Registration Dossier

Diss Factsheets

Toxicological information

Repeated dose toxicity: oral

Currently viewing:

Administrative data

Endpoint:
sub-chronic toxicity: oral
Type of information:
experimental study
Adequacy of study:
key study
Study period:
05 Jan 2021 - 23 Aug 2021
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2021
Report date:
2021

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)
Version / remarks:
25 Jun 2018
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)
Version / remarks:
30 May 2008
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
BASF SE, Experimental Toxicology and Ecology, Ludwigshafen, Germany
Limit test:
no

Test material

Constituent 1
Chemical structure
Reference substance name:
tetrasodium 2-sulfonatododecanoate 2-sulfonatotetradecanoate
EC Number:
942-523-5
Cas Number:
2156594-77-1
Molecular formula:
Unspecified
IUPAC Name:
tetrasodium 2-sulfonatododecanoate 2-sulfonatotetradecanoate
Test material form:
solid
Specific details on test material used for the study:
Batch No.: 0021197214

Test animals

Species:
rat
Strain:
other: Crl:WI(Han)
Details on species / strain selection:
The rat is a frequently used laboratory animal, and there is comprehensive experience with this animal species. Moreover, the rat has been proposed as a suitable animal species by the OECD and the EPA.
Sex:
male/female
Details on test animals or test system and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories, Research Models and Services GmbH, Sulzfeld, Germany
- Age at study initiation: 41 - 43 days
- Weight at study initiation: The weight variation of the animals used did not exceed 20 percent of the mean weight of each sex.
- Housing: The animals were housed together (5 animals per cage/ sex) in H-Temp polysulfonate cages type 2000P (floor area about 2065 cm2). Motor activity measurements and FOB were conducted in polycarbonate cages type III (floor area about 800 cm2). The cages and wire covers were supplied by TECNIPLAST, Hohenpeissenberg, Germany resp. by Ehret, Emmendingen, Germany. Dust-free wooden bedding was used in this study (the present supplier is documented in the raw data). Wooden gnawing blocks (LIGNOCEL® block large, new name: SAFE® block large), J. Rettenmaier & Söhne GmbH Co KG, Rosenberg, Germany and large play tunnels supplied by PLEXX B.V., Elst, The Netherlands were added for environmental enrichment.
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 8 days

DETAILS OF FOOD AND WATER QUALITY: The food used was ground Kliba maintenance diet mouse/rat “GLP”, meal, supplied by Granovit AG, Kaiseraugst, Switzerland. The supplier assayed the food used in the study for chemical and microbiological contaminants.
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 the presence of microorganisms by a contract laboratory.

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: 13 Jan 2021 To: M: 15 Apr 2021, F: 16 Apr 2021

Administration / exposure

Route of administration:
oral: feed
Details on route of administration:
The oral route was selected since this was proven to be suitable for the detection of a toxicological hazard.
Details on oral exposure:
DIET PREPARATION
- Rate of preparation of diet (frequency): Once at the start of the study
- Mixing appropriate amounts with: For each concentration, the test substance was weighed out and mixed with a small amount of food. In order to obtain the desired concentrations, these premixes were added to the corresponding amounts of food, depending on test group. Mixing was carried out for about 10 minutes in a laboratory mixer.
- Storage temperature of food: Room temperature
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Homogeneity was verified in 3 samples in the highest and lowest concentration (was used as a concentration control at the same time) at the beginning and towards the end of the administration period; additional concentration control analyses were done in the mid concentration. The samples at the beginning were taken from the specific food containers by the staff of the Central Food Mixing unit. The samples towards the end of the administration period were taken from replacement food containers by staff of the testing laboratory at the end of the specific feeding period.
Retain samples for homogeneity/concentration control analyses were taken in the same manner as sampling at the beginning and at the end of the administration period and additionally after 7 weeks.
Duration of treatment / exposure:
90 day
Frequency of treatment:
7 days per week
Doses / concentrationsopen allclose all
Dose / conc.:
1 000 ppm
Remarks:
69 mg/kg bw/d in males and 81 mg/kg bw/d in females
Dose / conc.:
4 000 ppm
Remarks:
280 mg/kg bw/d in males and 364 mg/kg bw/d in females
Dose / conc.:
12 000 ppm
Remarks:
874 mg/kg bw/d in males and 1201 mg/kg bw/d in females
No. of animals per sex per dose:
10
Control animals:
yes, plain diet
Details on study design:
- Dose selection rationale: At the request of the sponsor and based on the findings in the 28d repeated dose toxicity study.
- Rationale for animal assignment: random
- Fasting period before blood sampling for clinical biochemistry: yes, 16 hours

Examinations

Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once a week in the morning
- Cage side observations checked in table No. 1 were included.

BODY WEIGHT: Yes
- Time schedule for examinations: on study day 0 (start of administration period) and thereafter at weekly intervals

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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: at the end of the administration period (study day 91)
- Dose groups that were examined: control and test group 3

HAEMATOLOGY: Yes
- Time schedule for collection of blood:
- Anaesthetic used for blood collection: Yes, isoflurane
- Animals fasted: Yes
- How many animals: all animals per test group
- Parameters checked in table No. 2 were examined.

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: in the morning of day 92/93 (M/F)
- Animals fasted: Yes
- How many animals: all animals per test group
- Parameters checked in table No. 3 were examined.

PLASMA/SERUM HORMONES: Yes
- Time of blood sample collection: in the morning of day 92/93 (M/F)
- Animals fasted: Yes
- How many animals: all animals per test group

URINALYSIS: Yes
- Time schedule for collection of urine: at the end of the administration period (study day 91)
- Metabolism cages used for collection of urine: Yes
- Animals fasted: Yes
- Parameters checked in table No. 5 were examined.

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: at the end of the administration period starting at about 10:00 h
- Dose groups that were examined: all dose groups
- Battery of functions tested: sensory activity, grip strength, motor activity

IMMUNOLOGY: No

OTHER: ESTROUS CYCLE DETERMINATION
Vaginal smears for terminal vaginal cytology examinations were prepared in the morning of the day of sacrifice.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes, see table no. 6
HISTOPATHOLOGY: Yes, see table no. 7
Statistics:
Statistics of clinical pathology
Blood parameters: For parameters with bidirectional changes: Non-parametric one-way analysis using KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the hypothesis of equal medians. For parameters with unidirectional changes: Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians.
Urinalysis parameters (apart from pH, urine volume, specific gravity, color and turbidity): Pairwise comparison of each dose group with the control group using the WILCOXON-test (one-sided) for the hypothesis of equal medians. In case of exactly the same numbers of the dose group and the control, no statistical test is performed.
Urine pH, volume and specific gravity: Non-parametric one-way analysis using KRUSKAL-WALLIS test. If the resulting p-value was equal or less than 0.05, a pairwise comparison of each dose group with the control group was performed using WILCOXON-test (two-sided) for the hypothesis of equal medians.
Urine color and turbidity are not evaluated statistically.

Results and discussion

Results of examinations

Clinical signs:
no effects observed
Description (incidence and severity):
No treatment-related, adverse clinical findings were observed for male and female animals of test groups 1, 2 and 3 (1000, 4000 and 12000 ppm).
One female animal of test group 3 showed a deformed sternum in the region of the processus xiphoideus sterni from study day 65 onwards. This finding was considered as a clinical manifestation of most likely an inherent anomaly which became clinically detectable with a higher body size reached during the study. A treatment related cause of this isolated finding can be excluded.
Mortality:
no mortality observed
Description (incidence):
No animal died during the entire study period.
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
No test substance-related changes of mean body weights and mean body weight change values were observed for male and female animals of test groups 1 and 2 (1000 and 4000 ppm) and in femals animals of test group 3 (12000 ppm). Body weights of male animals of test group 3 (12000 ppm) were statistically reduced on study days 84 and 91 (-7.1% and -6.9%).
Body weight change of male animals of test group 3 (12000 ppm) was statistically reduced from study days 56-91 (max.-11.8% on study day70). These findings were considered as treatment-related and adverse.
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
No test substance-related, adverse changes with regard to food consumption were observed over the entire administration period.
Ophthalmological findings:
no effects observed
Description (incidence and severity):
No treatment-related findings were observed.
All apparent findings were assessed as being incidental in nature since they occurred in control as well as in treated animals and did not show a dose-response relationship.
Haematological findings:
no effects observed
Description (incidence and severity):
No treatment-related changes among hematological parameters were observed.
At the end of the administration period in males of test groups 1 and 3 (1000 and 12000 ppm) absolute monocyte and large unstained cell (LUC) counts were significantly decreased. The alterations were not dose dependent. In males of test groups 2 and 3 (4000 and 12000 ppm) relative basophil counts were significantly increased, but the values were within the historical control range (males, relative basophils 0.1-0.4 %).
In females of test group 2 (4000 ppm) hemoglobin and hematocrit values were significantly decreased, but these alterations were not dose dependent. In females of test group 3 (12000 ppm) total white blood cell (WBC) and absolute lymphocyte counts were significantly increased but the values were within historical control ranges whereas those of the study controls were below these ranges (females, WBC 2.71-4.46 Giga/L, absolute lymphocytes 2.04-3.56 Giga/L).
Therefore, all changes within this chapter were regarded as incidental and not treatment related.
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
No treatment-related, adverse changes among clinical chemistry parameters were observed.
At the end of the administration period, in males of test group 3 (12000 ppm) total protein and globulin values as well as calcium levels were significantly decreased. Total protein and globulin values were within historical control ranges whereas calcium levels were below its
range (males, total protein 59.69-66.25 g/L, globulins 23.49-29.60 g/L, calcium 2.44-2.68 mmol/L). Lower total protein and globulin values were regarded as incidental and not treatment related. Calcium level decreases were the only changed parameter among these individuals.
Therefore, this change was regarded as non-adverse if at all treatment related.
In females of test group 2 (4000 ppm) glucose levels were significantly increased, but this
change was not dose dependent. In females of test group 3 (12000 ppm) LDL-cholesterol values were significantly increased, and the mean was above the historical control range (females, LDL-cholesterol 0.10-0.12 mmol/L). However, this was the only changed parameter among these individuals. LDL-cholesterol in males of this test group was not increased. No changes in thyroid hormone levels were observed in this study. Therefore, LDL-cholesterol increase in females of test group 3 was regarded as non-adverse, if at all treatment related.
Endocrine findings:
no effects observed
Description (incidence and severity):
No treatment-related findings on the estrous cycle were observed.
No treatment-related alterations of T3, T4 and TSH levels were observed.
Urinalysis findings:
no effects observed
Description (incidence and severity):
No treatment-related changes among urinalysis parameters were observed.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
No test substance-related effects were observed in the FOB.
Deviations from "zero values" were obtained in several rats. However, as most findings were equally distributed between test-substance treated groups and controls, were without a dose-response relationship or occurred in single rats only, these observations were considered to have been incidental.
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No treatment-related changes among organ weight parameters were observed.
Absolute mean body weight was statistically significant decreased in males of test group 3 (12000 ppm). However, the decreased terminal (final) body weight was within historical control values and therefore regarded to be no adverse findings.
Relative kidney and liver weights were statistically significant increased and slightly above historical control values in females of test group 3 (12000ppm). However, as the terminal body weight in this test group was decreased (not statistically significant), the absolute organ weights were not significantly changed and no treatment-related findings were observed in microscopy, this organ weight increase was regarded to be unrelated to treatment.
Gross pathological findings:
no effects observed
Description (incidence and severity):
All findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.
Histopathological findings: non-neoplastic:
no effects observed
Description (incidence and severity):
All findings occurred either individually or were biologically equally distributed over control and treatment groups. They were considered to be incidental or spontaneous in origin and without any relation to treatment.

Effect levels

open allclose all
Key result
Dose descriptor:
NOAEL
Effect level:
>= 1 201 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:
280 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male
Basis for effect level:
body weight and weight gain

Target system / organ toxicity

Key result
Critical effects observed:
no

Applicant's summary and conclusion