6-aminopenicillanic acid

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

29 December 2009 to 12 January 2010

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: GLP study conducted to current accepted guidelines

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2010

Materials and methods

Test guideline

Test guideline 1

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

Test guideline 2

Qualifier:

according to guideline

Guideline:

EU Method B.42 (Skin Sensitisation: Local Lymph Node Assay)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

With the exception of no analysis carried out to determine the homogeneity, concentration or stability of the test material formulation, the work was performed in compliance with UK GLP standards (Schedule 1, Good Laboratory Practice Regulations 1999)

Type of study:

mouse local lymph node assay (LLNA)

Test material

Test material information

Constituent 1

Reference substance name:

6-aminopenicillanic acid

EC Number:

208-993-4

EC Name:

6-aminopenicillanic acid

Cas Number:

551-16-6

Molecular formula:

C8H12N2O3S

IUPAC Name:

6-amino-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid

Details on test material:

- Name of test material : 6-Aminopenicillanic acid
- Substance type: Off white powder
- Physical state: Solid
- Analytical purity: 98.7 % (w/w)
- Impurities : Any other single impurity 0.07 %, Penicinic acid 0.17 %, Related Substance and Degradation Products 0.85 %
- Purity test date: 29 May 2009
- Lot/batch No.: 09X10247
- Expiration date of the lot/batch: 26 May 2011
- Storage condition of test material: Room temperature in the dark

In vivo test system

Test animals

Species:

mouse

Strain:

CBA

Sex:

female

Details on test animals and environmental conditions:

TEST ANIMALS
- Source: Harlan Laboratories UK Limited, Bicester, Oxon, UK
- Age at study initiation: 8 to 12 weeks of age
- Weight at study initiation: 15 to 23 g
- Housing: The animals were individually housed in suspended solid floor polypropylene cages furnished with softwood woodflakes. The animals were provided with environmental enrichment items.
- Diet: ad libitum 2014 Teklad Global Rodent diet supplied by Harlan Teklad, Blackthorn, Bicester, Oxon, UK
- Water : Mains tap water ad libitum
- Acclimation period: At least five days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 19 to 25 °C
- Humidity (%): 30-70 %
- Air changes (per hr): 15 per hour
- Photoperiod (hrs dark / hrs light): lighting was controlled by a time switch to give twelve hours continuous light (06.00 to 18.00) and twelve hours darkness.

Study design: in vivo (LLNA)

Vehicle:

dimethyl sulphoxide

Concentration:

5, 15 and 50 % w/w in dimethyl sulphoxide (25 µL)

No. of animals per dose:

5 mice per group

Details on study design:

RANGE FINDING TESTS:
- Compound solubility: DMSO was found to give the highest concentration for dosing.
- Irritation: No irritation or systemic toxicity effects were noted.

MAIN STUDY
ANIMAL ASSIGNMENT AND TREATMENT
- Criteria used to consider a positive response: the test material would be regarded as a sensitiser if at least one concentration of the test material resulted in a threefold or greater increase in 3HTdR incorporation compared to control values. Any test material failing to produce a threefold or greater increase in 3HTdR incorporation would be classified as a "non sensitiser".

TREATMENT PREPARATION AND ADMINISTRATION: The test material was prepared 2 hours prior to administration and mixed by vortexing.

Positive control substance(s):

hexyl cinnamic aldehyde (CAS No 101-86-0)

Statistics:

Data was processed to give group mean values for disintegrations per minute and standard deviations where appropriate. Individual and group mean disintegrations per minute values were assessed for dose response relationships by analysis of homogeneity of variance followed by one way analysis of variance (ANOVA). In the event of a significant result from the ANOVA, pairwise comparisons were performed between control and treated groups. For homogenous datasets Dunnett’s Multiple Comparison test was used and for non homogenous datasets Dunnett’s T3 Multiple Comparison Method was used.

Results and discussion

Positive control results:

The Stimulation Index expressed as the mean radioactive incorporation for the treatment group divided by the mean radioactive incorporation of the vehicle control group is as follows:
15 % v/v α Hexylcinnamaldehyde in dimethyl sulphoxide produced a stimulation index of 6.07 which was classified as a positive response.

In vivo (LLNA)

Results

Results 1

Parameter:

SI

Remarks on result:

other: 5 % 6-APA in DMSO - 0.99 15 % 6-APA in DMSO - 1.03 50 % 6-APA in DMSO - 1.23

Results 2

Parameter:

other: disintegrations per minute (DPM)

Remarks on result:

other: Control (DMSO) - 2244.10 (± 466.54) 5 % 6-APA in DMSO - 2221.80 (± 0.99) 15 % 6-APA in DMSO - 2308.84 (± 591.17) 50 % 6-APA in DMSO - 2750.34 (± 1411.41)

Any other information on results incl. tables

Please find attached the pdf document Appendix I, containing tabulated data on sensitisation and toxicity screening.

Applicant's summary and conclusion

Interpretation of results:

not sensitising

Remarks:

Migrated information

Conclusions:

The test material was considered to be a non sensitiser under the conditions of the test. The test material did not meet the criteria for classification as a sensitiser according to EU labelling regulations Commission Directive 2001/59/EC. No symbol and risk phrase are required.