Trisodium 8-hydroxypyrene-1,3,6-trisulphonate

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Dec 1976 to March 1977

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

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 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

GLP compliance:

no

Remarks:

Study conducted prior to implementation of OECD GLP

Test material

Test material information

Constituent 1

Reference substance name:

Trisodium 8-hydroxypyrene-1,3,6-trisulphonate

EC Number:

228-783-6

EC Name:

Trisodium 8-hydroxypyrene-1,3,6-trisulphonate

Cas Number:

6358-69-6

Molecular formula:

C16H10O10S3.3Na

IUPAC Name:

trisodium 8-hydroxypyrene-1,3,6-trisulphonate

Test animals

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Breeder Winkelmann, Borchen
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 32 - 35 days
- Weight at study initiation: males 87 - 92 g / females 80 - 82 g
- Fasting period before study: no information given
- Housing: individually in Macrolon type II cages
- Diet: ad libitum
- Water: ad libitum


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 23 ± 2°C

Administration / exposure

Route of administration:

oral: feed

Vehicle:

unchanged (no vehicle)

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet (frequency): weekly
- Mixing appropriate amounts with (Type of food): Altromin-R-powder

Analytical verification of doses or concentrations:

not specified

Duration of treatment / exposure:

3 months

Frequency of treatment:

continuous

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 ppm

Doses / concentrations 2

Dose / conc.:

100 ppm

Remarks:

8.34 mg/kg bw/day (males); 12.28 mg/kg bw/day (females)
calculated based on mean weekly food consumtion relating to mean body weight (week 15)

Doses / concentrations 3

Dose / conc.:

1 000 ppm

Remarks:

81.40 mg/kg bw/day (males); 121.32 mg/kg bw/day (females)
calculated based on mean weekly food consumtion relating to mean body weight (week 15)

Doses / concentrations 4

Dose / conc.:

10 000 ppm

Remarks:

849.28 mg/kg bw/day (males); 1245.05 mg/kg bw/day (females)
calculated based on mean weekly food consumtion relating to mean body weight (week 15)

No. of animals per sex per dose:

15

Control animals:

yes, concurrent no treatment

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: weekly

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: Yes
- 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

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: after 4 weeks and at termination
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: 5 males/5 females
- Parameters examined: haematocrit, haemoglobin concentration, erythrocyte count, MCH and MCV, total and differential leukocyte count, platelet count, and a measure of blood clotting time/potential.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: after 4 weeks and at termination
- Animals fasted: Not specified
- How many animals: 5 males/5 females
- Parameters examined: alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, bilirubin, total protein, glucose, total cholesterol,

URINALYSIS: Yes
- Time schedule for collection of urine: after 4 weeks and at termination
- Metabolism cages used for collection of urine: Not specified
- Animals fasted: Not specified
- Parameters examined: glucose, ketone bodies, bilirubin, total protein, urobilinogen, pH-value, sediment, urea, creatinine

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, treatment-related

Description (incidence and severity):

slight reduction of body weight development for male rats in the highest dose group

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

slight but significant and dose dependent increase in thromboplastin time for male rats in the mid and high dose groups.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

slight decrease in bilirubin values for female rats in the high dose group after 4 weeks (no changes at termination).

Urinalysis findings:

no effects observed

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, non-treatment-related

Description (incidence and severity):

- male rats showed significant and dose dependant lower absolute heart weights in all dose groups.
- male rats showed slight but significant lower testis weights in the high dose group

Gross pathological findings:

no effects observed

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

sporadic findings in individual animals were:
- indications of inflamation in lung and liver (not specified)
- round cell infiltration in myocard of and in kindney one control rat respecitvely
- dilation of uterus (individual control and treated animals)
- atrophy of testis (one male high dose group)

Histopathological findings: neoplastic:

no effects observed

Other effects:

not specified

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

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

Based on:

test mat.

Sex:

male

Basis for effect level:

body weight and weight gain

2

Key result

Dose descriptor:

NOAEL

Effect level:

1 245.05 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: highest dose tested without adverse effects

Target system / organ toxicity

Key result

Critical effects observed:

no

Any other information on results incl. tables

Male and female rats received pyranin doses of 0, 100, 1000 and 10,000 ppm, respectively, administered with the diet.

The animals fed in this way showed no treatment-related symptoms in any dose group during the entire experimental period. The growth of male rats in the dose groups up to 1000 ppm as well as the female rats of all treatment groups was unaffected. In the male rats, 10000 ppm resulted in a slight growth retardation in the second half of the experiment. There were no mortalities.

 

The significant and dose-dependent prolongation of thromboplastin concentrations in male rats of the experimental groups dosed with 1000 and 10.000 ppm in the blood test after 3 months are not considered to be an expression of a disturbance in the coagulation system since the differences to the control group are very small and all values were within the normal range for this rat strain. Thus the blood was not damaged by the treatment.

 

In the female animals of the high dose group group, the bilirubin values measured 4 weeks after the start of the experiment were also within the normal historical range (2s range 0.10 to 0.26 mg / 100 ml measured in 100 untreated female rats of the same age). Clinical-chemical analyzes, gross pathology, and histopathological examinations did not reveal any signs of treatment-induced liver damage.

 

Urine tests, concentrations of urea and creatinine in the serum as well as macroscopic and histopathological organ findings did not indicate any influence on the kidneys.

 

Blood glucose and cholesterol levels were not changed during treatment or at termination.

No treatment related changes were found in the gross pathological examination of all rats. The significantly lower cardiac and test weights in the male rats corresponded approximately to the somewhat lower body weights of the treatment groups and because there was no histopathological corelate they were not regarded as toxicologically relevant.

 

The results obtained during the histopathological examination do not exceed the usual extent for conventional rats and are not due to treatment with pyranine.

 

Doses up to 1000 ppm (81.4 mg/kg bw/day) were thus tolerated by the male rats, doses up to 10,000 ppm (1245.05 mg/kg bw/day) by the female rats without any adverse effects.

Applicant's summary and conclusion

Conclusions:

Doses up to 1000 ppm (81.4 mg/kg bw/day) were tolerated by the male rats, doses up to 10,000 ppm (1245.05 mg/kg bw/day) were tolerated by the female rats without any adverse effects.

Executive summary:

Groups of 15 male and 15 female rats and 30 animals in each of the control groups received trisodium 8-hydroxypyrene-1,3,6-trisulphonate for 3 months administered with the diet in the following concentrations: 0 (control), 100, 1000, and 10,000 ppm.

 

Appearance, behavior, and mortality were unaffected in male and female rats receiving doses up to 1000 ppm. Doses up to 1000 ppm (81.4 mg/kg bw/day) in the male rats or up to 10,000 ppm (1245.05 mg/kg bw/day) in the female rats had no effect on the body weight gain.

In the highest dose group receiving 10,000 ppm (849.28 mg/kg bw/day) the body weight gain of the male rats was slightly delayed.

 

The blood was not damaged in the dose groups up to 10,000 ppm (849.28 mg/kg bw/day males/ 1245.05 mg/kg bw/day females).

 

According to the clinico-chemical, macroscopic-anatomical and histopathological studies, the male and female rats dosed with up to 10,000 ppm (849.28 mg/kg bw/day males/ 1245.05 mg/kg bw/day females) did not show any evidence for liver damage.

 

Urine tests, clinical-chemical analyzes, gross - and histopathological examinations did not provide any evidence for renal impairment in dose groups up to 10,000 ppm (849.28 mg/kg bw/day males/ 1245.05 mg/kg bw/day females).

Blood glucose and cholesterol concentrations were in the normal range in male and female rats up to the dose groups of 10,000 ppm (849.28 mg/kg bw/day males/ 1245.05 mg/kg bw/day females).

 

Gross pathology and histopathological examinations did not give any indications for treatment-related organ changes in dose groups up to 10,000 ppm (849.28 mg/kg bw/day males/ 1245.05 mg/kg bw/day females).

 

Doses of up to 1000 ppm (81.4 mg/kg bw/day) for male rats and up 10,000 ppm (1245.05 mg/kg bw/day) for female rats were tolerated without any effects under the conditions described in this study.