Ladle skimming slag, Ladle furnace refining, slag, Ferrous slag

Eye irritation

Administrative data

Endpoint:

eye irritation: in vitro / ex vivo

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2010-2011

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:

2011

Report date:

2011

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD-Guideline 437

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

Slags, steelmaking (SMS)

IUPAC Name:

Slags, steelmaking (SMS)

Constituent 2

Reference substance name:

Slag, pig iron ladle refining, non-granulated, water-cooled

Molecular formula:

not applicable because slags are UVCB

IUPAC Name:

Slag, pig iron ladle refining, non-granulated, water-cooled

Details on test material:

Name Slags, steelmaking - SMS, fine ground (< 90 µm)
Appearance grey granules (soldis)
Molecular formula not applicable (UVCB)
Molecular weight not applicable (UVCB)
Purity 100 w/w % slag
Homogeneity homogenous
Vapour pressure extremely low (melting point >300°C)
Stability solid slag is stable at room temperature
Solubility slightly soluble in water
Production date not stated (2009)
Expiry date 12/2024
Storage Room Temperature: (20 ± 5°C)

Test animals / tissue source

Species:

other: cow

Strain:

other: freshly bovine eyes from the slaughterhouse Müller Fleisch GmbH

Details on test animals or tissues and environmental conditions:

Freshly bovine eyes were obtained from the slaughterhouse Müller Fleisch GmbH, Enzstr. 2.4, 75217 Birkenfeld, Germany, on the day of the test. The cattle were between 12 and 60 months. The eyes were transported to the test facility in Hank’s balanced salt solution with 0.01% streptomycin and 0.01% penicillin. Then the corneas were dissected and incu-bated with media at 32 ± 1°C in an incubation chamber for 1 hour.

Test system

Vehicle:

water

Controls:

not required

Amount / concentration applied:

For each treatment group (negative control, positive control and test item), three replicates were used. 750 µl negative control resp. eluate of the test item resp. positive control solu-tion were applied to each replicate.

Duration of treatment / exposure:

Exposition time on the corneas was 10 min at 32°C. After thorough rinsing with cMEM with phenol red and final rinsing with cMEM without phenol red, the anterior chamber was filled with cMEM without phenol red, and the corneas were stored for an additional two hours at 32°C (post-incubation).

Number of animals or in vitro replicates:

0

Details on study design:

Chemicals
• Incubation Medium MEM (Minimum Essential Medium) without phenol red, Supplier Gibco/Invitrogen
• Incubation Medium MEM (Minimum Essential Medium) with phenol red, Supplier Gibco/Invitrogen
• Sodium bicarbonate NaHCO3, Supplier Merck
• L-glutamine, supplier: Biochrom AG
• 1% fetal calf serum, supplier PAN BioTech (EU approved)
• cMEM without phenol red: 495mL MEM are supplemented with 1.1 g NaHCO3, 5 mL FCS and 4.57 g L-Glutamine.
(As soon as the MEM without phenol red is supplemented with NaHCO3, FCS and L-Glutamine, it is designated in the following as cMEM ( = complete MEM) without phenol red.)
• cMEM with phenol red: 495 mL MEM are supplemented with 1.1 g NaHCO3, 5 mL FCS and 4.86 g L-Glutamine.
(As soon as the MEM with phenol red is supplemented with NaHCO3, FCS and L-Glutamine, it is designated in the following as cMEM ( = complete MEM) with phenol red.)
• Fluorescein-Na
• Hank’s Balanced Salt Solution (HBSS) without Phenol red, supplier: Life Technolo-gies
• 0.01% Penicillin-Streptomycin solution, supplier: PAN Biotech

Negative Control
• Sodium Chloride Solution: NaCl, CAS-No. 7647-14-5, 0.9% solution in deionised water

Results and discussion

In vitro

Results

Irritation parameter:

in vitro irritation score

Run / experiment:

BCOP Test

Negative controls validity:

valid

Positive controls validity:

valid

Remarks on result:

no indication of irritation

In vivo

Results

Irritation parameter:

other: lysis

Basis:

mean

Time point:

other: 10 minutes

Score:

0.956

Max. score:

0

Reversibility:

other: not relevant as no effect occured

Irritant / corrosive response data:

see Table 5

Other effects:

no relevant effect

Any other information on results incl. tables

In a test according to OECD TG 405 with solid material, irritation was observed. As was shown by SEM and other techniques, the test material was coarse and consisted of sharp-edged particles which apparently were the cause of the effect. The effect was fully reversible. In ISO 10993 "Biological Testing of Medicinal and Dental Materials and Devices Part 1: Guidance on Selection Tests" it is required that solid materials are tested as leachates.

Opacity and Permeability Values

The opacity values which were measured before and after exposition are given in the following table:

Table3     Opacity values

Test group	Absorption at 570 nm before exposition	Opacity before exposition	Absorption at 570 nm after exposition	Opacity after exposition	Opacity difference
Negative control 0.9% NaCl	0.2017	1.5912	0.2206	1.6617	0.0705
	0.2048	1.6026	0.2147	1.6395	0.0370
	0.2054	1.6049	0.1788	1.5094	-0.0954
10052603G	0.1821	1.5210	0.4262	2.6678	1.1468
	0.2160	1.6443	0.4208	2.6351	0.9908
	0.2366	1.7242	0.4267	2.6712	0.9470
Positive control 10% NaOH	0.1531	1.4228	2.0438	110.6193	109.1966
	0.2195	1.6575	2.2139	163.6342	161.9767
	0.1278	1.3420	2.1291	134.6087	133.2667

Mean opacity difference of the negative control is 0.0040.

For the permeability measurement, a twelve-fold measurement of a medium blank (cMEM without phenol red) was performed. The mean optical density of 0.042 was subtracted from all measurements.

Three replicates for each cornea were measured. The optical density values at 490 nm are given in the following table:

Table4     Optical density at 490 nm

Repl.	Negative control			10052603G			Positive control
1	0.047	0.047	0.043	0.046	0.037	0.047	1.306	1.284	2.108
2	0.044	0.047	0.042	0.044	0.036	0.045	1.287	1.301	2.099
3	0.046	0.048	0.043	0.046	0.038	0.044	1.291	1.277	2.097
Mean	0.046	0.047	0.043	0.045	0.037	0.045	1.295	1.287	2.101
Net	0.004	0.005	0.001	0.003	-0.005	0.003	1.253	1.245	2.059
Net – Neg. Contr.				0.000	-0.008	0.000	1.250	1.242	2.056

From all absorption means, the mean absorption of the medium blank (0.042) was subszracted, given the Net mean absorption. Then, the net mean absorption of the negative control (0.003) was substracted from the individual replicates of test item and positive control. Additionally, the path length of 1.695 was taken into account when calculating the IVIS.

IVIS Values The calculated IVIS for each replicate and the corresponding means are presented in the following table:

Table5     IVIS

Test group	IVIS	Mean IVIS	Relative Standard Deviation IVIS
Negative control 0,9% NaCl	0.1722	0.0890	154.6%
	0.1641
	-0.0700
10052603G	1.1428	0.9560	18.8%
	0.7834
	0.9430
Positive control 10% NaOH	140.9722	173.3520	16.3%
	193.5489
	185.5338

 

Note: the high relative standard deviation of the IVIS of test item and negative control is due to mathematical reasons, as the respective means are very small.

Applicant's summary and conclusion

Interpretation of results:

not irritating

Remarks:

Migrated information Criteria used for interpretation of results: OECD GHS

Conclusions:

Slags, steelmaking - SMS have no irritation potential

Executive summary:

One valid experiment was performed.

Bovine corneas were used. They were collected from slaughtered cattle which were between 12 and 60 months old.

The test item Slag, steelmaking - SMS, fine-ground (< 90 µm) was brought onto the cornea of a bovine eye which had been incubated with cMEM without Phenol red at 32±1°C for one hour and whose opacity had been measured. The test item was incubated on the cornea for 10 minutes at 32±1°C. After removal of the test item and two hours post-incubation, opacity and permeability values were measured.

Physiological sodium chloride solution was used as negative control. The negative control showed no irritating effect on the cornea.

10% sodium hydroxide solution was used as positive control. The positive control induced a very severe irritation on the cornea.

The test item Slag, steelmaking - SMS, fine-ground (< 90 µm) showed no effects on the cornea of the bovine eye. The calculated IVIS (in vitro irritancy score) is 0.956.

In conlcusion it can be stated, that in this study and under the experimental conditions reported, the test item Slag, steelmaking converter - BOS, fine ground (< 90 µm) possesses no eye irritation potential.