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

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:

2018-03-26 - 2018-03-30

Reliability:

1 (reliable without restriction)

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: Ra-He 2014-054
- Expiration date of the lot/batch: 2018/10

STABILITY AND STORAGE CONDITIONS OF TEST MATERIAL
- Storage condition of test material: Keep container tightly closed and dry; store in a cool place
- Stability under test conditions: Stable
- Solubility and stability of the test substance in the solvent/vehicle: 63.35g/L in water

Type of the test substance: No Unique and identified molecular structure substance, 100% UVCB
Chemical name : Fatty acids, C12-14, reaction product with sulfur trioxide, sodium salt
CAS: 2215087-54-8

Analytical monitoring:

yes

Vehicle:

no

Test organisms (species):

other: Gobiocypris rarus

Details on test organisms:

The test was performed with Gobiocypris rarus which were bred in Bioassay and Safety Assessment Lab, Shanghai Academy of Public Measurement (ID: GR-SAPM-170110). This freshwater fish species is suitable for fish toxicity testing because of their ease of culturing and handling, sensitivity to a variety of chemical substances, and the extensive data base for this species. This species complies with “Ministry of environmental protection of the people's Republic of China, The Guidelines for the Testing of Chemicals–Effects on Biotic System, 2013, 203 Fish, Acute Toxicity Test”, “OECD, Guidelines for the Testing of Chemicals. 203 Fish, Acute Toxicity Test, 1992” and “GB/T 29763-2013 Chemicals-Rare Minnow (Gobiocypris rarus) acute toxicity test”.

Before being used in the test, all fish were acclimatized in the following test conditions for more than 14 days:
Water: the test water used was tap water dechlorinated by activated carbon and sterilized by UV lamp; Temperature: 21-25℃ and constant within a range of 2°C, pH: 6.0-8.5;
Dissolved oxygen concentration: > 80% of air saturation value (ASV);
Photoperiod: Light/Dark = 12 hours/12 hours;
Feeding: fish were fed with brine shrimp flake (O.S.I®, USA) daily until 24 hours before the test was started.
Length was 2.5±0.1cm (Mean ± SD) and weight was 0.26±0.02g (Mean ± SD) at the end of the test. The age of the test fish was approximately 13 months old, before the test they were not medically treated.
No diseases were observed during the acclimation period;

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Details on test conditions:

Duration: 96h;
Test method: static for range-finding test, and semi-static for definitive test and its renewal frequency was 48h;
Loading: about 0.5g fish/litre;
Temperature: 21.5-22.6°C;
Photoperiod: Light/Dark=12 hours/12 hours;
Dissolved oxygen concentration: 80.4%-95.3%ASV;
Feeding: none.

Three loading rates of 1.2, 12 and 120mg/L were used in the range-finding test. In this test, a blank control of test water was used. Five fish were introduced into each vessel and the test duration was 96 hours. A static method was used in this test.
Observation and measurements
Fish were observed and dead fish were recorded at 0, 24, 48, 72 and 96 hours after test initiation. For the analysis of the actual test substance concentration, two 2mL of test solution was taken from each test vessel at the beginning, 24h, 48h and at the end of the test.

Test solutions preparation - Definitive test
The test solutions were prepared by adding the required amout of the test substance into 6L test water and stirring for about 30min. The vortex depth was 1/10 of the mixture column. After the stirring period, the test substance was completely dissovled by visual inspection. Then the mixtures were settled for about one hour and the middle of the mixtures (thefirst 100mL were discarded) were used as the test solutions. The WAFs at each loading were prepared individually.

Procedure
Based on the range-finding test, five treatment groups with loading rates of 1.2mg/L, 2.2mg/L, 4mg/L, 7mg/L and 12mg/L were used in the definitive test. In this test, a blank control of test water was used. Seven fish were introduced into each vessel and the test duration was 96 hours. A 48h semi-static method was used in this test.

Observation and measurements
The fish were inspected at 0, 3, 24, 48, 72 and 96 hours after test initiation and dead fish were removed when observed and mortalities and any abnormalities observed were recorded. Measurements of the pH, dissolved oxygen concentration and temperature in the test solutions were made at the beginning, 24h, before and after 48h renewal and at the end of the test . For the analysis of the actual test substance concentration, 2mL of test solution was taken from each test vessel at the beginning, before and after 48h renewal, and at the end of the test. At the end of the test, the body length and body weight of the test fish of the control group were measured. The loading rate was also calculated with the body weight at the same time.

Statistical analysis
The mortalities were plotted against each test loading and control for each exposure period (24h, 48h, 72h and 96h). The 24 hour LL50 with 95% confidence limits were calculated using the method of Maximum Likelihood-Probit. The 48, 72 and 96 hours LL50 with 95% confidence limits were calculated using the method of Spearman-Karber. The data analysis was carried out with software TOXCALC-Toxicity Data Analysis Software (v5.0.32).

Method validation
No mortality was observed in the control during the test;
Acceptable exposure conditions were maintained throughout the test. The dissolved oxygen concentrations in test solutions were between 80.4%- 95.3%ASV, which was greater than 60.0% of the air saturation value, throughout the test. During the test, the measured concentrations of peak 1 and peak 2 in each group varied less than 20% during each renewal period (-13.8% to 4.35%). The test fulfilled the above requirements and was therefore considered to be valid.

Conclusions
The 96h LL50 value for Texapon SFA to Gobiocypris rarus was 5.72mg/L and 95 percent confidence limits ranged from 4.95mg/L to 6.62mg/L.
The maximum loading rate causing no mortality and the minimum loading rate causing 100 percent mortality within the period of the test was 4mg/L and 12mg/L, respectively. The results above were expressed relative to the loading rate of the test substance. The analysis of the two main components confirmed that the concentrations were kept stable during the test.

Key result

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

5.72 mg/L

Nominal / measured:

nominal

Conc. based on:

act. ingr.

Basis for effect:

mortality (fish)

Reported statistics and error estimates:

The mortalities were plotted against each test loading and control for each exposure period (24h, 48h, 72h and 96h). The 24 hour LL50 with 95% confidence limits were calculated using the method of Maximum Likelihood-Probit. The 48, 72 and 96 hours LL50 with 95% confidence limits were calculated using the method of Spearman-Karber. The data analysis was carried out with software TOXCALC-Toxicity Data Analysis Software (v5.0.32).

Sublethal observations / clinical signs:

The measured concentrations of peak 1 and peak 2 in the test substance in definitive test

Group			Blank Control		Treatment group (Loading rate, mg/L)
					1.2		2.2		4		7		12
			peak1	Peak2	peak1	Peak2	peak1	Peak2	peak1	Peak2	peak1	Peak2	peak1	Peak2
Measured concentration (mg/L)	0		ND3	ND4	0.754	0.250	1.43	0.622	2.80	1.10	4.93	2.00	7.75	3.34
	48	O	ND3	ND4	0.784	0.251	1.49	0.622	2.82	1.15	5.08	2.09	8.00	3.25
		N	ND3	ND4	0.858	0.372	1.52	0.664	2.81	1.16	4.96	2.20	/	/
	96		ND3	ND4	0.836	0.327	1.54	0.628	2.90	1.15	5.17	2.15	/	/
Geometric mean (mg/L)			-	-	0.807	0.296	1.49	0.634	2.83	1.14	5.03	2.11	7.87	3.29
48h-O vs. 0h(%)			-	-	3.83	0.398	4.03	0.00	0.709	4.35	2.95	4.31	3.13	-2.77
96h vs.48h-N(%)			-	-	-2.63	-13.8	1.30	-5.73	3.10	-0.870	4.06	-2.33	-	-

ND3/ND4: not detected, LOD3/LOD4=127ng/mL /106ng/mL . “-”, not available. “O”, before the renewal. “N”, after the renewal.“/”, not measured.

Description of key information

The 96-h LC50 value of the key study is 5.72 mg/L in freshwater fish in chinese rare minnow (Gobiocypris rarus)

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Dose descriptor:

LC50

Effect concentration:

5.72 mg/L

Additional information

 

Two short term toxicity tests on fish are available:

 

The short-term toxicity to freshwater fish was determined in a study according to OECD TG 203 and in compliance with GLP criteria. In this study, groups of 7 zebra fish (D. rerio) the 96-h LL50 was calculated as geometric mean of the LL0 and LL100 to be 17.7 mg/L. In this study all OECD validity criteria were met.

In a second study which is used for PNEC derivation, Chinses rare minnow (Gobiocypris rarus) were exposed for 96 h towards the registered test substance according to OECD 203 (GLP study). The LC50 was determined to be 5.73 mg/L. In this study all OECD validity criteria were met.