γ-valerolactone

Endpoint summary

Description of key information

Repeated dose toxicity, oral

The toxicity of the test item after repeated dose exposure was tested in three studies. The administration of the test item by gavage to female Wistar rats for 2 weeks caused no treatment-related, relevant effects at a dose of 1000 mg/kg bw. Thus, the NOAEL was found to be ≥1000 mg/kg bw in this study. A 90-Day feeding study in rats receiving dietary doses of the test item revealed no evidence of adverse toxic effects (NOAEL ≥ 49 (males) or 51.1 (females) mg/kg bw). A 13 week feeding study in rats showed no treatment-related effects at doses of 500 mg/kg bw. Thus the NOAEL was found to be ≥500 mg/kg bw.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Reference 1

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not available, publication received 1966-11-07

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: The subacute and/or chronic toxicity of 48 food flavourings was studied in rats. A summary of the studies is presented.
- Short description of test conditions: 0, 1000, 2500, 5000 and 10000 ppm of the test item (corresponding to 0-500 mg/kg bw) were given to rats via. feed for 13 weeks.
- Parameters analysed / observed: Body weights were recorded weekly and hematological examinations were made at termination of the study. Organ weights of kidneys, spleen, heart, and testes were recorded. These organs, the remaining abdominal and thoracic viscera, and one hind leg, for bone, bone marrow and muscle, were preserved in 10 % buffered formalin-saline solution for histopathological examination.

GLP compliance:

not specified

Limit test:

no

Species:

rat

Strain:

other: Weanling Osborne-Mendel

Details on species / strain selection:

Not specified

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Housing: Individually in wire cages
- Diet: Ad libitum
- Water: Ad libitum

Route of administration:

oral: feed

Details on route of administration:

Not specified

Vehicle:

not specified

Details on oral exposure:

DIET PREPARATION
- Rate of preparation of diet: Fresh diets were made and distributed weekly

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Not specified.

Dose / conc.:

10 000 ppm

Remarks:

equivalent to 500 mg/kg bw based on the conversion factors for old rats available in the JECFA guidelines for the preparation of toxicological working papers on Food Additives (JECFA, 2000)

Dose / conc.:

5 000 ppm

Remarks:

equivalent to 250 mg/kg bw based on the conversion factors for old rats available in the JECFA guidelines for the preparation of toxicological working papers on Food Additives (JECFA, 2000)

Dose / conc.:

2 500 ppm

Remarks:

equivalent to 125 mg/kg bw based on the conversion factors for old rats available in the JECFA guidelines for the preparation of toxicological working papers on Food Additives (JECFA, 2000)

Dose / conc.:

1 000 ppm

Remarks:

equivalent to 50 mg/kg bw based on the conversion factors for old rats available in the JECFA guidelines for the preparation of toxicological working papers on Food Additives (JECFA, 2000)

No. of animals per sex per dose:

10000 ppm: 11 males + 10 females
2500 ppm: 11 males + 9 females
5000 ppm: Not indicated
1000 ppm: 10 males + 10 females

Control animals:

yes

Details on study design:

- Rationale for animal assignment: Litter mates were used except for a few subacute studies in which the weanlings were randomized by weight (every level having animals of equal weight).

Positive control:

Not specified

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Not specified

DETAILED CLINICAL OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: Weekly

FOOD CONSUMPTION AND COMPOUND INTAKE: Yes
- Food consumption was determined weekly, calculation method is not specified

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: Not specified

OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At the termination of the experiments the rats were sacrificed and exsanguinated.
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- How many animals: All animals
- Parameters checked in table 1 were examined.

CLINICAL CHEMISTRY: Not specified

PLASMA/SERUM HORMONES/LIPIDS: Not specified

URINALYSIS: Not specified

NEUROBEHAVIOURAL EXAMINATION: Not specified

IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes

HISTOPATHOLOGY: Yes (see "Any other information on material and methods incl. tables")

Optional endpoint(s):

Optional endpoints: No

Other examinations:

No other examinations were indicated

Statistics:

Not specified.

Clinical signs:

not specified

Mortality:

no mortality observed

Description (incidence):

Not specified

Body weight and weight changes:

no effects observed

Description (incidence and severity):

Not specified

Food consumption and compound intake (if feeding study):

no effects observed

Description (incidence and severity):

Not specified

Food efficiency:

not specified

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

not specified

Ophthalmological findings:

not examined

Haematological findings:

no effects observed

Description (incidence and severity):

Not specified

Clinical biochemistry findings:

not specified

Endocrine findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

no effects observed

Description (incidence and severity):

Not specified

Gross pathological findings:

no effects observed

Description (incidence and severity):

Not specified

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

no effects observed

Description (incidence and severity):

Not specified

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

>= 500 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male/female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Reference 2

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

weight of evidence

Study period:

Not available, publication received 1965-05-17

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

no guideline followed

Principles of method if other than guideline:

- Principle of test: Subchronic feeding study, not all parameters of a guideline study were investigated.
- Short description of test conditions: Dosages were administered on a uniform body weight basis by weekly adjustments of the concentration of the test material in the cotton-seed oil.
- Parameters analysed / observed: Body weight, clinical chemistry, haematology, histopathology

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

The substance supplied were typical of the commercial grade and were not chemically pure in all instances.

Species:

rat

Strain:

other: FDRL strain

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Housing: Individually in wiremesh cages
- Diet: Ad libitum
- Water: Ad libitum

Route of administration:

oral: feed

Details on route of administration:

Dosages were administered on a uniform body weight basis by weekly adjustments of the concentration of the test material in the cotton-seed oil. Food consumption records were maintained to compare the observed with the expected dosages of the test materials.

Vehicle:

cotton seed oil

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
Each test substance was diluted in cotton-seed oil in a concentration sufficient to provide the predetermined dosage in 2% of the diet.

DIET PREPARATION
- Mixing appropriate amounts with: Purina Laboratory Chow

VEHICLE
- Justification for use and choice of vehicle: Cotton seed oil is a widely used vehicle for feeding studies
- Concentration in vehicle: In a concentration sufficient to provide the predetermined dosage in 2% of the diet

Analytical verification of doses or concentrations:

not specified

Details on analytical verification of doses or concentrations:

Not specified

Duration of treatment / exposure:

90 days

Frequency of treatment:

Not indicated

Dose / conc.:

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

Remarks:

females

Dose / conc.:

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

Remarks:

males

No. of animals per sex per dose:

15 animals per sex per dose

Control animals:

yes

Details on study design:

- Dose selection rationale: Dosage levels were derived from the human total estimated daily intake, calculated on a mg/kg body weight basis assuming 50 kg as the average body weight, and multiplying by 100. Thus a "no-effect" response at this dosage would indicate that the estimated maximum use level complied with the application of a 1 :100 safety factor.

Positive control:

Not indicated

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Not specified

DETAILED CLINICAL OBSERVATIONS: Not specified

BODY WEIGHT: Yes
- Time schedule for examinations: Not specified

FOOD CONSUMPTION AND COMPOUND INTAKE:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Not specified
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: Not specified

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: Not specified


OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: Yes
- Time schedule for collection of blood: At a 6-week period (on 8 rats of each sex), at 12 weeks (in all rats)
- Anaesthetic used for blood collection: Not specified
- Animals fasted: Not specified
- Parameters examined: hematocrit (%), hemolglobin (g/100 mL), red blood cells (x 10^6/mm³), white blood cells (x 10^3/mm³), neutrophils (%), lymphocytes (%), blood urea nitrogen (mg/100 mL)

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: At a 6-week period (on 8 rats of each sex), at 12 weeks (in all rats)
- Animals fasted: Not specified

PLASMA/SERUM HORMONES/LIPIDS: Not specified

URINALYSIS: Not specified

NEUROBEHAVIOURAL EXAMINATION: Not specified

IMMUNOLOGY: Not specified

Sacrifice and pathology:

GROSS PATHOLOGY: Not specified

HISTOPATHOLOGY: Yes (see table 1)

Optional endpoint(s):

Optional endpoints: No

Other examinations:

No

Statistics:

Not specified

Clinical signs:

not specified

Mortality:

not specified

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

no effects observed

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

not specified

Ophthalmological findings:

not specified

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

Both sexes responded with reduced leucocyte counts, despite normal differential counts, and in the case of the males, slightly lower red cell counts than the corresponding controls. The degree of difference was small when compared with the individual rather than the composite control group (6~99 versus 8~08 x 10^6/mm³). None of the deviations shown for the test group can be regarded as
pathological even though they fall somewhat outside the statistical limits (P=0~05) for the controls .

Clinical biochemistry findings:

no effects observed

Endocrine findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

not specified

Immunological findings:

not specified

Organ weight findings including organ / body weight ratios:

no effects observed

Gross pathological findings:

no effects observed

Neuropathological findings:

not specified

Histopathological findings: non-neoplastic:

effects observed, non-treatment-related

Description (incidence and severity):

Various degrees of mild thyroid hyperplasia but this observation was also noted in the corresponding control groups and is not an uncommon finding in the laboratory. Despite these occasional aberrations, the opinion of the consultant pathologist who examined all slides resulting from this study was that none of the effects are be dose-related.

Histopathological findings: neoplastic:

not specified

Other effects:

not specified

Key result

Dose descriptor:

NOAEL

Effect level:

>= 51.1 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Dose descriptor:

NOAEL

Effect level:

>= 49 mg/kg bw/day (actual dose received)

Based on:

test mat.

Sex:

male

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Reference 3

Endpoint:

short-term repeated dose toxicity: oral

Remarks:

14-day repeated-dose oral toxicity study

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

2019-09-24 to 2019-11-06

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Justification for type of information:

A 14-day range-finding test with non-pregnant animals was conducted previous to an OECD 422 study with gamma-Valerolactone. The combined repeated dose toxicity study with the reprod./develop. tox. screening test, OECD 422, rats (gavage) is ongoing, the results will be submitted as soon as the data are available.

Qualifier:

equivalent or similar to guideline

Guideline:

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

Deviations:

yes

Remarks:

Not all parameters indicated in the guideline were investigated and a shorter period of time (14 days) was chosen.

Principles of method if other than guideline:

- Principle of test: 14-day range-finding test study to obtain initial information on the effect of the test substance after repeated oral administration (gavage) to non-pregnant, female Wistar rats before the beginning of subsequent maternal and prenatal toxicity studies.
- Short description of test conditions: Not all parameters indicated in the guideline were investigated.

GLP compliance:

yes

Limit test:

no

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Batch number of test material: Zimmerma 00003
- Expiration date of the batch: 12 Aug 2021

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature

Species:

rat

Strain:

Wistar

Details on species / strain selection:

The rat is the preferred animal species for repeated dose toxicity studies according to the various test guidelines. This Wistar rat strain (Crl:Wl(Han)) is selected because extensive historical control data is available for these rats.

Sex:

female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services, Germany GmbH
- Females non-pregnant: Yes
- Age at study initiation: About 15 weeks
- Housing: Polysulfonate cages Typ 2000P (H-Temp) with dust-free wooden bedding. Cages were enriched with wooden gnawing blocks and large play tunnels.
- Diet: Ad libitum
- Water: Ad libitum
- Acclimation period: 29 days

DETAILS OF FOOD AND WATER QUALITY:
The food used in the study was assayed for chemical and microbial contaminants. Fed. Reg. Vol. 44, No. 91 (09 May 1979), p 27354 (EPA), served as the guideline for maximum tolerable contaminants. Additionally, the levels of phytoestrogens should not exceed 350 μg of genistein equivalents/gram. According to recommendations of the GVSOLAS, the total amount of bacteria must not exceed 1*10^5 per g food.

The drinking water is regularly assayed for chemical contaminants both by the municipal authorities of Frankenthal and by the Environmental Analytics Water/Steam Monitoring Department of BASF SE as well as for bacteria by a contract laboratory. The Drinking Water Regulation will serve as the guideline for maximum tolerable contaminants.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 45-65
- Air changes (per hr): 15
- Photoperiod (hrs dark / hrs light): 12/12

IN-LIFE DATES: From day 0 to day 14

Route of administration:

oral: gavage

Details on route of administration:

The oral administration of a test substance has been proven useful worldwide in numerous studies for discovering a potential toxicological profile.

Vehicle:

water

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:
For the test substance preparation, the specific amount of test substance is weighed, topped up with deionized water in a graduated flask and intensely mixed with a magnetic stirrer until it is completely dissolved. Before and during administration, the preparations is kept homogeneous with a magnetic stirrer.

VEHICLE
- Concentration in vehicle: 3 g/100 mL (for 300 mg/kg bw group) and 10 g/100 mL (for 1000 mg/kg bw group)
- Amount of vehicle: 10 mL/kg bw

Analytical verification of doses or concentrations:

no

Duration of treatment / exposure:

From day 0 to day 14

Frequency of treatment:

Daily

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Dose / conc.:

300 mg/kg bw/day (nominal)

Dose / conc.:

0 mg/kg bw/day (nominal)

No. of animals per sex per dose:

3 animals per sex per dose

Control animals:

yes

Details on study design:

- Dose selection rationale: Low and high dose levels as requested by the sponsor
- Rationale for animal assignment: According to their weight

Positive control:

No positive control included

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twice daily (mondays-fridays) or once daily (saturdays, sundays and public holidays)


DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Within 2 hours and between 2 and 5 hours after the administration, afterwards daily observations

BODY WEIGHT: Yes
- Time schedule for examinations: Body weights are recorded on days 0, 3, 7, 10, and 13.


OPHTHALMOSCOPIC EXAMINATION: No

HAEMATOLOGY: No

CLINICAL CHEMISTRY: No

PLASMA/SERUM HORMONES/LIPIDS: No


URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: No

IMMUNOLOGY: No

Sacrifice and pathology:

One day after the last administration, the animals were anesthetized with isoflurane, sacrificed by decapitation and assessed by gross pathology.

Optional endpoint(s):

Optional endpoints: No

Statistics:

Body weight and body weight change: DUNNETT-test (two-sided)

Clinical signs:

effects observed, non-treatment-related

Description (incidence and severity):

Females of the 300 and 1000 mg/kg bw group, effects not further specified

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 specified

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

no effects observed

Ophthalmological findings:

not examined

Haematological findings:

not examined

Clinical biochemistry findings:

not examined

Endocrine findings:

not examined

Urinalysis findings:

not examined

Behaviour (functional findings):

not examined

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

not examined

Gross pathological findings:

not examined

Neuropathological findings:

not examined

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Other effects:

not examined

Key result

Dose descriptor:

NOAEL

Effect level:

>= 1 000 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Remarks on result:

not determinable due to absence of adverse toxic effects

Key result

Critical effects observed:

no

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

500 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

Comparable to guideline study with acceptale restrictions

Repeated dose toxicity: inhalation - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated-dose toxicity (14 days), rat, RL2

A 14-day range-finding toxicity study has been conducted to obtain initial information on the effect of the test substance after repeated oral administration (gavage) to non-pregnant, female Wistar rats before the beginning of subsequent maternal and prenatal toxicity studies. Doses of 0, 300 and 1000 mg/kg bw in deionized water were given to three animals per dose. The test substance was administered to the animals orally by gavage for a maximum of 14 days. For this purpose, disposable syringes of suitable size and suitable gavage tubes were used. During the study, all animals were observed for any clinically abnormal signs. Furthermore, food and water consumption and body weights were recorded. One day after the last administration the animals were anesthetized with isoflurane, sacrificed by decapitation and assessed by gross pathology. The administration of the test item by gavage to female Wistar rats for 2 weeks caused no treatment-related, relevant effects. Thus, the NOAEL was found to be >1000 mg/kg bw (BASF 2019).

A Combined Repeated Dose Tox. Study with the Reprod./Develop. Tox. Screen. Test, OECD 422, rats (gavage) is ongoing, the results will be submitted as soon as the data are available.

 

Repeated-dose toxicity (90 days), rat, RL2

A 90-day repeated dose feeding study with the test item has been conducted in rats. Singe doses of 45.9 mg/kg bw in cotton seed oil were mixed with appropriate amounts of feed and given to 15 male and 15 female rats. The actual doses received were calculated as 49 mg/kg bw (males) and 51.1 mg/kg bw (females). Body weights, feed consumption, hematology, clinical chemistry and organ weights were evaluated. Based on the absence of treatment-related effects, the NOAEL was found to be >49 mg/kg bw (Oser et al., 1965).

 

Repeated-dose toxicity (13 weeks), rat, RL2

In the course of a screening of 48 food flavourings for repeated dose effects, 1000, 2500, 5000 and 10000 ppm of the test item (corresponding to 50-500 mg/kg bw) were given to rats via. feed for 13 weeks. Body weights were recorded weekly and hematological examinations were made at termination of the study. Organ weights of kidneys, spleen, heart, and testes were recorded. These organs, the remaining abdominal and thoracic viscera, and one hind leg, for bone, bone marrow and muscle, were preserved in 10 % buffered formalin-saline solution for histopathological examination. For routine histopathology, sections were embedded in paraffin wax and stained with haematoxylin and eosin. Tissues from rats dying during the experiment were examined for gross changes and were preserved if autolysis was not advanced. Organs were not weighed but abnormalities and the suspected reason for death were noted. No effects were found after treatment with the test item, thus the NOAEL was found to be >500 mg/kg bw (Hagan et al., 1966).

 

Repeated-dose toxicity, rat, RL4

Several possible metabolites of inethyl-n-butyl ketone were administered separately to rats in drinking water (0.25 - 1.0%, equals to 250-1000 mg/kg bw, as per the conversion factors for rat mentioned in the SOP) or by gavage (0.2-1.2 mg/kg). No effects were found, thus the NOAEL was found to be >1000 mg/kg bw. No further details were given (Opdyke 1962-1980, original study by Krasavage, O'Donoghue & Terhaar, 1978)

 

 

Justification for classification or non-classification

Classification, Labeling, and Packaging Regulation (EC) No 1272/2008

Using a weight of evidence approach, the repeated dose toxicity of the test item was assessed. The available experimental test data are reliable and suitable for classification purposes under Regulation (EC) No 1272/2008. The test substance showed no toxicity after repeated exposure to rats, thus it is not considered to be classified for repeated dose toxicity under Regulation (EC) No 1272/2008, as amended for fifteenth time in Regulation (EU) No 2020/217.