Reaction mass of 1-Propanaminium, 2,3-dihydroxy-N,N-dimethyl-N-[3-[(1-oxododecyl)amino]propyl]-, chloride and 1-Propanaminium, 2,3-dihydroxy-N,N-dimethyl-N-[3-[(1-oxotetradecyl)amino]propyl]- , chloride

Skin sensitisation

Administrative data

Endpoint:

skin sensitisation: in vivo (non-LLNA)

Type of information:

experimental study

Adequacy of study:

key study

Study period:

January 23, 1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1990

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

EPA OTS 798.4100 (Skin Sensitisation)

Version / remarks:

40 CFR Part 798.4100

Deviations:

no

GLP compliance:

yes

Type of study:

Buehler test

Justification for non-LLNA method:

The LLNA method was not available yet by the time the study was conducted

Test material

Test material information

Constituent 1

Reference substance name:

Cocamidopropyl PG-dimonium chloride

Cas Number:

136920-08-6

Molecular formula:

C20-30H43-63ClN2O2 range

IUPAC Name:

Cocamidopropyl PG-dimonium chloride

Test material form:

other: aqueous solution

Details on test material:

Cocamidopropyl PG-dimonium chloride
Lot# PC-329-37

In vivo test system

Test animals

Species:

guinea pig

Strain:

not specified

Sex:

male/female

Details on test animals and environmental conditions:

Healthy USDA certifiedguinea pigs were received from Davidson's Mill Farm. 25 guinea pigs (25males, 0 females) were selected for test based on body weight and health status. Control and test animals were from the same shipment.
acclimation period: 12 days
Caging - gang-caged in suspended stainless steel caging with mesh floors
Room temperature 19-21C
Feed: purina guinea pigs pellets ad-libitum
Water: tap water supplied by automatic water system
Identification: a number was allocated to each rabbit on receipt.
Instillation day: the first induction dose was applied on November 29,1989. after a two week rest period the challenge dose was applied on December 13, 1989. There was no need for a re-challenge.

Study design: in vivo (non-LLNA)

Induction

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.4mL

Day(s)/duration:

6 hours of exposure for each 0.4mL dose. Three repeated doses every week

Adequacy of induction:

other: highest concentration that produce no erythemic or edematous response

Challenge

Route:

epicutaneous, occlusive

Vehicle:

unchanged (no vehicle)

Concentration / amount:

0.4mL neat material

Day(s)/duration:

6hr exposure

Adequacy of challenge:

highest non-irritant concentration

No. of animals per dose:

10

Details on study design:

Animal Selection
Male, Hartley strain, albino guinea pigs allowed to adapt to the laboratory for 12 days. 25 guinea pigs were selected for testing on the basis of good health and body weight. Each guinea pig was uniquely identified with an ear tag and study number. The animals weighed 289-396 grams.

Maximum on-irritating dose
prior to the period of induction, the maximum non-irritating concentration of the test material was determined. The dorsal thoracolumbar region of 2 guinea pigs was rid of hair by clipping. The back of each guinea pig was divided into 4 test sites. (2 sites on each side of the midline). The test material was applied neat and was also diluted with distilled water to yield concentrations of 75, 50 and 25%. Two guinea pig each received the same dose concentrations. by applyng 0.5mL of each respective concentration to pre-assigned delineated test sites on each animal. Each site was covered with 2.5cm2 diameter gauze patch secured in place with hypoallergenic adhesive tape. After 6 hrs of exposure the patches were removed. 24 hrs after initial application, readings were made of local reactions. (erythema and edema). From these results the highest non-irritating concentration that produced ni erythemic or edematous response in 2 guinea pigs and was the concentration used for Induction and Challenge.

Induction and challenge
10 guinea pigs were assigned to the tes group and ten guina pigs were assigned to the control group (0.08% DNCB in 95% ethanol). Immediately prior to test initiation and prior to each induction and each challenge dose, the hair was removed from the dorsal thoracolumbar region of each guinea pig by clipping. One Hilltop chamber was placed over the back of each guinea-pig. 0.4mL of test material was placed into each chamber. after dosing, the test and positive control guinea pigs were each respectively divided into subgroups and placed into stainless steel gang cages. The chambers were removed after six hours of exposure. 24 and 48 hrs after each induction application readings were made of local reactions. This process was repeated once each week until a total of three dose applications had been made. 14 days after the third application, a challenge dose was applied to a naive site on the left of each guinea pig and to the five naive control animals. This site was recorded for a sensitization response for erythema and edema at 24 and 48 hrs after challenge according to the system described in the attached report.

Challenge controls:

(0.08% DNCB in 95% ethanol) 10 guinea pigs

Positive control substance(s):

yes

Results and discussion

Positive control results:

Induction phase
Faint to moderate erythema was observed at all positive control sites 24 and 48 hrs after the second and third application. One site exhibited severe eythema with slight eschar

Challenge phase
at 24hrs, all positive control sites had faint, unusually confluent erythame. This condition persisted at 5 sites through 48 hrs and diminished to very faint erythema at the remaining 5 sites.

In vivo (non-LLNA)

Results

Results 1

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

test chemical

Dose level:

0.4 mL neat material

No. with + reactions:

4

Total no. in group:

10

Clinical observations:

Four guinea pigs exhibited very faint non-confluent erythema 24 hrs after challenge. The remaining 6 treated sites were clear. By 48 hrs only 2 sites still had signs of erythema. All others were clear.

Remarks on result:

no indication of skin sensitisation

Results 2

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

positive control

Dose level:

0.4mL undiluted

No. with + reactions:

10

Total no. in group:

10

Clinical observations:

At 24 hrs, all positive control sites had faint, usually confluent erythema. This condition persisted at 5 sites through 48 hours and diminished at the remaining 5 sites

Remarks on result:

positive indication of skin sensitisation

Results 3

Key result

Reading:

1st reading

Hours after challenge:

24

Group:

other: naive control animals

Dose level:

0.4 mL undiluted

No. with + reactions:

3

Total no. in group:

5

Clinical observations:

3 of 5 previously naive sites exhibited very faint, non-confluent erythema 24 and 48 hrs after challenge. The remaining two sites were clear of irritation at both time points.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on these findings, the substance is not considered a contact sensitizer when applied as received, undiluted, once a week over a 3 week induction period.
The positive response to 0.08% DNCB in 95% Ethyl Alcohol validates the test system used in this study.

Executive summary:

A sample od cocamidopropyl PG-dimonium chloride (>30%) was tested as received, undiluted.

After establishing the highest non-irritating concentration to be 100%, a 3 week induction perdiod was initiated during which 10 young adult, male guinea pigs were treated with the test material and 10 were treated with 0.08% Dinitrochlorobenzene DNCB in 95% ethyl alcohol (positive control). During the induction period the test animals were dosed once a week until a total of three dose applications was achieved. 14 days after the third application a challenge dose was applied to a naive site on each guinea pig and approximately 24 and 48 hrs later the animals were scored for a sensitization response.

A naive control group of 5 animals was maintained under the same environmental conditions and treated with the test material at challenge only.

All guinea pigs appeared active and healthy during the test. There were no signs of gross toxicity, adverse phamacologic effects or abnormal behavior. All animal gained weight.

Induction phase

test animals: during induction, faint to moderate erythema was observed at all test animals sites 24 and 48 hrs after the second and third dose applications

positive control: faint to moderate erythema was observed at all positive control sites 24 and 48 hrs after the second and third dose application. One site exhibited severe erythema with slight eschar.

Challenge phase

test animals: 4 guinea pigs exhibited very faint non confluent erythema 24 hrs after challenge. The remaining 6 treated sites were clear. By 48 hours only 2 sites still had signs of erythema. all other sites were clear.

naive control animals: 3 of5 previously naive exhibited very faint, non-confluent erythema 24 and 48 hrs after challenge. The remaining two sites were clear of irritation at both time points.

positive control animals: at 24 hrs, all positive had faint, usually confluent erythema. This condition persisted at 5 sites through 48 hrs and diminished to very faint erythema at the remaining 5 sites.

The incidence and severity of irritation (sensitization response) after challenge test were as follow:

incidence  incidence  severity  severity    24hrs  48hrs  24hrs  48hrs  test animals  4/10  2/10  0.5  0.5  positive control animals  10/10  10/10  1  0.75  naive animals  3/5  3/5  0.5  0.5  sensitization response

Based on these findings, the substance is not considered a contact sensitizer when applied as received, undiluted, once a week over a 3 week induction period.

The positive response to 0.08% DNCB in 95% Ethyl Alcohol validates the test system used in this study.

positive control