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Skin sensitisation

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Administrative data

Endpoint:
skin sensitisation: in vivo (non-LLNA)
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
1984-07-03 to 1984-08-27
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions

Data source

Referenceopen allclose all

Reference Type:
study report
Title:
Unnamed
Year:
1988
Report Date:
1988
Reference Type:
publication
Title:
Ocular and Dermal Toxicity of Guanidine Nitrate
Author:
Korte DW et al
Year:
1993
Bibliographic source:
International Journal of Toxicology, Vol. 12, No. 6, 592-593

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to
Guideline:
other: Health effects test guidelines, August 1982, EPA 560/6-82-001
Qualifier:
equivalent or similar to
Guideline:
OECD Guideline 406 (Skin Sensitisation)
Deviations:
yes
Remarks:
; only 10 animals were used instead of 20, but as in all 10 animals clear negative results were obtained, this deviation is not considered relevant
GLP compliance:
yes
Type of study:
Buehler test
Justification for non-LLNA method:
A valid Buehler test conducted comparable to guideline with acceptable restrictions is available, which is reliable with 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.

Test material

Reference
Name:
Unnamed
Type:
Constituent
Type:
Constituent
Details on test material:
- Name of test material (as cited in study report): Guanidine Nitrate
- Substance type: pure active substance
- Physical state: white crystalline powder
- Analytical purity: 99.99 %
- Lot/batch No.: 123820
- Expiration date of the lot/batch: not available
- Stability under test conditions: stable in acqueous solution (20 μg/ml) at room temperature for
at least 12 days

In vivo test system

Test animals

Species:
guinea pig
Strain:
Hartley
Sex:
male
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Charles River Breeding Laboratories, Wilmington, MA)
- Age at study initiation: 33 days
- Weight at study initiation: 178-225 g at 15 days prior to first induction dose
- Housing: individually in stainless steel, wire mesh cages
- Diet: ad libitum, Certified Purina Guinea Pig Chow Diet 5026 (Ralstone Purina Company, Checkerboard Square, St Louis, MO)
- Water: ad libitum, continuous drip from a central line
- Acclimation period: 15 days before adminisrtation of the first induction dose

ENVIRONMENTAL CONDITIONS
- Temperature: initially 18.9-22.2 °C, increased 22 days after arrival to 20.0-23.3 °C
- Humidity (%): 42-69 % with occasional peaks as high as 88 % during steam line adjustments and room washing
- Air changes (per hr): not specified
- Photoperiod: (hrs dark / hrs light): 12/12

IN-LIFE DATES: From: 1984-07-03 To: 1984-08-27

Study design: in vivo (non-LLNA)

Induction
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
10 % in isotonic saline
Day(s)/duration:
three times at intervals of one week / 6 h exposure
Challenge
Route:
epicutaneous, occlusive
Vehicle:
physiological saline
Concentration / amount:
10 % in isotonic saline
Day(s)/duration:
2 weeks after the last induction / 6 h exposure
No. of animals per dose:
4 - pilot dosing
10 main test treatment
10 main test positive control
10 main test negative control
Details on study design:
PILOT STUDY FOR DOSE RANGE FINDING
4 animals; 100, 10, 1 and 0.1 % concentrations were used to assess the dermal irritation potential

Main study:
INDUCTION
topical application on the left flank
of 0.5ml of a 10 % solution/suspension in isotonic saline
under a 2.5 cm2 gauze patch for 6h;
three times at intervals of one week;
skin evaluation at 24 and 48h following patch removal

CHALLENGE
treatment and positive control animals: two doses on the left and rigth flank
negative control animals: one dose on the left flank
of 0.5ml of a 10 % solution/suspension in isotonic saline
under a 2.5 cm2 gauze patch for 6h;
one time 2 weeks after the last induction;
skin evaluation at 24 and 48h following patch removal

GRADING SYSTEM
Skin reactions were assigned scores according to the following grading system:
0 no reaction; 1 slight erythema; 2 moderate erythema; 3 marked erythema.
Results are expressed in terms of both incidence (the number of animals showing responses of 1 or greater at either 24 or 48h) and severity (the sum of the test scores divided by the number of animals tested).



Positive control substance(s):
yes
Remarks:
2,4 Dinitrochlorobenzene

Results and discussion

Positive control results:
DNCB produced a marked response after the first induction dose, i.e. after the second and third induction and after challenge. Among the positive control animals, the incidence of skin reactions was 9/9 after 24h and 7/9 after 48h. 1/10 animals was lost due to non-compound-related death. The severity of skin reactions was 1.33 after 24h and 1.11 after 48h. Only one response, a 0.5 borderline score, was observed 24h after challenge in one negative control animal. Conclusion: In this study, 24h after challenge treatment performed with 0.1% DNCB, 100% of the animals of the test group were observed with skin reactions.

In vivo (non-LLNA)

Resultsopen allclose all
Key result
Reading:
1st reading
Hours after challenge:
24
Group:
test group
Dose level:
10 % in isotonic saline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: test group. Dose level: 10 % in isotonic saline. No with. + reactions: 0.0. Total no. in groups: 10.0.
Key result
Reading:
2nd reading
Hours after challenge:
48
Group:
test group
Dose level:
10 % in isotonic saline
No. with + reactions:
0
Total no. in group:
10
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: test group. Dose level: 10 % in isotonic saline. No with. + reactions: 0.0. Total no. in groups: 10.0.
Reading:
1st reading
Hours after challenge:
24
Group:
negative control
Dose level:
10 % in isotonic saline
No. with + reactions:
1
Total no. in group:
9
Clinical observations:
one non-compound-related death
Remarks on result:
other: Reading: 1st reading. . Hours after challenge: 24.0. Group: negative control. Dose level: 10 % in isotonic saline. No with. + reactions: 1.0. Total no. in groups: 9.0. Clinical observations: one non-compound-related death .
Reading:
2nd reading
Hours after challenge:
48
Group:
negative control
Dose level:
10 % in isotonic saline
No. with + reactions:
0
Total no. in group:
9
Clinical observations:
one non-compound-related death
Remarks on result:
other: Reading: 2nd reading. . Hours after challenge: 48.0. Group: negative control. Dose level: 10 % in isotonic saline. No with. + reactions: 0.0. Total no. in groups: 9.0. Clinical observations: one non-compound-related death .

Any other information on results incl. tables

PILOT STUDY AND DOSE RANGE FINDING

Concentration for induction and challenge:10%

SKIN EFFECTS IN INDUCTION

No skin reaction was observed after the first, second and third induction in the control and test animals treated with the test item at 10 % in isotonic saline.

SKIN EFFECTS IN CHALLENGE

A very slight skin reaction was observed in 1/9 negative control animals 24 h after challenge, no skin reaction observed in the test animals: Treatment at challenge was performed with the test item at 10 % in isotonic saline.

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
In this study the test substance Guanidine Nitrate is not a dermal sensitiser.
Executive summary:

In a dermal sensitization study according to the Health effects test guidelines, August 1982, EPA 560/6-82-001 with Guanidine Nitrate (a.i. 99.99%) diluted in isotonic saline, male young adult Hartley guinea pigs (10 per group) were tested using the method of Buehler. The positive control material used in the test was 2,4 -Dinitrochlorobenzene (DNCB).

After dermal induction with 10 % solution of test substance, no skin reaction was observed. Challenge treatment was performed with the same concentration of 10 %. None of the animals of the test group showed any skin effects, whereas one animal of the control group showed a very slight skin reaction.

 

The sensitization rate at 24 h and at 48 h was 0 %.

The test article is considered to be a non-sensitizer in this study.