Registration Dossier

Data platform availability banner - registered substances factsheets

Please be aware that this old REACH registration data factsheet is no longer maintained; it remains frozen as of 19th May 2023.

The new ECHA CHEM database has been released by ECHA, and it now contains all REACH registration data. There are more details on the transition of ECHA's published data to ECHA CHEM here.

Diss Factsheets

Toxicological information

Toxicity to reproduction

Currently viewing:

Administrative data

Endpoint:
screening for reproductive / developmental toxicity
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
From September 19, 2016 to February 17, 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2017

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
other: OECD Guideline 408
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EU Method B.26
Deviations:
no
Qualifier:
according to guideline
Guideline:
other: EPA OPPTS 870.3100
Deviations:
no
GLP compliance:
yes
Limit test:
no

Test material

Constituent 1
Reference substance name:
9-Octadecenoic acid (Z)-, sulfonated, potassium salts
EC Number:
271-843-1
EC Name:
9-Octadecenoic acid (Z)-, sulfonated, potassium salts
Cas Number:
68609-93-8
Molecular formula:
A generic formula cannot be provided for this UVCB substance. The alkyl chain length of the sulfonated fatty acids range from C12-C22, however the major alkyl chain is C18.
IUPAC Name:
9-Octadecenoic acid (Z)-, sulfonated, potassium salts
Test material form:
solid
Details on test material:

Purity/Composition: UVCB (100%);
Appearance: orange-brown crystalline powder with lumps.

Test animals

Species:
rat
Strain:
Wistar
Remarks:
SPF-bred Wistar Han rats
Details on species / strain selection:
Rationale: Recognized by international guidelines as the recommended test system (e.g. EPA, FDA, OECD and EC).
Sex:
male/female
Details on test animals or test system and environmental conditions:
Test system Rat: Crl:WI(Han) (outbred, SPF-Quality).
Source: Charles River Deutschland, Sulzfeld, Germany.
Total number of animals: 40 males, 40 females (females were nulliparous and nonpregnant).
Age at start of treatment: Approximately 6 weeks.
Identification: Earmark and tattoo.
Randomization: By computer-generated random algorithm according to body weight, with all animals within ± 20% of the sex mean.
Acclimatization period: At least 5 days before the start of treatment under laboratory conditions.
Health inspection: Upon receipt of the animals.

Conditions: Environmental controls for the animal room were a mean daily temperature range of 21 to 22°C, a mean daily relative humidity range of 44 to 63%, at least 10 air changes/hour, and a 12-hour light/12-hour dark cycle. The light/dark cycle was interrupted for study related activities.
Accommodation: Group housing of 5 animals per sex in Macrolon cages (MIV type, height 18 cm) with sterilized sawdust as bedding material (Lignocel S 8-15, JRS - J.Rettenmaier & Söhne GmbH + CO. KG) and paper as cage-enrichment (Enviro-dri, Wm. Lilico & Son (Wonham Mill Ltd)). During locomotor activity monitoring, animals were housed individually in a Hi-temp polycarbonate cage (Ancare corp).
Diet: Free access to pelleted rodent diet (SM R/M-Z from SSNIFF Spezialdiäten). During motor activity measurements, animals did not have access to food for a maximum of 1 hour and 43 minutes. Water: Free access to tap water. During motor activity measurements, animals did not have access to water for a maximum of 1 hour and 43 minutes.
Diet, water, bedding and cage enrichment evaluation for contaminants and/or nutrients was performed according to facility standard procedures. There were no findings that could interfere with the study.

Administration / exposure

Route of administration:
oral: gavage
Vehicle:
water
Details on exposure:
Method: Oral gavage, using a plastic feeding tube. Formulations were placed on a magnetic stirrer during dosing. A dose control system (DCS) was used as additional check to verify the dosing procedure according to Standard Operating Procedures.
Dose volume: 5 mL/kg body weight. Actual dose volumes were calculated weekly according to the latest body weight.
Frequency: Once daily, 7 days per week, approximately the same time each day with a maximum of 6 hours difference between the earliest and latest dose.
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
Sampling: Samples (0.5 mL) were taken using a pipette, and were weighed on an analytical balance at 4 decimals precision. During sampling, formulations were placed on a magnetic stirrer. Immediately after sampling (accuracy and homogeneity samples) or after 5 hours at room temperature under normal laboratory light conditions (stability samples), samples were stored on dry ice until receipt.

Analysis: Samples of formulations were analyzed for homogeneity (highest and lowest concentration) and accuracy of preparation (all concentrations). Stability in vehicle over 5 hours at room temperature under normal laboratory light conditions was also determined (highest and lowest concentration). The accuracy of preparation was considered acceptable if the mean measured concentrations were 90-110% of the target concentration. Homogeneity was demonstrated if the coefficient of variation was ≤ 10%. Formulations were considered stable if the relative difference before and after storage was maximally 10%.
Duration of treatment / exposure:
90 d
Frequency of treatment:
daily
Doses / concentrationsopen allclose all
Dose / conc.:
0 mg/kg bw/day (nominal)
Dose / conc.:
100 mg/kg bw/day (nominal)
Dose / conc.:
300 mg/kg bw/day (nominal)
Dose / conc.:
1 000 mg/kg bw/day (nominal)
No. of animals per sex per dose:
10 males and 10 females
Control animals:
yes, concurrent vehicle
Details on study design:
Estrous cycle determination: All females had a daily lavage from Day 70 up to and including Day 90 to determine the stage of estrous (according to Standard Operating Procedures).

Examinations

Parental animals: Observations and examinations:
The following parameters were evaluated: clinical signs daily; functional observation tests in Week 13; body weight and food consumption weekly; ophthalmoscopy at pretest and in Week 13; clinical pathology and macroscopy at termination; organ weights and histopathology on a selection of tissues.
Oestrous cyclicity (parental animals):
Estrous cycle determination All females had a daily lavage from Day 70 up to and including Day 90 to determine the stage of estrous (according to Standard Operating Procedures).

Results and discussion

Results: P0 (first parental generation)

General toxicity (P0)

Clinical signs:
effects observed, treatment-related
Description (incidence and severity):
In the surviving animals, diarrhoea was observed on single occasions in seven 1000 mg/kg animals towards the end of the treatment period. Swelling/gas distention of the abdomen was observed in three 1000 mg/kg and two 300 mg/kg animals, each for single short and transient bouts. Rales were also observed for single short and transient bouts in five 1000 mg/kg animals and two 100 mg/kg animals. No findings were noted during the arena observations in this study.
A dose-related increase in salivation seen after dosing among treated groups was not considered toxicologically relevant, taking into account the nature and minor severity of the effect and its time of occurrence (i.e. after dosing). This sign was considered to be a physiological response related to taste of the test substance rather than a sign of systemic toxicity.
Any other clinical signs noted during the treatment period occurred within the range of background findings.
Mortality:
mortality observed, treatment-related
Description (incidence):
There were two death males treated at 1000 mg/kg.
Body weight and weight changes:
no effects observed
Description (incidence and severity):
Slight (not statistically significant) trends towards a lower body weight and body weight gain were seen in 1000 mg/kg treated animals during a large proportion of the study.
Food consumption and compound intake (if feeding study):
no effects observed
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
not examined
Ophthalmological findings:
effects observed, treatment-related
Description (incidence and severity):
A statistically significant increase in the incidence of focal corneal edema was observed in 100 mg/kg females at Week 13, however this finding was not considered treatment-related as it occurred in a non-dose related manner.
Haematological findings:
effects observed, treatment-related
Description (incidence and severity):
The following (statistically significant) changes in haematology parameters distinguished treated from control animals:
- White blood cell count was increased in 1000 mg/kg females, lymphocyte levels were reduced in 1000 mg/kg males and 1000 mg/kg females, and neutrophil levels were increased in 300 mg/kg males and 1000 mg/kg males and females (males not statistically significant).
- Red blood cell distribution width (%RDW) was increased in 1000 mg/kg females, and haemoglobin levels, mean red blood cell volume (MCV), and mean cell haemoglobin concentration (MCH) were reduced in 1000 mg/kg males and females.
Any other statistically significant changes in haematology parameters were considered to be unrelated to treatment as these occurred in the absence of a dose-related trend.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
The following (statistically significant) changes in clinical biochemistry parameters distinguished treated from control animals:
- Alanine aminotransferase (ALAT) levels were significantly increased in both male and female 1000 mg/kg animals, and aspartate transaminase (ASAT) levels were increased in 1000 mg/kg females.
- Total protein and albumin levels were increased in 1000 mg/kg, and 100/300/1000 mg/kg males, respectively. Total bilirubin levels were increased in 100, 300 and 1000 mg/kg males.
- Urea levels were increased in 1000 mg/kg males and females, and creatinine levels were increased in 100 and 1000 mg/kg males and 1000 mg/kg females.
- Glucose and cholesterol levels both reduced in 1000 mg/kg males and females.
- Inorganic phosphate levels were increased in 1000 mg/kg males and females. Sodium levels were increased in 100 and 300 mg/kg males and females, however these findings occurred in the absence of a dose-related trend and therefore considered not toxicologically relevant.
Any other statistically significant changes in clinical biochemistry parameters were considered to be unrelated to treatment as these occurred in the absence of a dose-related trend.
Urinalysis findings:
not examined
Behaviour (functional findings):
effects observed, treatment-related
Description (incidence and severity):
Foregrip strength was reduced in 300 and 1000 mg/kg males and 1000 mg/kg females in a statistically significant manner. And a slight (not statistically significant) reduction in total movement and ambulatory behavior was observed in 1000 mg/kg females.
Further observations, hearing ability, pupillary reflex, static righting reflex and hindgrip strenght were normal in all examined animals. Motor activity was similar between treated and control males. And all groups showed a similar motor activity habituation profile with a decreasing trend in activity over the duration of the test period.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
effects observed, treatment-related
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Test substance-related microscopic findings were noted, starting at 300 mg/kg in the kidneys and mesenteric lymph nodes of males and females, and at 1000 mg/kg in the adrenal glands of males and females, the pancreas of males, and the spleen of females.
Kidney findings included an increased incidence and severity of tubular vacuolation, with pigmented material in the cortex, which was present in males and females starting at a dose of 300 mg/kg. Tubular degeneration and an increased severity and/or incidence in tubular basophilia was observed at 1000 mg/kg.
In the mesenteric lymph node, an increased incidence and severity of macrophage foci was recorded, starting at 300 mg/kg in both sexes.
In the adrenal gland, vacuolation of the zona glomerulosa was recorded at an increased incidence and/or severity in both sexes at 1000 mg/kg.
In the pancreas, increased apoptosis of acinar cells was recorded in males treated at 1000 mg/kg. One female at 1000 mg/kg with minimal increased apoptosis was considered to be within background levels.
In the spleen, increased incidence and severity of extramedullary hematopoiesis and increased severity of pigmentation was recorded in females treated at 1000 mg/kg.
The remaining histological changes were considered to be incidental findings or within the range of background pathology encountered in rats of this age and strain. There were no test substance-related alteration in the prevalence, severity, or histological character of those incidental tissue alterations.
Histopathological findings: neoplastic:
not examined
Other effects:
effects observed, treatment-related
Description (incidence and severity):
Increased macrophages in male and female mesenteric lymph nodes starting at 300 mg/kg/day, vacuolation of the zona glomerulosa of male and female adrenal glands at1000 mg/kg/day and increased apoptosis in male pancreas at 1000 mg/kg/day were also considered to be non-adverse since these findings were slight in nature.

Reproductive function / performance (P0)

Reproductive function: oestrous cycle:
no effects observed
Description (incidence and severity):
No treatment related changes in estrous cycle length were noted across the dose groups during the period in which estrous cycle length was determined (Day 77 up to and including Day 98).
Reproductive function: sperm measures:
no effects observed
Reproductive performance:
not examined

Details on results (P0)

NOAEL based on on reproductive organs and male and female reproductive system function (e cycle, spermatogenesis)

Effect levels (P0)

Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: estrous cycle regularity, spermatogenesis staging and morphology of gonads and accessory reproductive organs

Results: P1 (second parental generation)

Effect levels (P1)

Key result
Remarks on result:
not measured/tested

Results: F1 generation

Effect levels (F1)

Key result
Remarks on result:
not measured/tested

Overall reproductive toxicity

Key result
Reproductive effects observed:
no
Treatment related:
no

Applicant's summary and conclusion

Conclusions:
Under the study conditions, no toxicologically significant changes were noted in any of the reproductive parameters assessed in this study (estrous cycle regularity, spermatogenesis staging and morphology of gonads and accessory reproductive organs) revealed no treatment-related changes.
Executive summary:

A study was conducted to determine repeated dose toxicity of the test substance when administered to rats according to OECD Guideline 408, EU Method B.26 and EPA OPPTS 870.3100. The test substance, formulated in water, was administered daily for at least 90 d by oral gavage to SPF-bred Wistar Han rats. One control group and three treated groups were tested, each consisting of 10 males and 10 females, at the dose levels of 0, 100, 300 and 1000 mg/kg bw/d. Formulation analyses confirmed that formulations of test substance in water were prepared accurately and homogenously. Under the study conditions, no toxicologically significant changes were noted in any of the reproductive parameters assessed in this study (estrous cycle regularity, spermatogenesis staging and morphology of gonads and accessory reproductive organs) revealed no treatment-related changes (Beerens-Heijnen, 2017).