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Administrative data

Description of key information

Based on all pieces of available information it is clear that tributyl citrate is of very low  toxicity after repeated administrations.
1) A GLP test equivalent to OECD 408 resulted in a NOAEL (rat) of 1000 mg/kg bw . [Read-across data from tributyl-O-acetylcitrate (CAS 77-90-7)]
2) A GLP test according to 875/318/EEC, 83/571/EEC and 91/507/EEC resulted in a NOAEL (rat, m) = 300 mg/kg bw/d and NOAEL (rat, f) = 1000 mg/kg bw/d. [Read-across data from tributyl-O-acetylcitrate (CAS 77-90-7)]
3) Finkelstein, 1959: Tributyl citrate did not induce deleterious effects at concentration of 5%, but the 10%-diet tended to depress the growth, an effect which may be due to frequent diarrhoea. Sections taken from the treated animals were indistinguishable from the controls.
4) Finkelstein, 1959: There were no symptoms of toxicity and the tests showed no abnormalities.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records
Reference
Endpoint:
sub-chronic toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Study period:
2001 - 2002
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: Comparable to Guideline study
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity in Rodents)
Deviations:
yes
Remarks:
Range-finding study: Only limited histopathology performed and no special neurotoxicity examination included
Principles of method if other than guideline:
Range-finding study: Only limited histopathology performed and no special neurotoxicity examination included
GLP compliance:
no
Remarks:
no GLP required (range-finding study for long-term toxicity)
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Route of administration:
oral: feed
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
yes
Duration of treatment / exposure:
13 weeks
Frequency of treatment:
Daily via the diet
Remarks:
Doses / Concentrations:
0, 96.02, 287.50 and 961.16 mg/kg
Basis:
actual ingested
No. of animals per sex per dose:
10 m / 10 f
Control animals:
yes, plain diet
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
Observation for mortality/viability were recorded twice daily

DETAILED CLINICAL OBSERVATIONS: Yes
Time schedule: At least once daily. Palpation for tissue masses were performed at least once weekly

BODY WEIGHT: Yes
Time schedule for examinations: Weekly during pre-test, treatment and before necropsy

FOOD CONSUMPTION AND COMPOUND INTAKE:
Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day

FOOD EFFICIENCY: no

WATER CONSUMPTION AND COMPOUND INTAKE: no

OPHTHALMOSCOPIC EXAMINATION: Yes
Time schedule for examinations: During acclimatization and during week 13
Dose groups that were examined: All groups during acclimatization and all animals of the control and high dose group during week 13

HAEMATOLOGY: Yes
Time schedule for collection of blood: week 13 weeks
Anaesthetic used for blood collection: yes (light isoflurane anaesthesia)
Animals fasted: Yes
How many animals: all (10 m / 10 f per group)
Parameters examined: Erythrocyte count, haemoglobin, haematocrit, MCV, RDW, MCH, MCHC, HDW, reticulocyte, reticulocyte maturity index, leukocyte count, differential leukocyte count, prothrombin time, partial thromboplastin time

CLINICAL CHEMISTRY: Yes
Time schedule for collection of blood: week 13
Animals fasted: Yes
How many animals: all (10 m / 10 f per group)
Parameters examined: Glucose, urea, creatinine, bilirubin (total), cholesterol, tryglicerides, phospholipides, ASAT, ALAT, LDH, CK, ALP, GGT, Sodium, potassium, chloride, calcium, inorganic phosphorus, protein (total), albumin, globulin, albumin/globulin ration

URINALYSIS: no

NEUROBEHAVIOURAL EXAMINATION: no
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, all organs, all groups
HISTOPATHOLOGY: Yes.
Statistics:
Dunnett-test (many to one t-test) based on a pooled variance estimate if variables can be assumed to follow a normal distribution
Steel-test (many-one rank test) was applied instead of the Dunnett-test when data were not assumed to follow a normal distribution
Fisher's exact test was applied for ophthalmoscopic data and macroscopic findings
Armitage/Cochran Trend Test was used for non-neoplastic lesions, if appropriate
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:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
effects observed, treatment-related
Urinalysis findings:
not examined
Behaviour (functional findings):
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Gross pathological findings:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Histopathological findings: neoplastic:
no effects observed
Details on results:
CLINICAL CHEMISTRY
In m and f at 1000 mg/kg bw/day, decreased bilirubin levels, decreased aspartate aminotransferase and lactate dehydrogenase activity, increased sodium level, decreased chloride and calcium levels, decreased globulin levels (which resulted in increased albumin/globulin ratio) were evident in m at 300 and 1000 mg/kg bw/d

ORGAN WEIGHTS
1000 mg/kg: increased liver weights in m and f

GROSS PATHOLOGY
1000 mg/kg: enlarged livers in 2 f

HISTOPATHOLOGY: NON-NEOPLASTIC
1000 mg/kg: minimal hepatocellular hypertrophy (non-adverse) in several animals
Dose descriptor:
NOAEL
Remarks:
1000 mg/kg bw/day
Effect level:
>= 1 000
Sex:
male/female
Basis for effect level:
other: see 'Remark'
Critical effects observed:
not specified

Table: Summary of test item-related findings for ATBC

 

Intended test item intake (mg/kg bw/day)

0

100

300

1000

 

 

 

 

 

Clinical chemistry

Males (means)

 

 

 

 

Glucose (mmol/L)

4.886

6.124**

5.895**

5.784*

Bilirubin (µmol/L)

2.055

1.955

1.767

1.305**

ASAT (U/L)

82.37

76.37

71.90**

66.08**

LDH (U/L)

136.50

120.88

92.29**

91.27**

Sodium (mmol/L)

144.58

145.00

154.52**

153.72**

Chloride (mmol/L)

100.94

101.33

89.18**

89.25**

Calcium (mmol/L)

2.834

2.811

2.666**

2.694**

Globulin (g/L)

26.356

26.474

24.431**

24.123**

Alb/Glob ratio (rel, %)

1.590

1.640

1.727*

1.755**

 

 

 

 

 

Females (means)

 

 

 

 

Bilirubin (µmol/L)

2.795

2.346

2.057

1.642**

 

 

 

 

 

Organ weights

Liver (means)

 

 

 

 

Males (abs., gram)

8.83

9.30

9.58

10.86**

Males (rel., %)

2.34

2.38

2.51

2.86**

 

 

 

 

 

Females (abs., gram

5.88

6.12

5.80

7.10**

Females (rel., %)

2.65

2.71

2.76

3.03*

 

 

 

 

 

Macroscopic findings

Liver (no. affected/10)

0/10

0/10

0/10

2/10

Females: Enlarged

 

 

 

 

Histopathology

Liver (no. affected/10)

 

 

 

 

Males: Hepatocellular hypertrophy

1/10

0/10

6/10

5/10

Females: Hepatocellular hypertrophy

0/10

0/10

1/10

2/10

 

  • * = 5% level (Anova and/or Dunnett-Test)
  • ** = 1% level (Anova and/or Dunnett-Test)

 

Conclusions:
Based on the results of this study, dose levels of 100, 300 and 1000 mg/kg bw/d were proposed for the subsequent 2-year combined chronic/carcinogenicity study. Slight effects seen at 1000 mg/kg bw/d were considered due to hepatic metabolism adaption. Thus the NOAEL for m and f in this study was given with 1000 mg/kg bw/d
Executive summary:

This 13-weeks dietary toxicity study with acetyl tributyl citrate (ATBC) in Wistar rats was designed as dose range finding study for a subsequent combined chronic/carcinogenicity study. In principle, the study was performed according to OECD Guideline 408, but with restricted histopathological organ examinations. The administration at target doses of 100, 300 and 1000 mg/kg bw/d resulted in no unscheduled deaths. No test item-related clinical signs were observed. Treatment-related findings were restricted to slight changes in clinical biochemistry (males were more affected than females) and slightly increased liver weights accompanied by minimal hepatocellular hypertrophy at 1000 mg/kg bw/d. The findings were considered to be due to hepatic metabolic adaption rather than as sign of toxicity. The highest dose of 1000 mg/kg bw can be regarded as NOAEL. The proposed dose levels for a subsequent combined chronic/carcinogenicity study were 100, 300 and 1000 mg/kg bw/d. It can be assumed that the same applies to tributyl citrate (CAS 77-94-1) as it is a near analogue to the test substance acetyl tributyl citrate.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
1 000 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
good quality

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: inhalation
Data waiving:
exposure considerations
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal
Data waiving:
other justification
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
Endpoint:
repeated dose toxicity: dermal
Data waiving:
other justification
Justification for data waiving:
other:
Critical effects observed:
not specified
Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Repeated dose toxicity: oral

A 13-week dietary toxicity study with acetyl tributyl citrate (ATBC) in Wistar rats was designed as a dose range finding study for a subsequent combined chronic/carcinogenicity study. In principle, the study was performed according to OECD Guideline 408, but with restricted histopathological organ examinations. The administration at target doses of 100, 300 and 1000 mg/kg bw/d resulted in no unscheduled deaths. No test item-related clinical signs were observed. Treatment-related findings were restricted to slight changes in clinical biochemistry (males were more affected than females) and slightly increased liver weights accompanied by minimal hepatocellular hypertrophy at 1000 mg/kg bw/d. The findings were considered to be due to hepatic metabolic adaption rather than as sign of toxicity. The highest dose of 1000 mg/kg bw can be regarded as the NOAEL. The proposed dose levels for a subsequent combined chronic/carcinogenicity study were 100, 300 and 1000 mg/kg bw/d. It can be assumed that the same applies to tributyl citrate (CAS 77-94-1) as it is a near analogue to the test substance acetyl tributyl citrate.

The repeated dose toxicity of tributyl citrate via the oral route was studied in 22 rats (Finkelstein & Gold, 1959). A diet representing a 5 % concentration of tributyl citrate had no deleterious effect on growth. The concentration of 10 % seemed to depress the growth curve. In view of the fact that the higher concentration tended to produce diarrhoea, it may well be that this factor accounts for the impairment in growth. A blood count was done before treatment, after feeding for 1 month, and again after feeding for 2 months. There are very wide fluctuations in the red cell count, white cell count, and differential count in both the control and treated animals. There is no conspicuous difference between the values in the controls and the treated animals. Eight rats were sacrificed after this period of feeding the diets containing 5 and 10 % tributyl citrate, and 2 control rats. The gross examination of the thoracic and abdominal viscera showed no abnormalities. Forty histological sections were made representing the following organs: heart, lungs, gastrointestinal tract, liver, pancreas, spleen, kidneys. The sections taken from the treated animals were indistinguishable from the controls. Further, in another test two cats were fed 5 mL/kg bw/day by stomach tube for 2 months. In addition to observations on the general behaviour and appearance of the animals, the following were done at approximately weekly intervals: weight, urine, blood NPW; blood creatinine; blood sugar and blood counts. Similar observations were made on the 2 untreated cats which were maintained under similar conditions during the same period. Each of the treated animals received approximately 50 doses of the compound.

There were no symptoms of toxicity and the tests showed no abnormalities. The treated animals developed loose bowel movements, and it is likely that the factor is responsible for the loss of approximately 30 % in body weight.


Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
Reliable subchronic study conducted with a structural analogue ATBC.

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
Inhalation is not a relevant route of exposure. An oral study is available, the oral NOAEL can be extrapolated to an inhalation NOAEC.

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
Inhalation is not a relevant route of exposure. An oral study is available, the oral NOAEL can be extrapolated to an inhalation NOAEC.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
Oral study is available. The oral NOAEL can be extrapolated to a dermal NOAEL.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
Oral study is available. The oral NOAEL can be extrapolated to a dermal NOAEL.

Justification for classification or non-classification

Repeated dose toxicity: oral The test material does not meet the criteria for classification and will not require labelling in accordance with European Regulation (EC) No. 1272/2008.