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

Genetic toxicity: in vivo

Administrative data

Endpoint:

in vivo mammalian cell study: DNA damage and/or repair

Remarks:

Type of genotoxicity: DNA damage and/or repair

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Data source

Reference

Reference Type:

publication

Title:

Testing of 24 Foods, Drug, Cosmetic, and Fabric Dyes in the In Vitro and the In Vivo/ In Vitro Rat Hepatocyte Primary Culture/ DNA Repair Assays

Author:

Douglas Kornbrust and Thomas Barfknecht

Year:

1985

Bibliographic source:

Environmental Mutagenesis 7:101-120 (1985)

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: as mentioned below

Principles of method if other than guideline:

DNA damage and/or repair toxicity performed on Sprague Dawley cesarean-derived rats

GLP compliance:

no

Type of assay:

other: In Vivo/ In Vitro Rat Hepatocyte Primary Culture/ DNA Repair Assays

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:

Food black

IUPAC Name:

Food black

Details on test material:

- Name of test material (as cited in study report): Food black (2519-30-4)- Molecular formula (if other than submission substance): Not applicable- Molecular weight (if other than submission substance): Not applicable- Substance type: organic - Physical state: solid- Purity: 95%- Impurities (identity and concentrations):5%

Test animals

Species:

rat

Strain:

Sprague-Dawley

Sex:

male

Details on test animals or test system and environmental conditions:

Details on test animals and env conditionsTEST ANIMALS- Source: No data available - Age at study initiation: No data available- Weight at study initiation: 200-300 g- Assigned to test groups randomly: [no/yes, under following basis: ] No data available- Fasting period before study: No data available- Housing: No data available- Diet (e.g. ad libitum): Diet (ad libitum)- Water (e.g. ad libitum): Water (ad libitum)- Acclimation period: No data availableENVIRONMENTAL CONDITIONS- Temperature (°C): No data available- Humidity (%):12-hr light/dark cycles- Air changes (per hr): No data available- Photoperiod (hrs dark / hrs light): 12-hr light/dark cyclesIN-LIFE DATES: From: To: No data available

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

Aqueous solution Or corn oil

Details on exposure:

No data available

Duration of treatment / exposure:

At specific times prior to the isolation of hepatocytes

Frequency of treatment:

One

Post exposure period:

No data available

Doses / concentrations

Remarks:

Doses / Concentrations:500 mg/kgBasis:no data

No. of animals per sex per dose:

No data available

Positive control(s):

Solvent Yellow 3 (0-aminoazotoluene)

Examinations

Tissues and cell types examined:

Rat hepatocytes

Details of tissue and slide preparation:

No data available

Evaluation criteria:

Net nuclear grains (NNG) were determined by counting the number of grains in each nuclei and subtracting the average number of grains present in three equal-size adjacent cytoplasmic areas.Net nuclear grains (NNG) were measured are as follows:Average net nuclear grain counts of 5 or greater were assumed to constitute a positive response. For those dyes that produced responses between zero and 5 average net nuclear grains, it was generally not possible to demonstrate a statistically significant difference from the control value within a given experiment. Therefore, these responses were judged to be equivocal. Net nuclear grain counts below zero were considered negative responses.

Statistics:

No data available

Results and discussion

Test results

Sex:

male

Genotoxicity:

negative

Toxicity:

not specified

Vehicle controls validity:

not specified

Negative controls validity:

not specified

Positive controls validity:

not specified

Any other information on results incl. tables

- Statistical evaluation:

 

Responses Produced by Various Dyes in the In Vivo/In Vitro HPC/DR Assay

Dye	Dose (mg/kg b.w.)	Time PTP^a (hr)	Avg. NNG^b	Percent cells<5 NNG	Res- ponse
Food Black 1	500	2 15	-0.7 (±3.3) -1.5 (± 2.8)	5 3	N
^aTime that dye was administered prior to start of liver perfusion and isolation of hepatocytes. ^bAverage net nuclear grains (as defined in the Methods section); mean + standard deviation from 60 cells. ^CPercent of cells with > 5 net nuclear grains. ^dP. Positive; WP, weak positive; E, equivocal; N, negative (as per the criteria defined in the methods section).

Applicant's summary and conclusion

Conclusions:

Interpretation of results (migrated information): negativeThe genetic toxicity of Food black 1 (CAS No 2519-30-4) in Rat hepatocytes cell line was found to be negative.

Executive summary:

The genetic toxicity test was performed on rat using Food black 1.Rats given the 500 mg/kg ofFood black 1.Dye were administered to the animals as aqueous solutions or corn oil suspensions (for the non-water-soluble dyes), by gavage, at specific times prior to the isolation of hepatocytes.

Rat hepatocytes were isolated and cultured by the two-step in situ liver perfusion

Method.2 X 10⁵viable hepatocytes were seeded into 25-mm wells and were allowed to attach to plastic cover slips for 2 hr.The cells were then incubated for 4 hr with [³H]-thymidine.

Net nuclear grains (NNG) were determined by counting the number of grains in each nuclei and subtracting the average number of grains present in three equal-size adjacent cytoplasmic areas.

Average net nuclear grain counts of 5 or greater were assumed to constitute apositiveresponse. For those dyes that produced responses between zero and 5 average net nuclear grains, it was generally not possible to demonstrate a statistically significant difference from the control value within a given experiment. Therefore, these responses were judged to beequivocal. Net nuclear grain counts below zero were considerednegativeresponses.

From the above table it was found that Avg. NNG forFood black 1 was found to be below zero.

 

Therefore, from the study in vivogenetic toxicity of Food black 1 (CAS No 2519-45-9)was found to be negative.