Reaction mass of lithium sodium 5-amino-3-{[4-(2-{4-[(7-amino-1-hydroxy-3-sulfo-2-naphthyl)diazenyl]-2-sulfophenyl}vinyl)-3-sulfophenyl]diazenyl}-4-hydroxynaphthalene-2,7-disulfonate 2,2'-(methylimino)diethanol (1:1) and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[6-amino-4-hydroxynaphthalene-2-sulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol

Toxicity to reproduction: other studies

Administrative data

Endpoint:

toxicity to reproduction: other studies

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study and GLP

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2013

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD TG 407

GLP compliance:

yes

Type of method:

in vivo

Test material

Test material information

Constituent 1

Reference substance name:

Reaction mass of lithium sodium 5-amino-3-{[4-(2-{4-[(7-amino-1-hydroxy-3-sulfo-2-naphthyl)diazenyl]-2-sulfophenyl}vinyl)-3-sulfophenyl]diazenyl}-4-hydroxynaphthalene-2,7-disulfonate 2,2'-(methylimino)diethanol (1:1) and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[6-amino-4-hydroxynaphthalene-2-sulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol

EC Number:

916-916-7

Molecular formula:

not available

IUPAC Name:

Reaction mass of lithium sodium 5-amino-3-{[4-(2-{4-[(7-amino-1-hydroxy-3-sulfo-2-naphthyl)diazenyl]-2-sulfophenyl}vinyl)-3-sulfophenyl]diazenyl}-4-hydroxynaphthalene-2,7-disulfonate 2,2'-(methylimino)diethanol (1:1) and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[6-amino-4-hydroxynaphthalene-2-sulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol

Test material form:

other: liquid

Details on test material:

Dye content: 41.2 %

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Age at study initiation: 6-7 weeks
- Weight at study initiation (mean): males 193g , females 154 g
- Housing: in groups of 2 or 3 animals
- Diet ad libitum
- Water . ad libitum:
- Acclimation period: approximately 1 week

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22
- Humidity (%): 55
- Air changes (per hr): >10
- Photoperiod (hrs dark / hrs light): 12/12

Administration / exposure

Route of administration:

oral: gavage

Vehicle:

water

Details on exposure:

Bayscript Blaukomponente (konserviert) was adminestered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle in daily doses of 0, 100, 300, 1000 mg/kg bw/day for aperiod of four weeks

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Before the start of the treatment the suitabilits of the proposed formulations was confirmed by the analyses of condentration and stabilits
Stability: the dosage forms prepared were analysed 4 hoursm 1,4,and 7 days after preparation
Concentration: the concenntration of samples of control and each tes substance dosage form prepared were determined twice during the study

Duration of treatment / exposure:

males 28 days, females 29 days

Frequency of treatment:

once daily

Duration of test:

30 days

Doses / concentrations

Remarks:

Doses / Concentrations:
0, 100, 300, 1000 mg/kg bw/day
Basis:

No. of animals per sex per dose:

5

Control animals:

yes, concurrent vehicle

Details on study design:

Bayscript Blaukomponente (konserviert) was adminestered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle in daily doses of 0, 100, 300, 1000 mg/kg bw/day for aperiod of four weeks.The animals were regularely observed, weighed and food and water intakes were determined. O addition, clinical laboratory investigations of blood samples was performed. Organs and tissues wre subjected to gross and histopathological investigation.

Statistics:

Dunnett, U-test, Het-Dunn ( Dunnett exact test heterogeneous test)

Results and discussion

Effect levels

1

Dose descriptor:

other: NOAEL (general toxicity)

Effect level:

300 mg/kg bw/day (actual dose received)

Sex:

male/female

2

Dose descriptor:

other: NOAEL (reproductive organs)

Effect level:

1 000 mg/kg bw/day (actual dose received)

Sex:

male/female

Basis for effect level:

other: no relevant adverse effects observed

Observed effects

Bayscript Blaukomponente (konserviert) was administered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle, in daily doses of 0, 100, 300, 1000 mg/kg bw/day for a period of four weeks. This GLP study was performed according to OECD guideline 407.
Treatment with Bayscript Blaukomponente resulted in several findings which were related to the color of the test substance. The whole body and several organs of animals treated with 1000 mg/kg bw/day appeared discolored. The kidneys were also discolored in the lower dose groups.
In the kidneys, beneath findings associated with the color of the test substance (starting at 300 mg/kg b.w./day), corticotubular degeneration and increased incidence and/or severity of cortical basophilic (regenerative) tubules were noted at 1000 mg/kg b.w./day. Furthermore, demyelinated fibers in nerve roots at the spinal cord were seen in both sexes at 1000 mg/kg b.w./day.
At 1000 mg/kg b.w./day, histopathological findings in the mesenteric lymph node, the interstitium of the testis, and in the Peyer's patch were related to the colored test item and not accompanied by other structural changes of the organs.
Minor changes in red blood cell parameters observed for males and females at 1000 mg/kg b.w./day were associated with appearing of golden-brown pigment deposition in the spleen in females. In males, the thrombocyte count was increased at 1000 mg/kg b.w./day.

Despite this general toxic effects no pathological changes were observed at the reproductive organs in amle and female rats

Under the conditions described the no adverse observed effect level (NOAEL) for administration of BAYSCRIPT Blaukomponente (konserviert) to male and female Wistar rats was 300 mg/kg b.w./day mainly due to degenerative effects on kidneys and nerve roots. With respect to reproductive organ toxitcity a NOAEL of 1000 mg/kgbw/day is established for male and female rats

Applicant's summary and conclusion

Executive summary:

Bayscript Blaukomponente (konserviert) was administered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle, in daily doses of 0, 100, 300, 1000 mg/kg bw/day for a period of four weeks. This GLP study was performed according to OECD guideline 407.

Treatment with Bayscript Blaukomponente resulted in several findings which were related to the color of the test substance. The whole body and several organs of animals treated with 1000 mg/kg bw/day appeared discolored. The kidneys were also discolored in the lower dose groups.

In the kidneys, beneath findings associated with the color of the test substance (starting at 300 mg/kg b.w./day), corticotubular degeneration and increased incidence and/or severity of cortical basophilic (regenerative) tubules were noted at 1000 mg/kg b.w./day. Furthermore, demyelinated fibers in nerve roots at the spinal cord were seen in both sexes at 1000 mg/kg b.w./day.

At 1000 mg/kg b.w./day, histopathological findings in the mesenteric lymph node, the interstitium of the testis, and in the Peyer's patch were related to the colored test item and not accompanied by other structural changes of the organs.

Minor changes in red blood cell parameters observed for males and females at 1000 mg/kg b.w./day were associated with appearing of golden-brown pigment deposition in the spleen in females. In males, the thrombocyte count was increased at 1000 mg/kg b.w./day.

Despite this general toxic effects no pathological changes were observed at the reproductive organs in amle and female rats

Under the conditions described the no adverse observed effect level (NOAEL) for administration of BAYSCRIPT Blaukomponente (konserviert) to male and female Wistar rats was 300 mg/kg b.w./day mainly due to degenerative effects on kidneys and nerve roots. With respect to reproductive organ toxitcity a NOAEL of 1000 mg/kgbw/day is established for male and female rats