Dimethyl(octyl)amine

Endpoint summary

Description of key information

An OECD 404 study was performed in rabbits in order to determine the irritation properties of AD 1 formerly known as Farmin DM08P when applied undiluted to the skin. Moderate erythema and oedema were observed on the test sites of all animals one hour after bandage removal. Severe erythema extending beyond the test site, and very slight to severe oedema were seen from the 24 hour examination to Day 13. Loss of elasticity, eschar formation, darkened skin and a solid test site were also apparent during this time. At termination on Day 16, the test site of one animal was solid, eschar formation was seen on another animal and eschar formation, loss of elasticity and exfoliation was seen on the third animal.

Because of the significant reactions seen at the end of the observation period, the animals were killed following this examination and the test sites were excised and examined in cross section. This examination revealed thickened skin in both animals, but not full thickness destruction of the skin. The control sites did not show any response to the control procedure. The mean values for erythema and oedema recorded 24, 48 and 72 hours after treatment exceeded the EEC limit values considered to indicate a significant inflammatory response to treatment. However, macroscopic examination of the test sites did not reveal full thickness destruction.

On the basis of the findings in this study with no evidence of recovery, according to Regulation EC No. 1272/2008, the test substance AD 1, is classified as Skin Corrosive Category 1B; H314 Causes severe skin and eye damage.

For the investigation of eye irritation an in vitro OECD 437 Bovine Corneal Opacity and Permeability (BCOP) study was performed. The mean in vitro score in this study was 19.32.

An in vivo study similar to guideline OECD 405 but with 6 animals was also available to support the investigation of eye irritation in the rabbit. In this test ocular reactions were recorded 1, 2 and 3 days after treatment according to the Draize scoring system. An assessment of time to recovery was not undertaken. The results of this study were corneal damage in 1/6 animals on Day 2 which was clear by Day 3 but no iritis was reported. Some degree of conjunctival redness and/or chemosis was noted in all animals up to the last observation on Day 3.

It may therefore be concluded on the basis of the findings of the in vitro BCOP study and the in vivo eye irritation study, AD 1 does fulfil the criteria according to Regulation EC No. 1272/2008 for Irritating to eyes. Due to the classification for corrosivity to the skin Category 1B, AD 1 will be labelled H314 Causes severe skin burns and eye damage.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Version / remarks:

1992

GLP compliance:

yes

Specific details on test material used for the study:

A consignment of 300 g (net) Famin DM08P, a clear, colourless liquid, was received from the Sponsor on 21 November 1995. The test material was further identified by the Lot No. 1318. It was kept at ambient temperature, in the original container. The identity, strength and purity of the test material received, and its stability under the storage conditions above, were the responsibility of the Sponsor.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

The animals were housed in a lagomorph room within a limited-access building. The room was kept at slight positive pressure relative to the outside and had its own filtered air supply giving at least 10 complete air changes per hour without re-circulation. Target values for temperature and humidity were 18°C (range 15°-23°C) and 55% R.H. (range 40%-70% R.H.), respectively. The achieved values were monitored daily. Electric time-switches operated a lighting cycle of 12 hours of artificial light per day. An emergency generator was available to maintain the electricity supply in the event of a power failure.
Each animal was inspected on arrival, and unsuitable individuals were rejected. Individual bodyweight was recorded on the day of arrival and at least weekly intervals thereafter. All animals were identified by a uniquely numbered eartag. During the acclimatization period, the health status of each animal was monitored and a record kept. This record was consulted before an animal was allocated to study.
On the day before dosing, the dorsum between the limb girdles was clipped (chemical depilatories were not used). Each animal was examined for abnormality or irritation of the dermal test site before allocation to study.

Type of coverage:

semiocclusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

0.5 mL AD-1 at 100% concentration (as received) applied over approximate 3 x 2 cm area

Duration of treatment / exposure:

The dressings were removed after four hours exposure; the treatment sites were gently washed with warm water and dried with paper tissues to remove excess test material adhering to the skin.

Observation period:

Assessment of skin irritation responses at the control and treated test sites. were made 1, 24, 48 and.72 hours after removal of the bandages. Additional observations of persistent effects of treatment were made on Days 7, 10, 13 and 16. Reactions of the test sites were assessed according to the criteria of Draize (1959) below. Reactions not included below were described as appropriate.

Criteria for assessment of skin irritation responses
Erythema and eschar formation Value
No erythema 0
Very slight erythema (barely perceptible) 1
Well-defined erythema 2
Moderate to severe erythema 3
Severe erythema (beet redness) to slight eschar 4
formation (injury in depth)

Oedema formation Value
No oedema 0
Very slight oedema (barely perceptible) 1
Slight oedema (edges of area well-defined by 2
definite raising)
Moderate oedema (raised approximately 1 mm) 3
Severe oedema (raised more than 1 mm and extending 4
beyond the area of exposure)

Number of animals:

3 (sex not specified)

Details on study design:

Each rabbit was securely restrained by a technician. Two test areas (6 x 6 cm) were marked on either side of the clipped area of dorsum. A · single dose was applied directly to the skin (3 x 2 cm) and covered by an unmedicated gauze patch (3 x 2 cm) which was held in place on the left test site by strips of Blendenn (Community Care Products, 3M Health Care, Loughborough, England). The right test site, acting as a control, was covered by a similar semi-occlusive dressing but otherwise remained untreated. Pads of cotton wool and elasticated bandage were used to protect the patches and ensure good contact between the skin and the test material during the four-hour exposure period. The elasticated bandage was held in place by thin strips of waterproof plaster ('Blenderm') at both edges.

Irritation parameter:

erythema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 16 days.

Remarks:

At termination on Day 16, the test site of the animal has suffered a loss of elasticity, eschar formation and exfoliation was observed.

Remarks on result:

positive indication of irritation

Irritation parameter:

erythema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 16 days

Remarks:

At termination on Day 16, the test site of the animal has suffered eschar formation.

Remarks on result:

positive indication of irritation

Irritation parameter:

erythema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

4

Reversibility:

not fully reversible within: 16 days

Remarks:

At termination on Day 16, the test site of the animal has suffered a loss of elasticity, eschar formation and become solid. Erythema extended beyond test site was observed.

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

animal #1

Time point:

24/48/72 h

Score:

1.3

Reversibility:

fully reversible within: 16 days

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1.7

Reversibility:

fully reversible within: 16 days

Remarks on result:

positive indication of irritation

Irritation parameter:

edema score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Reversibility:

fully reversible within: 16 days

Remarks on result:

positive indication of irritation

Irritant / corrosive response data:

Moderate erythema and oedema were observed on the test sites of all animals one hour after bandage removal. Severe erythema, extending beyond the test site, and very slight to severe oedema were seen from the 24 hour examination to Day 13; loss of elasticity, eschar fonnation, darkened skin and a solid test site were also apparent during this time. At termination on Day 16, the test site of one animal was solid,
eschar fonnation was seen on another rabbit and eschar formation, loss of elasticity and exfoliation were seen on the third animal.
Because of the significant reactions seen at the end of the observation period, the animals were killed following this examination and the test sites were excised and examined in cross section. This examination revealed thickened skin in both animals, but not full thickness destruction of the skin.
The control sites did not show any response to the control procedure.
The mean values for erythema and oedema recorded 24, 48 and 72 hours after treatment exceeded the EEC limit values considered to indicate a significant inflammatory response to treatment. However, macroscopic examination of the test sites did not reveal full thickness destruction and the test material was not, therefore, considered to be corrosive.

Interpretation of results:

Category 1B (corrosive) based on GHS criteria

Conclusions:

Under the conditions of this test and the criteria of the EEC, Farmin DM08P was classified as 'corrosive' to skin (Category 1B based on GHS criteria).

Executive summary:

The potential of Farmin DM08P to cause inflammatory or corrosive changes upon first contact with skin was assessed by semi-occluded application of 0.5 ml of the test material to the closely-clipped dorsa of three New Zealand White rabbits for four hours. Denna) reactions were assessed 1, 24, 48 and 72 hours after removal of the dressings and on Days 7, 10, 13 and 16.

Moderate erythema and oedema were observed on the test sites of all animals one hour after bandage removal. Severe erythema, extending beyond the test site, and very slight to severe oedema were seen from the 24 hour examination to Day 13; loss of elasticity, eschar formation, darkened skin and a solid test ·site were also apparent during this time. At termination on Day 16, the test site of one animal was solid, eschar formation was seen on another rabbit and eschar formation, loss of elasticity and exfoliation were seen on the third animal. Because of the significant reactions seen at the end of the observation period, two of the animals were killed following this examination and the test sites were excised and examined in cross section. This examination revealed thickened skin in both animals, but not full thickness destruction of the skin.

Under the conditions of this test and the criteria of the EEC, Farmin DM08P was classified as 'corrosive' to skin (Category 1B based on GHS criteria).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2015-11-11 to 2016-01-07

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

other: OECD Guidline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying Ocular Corrosives and Severe Irritants), adopted: 7 September 2009

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Bayerisches Landesamt für Gesundheit und dLebensmittelsicherheit, München, Germany)

Vehicle:

unchanged (no vehicle)

Amount / concentration applied:

The test item was used as provided by the sponsor.

Duration of treatment / exposure:

750 µL of the test substance or the control substance was introduced into the anterior chamber. After 10 minutes incubation at 32+/- 1 °C
either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red).
Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red).
The anterior chamber was refilled with complete RPMI and an illuminance measurement was performed after 2 hours incubation at 32 +/- 1 °C.

Details on study design:

Preparation of the Corneas
The assay uses isolated corneas obtained as a by-product from animals freshly slaughtered at the abattoir A. Moksel AG, Buchloe, Germany.
On the test day, fresh eyes were collected from the slaughterhouse and were transported in HBSS containing Pen/Strep on ice to the
laboratories. Immediately after arrival of the eyes, cornea preparation was initiated.
The eyes were carefully examined for defects and any defective eyes were discarded.
The tissue surrounding the eyeball was carefully pulled away and the cornea was excised leaving a 2 to 3 mm rim of sclera. The isolated corneas
were stored in a petri dish containing HBSS. Before the corneas were mounted in corneal holders (Duratec GmbH) with the endothelial side
against the O-ring of the posterior chamber, they had been visually examined for defects and any defective cornea had been discarded.
The anterior chamber was then positioned on top of the cornea and tightened with screws. The chambers of the corneal holder were then
filled with RPMI (without phenol red) containing 1% FBS and 2 mM L-glutamine (complete RPMI). The posterior chamber was always filled first.
The corneas were incubated for one hour at 32 +/-1 °C.

Calibration of the Opacitometer
The opacitometer (BASF-OP3.0, Duratec GmbH) was switched on at least 15 min before starting the calibration procedure. The filter holder was
placed into the opacitometer and the readout was adjusted to 1000 lux ± 10 lux using the “Calibrate”-turning knob. For calibration the glass
filter F2 was introduced into the filter holder. The readout lied in the range between 540-560 lux. To test the linearity of the measurement,
two additional calibration filters, glass filter F3 and glass filter F4, were measured. For these glass filters, the opacitometer displayed values
between 300-310 lux and between 95-105 lux. The calibration procedure was performed before the test and is documented in the raw data.

Treatment of the Corneas
After the equilibration period, the medium was removed from both chambers and replaced with fresh complete RPMI. An initial measurement was
performed on each of the corneas using the opacitometer. Three corneas with illuminance readings approximately equivalent to the median
illuminance of all corneas were selected as negative-control corneas. The illuminance of each cornea was read and recorded. Only corneas that
had an initial illuminance reading I > I0/1.1651 lux were used for the assay. The medium was removed from the anterior chamber and replaced
with the test item or control.
750 µL of the test substance or the control substance was introduced into the anterior chamber. After 10 minutes incubation at 32 +/- 1 °C
either the test substance or the control substance was removed and the epithelium washed at least three times with MEM (containing phenol red).
Once the medium was free of test substance, the cornea was finally rinsed with complete RPMI (without phenol red). The anterior chamber was
refilled with complete RPMI and an illuminance measurement was performed after 2 hours incubation at 32 +/- 1 °C.
 
After the illuminance measurement was performed, the medium was removed from both chambers of the holder. The posterior chamber was
refilled with fresh complete RPMI. 1 mL of a 4 mg/mL sodium fluorescein solution was added to the anterior chamber and the corneas were
incubated for 90 minutes at 32 +/- 1 °C. Then the medium from the posterior chamber was removed and its optical density at 490 nm (OD490)
was determined, using a spectrophotometer (Jenway 6405 UV/VIS).

Test Groups
3 corneas for the test item
3 corneas as negative controls treated with physiological saline 0.9% NaCl
3 corneas as positive control treated with ethanol 100%
The BCOP assay is considered to be valid if the in vitro irritation score obtained with the positive control falls within the two standard
deviations of the current historical mean.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background
bovine corneas treated with the respective negative control.

Evaluation of Results
The following formula was used to calculate the opacity, whereas the values a and b are equipment-specific variables empirically determined
by the manufacturer:

Opacity= (I_0/I-b)/a

with a = 0.025 and b = 0.9894

The value I0 is the illuminance through a holder without cornea, but with windows and liquid. This value is determined by taking the mean for
a set of cornea holders and is re evaluated periodically.
The change in opacity for each cornea was calculated by subtracting the initial opacity reading from the final opacity reading. These values were
corrected by subtracting from each the average change in opacity observed for the negative-control corneas. The mean opacity value for each
treatment was calculated by averaging the corrected opacity values of each cornea for a given treatment.
The mean OD490 for the blank cuvettes was calculated. The mean blank OD490 was subtracted from the OD490 of each cuvette
(corrected OD490). Any dilutions that were made to bring the OD490 values into the linear range of the spectrophotometer (OD490 should be
less than 1.500), were taken into account by multiplying the OD490 value of the dilution by the dilution factor. The final-corrected OD490 of the
test article and the positive control were calculated by subtracting the average-corrected OD490 of the negative-control corneas from the
corrected OD490 value of each treated cornea:
Final-corrected OD490 = (OD490 – mean blank OD490) – average-corrected negative control OD490
 
The mean OD490 value of each treatment group was calculated by averaging the final corrected OD490 values of the treated corneas for
that treatment condition.
The following formula was used to determine the in vitro irritation score (IVIS):
IVIS = mean opacity value + (15 x mean permeability OD490 value)
The IVIS cut-off values for identifying test substances as inducing serious eye damage (UN GHS Category 1) and test substances not
requiring classification for eye irritation or serious eye damage (UN GHS No Category) are given:
Evaluation of the BCOP Assay
IVIS UN GHS
≤ 3 No Category
> 3; ≤ 55 No prediction can be made
> 55 Category 1
An identification of test substances that should be classified as irritating to eyes (UN GHS Category 2 or Category 2A) or test substances
that should be classified as mildly irritating to eyes (UN GHS Category 2B) cannot be made.
For this purpose further testing with another suitable method is required.

Irritation parameter:

in vitro irritation score

Value:

19.32

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The eye irritancy potential of AD 1 was investigated in the bovine corneal opacity and permeability assay.
The test item was tested as provided by the sponsor.
The following mean in vitro irritation score was calculated: 19.32
No prediction can be made regarding the classification of the test substance AD 1 according to the evaluation criteria. Further testing in
another suitable method is required.
The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and
therefore this assay is considered to be valid.
The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background
bovine corneas treated with the respective negative control.

Conclusions:

No prediction can be made regarding the classification of the test substance AD 1 according to the evaluation criteria.
Further testing in another suitable method is required.

Executive summary:

The eye irritancy potential of AD 1 was investigated in the bovine corneal opacity and permeability assay.

 Preparation of the test item: tested as provided by the sponsor

Mean in vitro irritation score: 19.32

Classification:

 

	UN GHS No Category
X	No prediction can be made
	UN GHS Category 1

The in vitro irritation score obtained with the positive control fell within the two standard deviations of the current historical mean and therefore this assay is considered to be valid.

The negative control responses resulted in opacity and permeability values that are less than the established upper limits for background bovine corneas treated with the respective negative control.

Conclusion

No prediction can be made regarding the classification of the test substanceAD 1 according to the evaluation criteria.

Further testing in another suitable method is required.

Reference 2

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

05-02/1980 - 01/03/1980

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

comparable to guideline study with acceptable restrictions

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

Study performed before guideline available.

GLP compliance:

not specified

Specific details on test material used for the study:

ADMA C-8. There is no confirmation of test item purity ADMA C-8 in report.

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

New Zealand White rabbits, approximately 8 weeks old when received, were equilibrated for at least one week in this laboratory. Six apparently healthy rabbits were selected for the test. The animals were identified by cage tags noting the test material, starting date, animal number and sex. In addition, odd numbered animals in each cage were identified with an indelible ear mark.
The animals were housed 2/cage in suspended wire mesh cages (30" x 18" x 18"). Any extraneous material which might produce eye irritat ion was excluded from the area. Fresh Purina rabbit chow and water were freely available. The animal room, reserved exclusively for rabbits on acute tests, was maintained at 20 - 21 C and was kept clean in accordance with the standards of AAALAC of which this laboratory is an approved member.

Vehicle:

unchanged (no vehicle)

Controls:

no

other: untreated eye served as a control.

Amount / concentration applied:

The test material 0.1 ml was placed once in the conjunctival sac of one eye of each of six rabbits . The lids were held together briefly to insure adequate distribution of the test material.
The untreated eye of each rabbit served as a control.

Duration of treatment / exposure:

Test item was left in palce for duration of study.

Observation period (in vivo):

1, 2 and 3 days after dosing.

Duration of post- treatment incubation (in vitro):

n/a

Number of animals or in vitro replicates:

6 New Zealand white rabbits - 3 male/ 3 female

Details on study design:

The test material 0.1 ml was placed once in the conjunctival sac of one eye of each of six rabbits . The lids were held together briefly to insure adequate distribution of the test material.
The untreated eye of each rabbit served as a control.
The general health of the rabbits was monitored during the observation period. The ocular reactions of the cornea, iris, and conjunctiva were graded at 1, 2, and 3 days after dosing. In addition, eyes were examined
on day three using sodium fluorescein. The eyes were graded by the Draize scoring system. The scores were interpreted as defined in 16 CFR 1500.42.

Irritation parameter:

overall irritation score

Basis:

animal #1

Time point:

24 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #2

Time point:

24 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #3

Time point:

24 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #4

Time point:

24 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #5

Time point:

24 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #6

Time point:

24 h

Score:

4

Irritation parameter:

overall irritation score

Basis:

animal #1

Time point:

48 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #2

Time point:

48 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #3

Time point:

48 h

Score:

10

Irritation parameter:

overall irritation score

Basis:

animal #4

Time point:

48 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #5

Time point:

48 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #6

Time point:

48 h

Score:

4

Irritation parameter:

overall irritation score

Basis:

animal #1

Time point:

72 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #2

Time point:

72 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #3

Time point:

72 h

Score:

10

Irritation parameter:

overall irritation score

Basis:

animal #4

Time point:

72 h

Score:

6

Irritation parameter:

overall irritation score

Basis:

animal #5

Time point:

72 h

Score:

8

Irritation parameter:

overall irritation score

Basis:

animal #6

Time point:

72 h

Score:

4

Interpretation of results:

Category 2 (irritating to eyes) based on GHS criteria

Conclusions:

1/6 rabbits had corneal damage on Day 2 which was clear by Day 3, There was no iritis.
Some degree of conjunctival redness and/or chemosis was noted in all animals.

Executive summary:

Eye irritation test of N,N-dimethyloctan-1-amine carried out on one eye of 6 White New Zealand rabbits with the non-treated eye acting as a control.

The test material, 0.1 ml was placed once in the conjunctival sac of one eye of each of six rabbits . The lids were held together briefly to insure adequate distribution of the test material.

The general health of the rabbits was monitored during the observation period. The ocular reactions of the cornea, iris, and conjunctiva were graded at 1, 2, and 3 days after dosing. In addition, eyes were examined on day three using sodium fluorescein. The eyes were graded by the Draize scoring system. The scores were interpreted as defined in 16 CFR 1500.42.

In conclusion, 1/6 rabbits had corneal damage on Day 2 which was clear by Day 3, There was no iritis. Some degree of conjunctival redness and/or chemosis was noted in all animals.

N,N-dimethyloctan-1-amine is classed as Catergory 2 (irritating to eyes).

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available