1-Propanaminium, N-(carboxymethyl)-3-(formylamino)-N,N-dimethyl-, inner salt

Endpoint summary

Key value for chemical safety assessment

Skin sensitisation

Link to relevant study records

Reference

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

15. Nov. 2004 - 17. Dec. 2004

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.6 (Skin Sensitisation)

Deviations:

no

GLP compliance:

yes

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

A valid GPMT conducted according to guideline is available, which is reliable without restrictions and adequate for classification and labelling purposes. Potency estimation is not mandatory when existing guideline and GLP conforming data are available, which were conducted before the new annex of the REACH Regulation entered into force. Moreover, no indication for skin sensitisation was observed in this study, thus, no dose response information is needed. For this reason and for reasons of animal welfare no additional LLNA was conducted.

Species:

guinea pig

Strain:

other: Pirbright White, BOR DHPW

Sex:

female

Details on test animals and environmental conditions:

Species: guinea pig
Breed: Pirbright White, BOR DHPW
Breeder: Harlan Winkelmann, Borchen, Germany
Date of receipt: 2004-11-03
Sex: female
Number: 20 (test group 10, control group 5, pilot experiment 5)
Body weights: 313.3 g - 415.6 g
Age: - 4 weeks on delivery
Identification: dye marks and cage numbers
Diet: Altromin International, Lage, Germany, 3022 - 1420
Water: Drinking water for the animals consisted of normal tap water from municipal sources: Stadtische Werke Krefeld AG, Abt. 2 TGW, 47804 Krefeld, Germany. The composition of the water is regularly determined. The data are retained in the archive. The water was supplied to the animals ad libitum via drinking bottles with rubber stoppers and steel pipes.
Bedding Material: Lignocel BK 10/20, Rettenmaier, Jagstzell, Germany
Husbandry: The animals were housed with 2 or 3 animals in Makrolon®-cages No.4. A non-barrier system with air conditioning was used. The air conditioning had the following nominal values: Temperature: 22°C ± 3°C, Humidity: 30 % - 70 %, Air exchange: 8 times/hour. Climate control was run automatically. The lightening was in a 12-hour light/dark-cycle. Temperature and humidity were recorded continuously using a thermohygrometer, Lambrecht GmbH, Gbttingen, Germany. In cause of a technical default of the thermohygrometer the climate could not be recorded on the 2nd, 3rd, 4th and 5th of December 2004. The humidity was lower then 30 % from the 6th to 13th December 2004. These deviations are not supposed to have any significant effect on the experimental result.

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Concentration / amount:

Pilot experiment:
Intradermal: 0.1 ml of 5.0 %, 3.5 %, 2.0 % and 0.5 % (w/w) dilution in saline (0.9 % NaCI solution).
Dermal: soaked patch with 100 %, 75 %, 50 % and 25 % (w/w) of the test substance in saline.
Challenge: duhring chamber with 50 % and 30 (w/w) of the test substance in saline.
Main study:
Intradermal: 0.1 ml of 3.5 % (w/w) solution of the test substance in saline.
Dermal: soaked patch with the undiluted test substance.
Challenge: duhring chamber with 30% (w/w) solution of the test substance in saline.

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Concentration / amount:

Pilot experiment:
Intradermal: 0.1 ml of 5.0 %, 3.5 %, 2.0 % and 0.5 % (w/w) dilution in saline (0.9 % NaCI solution).
Dermal: soaked patch with 100 %, 75 %, 50 % and 25 % (w/w) of the test substance in saline.
Challenge: duhring chamber with 50 % and 30 (w/w) of the test substance in saline.
Main study:
Intradermal: 0.1 ml of 3.5 % (w/w) solution of the test substance in saline.
Dermal: soaked patch with the undiluted test substance.
Challenge: duhring chamber with 30% (w/w) solution of the test substance in saline.

No. of animals per dose:

main study: test group 10 animals, control group 5 animals.

Details on study design:

RANGE FINDING TESTS: Pilot Experiment:
The purpose of the pilot experiment was to determine which concentration of the test substance 1. led to slight irritation after intradermal applicatio (determination of the maximum compatible dose), 2. led to slight irritation after dermal application and 3. can just be applied dermally without leading to skin irritation (subirritative dose).

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 2 (intradermal and dermal)
- Exposure period: Dermal induction (for 48 h) 7 days after induction by Intradermal injection
- Test groups: 1 with 10 animals
- Control group: 1 with 5 animals
- Frequency of applications: once
- Concentrations: Intradermal: 0.1 ml of 3.5 % (w/w) solution of the test substance in saline. Dermal: soaked patch with the undiluted test substance. Challenge: duhring chamber with 30% (w/w) solution of the test substance in saline.

B. CHALLENGE EXPOSURE
- No. of exposures: 1
- Day(s) of challenge: at day 21 of the application period
- Exposure period: 24 hours
- Test groups: 1 with 10 animals
- Control group: 1 with 5 animals
- Site: left flank
- Concentrations: 30 % (w/w) dilution of the test substance in saline
- Evaluation (hr after challenge): 24 h and 48 h after end of challenge exposure

OTHER:

Positive control substance(s):

yes

Remarks:

The corresponding validation experiment produced a sensitisation rate of 70 % with the reference substance 2-mercaptobenzothiazole showing the sensitivity of the test system.

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

30 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 30 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Reading:

2nd reading

Hours after challenge:

48

Group:

test chemical

Dose level:

30 %

No. with + reactions:

0

Total no. in group:

10

Clinical observations:

none

Remarks on result:

other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 30 %. No with. + reactions: 0.0. Total no. in groups: 10.0. Clinical observations: none.

Reading:

1st reading

Hours after challenge:

24

Group:

negative control

Dose level:

0 %

No. with + reactions:

0

Total no. in group:

5

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

100 %

No. with + reactions:

7

Total no. in group:

10

Remarks on result:

positive indication of skin sensitisation

Challenge results

 

Animal number	Code	Sex	24 hours		48 hours
			Erythema	Edema	Erythema	Edema
Test Group
1	lv	female	0	0	0	0
2	Rv	female	0	0	0	0
3	Lh	female	0	0	0	0
4	Rh	female	0	0	0	0
5	2xv	female	0	0	0	0
6	Lv	female	0	0	0	0
7	rv	female	0	0	0	0
8	Lh	female	0	0	0	0
9	Rh	female	0	0	0	0
10	2xv	female	0	0	0	0
Control Group
1	Lv	female	0	0	0	0
2	Rv	female	0	0	0	0
3	Lh	female	0	0	0	0
4	Rh	female	0	0	0	0
5	2xv	female	0	0	0	0

 

Interpretation of results:

not sensitising

Remarks:

Migrated information Criteria used for interpretation of results: other: CLP, EU GHS (Regulation (EC) No 1272/2008)

Conclusions:

The observed sensitisation rate of 0 % indicates that the test substance Formamidopropyldimethylbetaine is a non-sensitising compound in this test system.

Executive summary:

For labelling and classification purposes the test substance Formamidopropyldimethylbetaine was tested regarding its skin sensitisation potential. The test was performed according to the stringent protocol of Magnusson and Kligman, in which adjuvant is used to elicit an immunologic reaction. The challenge application was executed with Duhring chambers to aim for better, i.e. more exaggerated, exposure conditions. The test protocol is in compliance with the OECD Guideline for the Testing of Chemicals No. 406 Skin Sensitisation (1992). The maximum compatible doses for the intradermal and dermal application as well as the subirritative dose for the challenge were determined in a pilot experiment. In the main experiment ten (10) female guinea pigs of the strain "Pirbright White" were treated intradermally with 0.1 ml of a 3.5 % (w/w) dilution of the test substance in saline. For the dermal induction the undiluted test substance was used. Intradermal application of the 3.5 % (w/w) dilution of the test substance in saline caused slight to moderate erythema and edema formation and the topical application of the undiluted test substance caused slight erythema and edema formation. In the challenge application Duhring chambers with a 30 % (w/w) dilution of the test substance in saline were used, a test concentration which was definitely non-irritating. Besides crust formation at the injection sides after intradermal application with FCA no other

systemic toxic symptoms or toxic reactions were observed at any time during the study. Body weight development of the animals was positive and within normal ranges. In the challenge no visible changes on the skin (no erythema and no edema) were observed at any time point; i.e. there was no positive challenge result in the test and the control group animals. Thus, the sensitisation rate was 0 %. Therefore, the test substance Formamidopropyldimethylbetaine has to be regarded as non-sensitising under the applied test conditions when exposed to the skin of experimental animals.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Additional information:

For labelling and classification purposes the test substance Formamidopropyldimethylbetaine was tested regarding its skin sensitisation potential. The test was performed according to the stringent protocol of Magnusson and Kligman, in which adjuvant is used to elicit an immunologic reaction. The challenge application was executed with Duhring chambers to aim for better, L e. more exaggerated, exposure conditions. The test protocol is in compliance with the OECD Guideline for the Testing of Chemicals No. 406 Skin Sensitisation (1992). The maximum compatible doses for the intradermal and dermal application as well as the subirritative dose for the challenge were determined in a pilot experiment. In the main experiment ten (10) female guinea pigs of the strain "Pirbright White" were treated intradermally with 0.1 ml of a 3.5 % (w/w) dilution of the test substance in saline. For the dermal induction the undiluted test substance was used. Intradermal application of the 3.5 % (w/w) dilution of the test substance in saline caused slight to moderate erythema and edema formation and the topical application of the undiluted test substance caused slight erythema and edema formation. In the challenge application Duhring chambers with a 30 % (w/w) dilution of the test substance in saline were used, a test concentration which was definitely non-irritating. Besides crust formation at the injection sides after intradermal application with FCA no other systemic toxic symptoms or toxic reactions were observed at any time during the study. Body weight development of the animals was positive and within normal ranges. In the challenge no visible changes on the skin (no erythema and no edema) were observed at any time point; i.e. there was no positive challenge result in the test and the control group animals. Thus, the sensitisation rate was 0 %. Therefore, the test substance Formamidopropyldimethylbetaine has to be regarded as non-sensitising under the applied test conditions when exposed to the skin of experimental animals.

Migrated from Short description of key information:
In the GPMT test with the test substance Formamidopropyldimethylbetaine, the sensitisation rate was 0 %

Justification for selection of skin sensitisation endpoint:
Data from a GLP compliant guideline study with reliability 1.

Respiratory sensitisation

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not sensitising)

Justification for classification or non-classification

In conclusion, the result of a Magnusson and Kligman study with guinea pigs indicate that Formamidopropyldimethylbetain does not induce skin sensitisation.

According to Directive 67/548/EEC as well as GHS Regulation EC No 1272/2008 Formamidopropyldimethylbetain is not classified as “sensitising” and labelling is not required.