2-Octen-1-ylsuccinic anhydride, mixture of cis and trans

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

2013-03-19 to 2013-03-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Justification for type of information:

A category approach will be used for the hazard assessment of several endpoints. The hypothesis for the category of C8-12 Alkenyl Succinic Anhydrides is that data can be read-across among members of the category because their properties and behaviours are similar, based on common functional groups and similar breakdown products, and based on a constant pattern in changing of the potency of properties of the various carbon chain lengths. These features are in accordance with Annex XI, Section 1.5, of Regulation EC No. 1907/2006.  Common functional groups are: a dihydro-2,5-Furandione (cyclic anhydride) ring, carbon chain of length 8 to 12 carbons, with or without branching alkyl groups, and a single carbon-carbon double bond in the carbon chain, location unspecified. There are no additional functional groups which would contribute incremental or different toxicity.
 
For each category member, the breakdown product is the dioic acid of the corresponding anhydride; these dioic acids also have common functional groups. A constant pattern may also be displayed in acute toxicity, dermal irritancy and biodegradation, with the lowest carbon chain length (C8) displaying the highest activity. Irritation, toxicity and degradation potential diminish with increasing carbon chain length. Read-across among the category members is substantiated by the common behaviour in physico-chemical and toxicity behaviours, as provided in the Chemical Category Report Format (CCRF) attached to the IUCLID file. It is adequate to fulfil the information requirements of Annex IX, to be the basis for classification and labelling decisions, and for risk assessment.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations:Limit Test with 100 mg/L
- Sampling method:
The limit concentration and the control were analytically verified by LC-MS/MS at the start (0 h) and at the end of the exposure (48 h) as specified below. At the start of the exposure (0 h), samples were taken immediately after preparation of the limit concentration and analysed. At the end of the exposure (48 h), samples were taken directly from the test vessels, which contained the daphnids.The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared in demineralised water one day prior to the start of the exposure. 100 mg/L of the test item were weighed out and transferred with an appropriate volume of demineralised water into a measuring flask. The measuring flask was not completely filled up with demineralised water, because the mineral salts of the dilution water had to be added immediately before the start of the exposure phase. For the transfer of the weighed test item into the measuring flask, ultrasound was used for approximately 1 minute. The stock solution was stirred with approximately 1100 rpm for 24 hours at room temperature.
After the stirring period, the mineral components as specified under 'Dilution water' were added to yield the limit test concentration.


- Eluate: Dilution water
- Differential loading: 100 mg/L
- Controls: Dilution water without test item tested under the same conditions as the test goup.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
The succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) was expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. ELENDT, 1990), was set to a total hardness of approximately 140 mg CaCO3/L still acceptable to achieve valid results.

Component Concentration [mg/L]
CaCl2 x 2 H2O 147
MgSO4 x 7 H2O 123
KCl 5.80
NaHCO3 64.8
Na2SiO3 4.30
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18
Na2EDTA x 2 H2O 5.00
FeSO4 x 7 H2O 1.99
H3BO3 0.29
MnCl2 x 4 H2O 0.36
LiCl 0.30
SrCl2 x 6 H2O 0.15
RbCl 0.071
NaBr 0.016
Na2MoO4 x 2 H2O 0.063
CuCl x 2 H2O 0.017
ZnCl2 0.013
CoCl2 x 6 H2O 0.010
KJ 0.00325
Na2SeO3 0.00219
NH4VO3 0.000575
Thiaminhydrochloride 0.075
Cyanocobalamine 0.0010
Biotin 0.00075
pH 8.2  0.8

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 144 (mg CaCO3/L)

Test temperature:

19.8 °C (Water)

pH:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal test item
concentration
[mg/L] pH-Value

100 6.39
Control 8.32

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal test item
concentration

[mg/L] pH-Values
Replicates
1 2 3 4
100 7.53 7.39 7.29 7.18
Control 7.70 7.70 7.69 7.68

Dissolved oxygen:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal test item
concentration
[mg/L] Dissolved O2-Concentration [mg/L]
100 9.01
Control 8.99

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal test item
concentration
[mg/L] Dissolved O2 -Concentration [mg/L]
Replicates
1 2 3 4
100 7.81 8.03 8.13 7.74
Control 6.56 7.54 7.60 7.45

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

please refer to "Any other information on materials and methods"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start:421 µS/cm
- Culture medium different from test medium: he succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) was expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. ELENDT, 1990), was set to a total hardness of approximately 140 mg CaCO3/L still acceptable to achieve valid results.
- Intervals of water quality measurement: At test start and test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilization Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal
Test Item
Concentration
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: Not applicable

Details on results:

- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.60 (CI 1.54 - 1.65) mg/L

Reported statistics and error estimates:

The effects of the limit concentration were deduced directly from the observed immobilisation rates.
An EC50-value was only calculated for the reference item by sigmoidal dose-response regression. The confidence interval for EC50 will be calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism5.

Immobilisation Rates of the Definitive Test after 24 and 48 h of Exposure

(n = 20, divided into 4 replicates with 5 daphnids each)

Nominal test item concentration [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

At the limit concentration of 100 mg/L of the test item Tripropenyl succinic acid, no effects were observed on Daphnia magna. Data can be read-across among members of the C8-12 Alkenyl Succinic Anhydrides Category, based on common functional groups, similar break-down products and potency patterns among carbon-chain length. This is adequate to fulfil the information requirements, to be the basis for classification and labelling decisions, and for risk assessment.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the limit concentration of 100 mg/L of the test item Tripropenyl succinic acid(batch number: 12-SK002) were determined according to OECD 202 (2004). The limit test was conducted under static conditionsover a period of 48 hours from 2013-03-19 to 2013-03-22, with the definitive exposure phase from 2013-03-20 to 2013-03-22, at Dr. U. Noack-Laboratorien, Käthe-Paulus-Str. 1,31157 Sarstedt, Germany .

The succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) is expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. Elendt, 1990), was set to a total hardness of approximately 140 mg CaCO₃/L still acceptable to achieve valid results. Twenty daphnids were exposed to the limit concentration and the control.

The test item Tripropenyl succinic acidwas analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) at the limit concentration of 100 mg/L and the control.

The measured concentration of the test item at the start of the exposure (0 h) was 100 % of the nominal value of the limit concentration. The measured concentration at the end of the exposure (48 h) was 99 % of the nominal value of the limit concentration. This demonstrates that the test item concentration was successfully maintained for the duration of the test.

The effect levels are based on the nominal concentration of 100 mg/L of the test item Tripropenyl succinic acid.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guidelines were fulfilled.

EC-Values, NOEC and LOEC

(based on the nominal concentrations of the test item Tripropenyl succinic acid)

Tripropenyl succinic acid
	Test duration [h]	Nominal test item concentration [mg/L]	Confidence interval (p = 95%) [mg/L]
EC10 / EC50 / EC100	24	> 100	Not applicable
	48	> 100	Not applicable
NOEC	48	100	---
LOEC	48	> 100	---

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2013-03-19 to 2013-03-22

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

yes

Details on sampling:

- Concentrations:Limit Test with 100 mg/L
- Sampling method:
The limit concentration and the control were analytically verified by LC-MS/MS at the start (0 h) and at the end of the exposure (48 h) as specified below. At the start of the exposure (0 h), samples were taken immediately after preparation of the limit concentration and analysed. At the end of the exposure (48 h), samples were taken directly from the test vessels, which contained the daphnids.The method was validated prior to this study according to SANCO 3029/99 rev.4 (2000-07-11).
- Sample storage conditions before analysis: All original samples were stored at room temperature until sample preparation. Prepared samples were stored at room temperature until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: A stock solution was prepared in demineralised water one day prior to the start of the exposure. 100 mg/L of the test item were weighed out and transferred with an appropriate volume of demineralised water into a measuring flask. The measuring flask was not completely filled up with demineralised water, because the mineral salts of the dilution water had to be added immediately before the start of the exposure phase. For the transfer of the weighed test item into the measuring flask, ultrasound was used for approximately 1 minute. The stock solution was stirred with approximately 1100 rpm for 24 hours at room temperature.
After the stirring period, the mineral components as specified under 'Dilution water' were added to yield the limit test concentration.


- Eluate: Dilution water
- Differential loading: 100 mg/L
- Controls: Dilution water without test item tested under the same conditions as the test goup.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna (STRAUS)
- Strain: Clone 5
- Source: Institut für Wasser-, Boden- Lufthygiene (WaBoLu), further own breeding
- Age at study initiation (mean and range, SD): 2 to 24 h
- Method of breeding: In 2-3 L glass vessels with appr. 1.8 L culture medium, at 20 +/- 2 °C, in an incubator, 16 h illumination, illumination strength max. 20 µExm-2 x s-1
- Feeding during test: No feeding

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): Same as test
- Type and amount of food: No feeding
- Health during acclimation (any mortality observed): Healthy, no mortality observed

DILUTION WATER:
The succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) was expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. ELENDT, 1990), was set to a total hardness of approximately 140 mg CaCO3/L still acceptable to achieve valid results.

Component Concentration [mg/L]
CaCl2 x 2 H2O 147
MgSO4 x 7 H2O 123
KCl 5.80
NaHCO3 64.8
Na2SiO3 4.30
NaNO3 0.27
KH2PO4 0.14
K2HPO4 0.18
Na2EDTA x 2 H2O 5.00
FeSO4 x 7 H2O 1.99
H3BO3 0.29
MnCl2 x 4 H2O 0.36
LiCl 0.30
SrCl2 x 6 H2O 0.15
RbCl 0.071
NaBr 0.016
Na2MoO4 x 2 H2O 0.063
CuCl x 2 H2O 0.017
ZnCl2 0.013
CoCl2 x 6 H2O 0.010
KJ 0.00325
Na2SeO3 0.00219
NH4VO3 0.000575
Thiaminhydrochloride 0.075
Cyanocobalamine 0.0010
Biotin 0.00075
pH 8.2  0.8

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Post exposure observation period:

No

Hardness:

Dilution water at test start: 144 (mg CaCO3/L)

Test temperature:

19.8 °C (Water)

pH:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal test item
concentration
[mg/L] pH-Value

100 6.39
Control 8.32

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal test item
concentration

[mg/L] pH-Values
Replicates
1 2 3 4
100 7.53 7.39 7.29 7.18
Control 7.70 7.70 7.69 7.68

Dissolved oxygen:

Water Quality Parameters at the Start of the Exposure (0 h)
(measured in one additional replicate per concentration level and control)
Nominal test item
concentration
[mg/L] Dissolved O2-Concentration [mg/L]
100 9.01
Control 8.99

Water Quality Parameters at the End of the Exposure (48 h)
(measured in all replicates)
Nominal test item
concentration
[mg/L] Dissolved O2 -Concentration [mg/L]
Replicates
1 2 3 4
100 7.81 8.03 8.13 7.74
Control 6.56 7.54 7.60 7.45

Salinity:

Not measured, freshwater

Nominal and measured concentrations:

please refer to "Any other information on materials and methods"

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers, 50 mL
- Type (delete if not applicable): closed with parafilm
- Material, size, headspace, fill volume: Glass beakers (4 cm ID x 7 cm H), 50 mL
- Aeration: No
- Renewal rate of test solution (frequency/flow rate): static test
- No. of organisms per vessel: 5
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4
- No. of vessels per vehicle control (replicates): None


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: see above
- Conductivity: Dilution water at test start:421 µS/cm
- Culture medium different from test medium: he succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) was expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. ELENDT, 1990), was set to a total hardness of approximately 140 mg CaCO3/L still acceptable to achieve valid results.
- Intervals of water quality measurement: At test start and test end


OTHER TEST CONDITIONS
- Adjustment of pH: No
- Photoperiod: 16:8 light:dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE x m-2 x s-1


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Immobility was determined in all groups after 24 h and 48 h.

TEST CONCENTRATIONS
- Range finding study:
Immobilization Rates in the non GLP Preliminary Range Finding Test
(n = 20, divided into 2 replicates with 10 daphnids each)
Nominal
Test Item
Concentration
[mg/L] IMMOBILISATION [%]
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
100 0 0 0 0 0 0
Control 0 0 0 0 0 0

Reference substance (positive control):

yes

Remarks:

Potassium dichromate

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mobility

Remarks on result:

other: Not applicable

Details on results:

- Mortality of control: 0 %
- Any observations (e.g. precipitation) that might cause a difference between measured and nominal values: No

Results with reference substance (positive control):

- Results with reference substance valid? Yes
- EC50/LC50: 1.60 (CI 1.54 - 1.65) mg/L

Reported statistics and error estimates:

The effects of the limit concentration were deduced directly from the observed immobilisation rates.
An EC50-value was only calculated for the reference item by sigmoidal dose-response regression. The confidence interval for EC50 will be calculated from the best-fit values, the standard error and the t-distribution with the software GraphPad prism5.

Immobilisation Rates of the Definitive Test after 24 and 48 h of Exposure

(n = 20, divided into 4 replicates with 5 daphnids each)

Nominal test item concentration [mg/L]	IMMOBILISATION [%]
	24 h					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

Validity criteria fulfilled:

yes

Conclusions:

At the limit concentration of 100 mg/L of the test item Tripropenyl succinic acid, no effects were observed on Daphnia magna. Data can be read-across among members of the C8-12 Alkenyl Succinic Anhydrides Category, based on common functional groups, similar break-down products and potency patterns among carbon-chain length. This is adequate to fulfil the information requirements, to be the basis for classification and labelling decisions, and for risk assessment.

Executive summary:

In the acute immobilisation test with Daphnia magna (STRAUS), the effects of the limit concentration of 100 mg/L of the test item Tripropenyl succinic acid(batch number: 12-SK002) were determined according to OECD 202 (2004). The limit test was conducted under static conditionsover a period of 48 hours from 2013-03-19 to 2013-03-22, with the definitive exposure phase from 2013-03-20 to 2013-03-22, at Dr. U. Noack-Laboratorien, Käthe-Paulus-Str. 1,31157 Sarstedt, Germany .

The succinic acid derivate typically forms a calcium salt with very low solubility (ca. 40 ppm) removing most of the test item from the aqueous solution by precipitation. Therefore, the recovery of the test item at the end of an exposure interval (48 h) is expected to be very low and the guideline quality criteria for the analytical recovery are no longer fulfilled. To reduce the precipitation of Calcium salts, the total hardness in the Elendt M4 medium (B.P. Elendt, 1990), was set to a total hardness of approximately 140 mg CaCO₃/L still acceptable to achieve valid results. Twenty daphnids were exposed to the limit concentration and the control.

The test item Tripropenyl succinic acidwas analytically verified by LC-MS/MS at the start of the exposure (0 h) and at the end of the exposure (48 h) at the limit concentration of 100 mg/L and the control.

The measured concentration of the test item at the start of the exposure (0 h) was 100 % of the nominal value of the limit concentration. The measured concentration at the end of the exposure (48 h) was 99 % of the nominal value of the limit concentration. This demonstrates that the test item concentration was successfully maintained for the duration of the test.

The effect levels are based on the nominal concentration of 100 mg/L of the test item Tripropenyl succinic acid.

Water quality parameters (i.e. pH-value and dissolved oxygen concentration), measured at the start (0 h) and at the end of the exposure (48 h), were within the acceptable limits. The validity criteria of the test guidelines were fulfilled.

EC-Values, NOEC and LOEC

(based on the nominal concentrations of the test item Tripropenyl succinic acid)

Tripropenyl succinic acid
	Test duration [h]	Nominal test item concentration [mg/L]	Confidence interval (p = 95%) [mg/L]
EC10 / EC50 / EC100	24	> 100	Not applicable
	48	> 100	Not applicable
NOEC	48	100	---
LOEC	48	> 100	---

Description of key information

An acute daphnid toxicity study was conducted in accordance with OECD Guideline 202 on tripropenyl succinic acid (CAS RN 147384-84-7), by read-across based on a grouping of substances (category approach). 

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

100 mg/L

Additional information

An acute daphnid toxicity study was conducted in accordance with OECD Guideline 202 on tripropenyl succinic acid (CAS RN 147384-84-7), by read-across based on a grouping of substances (category approach). The test was conducted under static conditions as a limit test at 100 mg/L. The NOEC was reported at 100 mg/L, and the EC₅₀was reported as ˃ 100 mg/L.

A category approach is used for the hazard assessment of several endpoints. The hypothesis for the category of C8-12 Alkenyl Succinic Anhydrides is that data can be read-across among members of the category, because the properties and behaviours of category members are similar, based on common functional groups, similar breakdown products, and demonstration of a constant pattern associating the potency of properties with the various carbon chain lengths. Functional groups include a dihydro-2,5-furandione cyclic anhydride ring, a carbon chain of length 8 -12 carbons, and a single carbon-carbon double-bond within the carbon chain. The primary functional group associated with toxicity is the succinic anhydride moiety, which is quickly hydrolysed to form a butanedioic acid. A constant pattern may also be displayed in acute toxicity, dermal irritancy and biodegradation, with the lowest carbon chain length (C8) displaying the highest activity. Irritation, toxicity and degradation potential diminish with increasing carbon chain length. Read-across among the category members is substantiated by the common behaviour in physico-chemical and toxicity behaviours, as provided in the Chemical Category Report Format (CCRF) attached to the IUCLID file. It is adequate to fulfil the information requirements of Annex IX, to be the basis for classification and labelling decisions, and for risk assessment.