Fatty acids, C12-14, .alpha.-sulfo, disodium salts

Long-term toxicity to fish

Link to relevant study record(s)

Reference

Endpoint:

fish early-life stage toxicity

Type of information:

experimental study

Adequacy of study:

key study

Study period:

2021-10-26 - 2021-12-30

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test)

GLP compliance:

yes

Specific details on test material used for the study:

Name Fatty acids, C12-14, reaction product with sulfur trioxide, sodium salt
Type of the test substance No unique and identified molecular structure substance, UVCB

CAS No 2215087-54-8
Batch No 0021197214
Appearance White solid
Odour Characteristic
Density 1.3074 (20 °C)
Solubility in water 63.35 g/L

Analytical monitoring:

yes

Remarks:

UPLC-MS analysis method

Vehicle:

no

Details on test solutions:

13.2.1 Test solutions preparation
Based on the information provided by the sponsor, the test substance is identified as an UVCB and the solubility in water is 63.35 g/L. Thus, the test solutions were prepared as water accommodated fraction (WAFs). WAFs were prepared by adding the required amount of the test substance into a certain amount of water and stirring until dissolved. The vortex depth was 1/10 of the test solution column. Each test solution used in the test was prepared as loading with a specified concentration of the test substance in the test water during the stirring period. At the end of the stirring period, the mixtures were settled for about one hour and then the middle of the mixtures (the first 100 mL were discarded) was used as the test solutions. The test solutions were visually checked for undissolved test substance before use.
13.2.2 Procedure
Based on the result of an acute fish toxicity test, five loading rates of 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L were used in the Early_life Stage Toxicity test (arranged in a geometric series with a separation factor of 2.65). In this test, a blank control of test water was used. The treatment and control groups consisted of four replicates with 80 embryos. The test duration was 35 days and the frequency of renewal was every 48h.

Four major components peaks were detected by UPLC-MS in the treatment groups.
Only for the treatment group with a loading rate of 5.7 mg/L , peak 2 and peak 4 could be detected. For all other treatment groups the peaks 2 and 4 were below the detection limit (LOD). The LOD for peak 2 is 0.127 mg/L and for peak 4 the LOD is 0.0119 mg/L.
Peak 1 and peak 3 could be detected for all treatment groups and are expressed as time weighted arithmetic means of:

loading rate mean measured concentration
0.115 mg/L 0.091 mg/L with peak 1: 0.0607mg/L peak 3: 0.0303 mg/L
0.306 mg/L 0.1522 mg/L with peak 1: 0.09 mg/L peak 3: 0.0622 mg/L
0.811 mg/L 0.359 mg/L with peak1: 0.211 mg/L peak 3: 0.148 mg/L
2.15 mg/L 1.006 mg/L with peak 1: 0.6 mg/L peak3: 0.406 mg/L
5.7 mg/L 3.6 mg/L with peak 1: 2.1 mg/L, peak 2: 0.179 mg/L,
peak 3: 1.14 mg/L, peak 4: 0.180 mg/L

Test organisms (species):

Danio rerio (previous name: Brachydanio rerio)

Details on test organisms:

The test was performed with Danio rerio embryos (ID: DR-SAPM-20211125), while their parents were bred in Bioassay and Safety Assessment Lab, Shanghai Academy of Public Measurement.
Before egg production for this test, the parent fish used in the test were acclimated.

Food: fish were fed with brine shrip flake and live food of brine shrimp, O.S.I®.
Feeding: fish were fed twice a day.
Length of the fish hatching in the control group for each replicate were 1.1cm±0.1cm; 1.2cm±0.2cm; 1.1cm±0.1cm and 1.2cm±0.2cm（Mean±SD）and fresh weight were 0.0243g±0.0042g; 0.0246g±0.0045g; 0.0234g±0.0027g and 0.0229g±0.0039g (Mean ± SD) at the end of the test.

Embryos collected for use in the test were from forty individual (i.e. twenty females and twenty males) spawns and the test was started within 2 hours after the eggs were fertilised. The embryos were removed from the spawning substrates and examined under a dissecting microscope to select healthy, viable specimens at a 8-cell stage About 100 intact fertilized egges were chosen and randomly distributed into each container (with the volume of 1000mL), at least six containers will be used in this test (at least five for treatment group with different test solutions, and one for blank control). The embryos were immersed in the test solutions before cleavage of the blastodisc commences. Then, about 80 of the 100 eggs were selected randomly for each test group, 20 fertilized eggs were be distributed randomly to each replicate (the order of distribution followed a the random number generated by Excel). The embryos used in the test were distributed at simple random and unproperly treated embryos were discarded.The test duration was 35 days, 30 days after all viable embryos in blank control group had hatched (day 5).

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

35 d

Test temperature:

24.7-25.9°C

pH:

6.0 - 8.5

Nominal and measured concentrations:

loading rates: 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L corresponding to mean measured concentration of 0.091 mg/L, 0.1522 mg/L, 0.0359 mg/L, 1.006 mg/L and 3.6 mg/L

Details on test conditions:

13.1 Test conditions
13.1.1 Water
Standard dilution water was used. The water was aerated continuously over 24h.
The water was aerated continuously over 24h. Water quality parameters were as follows:
Total hardness: 150mg CaCO3/L;
Oxygen concentration: >90%ASV;
Full details of water quality parameters are given in Appendix 4.
13.1.2 Test vessels
250mL self-made glass test chambers (9 cm in diameter) containing 200mL test solution at 0-15d; 2000mL self-made glass test chambers (15 cm in diameter) containing 1500mL test solution at 16-35d .
13.1.3 Conditions of exposure
Duration: Within 2 hours post-fertilization (i.e. 8-cell stage) at test start and about 30-Day Post-Hatch of the control group (35d in total);
Test method: semi-static, the frequency of renewal was every 48h;
Loading: about 0.317g fish/L pre 24h (based on the average of the fresh weight in control at the end of the test);
Temperature: 24.7-25.9°C;
Photoperiod: Light/Dark=12 hours/12 hours;
Oxygen concentration: 72.5%-98.8%ASV;
Feeding: newly bred fishes were fed with live food of brine shrimp (Ocean Star International(O.S.I), Inc,USA), and the frequency was twice a day and two feeds were separated by 6 hours. The quantity of food given to fish was constant in each test group.
Cleaning: the test vessel was replaced by a clean one each time the test solution was renewed.

13.2.3 Observation and measurements
The fish were inspected daily and for hatching, mortality and other observed effects and the findings were recorded. Dead embryos and fish were removed when observed.
All survivors were treated by anaesthesia with 80mg/L MS-222 at the end of the test. The wet weights (blotted dry) and lengths were measured individually. Fish were not be weighed during the test.
Total hardness was measured in the control and the highest concentration at he begining and the end of the test.
The pH, dissolved oxygen (DO) concentration and temperature were measured in the test solutions of all replicates at the beginning, before and after each renewal and at the end of the test. Additionally, the temperature was monitored continuously in one test vessel of the control group).
For analysis of the concentration, 20mL of test solution was taken from the test vessel which was selected randomly from each group at the beginning, before and after each renewal and at the end of the test. Alternating replicates were used for weekly samplings.

Key result

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

1.006 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

length

Key result

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

2.15 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

length

Key result

Duration:

35 d

Dose descriptor:

NOEC

Effect conc.:

1.006 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

weight

Key result

Duration:

35 d

Dose descriptor:

NOELR

Effect conc.:

2.15 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

weight

Key result

Duration:

35 d

Dose descriptor:

NOELR

Effect conc.:

1.006 mg/L

Nominal / measured:

meas. (arithm. mean)

Conc. based on:

act. ingr.

Basis for effect:

mortality

Key result

Duration:

35 d

Dose descriptor:

NOELR

Effect conc.:

2.15 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality

Details on results:

Under the test conditions, all the embryos in the treatment and control groups were hatched within 6 days. The difference of hatching rates of fertilized eggs between the treatment groups and the blank control was not statistically significant (p=0.449，p＞0.05).
All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L.
No statistical significance up to the concentration of 2.15 mg/L could be detected for mortality at the end of exposure (day 35). The NOELR is determined to be 2.15 mg/L.corresponding to 1.006 mg/L (mean meassured) and representing the highest concentration with < 5% post- hatch mortality. (Since the study report evaluates mortality by the indication to be "< 5%", the NOELR for mortality was re-evaluated in a separate non-GLP statistical evaluation.)
For fresh weight and length at the end of the test the no observed effect loading rate (NOELR) was 2.15 mg/L corresponding to 1.006 mg/L (mean measured). The result above were expressed relative to the loading rates of the test solutions.

Validity criteria fulfilled:

yes

Conclusions:

In a new guideline study according to OECD 210 the aquatic toxicity of the registered substance (UVCB) was tested in zebra fish (Danio rerio). The study was performed in a semi static approach with a renewal of the test concentration every 48 h over a test duration of 35 days.

Five concentrations were selected with loading rates of 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L corresponding to mean measured concentration of 0.091 mg/L, 0.1522 mg/L, 0.0359 mg/L, 1.006 mg/L and 3.6 mg/L.

All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L.
No statistical significance up to the concentration of 2.15 mg/L could be detected for mortality at the end of exposure (day 35). The NOELR is determined to be 2.15 mg/L.corresponding to 1.006 mg/L (mean meassured) and representing the highest concentration with < 5% post- hatch mortality.
For fresh weight and length at the end of the test the no observed effect loading rate (NOELR) was 2.15 mg/L corresponding to 1.006 mg/L (mean measured). The result above were expressed relative to the loading rates of the test solutions.

Executive summary:

In a new guideline study according to OECD 210 the aquatic toxicity of the registered substance (UVCB) was tested in zebra fish (Danio rerio). The study was performed in a semi static approach with a renewal of the test concentration every 48 h over a test duration of 35 days.

Five concentrations were selected with loading rates of 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L corresponding to mean measured concentration of 0.091 mg/L, 0.1522 mg/L, 0.0359 mg/L, 1.006 mg/L and 3.6 mg/L.

All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L.

In a new guideline study according to OECD 210 the aquatic toxicity of the registered substance (UVCB) was tested in zebra fish (Danio rerio). The study was performed in a semi static approach with a renewal of the test concentration every 48 h over a test duration of 35 days.

Five concentrations were selected with loading rates of 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L corresponding to mean measured concentration of 0.091 mg/L, 0.1522 mg/L, 0.0359 mg/L, 1.006 mg/L and 3.6 mg/L.

All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L.
The NOELR for mortality (post hatch), weight and length is determined to be 2.15 mg/L.corresponding to 1.006 mg/L (mean meassured) . The result above were expressed relative to the loading rates of the test solutions.

The result above were expressed relative to the loading rates of the test solutions.

 

 

Description of key information

In a new guideline study according to OECD 210 the aquatic toxicity of the registered substance (UVCB) was tested in zebra fish (Danio rerio). All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L. The NOELR  for mortality (post hatch), weight and length is 2.15 mg/Lcorresponding to 1.006 mg/L (mean meassured).The result above were expressed relative to the loading rates of the test solutions.

 

 

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

NOEC

Effect concentration:

1.006 mg/L

Additional information

In a new guideline study according to OECD 210 the aquatic toxicity of the registered substance (UVCB) was tested in zebra fish (Danio rerio). The study was performed in a semi static approach with a renewal of the test concentration every 48 h over a test duration of 35 days.

Five concentrations were selected with loading rates of 0.115 mg/L, 0.306 mg/L, 0.811 mg/L, 2.15 mg/L and 5.70 mg/L corresponding to mean measured concentration of 0.091 mg/L, 0.1522 mg/L, 0.0359 mg/L, 1.006 mg/L and 3.6 mg/L.

All test organisms of the treatment group of the highest loading rate died within 3 days after hatch. The 35d median lethal loading rate (35d-LL50) was 3.03 mg/L.
No statistical significance up to the concentration of 2.15 mg/L could be detected for mortality at the end of exposure (day 35). The NOELR for mortality  (post hatch) weight and length is determined to be 2.15 mg/L.corresponding to 1.006 mg/L (mean meassured) . The result above were expressed relative to the loading rates of the test solutions.