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Administrative data

skin irritation: in vitro / ex vivo
Type of information:
experimental study
Adequacy of study:
key study
Study period:
2010-03-08 to 2010-06-10
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP study conducted according to EU method B.46

Data source

Reference Type:
study report
Report date:

Materials and methods

Test guideline
according to guideline
other: EU Method B.46 (In Vitro Skin Irritation: Reconstructed human epidermis model test)
Principles of method if other than guideline:
In addition the following guidelines were followed:
•OECD Guideline for the Testing of Chemicals, Draft Proposal for a New Guideline, In Vitro Skin Irritation: Reconstructed Human Epidermis (RhE) Test Method, Version 7.6, 9 September 2009
•ECVAM international validation study on in vitro tests for acute skin irritation: Report on the validity of the EPISKIN and EpiDerm assays and on the Skin Integrity Func-tion Test (Altern Lab Anim. 2007 Dec; 35 (6): 559-601).
•Protocol for IN VITRO EpiDermTM SKIN IRRITATION TEST (EPI-200-SIT), Rev. 3/23/2009, MatTek Corporation, Ashland, MA 01721, USA
GLP compliance:
yes (incl. QA statement)

Test material

Constituent 1
Chemical structure
Reference substance name:
EC Number:
EC Name:
Cas Number:
Molecular formula:
Details on test material:
- Name of test material (as cited in study report): 3-Chloro-1,2 propane diol
- Physical state: Liquid
- Composition of test material, percentage of components: 3-chloro-1,2-propane diol, > 98%
- Analytical purity: 99,3% (GC)
- Lot/batch No.: 5225
- Expiration date of the lot/batch: 17/11/2010
- Storage condition of test material: Stored in a closed vessel at room temperature protected from light
- Stability: unknown
- Other: date of production: 17 Nov 2009

Test animals

other: Not applicable (in vitro study)
other: Not applicable (in vitro study)
Details on test animals or test system and environmental conditions:
Test system: Commercially available Epi-200-kit
Source: MatTek Corporation, Ashland, USA
Day of delivery: 19-05-2010
Batch no.: 13614

Test system

Type of coverage:
other: Not applicable (in vitro study)
Preparation of test site:
other: Not applicable (in vitro study)
unchanged (no vehicle)
Amount / concentration applied:
30 µL of test item
Duration of treatment / exposure:
60 mins.
Observation period:
42 hours
Number of animals:
3 tissues
Details on study design:
Pre-incubated tissues were placed into fresh 6-well-plates containing 0.9 mL assay medium per well. Each plate contained three tissues whereas one plate was used as negative control, one as postive control and one as treatment with the test item. Tissues were dosed in one minute intervals and after dosing the last tissue placed into the incubator for 35 min. One hour after the first application, the inserts were removed from the plates and rinsed immediately in one-minute intervals. Tissues where then dried and transferred into a new 6-well-plate. After a total incubation time of 42 hours a MTT-reagent was added and allowed to incubate for 3 hours. Finally the MTT was aspirated and replaced with PBS buffer, which was aspirated and replaced by 2 ml of isopropanol. The next day the produced fromazan was collected, mixed to achieve homogenisation and transferred to a 96-well plate to beread in a plate spectral photometer at 570 nm. Two replicates per tissue were measured.

Results and discussion

In vitro

Irritation / corrosion parameter:
other: other: % formazan production mean
Remarks on result:
Basis: other: formazan production. Time point: 42 hrs. Max. score: 100.0. Remarks: values are expressed in %. (migrated information)

Any other information on results incl. tables

The pre test revealed that 3 -chloro-1,2 propane diol did not reduce formazan directly, nor did it form a colour without the addition of MTT.

In a first measurement the variation within the replicates (RSD) was 33.3%, exceeding the allowed maximum of 18% as described in the guideline. However it was considered that this was due to an incomplete homogenisation, whereafter a second measurment was perofmed after thorough mixing of the contents in each insert. The RSD in the second measurement was 11.7%, fulfilling the acceptance criteria as set out in the guideline. The experiments was considered valid based on the latter, the fact that both experiments were clearly considered as irritant and that all values for negative and positive controls were within the range of historical data.

Applicant's summary and conclusion

Interpretation of results:
Migrated information Criteria used for interpretation of results: EU
3-chloro-1.2 propane diol is concluded to be irritating in the current study according to EU method B.46. A positive result in this test method is accepted for the classification and labelling of skin irritants under GHS. Therfore the substance should be classified as Skin irritant cat. 2, H315 (under CLP) and Xi R38 (under DSD).

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