Amoxicillin

Endpoint summary

Description of key information

Skin irritation/corrosion: Test method OECD 439. GLP study. The substance was determined to be not irritating to the skin.
Eye damage: Test method OECD 438. GLP study. The substance did not cause eye damage. Nevertheless, no categorization was possible.
Eye irritation/corrosion: Test method OECD 405. The substance was determined to be not irritating to the eye.

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method according to OECD 439. GLP study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

other: in-vitro

Strain:

other: in-vitro

Type of coverage:

open

Preparation of test site:

other: Not required.

Vehicle:

water

Controls:

yes

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied (volume or weight with unit): 20 mg
- Concentration (if solution): As the test item was solid, first 10 µL distilled water was applied to the epidermal surface and then 20 mg of the test item.

VEHICLE
- Amount(s) applied (volume or weight with unit): 10µL.
- Concentration (if solution): 20mg/10 µL.

Duration of treatment / exposure:

15 minuts.

Number of animals:

Not applicable.

Details on study design:

TEST SITE
- Area of exposure: 0.38cm2.

REMOVAL OF TEST SUBSTANCE
- Washing (if done): PBS
- Time after start of exposure: 15 min.

Irritant / corrosive response data:

Amoxicillin trihydrate, the results indicate that the test item is non-irritant to skin (No Category).

Substance	Optical density (OD)			Viability (% RV)
		Measured	Blank corrected
Negative control Phosphate buffered saline	1	0.934	0.887	95.8
	2	0.971	0.924	99.7
	3	1.015	0.968	104.5
	Mean	--	0.927	100.0
Positive control 5% (w/v) SDS solution	1	0.108	0.061	6.6
	2	0.096	0.049	5.3
	3	0.083	0.036	3.9
	Mean	--	0.049	5.3
Amoxicillin trihydrate	1	0.847	0.800	86.3
	2	0.91	0.864	93.2
	3	0.884	0.837	90.4
	Mean	--	0.934	90.0

Interpretation of results:

not classified

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Following an exposure to amoxicillin trihydrate the mean cell viability of the skin disks was 90% compared to the negative control. The test item was considered to be non-irritant to the skin (No category).

Executive summary:

A in vitro skin irritation test was conducted according to the OECD guideline 439 under GLP conditions. 3 Disks of EpiSkin-SM were treated with the test subtance amoxicillin trihydrate and incubated for 15 minutes at room temperature. Exposure was terminated by rinsing with PBS, the epidermis units were then incubated at 37ºC for 42 hours. The viability of each disk was assessed through the MTT assay: tissues were incubated for 3 hours with a MTT solution at 37ºC protected from light. The precipitated formazan crystals were extracted using acidified isopropanol and were quantified spectrophotometrically. PBS and 5 % Sodium Dodecyl Sulphate (SDS) were used as negative and positive control respectively. An additional disk was used to provide and estimate of color contribution from the test item. The viability was expressed as a % relative to the negative control. Following exposure to amoxicillin trihydrate the mean cell viability was 90%, this is above the threshold of 50% adn according to OECD 439 guideline the test was considered to be non-irritant to the skin (No category). The study meet the validity criteria.

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

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Test method according to OECD 405. GLP Study.

Qualifier:

according to guideline

Guideline:

OECD Guideline 405 (Acute Eye Irritation / Corrosion)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method B.5 (Acute Toxicity: Eye Irritation / Corrosion)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Species:

rabbit

Strain:

New Zealand White

Details on test animals or tissues and environmental conditions:

TEST ANIMALS
- Source: S&K-LAP Kft., 2173 Kartal, Császár út 135, Hungary
- Age at study initiation: 13 - 14 weeks.
- Weight at study initiation: 3312 - 3840 g.
- Housing: Individually housed in AAALAC approved metal wire rabbit cages. Cages were of an open wire structure and cages were placed together to allow some social interaction with rabbits in adjoining cages.
- Diet: UNI diet for rabbits ad libitum
- Water: Tap water as for human consumption ad libitum
- Acclimation period: At least 19 days.

ENVIRONMENTAL CONDITIONS
- Temperature (°C): 17.4 - 22.1 ºC
- Humidity (%): 24 - 48 %
- Air changes (per hr): 15-20 air exchanges/hour
- Photoperiod: 12 hours light/12 hours dark

Vehicle:

unchanged (no vehicle)

Controls:

other: The untreated contralateral eye served as the control.

Amount / concentration applied:

TEST MATERIAL
- Amount(s) applied: 0.1 g

Duration of treatment / exposure:

As the solid test item remained in the eye sac at the one hour observation time point, the treated eye of test animals was rinsed with physiological saline solution in all animals.

Observation period (in vivo):

3 days, the duration of the observation period was sufficient to identify reversibility of changes.

Number of animals or in vitro replicates:

3

Details on study design:

REMOVAL OF TEST SUBSTANCE
- Washing (if done): Physiological saline solution.
- Time after start of exposure: 1 hour.

SCORING SYSTEM: According to Draize system and OECD 405 (2012).

TOOL USED TO ASSESS SCORE: expert evaluation, nude eye.

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #1

Time point:

other: 1 hour

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: 24, 48 and 72 hours after the application the score was 0

Irritation parameter:

chemosis score

Basis:

animal #1

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #1

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #1

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal #1

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #2

Time point:

other: 1 hour

Score:

0

Max. score:

0

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: 24, 48 and 72 hours after the application the score was 0

Irritation parameter:

chemosis score

Remarks:

Chemosis

Basis:

animal #1

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #2

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #2

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal #2

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Redness

Basis:

animal #3

Time point:

other: 1

Score:

1

Max. score:

1

Reversibility:

fully reversible within: 24 hours

Remarks on result:

other: 24, 48 and 72 hours after the application the score was 0

Irritation parameter:

chemosis score

Basis:

animal #3

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

conjunctivae score

Remarks:

Discharge

Basis:

animal #3

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

cornea opacity score

Basis:

animal #3

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritation parameter:

iris score

Basis:

animal #3

Time point:

other: 1, 24, 48 and 72 hours

Score:

0

Max. score:

0

Irritant / corrosive response data:

The only finding was conjunctival redness (score 1= some blood vessels hyperaemic) in the three animals one hour after the application of the test item. The effect was fully reversible within 24 hours.

Other effects:

The test item remained in the conjunctival sac one hour after its application.

SCORING AND ASSESSMENT OF LOCAL REACTION 

1. Conjunctivae

 

A. Redness (Palpebral and bulbar)

Normal: 0

Some blood vessels hyperaemic (injected): 1

Diffuse, crimson colour, individual vessels not easily discernible: 2

Diffuse beefy red: 3

 

B. Chemosis

Normal: 0

Some swelling above normal (includes nictating membrane): 1

Obvious swelling with partial eversion of lids: 2

Swelling with lids about half closed: 3

Swelling with lids more than half closed: 4

 

C. Discharge

No discharge: 0

Any amount different from normal (does not include small amounts observed in inner canthus of normal animals): 1

Discharge with moistening of the lids and hairs just adjacent to lids: 2

Discharge with moistening of the lids and hairs, on considerable area

around the eye: 3

 

2. Iris

 

D. Values

Normal: 0

Markedly deepened rugae, congestion, swelling, moderate circumcorneal hyperaemia: or injection: iris reactive to light (a sluggish reaction is considered to be an effect): 1

Haemorrhage, gross destruction, or no reaction to light: 2

 

3. Cornea

 

E. Opacity-degree of density (Area most dense taken for reading)

No ulceration or opacity: 0

Scattered or diffuse areas of opacity (other than slight dulling of normal lustre): details of iris clearly visible: 1                              

Easily discernible translucent area: details of iris slightly obscured: 2

Nacrous area: no details of iris visible: size of pupil barely discernible: 3

Opaque cornea: iris not discernible through the opacity: 4

 

F. Area of cornea involved

One quarter (or less), but not zero: 1

Greater than one quarter, but less than half : 2

Greater than half, but less than three quarters: 3

Greater than three quarters, up to whole area: 4

 

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Amoxicillin trihdyrate is not irritating. According to the test results, amoxicillin is not irritating.

Executive summary:

An acute eye irritation study of the test item Amoxicillin trihydrate was performed in three New Zealand White rabbits. The irritation effects of the test item were evaluated according to the Draize method (OECD No.: 405, 2012). Rabbits were treated with analgesic and anaesthetic as per the regulatory guideline. Three animals were used to make the classification. The test item was placed into the conjunctival sac of the left eye of each animal. The untreated right eye served as control. A single amount of 0.1 g of the test item was administered as a single dose. The eyes were examined at 1, 24, 48 and 72 hours after application. The test item Amoxicillin trihydrate, applied to rabbit eye mucosa, caused minor transient conjunctival effects at one hour after application which were fully reversible within 24 hours. According to Regulation (EC) No 1272/2008, Amoxicillin trihydrate does not require classification as an eye irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Additional information

Skin irritation/corrosion

Key study: A in vitro skin irritation test was conducted according to the OECD guideline 439 under GLP conditions. Following exposure to amoxicillin trihydrate the mean cell viability was 90%, this is above the threshold of 50% adn according to OECD 439 guideline the test was considered to be non-irritant to the skin (No category). The study meet the validity criteria.

Eye irritation/corrosion:

Key study: The study of eye irritation/corrosion of the test substance was performed in vitro according to OECD 438, with GLP. Amoxicillin trihydrate did not cause eye damage. According to UN GHS classification criteria, No prediction can be made, since the ICE Class combination of the 3 endpoints were 1 x I, 2 x II (the first run and the second run).

Key study: An acute eye irritation study of the test item Amoxicillin trihydrate was performed in three New Zealand White rabbits. The irritation effects of the test item were evaluated according to the Draize method (OECD 405, GLP). The test item applied to rabbit eye mucosa, caused minor transient conjunctival effects at one hour after application which were fully reversible within 24 hours. According to Regulation (EC) No 1272/2008, Amoxicillin trihydrate does not require classification as an eye irritant.

Justification for selection of skin irritation / corrosion endpoint:
Only one study available.

Justification for selection of eye irritation endpoint:
The in-vivo study is selected.

Justification for classification or non-classification

Based on available information, the substance is not classified for skin and eye irritation/corrosion according to Regulation (EC) no. 1272/2008.