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Description of key information

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records
Reference
Endpoint:
skin sensitisation: in vivo (LLNA)
Type of information:
experimental study
Adequacy of study:
weight of evidence
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP guideline study
Qualifier:
according to
Guideline:
OECD Guideline 429 (Skin Sensitisation: Local Lymph Node Assay)
GLP compliance:
yes
Type of study:
mouse local lymph node assay (LLNA)
Species:
mouse
Strain:
CBA
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Harlan Winkelmann GmbH, Borchen, Germany
- Age at study initiation: about 6-8 weeks
- Weight at study initiation: 17.3 - 20.0 g
- Housing: individually in Makrolon type I cages
- Diet: ad libitum
- Water: ad libitum
- Acclimation period: 15 days before the first test substance application


ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-24 °C
- Humidity (%): 30-70 %
- Photoperiod (hrs dark / hrs light): 12/12
Vehicle:
other: acetone
Concentration:
0%, 1%, 3% and 10% in acetone
No. of animals per dose:
6
Details on study design:
MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
The animals were assigned randomly to the groups and cages using randomization instructions
- Name of test method: Local Lymph Node Assay
- Criteria used to consider a positive response:
In order to reveal a possible induction of sensitization, the response in the draining lymph
node after epicutaneous application of several concentrations of the test substance to the
skin of the ear backs is determined. The parameters used to characterize the response are
lymph node weight and cell count measured. Because not only sensitization induction
but also irritation of the ear skin by the test substance may induce a lymph node responses, the
weight of ear punches taken from the area of test substance application is determiried as a
parameter for inflammatory ear swelling serving as an indicator for the irritant action of the
test substance.
If a test substance does not show a statistically significant and/or biologically relevant
increase in cell count and/or lymph node weight as compared to the vehicle control in the
presence of statistically significantly and/or biologically relevant increased ear weights as
indication of skin irritation, it is considered not to be a sensitizer .
If at least one concentration tested causes a concentration dependent statistically significant
and/or biologically relevant increase in cell count and/or lymph node weight without being
accompanied by a statistically significant and/or biologically relevant increase in ear weight,
the test substance is considered to be a sensitizer.
If statistically significant and/or biologically relevant increases in ear weights are running in
parallel to the increase in cell count and/or lymph node weight, it cannot be ruled out, that the
lymph node response was caused by irritation and not by skin sensitization. Then, for
identification of the relevance of the statistical evaluation, a comparison of the results of the
present test to appropriate historical control values is performed. If one or a
combination of the measured parameters change statistical significance, evaluation on basis
of the criteria described above may be possible. If the statistical comparison with the
historical control does not yield results useful for evaluation, further investigations may be
necessary to differentiate between irritation and sensitization response.
If a test substance does not elicit a statistical significant increase in lymph node weight and/or
cell count but shows a clear concentration related increase in response, further investigating
of the sensitization potential at higher concentrations should be considered.

TREATMENT PREPARATION AND ADMINISTRATION:
Test substance preparations were produced on a weight by weight (w/w) basis shortly before the appliacation by stirring with a magnetic stirrer.
Vehicle: Acetone
Reason for the vehicle : Acetone was used as the vehicle because good solubility of the preparation was achieved.
Form of application: Solution, epicutaneous application is simulating dermal contact with the compound which is possible to occur under practical use
conditions.
Application volume: 25 μl per ear
Site of application : Dorsal part of both ears
Statistics:
Mean values and standard deviations of the measured parameters were calculated for the test and control groups from the individual values.
The indices of lymph node weight, cell count and ear weight were calculated as the ratio of the test group mean values for these parameters divided by those of the vehicle control group. Lymph node weight, cell count and ear weight were statistically analyzed by Wilcoxon-test.
Parameter:
SI
Remarks on result:
other: see Remark
Remarks:
Treatment of the mice with 1%, 3% and 10% test substance preparations induced statistically significant increase in lymph node cell counts as compared to the vehicle control group. The lymph node weights were statistically increased and therefore in congruency with the cell counts. A concentration < 1% in acetone is considered to be the threshold of sensitization induction and a 1 % concentration in acetone is considered to be the threshold of irritation in the present test model.
Parameter:
other: disintegrations per minute (DPM)
Remarks on result:
other: see Remark
Remarks:
Treatment of the mice with 1%, 3% and 10% test substance preparations induced statistically significant increase in lymph node cell counts as compared to the vehicle control group. The lymph node weights were statistically increased and therefore in congruency with the cell counts. A concentration < 1% in acetone is considered to be the threshold of sensitization induction and a 1 % concentration in acetone is considered to be the threshold of irritation in the present test model.

Treatment

Lymph node weight index

Cell count index

Ear weight index

Vehicle acetone

1.00

1.00

1.00

1 % in acetone

1.56**

2.05**

1.06

3 % in acetone

2.40**

3.02**

1.21**

10 % in acetone

2.57**

3.80**

1.89**

The statistical evaluations were performed using the Wilcoxon-test (* p<0.05; ** p<0.01)

 

 

 

 

No signs of systemic toxicity were noticed. The test substance induced a statistically significant and biologically relevant response of the auricular lymph nodes when applied as 1%, 3% or 10% preparations in acetone. The statistically significant increase in ear weight indicates irritation of the ear skin at concentrations of 3 and 10%, which might have interfered with the lymph node response. The strong increase in ear weight, which was accompanied by crust formation, however, indicated a significant irritant property of the test substance, when applied in these concentrations. The 1% test substance preparation in acetone induced a statistically significant increase in cell count, but only a marginal increase in ear weight. Therefore a concentration < 1% in acetone is considered to be the threshold of sensitization induction and a 1% concentration in acetone is considered to be the threshold of irritation in the present test model.

Thus it is concluded that Tripropylenglykoldiacrylat has a skin sensitizing effect in the Murine Local Lymph Node Assay under the test conditions chosen.

 

 

Interpretation of results:
sensitising
Remarks:
Migrated information
Conclusions:
Tripropylenglykoldiacrylat has a skin sensitizing effect in the Murine Local Lymph Node Assay under the test conditions chosen and is classified in
Annex 1 with R43.
Endpoint conclusion
Endpoint conclusion:
adverse effect observed (sensitising)
Additional information:

There are valid in vivo data available for the assessment of the skin sensitization potential of tripropylene glycol diacrylate.

In a Local Lymph Node Assay, acc. to OECD 429, 6 female CBA mice per dose were treated with 0, 1, 3 and 10 %, of the test material diluted in acetone under standard conditions (BASF AG 2004). 25µL were applied to the dorsum of each ear for three consecutive days. The control group was treated with 25 µl per ear of the vehicle alone. Three days after the last application the mice were sacrificed, and the auricular lymph nodes were removed. The lymph node response was evaluated by measuring the cellular content (indicator of cell proliferation) and weight of each animal's pooled lymph nodes. Moreover, a defined area with a diameter of 0.8 cm was punched out of the apical part of each ear and for each animal the weight of the pooled punches was determined in order to obtain an indication of possible skin irritation. No signs of systemic toxicity were noticed. A statistically significant increase in ear weight as an indication of skin irritation was observed at concentrations of 3 and 10%. A simultaneous response to irritation is indistinguishable from a sensitization response in this test system. The 1% test substance preparation in acetone induced a statistically significant increase in cell count, but only a marginal increase in ear weight. Therefore a concentration < 1% in acetone is considered to be the threshold of sensitization induction and a 1% concentration in acetone is considered to be the treshold of irritation in the present test model.

The application of the test substance at concentrations of 3%, 10% and 30% w/v in acetone resulted in an increase of isotope incorporation of the auricular lymph nodes which was greater than 3-fold at all concentrations.

The sensitizing properties of the test material were also shown in a second LLNA, acc. to OECD 429 (CTL 2001).Groups of four CBA male mice were used for this study. Approximately 25 μl of a 3%, 10% or 30% w/v preparation of the test substance in acetone was applied to the dorsal surface of each ear. A vehicle control group was similarly treated using acetone alone. The procedure was repeated daily for 3 consecutive days. The application of the test substance at concentrations of 3%, 10% and 30% w/v in acetone resulted in an increase in isotope incorporation which was greater than 3-fold at all three concentrations. Following, the third application, the ears of animals dosed with the 30% w/v preparation were red and swollen.

GPMTs were conducted in two further studies (Nethercott 1983, Val. 2). In the first study, Hartley guinea pigs were induced in two stages. In the first stage either side of the shoulder region was injected intradermally. One week after injection the shoulder region of each animal was clipped. The chemical was applied in their respective concentrations (1, 2.5, 5 or 8.5%) in petrolatum over the sites of previous intradermal injection for 48 hours. All animals were challenged two weeks after topical exposure using nonirritant concentrations that had been determined in control animals. Patches were applied to assess irritancy. After 24 hours the patches were removed. The sites were examined at 48 hours for evidence of reaction. Sensitizing effects were seen, at concentrations of 1, 2.5, 5 and 8.5%, 3/10, 4/10, 5/10 and 3/6 animals reacted positively.

In the second GPMT, 15 Dunkin-Hartley guinea pigs (5 controls, 10 substance) were treated with the test material (HRC 1995, Val. 2).The intradermal induction consisted of 0.1 ml of Freund's complete adjuvant 50:50 with sterile water for irrigation, 0.1 ml of TPGDA, 0.25 % v/v in Alembicol D, 0.1 ml of TPGDA, 0.25 % v/v in a 50: 50 mixture of Alembicol D and Freund's complete adjuvant. A volume of 0.1 ml was injected into both the left and right injection sites.

The challenge was a topical application of 30 and 15 % v/v of the test material in Alembicol D two weeks after the topical induction. Also here, positive skin sensitization reactions were observed (8/10, 7/10 and 8/10 positive reactions at 30 % at 1st, 2nd and 3rd reading).

For the sensitising potential of the test substance, a qualitative assessment was conducted:

Although data from local lymphnode assays is available the derivation of an EC3 was not possible. The available LLNA data indicates, that the EC3 is definetly below 1 %.

The available data from the guinea pig tests indicates that the intradermal induction concentration was always above or equal to 0.25 % leading to animals responses between 30 % and 80 % varying from study to study.


Migrated from Short description of key information:
Mouse (LLNA): sensitizing (acc. OECD 429, BASF AG 2004)
Mouse (LLNA): sensitizing (acc. OECD 429, CTL 2001)
Guinea pig (GPMT): sensitizing (Nethercott 1983, Val. 2)
Guinea pig (GPMT): sensitizing (HRC 1995, Val. 2)

Respiratory sensitisation

Endpoint conclusion
Additional information:

No information is available.


Migrated from Short description of key information:
No information is available

Justification for classification or non-classification

Tripropylene glycol diacrylate was demonstrated to be a skin sensitizer. It has also been legally classified as skin sensitizer (Annex I DSD, 67/548/EEC), and is therefore also here classified as R43 according to EU-criteria and as Category 1 according to GHS-criteria.

 

There are no data available to classify tripropylene glycol diacrylate as a sensitizer of the respiratory tract.