D-gluconic acid, compound with N,N''-bis(4-chlorophenyl)-3,12-diimino-2,4,11,13-tetraazatetradecanediamidine (2:1)

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2000

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Remarks:

The study was specifically conducted to compare the sensitivity of the two standard skin sensitisation tests which are recommended by the OECD guideline 406. Only 5 animals/group were used, but 5 groups were used which received different concentrations of the test substance increasing the total number of animals and the sensitivity of the assay. There was no positive control in a strict way, but all other tested substances (including known skin sensitizers) in the study (chloraniline, eugenol, formaldehyde, mercaptobenzothiazole and neomycin sulphate) gave positive results demonstrating the validity of the assay. These alterations are not considered to be of any concern and the study is regarded relevant to assess the sensitisation potential of chlorhexidine digluconate.

Data source

Reference

Reference Type:

publication

Title:

Comparison of the sensitivities of the Buehler test and the guinea pig maximization test for predictive testing of contact allergy

Author:

Frankild S, Volund V, Wahlberg JE & Andersen KE

Year:

2000

Bibliographic source:

Acta Derm Venereol 80, 256-262

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 406 (Skin Sensitisation)

Principles of method if other than guideline:

Pilot study: Yes
The procedure was performed according to Magnusson and Kligman. The GPMT procedure was modified with a multiple-dose design: 30 animals were assigned to one control group of 5 animals and 5 test groups containing 5 animals each, using different induction concentrations of the allergen for each group. Simultaneous increases in both intradermal and topical doses induction doses were used. In the multiple-dose GPMT procedure pretreatment with SDS was omitted.

GLP compliance:

no

Type of study:

guinea pig maximisation test

Justification for non-LLNA method:

Already Existing Study

Test material

Test material information

Constituent 1

Reference substance name:

D-gluconic acid, compound with N,N''-bis(4-chlorophenyl)-3,12-diimino-2,4,11,13-tetraazatetradecanediamidine (2:1)

EC Number:

242-354-0

EC Name:

D-gluconic acid, compound with N,N''-bis(4-chlorophenyl)-3,12-diimino-2,4,11,13-tetraazatetradecanediamidine (2:1)

Cas Number:

18472-51-0

Molecular formula:

C22H30Cl2N10.2C6H12O7

IUPAC Name:

N',N'''''-hexane-1,6-diylbis[N-(4-chlorophenyl)(imidodicarbonimidic diamide)] - D-gluconic acid (1:2)

Details on test material:

no further data

In vivo test system

Test animals

Species:

guinea pig

Strain:

Dunkin-Hartley

Sex:

female

Details on test animals and environmental conditions:

Source: Salblins, Malmö, Sweden
Weight at study initiation: 300-350 g

Study design: in vivo (non-LLNA)

Induction

Induction 1

Route:

intradermal and epicutaneous

Vehicle:

physiological saline

Induction 2

Concentration / amount:

Concentrations used for induction: 0.001-1 % in saline (intradermal); 0.01-10 % in saline (topical); tested up to highest concentration tolerated systemically and not producing skin necrosis
Concentration Freunds Complete Adjuvant (FCA): not reported
Concentrations used for challenge: 0.1 and 0.3 % in saline (up to maximum non-irritant concentration)

Challenge

Challenge 1

Route:

epicutaneous, occlusive

Vehicle:

physiological saline

Challenge 2

Concentration / amount:

Concentrations used for induction: 0.001-1 % in saline (intradermal); 0.01-10 % in saline (topical); tested up to highest concentration tolerated systemically and not producing skin necrosis
Concentration Freunds Complete Adjuvant (FCA): not reported
Concentrations used for challenge: 0.1 and 0.3 % in saline (up to maximum non-irritant concentration)

No. of animals per dose:

5/group (5 treatment groups)

Details on study design:

Induction schedule: day 0 (intradermal + FCA) – day 7 (topical, occlusive)
Challenge schedule: day 21 (challenge) – day 35 (rechallenge)
Rechallenge: Yes
Scoring schedule: 48 h and 72 h after challenge

Challenge controls:

5

Positive control substance(s):

yes

Remarks:

chloraniline, eugenol, formaldehyde, mercaptobenzothiazole and neomycin sulphate

Results and discussion

Positive control results:

The other 5 tested substances gave a positive reaction

In vivo (non-LLNA)

Results

Results 1

Reading:

other: detailed scoring not reported

Group:

test chemical

Dose level:

0.01-10%

No. with + reactions:

0

Total no. in group:

5

Remarks on result:

other: Reading: other: detailed scoring not reported.

Results 2

Group:

negative control

Remarks on result:

other: not reported

Results 3

Group:

positive control

Remarks on result:

other: not reported

Any other information on results incl. tables

Results of pilot studies: Highest non-irritating topical concentration for challenge: 0.3 %.

Challenge: No animals in the test groups and no animals in the control showed a positive response to chlorhexidine digluconate in the challenge.

Re-challenge: Not explicitly stated, but chlorhexidine digluconate was reported to give negative result in the complete test.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Under the conditions of the assay, chlorhexidine digluconate was not sensitising in a GPMT.

Executive summary:

This is a scientific study in accordance with generally accepted scientific principles. The study was specifically conducted to compare the sensitivity of two standard skin sensitisation tests recommended by the OECD guideline 406.

Under the conditions of the assay, chlorhexidine digluconate was not sensitising in a GPMT.