Sodium hydrogen N-(1-oxotetradecyl)-L-glutamate

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

From 07 March, 2016 to 16 May, 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Version / remarks:

2012

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Remarks:

Quality Assesment is on page 4 of the attached report

Specific details on test material used for the study:

Batch no.: 3837 supplied by the sponsor
25.5% in water as active ingredient (water solution), purity 100%
Solubility: soluble in water
Storage condition of test material: room temperature (15 °C - 25 °C)
Stability and homogeneity of the test material in the vehicle/solvent under test conditions (e.g. in the exposure medium) and during storage: stable for 12 months minimum
Solubility: soluble and stable in water
Reactivity of the test material with the incubation material used (e.g. plastic ware): not reactive
pH (20 °C) = 6.22
More information can be found on the attached report

Analytical monitoring:

yes

Remarks:

Determination of analytical concentration of total organic carbon (TOC), see details on sampling for more information

Details on sampling:

Samples, properly diluted with deionised water with low TOC contents, have been analysed by means of an integrated sampling system directly from vials containing 40 ml volume. For each sample under test the corresponding blank is analysed.
CALCULATIONS
TOCsub (t=0) = TOCsub t = 0 -TOC (blank t=0 -dilution water t=0)
TOCsub (t) = TOCsub t -TOC (blank t -dilution water t)
% stability: 100 x (TOC test subst. t - TOC test subst t=0)
Samples are considered stable if the % stability is between 80 and 120% of the value.
STABILITY
The samples were stable throughout the test period. Results that are summarized in table 5 of the attached report confirm stability.
Preliminaty stability 82%
Stability 1-2 week 113%
Stability 2-3 week 116%

Vehicle:

yes

Remarks:

Substance is alreday in water solution and dilution with specific salts are prepared

Details on test solutions:

As dilution water, reconstituted water has been used. It has been prepared dissolving salts in 1 l of deionised water as described by OECD 211 and as reported on page 8 of the attached report.
Separate stock solutions of individual traces elements are first prepared in water of suitable purity, e.g. deionised, distilled or reverse osmosis. From these different stock solutions a second single stock solution is prepared, which contains all trace elements (combined solutions). From these solutions, M4 or M7 solutions are prepared adding also macronutrients and vitamins. Vitamins have been added to the media shortly before use. To avoid salts precipitations, the aliquots of stock solutions have been added to about 500-800 ml f deionised water and then fill it up to 1 liter.
For the study as dilution water medium M4 has been used. Details are on page 8 f the attached report.

Test organisms (species):

Daphnia magna

Details on test organisms:

Age: < 24 hours
Origine: Breeder in Eurofins Biolab Test Facility
Feeding: green alga
Number of animals: 50
Water change: 3 times per week
Temperature: 20 +/-2 °C
Lightning: 16 hours of light and 8 of darkness
Water hardness: < 250 mg CaCO3/l

Test type:

semi-static

Water media type:

other: deionized water + dissolved salts and nutrients

Limit test:

yes

Total exposure duration:

21 d

Hardness:

> 140 mg CaCO3/l

Test temperature:

20 °C +/- 2 °C

pH:

6.0 - 9.0
pH must not change of more than 1.5 units during the test

Dissolved oxygen:

> 3 mg/l
Areation is absent during the test

Nominal and measured concentrations:

Different concentrations have been used for range finding test, 1 for definitive limit test (10 mg/l active concentration). 1 g/l of active substance is 3.920 g/l of the tests item. For range finding 3.9209 g of the test item has been waited in order to obtain stock solution A that has been diluted to 200 mg/l of active ingredient (B).5 different concentrations of active ingredient has been prepared with ten-fold dilutions form 100 mg/l to 0.01 mg/l. As after the range finding no effects were observed at 10 mg/l, the definitive test limit has been performed at this concentration.

Details on test conditions:

For range finding 30 Daphnie have been used, 25 treated and 5 used as control, kept in same conditions. For definitive test 10 Daphnie were used and 10 used as control. Every 24 hours from the beginning immobilization and other effects were evaluated. The offspring have been counted and removed daily. Number of aborted eggs and dead offspring have been recorded. Also time to production of first brood has been assessed. Immobilization of Daphnie has been evaluated verifying if the organism after light stirring of the vessel won't be capable to move in the following 15 seconds. Every immoblized organisms have been removed from the vessel. At the beginning and at least 3 times/week dissolved O2, pH and temperature were measured and recorded.

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

LOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

immobilisation

Key result

Duration:

21 d

Dose descriptor:

NOEC

Effect conc.:

> 10 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

immobilisation

Details on results:

Quality criteria have been satisfied.
The immobilization of the parent animals in control doesn't exceed 20% at the end of the test.
The mean number of live offspring produced in control per parent animal surviving at the end of the test is >=60.
The coefficient of variation around the mean of living offsrping produced per parent animal in control doesn't exceed 25%.
Mean number of live offspring produced per parent animal in control surviving at the end of the test: 248
Coefficient of variation in control: 11.4%
Time to production of first brood in control: 9 days.
Oxygen, pH and temperature were all in validity test range (tables 2, 3 and 4 of the attached report). TOC at the beginning and at the end of the test was in the validity criteria range (table 5).
See results tables reported in any other information

Reported statistics and error estimates:

Statistical evaluation (see Attachment 2 of the attached report) was done using Pharm/PCS Version 4.2.

Table 1 number of immobilised Daphnie and total number of offspring produced by every organism

No immobilized parent Daphnie in treated replica

CONTROL -OFFSPRING PRODUCED

Control	Total number offspring	Immobilization day
1	231	-
2	285	-
3	253	-
4	265	-
5	217	-
6	216	-
7	231	-
8	295	-
9	264	-
10	225	-

TREATED (10 MG/L) OFFSPRING PRODUCED

 

TREATED  REPLICA	NUMBER OFFSPRING	IMMOBILIZATION DAY
1	278	-
2	254	-
3	256	-
4	194	-
5	184	-
6	269	-
7	282	-
8	233	-
9	195	-
10	257	-

Validity criteria fulfilled:

yes

Conclusions:

Under the study conditions (OECD 211) the substance has a LOEC and NOEC > 10 mg/l after 21 days.

Executive summary:

A study was conducted to determine the effect on reproduction of the substance to Daphnia magna according to OECD Guideline 211, in compliance with GLP. Daphnia magna were exposed to a series of concentrations of the test substance for 21 days to find the limit concentration. Total organic carbon (TOC) concentrations were measured with a TOC analyser and the test substance was considered stable over the duration of the test. Daphnie have been kept under observation for a period of 21 days and the number of parent  immobilised/dead organisms and/or possible abnormal behaviours both in control and in treatment group vessel and the total number of living offsrping produced per parent animal aliveat the end of the test have been observed. The presence of aborted eggs or dead offspring have been recorded. After a range finding test, a limit test has been performed to demonstrate that there is no statistically significant effect at the limit concentration tested.In the limit definitive test the organisms have been exposed to 10 mg/lof the test item for a total period of 21 days. At the beginning and at 3 times/week (at every test medium renewal) dissolved oxygen and pH have been measured in control and in the tested concentration. Also water temperature was recorded. A preliminary test to evaluate the stability of the test sample has been performed by TOC analyser. Then one time for week TOC has been measured in a pool derived from vessels in the tested concentration. 1 g/l of active matter is 3.920g/l of test item. Under the study conditions (OECD 211) the substance has a LOEC and NOEC > 10 mg/l after 21 days.

 

 

 

Description of key information

Under the study conditions (OECD 211) the substance has a LOEC and NOEC > 10 mg/l after 21 days.

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Dose descriptor:

NOEC

Effect concentration:

> 10 mg/L

Additional information

A study was conducted to determine the effect on reproduction of the substance to Daphnia magna according to OECD Guideline 211, in compliance with GLP. Daphnia magna were exposed to a series of concentrations of the test substance for 21 days to find the limit concentration. Total organic carbon (TOC) concentrations were measured with a TOC analyser and the test substance was considered stable over the duration of the test. Daphnie have been kept under observation for a period of 21 days and the number of parent  immobilised/dead organisms and/or possible abnormal behaviours both in control and in treatment group vessel and the total number of living offsrping produced per parent animal aliveat the end of the test have been observed. The presence of aborted eggs or dead offspring have been recorded. After a range finding test, a limit test has been performed to demonstrate that there is no statistically significant effect at the limit concentration tested.In the limit definitive test the organisms have been exposed to 10 mg/lof the test item for a total period of 21 days. At the beginnng and at 3 times/week (at every test medium renewal) dissolved oxygen and pH have been measured in control and in the tested concentration. Also water temperature was recorded. A preliminary test to evaluate the stability of the test sample has been performed by TOC analyser. Then one time for week TOC has been measured in a pool derived from vessels in the tested concentration. 1 g/l of active matter is 3.920g/l of test item. Under the study conditions (OECD 211) the substance has a LOEC and NOEC > 10 mg/l after 21 days.