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Toxicological information

Repeated dose toxicity: oral

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Administrative data

Endpoint:
short-term repeated dose toxicity: oral
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 is from peer reviewed journal

Data source

Reference
Reference Type:
publication
Title:
Absorption and elimination of D&C Red No. 28 in male F-344 rats
Author:
C.J. Sweeta, A.M. So´ lyoma, I.G. Sipesb,*
Year:
2004
Bibliographic source:
Food and Chemical Toxicology 42 (2004) 641–648

Materials and methods

Test guideline
Qualifier:
according to
Guideline:
other: Refer below principle
Principles of method if other than guideline:
Repeated dose oral toxicity study was performed to determine the toxic nature of D&C Red No. 28 using rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Test material form:
solid: particulate/powder
Remarks:
migrated information: powder
Details on test material:
- Name of test material: D&C Red No. 28
- Molecular formula: 20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mole
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 5 %
Specific details on test material used for the study:
- Name of test material: D&C Red No. 28
- IUPAC name: 2',4',5',7'-tetrabromo-4,5,6,7-tetrachloro-3',6'-dihydroxy-3H-spiro[2-benzofuran-1,9'-xanthen]-3-one
- Molecular formula: 20H2Br4Cl4O5Na2
- Molecular weight: 829.66 g/mole
- Substance type: Organic
- Physical state: Solid
- Impurities (identity and concentrations): 5 %

Test animals

Species:
rat
Strain:
Fischer 344
Details on species / strain selection:
No data
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Sprague–Dawley Inc. (Indianapolis, IN).
- Age at study initiation: 8-week-old
- Weight at study initiation: 168 ± 6 g
- Fasting period before study: No data available
- Housing: Animals were housed in individual Nalgene metabolism cages or wire hanging cages.
- Diet (e.g. ad libitum): Teklad 4% Mouse-Rat Diet, ad libitum
- Water (e.g. ad libitum): Water, ad libitum
- Acclimation period: 5–7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20 ± 23.33 °C
- Humidity (%): 40-60%
- Air changes (per hr): 15 fresh filtered air changes per hour
- Photoperiod (hrs dark / hrs light): 12 h light/dark cycle

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

Administration / exposure

Route of administration:
oral: feed
Details on route of administration:
No data
Vehicle:
other: Blended rat chow (Teklad 4% Mouse-Rat Diet)
Details on oral exposure:
PREPARATION OF DOSING SOLUTIONS: Diet was prepared by blending rat chow (Teklad 4% Mouse-Rat Diet) mixed with Red 28 for 14 days such that their daily intake would be 500 mg/kg.

DIET PREPARATION
- Rate of preparation of diet (frequency): Every three days
- Mixing appropriate amounts with (Type of food): Blending rat chow (Teklad 4% Mouse-Rat Diet)
- Storage temperature of food: No data available

VEHICLE
- Justification for use and choice of vehicle (if other than water): blending rat chow (Teklad 4% Mouse-Rat Diet)
- Concentration in vehicle: 500 mg/kg/day
- Amount of vehicle (if gavage): No data available
- Lot/batch no. (if required): No data available
- Purity: No data available
Analytical verification of doses or concentrations:
not specified
Details on analytical verification of doses or concentrations:
No data
Duration of treatment / exposure:
14 days
Frequency of treatment:
Daily
Doses / concentrations
Remarks:
Doses / Concentrations:
500 mg/kg/day
Basis:
nominal in diet
No. of animals per sex per dose:
500 mg/kg: Two groups each of 28 male rats
Control animals:
not specified
Details on study design:
Data not available
Positive control:
Data not available

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Data not available
- Time schedule: Data not available
- Cage side observations checked in table [No.?] were included. Data not available

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Data not available

BODY WEIGHT: Yes
- Time schedule for examinations: Every other day

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): Yes
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY: Data not 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: Data not available

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Data not available
- Time schedule for examinations: Data not available

OPHTHALMOSCOPIC EXAMINATION: Data not available
- Time schedule for examinations: Data not available
- Dose groups that were examined: Data not available

HAEMATOLOGY: Data not available
- Time schedule for collection of blood: Data not available
- Anaesthetic used for blood collection: Data not available
- Animals fasted: Data not available
- How many animals: Data not available
- Parameters checked in table [No.?] were examined. Data not available

CLINICAL CHEMISTRY: Data not available
- Time schedule for collection of blood: Data not available
- Animals fasted: Data not available
- How many animals: Data not available
- Parameters checked in table [No.?] were examined. Data not available

URINALYSIS: Yes
- Time schedule for collection of urine: Data not available
- Metabolism cages used for collection of urine: Data not available
- Animals fasted: Yes, for 24 hours
- Parameters checked in table [No.?] were examined. Presence of D&C Red No. 28 in urine were examined.

NEUROBEHAVIOURAL EXAMINATION: Data not available
- Time schedule for examinations: Data not available
- Dose groups that were examined: Data not available
- Battery of functions tested: sensory activity / grip strength / motor activity / other: Data not available

OTHER: Data not available
Sacrifice and pathology:
No data
Other examinations:
To determine plasma kinetics and background levels of Red 28 from the diet, four rats from each group were killed by CO2 inhalation at 1, 2, 4, 8, 12, 24 and 48 h post dosing.
Statistics:
No data

Results and discussion

Results of examinations

Clinical signs:
not specified
Mortality:
not specified
Body weight and weight changes:
no effects observed
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
Immunological findings:
not specified
Organ weight findings including organ / body weight ratios:
not specified
Gross pathological findings:
not specified
Neuropathological findings:
not specified
Histopathological findings: non-neoplastic:
not specified
Histopathological findings: neoplastic:
not specified
Other effects:
not specified
Details on results:
Mortality: No data

Clinical signs: No data

Body weight and weight gain : An increase in body weight gain was observed in treated rats over the 14 days study period. 8-week-old rats at the start of the study weiged 168 g±6 g and by 10 weeks they weighed 225 g±10 g.

As there is no control in the study, the effects were not supposed to be treatment related.

Food consumption: No data

Compound intake: Daily intake would be 500 mg/kg for 14 days
.
Food efficiency: No data

Water consumption and compound intake: No data

Opthalmoscopic examination: No data

Haematology : No data

Clinical chemistry: No data

Urinanalysis: No Red 28 dye was detected in the urine or cage rinse of treated rats.

Neurobehaviour: No data

Organ weights: No data

Gross pathology: No data

Histopathology: No data

Effect levels

Dose descriptor:
NOAEL
Effect level:
500 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: No effect on body weight

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Detectable plasma levels of Red 28 were not present in the animals that received the bolus dose of water following pretreatment with Red 28 in the diet. In the animals that received the bolus dose of Red 28, the average maximum concentration of Red 28 measured in the plasma was 2.2 nmol/ml (0.01% of the bolus dose in total plasma volume). Red 28 was not detected in plasma beyond 12 h post dosing. The terminal half-life was calculated to be 1.7 h with a terminal rate constant of 0.41 h-1. Oral bioavailability of Red 28 was determined to be 0.60%.

Following a 14-day pretreatment regimen with Red 28 in the diet, animals that received the bolus dose of the vehicle (i.e., no Red 28) on day fifteen still excreted dye in the feces 24 h after being placed on the control diet. The amount of Red 28 recovered was 24501020 nmol/ g feces. The amount of dye recovered in the feces from these animals during the next 24 h declined dramatically (4648 nmol/g). As occurred with naı¨ve animals administered a large oral bolus dose of Red 28, animals that had been pretreated for 14 days developed diarrhea when administered the large bolus dose (500 mg/kg) by oral gavage.

Applicant's summary and conclusion

Conclusions:
The No Observed Adverse Effdect level (NOAEL) was considered to be 500 mg/kg/day when Fischer-344 (F-344) male rats were treated with D&C Red No. 28
Executive summary:

Repeated dose oral toxicity study was performed to determine the toxic nature of D&C Red No. 28 using rats. Two groups of 8 week old 28 male Fischer-344 rats each were fed a diet containing blended rat chow (Teklad 4% Mouse-Rat Diet) mixed with Red 28 for 14 days such that their daily intake would be 500 mg/kg. Rats were weighed every other day. Food containers were also weighed every day to determine the amount of consumed diet. The diet was prepared and replaced every three days based on the actual food consumption and weight of the animals over the two-week period. On day 15, after a 12 h fasting period, one group was dosed by oral gavage with the dye (500 mg/kg; 2 ml/kg) in water while the other group was dosed with the vehicle (water, 2 ml/kg). To determine plasma kinetics and background levels of Red 28 from the diet, four rats from each group were killed by CO2 inhalation at 1, 2, 4, 8, 12, 24 and 48 h post dosing. At these times, blood (4–5 ml) was immediately collected from the posterior vena cava into a heparinized syringe and processed. An increase in body weight gain was observed in treated rats over the 14 days study period. 8-week-old rats at the start of the study weiged 168 g±6 g and by 10 weeks they weighed 225 g±10 g. As no control animals were included in the study, the effects were not supposed to be treatment related. No Red 28 dye was detected in the urine or cage rinse of treated rats. Based on the observations made, the No Observed Adverse Effdect level (NOAEL) was considered to be 500 mg/kg/day when Fischer-344 (F-344) male rats were treated with D&C Red No. 28.