1,3-dichloropropan-2-ol

Endpoint summary

Description of key information

The tissue viabilities observed in an in vitro skin corrosion study after 3-minute and 1-hour treatments with the test item

compared to the negative control tissues was 77% and 6.4%, respectively.  Because the mean relative tissue viability for the test item was below 15% after the 1-hour treatment it is considered to be corrosive and fulfils the classification criteria for Skin corrosion Category 1in accordance with the the criteria set out in 1272/2008/EC.

In the Bovine Corneal Opacity and Permeability assay, the substance induced an IVIS ≥ 55, it is concluded that the substance induces serious eye damage and should be classified category 1 according to the criteria set out in 1272/2008/EC.

Based upon the corrosive properties of the substance to both skin and mucosal membranes it is anticipated that the substance wil be a respiratory irritant and is classified as STOT SE H335. The substance is reported to be irritating to mucosal membranes (Lewis RJ, 2005)

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 431 (In Vitro Skin Corrosion: Reconstructed Human Epidermis (RHE) Test Method)

Deviations:

yes

Remarks:

coefficient of variation of viability of test item treated tissues @3 minutes = 31% = above acceptance criteria- did not affect outcome

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: 1,3-dichloro-2-propanol
Appearance: Colourless liquid
Batch: ZMG-197685
Purity/Composition: 99.2%
Test item storage: At room temperature container flushed with
nitrogen
Stable under storage conditions until: 09 December 2017 (expiry date)

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from a single donor

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm Skin Model (EPI-200, Lot no.: 26767, Appendix 4).
The model consists of normal, human-derived epidermal keratinocytes which have been
cultured to form a multilayered, highly differentiated model of the human epidermis. It
consists of organized basal, spinous and granular layers, and a multi-layered stratum corneum
containing intercellular lamellar lipid layers arranged in patterns analogous to those found in
vivo. The EpiDerm tissues (surface 0.6 cm²) were cultured on polycarbonate membranes of
10 mm cell culture inserts.

Control samples:

yes, concurrent negative control

yes, concurrent positive control

yes, concurrent MTT non-specific colour control

Amount/concentration applied:

50µl per well

Duration of treatment / exposure:

3 minutes and 1 hour

Duration of post-treatment incubation (if applicable):

3 hours for MTT metabolism

Number of replicates:

two tissues per timepoint

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minutes

Value:

ca. 77

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hour

Value:

ca. 6.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The test item was checked for color interference in aqueous conditions and possible direct
MTT reduction by adding the test item to MTT medium. Because the solutions did not turn
blue / purple nor a blue / purple precipitate was observed it was concluded that the test item
did not interfere with the MTT endpoint.

The relative mean tissue viability obtained after the 3-minute and 1-hour treatments with the test item compared to the negative
control tissues was 77% and 6.4% respectively. Because the mean relative tissue viability for
the test item was below 15% after 1 hour treatment it is considered to be corrosive.
The absolute mean OD570 (optical density at 570 nm) of the negative control tissues was
within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance
limit <=2.8) and the laboratory historical control data range. The mean
relative tissue viability following the 1-hour exposure to the positive control was 9.2%.
In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was
<= 16% for the negative control, indicating that the test system functioned properly

Interpretation of results:

Category 1 (corrosive) based on GHS criteria

Conclusions:

In conclusion, 1,3-dichloro-2-propanol is corrosive in the in vitro skin corrosion test under the
experimental conditions described in this report.

Executive summary:

The objective of this study was to evaluate 1,3-dichloro-2-propanol for its ability to induce

skin corrosion on a human three dimensional epidermal model (EpiDerm (EPI-200)).  The

possible corrosive potential of the test item was tested through topical application for

3 minutes and 1 hour.

The study procedures described in this report were based on the most recent OECD and EC

guidelines.

Batch ZMG-197685 of the test item was a colourless liquid. The test item was applied

undiluted (50 µl) was applied directly on top of the skin tissue.  

The positive control had a mean relative tissue viability of 9.2% after the 1-hour exposure.

The absolute mean OD570

(optical density at 570 nm) of the negative control tissues was

within the acceptance limits of OECD 431 (lower acceptance limit ≥0.8 and upper acceptance

limit 2.8) and the laboratory historical control data range.  In the range of 20 - 100%

viability the Coefficient of Variation between tissue replicates was  16% for the negative

control, indicating that the test system functioned properly.

Skin corrosion is expressed as the remaining cell viability after exposure to the test item.  The

relative mean tissue viability obtained after 3-minute and 1-hour treatments with the test item

compared to the negative control tissues was 77% and 6.4%, respectively.  Because the mean

relative tissue viability for the test item was below 15% after the 1-hour treatment it is

considered to be corrosive.  

In conclusion, 1,3-dichloro-2-propanol is corrosive in the in vitro skin corrosion test under the

experimental conditions described in this report.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (corrosive)

Eye irritation

Link to relevant study records

Reference

Endpoint:

eye 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:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 437 (Bovine Corneal Opacity and Permeability Test Method for Identifying i) Chemicals Inducing Serious Eye Damage and ii) Chemicals Not Requiring Classification for Eye Irritation or Serious Eye Damage)

Qualifier:

according to guideline

Guideline:

EU method B.47 (Bovine corneal opacity and permeability test method for identifying ocular corrosives and severe irritants)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Identification: 1,3-dichloro-2-propanol
Appearance: Colourless liquid
Batch: ZMG-197685
Purity/Composition: 99.2%
Test item storage: At room temperature container flushed with
nitrogen
Stable under storage conditions until: 09 December 2017 (expiry date)

Species:

cattle

Details on test animals or tissues and environmental conditions:

Test System: Bovine eyes were used as soon as possible after slaughter.
Rationale: In the interest of sound science and animal welfare, a sequential
testing strategy is recommended to minimize the need of in vivo
testing. As a consequence a validated and accepted in
vitro test for eye irritation should be performed before in vivo
tests are conducted. One of the proposed validated in vitro eye
irritation tests is the Bovine Corneal Opacity and Permeability
(BCOP) test.
Source: Bovine eyes from young cattle were obtained from the
slaughterhouse (Vitelco, 's Hertogenbosch, The Netherlands),
where the eyes were excised by a slaughterhouse employee as
soon as possible after slaughter.
Transport: Eyes were collected and transported in physiological saline in a
suitable container under cooled conditions.

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

750µL per cornea

Duration of treatment / exposure:

10 minutes

Duration of post- treatment incubation (in vitro):

120 minutes +- 10 minutes

Number of animals or in vitro replicates:

3 (three)

Details on study design:

SELECTION AND PREPARATION OF CORNEAS
he eyes were checked for unacceptable defects, such as opacity, scratches, pigmentation and neovascularization by removing them from the physiological saline and holding them in the light. Those exhibiting defects were discarded.
The isolated corneas were stored in a petri dish with cMEM (Earle’s Minimum Essential
Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine
(Life Technologies) and 1% (v/v) Foetal Bovine Serum (Life Technologies)). The isolated
corneas were mounted in a corneal holder (one cornea per holder) of BASF (Ludwigshafen, Germany) with the endothelial side against the O-ring of the posterior half of the holder.
The anterior half of the holder was positioned on top of the cornea and tightened with screws.
The compartments of the corneal holder were filled with cMEM of 32 +- 1C. The corneas
were incubated for the minimum of 1 hour at 32+- 1C.

QUALITY CHECK OF THE ISOLATED CORNEAS

After the incubation period, the medium was removed from both compartments and replaced with fresh cMEM. Opacity determinations were performed on each of the corneas using an opacitometer (BASF-OP3.0, BASF, Ludwigshafen, Germany). The opacity of each cornea was read against a cMEM filled chamber, and the initial opacity reading thus determined was recorded. Corneas that had an initial opacity reading higher than 7 were not used.

NUMBER OF REPLICATES

Three corneas were selected at random for each treatment group.

NEGATIVE CONTROL USED
cMEM (Earle’s Minimum Essential
Medium (Life Technologies, Bleiswijk, The Netherlands) containing 1% (v/v) L-glutamine
(Life Technologies) and 1% (v/v) Foetal Bovine Serum (Life Technologies))
SOLVENT CONTROL USED (if applicable)


POSITIVE CONTROL USED
96% v/v Ethanol

APPLICATION DOSE AND EXPOSURE TIME
750 µl, 10 minutes
TREATMENT METHOD: [closed chamber / open chamber]
closed chamber

POST-INCUBATION PERIOD: no.

REMOVAL OF TEST SUBSTANCE
- two- MEM (earles alts with phenol red, then cMEM

- POST-EXPOSURE INCUBATION:
120 minutes +- 10 minutes ( opacity); 90 +- 5 minutes fluorescein

METHODS FOR MEASURED ENDPOINTS:
- Corneal opacity: YES
- Corneal permeability: passage of sodium fluorescein dye measured with the aid of [UV/VIS spectrophotometry / microtiter plate reader] (OD490) YES
- Others (e.g, pertinent visual observations,

SCORING SYSTEM: In Vitro Irritancy Score (IVIS)

DECISION CRITERIA: decision criteria specified in the TG used.

Irritation parameter:

cornea opacity score

Run / experiment:

cornea 1

Value:

39.7

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

cornea opacity score

Run / experiment:

cornea 2

Value:

41.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

cornea opacity score

Run / experiment:

cornea 3

Value:

44.6

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein leakage

Run / experiment:

cornea 1

Value:

2.658

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein leakage

Run / experiment:

cornea 2

Value:

1.712

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

fluorescein leakage

Run / experiment:

cornea 3

Value:

2.758

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea 1

Value:

80

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea 2

Value:

67

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Irritation parameter:

in vitro irritation score

Run / experiment:

cornea 3

Value:

86

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

The negative control responses for opacity and permeability were less than the upper limits of
the laboratory historical range indicating that the negative control did not induce irritancy on
the corneas. The mean in vitro irritancy score of the positive control (Ethanol) was 61 and
within two standard deviations of the current historical positive control mean.
It was therefore concluded that the test conditions were adequate and that the test system
functioned properly.
1,3-dichloro-2-propanol induced serious eye damage through both endpoints, resulting in a
mean in vitro irritancy score of 78 after 10 minutes of treatment.

Interpretation of results:

Category 1 (irreversible effects on the eye) based on GHS criteria

Conclusions:

In conclusion, since 1,3-dichloro-2-propanol induced an IVIS ≥ 55, it is concluded that
1,3-dichloro-2-propanol induces serious eye damage in the Bovine Corneal Opacity and
Permeability test under the experimental conditions described in this report and should be
classified category 1 according to the Globally Harmonized System of Classification and
Labeling of Chemicals (GHS) of the United Nations.

Executive summary:

The objective of this study was to evaluate the eye hazard potential of  

1,3-dichloro-2-propanol as measured by its ability to induce opacity and increase permeability

in an isolated bovine cornea using the Bovine Corneal Opacity and Permeability test (BCOP

test).

This report describes the potency of chemicals to induce serious eye damage using isolated

bovine corneas.  The eye damage of the test item was tested through topical application for

10 minutes.  

The study procedures described in this report were based on the most recent OECD guideline.

Batch ZMG-197685 of the test item was a colourless liquid.  The test item was applied as it is

(750 µl) directly on top of the corneas.

The negative control responses for opacity and permeability were less than the upper limits of

the laboratory historical range indicating that the negative control did not induce irritancy on

the corneas.  The mean in vitro irritancy score of the positive control (Ethanol) was 61 and

was within two standard deviations of the current historical positive control mean.  It was

therefore concluded that the test conditions were adequate and that the test system functioned

properly.  

1,3-dichloro-2-propanol induced serious eye damage through both endpoints, resulting in a

mean in vitro irritancy score of 78 after 10 minutes of treatment.

In conclusion, since 1,3-dichloro-2-propanol induced an IVIS ≥ 55, it is concluded that 1,3dichloro-2-propanol

induces serious eye damage in the Bovine Corneal Opacity and Permeability test under the experimental conditions described

in this report and should be classified category 1 according to the Globally Harmonized System of Classification and Labeling of Chemicals

(GHS) of the United Nations.

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irreversible damage)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

adverse effect observed (irritating)

Justification for classification or non-classification

The tissue viabilities observed in an in vitro skin corrosion study after 3-minute and 1-hour treatments with the test item

compared to the negative control tissues was 77% and 6.4%, respectively.  Because the mean relative tissue viability for the test item was below 15% after the 1-hour treatment it is considered to be corrosive and fulfils the classification criteria for Skin corrosion Category 1 in accordance with the

the criteria set out in 1272/2008/EC.

In the Bovine Corneal Opacity and Permeability assay, the substance induced an IVIS ≥ 55, it is concluded that the substance induces serious eye damage and should be classified category 1 according to the criteria set out in 1272/2008/EC.Following reports that the substances is irritating to mucosal memebranes (Lewis RJ, 2005) and based upon the corrosive properties of the substance to both skin and mucosal membranes it is anticipated that the substance wil be a respiratory irritant and is classified as STOT SE H335.