D-Gluconic acid, δ-lactone, reaction products with propanolamine

Endpoint summary

Key value for chemical safety assessment

Skin irritation / corrosion

Link to relevant study records

Reference

Endpoint:

skin irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 439 (In Vitro Skin Irritation: Reconstructed Human Epidermis Test Method)

Deviations:

no

GLP compliance:

no

Remarks:

The study was performed according to national and/or international guidelines applying a study plan and Standard Operating Procedures. Although the performing laboratory is not GLP certified, test procedures are generally following GLP guidance and rules.

Specific details on test material used for the study:

Chemical name: Samson
Code: S18+
CAS number: -
Batch number: -
Purity: not specified
Physical state: Liquid
Storage conditions: Room temperature

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:

Description: Ashland 0.5 cm2 reconstructed epidermis from normal human keratinocytes. Cells are grown on inert polycarbonate filter on chemically defined medium. airlifted for 17 days.
Batch N°: 1503EPID03
Origin: Foreskin
Quality Controls: Certified by Ashland Tissue Engineering laboratories. April 6, 2015

Control samples:

yes, concurrent negative control

yes, concurrent positive control

Amount/concentration applied:

100%

Duration of treatment / exposure:

42 minutes

Duration of post-treatment incubation (if applicable):

42 hours, duration of MTT incubation: 3 hours

Number of replicates:

9

Irritation / corrosion parameter:

% tissue viability

Run / experiment:

Viability (%)

Value:

>= 90.3 - <= 95.3

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

Viability 87 to 107%

Positive controls validity:

valid

Remarks:

Viability < 2%

Interpretation of results:

GHS criteria not met

Conclusions:

According to the In-Vitro Skin Irritation by the Ashland Epidermis Equivalent (Aee) Test Method reaction mass of hydroxylpropyl gluconamide and hydroxypropyl ammonium gluconate is classified as Non Irritant.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Eye irritation

Link to relevant study records

Reference

Reference 1

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

March 30 to April 9, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with national standard methods

Qualifier:

according to guideline

Guideline:

other: Hens Egg Test on the Chorio-Allantoic Membrane (HET-CAM).

Version / remarks:

Journal officiel de la République Française of December 26, 1996 with some modifications described in the Standard Operation Procedure N°TAM-01-v2

Deviations:

no

GLP compliance:

no

Remarks:

The study was performed according to national and/or international guidelines applying a study plan and Standard Operating Procedures. Although the performing laboratory is not GLP certified, test procedures are generally following GLP guidance and rules.

Specific details on test material used for the study:

ID: Samson 18+
Appearance: liquid
Storage: room temperature

Species:

chicken

Details on test animals or tissues and environmental conditions:

Eggs reception and incubation
- Upon receipt, the cracked or broken eggs are discarded. Before incubation, the eggs are stored in egg boxes (blunt ends upwards). If the incubation didn’t take place in few hours after reception, eggs are stored at 8°C; and kept away from light for a maximum of 7 days, then for 24 h at 20°C before being placed in an incubator.
- Clean and fertile White Leghorn chicken eggs are placed in an incubator for eight days with a rotating tray at 37.8 ± 0.5ºC and 52 ± 2% relative humidity.

Vehicle:

water

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

S18+ at 100%; 10%; 5% in 300 µL.

Duration of treatment / exposure:

300 seconds

Duration of post- treatment incubation (in vitro):

Not applicable

Number of animals or in vitro replicates:

Test article groups: 4; Controles: 2

Details on study design:

This test is carried out on 9 days’ old, fertilized and embryonated chicken eggs. The eggshell and inner membrane are carefully removed taking care that the CAM is not injured.
- Each test substance (if included, benchmark and solvent/vehicle controls) is applied to 4 eggs. Positive and negative controls are applied to only two eggs.
- 300 μl of test product are then deposited on the membrane. A timer is started immediately after application.
- At 20 seconds, test substance is removed from the membrane by rinsing the CAM with approximately 2x2.5 ml of DPBS 1X.
- The CAM reactions are observed over a period of 300 seconds. The time of the appearance of HAEMORRHAGE, HYPERAEMIA and COAGULATION should be monitored and recorded, in seconds
- Pictures are taken at 0, 30, 120 and 300 seconds and recorded as t=0s, t=30s, t=120s and t=300s for the corresponding tested substance.
- For each egg, the numerical time-dependant scores for HAEMORRHAGE, HYPERAEMIA and COAGULATION are summed to give a single numerical value.
- The Irritation Score (IS) of the tested product is the arithmetic average, rounded to the decimal, of the scores obtained on four eggs (treated with a same test substance). Therefore, the maximal score is 21.
IS < 1 = Practically not irritant
1 ≤ IS < 5 = Slightly irritant
5 ≤ IS < 9 = Moderately irritant
IS ≥ 9 = Irritant

Irritation parameter:

in vitro irritation score

Run / experiment:

100%

Value:

2.5

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

Value = 0

Positive controls validity:

valid

Remarks:

Values: NaOH 1N = 19; NaOH 0.1N= 10

Remarks on result:

other: Slightly irritant

Irritation parameter:

in vitro irritation score

Run / experiment:

10%

Value:

0

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

Interpretation of results:

GHS criteria not met

Conclusions:

According to HET-CAM test, Samson S18+ at 100% is identified as Slightly irritant and Samson S18+ at 10% in H2O is identified as Practically not irritant.

Reference 2

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

April, 2015

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

test procedure in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

equivalent or similar to guideline

Guideline:

OECD Guideline 492 (Reconstructed Human Cornea-like Epithelium (RhCE) Test Method for Identifying Chemicals Not Requiring Classification and Labelling for Eye Irritation or Serious Eye Damage)

Version / remarks:

- SKINETHIC Eye Irritation Test- Standard Operating Procedure – “SkinEthicTM Human Corneal Epithelial Model (SkinEthicTM HCE) Long exposure-time treatment”, 2010.
- N.Alépée et al., Cosmetics Europe multi-laboratory pre-validation of the SkinEthicTM reconstituted human corneal epithelium test method for the prediction of eye irritation, 2013.
- SKINETHIC website - http://www.skinethic.com/OC_IRRITATION_CHEM.asp OCULAR IRRITATION ASSAY, 2014

Deviations:

not specified

GLP compliance:

no

Remarks:

The study was performed according to national and/or international guidelines applying a study plan and Standard Operating Procedures. Although the performing laboratory is not GLP certified, test procedures are generally following GLP guidance and rules.

Specific details on test material used for the study:

Chemical name: Samson
Code: S18+
CAS number: -
Batch number: -
Purity: not specified
Physical state: Liquid
Storage conditions: Room temperature

Species:

human

Details on test animals or tissues and environmental conditions:

Ashland Reconstituted Human Corneal Epithelial Model
Description: Human Corneal Epithelium
Batch N°: 1504HCE01
Origin: Immortalized Human Corneal Epithelium cells
Quality Controls: April 13, 2015

Vehicle:

unchanged (no vehicle)

Controls:

yes, concurrent positive control

yes, concurrent negative control

Amount / concentration applied:

100%

Duration of treatment / exposure:

1 h

Duration of post- treatment incubation (in vitro):

16 h, MTT incubation: 3 h

Number of animals or in vitro replicates:

9

Details on study design:

EXPERIMENTAL PROTOCOL
1. First day: Pre-incubation step
- For a given chemical 3 tissues were be used.
- An appropriate numbers of 6-well plates were filled with 1 ml of maintenance culture medium.
- HCE tissues were transferred into maintenance medium filled wells, using sterile forceps.
- Place the HCE tissues at 37°C, 5% CO2 until test substance application (usually 24 hours).
2 Second day: Application / Rinsing
- 24-well plate was filled with 300μl by well of pre-warmed maintenance culture medium.
- Tissues were transferred.
- 30 μL ± 0.5 μL (or 30 μg) of products and controls were dispensed on the top of cornea.
- Plates containing the treated corneas were kept at room temperature for 1 hour.
- Tissues were rinsed 3 times with 5 mL of DPBS (adjust the distribution to 0.5 mL per push). Remaining DPBS was removed by tapping the inserts on absorbent paper.
- Epitheliums were transferred in a new 24-well plate containing 300 μl of fresh maintenance medium.
- Treated and rinsed corneas were incubated at 37°C, 5% CO2 for 16 hours (± 60 minutes).
3 Third day: Viability measurement
- 24-well plates were filled with 300 μL MTT solution (0.5mg/ml).
- Treated tissues were transferred in the pre-filled MTT 24-well plates and Incubated for 3 hours (± 5 minutes) at 37°C and 5% CO2.
- Treated tissue insert bottom was dried on sterile absorbent paper and transferred in new plate containing 750 μl of isopropanol solution.
- 750 μL isopropanol solutions were added on the top of each tissue.
- Plate was protected from evaporation by stretching 3 parafilm layers over the plate and adding the lid on the plate and incubated for 2 hours (± 5 minutes) at room temperature with gentle agitation (about 150 rpm) for formazan extraction.
- Tissue and polycarbonate filter were pierced with a tip in order to get the whole extraction solution in the corresponding well.
- Extraction solution was homogenized by pipetting 3 times up and down to complete formazan crystals solubilization.
- 2 x 200 μL extraction solution per well (= 2 wells per tissue i.e. 2 replicates per tissue) were transferred into a 96-well plate.
- Optical Densities (OD) was read using a 96-well plate spectrophotometer: The concentration of formazan was measured by determining the OD at 570 nm.

Irritation parameter:

in vitro irritation score

Run / experiment:

Viability (percentage)

Value:

98.4

Vehicle controls validity:

not applicable

Negative controls validity:

valid

Remarks:

100% viability

Positive controls validity:

valid

Remarks:

21.9 % viability

Interpretation of results:

GHS criteria not met

Conclusions:

According to Ashland Reconstituted Human Corneal Epithelial (RHCE) Long exposure-time treatment test, Samson S18+ displayed no irritation potential.

Endpoint conclusion

Endpoint conclusion:

no adverse effect observed (not irritating)

Respiratory irritation

Endpoint conclusion

Endpoint conclusion:

no study available

Additional information

Skin irritation:

According to the  In-Vitro Skin Irritation by the Ashland Epidermis Equivalent (Aee) Test Method reaction mass of hydroxylpropyl gluconamide and  hydroxypropyl ammonium gluconate is classified as Non Irritant.

Eye irritation:

According to a Reconstituted Human Corneal Epithelial (RHCE) Long exposure-time treatment test, the reaction mass displayed no irritation potential. According to HET-CAM test, the reaction mass is identified as Slightly irritant at 100% and as Practically not irritant at 10% in H2O.

Justification for classification or non-classification

Based on the in-vitro studies performed according to internationally accepted protocols, Reaction mass of 3-hydroxypropan-1-aminium D-gluconate and N-(3-hydroxypropyl)-D-gluconamide does not meet the criteria for classification as irritant for skin or eye.