8 is likely to be not toxic upon repeated exposure by the oral route.

Justification for classification or non-classification

Based on the studies summarized, the test chemical D & C yellow no 8 (CAS no 518 -47 -8) is likely to be not toxic upon repeated exposure by the oral route.