Registration Dossier

Administrative data

Key value for chemical safety assessment

Genetic toxicity in vitro

Description of key information

Triallylcyanurate does not cause genotoxic effects in in vitro studies.

Link to relevant study records

Referenceopen allclose all

Endpoint:
in vitro cytogenicity / chromosome aberration study in mammalian cells
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-report according to guideline.
Qualifier:
according to
Guideline:
OECD Guideline 473 (In Vitro Mammalian Chromosome Aberration Test)
Qualifier:
according to
Guideline:
EU Method B.10 (Mutagenicity - In Vitro Mammalian Chromosome Aberration Test)
GLP compliance:
yes
Type of assay:
in vitro mammalian chromosome aberration test
Target gene:
not applicable
Species / strain / cell type:
lymphocytes: human, healthy donors without medication
Metabolic activation:
with and without
Metabolic activation system:
liver S9-mix of Aroclor 1254 induced rats
Test concentrations with justification for top dose:
0, 31.25, 62.5, 125, 250, 500 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen according to its solubility properties and its non-toxicity for the cells.
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO 1% (v/v)
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
mitomycin C
Remarks:
without metabolic activation

Migrated to IUCLID6: 0.1 or 0.2 µg/mL
Untreated negative controls:
no
Negative solvent / vehicle controls:
yes
Remarks:
DMSO 1% (v/v)
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
cyclophosphamide
Remarks:
with metabolic activation

Migrated to IUCLID6: 10 or 20 µg/ mL
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium

DURATION
- Preincubation period: 48 h
- Exposure duration: 4 h or 24 h without metabolic activation, 4 h with metabolic activation
- Fixation time (start of exposure up to fixation or harvest of cells): 22 h



SPINDLE INHIBITOR (cytogenetic assays): colcemid 0.9 µg/mL
STAIN (for cytogenetic assays): Giemsa stain


NUMBER OF REPLICATIONS: two cultures each two slide preparations


NUMBER OF CELLS EVALUATED: 100 metaphase per slide were evaluated


DETERMINATION OF CYTOTOXICITY
- Method: mitotic index


OTHER EXAMINATIONS:
- Determination of polyploidy: yes
Evaluation criteria:
Comparison of the number of chromosome aberrations of the samples with those of the solvent control. Total number of cells with aberrations exclusive of gap damage are analysed.
evaluation criteria:
The test is regarded to have a positive result, if the number of chromosomal aberrations is significantly increased (p<=0.05) compared with the solvent control, this increase is dose-dependent and both duplicate cultures lead to the same results. The increase should not occur in the severly cytotoxic range (mitotic index < 0.25).
Statistics:
Fisher-test (p<= 0.05)
Species / strain:
lymphocytes: human, healthy donors without medication
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
125 µg/ mL 24 h exposure, 250 µg/ mL 4 h exposure
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Remarks on result:
other: other: human peripheral lymphocytes
Remarks:
Migrated from field 'Test system'.

Treatment

µg/mL

S9-mix

4 h exposure

24 h exposure

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

DMSO

-

-

1.00

200

1.5

1.00

200

2.0

test substance

31.25

-

-

-

-

0.86

200

2.5

62.5

-

0.92

200

1.5

0.55

200

3.5

125

-

1.27

200

1.5

0.47

164#

3.7

250

-

0.58

183#

4.5

0.28

35#

3.2

500

-

0.43

101#

5.2

-

-

-

Mitomycin C

0.2

-

0.62

200

12.0*

0.55

200

13.0*

the following concentrations were not evaluated:

31.25 µg/ mL (4 h exposure), 500 µg/ mL ( 24 h exposure) 0.1 µg mitomycin C (4 and 24 h exposure)

Treatment

µg/mL

S9-mix

4 h exposure

Mitotic index#

number of metaphases scored

% of cells with aberrations excluding gaps

DMSO

-

+

1.00

200

1.0

test substance

31.25

+

-

-

-

62.5

+

0.77

200

2.0

125

+

1.62

200

1.0

250

+

0.40

183#

4.5

500

+

0.13

11#

0.0

Cyclophosphamide

10

+

0.63

200

10.5*

the following concentrations were not evaluated:

31.25 µg/ mL, 20 µg/ mL cyclophosphamide

#          mitotic index: number of metaphases/ 1000 cells: negative control = 1.00
##        no more metaphase of sufficient quality for evaluation due to cytotoxicity of Triallylcyanurate
*          significantly different from negative control (p=0.05)

Conclusions:
Interpretation of results (migrated information):
negative
Endpoint:
in vitro gene mutation study in bacteria
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-guideline study
Qualifier:
according to
Guideline:
OECD Guideline 471 (Bacterial Reverse Mutation Assay)
Version / remarks:
; 1983
Qualifier:
according to
Guideline:
EU Method B.13/14 (Mutagenicity - Reverse Mutation Test Using Bacteria)
Version / remarks:
; 1983
GLP compliance:
yes
Type of assay:
bacterial reverse mutation assay
Target gene:
His C3076, His D3052 or His G46
Species / strain / cell type:
S. typhimurium, other: Salmonella typhimurium strains TA 98, TA 100, TA 1535, TA 1537, TA 1538
Metabolic activation:
with and without
Metabolic activation system:
liver S9-mix of Aroclor 1254 induced male Wistar rats
Test concentrations with justification for top dose:
up to 2.5 mg/plate

remark:
In a preliminary cytotoxicity test where 0, 0.0005, 0.005, 0.05, 0.5, 5 or 50 mg/plate were applied, slight cytotoxicity was observed using 0.5 mg plate in strains TA 1537 and TA 100. Precipitation and evident cytotoxicity were observed in all test strains at 5 mg/plate.
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- test solution
- first assay: 0, 0.123, 0.37, 1.11, 3.33 and 10 mg/mL DMSO
- second assay: 0, 0.31, 0.93, 2.78, 8.33 and 25 mg/mL DMSO
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
sodium azide
Remarks:
without S9-mix

Migrated to IUCLID6: 1 µg per plate TA 1535 and TA 100
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
2-nitrofluorene
Remarks:
without S9-mix

Migrated to IUCLID6: 2 µg per plate TA1538, TA 98
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
9-aminoacridine
Remarks:
without S9-mix

Migrated to IUCLID6: 80 µg per plate TA 1537
Untreated negative controls:
not specified
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Remarks:
2 µg per plate TA 1535, TA 1538, TA 98, TA 100; 5 µg per plate TA 1537
Positive control substance:
other: 2-aminoanthracene
Remarks:
with S9-mix
Details on test system and experimental conditions:
IUCLID4 Type: Ames test
METHOD OF APPLICATION: in agar (plate incorporation);

DURATION
- Exposure duration: 3 d
- Selection time (if incubation with a selection agent): 3 d

SELECTION AGENT (mutation assays): histidine deficient agar

NUMBER OF REPLICATIONS: triplicates; two assays

DETERMINATION OF CYTOTOXICITY
- Method:relative total growth prior to mutagenicity experiment with up to 50 mg/ plate


Evaluation criteria:
a positive response in the assay system is taken to be a two-fold or greater increase in the mean number of revertant colonies appearing in the test plates over and above the background spontaneous reversion rate observed with the vehicle, together with evidence of a dose-response
Species / strain:
S. typhimurium TA 1535
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2.5 mg/plate with and without S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1537
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
0.833 mg/plate without S9-mix and 2.5 mg/plate with S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 1538
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
0.833 mg/plate without S9-mix and 2.5 mg/plate with S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 98
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
no cytotoxicity, but tested up to precipitating concentrations
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Species / strain:
S. typhimurium TA 100
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
2.5 mg/plate with and without S9
Vehicle controls validity:
valid
Untreated negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
precipitation occured at concentrations of 1 mg/ plate and more.
Remarks on result:
other: strain/cell type: S. typhimurium TA 1535
Remarks:
Migrated from field 'Test system'.
Conclusions:
Interpretation of results (migrated information):
negative
Endpoint:
in vitro gene mutation study in mammalian cells
Remarks:
Type of genotoxicity: gene mutation
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
other: GLP-Guideline study with acceptable restrictions: since the test was performed according to a previous guideline, only 1000 cells (instead of 2000 as required for the recent version of the guideline) were assessed.
Qualifier:
according to
Guideline:
EU Method B.17 (Mutagenicity - In Vitro Mammalian Cell Gene Mutation Test)
Version / remarks:
Cited as Directive 87/302/EEC, part B, p. 61
Qualifier:
according to
Guideline:
OECD Guideline 476 (In Vitro Mammalian Cell Gene Mutation Test)
GLP compliance:
yes (incl. certificate)
Remarks:
Hessisches Ministerium für Umwelt, Energie, Jugend, Familie und Gesundheit
Type of assay:
mammalian cell gene mutation assay
Target gene:
hypoxanthine-guanine phosphoribosyl transferase
Species / strain / cell type:
Chinese hamster lung fibroblasts (V79)
Details on mammalian cell type (if applicable):
- Type and identity of media: minimal essential medium (SEROMED, Berlin, Germany) with 10% FCS
- Properly maintained: yes
- Periodically checked for Mycoplasma contamination: yes
- Periodically checked for karyotype stability: yes
- Periodically "cleansed" against high spontaneous background: yes
Metabolic activation:
with and without
Metabolic activation system:
liver S9-mix of Aroclor 1254 induced male Wistar rats
Test concentrations with justification for top dose:
experiment I : 1, 3, 10, 20, 50, 80, 100, 200 or 300 µg/mL
experiment II : 5, 50, 100, 150, 180, 200, 220 or 250 µg/mL
Vehicle / solvent:
- Vehicle(s)/solvent(s) used: DMSO
- Justification for choice of solvent/vehicle: The solvent was chosen according to its solubility properties and its non-toxicity for the cells. Final DMSO-concentration in the culture medium did not exceed 1 %.
Untreated negative controls:
yes
Remarks:
untreated cells
Negative solvent / vehicle controls:
yes
Remarks:
DMSO
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
ethylmethanesulphonate
Remarks:
0.6 mg/ mL

Migrated to IUCLID6: without metabolic activation
Untreated negative controls:
yes
Remarks:
untreated cells
Negative solvent / vehicle controls:
yes
True negative controls:
not specified
Positive controls:
yes
Positive control substance:
7,12-dimethylbenzanthracene
Remarks:
3.85 µg/mL

Migrated to IUCLID6: with metabolic activation
Details on test system and experimental conditions:
METHOD OF APPLICATION: in medium


DURATION
- Preincubation period: 1 d
- Exposure duration: 4 h
- Expression time (cells in growth medium): 7 d
- Selection time (if incubation with a selection agent): 9 d
- Fixation time (start of exposure up to fixation or harvest of cells): 17 d


SELECTION AGENT (mutation assays): 11µg/ mL thioguanine
STAIN (for cytogenetic assays): methylene blue (10%) in 0.01% KOH solution


NUMBER OF REPLICATIONS: five replications, two independent experiments


NUMBER OF CELLS EVALUATED: about 3-5 x 10^5 cells/flask, stained colonies with more than 50 cells were counted


DETERMINATION OF CYTOTOXICITY
- Method: cloning efficiency, duplicates, two independent experiments
Evaluation criteria:
the gene mutation assay is considered acceptable if it meets the following criteria:
1. the number of mutant colonies per 10^6 cells found in the negative and/or solvent controls fall within the laboratory historical control data range: 0-45 mutants/ 10^6 cells
2. the positive control substance must produce a significant increase in mutant colony frequencies
3. The cloning efficiency (absolute value) of the negative and/or solvent controls must exceed 50%

A test substance is classified as mutagenic if it induces a reproducible mutation frequency that is at least three times higher than the spontaneous mutation frequency in the experiment at one or more of the concentrations. The test substance is classified as mutagenic if there is a reproducible concentration-related increase of the mutation frequency. Such evaluation may be considered also in the case that a threefold increase of the mutant frequency is not observed. However, in a case by case evaluation this decision depends on the level of the corresponding and historical negative control data.
Statistics:
since the distribution of mutant cells does not follow known statistical models, an adequate statistical method is not available
Species / strain:
Chinese hamster lung fibroblasts (V79)
Metabolic activation:
with and without
Genotoxicity:
negative
Cytotoxicity / choice of top concentrations:
cytotoxicity
Remarks:
100 µg/mL without S9-mix, 200 µg/mL with S9-mix
Vehicle controls validity:
valid
Untreated negative controls validity:
valid
Positive controls validity:
valid
Additional information on results:



RANGE-FINDING/SCREENING STUDIES: yes, XTT-Assay determining cytotoxicity up to 2.5 mg/mL tested. Here cytotoxicity was evident at concentrations higher than 100 µg/mL without S9-mix and 300 µg/mL with S9-mix.

The cloning efficiency of the cells was reduced to approx. 35 % without and approx. 26 % with metabolic activation at the highest evaluated concentration.

Remarks on result:
other: strain/cell type: Chinese hamster lung fibroblasts (V79)
Remarks:
Migrated from field 'Test system'.

Experiment I
(mutation rates)

c/ mL

S9-Mix

numbers of mutant colonies/ flask* found after plating in TG medium

mean (A,B,C,D,E)

S.D.

cell survival after plating in TG medium

mutant colonies /10^6 cells**

A

B

C

D

E

negative control

0

-

3

4

3

3

5

3.6

0.9

335272

10.7

solvent control DMSO

0

-

6

0

4

1

3

2.8

2.4

371280

7.5

positive control EMS

0.6

-

129

123

97

100

120

113.8

14.4

365366

311.5

test substance

1

-

culture was not continued

test substance

3

-

0

0

1

0

0

0.2

0.4

351879

0.6

test substance

10

-

culture was not continued

test substance

20

-

0

1

1

0

1

0.6

0.5

347809

1.7

test substance

50

-

2

2

0

5

1

2.0

1.9

350195

5.7

test substance

80

-

3

1

1

3

2

2.0

1.0

360000

5.6

negative control

0

+

1

3

3

1

0

1.6

1.3

323112

5.0

solvent control DMSO

0

+

2

0

3

0

2

1.4

1.3

349956

4.0

positive control DMBA

3.85

+

128

126

125

135

128

128.4

3.9

264148

486.1

test substance

1

+

5

2

2

4

4

3.4

1.3

365147

9.3

test substance

10

+

culture was not continued

test substance

50

+

5

5

3

4

6

4.6

1.1

300561

15.3

test substance

100

+

6

4

6

6

4

5.2

1.1

336894

15.4

test substance

200

+

4

1

6

4

2

3.4

1.9

270902

12.6

test substance

300

+

culture was not continued

Experiment II
(mutation rates)

c/ mL

S9-Mix

numbers of mutant colonies/ flask* found after plating in TG medium

mean (A,B,C,D,E)

S.D.

cell survival after plating in TG medium

mutant colonies /10^6 cells**

A

B

C

D

E

negative control

0

-

2

4

7

4

4

4.2

1.6

295557

14.2

solvent control DMSO

0

-

1

4

1

3

3

2.4

1.3

295581

8.1

positive control EMS

0.6

-

125

114

125

114

113

118.2

6.2

258016

458.1

test substance

5

-

5

9

5

6

3

5.6

2.2

308325

18.2

test substance

50

-

4

4

6

2

2

3.6

1.7

289038

12.5

test substance

100

-

2

3

2

1

3

2.2

0.8

325615

6.8

test substance

150

-

1

4

1

2

4

2.4

1.5

282530

8.5

test substance

180

-

culture was not continued

test substance

200

-

culture was not continued

negative control

0

+

2

4

2

7

4

3.8

2.0

238641

15.9

solvent control DMSO

0

+

2

1

1

2

2

1.6

0.5

294959

5.4

positive control DMBA

3.85

+

95

110

93

101

106

101.0

7.2

186956

540.2

test substance

5

+

2

1

1

0

1

1.0

0.7

243616

4.1

test substance

50

+

2

5

1

5

5

3.6

1.9

278924

12.9

test substance

100

+

1

4

0

2

1

1.6

1.5

323235

4.9

test substance

200

+

2

5

2

4

5

3.6

1.5

274105

13.1

test substance

220

+

culture was not continued

test substance

250

+

culture was not continued

*    only colonies with more than 50 cells 7 days after seeding were scored   
**  mean mutant colonies in TG medium* 10^6/ cell survival in TG medium 

In the first experiment with metabolic activation, the number of revertants was increased to 15.4 mutant colonies per 10^6 cells at a concentration of 100 µg/mL, compared to 4.0 spontaneaous revertant colonies of the corresponding solvent control. Although a factor of three required to be judged as a relevant increase was exceeded, the absolute value is well within the range of historical controls. Furthermore, there was no indication of a concentration dependent increase of mutant colonies and the effect is considered to be due to the low negative and solvent control levels. In the second experiment there was no relevant increase of mutant colony numbers at any concentration level, neither with or without metabolic activation.

Conclusions:
Interpretation of results (migrated information):
negative

Genetic toxicity in vivo

Description of key information

Triallylcyanurate does not cause cytogenic effects in the mouse micronucleus test.

Link to relevant study records
Reference
Endpoint:
in vivo mammalian somatic cell study: cytogenicity / erythrocyte micronucleus
Remarks:
Type of genotoxicity: chromosome aberration
Type of information:
experimental study
Adequacy of study:
key study
Reliability:
1 (reliable without restriction)
Rationale for reliability incl. deficiencies:
other: GLP-report according to guideline.
Qualifier:
according to
Guideline:
OECD Guideline 474 (Mammalian Erythrocyte Micronucleus Test)
Version / remarks:
; 1983
GLP compliance:
yes
Type of assay:
micronucleus assay
Species:
mouse
Strain:
NMRI
Sex:
male/female
Details on test animals and environmental conditions:
Source: Winkelmann, Borchen
Age at study initiation: 6 weeks
Weight at study initiation: males 21-27
females 21-30
Route of administration:
oral: gavage
Vehicle:
Peanut oil
Details on exposure:
Three groups of mice, the negative and positiv control groups (18 animals per sex) and the test material group
(21 animals per sex), each received a single administration by oral gavage (diet withdrawal: 16 h before treatment).
Group 1, the negative control, received peanut oil, group 2, the test material group, received 316 mg/kg body weight of the test material.
Group 3, the positive control, received cyclophosphamide (51.1 mg/kg body weight) dissolved in physiological saline solution (0.9 %).
Duration of treatment / exposure:
24, 48 or 72 hours
Frequency of treatment:
Single administration
Post exposure period:
No
Remarks:
Doses / Concentrations:
316 mg/kg
Basis:
nominal conc.
No. of animals per sex per dose:
control groups: 18
test group: 21
Control animals:
yes, concurrent vehicle
Positive control(s):
cyclophosphamide (51.1 mg/kg body weight) dissolved in physiological saline solution (0.9 % (w/w))
Tissues and cell types examined:
Bone marrow smears (at least two slides per animal) from the first 5 animals per sex and group were used for evaluation.
One slide per animal was examined. The remaining smears of each sex and group per interval were evaluated if macroscopical examination
of the first smears revealed technical imperfections which precluded aceurate microscopical analysis.
From each animal 1000 polychromatic erythrocytes (PCE) were scored for the incidence of micronuclei.
The ratio of polychromatic to normochromatic erythrocytes (PCE/NCE) was calculated, based on 1000 erythrocytes (PCE + NCE) scored per slide,
as a measure of the toxic efficacy of the test material.

Evaluation criteria:
If a test material produced no statistically significant and reproducible positive response at any one of the test points compared to the negative
control group, it was considered non-mutagenic.
Statistics:
Poisson test
Sex:
male/female
Genotoxicity:
negative
Toxicity:
yes
Vehicle controls validity:
valid
Negative controls validity:
not specified
Positive controls validity:
valid
Additional information on results:
Triallylcyanurate (TAC) related toxic symptoms were registered in all test material group animals. The symptoms consisted in coordination
disturbances, clonic convulsions, decrease of muscle tone, loss of righting reflexes, loss of pinna reflex, loss of pain reflex, loss of corneal reflex,
ptosis, tear formation, strenuous respiration, and reduced temperature of the body surface. One female animal of the test material group died.

Reduction in the ratio of polychromatic to normochromatic erythrocytes was present in all dose groups of the test material indicating a toxic effect 
on the bone marrow. No statistically significant test material-related increase in the number of micronucleated polychromatic erythrocytes was 
observed in either male or female animals and also if both sexes were analysed combined at the 48 hr and 72 hr sampling times and in females at the 24 hr sampling time. At the 24 hr sampling time a statistically significant increase in micronuclei was observed only in male animals of the main test. 

This increase could not be verified in the repetition test using two additional dose levels and is therefore considered an incidental finding. 
Conclusions:
Triallylcyanurate does not cause cytogenic effects in the mouse micronucleus test.

Additional information

Outcome of studies with triallyl cyanurate was negative in a bacterial reverse mutation assay (with and without metabolic activation), an in vitrogene mutation test (HPRT test) in Chinese hamster V79 cells (with and without metabolic activation) as well as an in vitro chromosomal aberration assay in human peripheral lymphocytes. The in vivo micronucleus assay in mice was also negative.


Short description of key information:
Not mutagenic in bacteria (Ames Test) with and without metabolic activation.
Not mutagenic in vitro in Chinese hamster lung fibroblasts (HPRT test).
Not clastogenic in vitro in human peripheral lymphocytes (Chromosome Aberration Test).
Negative in Mammalian Erythrocyte Micronucleus Test in vivo in mice.

Endpoint Conclusion: No adverse effect observed (negative)

Justification for classification or non-classification

No genotoxic potential is concluded for triallyl cyanurate and therefore no classification is needed according to DSD (67/548/EEC) and CLP (1272/2008/EC).