Registration Dossier

Administrative data

Endpoint:
sub-chronic toxicity: dermal
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)

Data source

Reference
Reference Type:
publication
Title:
Physical Factors Determining the Early Local Tissue Reactions Produced by Food Colourings and Other Compounds Injected Subcutaneously
Author:
S. D. GANGOLLI, P. GRASSO and L. GOLBERG *
Year:
1967
Bibliographic source:
Fd Cosmet. Toxicol. Vol. 5, pp. 601-621, 1967

Materials and methods

Test guideline
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 411 (Subchronic Dermal Toxicity: 90-Day Study)
Principles of method if other than guideline:
Subcutaneous repeated dose toxicity study of Black PN in rats
GLP compliance:
not specified
Limit test:
no

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report):Black PN - Molecular formula (if other than submission substance):C28-H21-N5-O14-S4.4Na- Molecular weight (if other than submission substance):867.6873 g/mole- Substance type:Organic- Physical state:- Impurities (identity and concentrations):10 % Pb and 3 % As

Test animals

Species:
rat
Strain:
other: Shell or Carworth Farm E SPF
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS- Source: No data available - Age at study initiation: No data available - Weight at study initiation: 150- 300 g- Fasting period before study: No data available - Housing: No data available - Diet (e.g. ad libitum): Spillers Small Laboratory Animal diet, ad libitum- Water (e.g. ad libitum): Water, ad libitum- 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 availableIN-LIFE DATES: From: To: No data available

Administration / exposure

Type of coverage:
not specified
Vehicle:
not specified
Details on exposure:
PREPARATION OF DOSING SOLUTIONS: 1 ml of a 2 % aqueous solution of the test substance (pH 6.5-7.5) was injected subcutaneously. TEST SITE- Area of exposure: No data available- % coverage: No data available - Type of wrap if used: No data available - Time intervals for shavings or clipplings: No data available REMOVAL OF TEST SUBSTANCE- Washing (if done): No data available - Time after start of exposure: No data available TEST MATERIAL- Amount(s) applied (volume or weight with unit): No data available - Concentration (if solution): 1 ml - Constant volume or concentration used: yes- For solids, paste formed: noVEHICLE- Justification for use and choice of vehicle (if other than water): No data available - Amount(s) applied (volume or weight with unit): No data available- Concentration (if solution): 2000 mg/kg- Lot/batch no. (if required): No data available- Purity: No data availableUSE OF RESTRAINERS FOR PREVENTING INGESTION: No data available
Analytical verification of doses or concentrations:
not specified
Duration of treatment / exposure:
10 weeks
Frequency of treatment:
Twice-weekly
Doses / concentrations
Remarks:
Doses / Concentrations:2000 mg/kg Basis:nominal per unit area
No. of animals per sex per dose:
No data available
Control animals:
not specified
Details on study design:
No data available
Positive control:
No data available

Examinations

Observations and examinations performed and frequency:
No data available
Sacrifice and pathology:
GROSS PATHOLOGY: No data availableHISTOPATHOLOGY: Yes The skin and subcutaneous tissue were removed, fixed in buffered formol saline and prepared for histological examination. Rates of absorption, Relationship between surface activity and tissue reaction and Protein-binding ability and lipophilic properties of protein-colouring complexes were examined.
Other examinations:
No data available
Statistics:
No data available

Results and discussion

Results of examinations

Clinical signs:
not specified
Dermal irritation:
not specified
Mortality:
not specified
Body weight and weight changes:
not specified
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:
not specified
Histopathological findings: non-neoplastic:
no effects observed
Details on results:
Clinical signs and mortalityNo data availableDermal IrritationNo data availableBody weight and weight gainNo data availableFood consumption and compound intakeNo data availableFood efficiencyNo data availableWater consumption and compound intakeNo data availableOpthalmoscopic examinationNo data availableHaematologyNo data availableClinical chemistryNo data availableUrinanalysisNo data availableNeurobehaviourNo data availableOrgan weightsNo data availableGross pathologyNo data availableHistopathologyWhen treated with 2000 mg/kg, Type ll tissue reactions were observed i.e. insufficient progressive lesions in treated rats. Details on resultsType II are not associated with the reported development of sarcoma in long-term tests

Effect levels

Dose descriptor:
NOAEL
Effect level:
2 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No effect on histopathology

Target system / organ toxicity

Critical effects observed:
not specified

Any other information on results incl. tables

Surface activity, lipid solubility and protein binding ability of food colouring studies

 

Lipophilic properties

Injected material

type of reaction after 10-20 injections

Depression of surface tension of water (%)

Protein binding (g/100 g protine)

colouring combining with 100 mM cholesterol lecithin mixed miceller solution (mM)

Protein-colouring complex partition in tricaprylan (%)

Colouring

Cation

Concen (%)

Vol. (ml)

 

 

 

 

 

 

 

 

 

Azo

Black PN

Na

3

1.0

ll

6.3

96

4.8

0.1

 

 

 

 

 

 

 

 

 

Applicant's summary and conclusion

Conclusions:
NOAEL was considered to be 2000 mg/kg when Shell or Carworth Farm E SPF male and female rats were treated with Black PN
Executive summary:

In a Subcutaneous repeated dose toxicity study, Shell or Carworth Farm E SPF male and female rats were treated with Black PN in the concentration of 2000 mg/kg Twice-weekly. Skin and subcutaneous tissue were examined histopathologically. Type ll tissue reactions were observed i.e. insufficient progressive lesions in treated rats. In addition, Black PN shows weak physicochemical properties which indicate a self-limiting reaction (type II). Therefore, NOAEL was considered to be 2000 mg/kg when Shell or Carworth Farm E SPF male and female rats were treated with Black PN Subcutaneous for 10 weeks.