Registration Dossier

Administrative data

Endpoint:
skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
08 August 2017 to 20 October 2017
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
guideline study

Data source

Reference
Reference Type:
study report
Title:
Unnamed
Year:
2017
Report date:
2018

Materials and methods

Test guidelineopen allclose all
Qualifier:
according to guideline
Guideline:
OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)
Version / remarks:
2015
Deviations:
no
Qualifier:
according to guideline
Guideline:
EU Method B.46 (In Vitro Skin Irritation: Reconstructed Human Epidermis Model Test)
Version / remarks:
2009
Deviations:
no
GLP compliance:
yes (incl. QA statement)
Remarks:
OGYI/38593-5/2012

Test material

Constituent 1
Chemical structure
Reference substance name:
2,20:3,19-Dimethano-2,3,4a,5a,6a,7a,8a,9a,10a,11a, 12a,13a,14a,15a,16a,17a,19 ,20,21a,22a, 23a,24a,25a,26a,27a,28a,29a,30a,31a,32a,33a,34a-dotriacontaazabispentaleno [1''',6''':5'',6'',7''] cycloocta[1'',2'',3'':3'',4'']pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno [1',6':5,6,7]cycloocta[1,2,3-gh:1',2',3'-g'h']cycloocta[1,2,3-cd:5,6,7-c'd']dipentalene-1,4,6,8,10,12, 14,16,18,21,23,25,27,29,31,33-hexadecone, hexadecahydro-, stereoisomer,2,18:3,17-Dimethano-2,3, ,4a,5a,6a,7a,8a,9a,10a,11a,12a,13a,14a,15a,17,18,19a, 20a,21a,22a,23a,24a, 25a,26a,27a,28a,29a,30a-octacosaazabispentaleno[1''',6''':5'',6'',7'']cycloocta [1'',2'',3'':3'',4''] pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene- 1,4,6,8,10,12,14,16,19,21,23,25,27,29-tetradecone, tetradecahydro-, stereoisomer, 1H,4H,12H,15H-2,14:3,13-Dimethano-5H,6H,7H,8H,9H,10H,11H,16H,17H,18H,19H,20H,21H,22H-2,3,4a, 5a,6a,7a,8a,9a,10a,11a,13,14,15a,16a,17a,18a,19a,20a,21a,22a-eicosaazabispentaleno [1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene- 1,4,6,8,10,12,15,17,19,21-decone, decahydro-, stereoisomer
EC Number:
946-188-6
Molecular formula:
(C6H6N4O2)n, n=6,7,8
IUPAC Name:
2,20:3,19-Dimethano-2,3,4a,5a,6a,7a,8a,9a,10a,11a, 12a,13a,14a,15a,16a,17a,19 ,20,21a,22a, 23a,24a,25a,26a,27a,28a,29a,30a,31a,32a,33a,34a-dotriacontaazabispentaleno [1''',6''':5'',6'',7''] cycloocta[1'',2'',3'':3'',4'']pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno [1',6':5,6,7]cycloocta[1,2,3-gh:1',2',3'-g'h']cycloocta[1,2,3-cd:5,6,7-c'd']dipentalene-1,4,6,8,10,12, 14,16,18,21,23,25,27,29,31,33-hexadecone, hexadecahydro-, stereoisomer,2,18:3,17-Dimethano-2,3, ,4a,5a,6a,7a,8a,9a,10a,11a,12a,13a,14a,15a,17,18,19a, 20a,21a,22a,23a,24a, 25a,26a,27a,28a,29a,30a-octacosaazabispentaleno[1''',6''':5'',6'',7'']cycloocta [1'',2'',3'':3'',4''] pentaleno[1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene- 1,4,6,8,10,12,14,16,19,21,23,25,27,29-tetradecone, tetradecahydro-, stereoisomer, 1H,4H,12H,15H-2,14:3,13-Dimethano-5H,6H,7H,8H,9H,10H,11H,16H,17H,18H,19H,20H,21H,22H-2,3,4a, 5a,6a,7a,8a,9a,10a,11a,13,14,15a,16a,17a,18a,19a,20a,21a,22a-eicosaazabispentaleno [1'',6'':5',6',7']cycloocta[1',2',3':3',4']pentaleno[1',6':5,6,7]cycloocta[1,2,3-cd:1',2',3'-gh]pentalene- 1,4,6,8,10,12,15,17,19,21-decone, decahydro-, stereoisomer
Test material form:
solid
Details on test material:
purity >85 % (w/w)

In vitro test system

Test system:
human skin model
Source species:
human
Cell type:
non-transformed keratinocytes
Vehicle:
other: test item was wetted with Dulbecco's Phosphate Buffered Saline
Control samples:
yes, concurrent negative control
yes, concurrent positive control
Amount/concentration applied:
Approximately 25 mg (~ 39 mg/cm2) of the test item were applied to the tissues, wetted with 25 µL DPBS (Dulbecco's Phosphate Buffered Saline) prior to application, and spread to match the surface of the tissue
Duration of treatment / exposure:
60 minutes
Number of replicates:
Three

Results and discussion

In vitro

Results
Irritation / corrosion parameter:
% tissue viability
Run / experiment:
1
Value:
84.4
Negative controls validity:
valid
Positive controls validity:
valid
Other effects / acceptance of results:
OTHER EFFECTS:
- Visible damage on test system: No
- Direct-MTT reduction: No
- Colour interference with MTT: No

DEMONSTRATION OF TECHNICAL PROFICIENCY: Yes

ACCEPTANCE OF RESULTS:
- Acceptance criteria met for negative control: yes
- Acceptance criteria met for positive control: yes (treatment with the positive control induced a decrease in the relative absorbance compared with the negative control to 4.2% thus ensuring the validity of the test system)
- Acceptance criteria met for variability between replicate measurements: yes
- Range of historical values if different from the ones specified in the test guideline: historical values were within the range specified in the test guideline

Any other information on results incl. tables

Results after treatment with Reaction mass of Cucurbit[6]uril, Cucurbit[7]uril and Cucurbit[8]uril and the controls

Dose Group

Tissue No.

Absor-bance 570 nm
Well 1

Absor-bance 570 nm
Well 2

Absor-bance 570 nm
Well 3

Mean Absor-bance of 3 Wells

Mean Absor-bance

of three wells blank

corrected

Mean

Absor-bance

of 3 tissues

after blank correction

Rel. Absor-bance [%] Tissue 1, 2 + 3*

Standard Deviation

[%]

Mean Rel. Absorbance (Viability)

[%]

Blank

 

0.039

0.038

0.044

0.040

0.000

 

 

 

 

Negative Control

1

1.396

1.399

1.398

1.398

1.357

1.495

90.8

8.1

100.0

2

1.580

1.593

1.581

1.584

1.544

103.3

3

1.631

1.626

1.612

1.623

1.583

105.9

Positive Control

1

0.103

0.105

0.104

0.104

0.064

0.063

4.3

3.6

4.2

2

0.104

0.105

0.105

0.104

0.064

4.3

3

0.100

0.101

0.100

0.100

0.060

4.0

Test Item

1

1.273

1.302

1.274

1.283

1.243

1.262

83.2

6.2

84.4

2

1.241

1.214

1.249

1.235

1.194

79.9

3

1.406

1.402

1.355

1.388

1.347

90.1

Applicant's summary and conclusion

Interpretation of results:
GHS criteria not met
Conclusions:
The mean relative absorbance value of the test item, corresponding to the cell viability, decreased to 84.4%. The test item therefore was "non-irritant" under the conditions of this in vitro test.
Executive summary:

The purpose of this study was to classify the skin irritation potential of the test item "reaction mass of cucurbit(6]uril, cucurbit[7]uril and cucurbit[8]uril". The study was performed in accordance with OECD test guideline 439 of July 2015 ("In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method”). Following exposure to the test item, the mean cell viability was 84.4% compared to the negative control. This is above the threshold of 50%, therefore the test item was considered as being non-irritant to skin. The experiment met the validity criteria, therefore the study was considered to be valid.