Amoxicillin

Skin irritation / corrosion

Administrative data

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.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2016

Report date:

2016

Materials and methods

Test guideline

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)

Test material

Test material information

Constituent 1

Reference substance name:

(2S,5R,6R)-6-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid trihydrate.

Cas Number:

61336-70-7

Molecular formula:

C16H25N3O8S

IUPAC Name:

(2S,5R,6R)-6-[[(2R)-2-amino-2-(4-hydroxyphenyl)acetyl]amino]-3,3-dimethyl-7-oxo-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid trihydrate.

Constituent 2

Reference substance name:

Amoxicillin trihydrate

IUPAC Name:

Amoxicillin trihydrate

Test material form:

solid: particulate/powder

Remarks:

migrated information: powder

Details on test material:

- Name of test material: Amoxicillin trihydrate
- Physical state: White powder.
- Analytical purity: 98.7%
- Purity test date: 13 October 2015
- Lot/batch No.: B331003
- Expiration date of the lot/batch: September 2020
- Storage condition of test material: Controlled room temperature (15-25ºC, below 70 RH %)

Test animals

Species:

other: in-vitro

Strain:

other: in-vitro

Test system

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.

Results and discussion

In vivo

Irritant / corrosive response data:

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

Any other information on results incl. tables

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

Applicant's summary and conclusion

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.