Registration Dossier

Administrative data

Description of key information

In the combined repeated dose toxicity study with the reproduction / developmental toxicity screening test (OECD 422) no adverse effects were observed for repeated dose toxicity up to the highest tested dose (750 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:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2018-07-17 to 2018-09-18
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
GLP compliance:
yes (incl. certificate)
Limit test:
no
Species:
rat
Strain:
Wistar
Details on species / strain selection:
Wistar Han™:RccHan™:WIST strain rats
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Envigo RMS (UK) Limited, Blackthorn, Bicester, Oxon, UK
- Females (if applicable) nulliparous and non-pregnant: [yes/no]
- Age at study initiation: males: approximately eleven weeks old and females: approximately twelve weeks old.
- Weight at study initiation: males: 283 to 367g and females: 190 to 232g
- Fasting period before study:
- Housing: in groups of three in solid floor polypropylene cages with stainless steel mesh lids and softwood flake bedding
- Diet (e.g. ad libitum): free access
- Water (e.g. ad libitum): free access
- Acclimation period: twenty days

DETAILS OF FOOD AND WATER QUALITY: A pelleted diet (Rodent 2018C Teklad Global Certified Diet, Envigo RMS (UK) Limited, Oxon, UK.) was used. Certificates of analysis of the batches of diet used are given in the original report. Mains drinking water was supplied from polycarbonate bottles attached to the cage.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3 °C
- Humidity (%): 50 ± 20%
- Air changes (per hr): at least fifteen air changes per hour
- Photoperiod (hrs dark / hrs light): 12 hrs dark / 12 hrs light

IN-LIFE DATES: From: To: The in-life phase of the study was conducted between 17 July 2018 (first day of treatment) and 18 September 2018 (final day of necropsy).

Humidity:
The Study Plan target value for relative humidity (RH) was 50 ± 20%. This target range was exceeded on several occasions during the study with a maximum relative humidity of 74% being reached. Whilst these deviations from the target range were regrettable, the condition of the animals was closely monitored and there was no indication that the incidence of high humidity had any impact on the health of the animals. It was therefore considered that the purpose and integrity of the study had been unaffected.

Route of administration:
oral: gavage
Vehicle:
other: Dried Propylene Glycol
Details on oral exposure:
The test item was administered daily by gavage using a rubber dosing catheter attached to a graduated disposable plastic syringe. Control animals were treated in an identical manner with 4 mL/kg of Dried Propylene Glycol.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Samples of test item formulations were taken on two occasions and analyzed for concentration of the substance. The results indicate that the prepared formulations were within 95-100% of the nominal concentration indicating acceptable accuracy of the formulation procedure.
Duration of treatment / exposure:
Four weeks for males and at least seven weeks for females (including a two week pre-pairing phase, pairing, gestation and lactation phases)
Frequency of treatment:
once/day
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
750 mg/kg bw/day (nominal)
No. of animals per sex per dose:
12
Control animals:
yes, concurrent vehicle
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Not specified

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: immediately before dosing, soon after dosing, and one hour after dosing during the working week (except for females during parturition where applicable).

Morbidity/Mortality inspections were scheduled in the Study Plan to take place, twice daily, early and late during the working period. On three occasions (twice during the acclimatization period and on the penultimate day of the in-life phase of the study), the morning check of the animals was performed later than normally considered acceptable. Whilst this is regrettable, there was no indication that this resulted in any delay in detecting effects on the animal’s health during the study and as such, there was no impact on the scientific integrity of the study.

Post-dosing observations were not recorded on Day 26 of the study in error. Based on the post-dosing observation observed both prior to and after this occurrence, it is considered that no significant clinical signs would have been missed, due to this oversight. This deviation is, therefore, considered to have had no impact on the scientific integrity of the study.

BODY WEIGHT: Yes
- Time schedule for examinations: On Day 1 (prior to dosing) and then weekly for males until termination and weekly for females until pairing. During pairing phase females were weighed daily until mating was confirmed. Body weights were then recorded for females on Days 0, 7, 14 and 20 post coitum, and on Days 1, 4, 7 and 13 post partum. Body weights were also recorded at terminal kill.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
During the pre-pairing period, weekly food consumption was recorded for each cage of adults. For females showing evidence of mating, food consumption was recorded for the periods covering post coitum Days 0-7, 7-14 and 14-20. For females with live litters, food consumption was recorded for the periods covering post partum Days 1-4, 4-7 and 7-13.

FOOD EFFICIENCY:
%Food Efficiency = (Group mean bodyweight gain (g/rat/period) / Group mean food consumption (g/rat/day) x number of days) x 100
Food efficiency (the ratio of body weight change/dietary intake) was calculated retrospectively for males and females during the pre-pairing phase. Due to offspring growth and milk production, food efficiency could not be accurately calculated during gestation and lactation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: Water intake was measured gravimetrically on a daily basis from Day 2 of the study throughout the pre-pairing period.

OPHTHALMOSCOPIC EXAMINATION: Not specified

HAEMATOLOGY: Yes
Hematological and blood chemical investigations were performed on five males and five females selected from each test and control group prior to termination (Day 28 for males and Day 13 post partum for females). Blood samples were obtained from the lateral tail vein. Where necessary repeat samples were taken by cardiac puncture at termination. Animals were not fasted prior to sampling.
- Parameters checked in table [No.4] were examined.

CLINICAL CHEMISTRY: Yes
Hematological and blood chemical investigations were performed on five males and five females selected from each test and control group prior to termination (Day 28 for males and Day 13 post partum for females). Blood samples were obtained from the lateral tail vein. Where necessary repeat samples were taken by cardiac puncture at termination. Animals were not fasted prior to sampling.
- Parameters checked in table [No.5] were examined.

URINALYSIS: No


BEHAVIORAL ASSESSMENT: Yes
- Time schedule for examinations: Prior to the start of treatment and at approximately weekly intervals thereafter
- Dose groups that were examined: all
- Parameters checked in table [No.7] were examined.


IMMUNOLOGY: No


OTHER:
Functional observations:
Prior to the start of treatment and at approximately weekly intervals thereafter, all animals were observed for signs of functional/behavioral toxicity. These observations were performed on mated females on Days 4, 11 and 18 post coitum and for littering females on Days 4 and 12 post partum. Functional performance tests were also performed on five selected males and females from each dose level, prior to termination, together with an assessment of sensory reactivity to various stimuli.
No functional observations were performed for Females 38 and 43 on Day 18 of gestation in error. Although this loss of data is regrettable, there is considered to be sufficient data from the remaining animals to draw meaningful conclusions from. The purpose and scientific integrity of the study is therefore unaffected.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
Adult males were killed by intravenous overdose of suitable barbiturate agent followed by exsanguination on Day 29. Adult females were killed by intravenous overdose of suitable barbiturate agent followed by exsanguination on Day 12 post partum. All adult animals, including those dying during the study, were subjected to a full external and internal examination, and any macroscopic abnormalities were recorded.

HISTOPATHOLOGY: Yes (see table 6)
Samples of the following tissues were removed from five selected males and five selected females from each dose group and preserved in buffered 10% formalin, except where stated.
Statistics:
Data on statistics is stated in section "Any other information on materials and methods incl. tables"
Clinical signs:
no effects observed
Description (incidence and severity):
Instances of increased post-dosing salivation were observed for the majority of animals (10/12 males and 11/12 females) receiving 750 mg/kg bw/day.

Instances of increased post-dosing salivation were observed for the majority of animals (10/12 males and 11/12 females) receiving 750 mg/kg bw/day. The incidence of this clinical sign at this dosage appeared to be slightly higher for females than for males , but this may have reflected the comparatively longer treatment period for females compared to their male counterparts. Increased post-dosing salivation was also observed for the majority of animals (7/12 males and 8/12 females) receiving 300 mg/kg bw/day, although the incidence of this sign, which was similar in both sexes, appeared lower than that observed at 750 mg/kg bw/day. At 100 mg/kg bw/day, increased post-dosing salivation was restricted to just two females, each on only a single occasion towards the end of their treatment period. Increased post-dosing salivation is often observed when animals are dosed via the oral gavage route and are generally considered to reflect slight distaste or irritancy of the test item rather than a systemic effect of treatment.
Occasional instances of noisy respiration were observed for two females at 300 mg/kg bw/day and three males and four females at 750 mg/kg bw/day during the study, with one of the females at 750 mg/kg bw/day also showing noisy respiration during the assessment of behavioral observations. Additionally, one female at 100 mg/kg bw/day and one male at 750 mg/kg bw/day showed noisy respiration during the assessment of behavioral observations. Although the incidence of noisy respiration was higher at the high dosage of 750 mg/kg bw/day, the instances of noisy respiration tended to be quite isolated and, overall, the occurrence of this clinical sign was considered to reflect occasional difficulties in dosing isolated animals and to be of no toxicological significance .
One female treated with 300 mg/kg bw/day showed generalized fur loss from Day 24 of the study, but, in isolation, this finding was considered to be incidental and unrelated to treatment.
Summary incidence of daily clinical obserbations are given in table 1.


Mortality:
no mortality observed
Description (incidence):
There were no unscheduled deaths during the study.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
Group mean weekly body weights and standard deviations are given in Table 2 and 3. Group mean weekly body weight gains and standard deviations are given in Table 4 to 6.
Males
At 750 mg/kg bw/day body weight gain of males tended to be lower than control throughout much of the treatment period, with differences attaining statistical significance during Weeks 2 and 4, and leading to statistically significantly lower overall body weight gain at termination on Day 29 of the study. There was no effect of treatment on body weight gain for males throughout the study at dosages of 100 or 300 mg/kg bw/day.
Females
There was no effect of treatment on body weight and body weight gain for females during the pre-pairing phase of the study at 100, 300 or 750 mg/kg bw/day.
At 750 mg/kg bw/day, mean body weight gains during the last week of gestation was slightly lower than control, leading to lower overall body weight gain, although differences from control failed to attain statistical significance. During the last week of treatment, body weight gain of the pregnant females is increasingly influenced by the growing litter and the lower gain observed at this dosage probably reflect lower litter weight/contribution from the gravid uterus rather than any underlying effect on maternal body weight. Supporting this, mean body weight on Day 1 of lactation was similar to control.
At 750 mg/kg bw/day, mean body weight gain of females tended to be lower than control throughout lactation, with cumulative body weight gain being statistically significantly lower than control on Days 7 and 14 of lactation.
Body weight gains during pregnancy and lactation was unaffected by treatment at 100 or 300 mg/kg bw/day.



Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
Group mean food consumptions are given in Table 7 and 8.

Males
There was no effect of treatment on food consumption of males during the pre-pairing phase of the study at 100, 300 or 750 mg/kg bw/day.
Females
There was no effect of treatment on food consumption of females during the pre-pairing phase of the study at 100, 300 or 750 mg/kg bw/day.
At 750 mg/kg bw/day, mean food consumption tended to be lower than control throughout gestation, although statistical significance was only apparent during the first week of pregnancy. Subsequent mean food consumption throughout lactation was statistically significantly lower than control at this dosage.
At 100 and 300 mg/kg bw/day, mean food consumption during pregnancy and lactation was essentially similar to control and unaffected by treatment at either dosage.


Food efficiency:
no effects observed
Description (incidence and severity):
Food efficiency for males and for females during the pre-mating phase is given in Table 9.

Males
At 750 mg/kg bw/day, food conversion efficiency was slightly lower than control during the second week of treatment, leading to slightly lower overall food conversion efficiency for the two week pre-pairing period, compared to control.
There was no effect of treatment on food conversion efficiency of males during the pre-pairing phase of the study at 100 or 300 mg/kg bw/day.
Females
There was no effect of treatment on food conversion efficiency of females during the pre-pairing phase of the study at 100, 300 or 750 mg/kg bw/day.
Water consumption and compound intake (if drinking water study):
no effects observed
Description (incidence and severity):
Water consumption of either sex was considered to be unaffected by treatment at 100, 300 or 750 mg/kg bw/day.

There was no clear effect of treatment on water consumption during the pre-pairing treatment phase for either sex at 100 300 or 750 g/kg bw/day.
For females at 750 mg/kg bw/day water consumption appeared higher than control during the last five days of treatment but there was considered to be insufficient consistency between the individual cages at this dosage to indicate a treatment- related effect.
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
Intergroup differences for hematology parameters at the end of the treatment period did not indicate any effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.
For females at 750 mg/kg bw/day, mean reticulocytes counts were statistically significantly higher than control, with 4/5 individual values exceeding the historical control. In the absence of any supporting statistically significant differences from control for other erythrocyte parameters or any supporting histopathological change, this isolated finding was considered to be incidental and unrelated to treatment.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
There was no effect of treatment on blood chemistry parameters for either sex at 100, 300 or 750 mg/kg bw/day.

Intergroup differences for blood chemistry parameters at the end of the treatment period did not indicate any adverse effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.
For males at 300 and 750 mg/kg bw/day, mean bile acid levels were statistically significantly higher than control, with 2/5 individual values at 300 mg/kg bw/day and 3/5 individual values at 750 mg/kg bw/day exceeding the historical control group. In the absence of any supporting histopathological change, this isolated finding was considered to be of no toxicological significance.
For males at 750 mg/kg bw/day, mean alanine aminotransferase activity was statistically significantly higher than control, but only 1/5 individual values for these treated animals were within the historical control range. In the absence of any supporting histopathological change, this isolated finding was considered to be incidental and unrelated to treatment.
For females at 100 and 300 mg/kg bw/day, mean albumin/globulin (A/G) ratio was statistically significantly lower than control, however, there was no corresponding statistically significant differences apparent for mean levels of total protein or albumin at these dosages. All individual values for the A/G ratio of these treated females were within the historical control range and, in the absence of any similar effect at 750 mg/kg bw/day or supporting histopathological change, this finding was considered to be incidental and unrelated to treatment.
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
- Behavioral Assessment
Behavioral assessment throughout the study did not indicate any effect of treatment at 100, 300 or 750 mg/kg bw/day.
At 100 mg/kg bw/day, noisy respiration was observed for one female during the second week of lactation. At 750 mg/kg bw/day, noisy respiration was observed for one male during Week 5 and one female during the first week of lactation. These isolated instances of noisy respiration were consistent with similar instances observed during routine assessments of clinical condition and were considered to reflect occasional difficulties in dosing isolated animals. As such, they were considered not to indicate any underlying neurological effect on the behavior of the animals.
- Functional performance
Functional performance tests did not indicate any obvious effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.
At 100 mg/kg bw/day, overall motor activity for males was statistically significantly higher than control, however in the absence of any similar effect at higher dosages, this finding was considered incidental and unrelated to treatment.
- Sensory reactivity assessments
Sensory reactivity assessments did not indicate any effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, non-treatment-related
Description (incidence and severity):
Intergroup differences for organ weights did not indicate any adverse effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.

For males at 750 mg/kg bw/day, mean absolute and body weight relative testes weights were statistically significantly higher than control. For these treated males, 1/12 individual absolute weights and 4/12 individual relative weights exceeded the respective historical control range however, one absolute value was below the historical control range. For the control group 2/12 individual absolute weights were below and 1/12 individual relative weights exceeded the historical control range. In the absence of any effect on fertility on the study or evidence of histopathological change at 750 mg/kg bw/day, this finding was considered likely to be incidental and to be of little toxicological significance.
For females at 300 mg/kg bw/day , mean absolute and body weight relative liver weights were statistically significantly higher than control. For this organ, body weight relative values are normally regarded as being the best indicator of toxicological effect and all individual body weight relative values for these treated females were within the historical control range, while one control value exceeded this historical range. In the absence of any similar increase in liver weights or supporting histopathological liver changes for females at 750 mg/kg bw/day, this finding at 300 mg/kg bw/day was considered to be incidental and unrelated to treatment.
For males at 100 mg/kg bw/day, mean absolute and body weight relative prostate weights were statistically significantly higher than control. However, all individual absolute and body weight relative values were within the respective historical control range. In the absence of similar findings at higher dosages, this finding was considered incidental and unrelated to treatment.

Gross pathological findings:
no effects observed
Description (incidence and severity):
Macroscopic necropsy findings for adults did not indicate any effect of treatment for either sex at 100, 300 or 750 mg/kg bw/day.

Adults
The incidence, type and distribution of macroscopic findings observed at terminal necropsy of surviving animals did not indicate any effect of treatment at dosages of 100, 300 or 750 mg/kg bw/day.
One male treated with 750 mg/kg bw/day had small seminal vesicles, but this is considered unrelated to test item administration.
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
Microscopic examination of tissues did not indicate any effect of treatment at 750 mg/kg bw/day.

Adrenal Gland
Hypertrophy of the zona glomerulosa was present at a minimal level in 2/5 females treated at 750 mg/kg bw/day. Due to the low severity and the fact that the animals were lactating it is considered that this change is likely to be due to individual variation. Change in the zona glomerulosa is likely to be an adaptive response. The zona glomerulosa is the site of synthesis of aldosterone a mineralocorticoid which is mainly involved in the control of salt and water balance in the body.
Histopathological findings: neoplastic:
no effects observed
Description (incidence and severity):
Microscopic examination of tissues did not indicate any effect of treatment at 750 mg/kg bw/day.
Other effects:
effects observed, non-treatment-related
Description (incidence and severity):
Group mean values and standard deviations for for T4 and T3 in test and control group animals are given in Table 10 and Table 11, respectively.
At 750 mg/kg bw/day, mean thyroxine (T4) levels for adult males were lower than control, however, in the absence of any effect on organ weight or supporting histopathological change, this isolated finding was considered to be incidental and of no toxicological significance. No statistically significant differences from control were apparent for triiodothyronine (T3) levels for these adult males or for T4 or T3 levels in Day 13 offspring of either sex at this dosage.
Evaluation of T4 and T3 in adult males and offspring at Day 13 of age did not identify any obvious effect of treatment or indication of endocrine disruption at 100 or 300 mg/kg bw/day.




Details on results:
Treatment of males at 750 mg/kg bw/day was associated with lower body weight gains throughout the four week treatment period, although there was no effect on food consumption during the two week treatment pre-pairing period. For females at this dosage, body weight and food consumption during the two week pre-treatment were unaffected by treatment. Food consumption during gestation tended to be lower than control but there was no apparent effect on body weight gain. Lower body weight gain during the last week of gestation was observed but, this was considered to reflect a lower contribution from the gravid uterus due to a slightly lower litter weight rather than an underlying effect on maternal body weight, as body weight on Day 1 of lactation was similar to control. However, body weight gain and food consumption were lower than control throughout lactation and this occurred despite a slightly lower litter size than control at this dosage.
Clinical signs observed for the adult animals at 750 mg/kg bw/day did not indicate any systemic effect of treatment for either sex being confined to instances of increased post-dosing salivation and noisy respiration. There was also no effect of treatment on functional performance or behavioral observations.
Assessment of hematology and blood chemistry at 750 mg/kg bw/day did not indicate any adverse effect of treatment for either sex. For males, statistically significantly higher mean bile acid levels were apparent with 3/5 individual values exceeding the historical control group. However, the isolated nature of this finding and absence of supporting histopathological change, indicated that this finding was of no toxicological significance and did not represent an adverse effect. For females, mean reticulocytes counts were statistically significantly higher than control, with 4/5 individual values exceeding the historical control. There was no statistically significant differences from control for other erythrocyte parameters or supporting histopathological change, therefore this finding was considered to be incidental and, as such, not to represent an adverse effect.
Necropsy findings for adults animals at 750 mg/kg bw/day were unremarkable and did not indicate any effect of treatment. Statistically significant differences from control for organ weights were restricted to higher mean testes weights, but the majority of individual values for these treated males were within the historical control range. As there was no effect on fertility within the study or any histopathological change for this organ, this finding was considered likely to be incidental and not to represent an adverse effect. Microscopic evaluation of tissues at this dosage revealed hypertrophy of the adrenal zona glomerulosa at a minimal level for females, however, the low severity of this finding in lactating animals suggests that this change probably reflected normal individual variation.
At 750 mg/kg bw/day, mean thyroxine (T4) levels for adult males were statistically significantly lower than control, but there were no statistically significant differences from control apparent for triiodothyronine (T3) levels. The lower T4 values for males occurred in the absence of any effect on organ weight or supporting histopathological change and its isolated nature indicated that this finding was incidental and most likely unrelated to treatment.
At 100 and 300 mg/kg bw/day, clinical signs, functional performance or behavioral observations, body weight gain and food consumption did not indicate any systemic effect of treatment. For males at 300 mg/kg bw/day, mean bile acid levels were statistically significantly higher than control, but only 2/5 individual values exceeded the historical control group and in the absence of any supporting histopathological change at 750 mg//kg bw/day, this isolated finding appeared incidental and unrelated to treatment. Necropsy findings and organ weights did not indicate any effect of treatment at either dosage.


Dose descriptor:
NOEL
Effect level:
300 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
body weight and weight gain
other: food consumption
Key result
Dose descriptor:
NOAEL
Effect level:
750 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: absence of clear adverse effect of treatment
Key result
Critical effects observed:
no

Table 1 Summary Incidence of Daily Clinical Observations

                                                Day Numbers Relative to Start Date

Sex: Male

 

                                          Group 1               Group 2         Group 3         Group 4

                                        0(Control)         100 mg/kg bw/day 300 mg/kg bw/day 750 mg/kg bw/day

                                                                   

 

Scheduled kill                                                                                               

       Number of Animals                   12                    12              12              12     

       Days from ƒ to                    29 29                29 29          29 29          29 29   

                                                                                                             

Increased salivation                                                                                         

       Number of Animals                    .                     .               7              10     

       Days from ƒ to                       .                     .            15 28          15 27   

                                                                                                             

Noisy respiration                                                                                            

       Number of Animals                    .                     .               .               3     

       Days from ƒ to                       .                     .               .            11 27   

Littering                                                                                                    

       Number of Animals                    1                     .               1               2     

       Days from ƒ to                    38 38                   .            41 41          38 40   

                                                                                                             

Not Observed Due to Littering                                                                                

       Number of Animals                    1                     .               1               2     

       Days from ƒ to                    38 38                   .            41 41          38 40   

                                                                                                             

Scheduled kill                                                                                               

       Number of Animals                   12                    12              12              12     

       Days from ƒ to                    50 53                50 55          50 54          51 64   

                                                                                                             

Increased salivation                                                                                         

       Number of Animals                    .                     2               8              11     

       Days from ƒ to                       .                  47 48          26 51          11 60   

                                                                                                             

Noisy respiration                                                                                            

       Number of Animals                    .                     .               2               4     

       Days from ƒ to                       .                     .            39 48          29 63   

                                                                                                             

Generalised fur loss                                                                                         

       Number of Animals                    .                     .               1               .     

       Days from ƒ to                       .                     .            24 52             .     

Table 2 Group Mean Body Weight Values

Body Weights (g)                                                          

                                                                                                             

                                                                                                                                   

                                                       Day Numbers Relative to Start Date                                        

                                      Group                                                                                       

                                       (Sex)             1      8     15     22     29                                       

                                                                                                                                   

                                     1(M)   Mean     319.1  330.6  343.9  349.4  364.0                                      

                                             S.D.      16.3   18.2   21.2   22.8   24.7                                      

                                             N         12     12     12     12     12                                        

                                                                         

                                     2(M)   Mean     326.9  342.3  358.9  369.3  382.3                                      

                                             S.D.      18.0   19.8   21.1   23.0   26.0                                      

                                             N         12     12     12     12     12                                        

                                                                       

                                     3(M)   Mean     327.3  340.3  355.1  364.3  375.3                                      

                                             S.D.      15.9   18.4   21.6   21.1   25.1                                      

                                             N         12     12     12     12     12                                        

                                                                          

                                     4(M)   Mean     328.5  338.3  344.7  349.0  353.6                                      

                                             S.D.      13.8   14.6   15.5   14.1   13.7                                      

                                             N         12     12     12     12     12                                        

                                     

     

                                                         Body Weights (g)                                                          

                                                                                                                  

                                                                                                                                   

                                                       Day Numbers Relative to Start Date                                        

                                              Group                                                                               

                                               (Sex)             1      8     15                                               

                                                                                                                                   

                                             1(F)   Mean     207.1  211.7  216.7                                              

                                                     S.D.       8.0    9.3    9.1                                              

                                                     N         12     12     12                                                

                                                                                         

                                             2(F)   Mean     209.0  215.3  221.6                                              

                                                     S.D.      10.3   12.2   10.5                                              

                                                     N         12     12     12                                                

                                                                                        

                                             3(F)   Mean     205.3  211.6  218.8                                              

                                                     S.D.       9.4    8.0    9.1                                              

                                                     N         12     12     12                                                

                                                                                          

                                             4(F)   Mean     209.3  212.6  220.5                                              

                                                     S.D.      11.8   12.5   16.6                                              

                                                     N         12     12     12                                                

                                                                 

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/day            

Table 3 Group Mean Body Weight Values - Gestation and Lactation

Group
(Sex)

 

 

 

 

Body Weight (g)

 

 

 

 

Day Numbers

 

 

 

 

 

 

 

 

 

 

 

Gestation

 

Lactation

 

 

 

0

7

14

20

 

1

4

7

13

 

 

 

 

 

 

 

 

 

 

 

 

 

1(F)

 

Mean

 

219.6

241.4

264.2

315.2

 

235.6

249.1

260.9

282.4

 

 

S.D.

 

9.6

12.1

13.9

21.5

 

16.4

16.5

16.6

18.0

 

 

N

 

9

9

9

9

 

9

9

9

9

 

 

 

 

 

 

 

 

 

 

 

 

 

2(F)

 

Mean

 

223.3

247.5

271.8

328.5

 

244.1

264.8

276.9

294.9

 

 

S.D.

 

12.7

13.0

15.6

29.1

 

13.4

17.5

20.6

22.2

 

 

N

 

12

12

12

12

 

11

11

11

11

 

 

 

 

 

 

 

 

 

 

 

 

 

3(F)

 

Mean

 

218.1

240.7

265.6

317.5

 

241.2

256.7

266.7

286.0

 

 

S.D.

 

8.6

10.5

13.9

14.5

 

20.5

13.3

10.6

17.0

 

 

N

 

10

10

10

10

 

10

10

10

10

 

 

 

 

 

 

 

 

 

 

 

 

 

4(F)

 

Mean

 

220.7

239.3

259.6

299.1

 

231.1

234.5

242.3*

261.4*

 

 

S.D.

 

16.4

14.8

16.0

23.3

 

16.0

13.1

14.2

14.2

 

 

N

 

11

11

11

11

 

11

11

11

11

 

 

 

 

 

 

 

 

 

 

 

 

 

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/day                          

  Table 4 Group Mean Body Weight Gains         

                                                                  Increase in Body Weight (g)                                                                 

                                                                                                                                                              

                                                                    Day Numbers Relative to Start Date                                                      

                                                                                              Abs      %                                             

                                                                                              Gain   Gain                                            

                                          Group  From:     1      8       15     22        1      1                                            

                                         (Sex)    To:      8     15       22     29      29     29                                             

                                                                                                                                                       

                                         1(M)    Mean     11.5   13.3      5.5   14.6     44.9   14.0                                          

                                                  S.D.      6.6    5.9      5.1    3.6     14.3    4.3                                          

                                                  N        12     12       12     12       12     12                                            

                                                                                                                                                       

                                         2(M)    Mean     15.3   16.7     10.3   13.0     55.3   16.9                                          

                                                  S.D.      4.3    3.2      7.0    5.3     13.2    3.7                                          

                                                  N        12     12       12     12       12     12                                            

                                                                                                                                                       

                                         3(M)    Mean     13.0   14.8      9.3   10.9     48.0   14.6                                          

                                                  S.D.      4.3    4.7      5.8    7.0     13.5    3.9                                          

                                                  N        12     12       12     12       12     12                                            

                                                                                                                                                       

                                         4(M)    Mean      9.8    6.3**    4.3    4.6**   25.1**  7.7                                          

                                                  S.D.      5.0    6.7      8.2    6.6      8.9    2.8                                          

                                                  N        12     12       12     12       12     12                                            

                                                                                                                                                       

Increase in Body Weight (g)                                                                 

                                                                                                                                                              

                                                                    Day Numbers Relative to Start Date                                                      

                                                                                   Abs      %                                                        

                                                                                   Gain   Gain                                                       

                                                     Group  From:     1     8     1      1                                                       

                                                    (Sex)    To:      8    15   15     15                                                        

                                                                                                                                                       

                                                    1(F)    Mean     4.6   5.0   9.6    4.6                                                      

                                                             S.D.     3.3   4.8   5.2    2.5                                                      

                                                             N       12    12    12     12                                                        

                                                                                                                                                       

                                                    2(F)    Mean     6.3   6.3  12.6    6.1                                                      

                                                             S.D.     6.1   3.3   5.8    2.8                                                      

                                                             N       12    12    12     12                                                        

                                                                                                                                                       

                                                    3(F)    Mean     6.3   7.2  13.4    6.6                                                      

                                                             S.D.     5.4   5.7   7.3    3.7                                                      

                                                             N       12    12    12     12                                                        

                                                                                                                                                       

                                                    4(F)    Mean     3.3   7.9  11.2    5.3                                                      

                                                             S.D.     3.4   6.2   7.2    3.3                                                      

                                                             N       12    12    12     12                                                        

                                                                                                                                                       

Dose Levels: Group 1 ƒ 0(Control)        Group 2 ƒ 100 mg/kg bw/day  Group 3 ƒ 300 mg/kg bw/day  Group 4 ƒ 750 mg/kg bw/day

Table 5 Group Mean Body Weight Gains - Gestation

 

 

Increase in Body Weight(g)

Days

 

 

 

Cumulative Body Weight Change (g)

Days

 

 

 

 

Gestation

 

 

 

Gestation

Group
(sex)

From:

0

7

14

 

 

0

0

To:

7

14

20

 

 

14

20

 

 

 

 

 

 

 

 

 

1(F)

Mean

21.9

22.8

51.0

 

 

44.7

95.7

 

S.D.

4.0

4.1

10.1

 

 

7.2

14.7

 

N

9

9

9

 

 

9

9

 

 

 

 

 

 

 

 

 

2(F)

Mean

24.2

24.3

56.8

 

 

48.4

105.2

 

S.D.

5.1

5.2

18.4

 

 

8.9

23.7

 

N

12

12

12

 

 

12

12

 

 

 

 

 

 

 

 

 

3(F)

Mean

22.6

24.9

51.9

 

 

47.5

99.4

 

S.D.

4.3

4.5

9.4

 

 

7.7

11.5

 

N

10

10

10

 

 

10

10

 

 

 

 

 

 

 

 

 

4(F)

Mean

18.5

20.4

39.5

 

 

38.9

78.4

 

S.D.

4.0

4.6

11.5

 

 

5.5

14.7

 

N

11

11

11

 

 

11

11

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/ 

Table 6 Group Mean Body Weight Gains - Lactation

 

 

Increase in Body Weight (g)
Days

 

 

 

Cumulative Body Weight Change (g)
Days

 

 

 

 

Lactation

 

 

 

Lactation

Group
(sex)

From:

1

4

7

 

 

1

1

To:

4

7

14

 

 

7

14

 

 

 

 

 

 

 

 

 

1(F)

Mean

13.6

11.8

21.6

 

 

25.3

46.9

 

S.D.

9.0

5.7

7.7

 

 

10.2

9.2

 

N

9

9

9

 

 

9

9

 

 

 

 

 

 

 

 

 

2(F)

Mean

20.7

12.1

18.0

 

 

32.8

50.8

 

S.D.

10.5

9.3

5.8

 

 

11.3

12.7

 

N

11

11

11

 

 

11

11

 

 

 

 

 

 

 

 

 

3(F)

Mean

15.5

10.0

19.3

 

 

25.5

44.8

 

S.D.

10.6

6.5

11.6

 

 

13.0

18.8

 

N

10

10

10

 

 

10

10

 

 

 

 

 

 

 

 

 

4(F)

Mean

3.4

7.8

19.1

 

 

11.2*

30.3**

 

S.D.

7.7

6.2

13.4

 

 

10.4

11.3

 

N

11

11

11

 

 

11

11

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/day            

 

Table 7 Group Mean Food Consumptions       

                                                                                                                                                              

                                                             Day Numbers Relative to Start Date                                                              

                                                                                                                                                              

                                                                                                                                                         

                                                             Group From:    1     8                                                                  

                                                            (Sex)   To:     8    15                                                                  

                                                                                                                                                         

                                                            1(M)   Mean    19.9  19.9                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            2(M)   Mean    21.1  21.4                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            3(M)   Mean    20.6  20.5                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            4(M)   Mean    20.0  19.1                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                                                                                                                              

                                                             Day Numbers Relative to Start Date                                                              

                                                                                                                                                              

                                                                                                                                                         

                                                             Group From:    1     8                                                                  

                                                            (Sex)   To:     8    15                                                                  

                                                                                                                                                         

                                                            1(F)   Mean    13.2  14.1                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            2(F)   Mean    15.3  14.8                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            3(F)   Mean    14.3  14.7                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

                                                            4(F)   Mean    13.3  13.9                                                                

                                                                    N       12    12                                                                  

                                                                                                                                                         

Food Consumption = g/animal/day.                                                            

                                                                                                                                                         

Dose Levels: Group 1 ƒ 0(Control)        Group 2 ƒ 100 mg/kg bw/day  Group 3 ƒ 300 mg/kg bw/day  Group 4 ƒ 750 mg/kg bw/day        

                       

Table 8 Group Mean Food Consumptions - Gestation and Lactation

 

 

 

Day Numbers

 

Gestation

 

Lactation

Group
(Sex)

From:

0

7

14

 

1

4

7

To:

7

14

20

 

4

7

13

 

 

 

 

 

 

 

 

 

1(F)

Mean

18.4

19.1

21.3

 

25.0

36.6

48.7

S.D.

2.2

2.3

2.4

 

4.0

4.1

6.2

N

9

9

9

 

9

9

9

 

 

 

 

 

 

 

 

 

2(F)

Mean

18.9

20.2

22.5

 

30.8

40.6

53.8

S.D.

2.0

1.9

2.6

 

7.7

8.0

13.2

N

12

12

12

 

11

11

11

 

 

 

 

 

 

 

 

 

3(F)

Mean

18.3

19.8

21.6

 

34.2

42.0

51.7

S.D.

1.5

1.6

1.4

 

10.9

15.7

9.8

N

10

10

10

 

10

10

10

 

 

 

 

 

 

 

 

 

4(F)

Mean

15.8**

17.3

19.1

 

18.2*

24.4***

35.8**

S.D.

1.8

1.7

1.8

 

3.7

4.5

6.1

N

11

11

11

 

11

11

11

Food Consumption = g/animal/day.                                                           

                                                                                                                                                             

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/day            

Table 9          Food Efficiency

                                                Day Numbers Relative to Start Date                                                

                                                                                                                                   

                                                                                                                             

                                             Group   From:     1       8                                                 

                                             (Sex)     To:     8      15                                                 

                                                                                                                             

                                            1(M)     Mean     8.3     9.6                                                

                                                      N        12      12                                                 

                                                                                                           

                                                                                                                             

                                            2(M)     Mean    10.4    11.1                                                

                                                      N        12      12                                                 

                                                                                                            

                                                                                                                             

                                            3(M)     Mean     9.0    10.3                                                

                                                      N        12      12                                                 

                                                                                                             

                                                                                                                             

                                            4(M)     Mean     7.0     4.8                                                

                                                      N        12      12                                                 

                                                                                                          

                                                Day Numbers Relative to Start Date                                                

                                                                                                                                   

                                                                                                                             

                                             Group   From:     1       8                                                 

                                             (Sex)     To:     8      15                                                 

                                                                                                                             

                                            1(F)     Mean     5.0     4.9                                                

                                                      N        12      12                                                 

                                                                                                           

                                                                                                                             

                                            2(F)     Mean     5.9     6.1                                                

                                                      N        12      12                                                 

                                                                                                            

                                                                                                                             

                                            3(F)     Mean     6.3     7.0                                                

                                                      N        12      12                                                 

                                                                                                             

                                                                                                                             

                                            4(F)     Mean     3.5     8.1                                                

                                                      N        12      12                                                 

                                                                     

Dose Levels: Group 1 ƒ 0 (Control)   Group 2 ƒ 100 mg/kg bw/day   Group 3 ƒ 300 mg/kg bw/day   Group 4 ƒ 750 mg/kg bw/day       

Table 10          Mean Serum T4 concentrations (pg/mL)

Group

Treatment

Dose

(mg/kg bw/day)

Adult terminal males

Male offspring on Day 13 of age

Female offspring on Day 13 of age

1

Control

0#

Mean

42700

39300

42600

SD

11500

7300

7970

CV %

26.9

18.6

18.7

N

12

9

8

2

ELA162

100

Mean

40500

39400

39700

SD

7120

7320

5720

CV %

17.6

18.6

14.4

N

12

11

11

3

ELA162

300

Mean

40400

40100

40000

SD

2950

6910

7980

CV %

7.3

17.2

20.0

N

12

10

10

4

ELA162

750

Mean

34000**

39600

39800

SD

6780

7680

9220

CV %

19.9

19.4

23.2

N

12

11

11

#- Control animals treated with vehicle alone(dried propylene glycol)

Table 11          Mean Serum T3 concentrations (pg/mL) 

Group

Treatment

Dose

(mg/kg bw/day)

Adult terminal males

Male offspring on Day 13 of age

Female offspring on Day 13 of age

1

Control

0#

Mean

660

490

519

SD

211

97

79

N

12

9

8

2

ELA162

100

Mean

795

537

570

SD

153

67

199

N

12

10

10

3

ELA162

300

Mean

784

525

507

SD

130

68

97

N

12

10

9

4

ELA162

750

Mean

659

484

523

SD

214

100

82

N

12

10

9

                                                                                                                                                                   

Conclusions:
Based on this study, a dosage of 300 mg/kg bw/day was considered to represent a No Observed Effect Level (NOEL) and a dosage of 750 mg/kg bw/day was considered to represent the No Observed Adverse Effect Level (NOAEL) for systemic toxicity for both male and female animals.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
750 mg/kg bw/day
Study duration:
subacute
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

Justification for classification or non-classification

Based on the available data, the substance will not be classified.