Registration Dossier

Administrative data

Description of key information

- Oral LD50, rats - LD50 between 300 and 2000 mg/kg.  LD50 reported as 700 mg/kg.

- Dermal LD50, rabbits - LD50 = 1700 mg/kg.

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
4 (not assignable)
Rationale for reliability incl. deficiencies:
other: No claim of GLP compliance was made, no reference to official test guidelines was contained the in the report and the details of the testing laboratory were not supplied. On this basis, reliability of the data could not be properly asessed..
Reason / purpose:
reference to other study
Qualifier:
no guideline followed
Principles of method if other than guideline:
Test groups of ten rats per dose level were administered a single oral dose of the test material. Seven male dose groups (six treated groups in the range 0.541 - 1.287 mL/kg plus one control group) and eight female dose groups (seven treated groups in the range 0.568 - 1.566 mL/kg plus one control group) were tested. The rats were fasted the night before dosing, and for three hours afterwards. The rats were observed for 14 days after dosing, and a post-mortem examination was conducted following sacrifice of the surviving rats after the observation period.
GLP compliance:
no
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: four weeks old
- Fasting period before study: one night prior to dosing
- Housing: cage
- Diet (e.g. ad libitum): ad libitum
- Water (e.g. ad libitum):ad libitum
- Acclimation period: one week


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 24 +/- 2
- Humidity (%): 45-85



Route of administration:
oral: gavage
Details on oral exposure:
MAXIMUM DOSE VOLUME APPLIED: 1.566 ml/kg (female); 1.287 ml/kg (male)


DOSAGE PREPARATION (if unusual): Dose was adjusted by volume only. Each animals received a different volume of undiluted test material
Doses:
Male: 0.541, 0.660, 0.755, 0.849, 1.089, 1.287 ml/kg. Female: 0.568, 0.638, 0.795, 0.899, 1.001, 1.325, 1.566 ml/kg.

Nb - the above doses, expressed in terms of mass of test material/kg bodyweight are:
Males; 511, 623, 713, 801, 1028, and 1215 mg/kg bw
Females; 536, 602, 750, 849, 945, 1251, and 1478 mg/kg bw.
No. of animals per sex per dose:
10
Control animals:
yes
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: daily
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
male
Dose descriptor:
LD50
Effect level:
700 mg/kg bw
Sex:
female
Dose descriptor:
LD50
Effect level:
780 mg/kg bw
Clinical signs:
Decline of spontaneous movement was noted by some hours after the administration and subsequently blepharoptosis, diarrhea, and reddish brown vomit or flow of tears were observed. These syptoms disappeared 6-8 days later.
Other findings:
As to rats that died during the experiment, intense ulceration or necrosis was observed on the stomach and intestinal canal. Particularly, such sympton the stomach was remarkable and the higher dose resulted in the severer symptom. As to other rats that lived 14 days, the accretion of stomach with liver was observed in the female group of 0.795 ml/kg dose and all male groups of lower doses to higher doses. Definite Dose-Response relationship, however, was not found. No extreme change was observed on any other organs.
Conclusions:
The acute oral LD50 rats: 700 mg/kg (male) and 780 mg/kg (female)
Executive summary:

LD50 values of 1,3-BAC to rats under the condition that undiluted 1,3-BAC is orally administered to the rats of 105 ± 3.7g and 88 ± 3.9g in body weight ± standard deviation. Male 0.74 ml/kg (0.65-0.82 ml/kg, P=0.05) or 700 mg/kg (610-770 mg/kg, P=0.05) Female: 0.83 ml/kg (0.71-0.94 ml/kg, P=0.05) or 780 mg/kg (670-890 mg/kg, P=0.05).

Endpoint:
acute toxicity: oral
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
29 July 1997 to 20 August 1997
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: Study performed according to GLP and internationally accepted test guidelines.
Qualifier:
according to
Guideline:
OECD Guideline 401 (Acute Oral Toxicity)
Qualifier:
according to
Guideline:
EU Method B.1 (Acute Toxicity (Oral))
Principles of method if other than guideline:
On occasions the relative humidity was above the limit specified in the study protocol (70%). This deviation was considered not to affect the purpose or integrity of the study.
GLP compliance:
yes (incl. certificate)
Test type:
fixed dose procedure
Limit test:
yes
Species:
rat
Strain:
Sprague-Dawley
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: eight to twelve weeks of age
- Weight at study initiation: male: 202 to 241g and females:202 to 225g
- Fasting period before study: overnight and 3 hours after dosing
- Housing: The animals were housed in groups of up to five by sex in solid-floor polypropylene cages furnished with woodflakes
- Diet: ad libitum
- Water: ad libitum
- Acclimation period:5 days


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19°C to 24
- Humidity (%): 58 to 73
- Air changes (per hr): 15 changes per hour
- Photoperiod (hrs dark / hrs light): 12/12
Route of administration:
oral: gavage
Vehicle:
water
Details on oral exposure:
VEHICLE
- Concentration in vehicle: 20 mg/mL
- Amount of vehicle (if gavage): 10 mL/kg

MAXIMUM DOSE VOLUME APPLIED: 200 mg/kg

Doses:
All animals were dosed once only by gavage using a metal cannula attached to a graduated syringe. The volume administered to each animal was calculated according to its fasted bodyweight at the time of dosing.
No. of animals per sex per dose:
5 Male and 5 Female rats treated per dose
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: 30 min, 1 hr, 2hrs, 4 hrs then once daily for 14 days
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight.
Preliminary study:
Range-finding study: The two animals treated with the undiluted test material at a dose of 2000 mg/kg were found dead four hours or one day after dosing. Clinical signs of toxicity noted were ataxia, pallor of the extremities, hunched posture, lethargy, pilo-erection, ptosis, decreased respiratory rate and laboured respiration.
When the test material was administered as a formulation, animals treated with 2000 mg/kg were found dead one day after dosing. Clinical signs of toxicity noted in theses animals were ataxia, pallor of the extremities, hunched posture, lethargy, pilo-erection, ptosis, decreased respiratory rate, laboured and noisy respiration. There were no deaths or clinical signs of toxicity noted in animals treated with 200 mg/kg.
The formulated doses were prepared at 200 or 20 mg/mL (as appropriate) in distilled water, and dosed at a fixed volume of 10 mL/kg.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 200 - < 2 000 mg/kg bw
Remarks on result:
other: No Mortality at 200 mg/kg; 100% mortality at 2000 mg/kg during range-finding test.
Mortality:
In the preliminary (range-finding) study, all animals dosed at 2000 mg/kg died. No animals dosed at 200 mg/kg died, and so this level (200 mg/kg) was used in the main study.
No mortality was observed in main study in animals observed 14 days after dose.
Clinical signs:
One female, from day 9 onward was observed to have noisy respiration. One female, one day after dosing was observed to have hunched posture and noisy respiration.
Body weight:
All animals showed an expected gain in bodyweight during the study.
Gross pathology:
No abnormalities detected
Interpretation of results:
harmful
Remarks:
Migrated information Criteria used for interpretation of results: EU
Conclusions:
The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley CD strain rat was found to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight.
Executive summary:

A study was performed to assess the acute oral toxicity of the test material in the Sprague-Dawley CD strain rat. The method followed that in the OECD Guidelines for Testing of Chemicals No. 401 "Acute Oral Toxicity" (adopted 24 February 1987) and Method B1 of Commission Directive 92/69/EEC (which constitues Annex V of Council Directive 67/548/EEC).

Following a range-finding study, a group of ten fasted animals (five males and five females) was given a single oral dose of test material as a solution in distilled water at a dose level of 200 mg/kg bodyweight. The animals were observed for fourteen days after the day of dosing and were then killed and subjected to gross pathological examination.

There were no deaths. Clinical signs of toxicity noted in two females were hunched posture and/or noisy respiration. No other signs of toxicity were noted.

All animals showed expected gain in bodyweight during the study.

No abnormalities were noted at necropsy.

The acute oral median lethal dose (LD50) of the test material in the Sprague-Dawley CD strain rat was found to be greater than 200 mg/kg bodyweight but less than 2000 mg/kg bodyweight. This supports the findings of the key study (Kashima Labs, 2007).

Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
700 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
no study available

Acute toxicity: via dermal route

Link to relevant study records

Referenceopen allclose all

Endpoint:
acute toxicity: dermal
Type of information:
experimental study
Adequacy of study:
supporting study
Reliability:
3 (not reliable)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
No claim of GLP complaince or adherence with an official test guideline is made in the report. Comparison of the test methodology used to modern OECD guidelines highlights a number of significant discrepancies between the method employed in the test and the modern guideline, and the study report was considered generally not sufficiently detailed to consider the study data reliable. Note that as the material is classified as corrosive to skin (refer to dossier section 7.3), further acute toxicity testing cannot be justified, and so the result of this study is the only one available for acute toxicity testing by the dermal route. On this basis, this study was considered the key study, in spite of the poor reliability of the study.
Qualifier:
no guideline followed
Principles of method if other than guideline:
Animals: Sixteen albino rabbits of the New Zealand variety were obtained from the Skippack Farms and individually housed in suspended cages. All rabbits were fed Purina Rabbit Pellets and tap water on an ad libitum basis
Preparation and Treatment: Following a brief acclimation period, and prior to treatment, the backs of all rabbits were clipped free of fur; the animals were assigned to four treatment groups. Two rabbits in each group were abraded with a burred needle and two remained intact. Treatment consisted of a single dermal application of the 1,3-bis aminomethyl cyclohexane (1,3-BAC) to the skin. The test site was completely occluded with an impervious dam. After 24 hours, the wrapping was removed and the residue gently washed off.
Laboratory Observations
Animals were examined daily for abnormal behaviour or mortality. Skin reactions of erythema and edema were scored at 24 hours and daily thereafter for 14 days according to the method of Draize. Body weights were recorded on Days 0, 7, and 14. At termination or death, each animal was subjected to a gross examination of the viscera, except in cases of advanced autolysis of the tissue.

Note that according to OECD test guideline 402, skin should not be abraded and only animals with intact skin should be used. This guideline also recommends the use of at least 5 animals per dose level, unless a reason for the use of fewer animals can be justified. As half of the animals in each dose level had abraded skin, the results obtained from these animals should not be considered, as abrasion of the skin is likely to affect absorbtion of the test material, and as only two animals at each level had unabraded skin the sample size for each dose level is substantially smaller than the guideline recommendation. As a result of these aspects, this study must be consodered less reliable than one conducted according to the guideline.
GLP compliance:
not specified
Test type:
standard acute method
Species:
rabbit
Strain:
New Zealand White
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Age at study initiation: not specified
- Weight at study initiation: 2.77-3.90 kg
- Fasting period before study: not specified
- Housing: suspended cages
- Diet: ad libitum- Purina Rabbit Pellets
- Water: ad libitum

Type of coverage:
occlusive
Vehicle:
unchanged (no vehicle)
Details on dermal exposure:
TEST SITE
- Area of exposure: not specified
- Type of wrap if used: "impervious dam"

REMOVAL OF TEST SUBSTANCE
- Washing (if done): yes, with water
- Time after start of exposure: 24 hours

TEST MATERIAL
- Concentration (if solution): 1260, 1580, 2000, 2510 mg/kg
- Constant volume or concentration used: yes


Duration of exposure:
24 hours
Doses:
1260, 1580, 2000, 2510 mg/kg
No. of animals per sex per dose:
4 animals per dose, 2 with abraded skin
Control animals:
not specified
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing:Daily
- Necropsy of survivors performed: yes
- Other examinations performed: clinical signs, body weight
Sex:
not specified
Dose descriptor:
LD50
Effect level:
1 700 mg/kg bw
95% CL:
1 417 - 2 040
Mortality:
1260 mg/kg: no mortality; 1580 mg/kg: one death day 2, one death day 3; 2000 mg/kg:one death day 2, one death day 3; 2510 mg/kg: one death day 1, two death day 2, one death day 4.
Body weight:
Bodyweight tended to decrease in animals exposed above 1580 mg/kg.
Interpretation of results:
harmful
Remarks:
Migrated information According to Directive 67/548/EEC. Criteria used for interpretation of results: EU
Conclusions:
The dermal LD50 was calculated to be 1700 mg/kg of body weight.* The 95% confidence limits are 1417 - 2040 mg/kg. 
Executive summary:

An acute dermal toxicity study was conducted by Huntingdon Research Centre, registered in Maryland, USA, on behalf of Sherwin-Williams Chemicals, Ohio, USA, to determine the acute toxic effects by dermal exposure of the test substance 1,3 -Bis Aminomethyl Cyclohexane.

Four groups of two rabbits with unabraded skin and two with abraded skin were exposed to the test material for a period of 24 hours, then the dressing removed and the exposure site washed. Dose levels assessed were 1260, 1580, 2000, and 2510 mg/kg. The exposure sites were observed for signs of skin reactions at 24 hours, and daily thereafter for 14 days. Animals were observed for clinical signs and mortality for 14 days.

After 24 hours of exposure, all rabbits (intact and abraded) exhibited severe erythema and edema. Within a few days, the edema subsided and the skin became "leathery" in appearance and to the touch. All surviving rabbits tended to lose weight during the 14-day post-exposure period. All rabbits died at the 2510 mg/kg level, 2/4 rabbits died at each of the 2000 and 1580 mg/kg levels, and all rabbits at the 1260 mg/kg level survived until the end of the 14 -day observation period.

The dermal LD50 was calculated to be 1700 mg/kg of body weight.

Endpoint:
acute toxicity: dermal
Data waiving:
study scientifically not necessary / other information available
Justification for data waiving:
other:
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
1 700 mg/kg bw
Quality of whole database:
It is noted that the study on which the endpoint conclusion is based was assigned a reliability (Klimisch) score 3, which would not typically be considered sufficient to satisfy data requirements under REACH. In the case of 1,3-BAC it must be considered that the substance is corrosive to skin (note that the substance is classified as Skin Corrosive Category 1A according to the CLP Regulation); therefore new acute dermal toxicity testing cannot be conducted on ethical / animal welfare grounds.

On the basis of a number of inconsistencies between the methodology used in the only available study and current official guidelines, the overall database for Acute Dermal Toxicity cannot be considered reliable.

Additional information

Acute oral toxicity:

Three studies are available regarding the assessment of acute oral toxicity of the test substance, 1,3-Bis(aminomethyl)cyclohexane; one study performed in 2007, one in 1998, and a further study in 1980. The studies performed in 2007 and 1998 are considered reliable, both having been performed according to recognised test guidelines, and in compliance with GLP. The reliability of the third study (1980) could not be assessed as only a limited report was available, and the level of detail in the report is insufficient to determine reliability. In each of the 2007 and 1998 studies, 100% mortality was seen at 2000 mg/kg, and no mortality was seen at 300 mg/kg and 200 mg/kg, respectively. Each of these results is adequate for classification purposes, however as neither study report concluded an LD50value, the value reported in the third (1980) study for the acute oral LD50(700 mg/kg for males) in rats will be used.

It was noted that when the effect of the test material administered as neat substance was compared to the effect when administered as a formulation (i.e. a diluted form), the neat material caused more severe toxic effects and resulted in a lower LD50than when administered in a diluted formulation. As this effect was only noted in the preliminary test of the 1998 study, and only tentatively explored in the 1980 study (test material was administered to mice as a formulation as it could not be accurately dosed as pure substance, however one group was dosed with pure substance for comparison), the effect was not accurately quantified.

Acute dermal toxicity:

An acute dermal toxicity study was conducted to determine the acute toxic effects by dermal exposure of the test substance 1,3 -Bis Aminomethyl Cyclohexane.

Four groups of two rabbits with unabraded skin and two with abraded skin were exposed to the test material for a period of 24 hours, then the dressing removed and the exposure site washed. Dose levels assessed were between 1260 and 2510 mg/kg. The exposure sites were observed for signs of skin reactions at 24 hours, and daily thereafter for 14 days. Animals were observed for clinical signs and mortality for 14 days.

After 24 hours of exposure, all rabbits (intact and abraded) exhibited severe erythema and edema. Within a few days, the edema subsided and the skin became "leathery" in appearance and to the touch. All surviving rabbits tended to lose weight during the 14-day post-exposure period. All rabbits died at the 2510 mg/kg level, 2/4 rabbits died at each of the 2000 and 1580 mg/kg levels, and all rabbits at the 1260 mg/kg level survived until the end of the 14 -day observation period.

The dermal LD50was calculated to be 1700 mg/kg of body weight.

It was noted that the acute dermal toxicity study described above would not generally be considered sufficiently reliable for use as a key study (it was judged to be a Klimisch score 3 for reliability), however as the material is classified is corrosive (note results in section 7.3), testing for acute dermal or acute inhalation toxicity would not generally need to be conducted (as stated in Annex VIII of the REACH regulation). On this basis, additional testing for acute toxicity by inhalation or by dermal route would not be recommended, and the result from the Huntingdon Research Centre study will be used for classification purposes, and for the calculation of the relevant DNELs.


Justification for selection of acute toxicity – oral endpoint
Although the nominated study (1980) was not as reliable as the other available studies (2007 and 1998), the 2007 and 1998 studies did not conclude an exact value for the LD50 (rather, a range was indicated for each study). The value for the LD50 concluded in the 1980 study is consistent with the conclusions of the other two studies; on this basis the LD50 value in rats is used as the dose descriptor for the CSA.

Justification for classification or non-classification

Each of three acute oral toxicity studies in rats concluded that the LD50 value was in the range 200 - 2000 mg/kg, and two of these studies confirmed that the LD50 value was in the range 300 - 2000 mg/kg. According to the GHS system (EC Regulation 1272/2008 used for the purposes of this classification), the substance should be classified as acute oral toxicity category 4.

The acute dermal toxicity was assessed and the LD50 value was found to be 1700 mg/kg. On this basis, the test substance should be classified as acute dermal toxicity category 4 according to GHS (as described in EC regulation 1272/2008).

With respect to classification for Specfic Target Organ Toxicity following a Single Exposure (STOT SE), treatment-related effects were observed at necropsy in the stomachs of rats treated orally with the substance at 2000 mg/kg (Kashima Labs, 2007). It was noted that the level at which these effects were observed was lethal to all tested animals; no such effects were seen at the next lowest dose level (300 mg/kg). As it cannot be confirmed that these effects occur at sub-lethal doses it is considered that the substance does not meet the criteria for classification as STOT SE. It is considered likely that the effects seen in the stomach of the deceased animals is related to the effect causing the mortality, for which the substance has been classified as acute category 4, as noted above.

It was noted that sub-lethal effects (oedema and erythema, followed by "leathery" appearance of the skin following the subsidance of the oedema and erythema) were seen in all animals dosed in the Acute Dermal Toxicity study. It is considered that these effects are due to the corrosive nature of the substance; because 1,3-BAC is already classified for Skin Corrosion (please refer to dossier section 7.3) a supplementary classification for specific target organ toxicity following a single dermal exposure (STOT SE, dermal route) is not justified.