Esterification products of salicylic acid and C13-(branched) alcohols

Short-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

Feb 2018 to April 2018

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2018

Report date:

2018

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Version / remarks:

13 April 2004

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

tridecyl 2-hydroxybenzoate

EC Number:

947-975-7

Molecular formula:

C20H32O3

IUPAC Name:

tridecyl 2-hydroxybenzoate

Specific details on test material used for the study:

Test Substance Name - Dermol TDSA
Purity - UVCB
Storage on Receipt - Room Temperature (15 – 30°C)
Disposal - None hazardous
Hazard - None hazardous

Test substance details were supplied by the Sponsor. A Certificate of Analysis was supplied for the compound;this is included in the study report.

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

- Concentrations:
Dermol TDSA:WAFs prepared at loading rates of 1.0, 3.2, 10, 32 and 100 mg/L.
Control Test medium without test item or other additives

- Sampling method:
Samples for possible analysis were taken from all test concentrations and the control according to the schedule below. In addition, the glass wool was kept for possible analysis. The method of analysis is described in the appended Analytical Report (Appendix 3).
Frequency at t=0 h and t=48 h
Volume 2.0 mL from the approximate centre of the test vessels
Storage Samples were stored in a freezer (≤-15°C) until analysis at the analytical laboratory of the Test Facility.
At the end of the exposure period, the replicates were pooled at each concentration before sampling.
Additionally, reserve samples of 2.0 mL were taken for possible analysis. If not used, these samples were stored in a freezer (≤-15°C) for a maximum of three months after delivery of the draft report, pending on the decision of the sponsor for additional analysis.

- Sample storage conditions before analysis:
Preparation of test solutions started with loading rates individually prepared at 1.0, 10 and 100 mg/L in the combined limit/range-finding test and 1.0, 3.2, 10, 32 and 100 mg/L in the final test. A two-day period of magnetic stirring was applied to ensure maximum dissolution of the test item in medium. The obtained mixtures were allowed to settle for an overnight period, where after the aqueous Water Accommodated Fractions (WAFs) were collected by means of siphoning through glass wool. The obtained test solutions were clear and colorless at the end of the preparation procedure and used as test concentrations. All test solutions used in the combined limit/range-finding test and from the limit concentration of the final test were checked on the Tyndall effect using a Laser pen. No undissolved particles were observed in any of the test solutions, indicating that only the dissolved part of the test item was present.
Any residual volumes were discarded.

Test solutions

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
- Eluate:
- Differential loading:
- Controls:
- Chemical name of vehicle (organic solvent, emulsifier or dispersant):
- Concentration of vehicle in test medium (stock solution and final test solution(s) or suspension(s) including control(s)):
- Evidence of undissolved material (e.g. precipitate, surface film, etc.):

- Test System
Species: Daphnia magna (Crustacea, Cladocera) (Straus, 1820), at least third generation, obtained by a cyclical parthenogenesis under specified breeding conditions.
Source: In-house laboratory culture with a known history.
Reason for selection: This system has been selected as an internationally accepted invertebrate species.
Validity of batch: Daphnids originated from a healthy stock, 2nd to 5th brood, showing no signs of stress such as mortality >20%, presence of males, ephippia or discoloured animals and there was no delay in the production of the first brood.
Characteristics: For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.

- Breeding
Start of each batch: With newborn daphnids, i.e. less than 3 days old, by placing about 250 of them into 5 litres of medium in an all-glass culture vessel.
Maximum age of the cultures: 4 weeks
Renewal of the cultures: After 7 days of cultivation half of the medium twice a week.
Temperature of medium: 18-22°C
Feeding: Daily, a suspension of fresh water algae.
Medium: M7, as prescribed by Dr. Elendt-Schneider (Elendt, B.-P., 1990: Selenium deficiency in Crustacea. An ultrastructural approach to antennal damage in Daphnia magna Straus. Protoplasma 154, 25-33).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Justification for species other than prescribed by test guideline: This system has been selected as an internationally accepted invertebrate species.
- Source: In-house laboratory culture with a known history.
- Age of parental stock (mean and range, SD): For the test selection of young daphnids with an age of < 24 hours, from parental daphnids of more than two weeks old.
- Feeding during test: Yes
- Food type: A suspension of fresh water algae.
- Frequency: Daily

Study design

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Hardness:

Not reported

Test temperature:

18-22°C

pH:

6-9

Dissolved oxygen:

>=3mg/L at the end of the test

Salinity:

Not applicable

Conductivity:

Not reported.

Nominal and measured concentrations:

Range-finding test: WAFs prepared at loading rates of 1.0, 3.2, 10, 32 and 100 mg/L.
Definitive test: 10, 32,100 mg/l loading rate

Details on test conditions:

TEST SYSTEM
Test duration 48 hours
Test type Static
Test vessels 50 mL, all-glass
Medium Adjusted ISO medium
Number of daphnids 20 per concentration
Loading 5 per vessel containing 50 mL of test solution
Light 16 hours photoperiod daily
Feeding No feeding
Aeration No aeration of the test solutions.
Introduction of daphnids Within 30 minutes after preparation of the test solutions.


VEHICLE CONTROL PERFORMED: yes

RANGE-FINDING STUDY
The project started with a combined limit/range-finding test. Twenty daphnids per concentration (four replicates, 5 daphnids per vessel) were exposed to a control and a WAF prepared at a loading rate of 100 mg/L in the limit test. Test procedure and conditions were similar to those applied in the final test with the following exceptions:
• Ten daphnids per concentration (in duplicate, 5 per vessel) were exposed to WAFs prepared at loading rates of 1.0 and 10 mg/L in the combined range-finding test.
• Dissolved oxygen concentrations and pH were only measured in the control and the highest test concentration.

Reference substance (positive control):

yes

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

0.055 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: exposure concentration

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EL50

Effect conc.:

56 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: loading rate

Results with reference substance (positive control):

The objective of the study was to evaluate potassium dichromate (K2Cr2O7) for its ability to generate acute toxic effects on the mobility of Daphnia magna during an exposure period of 48 hours and, if possible, to determine the EC50 at 24 and 48 hours of exposure (Charles River Den Bosch Study Number 20143130).
The study procedures described in this report were based on the OECD guideline No. 202: "Daphnia sp., Acute Immobilisation Test", Adopted April 13, 2004 and the ISO International Standard 6341.
Start of first exposure: 02 Jan 2018
Completion last exposure: 04 Jan 2018
The reference test was carried out to check the sensitivity of the test system as used by Charles River Den Bosch. Daphnia were exposed for a maximum of 48 hours to K2Cr2O7 concentrations of 0.10, 0.18, 0.32, 0.56, 1.0 and 1.8 mg/L and to a control. Twenty daphnids were exposed per concentration.
The reference item, potassium dichromate (K2Cr2O7, art. 1.04864, batch no. K44879664) was obtained from Merck, Darmstadt, Germany.
Acute immobilization of daphnia after 24 and 48 hours in the reference test with potassium dichromate:
Concentration
K2Cr2O7 Number
Exposed % immobile
24h 48h Expected response (%)
After 48 hours 1
(mg/L) Minimal Maximal
control 20 0 0 0 102
0.10 20 0 0 0 10
0.18 20 0 0 0 10
0.32 213 0 5 0 30
0.56 20 0 55 0 100
1.0 20 80 100 40 100
1.8 20 100 100 100 100
1 Based on historical data of the previous years (n>60).
2 A maximum response of 10% does not invalidate the results of the test.
3 Unintentionally, an extra daphnid was added to one of the test vessels.

In conclusion, the actual responses in this reference test with K2Cr2O7 are within the ranges of the expected responses at the different concentrations, i.e. the 48h-EC50 was between 0.3 and 1.0 mg/L. Hence, the sensitivity of this batch of D. magna was in agreement with the historical data collected at Charles River Den Bosch.
The 24h-EC50 was 0.87 mg/L with a 95% confidence interval between 0.80 and 0.95 mg/L.
The 48h-EC50 was 0.54 mg/L with a 95% confidence interval between 0.49 and 0.61 mg/L.
The raw data from this study are kept in the Charles River Den Bosch archives. The test described above was performed under GLP with a QA-check.

Any other information on results incl. tables

Range-finding test results: Time (h) Replicate Dermol TDSA; Loading rate (mg/L)     Control 1 10 100 0 A 5 5 5 5   B 5 5 5 5   C 5 5   D 5 5   Total introduced 20 10 10 20 24 A 0 0 0 0(2)   B 0 0 0 0   C 0 2(5)   D 0 0(1)   Total immobilised 0 0 0 2   Effect % 0 0 0 10 48 A 0 0 3 5   B 0 0 4 5   C 0 4   D 0 5   Total immobilised 0 0 7 19   Effect % 0 0 70 95

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

In conclusion, the 48h-EC50 for Daphnia magna exposed to Dermol TDSA was 0.055 mg/L based on average exposure concentrations (95% confidence interval between 0.037 and 0.084 mg/L).
The 48h-EL50 for Daphnia magna exposed to Dermol TDSA was 56 mg/L based on loading rates (95% confidence interval between 40 and 80 mg/L).

Executive summary:

The objective of the study was to evaluate Dermol TDSA for its ability to generate acute toxic effects on the mobility of Daphnia magnaduring an exposure period of 48 hours and, if possible, to determine the EC₅₀at 24 and 48 hours of exposure.

The study procedures described in this report were based on the OECD guideline No. 202, 2004.In addition, procedures were based on the test methods described in the OECD series on testing and assessment number 23, 2000.

The batch of Dermol TDSA tested was a colourless to pale yellow clear liquid UVCB which was not completely soluble in test medium at the loading rates initially prepared. Water Accommodated Fractions (WAFs) were individually prepared at loading rates ranging from 1.0 to 100 mg/L and used as test concentrations.

A final test was performed based on the results of a preceding combined limit/range-finding test. Twenty daphnids per group (5 per replicate, quadruplicate) were exposed to an untreated control and to WAFs prepared at loading rates of 1.0, 3.2, 10, 32 and 100 mg/L. The total exposure period was 48 hours and samples for analytical confirmation of exposure concentrations were taken at the start and at the end of the test.

No immobility or other effects were observed in the control and at the lowest test concentration throughout the test. At the end of the test, 15, 5, 25 and 75% immobility was observed at the WAFs prepared at loading rates of 3.2, 10, 32 and 100 mg/L, respectively.

Samples taken from all concentrations and the control were analysed. The measured concentrations at the start of the test were below the Limit of Quantification (LOQ). No detectable test item was measured in the samples taken from the WAFs prepared at loading rates of 1.0 and 3.2 mg/L, already at the start of the test. Initial test item concentrations of 0.096, 0.11 and 0.22 mg/L could be estimated for the WAFs prepared at loading rates of 10, 32 and 100 mg/L, respectively, by extrapolation of the calibration curve. The estimated concentrations decreased below the Limit of Detection (LOD) during the 48-hour exposure period, except for the WAF prepared at a loading rate of 100 mg/L which was estimated to be 0.090 mg/L at the end of the test. It should be noted that the water solubility of the test item was determined to be <78 ng/L (Test Facility Study No. 20136818). It can be stated that testing was ultimately performed at the maximum soluble concentration of test item in test medium with an average exposure concentration of 0.14 mg/L.  

The study met the acceptability criteria prescribed by the study plan and was considered valid.

In conclusion, the 48h-EC₅₀forDaphnia magnaexposed to Dermol TDSA was 0.055 mg/L based on average exposure concentrations (95% confidence interval between 0.037 and 0.084 mg/L). The 48h-EL₅₀forDaphnia magnaexposed to Dermol TDSA was 56 mg/L based on loading rates (95% confidence interval between 40 and 80 mg/L).