4,4'-[1,2-ethenediylbis[(3-sulfo-4,1-phenylene)imino[6-[bis(2-hydroxyethyl)amino]-1,3,5-triazine-4,2-diyl]imino]]bisarenoate sodium salt

Repeated dose toxicity: oral

Administrative data

Endpoint:

chronic toxicity: oral

Remarks:

combined repeated dose and carcinogenicity

Type of information:

migrated information: read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented study report which meets basic scientific principles

Cross-reference

Reason / purpose for cross-reference:

reference to same study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1978

Report date:

1978

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 453 (Combined Chronic Toxicity / Carcinogenicity Studies)

Deviations:

yes

Remarks:

(limited parameters examined)

GLP compliance:

no

Limit test:

no

Test material

Test material information

Constituent 1

Details on test material:

- Name of test substance (as cited in study report): Blankophor P (acidic sodiumsalt)
- Analytical purity: 81%
- Physical state: solid

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Winkelmann, Borchen, Germany
- Age at study initiation: 28 - 32 days old
- Weight at study initiation: mean: 48 g
- Housing: single
- Diet: Altromin R-Pulverfutter (Altromin GmbH, Lage/Lippe, Germany), ad libitum
- Water: tap water; ad libitum

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 ± 1

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

24 months

Frequency of treatment:

daily

Doses / concentrations

Doses / concentrations 1

Remarks:

Doses / Concentrations:
males: 5.23, 52.29, and 520.78 mg/kg bw/day; females: 7.02, 69.33, and 709.25 mg/kg bw/day
Basis:
actual ingested

Doses / concentrations 2

Remarks:

Doses / Concentrations:
100, 1000 and 10000 ppm
Basis:
nominal in diet

No. of animals per sex per dose:

- test groups: 50
- control group: 100

Control animals:

yes, plain diet

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No

BODY WEIGHT: Yes
- Time schedule for examinations: first 6 months weekly, after that every two weeks

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: No
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Yes

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): No

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at 1, 3, 6, 12 and 24 months
- Anaesthetic used for blood collection: No data
- Animals fasted: No data
- How many animals: 10/sex at 24 months otherwise 5/sex
- Parameters checked: haemoglobin, haematocrit, erythrocytes, leucocytes, MCV, MCH, reticulocyte, thromocytes

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at 1, 3, 6, 12 and 24 months
- Animals fasted: No data
- How many animals: 10/sex at 24 months otherwise 5/sex
- Parameters checked: ALP, GOT, GTP, bilirubin, protein, blood glucose, cholesterol

URINALYSIS: Yes
- Time schedule for collection of urine: at 1, 3, 6, 12 and 24 months (urine from 16 o'clock)
- Metabolism cages used for collection of urine: No data
- Animals fasted: No data
- Parameters checked: glucose, protein, blood, pH, ketone bodies, bile pigments, sediments, urea, creatinine

NEUROBEHAVIOURAL EXAMINATION: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes (aorta, eyes, duodenum, jejunum, urinary bladder, heart, testes, pituitary, liver, lung, lymph nodes, stomach, spleen, epididymis, adrenals, kidneys, os femoris, oesophagus, ovaries, pancreas, prostate, seminal vesicle, thymus, skeletal muscle, sternum, trachea, and uterus)

Statistics:

U-test (Mann); Whitney wilcoxon

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Clinical biochemistry findings:

no effects observed

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

no effects observed

Histopathological findings: neoplastic:

no effects observed

Details on results:

CLINICAL SIGNS AND MORTALITY: No differences compared to negative controls were observed.
- after one year: 2 male and 0 female of 100 animals died in the control group; at 100 ppm 1/50 males and 1/50 females died; at 1000ppm 3/50 males and 0/50 females and at 10000ppm 0/50 males and 0/50 females died
- after two years: in the control group 37/100 males and 25/100 females died; at 100 ppm 23/50 males and 17/50 females died; at 1000ppm 25/50 males and 14/50 females died and at 10000ppm 16/50 males and 17/50 females died

BODY WEIGHT AND WEIGHT GAIN: comparable in all groups

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study): normal in all groups

HAEMATOLOGY: No substance-related changes in any parameter were observed.

CLINICAL CHEMISTRY: after 1 month: increase in GOT and GPT at 10000 ppm (males), increase in GPT at 1000 and 10000 ppm (females); no dose-dependent changes in any parameter after 3, 6, 12 and 24 months treatment.

URINALYSIS: normal in all groups

ORGAN WEIGHTS: significant increase of kidey weight in males and females at 10000 ppm, as the difference is < 10%, it is not considered toxicologically relevant; the other changes in organ weight (liver, lung, heart, and testes) were considered not to be treatment-related as no dose-relationship was observed.

GROSS PATHOLOGY: no abnormalities detected

HISTOPATHOLOGY: no abnormalities detected; only findings that are age-related and common for this strain were observed

OTHER: From type, localisation and frequency of observed tumors there is no indication for cancerogenic effect of the test substance

Effect levels

1

Dose descriptor:

NOAEL

Effect level:

10 000 ppm

Based on:

test mat.

Sex:

male/female

Basis for effect level:

other: overall effects, highest dose tested

2

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

female

Basis for effect level:

other: overall effects, highest dose tested

3

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

other: overall effects, highest dose tested

Target system / organ toxicity

Critical effects observed:

not specified

Any other information on results incl. tables

Food and active ingredient intake:

Dosis [ppm]	mean food intake		mean active ingredient intake
	kg/animal	g/animal/day	g/kg bw	mg/kg bw /day
	male
0	16.48	22.43	-	-
100	16.49	22.44	3.84	5.23
1000	16.44	22.36	38.4	52.24
10000	16.61	22.6	382.77	520.78
	female
0	12.5	17.01	-	-
100	12.54	17.06	5.16	7.02
1000	12.79	17.4	50.96	69.33
10000	12.86	17.52	521.3	709.25

Mean absolute organ weights (mg)
Dosis (ppm)	Body weight (g)	Thymus	Heart	Lungs	Liver	Spleen	Kidneys	Adrenals	Testes/ ovary
Males
0	396	25	1246	2036	14364	742	2792	54	3331
100	372	26	1303	2716**	13036	665	2782	57	2908*
1000	368*	23	1184	2235	12755**	697	2740	51	3171
10000	390	27	1353**	1809	15524	790	3037**	55	3554
Females
0	264	21	959	1529	9507	530	1978	72	114
100	244*	19	947	1508	8854	551	1921	71	117
1000	267	19	971	1436	9600	608	2009	69	106
10000	265	20	973	1210*	9938	595	2108**	70	114

* significant different from control; p < 0.05

** significant different from control; p < 0.01

 

Applicant's summary and conclusion

Conclusions:

In this Combined Chronic Toxicity / Carcinogenicity Studies performed to a protocol similar to the OECD Guideline 453 no specific adverse effect was observed by treatment of rats with the test substance for 2 years.