Registration Dossier

Administrative data

Endpoint:
respiratory sensitisation: in vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
1991
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
comparable to guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
1991
Report Date:
1991

Materials and methods

Test guideline
Qualifier:
no guideline available
Principles of method if other than guideline:
The study is equivalent to following methods:
- according KAROL et al. 1995; KAROL et al. 1978; DeCEAURRIZ et al. 1987; BARROW et al, '1977, BOTHAM et al., 1988; BOTHAM et al., 1989
- exposition technical criteria according OECD 403 and corresponding EG-guideline as relevant so far for this study type
- Measuring according to general recommendations of ASTM E 981-84; ALARIE, 1973
GLP compliance:
yes (incl. certificate)

Test material

Reference
Name:
Unnamed
Type:
Constituent
Test material form:
solid: particulate/powder
Details on test material:
Reactive Orange 64

Test animals

Species:
guinea pig
Strain:
Dunkin-Hartley
Remarks:
Pirbtight-White
Sex:
female
Details on test animals and environmental conditions:
TEST ANIMALS
- Source: Versuchstierzucht Winkelmann, Borchen
- Females (if applicable) nulliparous and non-pregnant: yes
- Age at study initiation: 1-2 months
- Weight at study initiation: 235 to 250 g
- Housing: Makrolon-cages type IV (4 animals per cage); cages were cleaned once a week and disinfected with Zephirol (10%), Litter: wooden granular (Typ S 8/15; Ssniff - Spezialdiaten GmbH, D 4770 Soest)
- Diet (e.g. ad libitum): ad libitum; fixed formula standard diet “Altromin 3022 – Haltungsdiät für Meerschweinchen from Altromin GmbH, Lage)
- Water (e.g. ad libitum): ad libitum; tap water
- Acclimation period: at least 1 week
- Indication of any skin lesions: only health and symptom free animals were used for the study.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 22 ± 2 °C
- Humidity (%): ca. 50%
- Air changes (per hr): 10
- Photoperiod (hrs dark / hrs light): 12 hours light/12 hours dark

In-life dates:
Induktionsphase: 13.05.91-- 17.05.91
Challengephase: 05.0691 - 06.06.91
Sektion: 07.06.91

Test system

Route of induction exposure:
intradermal
Route of challenge exposure:
inhalation
Vehicle:
physiological saline
Concentration:
intradermal induction: 15%
No. of animals per dose:
8
Details on study design:
RANGE FINDING TESTS:

MAIN STUDY
A. INDUCTION EXPOSURE
- No. of exposures: 3 times intradermal application, 2 applications of 100 µL each
- Test groups: 2 applications of 100 µL of 5% test item in peanut oil, each
- Control group: 2 applications of 100 µL of peanut oil, each
- Site: on the flank, Day 0: cranial, Day 2: thoracal, Day 4: caudal
- Frequency of applications: every second day
- Duration: 4 days
- Concentrations: 15% in physiological salin

B. CHALLENGE EXPOSURE
- No. of exposures: 2 à 30 min
- Day(s) of challenge: 2
- Exposure period: day 21 to 24
- Test groups: day 24: 4 mg/m³
- Control group: day 23: 5 or 8 mg/m³
- Site: head-nose exposition
- Concentrations: ca 4 mg/m³ air
- Evaluation (hr after challenge): respiratory rate, respiratory volume, respiratory minute volume, inspiration and expiration time was evaluated during and 20 h after challenge exposure; animals were killed 1 day after the last challenge exposure
Positive control substance(s):
not specified
Negative control substance(s):
other: vehicle

Results and discussion

Results:
Induction:
No test item-related effects other than stainingof the skin

Challenge:
No effects other than lachrymation in some animals

Lung function tests:
Single animals showed changes in respiration. There were no delayed pulmonary reactions. See also table 2 in "Any other information on results incl. tables".

Body weight:
There were no changes in body weights observed.

Gross pathology:
in some animals the lung was bloated

Lung weights:
Lungs of treated animal showed higher weights
Positive control results:
NA
Negative control results:
No effects were observed in vehicle control animals.

Any other information on results incl. tables

Number of Animals with positive Reaction during Challenge Phase

Induction Anaphylaxie Immediate Delayed
Vehicle 0/8 (3)/8 1/8
Test item 0/8 5/8 0/8

Applicant's summary and conclusion

Interpretation of results:
study cannot be used for classification
Conclusions:
No clear test-substance related effects were noted, as some respiratory/sensitising effects were also seen in the vehicle group.
Executive summary:

In a respiratory sensitization study with the test item, young adult Pirbright-White-Dunkin-Hartley guinea pigs were tested for respiratory sensitisation. The test was carried out according to published methods of KAROL et al. 1985 and 1978; DeCEAURRIZ et al. 1987; BARROW et al, 1977, BOTHAM et al., 1988 and BOTHAM et al., 1989.

No effects in body weights were observed compared to the vehicle control. Slightly higher lung weights were seen in test item treated animals. Gross pathological findings in the lung consisted of bloated lungs in some test item treated animals..

During and after the challenge phase, 3/8 control and 5/8 test animals showed reactions of the immediate-type. In addition, one control animal showed delayed effects.

Number of Animals with positive Reaction during Challenge Phase

Induction Anaphylaxie Immediate Delayed
Vehicle 0/8 (3)/8 1/8
Test item 0/8 5/8 0/8

 

Due to the positive effects in vehicle control animals, no clear conclusion on respiratory sensitisation effects can be drawn