Registration Dossier

Administrative data

Description of key information

Oral
Rat; LD50 ca. 1276.1 mg/kg (standardized test protocol) [Smyth et al. 1969]
Dermal
Rabbit: LD50 ca. 452.5 mg/kg (standardized test protocol) [Smyth et al. 1969]; LD50 > 3420 mg/kg bw (MDACH salt) [BASF, 2009]
Inhalation
Rat: 8 hours IHT (standardized test protocol): No death occurred when the test animals were exposed to an atmosphere saturated with vapours of test substance [Smyth et al. 1969]

Key value for chemical safety assessment

Acute toxicity: via oral route

Link to relevant study records
Reference
Endpoint:
acute toxicity: oral
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study meets generally accepted scientific principles (restriction: purity unknown, limited documentation). The registered substance is a reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen (common name: diaminomethylcyclohexane or MDACH, respectively) consisting of the two isomers 4 -methylcyclohexane-1,3-diamine (CAS 13897-55-7; molecular weight approx. 128 g/mol) and 2-methylcyclohexane-1,3-diamine (CAS 13897-56-8; molecular weight approx. 128 g/mol). The registered substance was tested in several toxicity studies, which were conducted on the one hand using 4-methylcyclohexane-1,3-diamine (CAS 13897-55-7) which is the main isomer of the registered substance (up to 90%) and on the other hand the reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen was tested as such (for details see read across justification, IUCLID chapter 13).
Qualifier:
no guideline followed
Principles of method if other than guideline:
Estimation of single oral dose toxicity of the test substance by the gastric intubation of groups of male rats.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: Carworth-Wistar
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: own breeding
- Age at study initiation: 4-6 weeks
- Weight at study initiation: 90-120 g
- Fasting period before study: no
- Diet (e.g. ad libitum): Rockland rat diet


ENVIRONMENTAL CONDITIONS
not reported
Route of administration:
oral: gavage
Vehicle:
water
Remarks:
not specified: water or corn oil
Details on oral exposure:
VEHICLE
No further information provided.
Doses:
No data
No. of animals per sex per dose:
5
Control animals:
not specified
Details on study design:
Animals were observed for signs of toxicity after dosing, and throughout the 14-day observation interval. Animals were weighed prior to dosing and at the end of the observation interval. Animals that died on study were subjected to necropsy; surviving animals were sacrificed at the end of the observation interval and necropsied.
Statistics:
Thompson's method of calculating the LD50 was applied to the 14-day mortality data.
Sex:
male
Dose descriptor:
LD50
Effect level:
ca. 1 276.1 mg/kg bw
Based on:
test mat.
Remarks on result:
other: corresponding to a LD50 of 1.41 mL, calculated with a density of 0.905 g/mL as prefered result of the OECD SIDS
Mortality:
Not reported in the publication.
Clinical signs:
Not reported in the publication.
Body weight:
Not reported in the publication.
Gross pathology:
Not reported in the publication.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed
Dose descriptor:
LD50
Value:
1 276.1 mg/kg bw

Acute toxicity: via inhalation route

Link to relevant study records
Reference
Endpoint:
acute toxicity: inhalation
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Study meets generally accepted scientific principles (IHT; restriction: purity unknown, limited documentation, no determination of test atmosphere concentration). The registered substance is a reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen (common name: diaminomethylcyclohexane or MDACH, respectively) consisting of the two isomers 4 -methylcyclohexane-1,3-diamine (CAS 13897-55-7; molecular weight approx. 128 g/mol) and 2-methylcyclohexane-1,3-diamine (CAS 13897-56-8; molecular weight approx. 128 g/mol). The registered substance was tested in several toxicity studies, which were conducted on the one hand using 4-methylcyclohexane-1,3-diamine (CAS 13897-55-7) which is the main isomer of the registered substance (up to 90%) and on the other hand the reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen was tested as such (for details see read across justification, IUCLID chapter 13).
Qualifier:
no guideline followed
Principles of method if other than guideline:
Inhalation hazard test: to evaluate the acute inhalation toxicity, group of male Carworth-Wistar rats were exposed for 8 hours to an atmosphere saturated with vapours of the test substance and observed 14 days for mortality.
GLP compliance:
no
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
other: Sherman
Sex:
not specified
Details on test animals and environmental conditions:
TEST ANIMALS
- Weight at study initiation: usually 100-150 g


ENVIRONMENTAL CONDITIONS
not reported
Route of administration:
inhalation: vapour
Type of inhalation exposure:
whole body
Vehicle:
air
Details on inhalation exposure:
GENERATION OF TEST ATMOSPHERE / CHAMBER DESCRIPTION
- Exposure apparatus: Sealed chamber
- Exposure chamber volume: 120-liter
- Method of holding animals in test chamber: Six rats are rapidly introduced by means of a drawer-type cage designed to minimize vapor loss.
- Source and rate of air: For inhalation periods of ten, five and two minutes in duration, a static technique is used whereby 50 to 100 grams of material, spread over a shallow tray 200 square inches in area, is placed in a 120-liter sealed chamber for at least 24 hours.
- Method of conditioning air: The vapor-air mixture is generated by passing 2.5 liters/minute of dried air at room temperature through a fritted glass disc immersed to a depth of at least one inch in approximately 50 ml of the test chemical contained in a gas-washing bottle.
The exposure apparatus is described in sufficient detail in Smyth et al. 1949, "Assay for Acute Vapor Toxicity".
Analytical verification of test atmosphere concentrations:
no
Duration of exposure:
8 h
Concentrations:
Not determined (IHT)
No. of animals per sex per dose:
6
Control animals:
not specified
Details on study design:
Animals were weighed prior to exposure and at the end of the post-exposure 14-day observation interval. Animals were observed for signs of toxicity during and after the exposure interval, and daily during the 14-day postexposure observation interval.
Autopsies are performed on all of the rats to ascertain that they did not die of extraneous infection. Gross pathologic change is recorded and liver, spleen, adrenal gland, kidney, and testis are taken for histopathological interpretation from 2 of the rats which died during the 14 day observation period and also from 2 that survived. The survivors are weighed and killed by transecting the spinal cord in the cervical region. An attempt is made to take the tissues for histopathological study from rats exposed to the concentration which killed either 2, 3, or 4 of the 6 exposed.
Statistics:
The LC50 value was calculated ex post with a probit analysis according to Finney.
Sex:
male/female
Dose descriptor:
LC50
Based on:
test mat.
Exp. duration:
8 h
Remarks on result:
other: No death occurred when the test animals were exposed to an atmosphere saturated with vapours of test substance (IHT)
Mortality:
No death occurred
Clinical signs:
other: Not reported in the publication.
Body weight:
Not reported in the publication.
Gross pathology:
Not reported in the publication.
Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Quality of whole database:
only limited details on test conditions are available and the concentration of the test atmosphere was not determined; no deaths were observed up to the end of the observation period

Acute toxicity: via dermal route

Link to relevant study records
Reference
Endpoint:
acute toxicity: dermal
Type of information:
migrated information: read-across from supporting substance (structural analogue or surrogate)
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: see 'Remark'
Remarks:
Guideline study (GLP). The registered substance is a reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen (common name: diaminomethylcyclohexane or MDACH, respectively) consisting of the two isomers 4 -methylcyclohexane-1,3-diamine (CAS 13897-55-7; molecular weight approx. 128 g/mol) and 2-methylcyclohexane-1,3-diamine (CAS 13897-56-8; molecular weight approx. 128 g/mol). The registered substance was tested in several toxicity studies, which were conducted on the one hand using 4-methylcyclohexane-1,3-diamine (CAS 13897-55-7) which is the main isomer of the registered substance (up to 90%) and on the other hand the reaction product of 2,4-dinitrotoluene and 2,6-dinitrotoluene and hydrogen was tested as such (for details see read across justification, IUCLID chapter 13).
Qualifier:
according to
Guideline:
OECD Guideline 402 (Acute Dermal Toxicity)
Version / remarks:
February 24, 1987
Deviations:
no
Qualifier:
according to
Guideline:
EPA OPPTS 870.1200 (Acute Dermal Toxicity)
Version / remarks:
August 1998
Qualifier:
according to
Guideline:
other: Japan MAFF Testing Guideline of 12 Nosan No. 8147 as this in line with OECD 402.
GLP compliance:
yes
Test type:
standard acute method
Limit test:
no
Species:
rat
Strain:
Wistar
Sex:
male/female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: rat / Wistar / Crl:WI (Han) SPF from Charles River Wiga GmbH, Sandhofer Weg 7, 97633 Sulzfeld, Germany
- Age at study initiation: young adult animals (male animals approx. 8 weeks, female animals approx. 12 weeks)
- Weight at study initiation: animals of comparable weight (± 20% of the mean weight); 200.8-205.4 g in females; 226.6-241.8 g in males
- Housing: single housing in Makrolon cage, type III
- Diet (e.g. ad libitum): VRF1(P); SDS Special Diets Services, 67122 Altrip, Germany); ad libitum
- Water (e.g. ad libitum): tap water; ad libitum
- Acclimation period: at least 5 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22-26°C
- Humidity (%): 20-80%
- Photoperiod (hrs dark / hrs light): 12 h / 12 h (6.00 a.m. – 6.00 p.m. / 6.00 p.m. – 6.00 a.m.)
Type of coverage:
semiocclusive
Vehicle:
water
Remarks:
doubly distilled water
Details on dermal exposure:
TEST SITE
- Area of exposure: ca. 40 cm² of the clipped epidermis (dorsal and dorsolateral parts of the trunk)
- % coverage: corresponds to at least 10% of the body surface
- Type of wrap if used: covering of the application site with a semi occlusive dressing (the bandage consists of four layers absorbent gauze, Ph. Eur. Lohmann GmbH & Co. KG and Fixomull stretch (adhesive fleece), Beiersdorf AG)

REMOVAL OF TEST SUBSTANCE
- Washing (if done): rinsing of the application site with warm water
- Time after start of exposure: 24 hours after test substance application

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 10 ml/kg bw
- Concentration (if solution): 17.10 and 34.20 g/100 ml, respectively for the low and high dose groups
- Constant volume or concentration used: yes
Duration of exposure:
24 hours
Doses:
1710 and 3420 mg/kg bw
No. of animals per sex per dose:
5
Control animals:
not required
Details on study design:
- Duration of observation period following administration: 14 days
- Frequency of observations and weighing: recording of clinical signs several times on the day of administration, and at least once daily thereafter each workday for the individual animals. Determination of individual body weights shortly before administration (day 0), weekly thereafter and on the last day of observation. Check for any dead or moribund animal at least once each workday.
- Necropsy of survivors performed: yes; necropsy with gross-pathology examination on the last day of the observation period after sacrifice with CO2 in a chamber with increasing concentrations over time
- Other examinations performed: scoring of skin findings according to Draize by individual readings 30 – 60 minutes after removal of the semi-occlusive dressing (day 1), weekly thereafter, and on the last day of observation.
Statistics:
Calculations were performed using Microsoft Excel 2003 and checked with a calculator.
Sex:
male/female
Dose descriptor:
LD50
Effect level:
> 3 420 mg/kg bw
Based on:
test mat.
Remarks on result:
other: no mortality occurred and no clinico-pathological effects were observed up to this dose level
Mortality:
No mortality occurred.
Clinical signs:
No systemic clinical signs were observed during clinical examination.
Body weight:
The mean body weight of the animals increased within the normal range throughout the study period in both dosage groups.
Mean body weight of the female animals were constant during the first post-exposure observation week but increased during the second week within the normal range in
both dosage groups.
Gross pathology:
No macroscopic pathologic abnormalities were noted in the animals (10 males and 10 females) examined on the last day of observation.
Other findings:
- Local effects: no local effects were observed.

Table 1: Body weight changes

Days after treatment

Mean body weight changes ± standard deviations

1710 mg/kg bw

3420 mg/kg bw

Males

Females

Males

Females

0

241.8 ± 8.73

200.8 ± 6.26

226.6 ± 5.81

205.4 ± 4.34

7

256.6 ± 10.31

202.4 ± 7.02

247.4 ± 10.95

209.8 ± 1.79

15

277.6 ± 13.20

209.4 ± 7.83

273.6 ± 14.71

218.8 ± 3.11

 

Endpoint conclusion
Endpoint conclusion:
no adverse effect observed
Dose descriptor:
LD50
Value:
3 420 mg/kg bw
Quality of whole database:
acute dermal toxicity study conducted according to the OECD guideline 402; no mortality was oberserved

Additional information

Acute oral toxicity

Groups of five male Carworth-Wistar rats were treated with the test substance under standardized conditions at dose levels not specified, and were observed for 14 days (Smyth et al. 1969; Rel 2). The reported acute oral LD50 was approx. 1276.1 mg/kg bw. No further details were available.

 

Acute inhalation toxicity

To evaluate the acute inhalation toxicity, group of 6 male Carworth-Wistar rats were exposed for 8 hours to an atmosphere saturated with vapours of the test substance and observed 14 days for mortality (Smyth et al. 1969; Val 2). Whereas only limited details on test conditions are available and the concentration of the test atmosphere was not determined, no deaths were observed up to the end of the observation period.

 

Acute dermal toxicity

1) Study with test substance

Four rabbits per dose group(doses unknown) were treated with the test substance under standardized conditions at dose levels not specified, and were observed for 14 days (Smyth et al. 1969; Rel 2). The reported acute dermal LD50 determined according to the Thompson's method was approx. 452.5 mg/kg bw (280.6 – 742.1 mg/kg bw). No further details were available.

 

2) Study with salt of the test substance

A salt of the test substance was recently examined in an additional acute dermal toxicity study conducted according to the OECD guideline 402 (Val 1; BASF SE, 2009), in which young adult Wistar rats (5 males and 5 females) were dermally exposed to two single doses of 3420 and 1710 mg/kg bw of methyl-diamino-cyclohexan dihydrochlorid (as solution in doubly distilled water) applied to the clipped skin (dorsal and dorso-lateral parts of the trunk) and under semi-occlusive dressing for 24 hours. The animals were then observed for 14 days.

Because no mortality occurred and no clinico-pathological or local effects were observed, the acute dermal median lethal dose (LD50) was determined to be > 3420 mg/kg bw.

 

3) Interpretation of acute dermal toxicity

It is postulated that the low LD50 obtained in rabbits (Smyth et al., 1969) is a consequence of a susceptible reaction of test animals to stress, e.g. caused by local skin damage, since emotional stress and intense pain can lead to cardiac failure in rabbits. A distinction between stress-induced and systemic toxicity-induced lethality by the test substance is therefore relatively impossible. This assertion is strengthened by the fact that no systemic toxicity was observed in rats after acute dermal treatment (BASF SE, 2009), although it can be anticipated that the test substance would become systemically available after dermal absorption of its salt. Therefore, the assessment of the acute dermal toxicity of the registered substance will solely be based on the acute study in Wistar rats which is one of the recommended test species according to OECD TG 402.

Justification for classification or non-classification

No guideline conformed studies were available for the evaluation of the acute oral toxicity of the registered substance. The only available indications were obtained from a poorly documented publication (Smyth et al., 1969), in which the acute LD50 was approx. 1276.1 mg/kg bw for the oral route. In the same study, no deaths occurred when test animals were exposed to an atmosphere saturated with vapours of the test substance for 8 hours. A dermal acute LD50 of approx. 452.5 mg/kg bw was found in rabbits (Smyth et al., 1969), probably as a consequence of local damage, but was >3420 mg/kg bw in rats treated with test substance salt according to the OECD TG 402.

Due to the corrosivity of the test substance, further acute toxicity studies with the registered substance does not have to be conducted according to annex VIII of the REACH Regulation (Standard Information Requirements for Substance Manufactured or Imported in Quantities of 10 Tonnes or More; Toxicological Information; Column 2).

Classification,Labelling, and Packaging Regulation (EC) No. 1272/2008

The available experimental test data are reliable and suitable for classification purposes under Regulation 1272/2008. As a result the substance is considered to be classified for acute oral toxicity (cat. 4, H302) under Regulation (EC) No. 1272/2008, as amended for the third time in Directive (EC 618/2012).