Fatty acids, C12-14, .alpha.-sulfo, disodium salts

Endpoint summary

Description of key information

OECD TG 408, oral (diet), rat, GLP-conform: NOAEL (females) = 1201 mg/kg bw; NOAEL (males) = 280 mg/kg bw/d

OECD TG 407, oral (diet), rat, GLP-conform: NOAEL (females) = 346 mg/kg bw; NOAEL (males) = 1057 mg/kg bw/d

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

Study period:

05 Jan 2021 - 23 Aug 2021

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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

Specific details on test material used for the study:

Batch No.: 0021197214

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

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

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

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.

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.

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

Key result

Critical effects observed:

no

Endpoint conclusion

Endpoint conclusion:

adverse effect observed

Dose descriptor:

NOAEL

280 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

GLP studies according to OECD guidelines, reliability 1

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 90-day oral toxicity study in Wistar rats

The test item was administered orally via the diet to groups of 10 male and 10 female Wistar rats at concentrations of 0 (test group 0), 1000 (test group 1), 4000 (test group 2) and 12000 ppm (test group 3) over a period of 3 months. Food consumption and body weight were determined weekly. The animals were examined for signs of toxicity or mortality at least once a day. Detailed clinical examinations in an open field were conducted prior to the start of the administration period and weekly thereafter. Clinico-chemical and hematological examinations were performed towards the end of the administration period. After the administration period all animals were sacrificed and assessed by gross pathology. Organ weights were determined followed by histopathological examinations. Stability of the test-substance preparations for a period of 32 days at room temperature, homogeneous distribution of the test substance in the vehicle, and correctness of the prepared concentrations were confirmed by various analyses.

The following test substance-related, relevant findings were noted:

Test group 3: 12000 ppm (874 mg/kg bw/d in males and 1201 mg/kg bw/d in females):

Statistically reduced body weight values in male animals on study days 84 and 91 (-7.7% and -6.9%). Statistically reduced body weight change values in male animals from study days 56-91 (max. -11.8% on study day 70).

No treatment-related, relevant effects were observed among clinical pathology and pathology parameters.

Test group 2 and 1: 4000 ppm (280 mg/kg bw/d in males and 364 mg/kg bw/d in females) and 1000 ppm (69 mg/kg bw/d in males and 81 mg/kg bw/d in females):

Treatment-related, relevant effects were neither observed among clinical examinations, nor among clinical pathology and pathology parameters.

Administration of the test item via the diet to male and female Wistar rats for 3 months caused signs of systemic toxicity in male animals in the high dose group manifested in slight body weight effects. No test substance-related, adverse signs of toxicity were observed in females. Therefore, under the conditions of the present study the no observed adverse effect level (NOAEL) was 4000 ppm (280 mg/kg bw/d) in male and 12000 ppm (1201 mg/kg bw/d) in female Wistar rats.

 

Repeated-dose 28-day oral toxicity study in Wistar rats

In a subacute repeated dose toxicity study according to OECD guideline 407 the test substance was administered via the diet to groups of 5 male and 5 female Wistar rats at concentrations of 0 ppm (test group 0), 1000 ppm (test group 1), 4000 ppm (test group 2) and 12000 ppm (test group 3) in the diet over a period of 4 weeks. Food consumption and body weight were determined weekly. The animals were examined for signs of toxicity or mortality at least once a day. Detailed clinical examinations in an open field were conducted prior to the start of the administration period and weekly thereafter. Beside this, a functional observational battery (FOB) as well as measurement of motor activity (MA) was carried out at the end of the administration period. Clinicochemical and hematological examinations were performed towards the end of the administration period. After the administration period all rats were sacrificed and assessed by gross pathology, followed by histopathological examinations. The administration of the test substance via the diet to male and female Wistar rats for 4 weeks caused treatment-related signs of systemic toxicity at the concentrations of 12000 ppm in the diet of female Wistar rats (1083 mg/kg bw/d) manifested in an anemia, but not in males. Therefore, under the conditions of the present study the no observed adverse effect level (NOAEL) for this test substance was 4000 ppm (346 mg/kg bw/d) in female and 12000 ppm (1057 mg/kg bw/d) in male Wistar rats.

Conclusion

Repeated oral administration of the test item over a period of 3 month caused treatment-related signs of systemic toxicity in males manifested in a slight body weight reduction. Signs of anemia as observed in females at the end of the 28d study could not be confirmed after a administration period of 3 month.

The NOAEL of 280 mg/kg bw/day is therefore considered to be the relevant point of departure for DNEL derivation.

Justification for classification or non-classification

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is not considered to be classified for repeated dose toxicity under Regulation (EC) No 1272/2008, as amended for the seventeenth time in Regulation (EU) 2021/849.