Propanoic acid, 2-hydroxy-, ammonium salt (1:1), (2S)-

Eye irritation

Administrative data

Endpoint:

eye irritation: in vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2005-04-06 to 2005-06-17

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2005

Report date:

2005

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Version / remarks:

adopted 24th April 2002

Deviations:

no

GLP compliance:

yes

Test material

Test material information

Constituent 1

Reference substance name:

Propanoic acid, 2-hydroxy-, ammonium salt (1:1), (2S)-

EC Number:

604-012-2

Cas Number:

137296-15-2

Molecular formula:

C3H9O3N

IUPAC Name:

Propanoic acid, 2-hydroxy-, ammonium salt (1:1), (2S)-

Specific details on test material used for the study:

- Name of test material (as cited in study report): Ammonium Lactate, PURASAL® NH
- Physical state: yellowish-white liquid
- pH: 5
- Analytical purity: not reported
- Impurities (identity and concentrations): not reported
- Composition of test material, percentage of components: not reported
- Isomers composition: not reported
- Lot/batch No.: 0503002615
- Expiration date of the lot/batch: 2007-09-28
- Stability under test conditions: not reported
- Storage condition of test material: room temperature (20 ± 5 °C) in the dark

Test animals / tissue source

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: Granja Cunicola San Bernardo, S.L., Tulebras, Navarra, Spain
- Age at study initiation: 14-15 weeks
- Weight at study initiation: 3.1-3.5 kg
- Housing: individually in stainless steel cages (52 × 58 × 43 cm) with a grille floor and placed on a rack
- Diet: free access to a standard diet
- Water: ad libitum
- Acclimation period: at least 7 days

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 20-25
- Humidity (%): 40-70
- Air changes (per hr): not reported
- Photoperiod (hrs dark / hrs light): 12/12

Test system

Vehicle:

unchanged (no vehicle)

Controls:

other: the untreated left eye was used as a control

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 0.1 mL
- Concentration (if solution): undiluted, 100%

Duration of treatment / exposure:

24 h

Observation period (in vivo):

14 days

Number of animals or in vitro replicates:

3

Details on study design:

ADMINISTRATION OF THE TEST ITEM
Both eyes of the animals were examined before treatment in order to reject them if they showed any signs of eye alterations. One single application of the test item was done.The administration was carried out with the animal immobilised and the test item was dropped into the right conjunctival sac, made perfectly accessible by carefully pulling the lower lid outwards. Once the test item had been administered, the lids of the treated eye were held
together for one second. Immediately after administration, the rabbits were fitted with a plastic collar for 1 hour to prevent them from rubbing their eyes.

OBSERVATIONS
The behaviour of the animals was observed immediately following instillation of the product and they were then returned to their cages and observed at regular intervals so as not to miss any serious effect of the test item. The degree of ocular irritation was assessed after approximately 1 hour, and approximately 24, 48, 72 hours, 7 and 14 days after the treatment, following the criteria described below. For the direct examination, the assessments were carried out in the following order: conjunctivae (edema, redness, lacrimation), iris and cornea (degree and extent of opacity). Subsequently, and at all the observations except the one at one hour after administration, the observation of the cornea was completed with the instillation of an aqueous solution of 2 % sodium fluorescein (Colircusi Fluoresceina®, Laboratorios Cusi, S.A., Batch 4CRK1A). This procedure involved the instillation of the aqueous solution of fluorescein in the test area, followed by washing the area with a 0.9 % physiological saline solution. Once the excess fluorescein had been removed, the corneal alterations were observed with the aid of a transilluminator with a cobalt blue filter

SCORING SYSTEM: According to OECD guideline 405

Results and discussion

In vivo

Results

Results 1

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 2

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 3

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 4

Irritation parameter:

iris score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 5

Irritation parameter:

iris score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 6

Irritation parameter:

iris score

Basis:

animal #3

Time point:

24/48/72 h

Score:

0

Max. score:

4

Remarks on result:

no indication of irritation

Results 7

Irritation parameter:

conjunctivae score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.7

Max. score:

4

Reversibility:

fully reversible within: 27 h

Remarks on result:

probability of weak irritation

Results 8

Irritation parameter:

conjunctivae score

Basis:

animal #2

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

probability of weak irritation

Results 9

Irritation parameter:

conjunctivae score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1.7

Max. score:

4

Reversibility:

fully reversible within: 14 d

Remarks on result:

probability of mild irritation

Results 10

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

24/48/72 h

Score:

0.4

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Results 11

Irritation parameter:

chemosis score

Basis:

animal #2

Time point:

24/48/72 h

Score:

0.4

Max. score:

4

Reversibility:

fully reversible within: 48 h

Remarks on result:

probability of weak irritation

Results 12

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

24/48/72 h

Score:

1

Max. score:

4

Reversibility:

fully reversible within: 72 h

Remarks on result:

probability of weak irritation

Irritant / corrosive response data:

The test item was administered initially to one animal (no. 1997). One and 24 hours after the application, some blood vessels hyperaemic (injected) (grade 1) and some swelling above normal (grade 1) was recorded, accompanied by slight lacrimation in the case of the observation carried out 24 hours after the administration. Forty-eight hours after the administration, some hyperaemic blood vessels (injected) (grade 1) were still observed in this animal, but the edema had remitted completely. No ocular lesions were recorded in this animal 72 hours, 7 or 14 days after the administration.
The test item was afterwards applied to two more animals (nos. 2011 and 2012). One hour after the administration, one of these animals (no. 2011) was seen to have redness diffuse, crimson colour, individual vessels not easily discernible (grade 2) accompanied by obvious swelling, with partial eversion of the lids (grade 2). Twentyfour hours afterwards, diffuse redness, crimson colour, and individual vessels not easily discernible (grade 2), were still observed, but the edema had diminished to some swelling above normal (grade 1 ). One hour and 24 hours after the application, slight lacrimation was also observed in this animal. Forty-eight hours after the administration, some hyperaemic blood vessels (injected) (grade 1) were observed. No ocular lesions were recorded 72 hours, 7 or 14 days after the administration.
One, 24 and 48 hours after the administration, diffuse redness, crimson colour, and individual vessels not easily discernible (grade 2) were observed in the remaining animal (no. 2012). One hour after the administration, swelling, with the lids about half closed (grade 3), was also recorded in this animal. This lesion diminished progressively to obvious swelling, with partial eversion of lids (grade 2) during the observation carried out 24 hours after the application, and to some swelling above normal (grade 1) during the observation carried out 48 hours after the application. Slight lacrimation (grade 1) was also recorded in this animal one hour and 24 hours after the treatment. Seventy-two hours and 7 days after the application, some hyperaemic blood vessels (injected) (grade 1) were observed, which had remitted completely 14 days after the administration.

Other effects:

CLINICAL SIGNS AND BODY WEIGHT
The behaviour and physical state of the rabbits, and the body weight evolution were normal throughout the study.

Any other information on results incl. tables

Table 1: Body weight (kg)

		Observation period
Animal #	Day of treatment	24h	48h	72h	7d	14d
1997	3.49	3.52	3.54	3.52	3.65	3.76
2011	3.21	3.23	3.29	3.27	3.39	3.51
2012	3.06	3.04	3.09	3.08	3.18	3.29

Table 2: Primary Eye Irritation Test

Animal #	Region of eye		Observation period
			1h	24h	48h	72h	7d	14d
1997	Cornea	Density	0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	Conjunctivae	Hyperaemia	1	1	1	0	0	0
		Edema	1	1	0	0	0	0
2011	Cornea	Density	0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	Conjunctivae	Hyperaemia	2	2	1	0	0	0
		Edema	2	1	0	0	0	0
2012	Cornea	Density	0	0	0	0	0	0
	Iris		0	0	0	0	0	0
	Conjunctivae	Hyperaemia	2	2	2	1	1	0
		Edema	3	2	1	0	0	0

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In an acute eye irritation/corrosion study conducted according to OECD 405, the test item is considered to be non-irritant.

Executive summary:

In a primary eye irritation study (OECD 405) 0.1 mL of the test item was instilled into the conjunctival sac of one eye of each of three young adult New Zealand White rabbits. Animals were observed for 14 days. Irritation was scored by the method of Draize. The behaviour and physical state of the rabbits, and the body-weight evolution were normal throughout the study. Only very mild conjunctival reactions were observed, which were fully reversible within 72 hours (two animals), and for the third animal within 14 days. Based on the results, the test item can be considered as not irritating.