Tricyclodecanedimethanol

Endpoint summary

Description of key information

The subchronic NOAEL was 1000 mg/kg bw/day in an OECD 408 guideline study using male and female rats,  and there were no adverse effects noted on the male and female reproductive organs, oestrous cycle, and sperm parameters. This is in line with the results of a screening study. In a combined oral repeated dose toxicity study (28 d) with a reproduction/developmental toxicity screening test under GLP conditions (OECD TG 422), the subacute parental NOAEL of octahydro-4,7-methano-1H-indenedimethanol (TCD Alcohol DM) was 600 mg/kg bw/day.
The vapour pressure of TCD-Alcohol DM is determined to be < 1 hPa (sect. 4.6). Atmosphere concentrations attainable under ambient conditions are assessed to be too low to exert toxic effects.

Key value for chemical safety assessment

Repeated dose toxicity: via oral route - systemic effects

Link to relevant study records

Reference

Endpoint:

sub-chronic toxicity: oral

Type of information:

experimental study

Adequacy of study:

key study

Study period:

01 Feb 2013- 25 Oct 2013

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted to OECD guidelines and in compiance with GLP

Qualifier:

according to guideline

Guideline:

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

Deviations:

yes

Remarks:

Food consumption was recoreded for pre-treatment week. Method of analysis changed from HPLC to LC-MS/MS and LOD/LOQ not assessed. Any response form blank samples must be <20% lowest calibration standard.

GLP compliance:

yes (incl. QA statement)

Remarks:

Certificate of compliance issued by UK MHRA included in report

Limit test:

no

Species:

rat

Strain:

Wistar

Sex:

male/female

Details on test animals or test system and environmental conditions:

TEST ANIMALS
- Source: UK
- Age at study initiation: 41-47 days
- Weight at study initiation: Males = 133-173g: Females = 118-150g
- Housing: Cage: Polycarbonate body with a stainless steel mesh lid, changed at
appropriate intervals. Bedding: Wood based bedding which was changed at appropriate
intervals each week.
- Diet (e.g. ad libitum): Rat and Mouse No. 1 Maintenance Diet.
- Water (e.g. ad libitum): Potable water from the public supply. Non-restricted via polycarbonate bottles with sipper tubes.
- Acclimation period:12 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19-23 °C
- Humidity (%):40-70 %
-Air supply: Filtered fresh air which was passed to atmosphere and not recirculated.
- Photoperiod (hrs dark / hrs light): 12 h dark/12 hlight

IN-LIFE DATES: From: 07 May 2013 To: 28 Oct 2013

Route of administration:

oral: gavage

Vehicle:

propylene glycol

Remarks:

+ ethyl acetate (w.w)

Details on oral exposure:

PREPARATION OF DOSING SOLUTIONS:

VEHICLE
- Concentration in vehicle:0, 50, 100, 200 mg/ml
- Amount of vehicle (if gavage): 5 ml/kg

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

Homogeneity and stability assessed before commencement of treatment and test material found to be stable for >24h at 2-8 deg C and for 15 days at ambient temperature.

Duration of treatment / exposure:

13 weeks

Frequency of treatment:

Daily

No. of animals per sex per dose:

10 males and 10 females per treatment group

Control animals:

yes, concurrent vehicle

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: twice a day

DETAILED CLINICAL OBSERVATIONS: Yes
- Time schedule: twice daily

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

OPHTHALMOSCOPIC EXAMINATION: Yes
- Time schedule for examinations: per-treatment and week 11

HAEMATOLOGY: Yes
- Time schedule for collection of blood: week 13
- Anaesthetic used for blood collection: Yes (isoflurane)
- Animals fasted: Yes
- How many animals: all
- Parameters checked: according to OECD 408

CLINICAL CHEMISTRY: Yes
- Time schedule for collection of blood: week 13
- Animals fasted: Yes
- How many animals: all
- Parameters checked: according to OECD 408

NEUROBEHAVIOURAL EXAMINATION: Yes
- Sensory reactivity and grip strength
- Touch response
- Huntingdon Life Sciences: PJJ0003
- Auditory startle reflex
- Tail pinch response
- Grip strength
- Motor activity

OTHER:
Sperm analysis: Immediately after scheduled sacrifice of each male, the left vas deferens, epididymis and testis was removed and the epididymis and testis were weighed.
-Analysis of: sperm motility, sperm morphology, sperm count

Oestrous cycles - vaginal smears

Sacrifice and pathology:

GROSS PATHOLOGY: Yes
HISTOPATHOLOGY: Yes

Statistics:

All statistical analyses were carried out separately for males and females.
The following data types were analysed at each timepoint separately:
Body weight, using gains over appropriate study periods
Grip strength and motor activity
Haematology and blood chemistry
Oestrous cycles
Organ weights, absolute and adjusted for terminal body weight

Clinical signs:

effects observed, treatment-related

Description (incidence and severity):

There were three deaths during treatment. Female Nos. 132, 136 and 140 receiving 1000 mg/kg/day were despatched for welfare reasons on Day 12, 80 and 13 of treatment respectively. Clinical signs prior to despatch included; Female No. 132: decreased activity, laboured/shallow/slow breathing, râles (wet), sneezing and dark eyes. Female No. 136: excessive chewing, irregular/laboured breathing, râles (wet), piloerection, and prominent eyes. Female No. 140: decreased activity, laboured/shallow breathing, râles (wet), piloerection, elevated gait and distended abdominal area. Macroscopic examination revealed abnormal contents (gas) and/or distension of the caecum, colon, ileum, jejunum, stomach and/or incomplete lung collapse in each of these animals with no direct histopathologic correlate. In addition, histopathological examination revealed: No. 132, minimal haemorrhage that correlated to dark, multilobular, diffuse areas of the lungs and minimal apoptosis of the thymus correlating to dark thymus areas; No. 136, small spleen within no microscopic correlate, minimal lung and bronchi perivascular cell infiltrate and inflammation, minimal apoptosis of the thymus correlating to dark thymus areas; No. 140, minimal apoptosis and decreased cellularity of the thymus with no macroscopic correlate and a small spleen with slight white pulp atrophy observed microscopically.

Mortality:

mortality observed, treatment-related

Description (incidence):

There were three deaths during treatment. Female Nos. 132, 136 and 140 receiving 1000 mg/kg/day were despatched for welfare reasons on Day 12, 80 and 13 of treatment respectively. Clinical signs prior to despatch included; Female No. 132: decreased activity, laboured/shallow/slow breathing, râles (wet), sneezing and dark eyes. Female No. 136: excessive chewing, irregular/laboured breathing, râles (wet), piloerection, and prominent eyes. Female No. 140: decreased activity, laboured/shallow breathing, râles (wet), piloerection, elevated gait and distended abdominal area. Macroscopic examination revealed abnormal contents (gas) and/or distension of the caecum, colon, ileum, jejunum, stomach and/or incomplete lung collapse in each of these animals with no direct histopathologic correlate. In addition, histopathological examination revealed: No. 132, minimal haemorrhage that correlated to dark, multilobular, diffuse areas of the lungs and minimal apoptosis of the thymus correlating to dark thymus areas; No. 136, small spleen within no microscopic correlate, minimal lung and bronchi perivascular cell infiltrate and inflammation, minimal apoptosis of the thymus correlating to dark thymus areas; No. 140, minimal apoptosis and decreased cellularity of the thymus with no macroscopic correlate and a small spleen with slight white pulp atrophy observed microscopically.

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not examined

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

not examined

Ophthalmological findings:

no effects observed

Haematological findings:

effects observed, treatment-related

Description (incidence and severity):

Week 13 : when compared with the controls, a statistically significant reduction in activated partial thromboplastin time in males which was dose-related and females receiving 500 or 1000 mg/kg/day (not statistically significant)

Clinical biochemistry findings:

no effects observed

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

effects observed, treatment-related

Description (incidence and severity):

high absolute and body weight-adjusted liver weights in males and females at 500 or 1000 mg/kg/day; high body weight-adjusted kidney weight in males with a dose-relationship and females given 1000 mg/kg/day apparent

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Changes seen in nasal turbinates related to teatment

Histopathological findings: neoplastic:

effects observed, treatment-related

Description (incidence and severity):

Changes related to treatment with TCD Alcohol DM were seen in the nose/turbinates.

Details on results:

Findings comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx, degeneration/regeneration/inflammatory cell infiltration of the epithelium and deformity/fusion of turbinates. They were considered treatment related in all groups of both sexes.
Minimal findings, although not including turbinate changes, were seen in a single control female.
Changes of an uncertain relationship to treatment with TCD Alcohol DM were seen in the stomach.
Stomach - Epithelial hyperplasia of the non-glandular stomach was seen in occasional animals of both sexes given 500 and 1000 mg/kg/day and a single male given 250 mg/kg/day. In addition, a single case of erosion was seen in the non-glandular region of the stomach of a female given
1000 mg/kg/day. The low numbers affected and lack of consistency between the two sexes made this finding have an uncertain relationship to treatment.
All other microscopic findings were considered to be incidental and unrelated to treatment with TCD Alcohol DM.

No effects on sperm motility, morphology or concentration were observed after treatment at doses up to 1000 mg/kg/day.

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: highest dose tested

Critical effects observed:

not specified

There were three deaths during treatment. Female Nos. 132, 136 and 140 receiving 1000 mg/kg/day were despatched for welfare reasons on Day 12, 80 and 13 of treatment respectively.

Clinical signs prior to despatch included;

Female No. 132: decreased activity, laboured/shallow/slow breathing, râles (wet), sneezing and dark eyes.

Female No. 136: excessive chewing, irregular/laboured breathing, râles (wet), piloerection, and prominent eyes.

Female No. 140: decreased activity, laboured/shallow breathing, râles (wet), piloerection, elevated gait and distended abdominal area.

Macroscopic examination revealed abnormal contents (gas) and/or distension of the caecum, colon, ileum, jejunum, stomach and/or incomplete lung collapse in each of these animals with no direct histopathologic correlate.

In addition, histopathological examination revealed:

No. 132, minimal haemorrhage that correlated to dark, multilobular, diffuse areas of the lungs and minimal apoptosis of the thymus correlating to dark thymus areas;

No. 136, small spleen within no microscopic correlate, minimal lung and bronchi perivascular cell infiltrate and inflammation, minimal apoptosis of the thymus correlating to dark thymus areas;

No. 140, minimal apoptosis and decreased cellularity of the thymus with no macroscopic correlate and a small spleen with slight white pulp atrophy observed microscopically.

Microscopic findings were seen in the nose and comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx, degeneration/regeneration/inflammatory cell infiltration of the epithelium and, deformity/fusion of turbinates. The microscopic changes in the nose/turbinates in two of the decedents (No. 132, 140) were of a severity to conclude that they were the factor responsible for the death whereas in the remaining animal (No. 136) these findings were not of a severity to be certain as to the cause of death. Similar findings in the nose/turbinates were also seen in terminal animals.

Conclusions:

It was concluded, that for the assessment of systemic toxicity, oral administration of octahydro-4,7-methano-1H-indenedimethanol (TCD Alcohol DM), an industrial chemical intermediate, to RccHan™;WIST (Han Wistar) rats at dose levels of 250, 500 and 1000 mg/kg/day for 13 weeks the no-observed-adverse-effectlevel (NOAEL) in this study was considered to be 1000 mg/kg/day, and there were no adverse effects noted on the male and female reproduction organs, oestrous cycle, and sperm parameters.

It is acknowledged that adverse treatment related changes with TCD Alcohol DM were seen in the nose/turbinates of all treated groups of both sexes. Nasal findings comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx,
degeneration/regeneration/inflammatory cell infiltration of the epithelium and deformity/fusion of turbinates. This pathology was considered to
be secondary to retrograde aspiration of material from the oropharynx into the nasopharynx and nasal cavity, this is considered to be a local effect, associated with the route of administration of this formulation and as such has been excluded from the assessment of systemic toxicity.

Executive summary:

The systemic toxic potential of TCD Alcohol DM, to Han Wistar (RccHanTM:WIST) rats by oral gavage administration was assessed over a period of 13 weeks. Three groups, each comprising ten males and ten females, received TCD Alcohol DM at doses of 250, 500 or 1000 mg/kg/day. A similarly constituted control group received the vehicle, 16% (w/w) ethylacetate in propylene glycol, at the same volume dose as the treated groups.

During the study, clinical condition, detailed physical and arena observations, sensory reactivity, grip strength, motor activity, body weight, food consumption, ophthalmoscopy, haematology (peripheral blood), blood chemistry, oestrous cycles, sperm analysis, organ weight, macropathology and histopathology investigations were undertaken.

Results:

Three females receiving 1000 mg/kg/day were killed for welfare reasons an say 12, 13 and 80 due to marked breathing impairment. Clinical signs prior to despatch included laboured/ shallow/irregular/slow breathing, räles (wet), decreased activity, piloerection, elevated gait, excessive chewing and distended abdomen. Macroscopic examination revealed abnormal contents (gas) and/or distension of the caecum, colon, ileum, jejunum, stomach and/or incomplete lung collapse in each of these animals with no direct histopathologic findings but associated with the histopathological changes in the nasal turbinates detailed in the following paragraph. The following section includes additional macropathology and histopathological examination findings: No. 132, minimal haemorrhage that correlated to dark, multilobular, diffuse areas of the Jungs and minimal apoptosis of the thymus correlating to dark thymus areas; No. 136, small spieen within no microscopic correlate, minimal lung and bronchi perivascular cell infiltrate and inflammation, minimal apoptosis of the thymus correlating to dark thymus areas; No. 140, minimal apoptosis and decreased cellularity of the thymus with no macroscopic correlate and a small spieen with slight white pulp atrophy observed microscopically.

Microscopic findings were seen in the nose and comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx, degeneration/regeneration/inflammatory cell infiltration of the epithelium and, deformity/fusion of turbinates. The microscopic changes in the nose/turbinates in two of the decedents (No. 132, 140) were of a severity to conclude that they were the factor responsible for the death whereas in the remaining animal (No. 136) these findings were not of a severity to be certain as to the cause of death. Similar findings in the nose/turbinates were also seen in terminal animals.

Signs associated with dosing included räles and rapid breathing in individual animals at 500 or 1000 mg/kg/day during treatment. It is considered that this sign is predominately related to the dosing procedure and potential reflux of the formulation, rather than a systemic effect of treatment. Similar signs have been observed at physical examination in a few treated animals.

There were no treatment-related findings at the sensory reactivity, grip strength or motor activity investigations. Oestrous cycles were also considered unaffected by treatment.

 Body weight change and food consumption was unaffected by treatment with TCD Alcohol DM.

There were no treatment-related ophthalmoscopic findings.

During Week 13 of treatment haematological investigations revealed a shortening of activated partial thromboplastin times in males receiving 250, 500 or 1000 mg/kg/day and to a lesser degree in females receiving 500 or 1000 mg/kg/day. In addition, blood chemistry examination revealed slightly high alkaline phosphatase activities in males receiving 1000 mg/kg/day, although in the absence of any change in associated parameters (ALT and AST), the significance of this is unclear and also slightly high cholesterol concentration in these males.

Organ weight changes that were considered to be related to treatment were high absolute and body weight-adjusted liver weights in males and females at 500 or 1000 mg/kg/day, and high body weight-adjusted kidney weight in males with a dose-relationship apparent (absolute kidney weights were also slightly high) and females given 1000 mg/kg/day. In addition, there were high body weight-adjusted spieen weights in males given 1000 mg/kg/day.

There was no effect of treatment an sperm motility, morphology or concentration. There were no treatment-related macroscopic findings.

Treatment related microscopic changes were seen in the nose/turbinates of all treated groups. Findings comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx, degeneration/regeneration/inflammatory cell infiltration of the epithelium and deformity/fusion of turbinates. In addition, epithelial hyperplasia of the non-glandular stomach was seen in occasional animals of both sexes given 500 and 1000 mg/kg/day and a single male given 250 mg/kg/day. A single case of erosion was seen in the non-glandular region of the stomach of a female given 1000 mg/kg/day, although these changes were of an uncertain relationship to treatment with TCD Alcohol DM.

Conclusion:

It was concluded, that for the assessment of systemic toxicity, oral administration of TCD Alcohol DM, an industrial chemical intermediate, to RccHanTM;WIST (Han Wistar) rats at dose levels of 250, 500 and 1000 mg/kg/day for 13 weeks the no-observed-adverse-effect-level (NOAEL) in this study was considered to be 1000 mg/kg/day.

It is acknowledged that adverse treatment related changes with TCD Alcohol DM were seen in the nose/turbinates of all treated groups of both sexes. Nasal findings comprised of inflammatory exudate in the lumen of the nasal cavity and nasopharynx, degeneration/regeneration/inflammatory cell infiltration of the epithelium and deformity/fusion of turbinates. This pathology was considered to be secondary to retrograde aspiration of material from the oropharynx into the nasopharynx and nasal cavity, this is considered to be a local effect, associated with the route of administration of this formulation and as such has been excluded from the assessment of systemic toxicity.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed

Dose descriptor:

NOAEL

1 000 mg/kg bw/day

Study duration:

subchronic

Species:

rat

Quality of whole database:

sufficient for assessment

Repeated dose toxicity: inhalation - systemic effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: inhalation

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

year of publication: 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented publication which meets basic scientific principles

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD TG 414

Principles of method if other than guideline:

Exposure to max. attainable vapor concentration of 1-octanol, 1-nonanol, and 1-decanol

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

CAS number IUPAC name
111-87-5 1-octanol
143-08-8 1-nonanol
112-30-1 1-decanol

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 0.5 m³ Hinners-type chamber
- Method of holding animals in test chamber: animals were placed in stainless steel wire mesh cages
- Source and rate of air: compressed air
- Method of conditioning air: vapor generation equipment, housed in a sealed glove box
- System of generating particulates/aerosols: a constant flow of alcohol was mixed with a known volume of heated compressed air. Th eresulting vapor/air mixture was introduced into the chamber airflow system upstream from the orifice plate. The resulting turbulence downstream from the orifice plate produced a uniform mixing of the test chemcial throughout the exposure chamber.
- Temperature in air chamber: 70-80°F (21-27°C)
- Air flow rate: no data
- Air change rate: no data
- Method of particle size determination: no data
- Treatment of exhaust air: no data

TEST ATMOSPHERE
- Brief description of analytical method used:
(1) infrared analyzer (Miran 1A) continously recorded concentrations near the rats breathing zone
() charcoal tube samples were collected 2 days per week and analyzed using gas chromatography. Spiked samples were also used to evaluate the accuracy of the GC results.
- Samples taken from breathing zone: yes

VEHICLE : no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

(1) infrared analyzer (Miran 1A) continously recorded concentrations near the rats breathing zone
(2) charcoal tube samples were collected 2 days per week and analyzed using gas chromatography. Spiked samples were also used to evaluate the accuracy of the GC results.

Duration of treatment / exposure:

gestation days 1-19
6 h/day (1-decanol)
7 h/day (1-octanol and 1-nonanol)

Frequency of treatment:

7/week

Remarks:

Doses / Concentrations:
400 (octanol), 150 (nonanol), 100 (decanol) mg/m³
Basis:
analytical conc.

No. of animals per sex per dose:

15

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: maximum concentrations that could be generated as a vapor at an average chamber temperature of 70-80°F. Generation of higher exposure concentrations resulted in aerosol production inside the inhalation chamber.

- Rationale for animal assignment: random

Positive control:

Yes; in total 13 alcohols with carbon chain skeltons of C1- to C10 were tested.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: group means reported

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: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: No data

CLINICAL CHEMISTRY: No data

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: no

Sacrifice and pathology:

For developmental toxicology evaluations dams were sacrificed on day 20 of gestation. Fetuses were removed, weighted, sexed and examined for external malformations. The frequency of visceral malformations and variations was determined in one half of the fetuses and the frequency of visceral malformations and variations in the other half.

Statistics:

One-way multivariate analysis of variance/ analysis of variance (MANOVA/ANOVA): number of corpora lutea, resorptions, male and female pups, mean male and female pup weight.
ANOVA: maternal weights, feed and water intake data,
ANOVA: fetal incidence data; also Variance Test for homogeneity of the binominal distribution; Kuskal-Wallis test was used if nonparametric analysis was more appropriate.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Critical effects observed:

not specified

Conclusions:

Maximum attainable vapor concentrations of aliphatic alcohols decrease rapidly withincreasing chain length. Max. inhalation concentration of 1-nonanol was 150 mg/m³. No maternal toxicity nor toxicity to reproduction was noted in dams exposed during gestation days 1-19 (7h/day). No developmental toxicity or teratogenicy was seen in the progeny.
In total 13 alcohols were tested, with carbon chain lengths ranging from C1 to C10. Maternal toxicity was only seen with C1 to C4 alcohols at ≥5000 ppm, but not with higher alcohols, probably due to the rapid decline of the vapor pressure with increasing chanin length. This implicates for isononanol (and other long chain alcohols):
(1) acute inhalation toxicity testing may scientifically be unjustified, due to low attainable vapor concentrations
(2) repeated inhalation toxicity testing: as above
(3) isononanol: repeated inhalation toxicity testing scientifically unjustified

Executive summary:

The effects of repeated inhalation exposure to 1-octanol, 1-nonanol, and 1-decanol was tested in female rats (15 per group) at the maximum attainable vapor concentrations of 400, 150, and 100 mg/m³, respectively. The rats were exposed during days 1 -19 of gestation for 6h (decanol) or 7 h/day (octanol and nonanol). Dams were sacrificed on gestation day 20, and dams and pups were examined for signs of maternal toxicity, toxicity to reproduction, and developmental toxicity.

(1) maximum attainable vapour concentrations at inhalation chamber temperatures between 21 -27°C were 400, 150, and 100 mg/m³ for octanol, nonanol, and decanol, respectively. Concentrations are bases on analytical data.

(2) No treatment related effects were seen

(2.1) dams:

- maternal toxicity: no signs of toxicity, no mortality, maternal body weight, mean food and mean water consumption unchanged compared to sham-treated controls

- toxicity to reproduction: mean numbers of corpora lutea/litter, resorptions/litter, male or female pups/litter all unchanged compared to sham-treated controls

(2.2) pups

- developmental toxicity: male and female pup weights unchanged compared to controls

- teratogenicity: frequency of skeletal or visceral malformations unchanged

(Nelson et al., Tox Ind Health 6:373 -387, 1990; Nelson et al., J Am Coll Tox 9:93 -97, 1990 (T04167 and T04176)

Overall

The vapour pressure of long chain aliphatic alcohols (carbon chain length C5 and more) is insufficient to reach toxic atmosphere concentrations. This is also valid for octahydro-4,7-methano-1H-indenedimethanol (TCD-Alcohol DM), a dialcohol with a carbon structure of C12. The vapour pressure is determined to be < 1 hPa (extrapolated from experimental result, see Sect. 4.6)

 

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: inhalation - local effects

Link to relevant study records

Reference

Endpoint:

repeated dose toxicity: inhalation

Remarks:

combined repeated dose and reproduction / developmental screening

Type of information:

migrated information: read-across based on grouping of substances (category approach)

Adequacy of study:

supporting study

Study period:

year of publication: 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well-documented publication which meets basic scientific principles

Reason / purpose for cross-reference:

reference to same study

Qualifier:

equivalent or similar to guideline

Guideline:

other: OECD TG 414

Principles of method if other than guideline:

Exposure to max. attainable vapor concentration of 1-octanol, 1-nonanol, and 1-decanol

GLP compliance:

not specified

Limit test:

no

Specific details on test material used for the study:

CAS number IUPAC name
111-87-5 1-octanol
143-08-8 1-nonanol
112-30-1 1-decanol

Species:

rat

Strain:

Sprague-Dawley

Sex:

female

Route of administration:

inhalation: vapour

Type of inhalation exposure:

whole body

Vehicle:

other: unchanged (no vehicle)

Details on inhalation exposure:

GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: 0.5 m³ Hinners-type chamber
- Method of holding animals in test chamber: animals were placed in stainless steel wire mesh cages
- Source and rate of air: compressed air
- Method of conditioning air: vapor generation equipment, housed in a sealed glove box
- System of generating particulates/aerosols: a constant flow of alcohol was mixed with a known volume of heated compressed air. Th eresulting vapor/air mixture was introduced into the chamber airflow system upstream from the orifice plate. The resulting turbulence downstream from the orifice plate produced a uniform mixing of the test chemcial throughout the exposure chamber.
- Temperature in air chamber: 70-80°F (21-27°C)
- Air flow rate: no data
- Air change rate: no data
- Method of particle size determination: no data
- Treatment of exhaust air: no data

TEST ATMOSPHERE
- Brief description of analytical method used:
(1) infrared analyzer (Miran 1A) continously recorded concentrations near the rats breathing zone
() charcoal tube samples were collected 2 days per week and analyzed using gas chromatography. Spiked samples were also used to evaluate the accuracy of the GC results.
- Samples taken from breathing zone: yes

VEHICLE : no

Analytical verification of doses or concentrations:

yes

Details on analytical verification of doses or concentrations:

(1) infrared analyzer (Miran 1A) continously recorded concentrations near the rats breathing zone
(2) charcoal tube samples were collected 2 days per week and analyzed using gas chromatography. Spiked samples were also used to evaluate the accuracy of the GC results.

Duration of treatment / exposure:

gestation days 1-19
6 h/day (1-decanol)
7 h/day (1-octanol and 1-nonanol)

Frequency of treatment:

7/week

Remarks:

Doses / Concentrations:
400 (octanol), 150 (nonanol), 100 (decanol) mg/m³
Basis:
analytical conc.

No. of animals per sex per dose:

15

Control animals:

yes, sham-exposed

Details on study design:

- Dose selection rationale: maximum concentrations that could be generated as a vapor at an average chamber temperature of 70-80°F. Generation of higher exposure concentrations resulted in aerosol production inside the inhalation chamber.

- Rationale for animal assignment: random

Positive control:

Yes; in total 13 alcohols with carbon chain skeltons of C1- to C10 were tested.

Observations and examinations performed and frequency:

CAGE SIDE OBSERVATIONS: Yes
- Time schedule: daily

DETAILED CLINICAL OBSERVATIONS: No data

BODY WEIGHT: Yes
- Time schedule for examinations: weekly

FOOD CONSUMPTION:
- Food consumption for each animal determined and mean daily diet consumption calculated as g food/kg body weight/day: group means reported

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: Yes
- Time schedule for examinations: weekly

OPHTHALMOSCOPIC EXAMINATION: No data

HAEMATOLOGY: No data

CLINICAL CHEMISTRY: No data

URINALYSIS: No data

NEUROBEHAVIOURAL EXAMINATION: no

Sacrifice and pathology:

For developmental toxicology evaluations dams were sacrificed on day 20 of gestation. Fetuses were removed, weighted, sexed and examined for external malformations. The frequency of visceral malformations and variations was determined in one half of the fetuses and the frequency of visceral malformations and variations in the other half.

Statistics:

One-way multivariate analysis of variance/ analysis of variance (MANOVA/ANOVA): number of corpora lutea, resorptions, male and female pups, mean male and female pup weight.
ANOVA: maternal weights, feed and water intake data,
ANOVA: fetal incidence data; also Variance Test for homogeneity of the binominal distribution; Kuskal-Wallis test was used if nonparametric analysis was more appropriate.

Clinical signs:

no effects observed

Mortality:

no mortality observed

Body weight and weight changes:

no effects observed

Food consumption and compound intake (if feeding study):

no effects observed

Food efficiency:

not specified

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

no effects observed

Ophthalmological findings:

not specified

Haematological findings:

not specified

Clinical biochemistry findings:

not specified

Urinalysis findings:

not specified

Behaviour (functional findings):

no effects observed

Organ weight findings including organ / body weight ratios:

not specified

Gross pathological findings:

no effects observed

Histopathological findings: non-neoplastic:

not examined

Histopathological findings: neoplastic:

not examined

Critical effects observed:

not specified

Conclusions:

Maximum attainable vapor concentrations of aliphatic alcohols decrease rapidly withincreasing chain length. Max. inhalation concentration of 1-nonanol was 150 mg/m³. No maternal toxicity nor toxicity to reproduction was noted in dams exposed during gestation days 1-19 (7h/day). No developmental toxicity or teratogenicy was seen in the progeny.
In total 13 alcohols were tested, with carbon chain lengths ranging from C1 to C10. Maternal toxicity was only seen with C1 to C4 alcohols at ≥5000 ppm, but not with higher alcohols, probably due to the rapid decline of the vapor pressure with increasing chanin length. This implicates for isononanol (and other long chain alcohols):
(1) acute inhalation toxicity testing may scientifically be unjustified, due to low attainable vapor concentrations
(2) repeated inhalation toxicity testing: as above
(3) isononanol: repeated inhalation toxicity testing scientifically unjustified

Executive summary:

The effects of repeated inhalation exposure to 1-octanol, 1-nonanol, and 1-decanol was tested in female rats (15 per group) at the maximum attainable vapor concentrations of 400, 150, and 100 mg/m³, respectively. The rats were exposed during days 1 -19 of gestation for 6h (decanol) or 7 h/day (octanol and nonanol). Dams were sacrificed on gestation day 20, and dams and pups were examined for signs of maternal toxicity, toxicity to reproduction, and developmental toxicity.

(1) maximum attainable vapour concentrations at inhalation chamber temperatures between 21 -27°C were 400, 150, and 100 mg/m³ for octanol, nonanol, and decanol, respectively. Concentrations are bases on analytical data.

(2) No treatment related effects were seen

(2.1) dams:

- maternal toxicity: no signs of toxicity, no mortality, maternal body weight, mean food and mean water consumption unchanged compared to sham-treated controls

- toxicity to reproduction: mean numbers of corpora lutea/litter, resorptions/litter, male or female pups/litter all unchanged compared to sham-treated controls

(2.2) pups

- developmental toxicity: male and female pup weights unchanged compared to controls

- teratogenicity: frequency of skeletal or visceral malformations unchanged

(Nelson et al., Tox Ind Health 6:373 -387, 1990; Nelson et al., J Am Coll Tox 9:93 -97, 1990 (T04167 and T04176)

Overall

The vapour pressure of long chain aliphatic alcohols (carbon chain length C5 and more) is insufficient to reach toxic atmosphere concentrations. This is also valid for octahydro-4,7-methano-1H-indenedimethanol (TCD-Alcohol DM), a dialcohol with a carbon structure of C12. The vapour pressure is determined to be < 1 hPa (extrapolated from experimental result, see Sect. 4.6)

 

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - systemic effects

Endpoint conclusion

Endpoint conclusion:

no study available

Repeated dose toxicity: dermal - local effects

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Repeated dose toxicity: oral

1 -Screening study

The repeated dose toxicity of octahydro-4,7-methano-1H-indenedimethanol (TCD Alcohol DM) can be accessed on basis of an oral combined repeated dose toxicity study (28 d) with a reproduction/developmental toxicity screening test (van Otterdijk/NOTOX, 2010; RL1, key study) under GLP conditions (OECD TG 422).

 

Octahydro-4,7-methano-1H-indenedimethanol (TCD Alcohol DM) was administered by daily oral gavage to male and female Wistar Han rats at dose levels of 150, 350 and 600 mg/kg/day. Males were exposed for 2 weeks prior to mating, during mating, and up to termination (for 28 days). The females were exposed for 2 weeks prior to mating, during mating, during post-coitum, and at least 4 days of lactation (for 42-53 days).

Formulation analysis showed that the formulations were prepared accurately and homogenously, and were stable for at least 6 hours at room temperature.

Parental results:

No toxicologically significant changes were noted in any of the parental parameters investigated in this study (i.e. clinical appearance, functional observations, body weights, food consumption, haematology and clinical biochemistry parameters, macroscopic examination, organ weights, and microscopic examination).

Reproductive/Developmental results:

No reproductive/developmental toxicity was observed at any dose level.

Based on these results, a parental, reproductive and developmental No Observed Adverse Effect Level (NOAEL) of 600 mg/kg/day was derived (van Otterdijk/NOTOX, 2010).

2 -Subchronic 90 -day study

Oral administration of TCD Alcohol DM, an industrial chemical intermediate, to RccHan™;WIST (Han Wistar) rats at dose levels of 250, 500 and 1000 mg/kg/day for 13 weeks was generally well tolerated without any systemic signs of toxicity.

Three females given 1000 mg/kg/day were killed for welfare reasons on Days 12, 13 and 80, due, in part, to breathing impairment; rats are obligate nasal breathers, the terminal clinical signs and changes observed at necropsy were consistent with ingestion of air, and these animals were recorded to have partially blocked nasal turbinates with reflux of dose having the potential to block nasal turbinates and may have contributed to the effects observed. The histopathological appearance and distribution of the findings in nose/nasal turbinates is suggestive of retrograde aspiration of material from the oropharynx via expiratory airflow into the nasopharynx and nasal cavity. This reflux after gavage dosing may be mechanically induced or occur spontaneously. Within the remaining animals on study, in-life signs of râles and rapid breathing were observed sporadically. Histopathology examination of surviving animals also demonstrated similar changes in the nose/nasal turbinates at all dose levels.

1. Mechanically induced reflux. This occurs directly after the gavage when test material adhering to the gavage tube is deposited in the oropharynx upon withdrawing the tube from the oesophagus. Additionally during tube removal gastric contents may also flow back into the oesophagus and into the oropharynx.

2. Spontaneous reflux. This can occur directly after removing the gavage tube or later when the animal is back in its cage. It may be related to gavage administration of a large volume, with resulting gastric overflow and may be exaggerated by pharmacological effects of the test item on gastro-oesophageal function.

The irritancy and viscosity of the test formulation have been shown to be critical in the occurrence of adverse respiratory effects after reflux from gavage dosing and the likelihood of gastric overflow is thought to be higher following administration of larger volumes of test material. (Damsch et Al)

The findings in the study of inflammatory exudate in the lumen of the nasal cavity and nasopharynx and degeneration/regeneration/inflammatory cell infiltration of the epithelium (which may have manifested as râles and rapid breathing in-life) are acute changes and considered to represent pathology due to recent nasal reflux. Deformity/fusion of turbinates is a chronic finding and is considered to be a sequelae to the acute inflammatory changes and its presence suggests an episode of the initiating reflux had occurred in previous days/weeks. The presence in some individuals of only the turbinate deformity/fusion without the acute

inflammatory changes suggests the reflux was not recent and animals with a combination of turbinate deformity/fusion together with acute inflammatory changes suggests that the reflux happened on more than one occasion in that individual.

The findings in the stomach could be consistent with mild irritation. Epithelial hyperplasia and erosion of the non-glandular stomach was seen in individuals of most treated groups.The low numbers affected and lack of consistency between the two sexes made this finding have an uncertain relationship to treatment. The cause of the haematological (reduction in activated partial thromboplastin time in males and females receiving 500 or 1000 mg/kg/day) and biochemical (slightly high alkaline phosphatase activities in males receiving 1000 mg/kg/day, slightly high cholesterol concentration in males receiving 1000 mg/kg/day) changes in the blood was not established.

These changes are typical of an effect on the liver or kidney. Although high liver weights were observed in males and females at 500 or 1000 mg/kg/day and high kidney weight in males and females given 1000 mg/kg/day, there were no significant histopathological

changes in the liver or kidney. There was no histopathological correlate for the high spleen weights in males given 1000 mg/kg/day. Consequently these findings were not considered to be toxicologically important. It should be noted that there were no adverse effects noted on the male and female reproductive organs, oestrous cycle, and sperm parameters at any dose up to and including 1000 mg/kg bw and day.

 

Repeated dose toxicity: inhalation

Nelson et. al. (1990; supporting study) have investigated series of alcohols showing that alcohols with a chain length from C5 on upward do not exert toxic effects due to low atmosphere concentrations achievable (Journal of the American College of Toxicology 1990 (9): 93-97; Toxicol and Industrial Health 1990 (6): 373-387).

The carbon frame of TCD alcohol DM consists of 12 carbon atoms. In addition, the second hydroxyl group will further lower the volatility of TCD Alcohol DM. The vapour pressure is calculated to be < 1 hPa (Sect. 4.6). Atmosphere concentrations attainable under ambient conditions are assessed to be too low to exert toxic effects.

A subchronic inhalation study was, therefore, not deemd necessary, in accordance with Annex IX No. 8.6.2, column 2.

Justification for selection of repeated dose toxicity via oral route - systemic effects endpoint:
GLP guideline study with high reliability (Klimisch score 1)

Justification for selection of repeated dose toxicity inhalation - systemic effects endpoint:
Supporting study provides evidence that the vapour pressure is too low to reach air concentrations of toxicological relevance (read-across to supporting subtance (structural analogues - octanol, nonanol, decanol).

Justification for selection of repeated dose toxicity inhalation - local effects endpoint:
Supporting study provides evidence that the vapour pressure is too low to reach air concentrations of toxicological relevance.

Justification for selection of repeated dose toxicity dermal - systemic effects endpoint:
A reliable repeated dose toxicity study for the relevant route of exposure is available, thus no dermal repeated toxicity study is required.

Justification for selection of repeated dose toxicity dermal - local effects endpoint:
A reliable repeated dose toxicity study for the relevant route of exposure is available, thus no dermal repeated toxicity study is required.

Justification for classification or non-classification

Criteria set in Regulation (EC) No 1272/2008 for specific target organ toxicity - repeated exposure are not met. Classification is not required.