Reaction products of n-octanol and acrylic acid, first distillation pitch

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:

From January 5, 2012 to January 27, 2012

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study was performed according to the OECD 203 guideline and EU C.1 Method with GLP.

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

Qualifier:

according to guideline

Guideline:

ISO 7346-1 (Determination of the Acute Lethal Toxicity of Substances to a Freshwater Fish [Brachydanio rerio Hamilton-Buchanan (Teleostei, Cyprinidae)] - Part 1: Static Method)

Deviations:

no

Qualifier:

according to guideline

Guideline:

other: Guidance document on aquatic toxicity testing of difficult substances and mixtures, OECD series on testing and assessment number 23, December 14, 2000.

Deviations:

no

GLP compliance:

yes (incl. QA statement)

Analytical monitoring:

yes

Details on sampling:

- Concentrations: all concentrations.
- Sampling method: Samples for Total Organic Carbon (TOC) analysis were controlled at 0 h and 96 h, with a volume of 40 mL.
- Sample storage conditions before analysis: Samples were stored in a refrigerator until analysis.

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method:
The test substance was not completely soluble in test medium at the loading rates initially prepared.
Preparation of test solutions started with loading rates ranging from 1.0 to 100 mg/L applying an overnight period of magnetic stirring to reach maximum solubility of the test substance in the test medium. The resulting aqueous mixtures were left to stabilize for 2 to 2¼ hours were after the Water Accommodated Fractions (WAFs) were taken out by siphoning through glass wool. The lowest test solution for the range-finding test was prepared by 10-fold dilution of the WAF prepared at a loading rate of 1.0 mg/L. All test solutions were clear and colourless besides the WAF prepared at 100 mg/L in the final test which was slightly hazy.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp.
- Source: Zodiac, proefacc, "De Haar Vissen", Wageningen University and Research Centre, The Netherlands.
- Length at study initiation: Range-finding test: 2.9 ± 0.2 cm, Final test: 3.0 ± 0.3 cm.
- Weight at study initiation (mean and range, SD): Range-finding test: 0.61 ± 0.17 g, Final test: 0.76 ± 0.21 g.
- Feeding during test: No feeding from 24 hours prior to the test and during the total test period.

ACCLIMATION
- Acclimation period: At least 12 days after delivery.

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

96 h

Hardness:

180 mg CaCO3 per litre.

Test temperature:

21.5 - 22.2 ºC

pH:

7.2 - 7.9

Dissolved oxygen:

5.7 - 9.4 mg/L

Nominal and measured concentrations:

WAFs prepared at loading rates of 10, 18, 32, 56 and 100 mg/L.

Details on test conditions:

TEST SYSTEM
- Test vessel: 20 litres, all-glass, containing 15 litres of test solution.
- Aeration: yes. Aeration was introduced after 48 hours of exposure and was terminated the next day.
- No. of organisms per vessel: 7
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Intervals of water quality measurement:
Dissolved oxygen content, pH and temperature: Daily in all vessels with surviving fish, beginning at the start of the test (day 0).

OTHER TEST CONDITIONS
- Photoperiod: 16 hours photoperiod daily

EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
Mortality and other effects: At 2¼, 24, 48, 72 and 96 hours following the start of exposure. In addition, every afternoon from day 0 and every morning from day 1 to observe for any dead or severely distressed fish. Dead fish were removed when observed.

TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 10% of 1.0 mg/L, 1.0 mg/L, 10 mg/L and 100 mg/L
- Results used to determine the conditions for the definitive study: The expected LL50 was between concentrations obtained in WAFs prepared at loading rates of 10 and 100 mg/L.

Reference substance (positive control):

yes

Remarks:

Penthachlorophenol (PCP)

Duration:

96 h

Dose descriptor:

LL50

Effect conc.:

56 - 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

-Mortality:
One fish died after 48 hour of exposure in the WAF prepared at 56 mg/L. All fish were found dead in the highest WAF after 24 hours of exposure.
At the end of the test all test solutions were observed to be hazy and the haziness increased with the loading rate, indicating that the test substance was not completely dissolved.
-Clinical effects:
There were no clinical effects observed in the control and WAFs prepared at 10, 18 and 32 mg/L during the test period.

Results with reference substance (positive control):

- Results with reference substance valid? Yes, the 96h-LC50 was 0.25 mg/L based on nominal concentrations, with a 95% confidence interval between 0.20 and 0.41 mg/L.

Sublethal observations / clinical signs:

Table 1. Incidence of mortality and total mortality during the final test.

Test substance^1 Loading rate^2 (mg/L)	Initial number of fish	Cumulativemortality					Total Mortality (%)
		2¼h	24h	48h	72h	96h
control	7	0	0	0	0	0	0
10	7	0	0	0	0	0	0
18	7	0	0	0	0	0	0
32	7	0	0	0	0	0	0
56	7	0	0	1	1	1	14
100	7	0	7	7	7	7	100

¹ Reaction products of n-octanol and acrylic acid, first distillation pitch.

² WAF prepared at given loading rate.

Table 2. Clinical effects observed during the final test.

Test substance^1 Loading rate^2 (mg/L)	Time of recording (hours)	Specification of effects	Relative number
56	24	Hypoactive Discoloured and loss of equilibrium	6/7 1/7
	48	Hypoactive Discoloured and loss of equilibrium Hypoactive and discoloured	1/7 1/7 5/7
	72	Hypoactive Hypoactive and discoloured	2/6 4/6
	96	Hypoactive	6/6
100	2¼	Hypoactive	7/7

¹ Reaction products of n-octanol and acrylic acid, first distillation pitch.

² WAF prepared at given loading rate.

Table 3. Effect parameters.

Parameter	Test substance^1 Loading rate^2 (mg/L)
NOELR	32
24, 48, 72 and 96h-LL50	Between 56 and 100

¹ Reaction products of n-octanol and acrylic acid, first distillation pitch.

² WAF prepared at given loading rate.

Validity criteria fulfilled:

yes

Remarks:

(no mortalitiy in the control; test conditions maintained during the test; dissolved oxygen > 5 mg/L)

Conclusions:

The 96h-LL50 was between WAFs prepared at loading rates of 56 and 100 mg/L.

Executive summary:

The 96-Hour Acute Toxicity Study in Carp with the test substance was carried out according to OECD 203 Guideline and EU C.1 Method (GLP study).

 

As the test substance was not completely soluble in test medium at the loading rates initially prepared, the method Water Accommodated Fractions (WAFs) was used. The concentration of test substance in the aquatic samples was calculated using the TOC content of pure test substance.

 

Reaction products of n-octanol and acrylic acid, first distillation pitch induced no mortality or clinical effects in carp at a WAF prepared at a loading rate of 32 mg/L (NOELR). The 96h-LL50was between WAFs prepared at loading rates of 56 and 100 mg/L.

Description of key information

Key study: Experimental results: OECD 203 guideline and EU C.1 Method. GLP study. 
The 96h-LL50 was between WAFs prepared at loading rates of 56 and 100 mg/L.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

56 mg/L

Additional information

Key study: Experimental results: OECD 203 guideline and EU C.1 Method. GLP study.

The 96h-LL50 was between WAFs prepared at loading rates of 56 and 100 mg/L.