Soybean oil, maleated, ester with triethanolamine

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:

Experimental start date: 12 June 2017 Experimental completion date: 30 June 2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

Deviations:

no

Qualifier:

according to guideline

Guideline:

EU Method C.1 (Acute Toxicity for Fish)

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

Physical state/Appearance: Clear amber viscous liquid
Purity: 100% product
Expiry Date: 24 June 2018
Storage Conditions: Room temperature, in the dark

Analytical monitoring:

yes

Details on sampling:

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 Water Accommodated Fraction (WAF) of the test item.
A nominal amount of test item (2200 mg) was added to the surface of 22 liters of test water to give the 100 mg/L loading rate.

Vehicle:

no

Remarks:

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 Water Accommodated Fraction (WAF) of the test item.

Details on test solutions:

A nominal amount of test item (2200 mg) was added to the surface of 22 liters of test water to give the 100 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. No particles or micro-dispersions of test item were observed by microscopic observations, however, the water column of the 100 mg/L loading rate was a cloudy white emulsion and hence it was considered justifiable to remove the WAF by filtering through a glass wool plug (2-4 cm in length) and filter paper. 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 Tygon tubing was inserted into the glass tube and pushed through the Nescofilm seal. A glass wool plug was inserted into the opposite end of the tubing and the WAF removed by mid-depth siphoning (the first 75-100 mL discarded) before passing through a filter paper to give the 100 mg/L loading rate WAF. Microscopic observations were performed on the WAFs after filtering and showed no particles or micro- dispersions of test item to be present, however, the preparations were observed to remain cloudy.

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

The test was carried out using juvenile rainbow trout (Oncorhynchus mykiss). Fish were obtained from Brow Well Fisheries Limited, Hebden, near Skipton, Yorkshire, UK and maintained in-house since 31 May 2017. Fish were maintained in a glass fiber tank with a "single pass" water renewal system. Fish were acclimatized to test conditions from 19 June 2017 to 26 June 2017. The lighting cycle was controlled to give a 16 hours light and 8 hours darkness cycle with 20 minute dawn and dusk transition periods.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

14 to 15 ºC

pH:

Between 7.8 @ 8.2

Dissolved oxygen:

dissolved oxygen content of greater than or equal to 9.3 mg O2/L

Salinity:

N/A

Nominal and measured concentrations:

A "Limit test" was conducted at a single loading rate of 100 mg/L

Reference substance (positive control):

no

Key result

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

> 100 mg/L

Nominal / measured:

not specified

Conc. based on:

other: loading rate WAF

Basis for effect:

mortality (fish)

Remarks on result:

other: 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Key result

Duration:

96 h

Dose descriptor:

other: The No Observed Effect Loading rate

Effect conc.:

100 mg/L

Nominal / measured:

not specified

Conc. based on:

other: loading rate WAF

Basis for effect:

mortality (fish)

Remarks on result:

other: The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Details on results:

The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Sublethal observations / clinical signs:

Cumulative mortality data from the exposure of rainbow trout to the test item during the range-finding test are given in Table 1.

The results showed no mortalities at 1.0, 10 and 100 mg/L loading rate WAF, additionally, there were no sub-lethal effects of exposure observed during the range-finding test.

Based on this information, a single loading rate of 100 mg/L was selected for the definitive test. This experimental design conforms to a "Limit test" to confirm that no mortalities or sub-lethal effects of exposure were observed.

 

Definitive Test

 

Total Organic Carbon (TOC) analysis of the fresh test preparations at 0 and 72 hours (see Annex 3) showed measured concentrations of 66 and 102 mg Carbon/L. TOC analysis of the old test solutions at 24 and 96 hours showed measured concentrations of 56 and 19 mg Carbon/L.

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

Cumulative mortality data from the exposure of rainbow trout to the test item during the definitive test are given in Table 2.

There were no mortalities in 7 fish exposed to a 100 mg/L loading rate WAF for a period of 96 hours. Inspection of the mortality data gave the following results:

 

 

Time (h)                             LL50 *

(mg/L Loading Rate WAF)

1                                            >100

3                                            >100

6                                            >100

24                                         >100

48                                         >100

72                                         >100

96                                        >100

* 95% confidence limits could not be calculated

 It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L.

 

There were no sub-lethal effects of exposure observed in 7 fish exposed to a 100 mg/L loading rate WAF for a period of 96 hours.

 

The test was considered to be valid given that none of the control fish died or showed signs of stress during the test and that the oxygen concentration at the end of the test was greater than or equal to 60% of ASV (6.1 mg O2/L) in the control and test vessels.

During the range-finding test, after 48 hours of exposure, the dissolved oxygen concentration in the 100 mg/L test media was measured at 4.4 mg O2/L and therefore below the 60% ASV (6.1 mg O2/L) validity criteria, as a result the aeration rate was increased. Given that no mortalities or sub-lethal effects were observed the reduced dissolved oxygen concentration was considered not to have adversely affected the results of the test.

The results of the water quality measurements are given in Table 3. Temperature was maintained at 14 Deg C to 15 Deg C throughout the test, while there were no treatment related differences for oxygen concentration or pH.

The vortex depth was recorded at the start and end of each mixing period and was observed to be a dimple at the water surface on each occasion.

Observations on the test media were carried out during the mixing and testing of the WAF.

At the start of each mixing period the 100 mg/L loading rate was observed to be clear colorless water column with test item floating at the water surface. After 23 hours stirring and a 1-Hour standing period the 100 mg/L loading rate was observed as a cloudy white water column. Microscopic inspection of the WAF showed no micro-dispersions or undissolved test item to be present, however, given that the test item had formed an emulsion the WAF was filtered through a 2-4 cm glass wool plug and filter paper. Microscopic examination after filtering showed no particles or micro-dispersions of the test item to be present, however, the test solution was observed to remain cloudy in appearance. During the test the 100 mg/L loading rate was observed to be a cloudy emulsion.

Table 1            Cumulative Mortality Data in the Range-finding Test

 

Nominal Loading Rate (mg/L)	Cumulative Mortality (Initial Population = 3)
	1 Hour	3 Hours	6 Hours	24 Hours	48 Hours	72 Hours	91 Hours
Control	0	0	0	0	0	0	0
1.0	0	0	0	0	0	0	0
10	0	0	0	0	0	0	0
100	0	0	0	0	0	0	0

Table 2            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
100	0	0	0	0	0	0	0	0

Table 3            Water Quality Measurements

 

Nominal Loading Rate (mg/L)	Time (Hours)
	0 Hours (Fresh Media)			24 Hours (Old Media)
	pH	mg O2/L	T ºC	pH	mg O2/L	T°C
Control	7.8	9.8	15	7.9	9.2	15
100	7.8	9.9	15	8.0	9.3	15

 

Nominal Loading Rate (mg/L)	Time (Hours)
	24 Hours (Fresh Media)			48 Hours (Old Media)
	pH	mg O2/L	T ºC	pH	mg O2/L	T°C
Control	7.9	9.8	15	8.3	9.5	15
100	7.9	9.9	15	8.2	9.1	15

 

Nominal Loading Rate (mg/L)	Time (Hours)
	48 Hours (Fresh Media)			72 Hours (Old Media)
	pH	mg O2/L	T ºC	pH	mg O2/L	T°C
Control	7.7	9.9	15	7.9	9.2	15
100	7.8	9.8	14	7.9	7.6	15

 

Nominal LoadingRate (mg/L)	Time (Hours)
	72 Hours (Fresh Media)			96 Hours (Old Media)
	pH	mg O2/L	T ºC	pH	mg O2/L	T°C
Control	8.0	9.7	15	8.1	9.7	15
100	7.9	9.6	15	8.0	8.9	15

Annex 2           Typical Water Quality Characteristics

 

Typical Water Characteristics for Supply Zone ZDB13

 

Analysis	Typical Value	Units
Hardness Level	Slightly Hard	N/A
Hardness clark	8.76	Degrees clark
Hardness french	12.52	French degrees
Hardness german	7.01	German degrees
Aluminium	7.46	mg Al/L
Chlorine	0.32	mg/L
Coliformbacteria	0	no./100mL
Colour	0.6	mg/l Pt/Co
Conductivity	345	μS/cm at 20°C
E.Colibacteria	0	no./100mL
Fluoride	0.24	mg F/L
Iron	19.9	μg Fe/L
Manganese	2.9	μg Mn/L
Nitrate	7.61	mg NO3/L
Odour	0	Dilution number
Pesticides	0	μg/L
pH	7.66	pH Value
Sodium	22.35	mg Na/L
Taste	0	Dilution number
Plumbing Metals
Copper	0.02	mg Cu/L
Lead	0.51	μg Pb/L
Nickel	3.1	μg Ni/L

 

Source:https://www.stwater.co.uk/my-supply/water-quality/check-my-water-quality/

Annex 3               Total Organic Carbon Analysis

 

METHOD OF ANALYSIS

Summary

The concentration of Total Organic Carbon (TOC) in the WAF was determined by TOC analysis.

Samples

In the range-finding test, samples of the control and each loading rate were taken for analysis at 0 hours (fresh media) and 24 hours (old media).

 

In the definitive test, samples of the control and 100 mg/L loading rate WAF were taken for analysis at 0 and 72 hours (fresh media), 24 and 96 hours (old media).

Standards

Standard solutions of potassium hydrogen phthalate (C8H5KO4) were prepared in reverse osmosis water at concentrations of 10, 20, 50 and 100 mg Total Carbon/L. Standard solutions of sodium carbonate (Na2CO3) were prepared in reverse osmosis water at concentrations of 5, 10, 20 and 33.33 mg Inorganic Carbon/L. These solutions were used for calibration of the carbon analyzer.

Procedure

The standard and sample solutions were analyzed for TOC using a Shimadzu TOC-VCPH high temperature Total Organic Carbon Analyzer using the following conditions:

 

Total Carbon Channel

Temperature                   :              680 ºC

Carrier gas                         :              zero grade air

Carrier gas flow rate      :              150 cc/min regulated at 50 psi

Catalyst                               :              platinum based

Injection volume             :              50 µL

 

Inorganic Carbon Channel

Temperature                   :              ambient

Carrier gas                         :              zero grade air

Carrier gas flow rate      :              150 cc/min regulated at 50 psi Catalyst :              25% orthophosphoric acid

Injection volume             :              50 µL

RESULTS

 

Range-finding Test

Samples	Nominal Loading Rate (mg/L)	Concentration of TOC (mg C/L)	Concentration of TOC Corrected for Control (mg C/L)
0 Hours (Fresh Media)	Control	1.34	-
	1.0	2.03	0.69
	10	3.13	1.8
	100	22.40	21
24 Hours (Old Media)	Control	<LOQ	-
	1.0	<LOQ	<LOQ
	10	3.04	3.0
	100	21.65	22

 

DefinitiveTest

Samples	Nominal Loading Rate (mg/L)	Concentration of TOC (mg C/L)	Concentration of TOC Corrected for Control (mg C/L)
0 Hours (Fresh Media)	Control	<LOQ	-
	100	66.00	66
24 Hours (Old Media)	Control	<LOQ	-
	100	56.38	56
72 Hours (Fresh Media)	Control	<LOQ	-
	100	102.3	102
96 Hours (Old Media)	Control	<LOQ	-
	100	19.26	19

 

LOQ = Limit of quantification which was considered to be 1.0 mg C/L

 

 

 

 

 

 

Validity criteria fulfilled:

yes

Conclusions:

The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Executive summary:

A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

 

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).

 

Following a preliminary range-finding test, seven fish were exposed to a Water Accommodated Fraction (WAF) of the test item, at a single nominal loading rate of 100 mg/L for a period of 96 hours at a temperature of 14 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.

 

Total Organic Carbon (TOC) analysis of the fresh test media at 0 and 72 hours showed measured concentrations of 66 and 102 mg/L respectively. TOC analysis of the old test media at 24 and 96 hours showed measured concentrations of 56 and 19 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 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

 

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L loading rate WAF.

Description of key information

The acute toxicity of the test item to the freshwater fish rainbow trout (Oncorhynchus mykiss) has been investigated and gave a 96-Hour LL50 value of greater than 100 mg/L loading rate WAF. The No Observed Effect Loading rate was 100 mg/L loading rate WAF.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

100 mg/L

Additional information

A study was performed to assess the acute toxicity of the test item to rainbow trout (Oncorhynchus mykiss). The method followed was designed to be compatible with the OECD Guidelines for Testing of Chemicals (1992) No 203, "Fish, Acute Toxicity Test" referenced as Method C.1 of Commission Regulation (EC) No. 440/2008.

 

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).

 

Following a preliminary range-finding test, seven fish were exposed to a Water Accommodated Fraction (WAF) of the test item, at a single nominal loading rate of 100 mg/L for a period of 96 hours at a temperature of 14 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.

 

Total Organic Carbon (TOC) analysis of the fresh test media at 0 and 72 hours showed measured concentrations of 66 and 102 mg/L respectively. TOC analysis of the old test media at 24 and 96 hours showed measured concentrations of 56 and 19 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 100 mg/L loading rate WAF. The No Observed Effect Loading Rate was 100 mg/L loading rate WAF.

 

It was considered unnecessary and unrealistic to test at loading rates in excess of 100 mg/L loading rate WAF.