Reaction mass of lithium sodium 5-amino-3-{[4-(2-{4-[(7-amino-1-hydroxy-3-sulfo-2-naphthyl)diazenyl]-2-sulfophenyl}vinyl)-3-sulfophenyl]diazenyl}-4-hydroxynaphthalene-2,7-disulfonate 2,2'-(methylimino)diethanol (1:1) and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[6-amino-4-hydroxynaphthalene-2-sulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol and 3,3'-[vinylenebis[(3-sulpho-p-phenylene)azo]]bis[5-amino-4-hydroxynaphthalene-2,7-disulphonic] acid, lithium sodium salt, compound with 2,2'-(methylimino)diethanol

Toxicological Summary

Workers - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

17.63 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

30

Modified dose descriptor starting point:

NOAEC

Value:

528.947 mg/m³

Explanation for the modification of the dose descriptor starting point:

NOAEL(rat) subacute oral toxicity study: 300 mg/kg bw/day Correction of the starting point according to ECHA Guidance: There are no quantitative data on absorption rates for oral or inhalation route available. Based on the available systemic toxicity following oral application, log Pow in the range of 1.82 - 3.72 and high solubility in water absorption by the respiratory tract is likely and can be assumed to be in the same range than oral absorption. Therefore there is no need to introduce further default values for extrapolation from oral route to inhalation route. Corrected inhalatory NOAEC = NOAEL(oral x 1/0.38 x 6.7/10 x 1 Corrected NOAEC (worker) = 528.947 mg/m³.

AF for dose response relationship:

1

Justification:

The NOAEL is derived from a GLP study according to OFCD TG 407 and was evaluated with Klimisch sore 1. Consequently to this high quality study which complies with the requirements of today ECHA proposes a default assessment factor of 1 (R8, p30)

AF for differences in duration of exposure:

6

Justification:

Sub-acute to chronic exposure

AF for interspecies differences (allometric scaling):

1

Justification:

Allometric scaling rat versus human: the AF of 4 is already included in the route to route extrapolation

AF for other interspecies differences:

1

Justification:

In an evaluation by ECETOC 2003 and 2010 it is considered that routine application of the factor 2.5 is scientifically unjustified as a default factor.

AF for intraspecies differences:

5

Justification:

The NOAEL is not the limit dose and absorption of the test substance can be assumed as demonstrated by log Pow and solubility in water although the molecular weight of Bayscript Blaukomponente cannot be given. Thus the standard default factor as suggested by ECHA is applied

AF for the quality of the whole database:

1

Justification:

There is information available to cover all relevant toxicological endpoints. The available studies are at most performed according to guideline and GLP.

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

35.26 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

DNEL extrapolated from long term DNEL

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

Workers - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

2.5 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

120

Modified dose descriptor starting point:

NOAEL

Value:

300 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

There is no long term dermal study available. Therefore, the oral NOAEL of the sub-acute toxicity study has to be considered instead. According to ECHA Guidance Document, Chapter R8 Version 2, December 20120, no additional default factors have to be introduced when performing oral-to-dermal extrapolation because it can be assumed that dermal absorption will not be higher than oral absorption. NOAEL (oral) = NOAEL (dermal) = 300 mg/kg bw/day.

AF for dose response relationship:

1

Justification:

The NOAEL is derived from a GLP study according to OFCD TG 407 and was evaluated with Klimisch sore 1. Consequently to this high quality study which complies with the requirements of today ECHA proposes a default assessment factor of 1 (R8, p30)

AF for differences in duration of exposure:

6

Justification:

Sub-acute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

Allometric scaling rat versus human

AF for other interspecies differences:

1

Justification:

In an evaluation by ECETOC 2003 and 2010 it is considered that routine application of the factor 2.5 as a default factor is scientifically unjustified.

AF for intraspecies differences:

5

Justification:

The NOAEL is not the limit dose and absorption of the test substance can be assumed as demonstrated by log Pow and solubility in water although the molecular weight of Bayscript Blaukomponente cannot be given. Thus the standard default factor as suggested by ECHA is applied.

AF for the quality of the whole database:

1

Justification:

There is information available to cover all relevant toxicological endpoints. The available studies are at most performed according to guideline and GLP.

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

5 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

DNEL extrapolated from long term DNEL

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

Workers - Hazard for the eyes

Local effects

Hazard assessment conclusion:

no hazard identified

Additional information - workers

Bayscript Blaukomponente

EC-Nr. 916 -916 -7

DNEL-Derivation: Worker

I. Introduction

Bayscript Blaukomponente is not classified legally. There is no German or European OEL available which could be taken as starting point for DNEL derivation.

II. Derivation of DNELs systemic

Basis for DNEL-Derivation

Bayscript Blaukomponente (konserviert) was administered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle, in daily doses of 0, 100, 300, 1000 mg/kg bw/day for a period of four weeks. This GLP study was performed according to OECD guideline 407.

Treatment with Bayscript Blaukomponente resulted in several findings which were related to the color of the test substance. The whole body and several organs of animals treated with 1000 mg/kg bw/day appeared discolored. The kidneys were also discolored in the lower dose groups. In the kidneys, beneath findings associated with the color of the test substance (starting at 300 mg/kg b.w./day), corticotubular degeneration and increased incidence and/or severity of cortical basophilic (regenerative) tubules were noted at 1000 mg/kg b.w./day. Furthermore, demyelinated fibers in nerve roots at the spinal cord were seen in both sexes at 1000 mg/kg b.w./day. At 1000 mg/kg b.w./day, histopathological findings in the mesenteric lymph node, the interstitium of the testis, and in the Peyer's patch were related to the colored test item and not accompanied by other structural changes of the organs.

Minor changes in red blood cell parameters observed for males and females at 1000 mg/kg b.w./day were associated with appearing of golden-brown pigment deposition in the spleen in females. In males, the thrombocyte count was increased at 1000 mg/kg b.w./day.

Under the conditions described the no adverse observed effect level (NOAEL) for administration of BAYSCRIPT Blaukomponente (konserviert) to male and female Wistar rats was 300 mg/kg b.w./day mainly due to degenerative effects on kidneys and nerve roots.

1.)Worker DNEL (long term, inhalation route systemic effects)

NOAEL(rat) subacute oral toxicity study: 300 mg/kg bw/day

Correction of the starting point according to ECHA Guidance:

There are no quantitative data on absorption rates for oral or inhalation route available. Based on the available systemic toxicity following oral application, log Pow in the range of 1.82 - 3.72 and high solubility in water absorption by the respiratory tract is likely and can be assumed to be in the same range than oral absorption. Therefore there is no need to introduce further default values for extrapolation from oral route to inhalation route.

Corrected inhalatory NOAEC = NOAEL(oral x 1/0.38 x 6.7/10 x 1

Corrected NOAEC (worker) = 528.947 mg/m³

Overall factor30

Worker DNEL (long term, systemic for inhalation route)17.63 mg/m³

2.)DNEL short term inhalation route – Systemic effects

There are no data on acute inhalation toxicity available. As acute effects may manifest themselves not immediately but also in considerable time after the exposure ECHA proposes in the Guidance Document R8, 2012, that for short term inhalation exposure the long-term DNEL should be used as starting point and a factor of 2 should be applied to extrapolate from long term DNEL to short term DNEL based on available animal data.

Thus, Worker DNEL (short term, systemic for inhalation exposure) accounts for 35.26 mg/m³

3.)DNEL (long term, dermal route – systemic effects)

There is no long term dermal study available. Therefore, the oral NOAEL of the sub-acute toxicity study has to be considered instead. According to ECHA Guidance Document, Chapter R8 Version 2, December 20120, no additional default factors have to be introduced when performing oral-to-dermal extrapolation because it can be assumed that dermal absorption will not be higher than oral absorption

NOAEL (oral) = NOAEL (dermal) = 300 mg/kg bw/day

Overall factor120

Worker DNEL (long term, systemic for dermal route) 2.5 mg/kg bw/day

4.)DNEL (short term, dermal route - systemic effects)

There are acute toxicity studies available using the dermal exposure route. These studies comprise acute toxicity, skin irritation and sensitization. In none of these three study types mortality or signs of intoxication were observed. The LD50 (dermal) is greater 2000 mg/kg bw and the test substance is not irritating to the skin nor caused skin sensitization. As the NOAEL for acute dermal toxicity is not determined exactly it is proposed to take the derived DNEL (long term, systemic, dermal route) as starting point for a DNEL(short term, systemic, dermal route). According to ECHA Guidance Document R8 a factor of 2 should be applied based on the available animal data.

Worker- DNEL (short term, systemic for dermal route) = 5 mg/kg bw/day

.

III. Derivation of DNELs (local)

Basis for derivation DNELs local

The available studies on skin and eye irritation are performed according to OECD TG 404 and 405, respectively, under GLP conditions. Bayscript Blaukomponente was not irritating to the skin of rabbits and was not irritating to the mucous membranes of the eyes of rabbits. The modified Local Lymph Node Assay (IMDS) was performed according to OECD Guidelines No. 429 and No. 406. The results show that there is no indication for skin sensitizing effect after administration of Bayscript Blaukomponente.

Based on these results no DNELs (local) need to be derived because no hazard was identified.

IV. Conclusion (systemic and local)

The relevant DNELs are

Long-term: Worker DNEL (systemic inhalation route): 17.63 mg/m³

Long-term: Worker DNEL (systemic, dermal route): 2.5 mg/kg bw/day

Short-term Worker DNEL (systemic inhalation route): 35.26 mg/m³

Short-term Worker DNEL (systemic dermal route): 5 mg/kg bw/day

No local hazard was identified.

General Population - Hazard via inhalation route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

4.35 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

60

Modified dose descriptor starting point:

NOAEC

Value:

260.089 mg/m³

Explanation for the modification of the dose descriptor starting point:

NOAEL(rat) subacute oral toxicity study: 300 mg/kg bw/day Correction of the starting point according to ECHA Guidance: There are no quantitative data on absorption rates for oral or inhalation route available. Based on the available systemic toxicity following oral application, log Pow in the range of 1.82 - 3.72 and high solubility in water absorption by the respiratory tract is likely and can be assumed to be in the same range than oral absorption. Therefore there is no need to introduce further default values for extrapolation from oral route to inhalation route. Corrected inhalatory NOAEC = NOAEL(oral) x 1/1.15 x 1 Corrected NOAEC (general population) = 260.0869 mg/m³.

AF for dose response relationship:

1

Justification:

The NOAEL is derived from a GLP study according to OFCD TG 407 and was evaluated with Klimisch sore 1. Consequently to this high quality study which complies with the requirements of today ECHA proposes a default assessment factor of 1

AF for differences in duration of exposure:

6

Justification:

Sub-acute to chronic exposure

AF for interspecies differences (allometric scaling):

1

Justification:

Allometric scaling rat versus human: the AF of 4 is already included in the route to route extrapolation

AF for other interspecies differences:

1

Justification:

In an evaluation by ECETOC 2003 and 2010 it is considered that routine application of the factor 2.5 is scientifically unjustified as a default factor

AF for intraspecies differences:

10

Justification:

The NOAEL is not the limit dose and absorption of the test substance can be assumed as demonstrated by log Pow and solubility in water although the molecular weight of Bayscript Blaukomponente cannot be given. Thus, the standard default factor as suggested by ECHA is applied

AF for the quality of the whole database:

1

Justification:

There is information available to cover all relevant toxicological endpoints. The available studies are at most performed according to guideline and GLP.

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

8.7 mg/m³

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

DNEL extrapolated from long term DNEL

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

DNEL related information

General Population - Hazard via dermal route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

1.25 mg/kg bw/day

Most sensitive endpoint:

acute toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

240

Modified dose descriptor starting point:

NOAEL

Value:

300 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

There is no long term dermal study available. Therefore, the oral NOAEL of the sub-acute toxicity study has to be considered instead. According to ECHA Guidance Document, Chapter R8 Version 2.1, December 2012, no additional default factors have to be introduced when performing oral-to-dermal extrapolation because it can be assumed that dermal absorption will not be higher than oral absorption. Thus the NOAEL (oral) = NOAEL(dermal).

AF for dose response relationship:

1

Justification:

The NOAEL is derived from a GLP study according to OECD TG 407 and was evaluated with Klimisch score 1 . Consequently to this high quality study which complies with the requirements of today ECHA proposes a default asssessment factor of 1

AF for differences in duration of exposure:

6

Justification:

Sub-acute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

Allometric scaling rat versus human

AF for other interspecies differences:

1

Justification:

In an evaluation by ECETOC 2003 and 2010 it is considered that routine application of the factor 2.5 as a default factor is scientifically unjustified.

AF for intraspecies differences:

10

Justification:

The NOAEL is not the limit dose and absorption of the test substance can be assumed as demonstrated by log Pow and solubility in water although the molecular weight of Bayscript Blaukomponente cannot be given. Thus the standard default factor as suggested by ECHA is applied.

AF for the quality of the whole database:

1

Justification:

There is information available to cover all relevant toxicological endpoints. The available studies are at most performed according to guideline and GLP.

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

2.5 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

DNEL extrapolated from long term DNEL

Local effects

Long term exposure

Hazard assessment conclusion:

no hazard identified

Acute/short term exposure

Hazard assessment conclusion:

no hazard identified

General Population - Hazard via oral route

Systemic effects

Long term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

1.25 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

Overall assessment factor (AF):

240

Modified dose descriptor starting point:

NOAEL

Value:

300 mg/kg bw/day

Explanation for the modification of the dose descriptor starting point:

No route to route extrapolation necessary.

AF for dose response relationship:

1

Justification:

The NOAEL is derived from a GLP study according to OFCD TG 407 and was evaluated with Klimisch sore 1. Consequently to this high quality study which complies with the requirements of today ECHA proposes a default assessment factor of 1

AF for differences in duration of exposure:

6

Justification:

Sub-acute to chronic exposure

AF for interspecies differences (allometric scaling):

4

Justification:

Allometric scaling rat versus human

AF for other interspecies differences:

1

Justification:

In an evaluation by ECETOC 2003 and 2010 it is considered that routine application of the factor 2.5 as a default factor is scientifically unjustified.

AF for intraspecies differences:

10

Justification:

The NOAEL is not the limit dose and absorption of the test substance can be assumed as demonstrated by log Pow and solubility in water although the molecular weight of Bayscript Blaukomponente cannot be given. Thus the standard default factor as suggested by ECHA is applied.

AF for the quality of the whole database:

1

Justification:

There is information available to cover all relevant toxicological endpoints. The available studies are at most performed according to guideline and GLP.

Acute/short term exposure

Hazard assessment conclusion:

DNEL (Derived No Effect Level)

Value:

2.5 mg/kg bw/day

Most sensitive endpoint:

repeated dose toxicity

Route of original study:

Oral

DNEL related information

DNEL derivation method:

ECHA REACH Guidance

DNEL extrapolated from long term DNEL

General Population - Hazard for the eyes

Local effects

Hazard assessment conclusion:

no hazard identified

Additional information - General Population

Bayscript Blaukomponente

EC-Nr. 916-916-7

DNEL-Derivation: General Population

I. Introduction

Bayscript Blaukomponente is not classified legally.

II. Derivation of DNELs systemic

Basis for DNEL-Derivation

Bayscript Blaukomponente (konserviert) was administered orally by gavage to 5 male and 5 female Wistar rats per dose group using tap water as vehicle, in daily doses of 0, 100, 300, 1000 mg/kg bw/day for a period of four weeks. This GLP study was performed according to OECD guideline 407.

Treatment with Bayscript Blaukomponente resulted in several findings which were related to the color of the test substance. The whole body and several organs of animals treated with 1000 mg/kg bw/day appeared discolored. The kidneys were also discolored in the lower dose groups.

In the kidneys, beneath findings associated with the color of the test substance (starting at 300 mg/kg b.w./day), corticotubular degeneration and increased incidence and/or severity of cortical basophilic (regenerative) tubules were noted at 1000 mg/kg b.w./day. Furthermore, demyelinated fibers in nerve roots at the spinal cord were seen in both sexes at 1000 mg/kg b.w./day.

At 1000 mg/kg b.w./day, histopathological findings in the mesenteric lymph node, the interstitium of the testis, and in the Peyer's patch were related to the colored test item and not accompanied by other structural changes of the organs.

Minor changes in red blood cell parameters observed for males and females at 1000 mg/kg b.w./day were associated with appearing of golden-brown pigment deposition in the spleen in females. In males, the thrombocyte count was increased at 1000 mg/kg b.w./day.

Under the conditions described the no adverse observed effect level (NOAEL) for administration of BAYSCRIPT Blaukomponente (konserviert) to male and female Wistar rats was 300 mg/kg b.w./day mainly due to degenerative effects on kidneys and nerve roots.

1.) General Population DNEL (long term, inhalation route systemic effects)

NOAEL(rat) subacute oral toxicity study: 300 mg/kg bw/day

Correction of the starting point according to ECHA Guidance:

There are no quantitative data on absorption rates for oral or inhalation route available. Based on the available systemic toxicity following oral application, log Pow in the range of 1.82 - 3.72 and high solubility in water absorption by the respiratory tract is likely and can be assumed to be in the same range than oral absorption. Therefore there is no need to introduce further default values for extrapolation from oral route to inhalation route.

Corrected inhalatory NOAEC = NOAEL(oral) x 1/1.15 x 1

Corrected NOAEC (general population) = 260.0869 mg/m³

Overall factor 60

General population DNEL (long term, systemic for inhalation route) 4.35 mg/m³

2.) DNEL short term inhalation route – Systemic effects

There are no data on acute inhalation toxicity available. As acute effects may manifest themselves not immediately but also in considerable time after the exposure ECHA proposes in the Guidance Document R8, 2012, that for DNEL derivation for short term inhalation exposure the long-term DNEL should be used as starting point and a factor of 2 should be applied to extrapolate from long term DNEL to short term DNEL based on available animal data.

Thus, General population DNEL (short term, systemic for inhalation exposure) accounts for 8.7 mg/m³

3.) DNEL (long term, oral route and dermal route – systemic effects)

NOAEL (rat, sub-acute, oral) = 300 mg/kg bw/day

There is no long term dermal study available. Therefore, the oral NOAEL of the sub-acute toxicity study has to be considered instead. According to ECHA Guidance Document, Chapter R8 Version 2.1, December 2012, no additional default factors have to be introduced when performing oral-to-dermal extrapolation because it can be assumed that dermal absorption will not be higher than oral

NOAEL(oral) = NOAEL (dermal) = 300 mg/kg bw/day

Overall factor 240

General population DNEL (long term, systemic for oral route and for dermal route): 1.25 mg/kg bw/day

4.) DNEL (short term, oral route and dermal route) - systemic effects

In the available acute oral rat study all animal tolerated the treatment without mortality and without clinical signs yielding a LD50 value > 2000 mg/kg bw. With respect to dermal application there are acute toxicity studies available using the dermal exposure route. These studies comprise acute toxicity, skin irritation and sensitization. In none of these three study types mortality or signs of intoxication were observed. The LD50 (dermal) is greater 2000 mg/kg bw and the test substance is not irritating to the skin nor caused skin sensitization.

As the NOAEL for the acute oral and for acute dermal toxicity are not determined exactly it is proposed to take the derived DNEL (long term, systemic,oral route and dermal route) as starting point for the derivation of a DNEL (short term, systemic, oral and dermal route). According to ECHA Guidance Document R8 a factor of 2 should be applied to extrapolate from long term DNEL to short term DNEL based on available animal data.

General population- DNEL (short term, systemic for oral route and dermal route) = 2.5 mg/kg bw/day

III. Derivation of DNELs (local)

Basis for derivation DNELs local

The available studies on skin and eye irritation are performed according to OECD TG 404 and 405, respectively, under GLP conditions. Bayscript Blaukomponente was not irritating to the skin of rabbits and was not irritating to the mucous membranes of the eyes of rabbits.

The modified Local Lymph Node Assay (IMDS) was performed according to OECD Guidelines No. 429 and No. 406. The results show that there is no indication for skin sensitizing effect after administration of Bayscript Blaukomponente.

Based on these results no DNELs (local) need to be derived because no hazard was identified

IV. Conclusion (systemic and local)

The relevant DNELs are

Long-term: General population DNEL (systemic inhalation route): 4.35 mg/m³

Long-term: General population DNEL (systemic, oral route and dermal route): 1.25 mg/kg bw/day

Short-term General population DNEL (systemic inhalation route): 8.7 mg/m³

Short-term General population DNEL (systemic oral route and dermal route): 2.5 mg/kg bw/day

No local hazard was identified.