DL-α-(aminomethyl)-p-hydroxybenzylic alcohol hydrochloride

Skin irritation / corrosion

Administrative data

Endpoint:

skin corrosion: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

26-06-2018 to 21-08-2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

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

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

1

Reference substance name:

DL-α-(aminomethyl)-p-hydroxybenzylic alcohol hydrochloride

EC Number:

212-216-4

EC Name:

DL-α-(aminomethyl)-p-hydroxybenzylic alcohol hydrochloride

Cas Number:

770-05-8

Molecular formula:

C8H11NO2.ClH

IUPAC Name:

4-(2-amino-1-hydroxyethyl)phenol hydrochloride

Test material form:

solid: particulate/powder

Specific details on test material used for the study:

Identification: Octopamine hydrochloride
Appearance: White to off white powder
Batch: D151-1710037
Purity/Composition: 99.8%
Test item storage: At room temperature protected from light
Stable under storage conditions until: 26 October 2019 (retest date)

In vitro test system

Test system:

human skin model

Source species:

human

Cell type:

non-transformed keratinocytes

Cell source:

foreskin from a single donor

Source strain:

not specified

Details on animal used as source of test system:

Mattek corp: Human neonatal foreskin lot number 28831 keratinocyte strain number 00267

Justification for test system used:

Recommended test system in international guidelines (OECD and EC).

Vehicle:

unchanged (no vehicle)

Details on test system:

EpiDerm Skin Model (EPI-200, Lot no.: 28831, kit O&P,)
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

Amount/concentration applied:

28.7 to 33.8 mg of the solid test item was added into the 6-well plates on top of the
skin tissues.

Duration of treatment / exposure:

3 minutes and 1 hour

Duration of post-treatment incubation (if applicable):

N/A

Number of replicates:

Two tissues were used for a 3-minute exposure to Octopamine
hydrochloride and two for a 1-hour exposure.

Results and discussion

In vitro

Results

Results 1

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

3 minutes

Value:

ca. 94

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Results 2

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

1 hour

Value:

ca. 60

Vehicle controls validity:

valid

Negative controls validity:

valid

Positive controls validity:

valid

Other effects / acceptance of results:

Octopamine hydrochloride 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 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 7.4%.
In the range of 20 - 100% viability the Coefficient of Variation between tissue replicates was
<=13%, indicating that the test system functioned properly.

Any other information on results incl. tables

The mean absorption at 570 nm measured after treatment with Octopamine hydrochloride and

controls are presented in Table 1.  

Table 2 shows the mean tissue viability obtained after 3-minute and 1-hour treatments with

Octopamine hydrochloride compared to the negative control tissues.  Skin corrosion is

expressed as the remaining cell viability after exposure to the test item.  The relative mean

tissue viability obtained after the 3-minute and 1-hour treatments with Octopamine

hydrochloride compared to the negative control tissues was 94% and 60% respectively.

Because the mean relative tissue viability for Octopamine hydrochloride was not below 50%

after 3 minutes treatment and not below 15% after 1 hour treatment Octopamine

hydrochloride is considered to be not corrosive.

Table 1

Mean Absorption in the in vitro Skin Corrosion Test with Octopamine hydrochloride

	3 minute application				1 hour application
	A(OD570)	B(OD570)	Mean OD570	SD	A(OD570)	B(OD570)	Mean OD570	SD
Negative Control	1.724	1.762	1.743	0.027	1.814	1.715	1.765	0.070
Test Item	1.745	1.518	1.632	0.160	1.095	1.021	1.058	0.052
Positive Control	0.145	0.103	0.124	0.030	0.120	0.139	0.130	0.014

Table 2: mean Tissue Viability in the in vitro Skin corrosion test with Octopamine hydrochloride

	3-minute application viability (percentage of control)	1-hour application viability (percentage of control)
Negative control	100	100
Octopamine hydrochloride	94	60
Positive control	7.1	7.4

Table 3: Coefficient of Variation between Tissue Replicates

	3 minute	1 hour
Negative Control	2.1	5.5
Octopamine hydrochloride	13	6.8
Positive control	29	14

CV (%) = 100 - [(lowest OD570/highest OD570) x 100%]

Applicant's summary and conclusion

Interpretation of results:

GHS criteria not met

Conclusions:

In conclusion, Octopamine hydrochloride is not 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 Octopamine hydrochloride for its ability to induce

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

possible corrosive potential of Octopamine hydrochloride 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 D151-1710037 of Octopamine hydrochloride was a white to off white powder.  Skin

tissue was moistened with 25 µL of Milli-Q water and at least 25 mg of Octopamine

hydrochloride was applied directly on top of the skin tissue.

The positive control had a mean relative tissue viability of 7.4% 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 <=13%, 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 Octopamine

hydrochloride compared to the negative control tissues was 94% and 60%, respectively.

Because the mean relative tissue viability for Octopamine hydrochloride was not below 50%

after the 3-minute treatment and not below 15% after the 1-hour treatment Octopamine

hydrochloride is considered to be not corrosive.

In conclusion, Octopamine hydrochloride is not corrosive in the in vitro skin corrosion test

under the experimental conditions described in this report.