Registration Dossier

Administrative data

Description of key information

Oral: BASF AG, 1998. Subchronic oral toxicity study with 2-Pyrrolidone in Wistar rats; Administration in the drinking water for 3 months. Report No.52S0014/92038. GLP, according to the OECD guideline 408, rats, drinking water, 90 days, ca. 0, 37, 207, 586 and 1125 mg/kg bw/day.

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:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP Guideline study
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.26 (Sub-Chronic Oral Toxicity Test: Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Deviations:
no
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals or test system and environmental conditions:
Wistar rats (Chbb: THOM (SPF)) were obtained from Dr. Karl Thomae GmbH, Biberach/Riss, Germany
Route of administration:
oral: drinking water
Vehicle:
unchanged (no vehicle)
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Test solutions were analysed at the start and end of the study to assure that the concentrations were correct and the 4-day stability was assessed as 97 %. The mixtures were prepared at no less than 4-day intervals.
Duration of treatment / exposure:
3 months
Frequency of treatment:
continuously
Dose / conc.:
0 ppm
Remarks:
(ca. 0 mg/kg/d)
Dose / conc.:
600 ppm
Remarks:
(ca. 37 mg/kg/d)
Dose / conc.:
2 400 ppm
Remarks:
(ca. 207 mg/kg/d)
Dose / conc.:
7 200 ppm
Remarks:
(ca. 586 mg/kg/d)
Dose / conc.:
15 000 ppm
Remarks:
(ca. 1125 mg/kg/d)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
Post-exposure period: none
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: The animals’ state of health was checked each day.
- Cage side observations checked in table [No.?] were included.

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Animals were weighed weekly and given an additional comprehensive clinical examination / a thorough physical examination at each weighing.

BODY WEIGHT: Yes
- Time schedule for examinations: Animals were weighed weekly.
- The final bodyweight was recovered on day 91

FOOD CONSUMPTION: Yes
- Time schedule: Food consumption was determined weekly.

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: Yes / No / Not specified

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water consumption was determined once/week over a period of 4-days.

OPHTHALMOSCOPIC EXAMINATION: Yes / No / Not specified
- Time schedule for examinations:
- Dose groups that were examined:

HAEMATOLOGY: Yes
- Time schedule for collection of blood: blood was sampled and analyzed on study day 88
- Anaesthetic used for blood collection: Yes (identity) / No / Not specified
- Animals fasted: Yes / No / Not specified
- How many animals:
- Parameters examined: "complete haematology"

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: blood was sampled and analyzed on study day 88
- Animals fasted: Yes / No / Not specified
- How many animals:
- Parameters examined.: alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, serum-gammaglutamyltransferase, sodium, potassium, chloride, inorganic phosphate, calcium, urea, creatinine, glucose, total bilirubin, total protein, albumin, globulins, triglycerides, cholesterol, magnesium.

URINALYSIS: Yes
- Time schedule for collection of urine: Urine samples were taken on day 85
- Metabolism cages used for collection of urine: Yes / No / Not specified
- Animals fasted: Yes / No / Not specified
- Parameters examined.: "complete urinanalysis"

OTHER:
- necropsies were conducted over days 92 to day 95
Sacrifice and pathology:
At necropsy, major organs were weighed and sections were fixed for histopathology. All animals were subjected to gross-pathological assessment, followed by histopathological examination using a complete tissue list.
Statistics:
Means and standard deviations for the variables food consumption, body weight, body weight change, water consumption and test substance intake (except control group) were calculated for the animals of each test group. They were printed out in the summary and individual value tables, with the exception that for test substance intake and body weight change only summary tables were prepared. For the parameters food consumption, water consumption, body weight and body weight change a parametric one-way analysis of variance was done via the F-test (ANOVA). If the resulting p-values were equal to or less than 0.05, a comparison of each dose group with the control group was carried out. These comparisons were performed simultaneously via Dunnett’s test for the hypothesis of equal means. If the results of this test were significant, labels (* for p < 0.05, ** for p < 0.01) were printed together with the group means in the tables. Both tests were performed two-sided. Statistical analysis of histopathology was conduced with a proprietary computer program.
Clinical signs:
no effects observed
Description (incidence and severity):
No animal died during the study and no adverse clinical signs or ophthalmoscopic findings were noted.
Mortality:
no mortality observed
Description (incidence):
No animal died during the study and no adverse clinical signs or ophthalmoscopic findings were noted.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Body weight was impaired statistically significant in males and females of the 15000 ppm treatment and on few days also in females of the 7200 ppm treatment. On day 91, the values were 9 % (15000 ppm males and females), and 6 % (7200 ppm females) below controls, respectively. Body weight change was impaired statistically significant in males and females of the 7200 and 15000 ppm treatment. On day 91, the values were 14 % (15000 ppm males), 20 % (7200 ppm females), 7 % (7200 ppm males) and 16 % (7200 ppm males) below controls, respectively.
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
FOOD and WATER CONSUMPTION
Food consumption was impaired in the high dose treatment in males (-4 to -10 %) and females (-5 to -12 %) with statistical significance on several days. In addition, water consumption was significantly impaired in the high dose treatment in males (-5 to -19 %) and females (-14 to -27 %) with statistical significance on several days.
INTAKE OF TS
The approximate, mean daily intake of test substance in mg/kg body weight for the complete administration period is 33 -42 for the 600 ppm treatment, 184-230 for the 2400 ppm treatment, 529-643 for the 7200 ppm treatment and 1062-1189 for the 15000 ppm treatment.
Food efficiency:
not specified
Water consumption and compound intake (if drinking water study):
effects observed, treatment-related
Description (incidence and severity):
Water consumption was significantly impaired in the high dose treatment in males (-5 to -19 %) and females (-14 to -27 %) with statistical significance on several days.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No animal died during the study and no adverse clinical signs or ophthalmoscopic findings were noted.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
Hematology: No substance-related changes were observed in the results of the hematology examinations of both sexes.
Clotting analyses: At the end of the study statistically significant prolonged prothrombin times were measured in the plasma of the high dose males. In the high dose females there was only a trend towards prolonged prothrombin times.
Enzymes: No substance-related changes were observed in the results of the serum enzyme examinations of both sexes.
Clinical biochemistry findings:
effects observed, treatment-related
Description (incidence and severity):
Blood chemistry: After 3 months of test substance administration statistically significant decreased total protein and globulin concentrations were found in the serum of the male and female animals of the high dose treatment. Additionally, statistically significant decreased total protein levels were detected in the females of the 7200 ppm treatment. In the high dose males, triglyceride concentrations were statistically significant increased, whereas in the high dose females this finding was seen only as a trend. Moreover, statistically significant reduced creatinine levels were found in females of the two highest dosed treatments; in the males of the two highest dosed treatments only a trend towards decreased creatinine concentrations was observed.
Urinalysis findings:
effects observed, treatment-related
Description (incidence and severity):
Increased specific gravity and reduced volume were found in the urine specimens of the male animals of the two highest dosed treatments. Furthermore, in most of the urine specimens of the males of the two highest dosed treatments a dark-yellow discoloration was noted. In the females no corresponding findings were observed.
Behaviour (functional findings):
not specified
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Description (incidence and severity):
Relative kidney weights were increased in males and females of the 15000 ppm treatment and in males of the 7200 ppm treatment. In females of all treatments with the TS an altered cellular composition of the thymic cortex was seen.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
In females of all treatments with the TS an altered cellular composition of the thymic cortex was seen.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
no effects observed
Other effects:
not examined
Details on results:
Mean test material consumption in mg/kg*day were: - males: 33, 184, 529 and 1062 mg/kg - females 42, 230, 643 and 1189 mglkg

MORTALITY, CLINICAL SIGNS and OPHTHALMOSCOPY
No animal died during the study and no adverse clinical signs or ophthalmoscopic findings were noted.

FOOD and WATER CONSUMPTION
Food consumption was impaired in the high dose treatment in males (-4 to -10 %) and females (-5 to -12 %) with statistical significance on several days. In addition, water consumption was significantly impaired in the high dose treatment in males (-5 to -19 %) and females (-14 to -27 %) with statistical significance on several days.

BODY WEIGHT and BODY WEIGHT GAIN
Body weight was impaired statistically significant in males and females of the 15000 ppm treatment and on few days also in females of the 7200 ppm treatment. On day 91, the values were 9 % (15000 ppm males and females), and 6 % (7200 ppm females) below controls, respectively. Body weight change was impaired statistically significant in males and females of the 7200 and 15000 ppm treatment. On day 91, the values were 14 % (15000 ppm males), 20 % (7200 ppm females), 7 % (7200 ppm males) and 16 % (7200 ppm males) below controls, respectively.
INTAKE OF TS The approximate, mean daily intake of test substance in mg/kg body weight for the complete administration period is 33 -42 for the 600 ppm treatment, 184-230 for the 2400 ppm treatment, 529-643 for the 7200 ppm treatment and 1062-1189 for the 15000 ppm treatment.

CLINICAL CHEMISTRY, HEMATOLOGY and URINALYSES
Hematology: No substance-related changes were observed in the results of the hematology examinations of both sexes.
Clotting analyses: At the end of the study statistically significant prolonged prothrombin times were measured in the plasma of the high dose males. In the high dose females there was only a trend towards prolonged prothrombin times.
Enzymes: No substance-related changes were observed in the results of the serum enzyme examinations of both sexes.
Blood chemistry: After 3 months of test substance administration statistically significant decreased total protein and globulin concentrations were found in the serum of the male and female animals of the high dose treatment. Additionally, statistically significant decreased total protein levels were detected in the females of the 7200 ppm treatment. In the high dose males, triglyceride concentrations were statistically significant increased, whereas in the high dose females this finding was seen only as a trend. Moreover, statistically significant reduced creatinine levels were found in females of the two highest dosed treatments; in the males of the two highest dosed treatments only a trend towards decreased creatinine concentrations was observed.
Urinalyses: Increased specific gravity and reduced volume were found in the urine specimens of the male animals of the two highest dosed treatments. Furthermore, in most of the urine specimens of the males of the two highest dosed treatments a dark-yellow discoloration was noted. In the females no corresponding findings were observed.

OTHER DEVIATIONS
There are some further statistically significant inter-group differences in the results of the clinical pathology testing. These deviations are marginal, sporadic or inconsistent, when compared with the other sex, or lack dosage-relationship. Accordingly, these changes are considered to be of no toxicological significance.

PATHOLOGY
Relative kidney weights were increased in males and females of the 15000 ppm treatment and in males of the 7200 ppm treatment. In females of all treatments with the TS an altered cellular composition of the thymic cortex was seen.

CONCLUSION
The kidney appears to be a target organ at dose levels of 7200 ppm (about 586 mg/kg) in the drinking water and above. The NOAEL is 2400 ppm in drinking water or about 207 mg/kg*bw*day. In all treated females an altered cellular composition of the thymic cortex was seen. This was assessed as being incidental, due to the following reasons: 1. there was no dose-response relationship 2. only females were affected 3. in randomly selected control groups of other subchronic studies the same finding was observed in several cases, also.
Dose descriptor:
NOAEL
Effect level:
207 mg/kg bw/day (nominal)
Sex:
male/female
Basis for effect level:
other: Effects on kidneys
Critical effects observed:
not specified

Mean test material consumption in mg/kg*day were:
- males: 33, 184, 529 and 1062 mg/kg
- females 42, 230, 643 and 1189 mglkg

MORTALITY, CLINICAL SIGNS and OPHTHALMOSCOPY
No animal died during the study and no adverse clinical signs or ophthalmoscopic findings were noted.

FOOD and WATER CONSUMPTION
Food consumption was impaired in the high dose treatment in males (-4 to -10 %) and females (-5 to -12 %) with statistical significance on several days. In addition, water consumption was significantly impaired in the high dose treatment in males (-5 to -19 %) and females (-14 to -27 %) with statistical significance on several days.

BODY WEIGHT and BODY WEIGHT GAIN
Body weight was impaired statistically significant in males and females of the 15000 ppm treatment and on few days also in females of the 7200 ppm treatment. On day 91, the values were 9 % (15000 ppm males and females), and 6 % (7200 ppm females) below controls, respectively.
Body weight change was impaired statistically significant in males and females of the 7200 and 15000 ppm treatment. On day 91, the values were 14 % (15000 ppm males), 20 % (7200 ppm females), 7 % (7200 ppm males) and 16 % (7200 ppm males) below controls, respectively.

INTAKE OF TS
The approximate, mean daily intake of test substance in mg/kg body weight for the complete administration period is 33 -42 for the 600 ppm treatment, 184-230 for the 2400 ppm treatment, 529-643 for the 7200 ppm treatment and 1062-1189 for the 15000 ppm treatment.

CLINICAL CHEMISTRY, HEMATOLOGY and URINALYSES
Hematology: No substance-related changes were observed in the results of the hematology examinations of both sexes.
Clotting analyses: At the end of the study statistically significant prolonged prothrombin times were measured in the plasma of the high dose males. In the high dose females there was only a trend towards prolonged prothrombin times.
Enzymes: No substance-related changes were observed in the results of the serum enzyme examinations of both sexes.
Blood chemistry: After 3 months of test substance administration statistically significant decreased total protein and globulin concentrations were found in the serum of the male and female animals of the high dose treatment.
Additionally, statistically significant decreased total protein levels were detected in the females of the 7200 ppm treatment. In the high dose males, triglyceride concentrations were statistically significant increased, whereas in the high dose females this finding was seen only as a trend. Moreover, statistically significant reduced creatinine levels were found in females of the two highest dosed treatments; in the males of the two highest dosed treatments only a trend towards decreased creatinine concentrations was observed.
Urinalyses: Increased specific gravity and reduced volume were found in the urine specimens of the male animals of the two highest dosed treatments. Furthermore, in most of the urine specimens of the males of the two highest dosed treatments a dark-yellow discoloration was noted. In the females no corresponding findings were observed.

OTHER DEVIATIONS
There are some further statistically significant inter-group differences in the results of the clinical pathology testing. These deviations are marginal, sporadic or inconsistent, when compared with the other sex, or lack dosage-relationship. Accordingly, these changes are considered to be of no toxicological significance.

PATHOLOGY
Relative kidney weights were increased in males and females of the 15000 ppm treatment and in males of the 7200 ppm treatment. In females of all treatments with the TS an altered cellular composition of the thymic cortex was seen.

CONCLUSION
The kidney appears to be a target organ at dose levels of 7200 ppm (about 586 mg/kg) in the drinking water and above. The NOAEL is 2400 ppm in drinking water or about 207 mg/kg*bw*day.
In all treated females an altered cellular composition of the thymic cortex was seen. This was assessed as being incidental, due to the following reasons:
1. there was no dose-response relationship
2. only females were affected
3. in randomly selected control groups of other subchronic studies the same finding was observed in several cases, also.

Conclusions:
The test material is of low systemic toxicity if administered in repeated manner.
Executive summary:

2 -pyrrolidone was tested for its subchronic toxicity in a 3-month drinking water study. The study was conducted in compliance with the OECD Guideline 408 and the GLP.

The purpose of this study was an assessment of the systemic toxicity after repeated exposures to the test material. Wistar rats received 0, 37, 207, 586 and 1125 mg/kg/d 2-Pyrrolidone (corresponding to 600, 2400, 7200 and 15000 ppm) in the drinking water, respectively during 90 days.

All animals were subjected daily to clinical examinations. Food consumption, water consumption and body weight were determined each week. Urinalysis, clinicochemical and hematological examinations were carried out towards the end of the administration period. All animals were subjected to gross-pathological assessment, followed by histopathological examination.

No substance related effects were seen in the two lowest dose groups. In the mid-dose group all parameters checked were slightly adverse. In the high dose group, effects were dose-dependently more severe. The kidneys were the most affected organs.

Due to adverse effects on the renal system, the NOAEL is considered to be 207 mg/kg b.w. for male and female rats.

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
NOAEL
207 mg/kg bw/day
Study duration:
subchronic
Experimental exposure time per week (hours/week):
168
Species:
rat

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:oral

A 90 day sub-chronic study was conducted according to the GLP requirements with Wistar rats which received 0, 37, 207, 586 and 1125 mg/kg/d 2-Pyrrolidone (corresponding to 600, 2400, 7200 and 15000 ppm) in the drinking water, respectively (BASF AG, 1998). Food consumption, water consumption and body weight were determined each week. The animal’s state of health was checked each day. When the animals were weighed they were subjected to an additional comprehensive clinical examination. Ophthalmological examinations were carried out in the animals of the control group and highest dose group prior to the start and towards the end of the administration period. Urinalysis, clinicochemical and haematological examinations were carried out towards the end of the administration period. All animals were subjected to gross-pathological assessment, followed by histopathological examination.

No substance related effects were seen in the two lowest dose groups. In the 586 mg/kg treatment, slight decrease of food consumption in female animals, slight decrease of water consumption in both sexes, slightly decreased body weights in females, resulting in reduced values of 6 % an day 91, decreased body weight changes resulting in reduced values of 7 % (males) and 16 % (females) an day 91, decrease in creatinine in both sexes, decrease in total protein in the females, increased urinary specific gravity in the males, reduced urinary volume in the males, dark yellow discoloration of urine specimens in the males and increase in the mean relative kidney weights in males were observed. In the high dose group, effects were dose-dependently more severe (decreased food and water consumption in both sexes, decreased body weights resulting in reduced values of 9 % (males) and 8 % (females) on day 91, decreased body weight changes resulting in reduced values of 14 % (males) and 20 % (females) on day 91, prolonged prothrombin times in both sexes, decrease in total protein, globulins, triglycerides and creatinine in both sexes, increased urinary specific gravity in the males, reduced urinary volume in the males, dark yellow discoloration of urine specimens in the males and increase in the mean relative kidney weights in males and females).

Due to adverse effects on the renal system, the NOAEL is considered to be 207 mg/kg b.w. for male and female rats.

In this study, an altered cellular composition of the thymic cortex was seen in females of all treatment groups. This was assessed as being incidental, due to the following reasons: there was no dose-response relationship, only females were affected and in randomly selected control groups of other subchronic studies the same finding was observed in several cases, also.

Another GLP conform subchronic drinking water study was performed with only female Wistar rats to clarify this thymus findings (BASF AG 1998 b). Here, 5 females received 0, 5 and 1339 mg/kg b.w., respectively (corresponding to 0, 50, 15000 ppm) for 90 days, satellite groups of the control and the high dose group recovered for additional 28 days. Results indicate that the formerly described thymic alterations were not test substance-related. The oestrous cycle determinations and haematological examinations did not reveal a hormonal or immunotoxic potential of the test substance. Deviations in the weight parameters were regarded as incidental.

Justification for classification or non-classification

Classification is not warranted according to the criteria of EU Classification, Labelling and Packaging of Substances and Mixtures (CLP) Regulation (EC) No. 1272/2008.