3-(2,4-bis(4-((5-(4,6-bis(2-aminopropylamino)-1,3,5-triazin-2-ylamino)-4-hydroxy-2,7-disulfonaphthalen-3-yl)azo)phenylamino)-1,3,5-triazin-6-ylamino)propyldiethylammonium lactate

Endpoint summary

Description of key information

NOAEL >= 1000 mg/kg bw/day

Key value for chemical safety assessment

Toxic effect type:

dose-dependent

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

short-term repeated dose toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From May 15th to July 3rd, 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

EU Method B.7 (Repeated Dose (28 Days) Toxicity (Oral))

Version / remarks:

July 31st, 1992

Qualifier:

according to guideline

Guideline:

OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity Study in Rodents)

Version / remarks:

May 12th, 1981

GLP compliance:

yes (incl. QA statement)

Limit test:

no

Specific details on test material used for the study:

Stable in polyethylene glycol and water fo 48 hours.

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

Total number of animals: 30 males, 30 females
Total number of animals per group: Groups 1 and 4: 10 males, 10 females; Groups 2 and 3: 5 males and 5 females

Route of administration:

oral: gavage

Vehicle:

other: PEG400 / bi-distilled water 1:1

Analytical verification of doses or concentrations:

yes

Duration of treatment / exposure:

Duration of acclimatization period: 7 days
Duration of treatment: 28 days
Duration of recovery: 14 days

Frequency of treatment:

Dosing regime: 7 days/week

Dose / conc.:

0 mg/kg bw/day (nominal)

Remarks:

Control

Dose / conc.:

50 mg/kg bw/day (nominal)

Dose / conc.:

200 mg/kg bw/day (nominal)

Dose / conc.:

1 000 mg/kg bw/day (nominal)

No. of animals per sex per dose:

Male: 10 animals at 0 mg/kg bw/day
Male: 5 animals at 50 mg/kg bw/day
Male: 5 animals at 200 mg/kg bw/day
Male: 10 animals at 1000 mg/kg bw/day

Female: 10 animals at 0 mg/Kg bw/day
Female: 5 animals at 50 mg/Kg bw/day
Female: 5 animals at 200 mg/Kg bw/day
Female: 10 animals at 1000 mg/Kg bw/day

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

Clinical signs, food consumption and body weights were recorded periodically during the treatment and recovery periods. Ophthalmoscopic examinations were performed at the end of the treatment ad recovery periods.
At the end of the dosing period and the treatment-free period blood samples were withdrawn for haematology and plasma chemistry analysis, and urine samples were collected for biochemical and microscopic analyses. All animals were killed, examine post-mortem and necropsied. Samples of major organs from all group 0 mg/kg and 1000 mg/kg animals and gross lesions from all animals were processed as haematoxylin and eosin stained slides and examine by light microscopy.

Clinical signs:

no effects observed

Description (incidence and severity):

There were no test substance-related clinical signs note in any group.
In one female treated with 100 mg/kg dose rales were noted on treatment days 11-12. No further clinical signs were observed in any other animal treated at 1000 mg/kg, therefore the rales were considered of no toxicological significance.

Description (incidence):

All animals survived the scheduled study period. One control female and one female treated with 1000 mg/kg dose died after blood sampling on the day of the scheduled necropsy after 4 weeks of treatment.

Body weight and weight changes:

no effects observed

Food efficiency:

no effects observed

Ophthalmological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Ophthalmologic findings were noted in small proportion of animals from all groups. They included persistent pupillary membrane, corneal opacity, anterior synechia and vitrous floaters. These findings occurred at similar incidences in the control and treated groups at the end of the treatment period. Therefore, they are considered to be unrelated to treatment with the test substance.

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The only change of note was a slightly increate uric acid, total bilirubin, sodium, total protein and globulin level, and a slightly decreased chloride level in males treated with 1000 mg/kg at termination of the treatment. These findings, which were of minor degree and within historical control data, suggest metabolic adaptive changes rather than to constitute toxic effects. At the termination of the treatment-free recovery period these findings were generally reversed and found to be similar to those of the controls. All other statistical differences in the results of the haematology, clinical biochemistry and urinalysis data were considered to be incidental and unrelated to the treatment, and of normal biological variation for rats of this strain and age data for untreated Wistar rats.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

see haematological findings.

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

see haematological findings.

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

Higher absolute and relative ovaries weights after 6 weeks were recorded in females at 1000 mg/kg. Both statistically significant findings were not considered to be test articles-related, but due to the slightly higher body weight of animals at 1000 mg/kg

Gross pathological findings:

effects observed, non-treatment-related

Description (incidence and severity):

The macroscopic findings recorded were unremarkable and within the range of spontaneous alterations which may be seen in rats of this age and strain. They included dilated renal pelves, reddish discoloration in various organs and diluted uterine horns. Bluish discoloration of the gastrointestinal tract was noted amongst treated animals and was considered due to passive coloration by the test substance.

Dose descriptor:

NOAEL

Effect level:

1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food efficiency

gross pathology

haematology

mortality

ophthalmological examination

organ weights and organ / body weight ratios

urinalysis

Dose descriptor:

NOEL

Effect level:

200 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Basis for effect level:

body weight and weight gain

clinical biochemistry

clinical signs

food efficiency

gross pathology

haematology

mortality

ophthalmological examination

organ weights and organ / body weight ratios

urinalysis

Critical effects observed:

no

Conclusions:

Based on the results of this study, 1000 mg/kg bw/day of the substance was established as the NOAEL and 200 mg/kg as the no-observed-effect-level NOEL.

Executive summary:

The subacute 28-Day oral toxicity study was conducted according to OECD 407 (1981) and EU Method B.7 (1992). 

Dose levels for a study of 28 days oral treatment were selected to be 0, 50, 200 and 1000 mg/kg bw/day.

In this subacute 28-day toxicity study, the test item was administered daily by gavage to SPF-bred Wistar rats, followed by a 14-day recovery period.

A NOEL was established at 200 mg/kg bw/day, and the NOAEL was established at 1000 mg/kg bw/day.

Based on findings, the test item is not classified for Repeated dose toxicity (oral) according to the CLP Regulation (EC) No.1272/2008.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subacute

Species:

rat

Additional information

The subacute 28-Day oral toxicity study was conducted according to OECD 407 (1981) and EU Method B.7 (1992). 

The test item was administered daily by gavage to SPF-bred Wistar rats, followed by a 14-day recovery period, at dose levels of 0, 50, 200 and 1000 mg/kg bw/day. 

No adverse effects were observed up to the highest dose level therefore the NOAEL was established at 1000 mg/kg bw/day.

Justification for classification or non-classification

Based on the experimental evidences in the assessment of the toxicity potential after repeated oral exposure, the test item does not need to be classified according to the CLP Regulation (EC 1272/2008).