Calcium-Silicium-Magnesium-Aluminium-Iron-Manganese oxide equivalent

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2009

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study performed according to GLP

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2009

Report date:

2009

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: ICCVAm Test Method Evaluation Report: Appendix G ICCVAM recommended HET-CAM Method Protocol (Nov. 2006)

Deviations:

no

Principles of method if other than guideline:

Background Review Document Current Status of In Vitro Test Methods for Identifying Ocular Corrosives and Severe Irritants: Hen’s Egg Test - Chorioallantoic Membrane Test Method (March 2006, NIH Publication No. 06-4515)
OECD 405 Draft Guideline “Acute Eye Irritation/Corrosion”, adopted 24. April 2002

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S)

IUPAC Name:

Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S)

Constituent 2

Reference substance name:

Slags, steelmaking, elec. furnace

EC Number:

294-410-9

EC Name:

Slags, steelmaking, elec. furnace

Cas Number:

91722-10-0

Molecular formula:

not applicable because slags are UVCB

IUPAC Name:

294-410-9

Details on test material:

Name Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S)
Appearance solid
Composition UVCB
CAS-No. 65996-69-2
EINECS-No. 266-002-0
Molecular formula not applicable (UVCB)
Molecular weight not applicable (UVCB
Purity 100 w/w % slag
Homogeneity not stated
Volatility extremely low (melting point > 300 °C)
Stability solid slag is stable at room temperature
Solubility slightly soluble in water
Date of production not stated
Date of expiry Year 2024
Storage conditions Room temperature 20 ± 5°C

Test animals / tissue source

Species:

other: hen

Strain:

other: Lohmann selected Leghorn chicken’s eggs

Details on test animals or tissues and environmental conditions:

in vitro test system HETCAM applied
Freshly laid Lohmann Leghorn chicken eggs were obtained from the LSL Rhine Main Gefluegelvermehrungsbetrieb (Dieburg, Germany) nine days before the start of the test. The eggs were incubated at 38.3 ± 0.2 °C and 58 % ± 2 % humidity in a still air incubator for eight days. During incubation, the eggs were rotated to prevent an attachment of the embryo to one side of the egg.

Test system

Vehicle:

unchanged (no vehicle)

Controls:

not required

Amount / concentration applied:

On the day of the test, the air bubble of a freshly incubated egg was marked. The egg was opened on the air bubble and the inner membrane was moistened with 0.9 % sodium chloride. After 30 min in the incubator, the solution was decanted and the inner membrane (CAM) was carefully removed with a forceps.
300 µL (equalling to 252.5 mg; this is the mean of all slag weights) of the test material were given directly onto the CAM surface in such a manner that 50% of the surface of the membrane were covered with test item.

Duration of treatment / exposure:

The reactions of the CAM were observed during 5 min:
• haemorrhage (bleeding from the vessels)
• vascular lysis (blood vessel disintegration)
• coagulation (intra- and extra-vascular protein denaturation)
The time for the appearance of each of the noted endpoints was monitored and recorded in seconds

Observation period (in vivo):

5 min

Number of animals or in vitro replicates:

0

Details on study design:

Sodium dodecyl sulfate solution C12H25NaO4S, CAS-No. 151-21-3. Solvent is deionised water, 1% solution (10 g/L) and
sodium hydroxide solution 0.1 n NaOH, CAS-No. 1310-73-2, dissolved in deionised water at 4 g/L, were used as positive controls:
Sodium chloride solution, NaCl, 0.9% solution in deionised water, was used as a negative control:
The following instruments and devices were used:
• Autoclave MLS 3020
• Candling light (for determination of the position of the egg bubble)
• Dentist’s rotating saw blade (for opening the egg)
• Tweezers
• Stop clock
• Scales Mettler PB 5001-S02
• Analytical scales MT AB 184 SA
• Heating chamber
• Orbital shaker GFL 3005
• Adjustable pipettes with sterile tips, LAUS No. 8 (0.2 – 2 mL)
• Digital Hygrometer
• Schott-bottles, glass vials, and culture flasks for solutions and media (sterilised before use by heating to 180 °C (two hours) or autoclavation)

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

The HET-CAM test

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Results

Results 1

Irritation parameter:

other: haemorrhage

Basis:

mean

Time point:

other: 5 min

Score:

0

Max. score:

0

Reversibility:

other: not relevant as no effect occured

Results 2

Irritation parameter:

other: lysis

Basis:

mean

Time point:

other: 5 min

Score:

0

Max. score:

0

Reversibility:

other: not relevant as no effect occured

Results 3

Irritation parameter:

other: coagulation

Basis:

mean

Time point:

other: 5 min

Score:

0

Max. score:

0

Reversibility:

other: not relevant as no effect occured

Irritant / corrosive response data:

see Table 3

Other effects:

no relevant effect

Any other information on results incl. tables

The observations which were made are given in the following table:

Table 3.  Experimental Observations

Test Group	Amount applied	Time until haemorrhage	Time until lysis	Time until coagulation
		seconds	seconds	seconds
Negative Control	300 µL	301	301	301
Negative Control	300 µL	301	301	301
Negative Control	300 µL	301	301	301
Positive Control 0.1-N NaOH	300 µL	8	34	14
Positive Control 0.1-N NaOH	300 µL	7	32	16
Positive Control 0.1-N NaOH	300 µL	7	29	17
Positive Control 1% SDS	300 µL	20	57	301
Positive Control 1% SDS	300 µL	17	58	301
Positive Control 1% SDS	300 µL	10	59	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	264.1 mg	301	301	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	250.7 mg	301	301	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	247.5 mg	301	301	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	240.8 mg	301	301	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	245.3 mg	301	301	301
Slag, ferrous metal, blast furnace (air cooled - ABS), fine-ground	266.3 mg	301	301	301

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: EU

Conclusions:

Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S) have no irritation potential

Executive summary:

Fine ground test material of slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S) was brought onto the surface of the chorio-allantoic membrane (CAM) of a hen’s egg which had been incubated at 38.3^oC and 58 % humidity for nine days.Observation time was five minutes at room temperature (negative controls: physiological sodium chloride solution, positive controls sodium dodecylsulfat (SDS, 1 %) and NaOH (0.1 n))

Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S) caused no effects on the blood vessels of the CAM (no haemorrhage, no lysis, no coagulation). The calculated mean irritation score is 0.0.

Slags, steelmaking, elec. furnace (stainless/high alloy steel production - EAF S) have no irritation potential.