Registration Dossier

Administrative data

Description of key information

Based on the available weight of evidence, except for reduction in food consumption and body weight changes, no other toxicologically significant or critical systemic effects are expected for the test substance,'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'. However, as a conservative approach,the lowest NOAEL value of 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day; based on read across to isoalkylester study) has been considered further for hazard/risk assessment.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Referenceopen allclose all

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
Combined repeated dose toxicity and reproductive and developmental toxicity study
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
2005
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Remarks:
KL2 due to RA
Justification for type of information:
Refer to section 13 of IUCLID for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Deviations:
not specified
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
- Seventy each of 8-week-old male and female Sprague-Dawley SPF rats (Crj;CD (SD) IGS, Charles River Laboratories Japan, Inc., Atsugi Breeding Center) were purchased (number of animals received: 73 each of males and females)
- The animals were quarantined and acclimatized for 14 days. During this period, the general condition, body weight, and estrous cycle (for 9 days after the quarantine period) were examined, and 58 each of males and females without any abnormality of the general condition (males) (or without any abnormality of the general condition or estrous cycle and with good weight increase for females) were selected and administered the test substance at the age of 10 weeks.
- The body weight at the start of the test substance administration ranged from 338 to 395 g for the males and 219 to 256 g for the females.
- Animals were housed individually in a bracket type metal wire cage in an animal breeding room maintained at a temperature of 21°C to 26°C, relative humidity of 37% to 77%, ventilation frequency of 10 to 15 air changes per hour, and illuminated for 12 hours per day (07:00 to 19:00).
- The animals were allowed free access to Solid chow NMF (nonsterilized: Oriental Yeast Co., Ltd., batch numbers 040713, 040806, 040913) from a stainless steel feeder and to tap water (city water of Gotenba, water bottle was used).
Route of administration:
oral: gavage
Vehicle:
olive oil
Details on oral exposure:
The test substance was administered by oral gavage, a method commonly employed for oral administration to rodents. Each animal received 5 mL/kg body weight of the test suspension by forced oral administration via a gastric tube (08:20-14:24 h). Animals of the control group received the vehicle (olive oil) in a similar manner. The volume of the test suspension was calculated based on the latest body weight of each animal.
Analytical verification of doses or concentrations:
yes
Remarks:
HPLC
Details on analytical verification of doses or concentrations:
- An appropriate amount of the test substance for each concentration was weighed and suspended in olive oil in an agate mortar to prescribed concentrations (50, 100, and 200 mg/mL). The test suspensions were prepared at a frequency of at least once every 7 days and stored in a cold dark place (refrigerator, observed temperature: 3°C-5°C) in light-protected containers (brown glass bottles) until use.
- Stability of the test suspensions was confirmed at Bozo Research Center and showed that 50 and 200 mg/mL suspensions of the test substance (vehicle: olive oil) remained stable for 24 hours at room temperature after storage in light-protected containers in a cold dark place (refrigerator) for 7 days.
- Confirmation of the concentrations and uniformity of the test suspensions of each concentration used for administration at week 1 and on the last week of administration were analyzed by HPLC at the Bozo Research Center. The results showed that for all the suspensions tested, the percentage of the test substance relative to the nominal value was in the range of 96.5% to 105.0%, with a C.V. in the range of 1.0% to 5.3%, which were within the acceptable range (concentration, nominal value ± 10%; C.V., ≤ 10%).
- Analytical method:
The test sample (dosing suspension), 1 mL, was diluted with 60 vol% of THF solution to 10 mL and centrifuged (2000 rpm, 1000 × g, 20°C, 5 minutes); then, 1 mL of the lower layer was diluted to 5 mL with the mobile phase of HPLC. The diluted solution was filtered through a Milex HV filter and the filtrate was subjected to measurement by the HPLC system. Single test samples were taken from the upper, middle and lower layers of the dosing suspension.
Duration of treatment / exposure:
The duration of administration was 14 days before mating, 14 days during the mating period, and 14 days after the mating period, that is, 42 days in total, for the males of the main group and the males and females of the recovery group, and 14 days before mating and up to day 4 of lactation throughout the mating and gestation periods, that is, 42 to 45 days in total, for the females of the main group. The recovery period for the males and females of the recovery group was 14 days after the end of administration, during which period the test substance was not given to the animals.
Frequency of treatment:
Daily
Dose / conc.:
0 mg/kg bw/day (actual dose received)
Dose / conc.:
250 mg/kg bw/day (actual dose received)
Dose / conc.:
500 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
A total of 4 dose groups were set up: 250, 500, and 1000 mg/kg bw groups and the control group.
Each main group consisted of 12 male and female animals each, and each recovery group consisted of 5 each of males and female animals in the control and high- dose groups.
Control animals:
yes, concurrent vehicle
Details on study design:
- Rationale for selection of doses. In a previous study “14-day repeated-dose oral toxicity study of the test substance in rats (a preliminary study)” (doses: 125, 250, 500, and 1000 mg/kg bw, Bozo Research Center study No.: C-R016), administration of the test substance did not produce any effect even at 1000 mg/kg bw, the level defined as the limiting dose by the OECD Guideline for Testing of Chemicals 422. Therefore, 1000 mg/kg was set as the highest dose, and doses of 500 and 250 mg/kg bw were derived by dividing by a common factor of 2. A total of 3 doses were thus set up.

Details on mating procedure
- After the end of the pre-mating administration period, each pair of male and female animals in the same dose group of the main group was housed in the same cage overnight, and the females were judged to have copulated if the vaginal smear contained sperm or the presence of the vaginal plug was confirmed. Days to copulation was calculated from the day of mating, taken as day 0. From gestation day 17 to day 5 of lactation, the animals were housed individually in a plastic Econ cage with
bedding.
- Observation of mother animals. All female animals confirmed to have copulated were allowed to undergo spontaneous delivery, and observed for the presence/absence of abnormalities in the delivery. Delivery completion was checked twice daily (morning, afternoon) from gestation day 21 to the morning of gestation day 25, from which the gestation period was calculated in units of 0.5 days. If delivery was complete by 5:00 h in the afternoon, that day was regarded as day 0 of lactation.
- Mother animals that completed the delivery were observed for the presence/absence of pup licking and ingestion of the placenta and amnion. They were allowed to suckle pups up to day 4 of lactation (the date of delivery completion was regarded as day 0 of lactation) and observed for lactating behavior, using pup gathering, nest building, and breastfeeding as indices.
Positive control:
No
Observations and examinations performed and frequency:
- Observation of the general condition. All animals were observed for the presence of any abnormality of the general condition, such as in the external appearance, nutritional condition, posture, behavior, and excrements, 3 times everyday (before, immediately after, and 2 hours after the administration) during the administration period and once every morning during the recovery period.
Detailed observation of the general condition, function tests, measurement of the grip strength and spontaneous motor activity. Detailed observation of the general condition was performed once before the start of administration for all animals, once every week during the administration period for the males of the main group, once every week during the pre-mating administration and mating periods, and on predetermined days (gestation days 1, 7, 14 and 20, day 4 of lactation) during the gestation period and the lactation period for the females of the main group, and once every week during the administration and recovery periods for the animals of the recovery group. Function tests, measurement of the grip strength, and measurement of the spontaneous motor activity were performed on the last week of administration (day 39 of administration) for the males of the main group, after F1 necropsy on day 4 of lactation (day 42-45 of administration) for the females of the main group, and on the last week of administration (day 39 of administration) and last week of the recovery period (day 11 of recovery) for the males and females of the recovery group. These tests were performed on 5 animals each per group. - Measurement of body weight. Body weight was measured on days 1, 4, 8, 11, 15, 18, 22, 25, 29, 32, 36, 39 and 42 of administration, and on the day of necropsy for the males of the main group, on days 1, 4, 8, 11 and 14 of the recovery period and on the day of necropsy, in addition to the days of measurement for the males of the main group, for the males and females of the recovery group, and on days 1, 4, 8, 11 and 15 of administration (and days 18, 22 and 25 of the administration period as well as in non-copulated animals), days 0, 4, 7, 11, 14, 17 and 20 of gesta tion, and days 0 and 4 of lactation for the females of the main group. Body weight was measured from 08:06 to 10:45 h, except for the measurement on day 0 of lactation for females whose end of delivery was confirmed during the observation in the afternoon. On the day of necropsy, the body weight was measured after the animals had been denied access to food for approximately 16 hours from the previous day, in order to calculate the relative organ weight.
- Measurement of food consumption. The amount of food remaining relative to that supplied on the previous day was measured on days 1, 4, 8, 11, 15, 32, 36, 39 and 42 of administration for the males of the main group, on days 1, 4, 8, 11 and 14, in addition to the days of measurements for the males of the main group, for the males and females of the recovery group, on days 1, 4, 8, 11 and 15 of administration, days 1, 4, 7, 11, 14, 17 and 20 of gestation, and days 2 and 4 of lactation for the females of the main group. Food consumption per animal was calculated from the data thus obtained. The amount of food supplied and the amount of food remaining were measured from 08:26 to 11:25 h.
- Urinalysis (including measurement of water intake). On the last week of administration (day 36 to 37 of administration) and on the last week of the recovery period (day 8 to 9 of recovery), each of the male animals was individually housed in a cage equipped with a urine collector. Four-hour urine specimens were collected under fasting conditions of the animals with free access to water, followed by 20-hour urine specimen collection under free access to food and water. The parameters tested are
as shown below. The first 4-week urine specimens were subjected to tests from pH up to sediments, as well as measurement of the urine volume, and the subsequent 20-hour urine specimens were subjected to measurement of the osmotic pressure and urine volume. Urine volume was calculated as the sum of the volumes of 4-hour and 20-hour urine specimens. The amount of water intake from the previous day was measured using a water bottle while the animals were housed in the cage
equipped with the urine collector.
Sacrifice and pathology:
After delivery, the mother animals were exsanguinated to death by dissecting the abdominal aorta on Day 5 of lactation, after the animals had been denied access to food overnight (approx. 16-20 h) from Day 4 of lactation: 5 in each group after blood collection for hematological tests and blood chemistry tests, and the remaining animals under ether anesthesia.

- Necropsy and organ weight measurement. All of the 5 male and female animals in each group from which blood samples were collected for the hematology and blood chemistry tests on the day after the last day of administration and on the last day of the recovery period were exsanguinated to death after the blood collection, and all the other animals were exsanguinated to death by dissecting the abdominal aorta under ether anesthesia. They were then subjected to detailed gross pathological e
xamination of the body organs and tissues, including the external surface, head, chest and abdomen, and the results were recorded. In the females (mother animals), the number of corpora lutea and number of implantation sites were counted on day 5 of lactation. Then, in 5 each of the male and female animals from which blood samples were collected for the hematology and blood chemistry tests, the weight (absolute) of the following organs (testes and epididymes of all the animals) was measured
and the relative weight of each organ per 100 g body weight was calculated from the absolute organ weight and the body weight at necropsy. For bilateral organs marked with an asterisk, the weight of each side was measured separately and the sum of the weights was calculated. Brain, thyroid gland* (including parathyroid gland), thymus gland, heart, liver, spleen, kidney*, adrenal*, testis*, and epididymis*.

- Histopathological examination. The following organs and tissues of all the animals were fixed and st ored in 10 vol% formalin solution in phosphate buffer (the testes and epididymes were fixed in Bouin's fluid, followed by storage in 10 vol% formalin solution in phosphate buffer). Then, organs and tissues (see below) were embedded in paraffin, and sections were stained with hematoxylin and eosin (H-E). Specimens obtained from 5 each of the male and female animals of the control and high-dose groups
from which blood specimens were collected for the hematology and blood chemistry tests were subjected to microscopic examination (for bilateral organs, both sides were isolated and one side was subjected to the microscopic examination). The results revealed the effect of the test substance on the stomach. Therefore, specimens from 5 each of the male and female animals of the low- and medium-dose groups were also subjected to microscopic examination. Representative cases of normal
and abnormal findings were photographed.

(Cerebrum, cerebellum, pituitary gland, spinal cord (thoracic), sciatic nerve, thyroid gland, parathyroid gland, adrenal, thymus gland, spleen, submandibular lymph nodes, mesenteric lymph nodes, heart, lung (including bronchus), stomach, duodenum, jejunum, ileum, cecum, colon, rectum, liver, kidney, bladder, testis, epididymis, ovary, uterus, seminal vesicle, sternum (including bone marrow), femur (including bone marrow), and the animal identification site (auricle))
Statistics:
- Bartlett test
- Dunnett’s test
- χ2 test with Yates’ continuity correction
- Fisher’s exact test
Clinical signs:
effects observed, non-treatment-related
Description (incidence and severity):
- In the main group, one female in the 500 mg/kg bw group showed opacity of an eyeball (unilateral) from gestation day 5, which was not related to the dose and was therefore considered to be an incidental change.
- In the recovery group, one male in the 1000 mg/kg bw group showed decreased spontaneous motor activity from day 37 of administration up to day 7 of the recovery period, and wheezing from day 37 of administration until the end of the recovery period.
- No abnormality was observed in the other animals, either in the main or in the recovery groups.
Mortality:
no mortality observed
Description (incidence):
- There were no significant difference in the body weight between males and females of the main group. A significantly greater increase in body weight was observed in the females of the 250 and 1000 mg/kg bw groups during the lactation period, but the increase was not dose-related.
- In the recovery group, males in the 1000 mg/kg bw group showed decreased body weight gain during the administration period and decreased body weight (-21g) during the recovery period. This was caused by the abnormality in 1 out of the 5 animals. This animal showed continued body weight decrease (and also decreased spontaneous activity and wheezing in the observation of the general condition; the body weight was 466g before manifestation of the symptom and 261g on day 14 of the recovery period). The body weights of the other 4 males and 5 females in the same group were similar to those of the animals in the control group, showing no statistically significant differences.
Body weight and weight changes:
effects observed, non-treatment-related
Food consumption and compound intake (if feeding study):
effects observed, non-treatment-related
Description (incidence and severity):
Administration of the test substance did not have any effect on the food consumption in the males or females of either the main group or the recovery group. A significant increase was observed on day s 2 and 4 of lactation in the females of the 250 mg/kg dose group in the main group, but this was not dose-related.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
not examined
Haematological findings:
effects observed, non-treatment-related
Description (incidence and severity):
-Tests at the end of the administration period. A significant increase in the serum level of fibrinogen was observed in the females of the 250 mg/kg bw group and a significant decrease in the percentage of lymphocytes and significant increase in the percentage of segmented neutrophils were observed in the females of the 500 mg/kg bw group. However, none of these changes were observed in the 1000 mg/kg bw group, suggesting that they were within the range of physiological variations. No significant differences were observed in the male animals between the control group and any of the treatment groups.
- Tests at the end of recovery period. A significant increase in the mean corpuscular volume of the red blood cells, significant decrease of the platelet count, significant increase in the percentage of lymphocytes, and significant decrease in the percentage of segmented neutrophils were observed in females of the 1000 mg/kg bw group. However, these changes were not observed at the end of the administration period, which suggested that they were within the range of physiological variations. No significant differences were observed in the male animals between the control group and any of the treatment groups.
Clinical biochemistry findings:
effects observed, non-treatment-related
Description (incidence and severity):
- Test at the end of the administration period. A significant increase in the serum level of ALT was observed in the males of the 1000 mg/kg bw group. A significant decrease of the serum level of inorganic phosphorus was observed in the males of the 250 mg/kg bw group. However, since the decrease was not dose-related, it was considered to be within the range of physiological variations.
- Tests at the end of recovery period. A significant increase in the serum level of total protein was observed in the females of the 1000 mg/kg bw group. However, since no such change was observed at the end of the administration period, the increase was considered to be within the range of physiological variations.
Urinalysis findings:
no effects observed
Description (incidence and severity):
Urinalysis (including measurement of water intake) showed no abnormalities in the qualitative parameter values in any of animals in either the main group or in the recovery group. No significant difference was observed in any of the quantitative parameter values between the control group and any of the treatment groups.
Behaviour (functional findings):
no effects observed
Description (incidence and severity):
- Observation of animals in the home cage. No abnormalities were observed in any of the animals in either the main group or the recovery group.
- Observation of the animals while being handled. No abnormality was observed in any of animals in either the main group or the recovery group.
- Observation of animals in the open field. Males of the 1000 mg/kg bw group in the main group showed a significant increase of the defecation frequency during weeks 1 and 2 of administration, which was a very mild transient change and considered to be within normal range. No abnormalities were observed in the other parameters in any of the animals in either the main group or the recovery group. No significant differences were observed in the standing frequency between the control group and any of the treatment groups.
- Function tests. No abnormalities were observed in any of the animals in either the main group or the recovery group. No significant differences were observed in the air righting reflex or landing foot splay between the control group and any of the treatment groups.
- Measurement of the grip strength. No significant differences were observed between the control group and any of the treatment groups in either the main group or the recovery group.
- Measurement of spontaneous motor activity (measured for 10-minute periods and a total of 60 minutes). No significant difference was observed between the control group and any of the treatment groups in either the main group or the recovery group.
Immunological findings:
not examined
Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
No dose-related changes in either direction (increase or decrease) were observed in either the absolute or the relative weight. Although significant differences in the weights of the following organs were observed, they were considered to be within the range of normal variations, because they were neither dose-related nor were observed at the end of the administration period.
Gross pathological findings:
effects observed, treatment-related
Description (incidence and severity):
- Findings at the end of the administration period. Stomach: Indentation of the anterior stomach was observed in 0, 5 and 7 males, and 1, 1 and 3 females of the 250, 500 and 1000 mg/kg bw groups, respectively. White foci were observed in 1 male of the 500 mg/kg bw group. Rough mucosa in the anterior stomach was observed in 5 and 9 males, and 5 and 6 females of the 500 and 1000 mg/kg bw groups, respectively. Indentation of the glandular stomach was observed in 1 female of the 500 mg/kg bw group. All the other findings observed in the organs and tissues were considered to be incidental, as judged from the frequency of their occurrence and the pathological findings (Dark red foci in the glandular stomach were observed in 0, 1, 2 and 1 males and 4, 2, 1 and 1 females of the control
group, 250, 500 and 1000 mg/kg bw groups, respectively. Kidney: Pyelectasis was observed in 2 and 1 males of the 250 and 1000 mg/kg bw groups, respectively. Eyeballs: Corneal opacity (unilateral) was observed in 1 female of the 500 mg/kg bw group).
- Findings at the end of the recovery period. Stomach: Rough mucosa in the anterior stomach was observed in 1 male of the 1000 mg/kg bw group. This animal also showed expansion of the digestive tract from the stomach to the colon due to gas accumulation, and a mild decrease in the size of the testis. Other findings observed in the following organs were considered to be incidental as judged from the frequency of their occurrence and the pathological findings (Lung: Dark red foci were observed in 1 female of the 1000 mg/kg bw group).
Neuropathological findings:
not examined
Histopathological findings: non-neoplastic:
effects observed, treatment-related
Description (incidence and severity):
Administration of the test substance had effects on the stomach of the animals in the 250 mg/kg bw and higher dose groups.
- Findings at the end of the administration period.
Stomach: Mild to moderate erosions or ulcers of the anterior stomach were observed in 0, 4 and 4 males and 1, 1 and 1 females of the 250, 500, and 1000 mg/kg bw groups, respectively. Very mild to moderate thickening of the anterior stomach mucosa was observed in 1, 4 and 5 males and 1, 4 and 3 females of the 250, 500, and 1000 mg/kg bw groups, respectively. Very mild to mild edema of the submucosal tissue in the anterior stomach was observed in 1, 5 and 5 males and 0, 4 and 3 females of the 250, 500, and 1000 mg/kg bw groups, respectively. Most of these changes in the anterior stomach were localized findings. All of the other findings observed, as follows, were considered to be incidental changes as judged from the frequency of their occurrence and the histopathological findings
Epididymis: Very mild infiltration by stromal cells was observed in 1 male of the control group. Heart: Very mild localized myocarditis was observed in 4 males of the control group and 1 male of the 1000 mg/kg bw group.
Kidney: Very mild basophilic tubules were observed in 3 males of the control group and 1 male and 1 female in the 1000 mg/kg bw group.
Liver: Very mild, minute granulomas were observed in 3 males of the control group and 1 male of the 1000 mg/kg bw group.
Lung (including bronchi): Very mild mineral deposits in the arterial walls were observed in 1 male of the control group and 1 female of the 1000 mg/kg bw group. Very mild accumulation of foam cells was observed in 2 males and 1 female of the control group, and 1 male and 3 females of the 1000 mg/kg bw group.
Spleen: Very mild to mild extramedullary hematopoiesis was observed in 5 females each in the control group and 1000 mg/kg bwgroup.
Stomach: Inclusion cysts were observed in 1 male of the 500 mg/kg bw group. Very mild to mild erosions in the glandular stomach were observed in 0, 0, 0 and 1 male and 3, 1, 1, and 0 females in the control group, 250, 500, and 1000 mg/kg bw group, respectively).

- Findings at the end of the recovery period. Stomach: Moderate thickening of the anterior stomach mucosa was observed in 1 male of the 1000 mg/kg bw group.
Histopathological findings: neoplastic:
no effects observed
Other effects:
no effects observed
Description (incidence and severity):
Refer to the other related RSS for details on reproductive and development toxicity.
Dose descriptor:
NOAEL
Effect level:
ca. 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No systemic effects observed up to 1000 mg/kg bw/d; the changes in forestomach are of local nature due to the irritant properties of the substance
Key result
Dose descriptor:
LOAEL
Effect level:
ca. 250 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: effects on forestomach
Key result
Critical effects observed:
no
Conclusions:
Based on the results of the read across study, NOAEL for systemic effects for the test substance, is considered to be 1000 mg/kg bw/day.
Executive summary:

A screening study was conducted to determine the toxicity to reproduction of the read across substance, 'mono- C12 PSE, Na+' (purity: 100%, according to the OECD Guideline 422, in compliance with GLP. The read across substance was administered at 0 (control group), 250, 500 or 1000 mg/kg bw to male Sprague-Dawley SPF rats for 14 days before mating, through the mating period, and up to 1 d before necropsy (42 d in total) and to female Sprague-Dawley SPF rats for 14-d before mating, through the mating period and the gestation period, up to Day 4 of lactation (42 to 45 d in total) to investigate the repeated-dose, reproductive and developmental toxicities. In the 0 and 1000 mg/kg bw groups, a 14-d recovery period was allowed after the 42-d administration period to investigate the reversibility of the toxic changes. Administration of the read across substance had no effect on any of the following: general condition, findings on detailed observation of the general condition, results of function tests, grip strength, spontaneous motor activity, body weight, food consumption, results of urinalysis (including water intake), or in the results of haematological or blood chemistry tests. Administration of the read across substance had no effect on any of the following: general condition, findings on detailed observation of the general condition, results of function tests, grip strength, spontaneous motor activity, body weight, food consumption, results of urinalysis (including water intake), or in the results of haematological or blood chemistry tests. Changes in the forestomach may be considered to be adverse, however, they are also considered to be a result of local irritation of the (irritant) test substance (which is brought directly in contact to the mucosa in a massive amount by gavage application) than a true effect of systemic toxicity. Furthermore, in the light of the absence of a forestomach in humans, observed effects on this tissue are of questionable relevance with reference to the extrapolation of the toxic properties of a read across substance in humans. Under the study conditions, only (local) changes of the forestomach mucosa were observed at 250 mg/kg bw/day and above, which are due to the irritant properties of the substance. Therefore, the NOAEL (systemic) is considered to be 1000 mg/kg bw/day (BRC, 2005). Based on the results of the read across study, a similar systemic NOAEL can be expected for the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

Endpoint:
short-term repeated dose toxicity: oral
Type of information:
experimental study
Adequacy of study:
weight of evidence
Study period:
1985
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 407 (Repeated Dose 28-Day Oral Toxicity in Rodents)
Deviations:
not specified
GLP compliance:
yes
Limit test:
no
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
Test organism
- Age/Weight at study initiation: M 84-98 g; F 81-93g


Route of administration:
oral: gavage
Details on route of administration:
- Oral gavage
- Concentration in vehicle: 0, 2, 10 or 20%
- Total volume applied: 5 mL/kg
Vehicle:
olive oil
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
- Doses: 0, 100, 500 and 1000 mg/kg/day actual received
Duration of treatment / exposure:
27 - 28 d exposure
Frequency of treatment:
5 d/week
Remarks:
Concentration in vehicle: 0, 2, 10 or 20% test substance
Remarks:
Doses: 0, 100, 500 and 1000 mg/kg/day actual dose received
No. of animals per sex per dose:
10M + 10F per dose level plus 5M+5F per dose level for reversibility
Control animals:
yes, concurrent vehicle
Details on study design:
- Post exposure period: 28 d
Observations and examinations performed and frequency:
- Clinical signs: Daily
- Mortality: Daily
- Body weight: Weekly
- Food consumption: Daily
- Water consumption: Weekly
- Ophthalmoscopic examination: At end of study
- Haematology: After 21/22 daily doses: Haematocrit, MCV, Hb, RBC, WBC, Thrombocytes, differential white count
- Biochemistry: After 21/22 daily doses: Serum Urea, creatinine, Na, K, calcium, alkaline phosphatase, ALAT, ASAT, GT, bilirubin, chloride, albumin, total protein, chloesterol
- Urinalysis: Not done
Sacrifice and pathology:
Organs examined at necropsy: Macroscopy and microscopy
- Macroscopic: Yes
- Organs weights: thyroid, adrenals, thymus, kidney, spleen, heart, brain, testes, liver
- Microscopic: All organs from the control and top dose animals were examined plus the animals from the reversibility study
Statistics:
- T-test
- U-test for organ weights
Clinical signs:
no effects observed
Mortality:
no mortality observed
Description (incidence):
Comparable with controls
Body weight and weight changes:
no effects observed
Food consumption and compound intake (if feeding study):
no effects observed
Description (incidence and severity):
Similar to control group
Ophthalmological findings:
no effects observed
Description (incidence and severity):
Comparable in treated and control animals
Haematological findings:
no effects observed
Description (incidence and severity):
No differences between treated and control animals other than an increase in neutrophils containing rodlike bodies observed in top dose females (confidence level 95%*). Values obtained (% rod like cells) were 2.5 (Control), 3.3 (low dose), 2.9 (mid dose), 5.3* (high dose).
Clinical biochemistry findings:
no effects observed
Description (incidence and severity):
Statistically significant changes (*95% ** 99% confidence) in some clinical chemical parameters were noted as follows:
a) 500 mg/kg/day males increased potassium*
b) 500 mg/kg/day females increased GGT*, cholesterol** and chloride*
c) Glucose was elevated in top dose males (1000 mg/kg/day)** : Serum glucose mmol/L: Control (6.03), low (6.20), mid (6.25) and high (7.28**)

These changes were not dose and/or sex related and not correlated with any histopathological findings and are therefore not considered of toxicological significance.

Organ weight findings including organ / body weight ratios:
no effects observed
Description (incidence and severity):
Both absolute and relative organ weights were essentially comparable in treated and control animals.
Sporadic changes were observed as follows (*95% ** 99% confidence)
1) increases in absolute organ weight: (a) male kidney 500 mg/kg/day* (b) male testes 1000 mg/kg/day*.
The only change in relative organ weight was an increase in male adrenal weight at 1000 mg/kg/day*.

- Testes weight mean relative (absolute):
a) Control: 0.856 (3.207)
b) Low: 0.839 (3.186)
c) Mid: 0.908 (3.455)
d) High: 0.893 (3.474*)

- Adrenal weight mean relative (absolute)
a) Control: 0.013 (0.050)
b) Low: 0.014 (0.054)
c) Mid: 0.014 (0.055)
d) High: 0.015* (0.058)
Histopathological findings: non-neoplastic:
effects observed, non-treatment-related
Description (incidence and severity):
Histopathological changes in test, control or reversibility groups
Key result
Dose descriptor:
NOAEL
Effect level:
> 1 000 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
male/female
Basis for effect level:
other: No treatment related observed effects observed
Remarks on result:
other: i.e., equivalent to >950 mg a.i./kg bw/day
Critical effects observed:
no
Conclusions:
Under the study conditions, the NOAEL for systemic effects was determined to be >1000 mg/kg bw/day (i.e., >950 mg a.i./kg bw/day).
Executive summary:

A 28-d study was conducted to determine the sub-acute toxicity of the test substance, hexadeacn-1-ol (purity: >95%) in Sprague-Dawley rats, according to a method similar to OECD Guideline 407, in compliance with GLP. In the study, SD rats (10 each sex/dose + 5 each sex/dose for reversibility) were exposed to the 0, 2, 10 and 20% test substance in olive oil, 5mL/kg bw by oral gavage for 28 d, 5d/week. The actual doses received were calculated to be 0, 100, 500 and 1000 mg/kg bw/day respectively. Clinical signs, mortality and food consumption were observed daily, whereas, body weight and water consumption were analysed weekly. Opthalmoscopic examination was performed at the end of the study. Biochemistry and haematology analysis were performed 21/22 d after daily dosing. There were no treatment related effects on clinical signs, body weight, mortality and food consumption were noted. In haematology analysis, no differences between treated and control animals other than an increase in neutrophils containing rod like bodies observed in top dose females (1000 mg/kg bw/day) were noted. Statistically significant changes in some clinical chemical parameters were noted, where at 500 mg/kg/day, males increased potassium and females increased GGT, cholesterol and chloride, glucose was elevated in top dose males (1000 mg/kg/day). Both absolute and relative organ weights were essentially comparable in treated and control animals. Increases in absolute weight in male kidney at 500 mg/kg/day and male testes at 1000 mg/kg/day were noted. The only change in relative organ weight was an increase in male adrenal weight at 1000 mg/kg/day. Sporadic statistically significant changes in some organ weights and clinical chemical parameters were observed but these were not associated with histopathological changes and were not considered of toxicological significance. Under the study conditions, NOAEL was considered to be >1000 mg/kg/day (i.e., >950 mg a.i./kg bw/day), based on lack of toxicologically significant treatment related effects at this dose level (top dose level) (OECD SIDS, 2006).

Endpoint:
short-term repeated dose toxicity: oral
Remarks:
combined repeated dose and reproduction / developmental screening
Type of information:
read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Study period:
From November 01, 2012 to December 13, 2012
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study
Remarks:
KL2 due to RA
Justification for type of information:
Refer to section 13 of IUCLID for details on the read-across justification. The study with the read across substance is considered sufficient to fulfil the information requirements as further explained in the provided endpoint summary.
Reason / purpose:
read-across source
Qualifier:
according to
Guideline:
OECD Guideline 422 (Combined Repeated Dose Toxicity Study with the Reproduction / Developmental Toxicity Screening Test)
Version / remarks:
22 Mar 1996
Deviations:
no
GLP compliance:
yes (incl. certificate)
Remarks:
Behoerde fuer Soziales, Familie, Gesundheit und Verbraucherschutz; Hamburg, Germany
Species:
rat
Strain:
Crj: CD(SD)
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Laboratories Research Models and Services Germany GmbH, Sulzfeld, Germany
- Age at study initiation: males: 50 days; females: 60 days
- Mean weight at study initiation: males: 248.8 to 298.7 g; females: 195.2 to 228.1 g
- Fasting period before study: no
- Housing: single housing in MAKROLON cages (type III plus)
- Diet: commercial ssniff R-Z V1324 (ssniff Spezialdiäten GmbH, Soest, Germany); ad libitum
- Water: tap water; ad libitum
(Analyses of diet and water was performed.)
- Acclimation period: 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 3
- Humidity (%): 55 ± 15
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
corn oil
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 50, 150 and 500 mg/mL
- Amount of vehicle (if gavage): 2 mL/kg
Analytical verification of doses or concentrations:
yes
Details on analytical verification of doses or concentrations:
For the analysis of the test item-vehicle mixtures samples were taken at the following time points and stored at ≤ -20°C until analysis:
Start of treatment period; immediately after preparation of the test item-vehicle mixtures; 8 and 24 hours after storage of the test item preparations at room temperature; end of treatment period; during treatment with the test item always before administration to the last animal of the dose level group
The following parameters were determined: linearity, accuracy, precision, sensitivity, specificity, stability at +2°C to +8°C or -20°C (0, 24, 72 and 168 hours)
Duration of treatment / exposure:
Males: The daily administration of the test substance was started two weeks before mating and lasted until test day 35, which was one day before sacrifice.
Females: The daily administration of the test substance was started two weeks before mating and continued to at least day 3 of lactation.
Maximum: 56 d of treatment
Frequency of treatment:
once daily; 7 days/week
Dose / conc.:
100 mg/kg bw/day (actual dose received)
Dose / conc.:
300 mg/kg bw/day (actual dose received)
Dose / conc.:
1 000 mg/kg bw/day (actual dose received)
No. of animals per sex per dose:
10
Control animals:
yes, concurrent vehicle
Details on study design:
- Dose selection rationale: based on a 14-day range-finding study (Leuschner, 2012)
Observations and examinations performed and frequency:
CAGE SIDE OBSERVATIONS: Yes
- Time schedule: before and after dosing
- Cage side observations checked: skin/fur, eyes, mucous membranes, respiratory and circulatory systems, somatomotor activity and behaviour patterns

MORTALITY AND CLINICAL SIGNS
- Time schedule: at least once daily (the frequency was increased when signs of toxicity were observed); deaths were recorded twice daily (animals which died or were sacrificed during the study were necropsied as soon as possible after exitus)

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: once before the first exposure (to allow within-subject comparisons) and once a week thereafter
- Paramters: changes in skin, fur, eyes, mucous membranes, occurrence of secretions and excretions and autonomic activity (e.g. lacrimation, pilo-erection, pupil size, and unusual respiratory pattern); changes in gait, posture and response to handling as well as the presence of clonic or tonic movements, stereotypies (e.g. excessive grooming, repetitive circling), difficult or prolonged parturition or bizarre behaviour (e.g. self-mutilation, walking backwards)

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: Yes
- Compound intake calculated as time-weighted averages from the consumption and body weight gain data: No data

FOOD EFFICIENCY:
- Body weight gain in kg/food consumption in kg per unit time X 100 calculated as time-weighted averages from the consumption and body weight gain data: No

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study): Yes
- Time schedule for examinations: daily by visual appraisal

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations:
- Dose groups that were examined: all dose groups

HAEMATOLOGY: Yes
- Time schedule for collection of blood: at the end of the pre-mating period
- Anaesthetic used for blood collection: Yes (ether)
- Animals fasted: Yes, overnight
- How many animals: 5/sex/dose
- Parameters: haemoglobin, erythrocytes, leucocytes, differential blood count (absolute/relative), reticulocytes, platelets, haematocrit, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, thromboplastin time, activated partial thromboplastin time

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at the end of the pre-mating period
- Animals fasted: Yes, overnight
- How many animals: 5/sex/dose
- Parameters examined: albumin, globulin, albumin/globulin ratio, bile acids, bilirubin, cholesterol (total), creatinine, glucose, urea, total protein, calcium, chlorid, potassium, sodium, alanine aminotransferase (ALAT), alkaline phosphatase (AP), aspartate aminotransferase (ASAT)

URINALYSIS: No

NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: two hours after dosing shortly before scheduled sacrifice in 5 males per group (day 35); two hours after dosing during lactation, shortly before scheduled sacrifice in 5 females per group (day 39-56)
Screening of assessment were conducted as described on the following pages in five males and five females randomly selected from each group.
- Dose groups that were examined: all dose groups
- Battery of functions tested: sensory reactivity to stimuli of different types (e.g. auditory, visual and proprioceptive stimuli) (based on Gad), as well as the assessment of grip strength (Meyer) and motor activity

OTHER: Qualitative sperm analysis was performed.
(for further reproduction parameters see respective study entry (chapter 7.8.1))
Males were sacrificed on day 36, females were sacrifices on day 4 post-partum or shorty thereafter.
Sacrifice and pathology:
GROSS PATHOLOGY: Yes
- Organ weights: epididymes and testicles (all males); adrenal gland, brain, heart, kidney, liver, spleen, thymus (5/sex/dose)
- Fixation: epididymis, gross lesions, mammary gland, ovary, prostate, seminal vesicle, testicle, uterus (incl. cervix and oviducts), vagina (all animals); adrenal gland, bone marrow (os femoris), brain (cerebrum, cerebellum, brain stem), heart (left and right ventricle, septum), intestine, small (duodenum, jejunum, ileum, incl. Peyer's patches, Swiss roll method), intestine, large (colon, rectum), kidney and ureter, liver, lungs (with mainstem bronchi and bronchioles), preserved by inflation with fixative and then immersion, lymph node (1 cervical, 1 mesenteric), nerve (sciatic), oesophagus, spinal cord (3 sections), spleen, stomach, thyroid (incl. parathyroids), thymus, tissue masses or tumours (incl. regional lymph nodes), tongue (incl. base), trachea (incl. larynx), urinary bladder (5/sex/dose)
HISTOPATHOLOGY: Yes, all organs that were included for fixation (5/sex of control and high dose group)
Statistics:
STUDENT's t-test (p ≤ 0.01): all numerical functional tests
Multiple t-test based on DUNNETT (p≤0.05 and p ≤ 0.01): body weight, food consumption, haematology, clinical chemistry, absolute and relative organ weights
For all numerical values homogeneity of variances was tested by using the BARTLETT chi-square test. If the variances were homogeneous, the DUNNETT test (p ≤ 0.01) was used to compare the experimental groups with the control group. In case of heterogeneity of variances, the STUDENT's t-test was carried out; limit of significance was p ≤ 0.01.
Clinical signs:
no effects observed
Mortality:
no mortality observed
Body weight and weight changes:
effects observed, treatment-related
Description (incidence and severity):
statistically significant decrease (-9.4%) in body weight in females during lactation at 1000 mg/kg bw/day
Food consumption and compound intake (if feeding study):
effects observed, treatment-related
Description (incidence and severity):
In the high dose group (females: gestation/lactation period) a statistically significant reduction in food consumption by 21.7% was noted.
Food efficiency:
not examined
Water consumption and compound intake (if drinking water study):
no effects observed
Ophthalmological findings:
no effects observed
Haematological findings:
no effects observed
Clinical biochemistry findings:
no effects observed
Urinalysis findings:
not examined
Behaviour (functional findings):
no effects observed
Organ weight findings including organ / body weight ratios:
no effects observed
Gross pathological findings:
no effects observed
Histopathological findings: non-neoplastic:
no effects observed
Histopathological findings: neoplastic:
not examined
Details on results:
CLINICAL SIGNS AND MORTALITY
Piloerection was seen in 1 female of the high dose group at on day 2-4 of lactation.

BODY WEIGHT AND WEIGHT GAIN
A reduction in body weight (-9.7%) in high dose females during lactation period.

FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study)
Reduction in food intake (-21.7%) in high dose females during gestation/lactation.

WATER CONSUMPTION AND COMPOUND INTAKE (if drinking water study)
No effects on water consumption in any treatment group were observed.

OPHTHALMOSCOPIC EXAMINATION
No effects were observend in any treatment group.

HAEMATOLOGY
No effects on water consumption in any treatment group were observed.

CLINICAL CHEMISTRY
An decrease of ASAT activity was seen in high dose females on day 15. This effects was considered to be due to the relative low or high value observed for the control group and not to be test item related.

NEUROBEHAVIOUR
No effects observed.

ORGAN WEIGHTS
All effects observed (slight increase in absolute and relative liver weight in males) were still within the historical control data of the laboratory and thus not of toxicological relevance.

GROSS PATHOLOGY
No effects observed.

HISTOPATHOLOGY:
No effects observed.

OTHER FINDINGS
The qualitative sperm staging revealed no test item-related specific spermatogenic changes in the male animals from the high dose group (1000 mg/kg b.w./day).
Key result
Dose descriptor:
NOAEL
Effect level:
300 mg/kg bw/day (actual dose received)
Based on:
test mat.
Sex:
female
Basis for effect level:
other: adverse effects on body weight and body weight gain and food consumption
Remarks on result:
other: i.e., equivalent to 233.4 mg ai./kg bw/day
Key result
Dose descriptor:
NOAEL
Effect level:
1 000 mg/kg bw/day (nominal)
Based on:
test mat.
Sex:
male
Basis for effect level:
other: no adverse affects observed
Remarks on result:
other: i.e., equivalent to 778 mg ai./kg bw/day
Key result
Critical effects observed:
not specified

Individual body weights of females during the pre-mating and lactation period.

Control group

Day(s) relative to start

 

1

8

15

11

196

209

212

12

217

228

243

13

210

220

213

14

217

234

244

15

3211

215

232

16

195

208

197

17

205

224

239

18

212

238

233

19

224

236

259

20

200

218

214

Mean

209

223

229

SD

9

10

19

1000 mg/kg bw

Day(s) relative to start

 

1

8

15

71

200

210

225

72

210

231

234

73

206

234

247

74

210

222

235

75

194

219

218

76

218

243

223

77

214

230

227

78

222

225

210

79

198

200

201

80

203

223

231

Mean

207

224

225

SD

9

12

13

Control group

Day(s) relative to littering

 

 

1

4

11

286

309

12

323

330

13

325

311

14

331

327

15

311

322

16

282

302

17

309

327

18

312

328

19

315

331

20

300

329

Mean

309

322

SD

16

10

1000 mg/kg bw

Day(s) relative to littering

 

 

1

4

71

270

281

72

339

301

73

289

309

75

289

317

77

253

247

78

280

271

79

290

296

80

293

308

Mean

288

291

SD

24

23

Relative food consumption of females between day 1 and 4 of lactation

Control group

1000 mg/kg bw

 

 

11

122

71

115

12

91

72

96

13

105

73

63

14

107

75

110

15

106

77

30

16

121

78

63

17

114

79

98

18

100

80

110

19

101

 

 

20

126

 

 

Mean

109

Mean

86

SD

11

SD

30

Conclusions:
Under the read across study conditions, the NOAEL for systemic toxicity was determined to be 300 mg/kg bw/day (i.e., equivalent to 233.4 mg ai./kg bw/day) in females and 1000 mg/kg bw/day (i.e., equivalent to 778 mg a.i./kg bw/day) in male rats in this study.
Executive summary:

A study was conducted to determine the repeated dose toxicity of the read across substance, 'Isodecyl oleate' (purity: 77.8%), according to OECD Guideline 422, in compliance with GLP. Three groups of ten male and ten female Sprague-Dawley rats received the read across substance daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until Day 3 post-partum. The read across substance was administered as a suspension in the vehicle, corn oil, at dose-levels of 0, 100, 300 or 1000 mg/kg bw/day. Another group of ten males and ten females received the vehicle alone, under the same experimental conditions and acted as a control group. The concentration of the dose formulation was checked start of treatment period; immediately after preparation of the read across substance-vehicle mixtures; 8 and 24 h after storage of the read across substance preparations at room temperature; end of treatment period; during treatment with the read across substance always before administration to the last animal of the dose level group. The animals were checked at least twice daily during the dosing period for mortality and morbidity and once daily for clinical signs. Detailed clinical observations were performed once a week. Body weight and food consumption were recorded once a week during premating and mating periods (food consumption not during mating), and during gestation on post coital and lactation on Days 1 and 4 post-partum. The animals were paired for mating after 2 weeks of treatment and the females were allowed to litter and rear their progeny until Day 4 post-partum. At the end of the treatment period, Functional Observation Battery, motor activity and laboratory investigations (haematology and blood biochemistry) were carried out on five males and five females. The males were sacrificed after at least 5 weeks of treatment and the females on day 6 post-partum. Final body weights and selected organs weights (adrenals, brain, epididymides, heart, kidneys, liver, spleen, testes and thymus) were recorded and a complete macroscopic post-mortem examination was performed. A microscopic examination was performed on epididymis, gross lesions, mammary gland, ovary, prostate, seminal vesicle, testicle, uterus (incl. cervix and oviducts), vagina (all animals); adrenal gland, bone marrow (os femoris), brain (cerebrum, cerebellum, brain stem), heart (left and right ventricle, septum), intestine, small (duodenum, jejunum, ileum, incl. Peyer's patches, Swiss roll method), intestine, large (colon, rectum), kidney and ureter, liver, lungs (with mainstem bronchi and bronchioles), preserved by inflation with fixative and then immersion, lymph node (1 cervical, 1 mesenteric), nerve (sciatic), oesophagus, spinal cord (3 sections), spleen, stomach, thyroid (incl. parathyroids), thymus, regional lymph nodes, tongue (incl. base), trachea (incl. larynx), urinary bladder from five males and five females in the control and high dose groups.

There were no read across substance related deaths and related signs of toxicity were noted during the observational and neurological screenings. A reduction in food intake (-21.7%) during gestation/lactation period and body weight (-9.7%) during the laction period was observed in high dose females. No other read across substance related changes were noted for other parameters including, water consumption, haematology, clinical chemnistry, organ weights gross and histopathology. The effects observed at 1000 mg/kg bw/d in the females were considered adverse; thus, the NOAEL for systemic toxicity was determined to be 300 mg/kg bw/day (i.e., equivalent to 233.4 mg ai./kg bw/day) in females and 1000 mg/kg bw/day (i.e., equivalent to 778 mg a.i./kg bw/day) in male rats in this study (Hansen, 2013). Based on the results of the read across study, similar NOAELs can be considered for the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
NOAEL
233.4 mg/kg bw/day
Study duration:
subchronic
Species:
rat
Quality of whole database:
Guideline compliant studies on constituents
System:
other: no critical adverse effects were observed, except for reduction in food consumption and body weight in the study with isoalkylester constituent.

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - systemic effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Endpoint conclusion
Endpoint conclusion:
no study available

Repeated dose toxicity: dermal - local effects

Link to relevant study records
Reference
Endpoint:
short-term repeated dose toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Reason / purpose:
data waiving: supporting information
Endpoint conclusion
Endpoint conclusion:
no study available

Additional information

Oral:

In absence of repeated dose toxicity study with the test substance, the endpoint has been assessed based on studies for substances representative of the three main constituents, which can be categorised as phosphate esters (PSE), alcohol (hexadecanol) and isoalkylester (isostearyl isostearate). The results are presented below.

Constituent: PSE - read across study:

A screening study was conducted to determine the toxicity to reproduction of the read across substance, ‘mono- C12 PSE, Na+’ (purity: 100%, according to the OECD Guideline 422, in compliance with GLP. The read across substance was administered at 0 (control group), 250, 500 or 1000 mg/kg bw to male Sprague-Dawley SPF rats for 14 days before mating, through the mating period, and up to 1 d before necropsy (42 d in total) and to female Sprague-Dawley SPF rats for 14-d before mating, through the mating period and the gestation period, up to Day 4 of lactation (42 to 45 d in total) to investigate the repeated-dose, reproductive and developmental toxicities. In the 0 and 1000 mg/kg bw groups, a 14-d recovery period was allowed after the 42-d administration period to investigate the reversibility of the toxic changes. Administration of the read across substance had no effect on any of the following: general condition, findings on detailed observation of the general condition, results of function tests, grip strength, spontaneous motor activity, body weight, food consumption, results of urinalysis (including water intake), or in the results of haematological or blood chemistry tests. Administration of the read across substance had no effect on any of the following: general condition, findings on detailed observation of the general condition, results of function tests, grip strength, spontaneous motor activity, body weight, food consumption, results of urinalysis (including water intake), or in the results of haematological or blood chemistry tests. Changes in the forestomach may be considered to be adverse, however, they are also considered to be a result of local irritation of the (irritant) test substance (which is brought directly in contact to the mucosa in a massive amount by gavage application) than a true effect of systemic toxicity. Furthermore, in the light of the absence of a forestomach in humans, observed effects on this tissue are of questionable relevance with reference to the extrapolation of the toxic properties of a read across substance in humans. Under the study conditions, only (local) changes of the forestomach mucosa were observed at 250 mg/kg bw/day and above, which are due to the irritant properties of the substance. Therefore, the NOAEL (systemic) is considered to be 1000 mg/kg bw/day (BRC, 2005).

Constituent: Alcohol

A 28-d study was conducted to determine the sub-acute toxicity of hexadeacn-1-ol (purity: >95%) in Sprague-Dawley rats, according to a method similar to OECD Guideline 407, in compliance with GLP. In the study, SD rats (10 each sex/dose + 5 each sex/dose for reversibility) were exposed to the 0, 2, 10 and 20% test substance in olive oil, 5 mL/kg bw by oral gavage for 28 d, 5d/week. The actual doses received were calculated to be 0, 100, 500 and 1000 mg/kg bw/day respectively. Clinical signs, mortality and food consumption were observed daily, whereas, body weight and water consumption were analysed weekly. Opthalmoscopic examination was performed at the end of the study. Biochemistry and haematology analysis were performed 21/22 d after daily dosing. There were no treatment related effects on clinical signs, body weight, mortality and food consumption were noted. In haematology analysis, no differences between treated and control animals other than an increase in neutrophils containing rod like bodies observed in top dose females (1000 mg/kg bw/day) were noted. Statistically significant changes in some clinical chemical parameters were noted, where at 500 mg/kg/day, males increased potassium and females increased GGT, cholesterol and chloride, glucose was elevated in top dose males (1000 mg/kg/day). Both absolute and relative organ weights were essentially comparable in treated and control animals. Increases in absolute weight in male kidney at 500 mg/kg/day and male testes at 1000 mg/kg/day were noted. The only change in relative organ weight was an increase in male adrenal weight at 1000 mg/kg/day. Sporadic statistically significant changes in some organ weights and clinical chemical parameters were observed but these were not associated with histopathological changes and were not considered of toxicological significance. Under the study conditions, the NOAEL was considered to be >1000 mg/kg/day (i.e., >950 mg a.i./kg bw/day), based on lack of toxicologically significant treatment related effects at this dose level (top dose level) (OECD SIDS, 2006).

Constituent: Isoalkylester - read across study:

A study was conducted to determine the repeated dose toxicity of the read across substance, 'Isodecyl oleate' (purity: 77.8%), according to OECD Guideline 422, in compliance with GLP. Three groups of ten male and ten female Sprague-Dawley rats received the read across substance daily by oral (gavage) administration before mating, through mating and, for the females, through gestation until Day 3 post-partum. The read across substance was administered as a suspension in the vehicle, corn oil, at dose-levels of 0, 100, 300 or 1000 mg/kg bw/day. Another group of ten males and ten females received the vehicle alone, under the same experimental conditions and acted as a control group. The concentration of the dose formulation was checked start of treatment period; immediately after preparation of the read across substance-vehicle mixtures; 8 and 24 h after storage of the read across substance preparations at room temperature; end of treatment period; during treatment with the read across substance always before administration to the last animal of the dose level group.

The animals were checked at least twice daily during the dosing period for mortality and morbidity and once daily for clinical signs. Detailed clinical observations were performed once a week. Body weight and food consumption were recorded once a week during premating and mating periods (food consumption not during mating), and during gestation on post coital and lactation on Days 1 and 4 post-partum. The animals were paired for mating after 2 weeks of treatment and the females were allowed to litter and rear their progeny until Day 4 post-partum. At the end of the treatment period, Functional Observation Battery, motor activity and laboratory investigations (haematology and blood biochemistry) were carried out on five males and five females. The males were sacrificed after at least 5 weeks of treatment and the females on day 6 post-partum. Final body weights and selected organs weights (adrenals, brain, epididymides, heart, kidneys, liver, spleen, testes and thymus) were recorded and a complete macroscopic post-mortem examination was performed. A microscopic examination was performed on epididymis, gross lesions, mammary gland, ovary, prostate, seminal vesicle, testicle, uterus (incl. cervix and oviducts), vagina (all animals); adrenal gland, bone marrow (os femoris), brain (cerebrum, cerebellum, brain stem), heart (left and right ventricle, septum), intestine, small (duodenum, jejunum, ileum, incl. Peyer's patches, Swiss roll method), intestine, large (colon, rectum), kidney and ureter, liver, lungs (with mainstem bronchi and bronchioles), preserved by inflation with fixative and then immersion, lymph node (1 cervical, 1 mesenteric), nerve (sciatic), oesophagus, spinal cord (3 sections), spleen, stomach, thyroid (incl. parathyroids), thymus, regional lymph nodes, tongue (incl. base), trachea (incl. larynx), urinary bladder from five males and five females in the control and high dose groups.

There were no read across substance related deaths and related signs of toxicity were noted during the observational and neurological screenings. A reduction in food intake (-21.7%) during gestation/lactation period and body weight (-9.7%) during the laction period was observed in high dose females. No other read across substance related changes were noted for other parameters including, water consumption, haematology, clinical chemistry, organ weights gross and histopathology. The effects observed at 1000 mg/kg bw/d in the females were considered adverse; thus, the NOAEL for systemic toxicity was determined to be 300 mg/kg bw/day (i.e., equivalent to 233.4 mg ai./kg bw/day) in females and 1000 mg/kg bw/day (i.e., equivalent to 778 mg a.i./kg bw/day) in male rats in this study (Hansen, 2013). Based on the results of the read across study, similar NOAELs can be considered for the test substance,'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'.

Overall, based on the available weight of evidence, except for reduction in food consumption and body weight changes, no other toxicologically significant or critical systemic effects are expected for the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate'. However, as a conservative approach, the lowest NOAEL value of 300 mg/kg bw/day (or 233.4 mg a.i./kg bw/day; based on read across to isoalkylester study) has been considered further for hazard/risk assessment.

Dermal:

A repeated dose dermal toxicity study is not required because an OECD 422 and 28-day oral studies are available for substances representative of the main constituents. Further, given the physico-chemical properties of the substance, dermal absorption is not expected to be higher than via the oral route. Hence, testing via dermal route will less likely result in any additional hazard identification and further testing involving vertebrate animals may be omitted, in accordance with Annex XI (1.2) of the REACH regulation. Nevertheless, the risk assessment for this route has been carried out based on oral studies available with the read across substances, using appropriate route-to-route extrapolation assessment factors as per the ECHA Guidance R.8.

Inhalation:

The substance is a solid (white pellets) with a low vapour pressure at room temperature. Due to its physical state and physical chemical properties it is unlikely that this substance will form inhalable dust, mist or fumes during normal processing and use conditions. In case inhalable forms of the substance are created under particular conditions (e. g. spraying, elevated temperature/pressure), appropriate risk management measures such as closed systems, exhaust ventilation or wearing of respirators are implemented to control exposure. Under such conditions, the risk to humans following inhalation exposure can be considered minimal and further testing involving vertebrate animals may be omitted, in accordance with Annex XI (1.2) of the REACH regulation. Nevertheless, the risk assessment for this route has been carried out based on oral studies available on substances representative of the main constituents, using appropriate route-to-route extrapolation assessment factors as per the ECHA Guidance R.8.

Justification for classification or non-classification

Based on the available weight of evidence from the studies on the main constituents, the test substance, 'mono- and di- C16 PSE, K+ and C16-OH and isostearyl isostearate', does not warrant a classification for STOT RE, according to the EU CLP criteria (Regulation 1272/2008/EC).