N-acetyl-β-D-glucosamine

Skin irritation / corrosion

Administrative data

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

25th July 2019 - 23rd December 2019

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2019

Report date:

2019

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:

N-acetyl-β-D-glucosamine

EC Number:

231-368-2

EC Name:

N-acetyl-β-D-glucosamine

Cas Number:

7512-17-6

Molecular formula:

C8H15NO6

IUPAC Name:

N-acetyl-β-D-glucosamine

Test material form:

solid

Specific details on test material used for the study:

Analysed concentration 99.42%
Manufactured by R. N. Laboratories Pvt. Ltd.
Appearance: White powder
Date of manufacture: May 2019, Date of expiry: April 2022
Batch No.: RD/NAG/19/E-006
pH 6.75 (limit: 6.0 - 8.0)
Stored at 4°C in a light resistance container

In vitro test system

Test system:

artificial membrane barrier model

Source species:

other: Reconstructed human epidermis (SkinEthic™ RHE).

Cell type:

other: normal human keratinocytes

Vehicle:

unchanged (no vehicle)

Details on test system:

Reconstructed human epidermis (SkinEthic(TM) RHE). Normal human keratinocytes cultured for 17-days on a 0.5 cm^2 polycarbonate filter insert at the air-liquid interface in a chemically defined growth medium. The cells form a multi-layered, highly differentiated and stratified epidermis model of the human epidermis that consists of a main basal, supra basal, spinous and granular layers and a functional stratum corneum

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

TEST MATERIAL
- NADG
-16 ± 2 mg of test item/0.5 cm2

NEGATIVE CONTROL
- Dulbecco’s Phosphate Buffered Saline (DPBS)
- 16 µL/0.5 cm2

POSITIVE CONTROL
- 5% Sodium dodecyl sulfate, 5% aqueous
- 16 µL/0.5 cm2

Duration of treatment / exposure:

42 minutes

Duration of post-treatment incubation (if applicable):

Tissues were incubated in 6-well plates containing 2mL growth medium at 37 ± 1°C in 5 ± 1% CO2 in a humidified incubator for 42 hours.

Number of replicates:

3

Results and discussion

In vitro

Results

Irritation / corrosion parameter:

% tissue viability

Remarks:

mean percent viability

Value:

95.5

Positive controls validity:

valid

Remarks:

A reduction in the cell viability of 1.2 %

Remarks on result:

no indication of irritation

Any other information on results incl. tables

There was a significant difference observed in the absorbance for colour interference between Isopropanol (negative control) and the test item:

Treatment	Optical Density (nm)	Interaction
Negative Control (Isopropanol)	0.042	No
	0.048
N-Acetyl-D-Glucosamine	0.049	No
	0.061

NADG did not cause direct MTT reduction compared to the concurrent negative control (maintainence medium):

Treatment	Interaction
Negative Control (Maintenance Medium)	No
N-Acetyl-D-Glucosamine	No

N-Acetyl-D-Glucosamine is classified as "No Category (Non Skin Irritant)" according to the results of this study.

Data Summary of Percent Viability

Treatment	Tissue Replicate	OD at 570 nm	Blank Corrected OD	Mean of Corrected OD	Mean OD of Three Tissues	% Viability/ Tissue	Mean % Viability	SD of % Viability	CV of % Viability	Corrosivity Class
Negative Control (Dulbecco’s Phosphate Buffered Saline (DPBS))	1	1.679	1.636	1.548	1.644	100	100	0.084	5.11	NA
		1.737	1.694
		1.356	1.313
	2	1.876	1.833	1.703
		1.728	1.685
		1.633	1.590
	3	1.722	1.679	1.681
		1.720	1.677
		1.730	1.687
N-Acetyl-D-Glucosamine	1	1.680	1.633	1.571	1.570	95.6	95.5	1.852	1.94	No Category
		1.596	1.549
		1.579	1.532
	2	1.596	1.549	1.538		93.6
		1.545	1.498
		1.614	1.567
	3	1.654	1.607	1.600		97.3
		1.646	1.599
		1.642	1.595
Positive control (Sodium dodecyl sulphate (5% aq.))	1	0.062	0.019	0.02	0.020	1.2	1.2	0.058	4.83	Category 2
		0.063	0.020
		0.063	0.020
	2	0.064	0.021	0.02		1.2
		0.063	0.020
		0.063	0.020
	3	0.062	0.019	0.021		1.3
		0.064	0.021
		0.065	0.022

Results of the negative control met the OECD guideline 439 acceptance range for the prediction model SkinEthic^TM RHE. The positive control also met the acceptance criteria of the OECD guideline 439.

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

Based on results of this study, the classification for N-Acetyl-D-Glucosamine is No Category (Non Skin Irritant).

Executive summary:

Tissues were exposed to the negative control (Dulbecco’s Phosphate Buffered Saline (DPBS)), positive control (sodium dodecyl sulfate, 5% aqueous (SDS)) and the test item, N-Acetyl-D-Glucosamine in triplicate for 42 minutes, at the room temperature.

The mean cell viability in tissues treated with the test item was 95.5% after, 42 minutes’ exposure. No significant reduction in the percent cell viability was observed in treated tissues, when compared with that of the concurrent negative control. All criteria for a valid study were met.

Based on results of this study, the classification for N-Acetyl-D-Glucosamine is No Category (Non Skin Irritant).