Acacia mearnsi, ext., reaction products with ammonium chloride and formaldehyde

Endpoint summary

Description of key information

The substance Acacia Mearnsi is consider non-irritant when applied on the skin and moderately irritant when applied to the eyes. as a consequence the substance should be classified as Eye Irritant Cat. 2 according to  1272/2008/EC.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 21/07/2006 to 02/08/2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to GLP compliance and International Guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 404 (Acute Dermal Irritation / Corrosion)

Principles of method if other than guideline:

Guideline for testing of chemicals Acute Eye Irritation/Corrosion

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or test system and environmental conditions:

Test system:
Adult white albino rabbits (Oryctolagus cuniculus) of the New Zealand breed, weighing 1850-2200 gramsof live weigh, were used. the animals were healty and had no detectable pathological changes in the skin.
Environmental condition:
The animals were acclimated to the laboratory conditions at least 5 days prior to the beginning of the experiment. They were kept with ventilation of 10 to 15 air changes per room per hour and a photoperiod of 12 hours in the light and 12 hours in the dark. The diet comprised commercial pelletized feed, with supplemetnation of filtered water, both provided ad libitum. 3 animals were used, 3 males, which were kept individually in galvanized wire cages.

Type of coverage:

occlusive

Preparation of test site:

clipped

Vehicle:

unchanged (no vehicle)

Controls:

other: test system used as control

Amount / concentration applied:

The test article Acquapol C1, originally in liquid form, was used pure. The total amount of the solution applied to the skin in each animal was 0.5 ml

Duration of treatment / exposure:

4 hours

Observation period:

14 days

Number of animals:

3

Details on study design:

Approximately 24 hours before the beginning of the test, the hair in the region immediately posterior to the back of the neck were removed using shears for animals, with great care not to hurt and/or irritate the skin. The solution were applied to the small sheared area (approximately 6 cm2) on the back of the animals, and then protected by a gauze/occlusive patch. The untreated adjacent areas served as negative control. At the end of the exposure period (4 hours), the solution residues were removed by rising with water with the aid of cotton. The animals were maintained for 72 hours under observation, being clinically observed. and the lesion grades were noted ast 60 minutes, 24 hours, 48 hours and 72 hours after the application of the product. The animals were examinated for the presence of herythema, edema and formation of eschars, as well as other local or systemic changes.

Analysis of the results:
Each animal was separately evaluated and their results were put into individual table. The degrees of skin lesions were recorded in accordance with the scale in Table 1

Calculation of the skin irritation index:
To estabilish the Skin Irritation Index, the values for erythema and formation of eschars for 24 and 72 hours (2 values) were added to the values for edema formation for 24 and 72 hours (2 values). The total of these 4 values were divide by 2 and classified according to the criterion in table 2.
the edema evaluation was measured through paquimeter, and the calculation of the value of the edema was made through the formula:
(Lat/2) - (Lac/2) = Edmm
Lat= reading of the area tests (entire and with groove)
Lac= reading of the control area (entire and with groove)
Edmm= Value of the edema in millimeters

The graduation of the intensity of the cutaneous reaction was based in the method of Draize, according to the table 1

The calculation of the value of primary cutaneous irritation is accomplished through the toally of the aritmetic average divided by four of the following observations:
- Edema in the entire skin 24:00 hours
-Edema in the skin with groove 24:00 hours
- Eritema in the entire skin 24:00 hours
-Eritema in the skin with groove 24:00 hours
- Edema in the entire skin 24:00 hours
-Edema in the skin with groove 72:00 hours
- Eritema in the entire skin 72:00 hours
-Eritema in the skin with groove 72:00 hours

Classification class are defined in table 2

Irritation parameter:

primary dermal irritation index (PDII)

Basis:

mean

Time point:

other: 24h, 48h, 72h

Score:

ca. 0

Reversibility:

fully reversible

Irritant / corrosive response data:

Skin Irritation Index:
Animal 1: 0, 0
Animal 2: 0, 0
Animal 3: 0,0
Average:0, 0

Classification: Non-irritative Clas IV: No irritation

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

The concerning substance is consider non-irritant when applied on the skin and according to Reg.Ec 1272/2008

Executive summary:

The Primary Skin Irritation Test on rabbits, based on OECD guideline 404, is intended for providing information on the corrosive or irritative effects of a substance on the skin of mammals. The concerning substance showed a skin irritation index 0.0 and is consider non-irritant when applied on the skin in rabbits according to Reg.EC 1272/2008.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 21/07/2006 to 02/08/2006

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Study conducted according to GLP compliance and International Guidelines

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

yes

Remarks:

Observation period was only 7 days long instead 21 days

GLP compliance:

yes

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

Test system:
Adult white albino rabbits (Oryctolagus cuniculus) of the New Zealand breed, weighing 1850-2200 gramsof live weigh, were used. the animals were healty and had no detectable pathological changes in the eyes.
Environmental condition:
The animals were acclimated to the laboratory conditions at least 5 days prior to the beginning of the experiment. They were kept with ventilation of 10 to 15 air changes per room per hour and a photoperiod of 12 hours in the light and 12 hours in the dark. The diet comprised commercial pelletized feed, with supplemetnation of filtered water, both provided ad libitum. 3 animals were used, 3 males, which were kept individually in galvanized wire cages.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent no treatment

Amount / concentration applied:

The test article Acquapol C1, originally in liquid form, was used pure. The total amount of the solution applied to the skin in each animal was 0.1 ml

Duration of treatment / exposure:

24 hours

Observation period (in vivo):

7 days

Number of animals or in vitro replicates:

3

Details on study design:

Approximately 24 hours before the beginning of the test, the animals' eyes to be tested were examinated for the detection of any undesirable change that might exists and render the animal useless for the test. The total volume of the solution was installed onto the conjunctival sac of the left eye, after midly separating both eyelids and subsequently keeping them together for a few seconds.The untreated right eyes was used as negative control.
In the end of the treatment period (after 24 hours), the residue from the applied solution was removed with a physiological solution. The animals so treated were maintained for a maximum period of 7 days following the application and were observed at 1, 24, 48, 72 hours and 7 days for the presence of lesions on the cornea, iris and palpebral and bulbar conjunctivae, as well as other severe local and general alterations.

Analysis of the results:
Each animal was separately evaluated and their results were put into individual tables. The degrees of eye lesions were recorded in accordance with the scale in Table 1

Calculation of the eye irritation index:
Cornea: A x B x 5 (Maximum total = 80)
Iris: A x 5 (Maximum toal = 10)
Conjunctivae: (A + B + C) x 2 (Maximum total = 20)

To estabilish the Eye Irritation Index, the values for cornea, iris and conjunctivae at the respective reading time, obtained through the above formulas, were failled. After obtaining the average reading time for the number of rabbits used, the highest index estabilished was used to classify the product according to the criteria in table 2

Irritation parameter:

maximum mean total score (MMTS)

Basis:

mean

Time point:

other: 48 hours

Score:

ca. 27

Reversibility:

not fully reversible within: 7 days

Irritant / corrosive response data:

Eye Irritability Index: For 48 hours: 27.0
Classification: Class II (Moderately Irritating): Opacity of the cornea and persistent iritis for 7 days, moderate hyperemia of conjunctiva.

Annex 1: Degree of eye lesions in the animals for the product ACQUAPOL C1.

Rabbit n.1
Cornea		Iris	Conjunctiva			Time
0	0	0	3	3	2	1 hour
0	0	1	3	3	2	24 hours
0	0	1	3	3	2	48 hours
0	0	1	2	2	2	72 hours
0	0	0	2	2	2	7 days
Rabbit n.2						-
Cornea		Iris	Conjunctiva			Time
0	0	0	3	3	2	1 hour
1	1	1	3	4	3	24 hours
1	1	1	3	4	3	48 hours
0	0	1	3	3	2	72 hours
0	0	0	2	2	2	7 days
Rabbit n.3
Cornea		Iris	Conjunctiva			Time
1	1	0	2	3	3	1 hour
1	1	0	3	4	3	24 hours
1	1	1	3	4	3	48 hours
1	1	1	3	4	3	72 hours
2	2	2	3	4	3	7 days

Interpretation of results:

moderately irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

The substance Acacia Mearnsi (ACQUAPOL C1) was considered moderately irritant when applied to the eyes in rabbits accordin to the regulation 1272/2008/CE.

Executive summary:

A primary Eye Irritation Test, was conducted on rabbits to study the potential damage effects, reversible or not, of Acacia Mearnsi (ACQUAPOL C1), on the eyes of superior animals. The product was used pure and applied to the conjuctival sac of the left eye of the rabbits at a total volume of 0.1 mL. The animals so treated were mantained for 7 days following the application and observed for the presence of lesion on the eyelid and eyeball mucosae, as well as other severe local and general alterations.

An eye irritation Index of 27.0 was observed on a scale of 110.

The product ACQUAPOL C1 was considered moderately irritating when applied to the eyes in rabbits.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

GLP experimental studies, according to OECD guidelines 404 (for skin irritation) and 405 (for eye irritation) scored as Klimisch 1, are selected as key studies. For each category an additional study, based on Draize Test and scored as Klimisch 2, is selected as supporting study. The substance Acacia Mearnsi is consider non-irritant when applied on the skin and moderately irritant when applied to the eyes.

Skin Irritation:

3 New Zeland Albino rabbits were dermally exposed to 0,5 ml of Acacia Mearnsi (as pure substance). The solution were applied to the small sheared area (approximately 6 cm2) on the back of the animals, and then protected by a gauze/occlusive patch. The untreated adjacent areas served as negative control. At the end of the exposure period (4 hours), the solution residues were removed by rising with water with the aid of cotton. The animals were maintained for 72 hours under observation, being clinically observed. and the lesion grades were noted ast 60 minutes, 24 hours, 48 hours and 72 hours after the application of the product. The animals were examinated for the presence of herythema, edema and formation of eschars, as well as other local or systemic changes . in accordance with OECD 404 guideline. No relevant irritation was observed and the study concluded that Acacia Mearnsi does not show any dermal irritation properties

Eye Irritation:

A primary Eye Irritation Test, was conducted on 3 White Albino rabbits to study the potential damage effects, reversible or not, of Acacia Mearnsi on the eyes. The product was used pure and applied to the conjuctival sac of the left eye of the rabbits at a total volume of 0.1 mL. The animals so treated were mantained for 7 days following the application and observed for the presence of lesion on the eyelid and eyeball mucosae, as well as other severe local and general alterations. An eye irritation Index of 27.0 was observed on a scale of 110 according to OECD 405 guidelines and the substance is considered moderately irritating when applied on the eyes. It should be noted that the observation period was only 7 days and not 21 days as preconised in the OCDE guideline 405. The supporting study (Eunice Mayumi Suenaga, 1998) on rabbit show a reversibility of the effect of Acacia mearnsi extract before the end of the observation periode (14 days).

Justification for selection of skin irritation / corrosion endpoint:
The test was performed according to the GLP and international guideline (study scored Klimisch 1).

Justification for selection of eye irritation endpoint:
The test was performed according to the GLP and international guideline (study scored Klimisch 1).

Effects on eye irritation: moderately irritating

Justification for classification or non-classification

Skin irritation: As Acacia Mearnsi did not induce any skin irritation effects, it does not need to be classified according to classification of Reg. 1272/2008/EC

Eye irritation: Due to the irritation Index and the reversibility of the irritation observed after 14 days (Study report from Eunice Mayumi Suenaga, 1998), Acacia Mearnsi is classified as Eye irritant Cat. 2 according to classification criteria of 1272/2008/EC.