Alcohols, C11-15-secondary, ethoxylated

Repeated dose toxicity: oral

Administrative data

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26 August 2015 to 31 March 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 408 (Repeated Dose 90-Day Oral Toxicity Study in Rodents)

Version / remarks:

September 21, 1998

Deviations:

no

GLP compliance:

yes

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

Alcohols, secondary C11-15, ethoxylated

EC Number:

614-295-4

Cas Number:

68131-40-8

Molecular formula:

C(11-15) H(23-31) O (C2H4O)xH where n= approximately 3

IUPAC Name:

Alcohols, secondary C11-15, ethoxylated

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Sponsor; Lot No. 15E25
- Expiration date of the lot/batch: Not reported
- Purity test date: Not reported
STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Room temperature, protected from light and airtight
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: soluble in corn oil
- Reactivity of the test substance with the solvent/vehicle of the cell culture medium: None
TREATMENT OF TEST MATERIAL PRIOR TO TESTING
- Treatment of test material prior to testing: none
- Preliminary purification step (if any): N/A
- Final dilution of a dissolved solid, stock liquid or gel: final solution in corn oil

Test animals

Species:

rat

Strain:

other: Crl:CD(SD) SPF

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: Charles River Laboratories, Japan Inc
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 6 weeks
- Weight at study initiation: Males: 210-247g; Females: 143-187g
- Fasting period before study: none
- Housing: individually during study
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum): ad libitum
- Acclimation period: 6 days
ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24
- Humidity (%): 39-54
- Air changes (per hr): 10-15
- Photoperiod (hrs dark / hrs light): 12/12
IN-LIFE DATES: From: 9 September 2015 To: 23 December 2015

Administration / exposure

Route of administration:

oral: gavage

Details on route of administration:

Dosing solutions were administered into the stomach by oral gavage using a disposable gastric tube and
a disposable syringe.

Vehicle:

corn oil

Details on oral exposure:

- Justification for use and choice of vehicle (if other than water): poor solubility in water
- Concentration in vehicle: 0, 20, 60, 200 mg/mL
- Amount of vehicle (if gavage): 5 mL/kg bodyweight
- Lot/batch no. (if required): V5G5873
- Purity: Not reported

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Analysis of content of dosing solutions was by LC/MS/MS and was carried out at the start of the study (7
Sept 2015), at an intermediate time point (14 Oct 2015) and at the end of the study (4 Dec 2015). (See
Table 1 below). Stability of the test material in corn oil was determined prior to the start of the study d
uring storage in the refrigerator for 7 days followed by 24 h at room temperature.

Duration of treatment / exposure:

90 Days

Frequency of treatment:

Once Daily

Doses / concentrations

Doses / concentrations 1

Dose / conc.:

0 mg/kg bw/day (nominal)

Doses / concentrations 2

Dose / conc.:

100 mg/kg bw/day (nominal)

Remarks:

Individual dose volume was calculated based on the body weight on the measurement day closest to each day of administration.

Doses / concentrations 3

Dose / conc.:

300 mg/kg bw/day (nominal)

Remarks:

Individual dose volume was calculated based on the body weight on the measurement day closest to each day of administration.

Doses / concentrations 4

Dose / conc.:

1 000 mg/kg bw/day (nominal)

Remarks:

Individual dose volume was calculated based on the body weight on the measurement day closest to each day of administration.

No. of animals per sex per dose:

10 in toxicity test groups
5 in recovery groups

Details on study design:

- Dose selection rationale: In the preliminary study of Alcohols, C11-15-secondary, ethoxylated in rats
(SR15064; dose levels 100, 300, and 1000 mg/kg), no clear effects of the test article administration were
noted, except an increase in the liver weight in the high dose of 1000 mg/kg group. Based on the result,
for the 90-day repeated dose oral toxicity study, 1000 mg/kg that is the maximum feasible dose, was
selected as the high dose level, and 300 and 100 mg/kg were used by dividing the high dose level by a
common ratio of approximately 3
- Rationale for animal assignment (if not random): Animals were assigned to test groups by the stratified
random sampling method using the toxicological data processing system (MiTOX, Mitsui Zosen Systems
Research Inc.), so that group mean body weight on the day of group assignment (the day before the first
administration) was comparable.
- Post-exposure recovery period in satellite groups: S-week recovery period for highest concentraton
treatment group and a control (5 animals per sex per group)

Examinations

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: Twide daily, before and after administration; twice daily during recovery period (am and
pm)
- Cage side observations were included. - Animals were observed for mortality, external appearance,
behavior and other changes. Any abnormalities noted were recorded with the times of onset or discovery.
DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: Once before the start of administration and on Administration Day 7, and once a week
thereafter (including the recovery period)
BODY WEIGHT: Yes
- Time schedule for examinations: During administration period: on Administration Days 1, 4,
and 7, and every 7 days thereafter; before dosing on each measurement day, and the day of necropsy.
During recovery period: on Recovery Days 1 and 7, and every 7 days thereafter.
FOOD CONSUMPTION AND COMPOUND INTAKE (if feeding study):
During administration period: on Administration Days 1, 4, and 7, and every 7 days thereafter
During recovery period: on Recovery Days 1 and 7, and every 7 days thereafter
OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: Before the start of administration (during acclimatization
period), and during Administration Week 13 and Recovery Week 2.
- Dose groups that were examined: control and high dose groups
HAEMATOLOGY: Yes
- Time schedule for collection of blood: at necropsy
- Anaesthetic used for blood collection: Yes, IP injeciton of sodium pentobarbital
- Animals fasted: Yes
- How many animals: All
- Parameters checked in table No.2 were examined.
CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: at necropsy
- Animals fasted: Yes
- How many animals: All
- Parameters checked in table No.3 were examined.
URINALYSIS: Yes
- Time schedule for collection of urine: Administration Week 13 and Recovery Week 2
- Metabolism cages used for collection of urine: Yes
- Animals fasted: No
- Parameters checked in table No.4 were examined.
NEUROBEHAVIOURAL EXAMINATION: Yes
- Time schedule for examinations: Administration Week 13 and Recovery Week 2
- Dose groups that were examined: All
- Battery of functions tested: sensory activity / grip strength / motor activity
IMMUNOLOGY: No

Sacrifice and pathology:

GROSS PATHOLOGY: Yes (see table)
HISTOPATHOLOGY: Yes (see table)

Results and discussion

Results of examinations

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
No significant difference was noted in body weight in males or females in any test article group compared to the control group. Body weight gain on Administration Days 4 and 7 was significantly low in males in the 1000 mg/kg group. Body weight gain of females in the 300 mg/kg group was significantly low on
Administration Day 4. In females in the high dose of 1000 mg/kg group, however, there were no significant differences. These results indicated that the low body weight gain was unrelated to the test article administration. In males in the 300 mg/kg group and males and females in the 100 mg/kg group, no significant differences were noted in the body weight gain compared to the control group.

[Recovery period]
No significant difference was noted in body weight in males or females in any test article group compared to the control group. Body weight gain on Recovery Day 14 was significantly high in males in the 1000 mg/kg group. This was considered incidental and unrelated to the test article administration because no
inhibitory trend was noted in body weight, other than the early stage of administration period, and no increasing trend was shown in body weight thereafter.
In body weight gains, no significant difference was noted in females in the 1000 mg/kg group compared to the control group.

Food consumption and compound intake (if feeding study):

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
Food consumption was significantly low in males and females in the 1000 mg/kg group on Administration Day 4, which recovered after that. Food consumption was significantly high in the 1000 mg/kg group on dosing Day 90 in males, and on Days 84 to 90 in females.
These were considered unrelated to the test article administration because they were within the range of historical control data of the test facility (12.3 to 34.7 g, 25.18 ± 3.84 g in males on Day 90; 12.3 to 26.8 g, 19.50 ± 3.33 g in females on Day 84, and 10.1 to 27.5 g, 15.99 ± 2.67 g in females on Day 90).
No significant differences were noted in food consumption in males or females in the 100 or 300 mg/kg group, compared to the control group.

[Recovery period]
Food consumption was significantly high in males in the 1000 mg/kg group on Recovery Days 7 and 14,which were within the range of historical control data of the test facility (21.9 to 39.7 g, 30.12 ± 4.95 g on Recovery Day 7; 21.8 to 39.3 g, 29.50 ± 5.13 g on Recovery Day 14). These were also considered incidental and unrelated to the test article administration.

Food efficiency:

not examined

Water consumption and compound intake (if drinking water study):

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
No significant differences were noted in any parameter in males or females in any test article group compared to the control group.

[Recovery period]
In females in the 1000 mg/kg group, RBC was significantly high, while there were no increasing trends in HGB or HCT, and RBC was within the range of historical control data of the test facility (RBC: 667 to 949 × 104/μL, 835.5 ± 42.2 × 104/μL).
This change was not noted in the examination at the end of administration period; therefore, this change was considered unrelated to the test article administration. No significant differences were noted in any other parameters in males or females compared to the control group.

Clinical biochemistry findings:

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
γ-GTP was significantly high in males in the 1000 mg/kg group; however, no changes indicating effects on the liver function were noted in the other parameters, and histopathological examination of the liver revealed no toxic changes. In addition, these values were within the range of historical control data of the test facility (γ-GTP 0.0 to 2.4 IU/L, 0.45 ± 0.30 IU/L) 2). This change was therefore considered not indicative of toxic changes attributable to the test article administration.
T-Cho was significantly high in males and females in the 1000 mg/kg group. As these values were within the range of historical control data of the test facility (33 to 109 mg/dL and 58.7 ± 13.7 mg/dL in males and 32 to 112 mg/dL and 67.5 ± 13.6 mg/dL in females), this change was considered unrelated to the test
article administration.
AST were significantly low in males, and T-Bil was significantly low in males and females in the 1000 mg/kg group. These were within the range of historical control data of the test facility (Males: AST 40 to 147IU/L, 63.2 ±14.4 IU/L, T-Bil 0.03 to 0.12 mg/dL, 0.063 ± 0.014 mg/dL. Females: T-Bil 0.04 to 0.18 mg/dL,
0.076 ± 0.020 mg/dL) and were low values; therefore, these were considered unrelated to the test article administration.
α1-G was significantly high in females in the 100, 300, and 1000 mg/kg groups. These were not observed in a dose-related manner, and there were no differences in A/G ratio, albumin, or other protein fractions. In addition, α1-G was within the range of historical control data of the test facility (9.9 to 20.5%, 16.72 ±
1.70%). The high α1-G was therefore considered unrelated to the test article administration.
No significant differences were noted in any other parameter in males or females in any test article group compared to the control group.

[Recovery period]
Ca was significantly high in males in the 1000 mg/kg group. No significant difference was noted at the end of administration period and Ca was within the range of historical control data of the test facility (8.9 to 10.5 mg/dL, 9.71 ± 0.33 mg/dL); therefore, this was considered unrelated to the test article administratio
n. In males in this group, albumin (calculated from TP and protein fractions) was significantly high, which was considered unrelated to the test substance because there was no difference in albumin of protein fraction, and this change was not noted in the examination at the end of administration period.
No significant differences were noted in any other parameter in males or females in any test article group compared to the control group.

Urinalysis findings:

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
In males in the 1000 mg/kg group, pH was significantly low, which was considered unrelated to the test article administration because they were within the range of historical control data of the test facility (pH 6.5 to 8.5).
In females in the 300 mg/kg group, specific gravity was significantly low, which was considered unrelated to the test article administration because this was not noted in the 1000 mg/kg group.

[Recovery period]
No significant differences were noted in any parameter in males or females in any test article group compared to the control group.

Behaviour (functional findings):

effects observed, non-treatment-related

Description (incidence and severity):

[Administration period]
The motor activity counts were significantly high at 10’ – 20’, 30’ – 40’, 40’ – 50’, and 0’ – 60’ after the start of measurement in females in the 300 mg/kg group; and at 10’ – 20’, 20’ – 30’, and 0’ – 60’ after the start of measurement in females in the 1000 mg/kg group. These changes were not observed in a dose-related manner in the 300 or 1000 mg/kg group, and were within the range of historical control data of the test facility (10’ – 20’ after the start of measurement, 199 to 1593, 676.2 ± 267.8; 20’ – 30’, 81 to 1379, 511.3 ± 254.7; 30’ – 40’, 4 to 1051, 396.6 ± 227.5; 40’ – 50’, 0 to 1338, 323.7 ± 239.4; 0’ – 60’, 929 to 7800, 2983.1 ± 1286.2) 2). These changes were therefore considered unrelated to the test article administration. No significant differences were noted in any other parameters in males or females compared to the control group.

[Recovery period]
No significant differences were noted in any parameters in males or females compared to the control group.

Immunological findings:

not examined

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

[Administration period]
The weight relative to body weight (relative weight) of the liver and kidney was significantly high in males in the 1000 mg/kg group.The absolute and relative weight of the liver and kidney was significantly high in females in the 300 and 1000 mg/kg groups. In females, absolute kidney weight was significantly high also in the 100 mg/kg group. The liver weight of females in the 300 mg/kg group and kidney weight of females in the 100 to 1000 mg/kg groups were within the range of historical control data of the test facility (liver, 5.49 to10.91 g, 7.319 ± 0.915 g; kidney, 1.51 to 2.95 g, 1.974 ± 0.231 g).
The absolute and relative thymus weight was significantly high in males in the 300 mg/kg group. This changes was not noted in the high dose of 1000 mg/kg group, and the values were within the range of historical control data of the test facility (70 to 645 mg/dL, 314.3 ± 84.6 mg/dL)2); therefore, these were c
onsidered unrelated to the test article administration.

[Recovery period]
In females in the 1000 mg/kg group, relative adrenal weight was significantly high. There was no significant difference at the end of administration period, and the absolute adrenal weight was within the range of historical control data of the test facility (49 to 106 mg, 72.3 ± 11.2 mg); therefore, this was considered
unrelated to the test article administration.

Gross pathological findings:

effects observed, treatment-related

Description (incidence and severity):

[Administration period]
In the 1000 mg/kg group, thickening of the forestomach mucosa was noted in 3 males and 1 female, and focal thickening of the forestomach mucosa in 1 male and 4 females.
In the 300 mg/kg and lower dose groups, no abnormal findings were noted other than grayish white subcutaneous mass noted in 1 female in the control group.

[Recovery period]
No abnormal findings were noted in any animal.

Neuropathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

[Administration period]
In the 1000 mg/kg group, squamous cell hyperplasia of the forestomach was noted in 4 males (slight: degree is shown in parentheses hereinafter), 2 males (mild), 1 male (moderate), 3 females (slight), and 2 females (mild); ulcer of the forestomach in 2 males (slight) and 1 female (mild); infiltration of inflammatory
cells in the stomach submucosa in 3 males (slight), 1 male (mild), and 2 females (slight); edema of the stomach submucosa in 2 males (slight), 1 male (mild), 2 females (slight), and 1 females (mild). Hemorrhage of stomach submucosa was noted in 1 male (slight) in the 1000 mg/kg group.
Squamous cell hyperplasia of the forestomach was noted in 3 males (slight) in the 300 mg/kg group.
Centrilobular hypertrophy of hepatocytes in the liver was noted in 5 males and 5 females (slight) in the 1000 mg/kg group.
Hypertrophy of follicular cells in the thyroid was noted in 4 males (slight) in the 1000 mg/kg group.
In the kidneys, eosinophilic body of proximal tubular epithelium was noted in 2 males (slight) in the 100 mg/kg group, 4 males (slight) in the 300 mg/kg group, 4 males (slight) and 3 males (mild) in the 1000 mg/kg group. As the microscopic specimens were positive for α2μ-globulin antibody, this change was likely to
be α2μ-globulin nephropathy that is specific to male rats, and was noted also in 2 males (slight) in the control group. These indicated increased severity of eosinophilic body of proximal tubular epithelium in the 1000 mg/kg group.
Other than those described above, abnormalities were noted in the heart, pancreas, pituitary gland, eye balls, liver, kidneys, prostate, and ovaries. These were considered unrelated to the test article administration because they were noted also in the control group, not observed in a dose-related manner, or o
bserved in only a few animals (1 or2).

[Recovery group]
Squamous cell hyperplasia of the forestomach was noted in 2 males (slight) in the 1000 mg/kg group, which was not noted in females.
In the liver, microgranuloma was noted in 3 males and 3 females (slight) in the 1000 mg/kg group; however, centrilobular hypertrophy of hepatocytes was not noted.
Hypertrophy of follicular cells was noted in the thyroid of 2 males (slight) in the 1000 mg/kg group.
In the kidneys, eosinophilic body in the proximal tubular epithelium was noted in 1 male (slight) in the control group, and in 2 males (slight) in the 1000 mg/kg group.
All these changes were recovered or tended to recover.

Histopathological findings: neoplastic:

not examined

Other effects:

no effects observed

Effect levels

1

Key result

Dose descriptor:

NOAEL

Effect level:

<= 300 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

female

Basis for effect level:

other: irritation of the stomach mucosa

2

Key result

Dose descriptor:

NOAEL

Effect level:

<= 100 mg/kg bw/day (nominal)

Based on:

test mat.

Sex:

male

Basis for effect level:

other: irritation of the stomach mucosa

Target system / organ toxicity

Critical effects observed:

no

Applicant's summary and conclusion

Conclusions:

Softanol 30 was administered to rats by oral gavage for 90 days at doses of 0, 100, 300 amnd 1000 mg/kg/bw/day. Irritation of the stomach mucosa was noted at 1000 mg/kg in females and 300 mg/kg and higher doses in males; therefore, the no-observed-adverse-effect level (NOAEL) of Alcohols, C11-15-secondary, ethoxylated was considered to be 100 mg/kg in males and 300 mg/kg in females.

Executive summary:

Alcohols, C11-15-secondary, ethoxylated was orally administered to male and female rats for 90 days to investigate its potential to induce toxicity and its profile. Following the end of administration, recovery from toxic effects was assessed during a 14-day withdrawal period. In the 1000 mg/kg group, transient inhibition of body weight gain or food consumption was noted at the start of administration in males and females. In the 1000 mg/kg group, necropsy and histopathological examination revealed abnormalities in the forestomach mucosa in both males and females. These changes were likely due to the irritancy of the test substance; however, they were not noted in the necropsy of the preliminary study of Alcohols, C11-15- secondary, ethoxylated (SR15064). Accordingly, these changes were considered influenced by the duration of administration. Histopathological changes in the forestomach mucosa, which were slight in degree, were noted also in males in the 300 mg/kg group. As the severity of the changes in the forestomach mucosa was reduced during the recovery period, they were considered reversible. The liver weight was high and centrilobular hypertrophy of hepatocytes was noted in both males or females in the 1000 mg/kg group. In addition, hypertrophy of follicular cells in the thyroid was noted in males in this group. These changes were considered hepatic enzyme induction, which are adaptive changes accompanying drug-induced metabolic activation of the liver, and their secondary changes. The changes in the liver, of which severity was reduced during the recovery period, were considered to be reversible. The increased liver weight in females in the 300 mg/kg group was considered unrelated to the test article administration because it was within the range of historical control data of the test facility, and no histological changes indicated hepatic enzyme induction. In males in the 1000 mg/kg group, eosinophilic body of proximal tubular epithelium, which was likely to be α2μ-globulin nephropathy that is specific to male rats, was increased in severity compared to that in the other groups. In addition, kidney weight was increased in males of this group. The renal changes were considered to be reversible because their severity was reduced during the recovery period. In females, kidney weight was high in the 100 to 1000 mg/kg groups, which was considered unrelated to the test article administration because it was within the range of historical control data of the test facility and no related histopathological changes were detected. As described above, irritation of the stomach mucosa was noted at 1000 mg/kg in females and 300 mg/kg and higher doses in males; therefore, the no-observed-adverseeffect level (NOAEL) of Alcohols, C11-15-secondary, ethoxylated was considered to be 100 mg/kg in males and 300 mg/kg in females.