Fatty acids, C18 (even numbered) and C18 unsatd., reaction products with triethanolamine, di-Me sulfate quaternized

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

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

Adequacy of study:

key study

Justification for type of information:

REPORTING FORMAT FOR THE ANALOGUE APPROACH

1. HYPOTHESIS FOR THE ANALOGUE APPROACH
This read-across is based on the hypothesis that source and target substances have similar (eco)toxicological properties because
• they share structural similarities with common functional groups: One quaternised ethanolamine moiety, one to three, mainly two ester groups with a typical UVCB distribution with long-chain fatty acids of natural origin. The molecular structure is almost identical.
• they are manufactured from similar resp. identical precursors (triethanolamine, long-chain fatty acids, dimethyl sulphate) under similar conditions. Therefore common breakdown products via physical and biological processes, which result in structurally similar chemicals are evident
• A constant pattern in the changing of the potency of the properties across the TEA-Esterquats by chain-length and the grade of esterification is not observed, because the fatty acid chain-length distribution is too narrow and similar and the distribution of mono-, di-, and tri-esters is identical. Some variation caused by variation in C=C double bonds may occur and will be discussed at the relevant endpoint.

2. SOURCE AND TARGET CHEMICAL(S) (INCLUDING INFORMATION ON PURITY AND IMPURITIES)
See general justification for read-across attached to chapter 13 of this IUCLID file.

3. ANALOGUE APPROACH JUSTIFICATION
See general justification for read-across attached to chapter 13 of this IUCLID file.

4. DATA MATRIX
See general justification for read-across attached to chapter 13 of this IUCLID file.

Reason / purpose for cross-reference:

read-across: supporting information

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Reason / purpose for cross-reference:

read-across source

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1.91 mg/L

Nominal / measured:

meas. (geom. mean)

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: source substance

Remarks:

partially unsaturated TEA-Esterquat

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

2.1 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Remarks on result:

other: source substance

Remarks:

partially unsaturated TEA-Esterquat

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

11.23 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: absence of respiratory movement and response to physical movement

Remarks on result:

other: source substance

Remarks:

partially unsaturated TEA-Esterquat

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: source substance

Remarks:

partially unsaturated TEA-Esterquat

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

4.9 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Remarks on result:

other: source substance

Remarks:

partially unsaturated TEA-Esterquat

Reference 2

Endpoint:

fish embryo acute toxicity (FET)

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 236 (Fish embryo acute toxicity (FET) test)

Version / remarks:

adopted July 26, 2013

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

Duplicate samples from the freshly prepared test media of the only test concentration and the control were taken at the start of the test.
For the determination of the stability of the test item under the test conditions and of the maintenance of the test item concentrations during the test period, duplicate samples from the test media of the only test concentrations and the control were collected at the end of the test (after 96 hours).
All samples were taken from separate vessels without fish, prepared for analytical samples only.
All samples were stored in a refrigerator (4 ± 4 °C), protected from light until analysis was performed.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
For the pre-conditioning, the only test item concentration of 100 mg test item/L was prepared by dissolving 29.9 mg test item into 299 mL test water by intense stirring for 3 hours and short ultrasonic treatment for 20 minutes. The pre-conditioning media was prepared just before pre-conditioning.
For the main test, the only test item concentration of nominal 100 mg test item/L was prepared by dissolving 50.1 mg test item into 501 mL test water by intense stirring for 2.5 hours and short ultrasonic treatment for 20 minutes.
The test media were prepared just before introduction of the test fish embryos (= start of the test).

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

TEST ORGANISM
- Common name: zebrafish
- Source: In-house breeding
- Age at study initiation (mean and range, SD): Embryos; The cell stage of the embryos introduced at test start ranged from 4 to 8.

FEEDING DURING TEST
- none

The fertilisation rate of the egg batch, determined from a subsample of 72 eggs, was 100 %.

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

214 to 267 mg CaCO3/L

Test temperature:

26.2 to 26.5 °C

pH:

7.8 to 7.9

Dissolved oxygen:

82 to 98 % of the air saturation value

Conductivity:

676 to 725 μS/cm

Nominal and measured concentrations:

100 mg/L analytically verified nominal

Details on test conditions:

TEST SYSTEM
- Test vessel: Polystyrene 24 well plates with 2-5 mL test medium. The well plates were covered with a plastic foil to avoid evaporation of the test media.
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 20
- No. of vessels per control (replicates): 20 (positive control: 4.0 mg/L 3,4-dichloroaniline), 24 (negative control: test water), 4 (internal plate control on each plate: test water)

Eggs were immersed in the well plates containing the only test concentration of the test item, the positive control, the negative control and the internal plate control at cell stage 4 to 8. The eggs were inspected with binoculars in order to determine fertilisation and cell stages. The introduction of the eggs in the 24 well plates indicated the initiation of the test. The plates were incubated for 96 hours.

TEST MEDIUM / WATER PARAMETERS
Reconstituted Water (ISO Medium):
In deionised water (conductivity ≤ 10 μScm-1) analytical grade salts were added to following nominal concentrations:
CaCl2 × 2H2O : 2.0 mmol/L (= 294.0 mg/L)
MgSO4 × 7H2O : 0.5 mmol/L (= 123.0 mg/L)
NaHCO3 : 0.75 mmol/L (= 65.0 mg/L)
KCl : 0.075 mmol/L (= 5.8 mg/L)
Water Hardness : 2.5 mmol/L (= 250.0 mg/L) as CaCO3
Alkalinity : 0.8 mmol/L
Ratio of Ca : Mg = 4 : 1 (based on molarity)
Na : K = 10 : 1 (based on molarity)

OTHER TEST CONDITIONS
- Photoperiod: 16 h light : 8 h dark;
- Light intensity: 560 to 870 lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Apical Observations:
The embryos were observed at test start and approximately after 24, 48, 72 and 96 hours after start of the test for coagulated embryos, lack of somite formation and non-detachment of the tail. The lack of heartbeat was recorded after 48, 72 and 96 hours. All these test parameters were determined using a stereoscopic microscope.

Hatching:
Hatching was recorded in all test chambers after 48, 72, and 96 hours.

TEST CONCENTRATIONS
- Justification for using less concentrations than requested by guideline: Based on teh results of a preliminary study, the main test was carried out as limit test.

Reference substance (positive control):

yes

Remarks:

4.0 mg/L 3,4-dichloroaniline

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

>= 100 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

other: overall effects

Remarks:

one embryo died after 96 hours (most probably not substance related since it is within the mortality range allowed to occur in the control)

Details on results:

After 96 hours, all fish embryos survived in the control. In the only test item concentration of nominal 100 mg test item/L one embryo died after 96 hours. The death of one embryo in the only test item concentration is most probably not substance related since it is within the mortality range allowed to occur in the control (<10%).
Hatching success in the control was 100% and in the only test item concentration 95%. In the only test item concentration of nominal 100 mg test item/L all surviving embryos hatched.
In the internal plate control of nominal 100 mg test item/L all embryos survived and 100% hatched after 96 hours.

Results with reference substance (positive control):

In the positive control, containing 4.0 mg dichloroaniline/L 60% of the embryos died. The remaining eight embryos hatched. In the internal plate control of the positive control, one embryo died, which is still in the acceptable range according to the OECD 236.

Sublethal observations / clinical signs:

Overall Fertilisation Rate: The overall fertilisation rate of all eggs collected was 100 % in the batch tested, thus this validity criterion was met.

Water Temperature: The water temperature was maintained at 26.2 to 26.5°C at the chamber at any time during the test, thus this validity criterion was met.

Survival of Embryos: The overall survival of embryos in the negative (test water) control was 100 % until test termination (96 hours), thus this validity criterion was met.

Hatching Rate: Hatching rate in the negative control was 100 % at test termination (96 hours), thus this validity criterion was met.

Dissolved Oxygen: At test termination (96 hours), the dissolved oxygen concentration in the negative control and the highest test concentration was 82 % of saturation, respectively, thus this validity criterion was met.

Positive Control: The mortality in the positive control was 60 %, thus this validity criterion was met.

Validity criteria fulfilled:

yes

Conclusions:

The 96 h LC50 of C18 and C18 unsatd. TEA-Esterquat to embryonic stages of zebrafish (Danio rerio) was >100 mg a.i./L based on analytically verified nominal concentrations.

Executive summary:

In a 96-h acute toxicity study according to OECD Guideline 236, adopted July 26, 2013, embryonic stages of zebrafish (Danio rerio) were exposed to C18 and C18 unsatd. TEA-Esterquat at nominal concentrations of 0 (control) and 100 mg a.i./L under static conditions.

The test embryos were observed at test start and after approximately 24, 48, 72 and 96 hours test duration for hatching and apical observations (coagulation, somite formation, detachment of tail bud), whereas hatching and heart beat were only observed after 48, 72 and 96 hours.

4 days prior to the test all wells were preconditioned with the test water. Each well was filled with at least 2 mL freshly prepared test water with the respective concentration of the test item, reference item or negative control.

The test concentrations and media used for the preconditioning were discarded before start of the test. All test concentrations and controls were freshly prepared just before introduction of the embryos (= start of the test).

After 96 hours, all fish embryos survived in the control. In the only test item concentration of nominal 100 mg test item/L one embryo died after 96 hours. The death of one embryo in the only test item concentration is most probably not substance related since it is within the mortality range allowed to occur in the control (<10%).

Hatching success in the control was 100% and in the only test item concentration 95%. In the only test item concentration of nominal 100 mg test item/L all surviving embryos hatched.

In the internal plate control of nominal 100 mg test item/L all embryos survived and 100% hatched after 96 hours.

In the positive control, containing 4.0 mg dichloroaniline/L 60% of the embryos died. The remaining eight embryos hatched. In the internal plate control of the positive control, one embryo died, which is still in the acceptable range according to the OECD 236.

The 96-h LC50 was >100 mg a.i./L based on analytically verified nominal concentrations.

 

Results Synopsis

Test Organism Size/Age: Danio rerio embryos, cell stage 4 to 8

Test Type: Static

96 h LC50: >100 mg/L

Endpoint(s) Effected: mortality (coagulation of the embryo, lack of somite formation, the non-detachment of the tail-bud from the yolk sac, lack of heartbeat), hatching 

Description of key information

96 h LC50 > 100 mg a.i/L, based on analytically verified nominal concentration (OECD TG 236, embryonic stages of Danio rerio); GLP; RL1

96 h LC50 = 1.91 mg a.i/L, based on mean measured concentration (OECD TG 203, Oncorhynchus mykiss); GLP; RL1; read-across: partially unsaturated TEA-Esterquat

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1.91 mg/L

Additional information

For the assessment of the acute toxicity to fish, a fish embryo test conducted with the target substance C18 and C18 unsatd. TEA-Esterquat is available. Further reliable relevant data are available for the closely related source substance partially unsaturated TEA-Esterquat. A justification for read-across is given in the Endpoint Summary “Aquatic toxicity”.

 

In a 96-h acute toxicity study according to OECD Guideline 236, adopted July 26, 2013, embryonic stages of zebrafish (Danio rerio) were exposed to C18 and C18 unsatd. TEA-Esterquat at nominal concentrations of 0 (control) and 100 mg a.i./L under static conditions.

The test embryos were observed at test start and after approximately 24, 48, 72 and 96 hours test duration for hatching and apical observations (coagulation, somite formation, detachment of tail bud), whereas hatching and heart beat were only observed after 48, 72 and 96 hours.

4 days prior to the test all wells were preconditioned with the test water. Each well was filled with at least 2 mL freshly prepared test water with the respective concentration of the test item, reference item or negative control.

The test concentrations and media used for the preconditioning were discarded before start of the test. All test concentrations and controls were freshly prepared just before introduction of the embryos (= start of the test).

After 96 hours, all fish embryos survived in the control. In the only test item concentration of nominal 100 mg test item/L one embryo died after 96 hours. The death of one embryo in the only test item concentration is most probably not substance related since it is within the mortality range allowed to occur in the control (<10%).

Hatching success in the control was 100% and in the only test item concentration 95%. In the only test item concentration of nominal 100 mg test item/L all surviving embryos hatched.

In the internal plate control of nominal 100 mg test item/L all embryos survived and 100% hatched after 96 hours.

In the positive control, containing 4.0 mg dichloroaniline/L 60% of the embryos died. The remaining eight embryos hatched. In the internal plate control of the positive control, one embryo died, which is still in the acceptable range according to the OECD 236. The 96-h LC50 was >100 mg a.i./L based on analytically verified nominal concentrations.

 

In a 96-h acute toxicity study conducted according to OECD Guideline 203 (Fish, Acute Toxicity Test), rainbow trouts (Oncorhynchus mykiss) were exposed to partially unsaturated TEA-Esterquat (CAS No. 157905-74-3) at nominal concentrations of 0 (control), 1.0, 1.6, 2.56, 4.1, 6.55, 10.55 mg/L under semi-static conditions. The mean measured concentration was determined to be 59 % of nominal concentrations. The 96-h LC50 was 1.91 mg/L (based on mean measured concentration). Sublethal effects as difficulties with maintainance of balance and unusual behaviour (reduced activity and or orientation to bottom or surface of the test vessels) were observed within the first three hours in the highest test concentration  

 

In a further study performed according to OECD Guideline 203 (Fish, Acute Toxicity Test) under semi-static conditions the LC50 (96 h) of partially unsaturated TEA-Esterquat was determined to be 2.1 mg/L for Danio rerio. The effect concentration (96h-LC50 = 2.6 mg/L) was corrected because the analytical recovery (HPLC) of test substance was below 80% of nominal concentration.

Partially unsaturated TEA-Esterquat was classified as toxic to Danio rerio by the study authors. However, according to CLP (EU-GHS), Regulation (EC) No 1272/2008 the criteria for acute aquatic toxicity of fish (96h LC50) is </= 1 mg/L.

 

There are two further reliable studies available for partially unsaturated TEA-Esterquat performed according to OECD Guideline 203 (Fish, Acute Toxicity Test) under semi-static conditions and a study according to OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study) under flow-through conditions, but all performed without analytical control. The LC50 (96 h) effect concentrations were 7.48 mg/L (test mat., nominal, based on a.i. > 80 % the 96h LC50 was > 5.98 mg a.i./L) for Danio rerio and 11.23 mg a.i./L and 14.45 mg a.i./L for Oncorhynchus mykiss each, respectively. The nominal effect concentrations were corrected for the content of active ingredient.

One study is available for Leuciscus idus (Golden orfe). In this study the 24 to 96 h LC 50 of TEA-Esterquat partially unsaturated was 5.1 mg test mat./L (nominal).

In a flow through test according to OECD Guideline 204 (Fish, Prolonged Toxicity Test: 14-day Study, 1984) the 96 h and 14 d LC50 of partially unsaturated TEA-Esterquat were 4.9 mg/L each (nominal) for Danio rerio.

 

The lowest 96 h LC50 in fish of 1.91 mg/L obtained in rainbow trouts (Oncorhynchus mykiss) will be used for chemical safety assessment.