1,5-Pentanediamine, 2-methyl-, reaction products with 2-ethyl-1,4-butanediamine and glycidyl tolyl ether

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

20 May 2018 - 20 June 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Version / remarks:

1992

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: freshly prepared test preparations at 0 and 72 hours, old or expired test preparations at 24 and 96 hours
- Sample storage conditions before analysis: The samples were stored frozen prior to analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
Due to the low aqueous solubility and complex nature of the test item, for the purposes of the study the test medium was prepared as a WAF of the test item.
A stirring period of 23 hours followed by a 1 Hour standing period was employed to ensure an equilibrated concentration of dissolved test item in the aqueous phase.

In accordance with the recommendations of REACh, the test was conducted according to the threshold approach recommended by ECHA. Using this approach the lowest EL50 value from either the Algal Growth Inhibition study or Acute Toxicity to Daphnia magna study is set as the threshold loading rate and a “Limit test” is conducted at this threshold loading rate. If no mortalities are observed this indicates that fish are not the most sensitive species and that the LL50 is greater than the threshold loading rate. The EL50 value obtained from the Algal Growth Inhibition study (Envigo study number FF65QT) was the lowest of these two EL50 values and hence the test was conducted at a single loading rate of 0.11 mg/L.

A nominal amount of test item (22 mg) was added to the surface of 22 liters of test water to give the 1.0 mg/L loading rate. After the addition of the test item, the test water was stirred by magnetic stirrer using a stirring rate such that a vortex was formed to give a dimple at the water surface. The stirring was stopped after 23 hours and the mixture allowed to stand for 1-Hour. A wide bore glass tube, covered at one end with Nescofilm was submerged into the vessel, sealed end down, to a depth of approximately 5 cm from the bottom of the vessel. A length of PVC tubing was inserted into the glass tube and pushed through the Nescofilm seal. Microscopic inspection of the WAF showed no micro dispersions or undissolved test item to be present. The aqueous phase or WAF was removed by mid depth siphoning (the first 75 to 100 mL discarded) to give the 1.0 mg/L loading rate WAF. This solution was then diluted further to give the 0.11 mg/L loading rate.
The test preparation was mixed with a flat bladed stirrer for approximately 1 minute to ensure adequate mixing and homogeneity.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK; maintained in house since 12 April 2018
- fish had a mean standard length of 4.5 cm (standard deviation = 0.18) and a mean weight of 0.85 g (standard deviation = 0.10) at the end of the definitive test
- Maintenance of the brood fish: glass fiber tank with a "single pass" water renewal system

ACCLIMATION
- Acclimation period: 14 May 2018 to 21 May 2018
- Acclimation conditions (same as test or not): same
- Type and amount of food during acclimation: stock fish were fed commercial trout pellets, which was discontinued approximately 25 hours prior to the start of the definitive test
- Health during acclimation (any mortality observed): no mortality

FEEDING DURING TEST
no

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

approx. 140 mg/L as CaCO3

Test temperature:

14°C to 15°C

pH:

7.7-8.1

Dissolved oxygen:

>/= 9.0 mg O2/L

Nominal and measured concentrations:

nominal 0, 0.11 mg/L (loading rate WAF)

Details on test conditions:

TEST SYSTEM
- Test vessel: 25 to 30 liter glass exposure vessels containing 20 liters of test media
- Type (delete if not applicable): closed
- Aeration: test vessels were aerated via narrow bore glass tubes
- Renewal rate of test solution: daily renewal
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1
- Biomass loading rate: 0.30 g body weight/liter

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Laboratory tap water was dechlorinated by passage through an activated carbon filter (Purite Series 500) and partly softened (Elga Nimbus 1248D Duplex Water Softener) giving water with a total hardness of approximately 140 mg/L as CaCO3.


OTHER TEST CONDITIONS
- Adjustment of pH:
- Photoperiod: 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods
- Light intensity:

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :

TEST CONCENTRATIONS
- Spacing factor for test concentrations:
- Justification for using less concentrations than requested by guideline:
- Range finding study
- Test concentrations:
- Results used to determine the conditions for the definitive study:

Reference substance (positive control):

no

Duration:

96 h

Dose descriptor:

EL50

Effect conc.:

> 0.11 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

loading rate WAF

Basis for effect:

mortality (fish)

Details on results:

- There were no sub lethal effects of exposure observed.
- Mortality of control: no mortality in the control
- Effect concentrations exceeding solubility of substance in test medium: At the start and throughout the test all control solutions were observed to be clear colorless solutions.

Results with reference substance (positive control):

n.a.

Reported statistics and error estimates:

n.a.

Sublethal observations / clinical signs:

Cumulative Mortality Data in the Definitive Test

Nominal Loading Rate (mg/L)	Cumulative Mortality (Initial Population = 7)							Mortality (%)
	1 Hour	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	96 Hours	96 Hours
Control	0	0	0	0	0	0	0	0
0.11	0	0	0	0	0	0	0	0

 

Sub-lethal Effects of Exposure in the Definitive Test

Nominal Loading Rate (mg/L)	Sub-lethal Effects	Time (Hours)
		1	3	6	24	48	72	96
Control	No abnormalities detected	7	7	7	7	7	7	7
0.11	No abnormalities detected	7	7	7	7	7	7	7

 

 

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of CGE-P;DA adduct to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated using the threshold approach and gave a 96 Hour LL50 value of greater than 0.11 mg/L loading rate WAF. The NOEL rate was 0.11 mg/L loading rate WAF.

Executive summary:

This study was performed to assess the acute toxicity of CGE-PMDA adduct to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with OECD Guideline 203 (1992) and EU method C.1 (2008).

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

In accordance with the recommendations of REACh, the test was conducted according to the threshold approach recommended by ECHA. Using this approach, the lowest EL50 value from either the Algal Growth Inhibition study or Acute Toxicity to Daphnia magna study is set as the threshold loading rate and a “Limit test” is conducted at this threshold loading rate. If no mortalities are observed this indicates that fish are not the most sensitive species and that the LL50 is greater than the threshold loading rate. The EL50 value obtained from the Algal Growth Inhibition study (Envigo study number FF65QT) was the lowest of these two EL50 values and hence the test was conducted at a single loading rate of 0.11 mg/L.

Seven fish were exposed to a WAF of the test item, at a single nominal loading rate of0.11 mg/L for a period of 96 hours at a temperature of 14 °C to 15 ºC under semi‑static test conditions. The number of mortalities and any sub‑lethal effects of exposure in each test and control vessel were determined 1, 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Chemical analysis of the freshly prepared test preparations at 0 and 72 hours showed measured test concentrations of 0.12 and 0.061 mg/L respectively. Chemical analysis of the old or expired test preparations at 24 and 96 hours showed measured test concentrations of 0.098 and 0.053 mg/L respectively.

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

Exposure of rainbow trout to the test item gave LL50 values of greater than 0.11 mg/L loading rate WAF. The No Observed Effect Loading Rate (NOEL) was 0.11 mg/L loading rate WAF.

Description of key information

96 h LL50 > 0.11 mg/L (OECD TG 203; Oncorhynchus mykiss; WAF, limit test)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

0.11 mg/L

Additional information

This study was performed to assess the acute toxicity of CGE-PMDA adduct to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with OECD Guideline 203 (1992) and EU method C.1 (2008).

Due to the low aqueous solubility and complex nature of the test item, for the purposes of the test, the test medium was prepared as a Water Accommodated Fraction (WAF).

In accordance with the recommendations of REACh, the test was conducted according to the threshold approach recommended by ECHA. Using this approach, the lowest EL50 value from either the Algal Growth Inhibition study or Acute Toxicity to Daphnia magna study is set as the threshold loading rate and a “Limit test” is conducted at this threshold loading rate. If no mortalities are observed this indicates that fish are not the most sensitive species and that the LL50 is greater than the threshold loading rate. The EL50 value obtained from the Algal Growth Inhibition study (Envigo study number FF65QT) was the lowest of these two EL50 values and hence the test was conducted at a single loading rate of 0.11 mg/L.

Seven fish were exposed to a WAF of the test item, at a single nominal loading rate of0.11 mg/L for a period of 96 hours at a temperature of 14 °C to 15 ºC under semi‑static test conditions. The number of mortalities and any sub‑lethal effects of exposure in each test and control vessel were determined 1, 3 and 6 hours after the start of exposure and then daily throughout the test until termination after 96 hours.

Chemical analysis of the freshly prepared test preparations at 0 and 72 hours showed measured test concentrations of 0.12 and 0.061 mg/L respectively. Chemical analysis of the old or expired test preparations at 24 and 96 hours showed measured test concentrations of 0.098 and 0.053 mg/L respectively.

Given that the toxicity cannot be attributed to a single component or a mixture of components, but to the test item as a whole, the results were based on nominal loading rates only.

Exposure of rainbow trout to the test item gave LL50 values of greater than 0.11 mg/L loading rate WAF. The No Observed Effect Loading Rate (NOEL) was 0.11 mg/L loading rate WAF.