Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate

Repeated dose toxicity: dermal

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

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

Test material information

Constituent 1

Reference substance name:

Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate

EC Number:

219-746-5

EC Name:

Tetrasodium 1-acetamido-2-hydroxy-3-(4-((4-sulphonatophenylazo)-7-sulphonato-1-naphthylazo))naphthalene-4,6-disulphonate

Cas Number:

2519-30-4

Molecular formula:

C28H21N5O14S4.4Na

IUPAC Name:

tetrasodium 4-acetamido-5-hydroxy-6-({7-sulfonato-4-[(4-sulfonatophenyl)diazenyl]-1-naphthyl}diazenyl)naphthalene-1,7-disulfonate

Constituent 2

Reference substance name:

Black PN

IUPAC Name:

Black PN

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 or test system 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.