Bis(2-dimethylaminoethyl)(methyl)amine

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Comparable to guideline study with acceptable restrictions (no details on test substance, e.g. purity, no data on individual animals availabvle; limited results description - body weight data missing, DPM values not available)

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1997

Materials and methods

Test guideline

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)

GLP compliance:

yes

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

Bis(2-dimethylaminoethyl)(methyl)amine

EC Number:

221-201-1

EC Name:

Bis(2-dimethylaminoethyl)(methyl)amine

Cas Number:

3030-47-5

Molecular formula:

C9H23N3

IUPAC Name:

(2-{[2-(dimethylamino)ethyl](methyl)amino}ethyl)dimethylamine

Test material form:

not specified

Details on test material:

Test material (CAS #3030-47-5) obtained from Air Products and Chemicals, Lot #AW411814M4

In vivo test system

Test animals

Species:

mouse

Strain:

Balb/c

Sex:

not specified

Details on test animals and environmental conditions:

Supplier of animals: Charles River
The age of the mice ranged from 9 to 11 weeks.
All animals were observed cageside at least twice daily for morbidity, mortality, availability of food and water and treatment effects.
These observations included evaluation of the skin, fur and mucous membranes, respiration, nervous system and behavior patterns.

The animal rooms of the testing facility were designed to maintain adequate environmental conditions concerning temperature, humidity, and
photocycle and are regulated for the species under test. The temperature was set at 22 +/- 1 °C, the humidity at 50 +/- 10% RH and the
lightldark cycle is set at 12 hours on: 12 hours off.
Feed analysis was performed by Purina Mills, Inc. and analysis of the tap water (municipal water supply) was performed in
accordance with Laboratory Standard Operating Procedures.

Study design: in vivo (LLNA)

Vehicle:

acetone/olive oil (4:1 v/v)

Concentration:

1%

No. of animals per dose:

5

Details on study design:

Study consisted of a vehicle control group (5 mice treated with a solution of 4:l acetone:olive oil, AOO), three positive control
groups (5 mice treated with a solution of each 1.0% dinitrochlorobenzene (DNCB), 0.17% benzalkonium chloride (BC), and 25% trimellitic
anhydride (TMA), and treatment groups (5 mice treated with solutions of the test substance in AOO). The test substance was tested in
the LLNA at the minimal irritating concentration (MIC) determined in previous Primary Dermal Irritation Studies (PDIS).

Prior to the application of the solutions on Day 1, an erythema score was assessed for each mouse. The solutions were applied by
dispensing 12.5 ul of the appropriate solution onto the dorsal and ventral side of each ear - each ear received a total of 25 ul of solution for a total of 50 ul per mouse. The solutions were applied once daily for three consecutive days. On Day 5, the erythema score was assessed for each mouse. Each mouse was then injected, via the lateral tail vein, with 0.2 mi of phosphate buffered saline (PBS)
containing 20 uCi of 3H-thymidine. Five hours after the injections, the mice were sacrificed by C02 inhalation and the draining auricular
lymph nodes were excised and pooled for each mouse. A single cell suspension of lymph node cells (LNC) was prepared and
3H-thymidine incorporation was measured on a beta-scintillation counter. Radioisotope incorporation was measured as disintegrations per minute (dpm) per mouse and a mean dpm value +/- SE (standard error) was calculated for each experimental group.
In addition, a stimulation index (SI) was calculated using absolute dpm value for each mouse as the numerator, and the mean dpm value
from the vehicle control mice as the denominator. A mean SI k SE was calculated for each experimental group. Any chemical that produces a SI of > 3 in the LLNA was considered "positive".

Statistics:

Calculations were made using Microsoft Excel. The following equations were used:
1. SI = dpm of mouse / mean dpm of vehicle group
2. AVERAGE* = "average" of numbers
3. STDEV* = "estimates standard deviation of a population based on a sample"
4. Standard Error (SE) = STDEV s the square root of the number of mice in group
* Defined by Microsoft Excel

Results and discussion

Positive control results:

Trial 1:
TMA, but not DNCB produced a robust response. DNCB failed to trigger a SI well in excess of 3.0 in Trial 1.
BC triggered a slight SI, which was less than 3.0 and consistent with the fact that it is an irritant; but not a sensitizer.
Trial 2:
SI of 2.80 for 0.17% BC, SI of 46.10 for 1.0% DNCB, and SI of 44.76 for 25% TMA.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: A mean SI +/- SE was calculated for the test material from two separate trials Trial 1: SI: 3.53 +/- 0.50 Trial 2: SI: 1.61 +/- 0.32 Overall mean: SI: 2.57 +/- 0.42.

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: data not available

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Remarks:

Migrated information