3-hydroxypropiononitrile

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:

2008-09-23 to 2008-09-25

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: Guideline study OECD 202, Limit-test, GLP.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

2008

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

OECD Guideline 202 (Daphnia sp. Acute Immobilisation Test)

Deviations:

no

Principles of method if other than guideline:

A preliminary range finding test was conducted with three test item concentrations in the range of 10 to 1000 with a dilution factor of 10 under static conditions (NON-GLP-state).

Immobilization Rates in the Preliminary Test in [%] (n = 20)
Nominal
Test Item Concentration [mg/L] IMMOBILIZATION [%]
------------------------------------------------------------------------------------
24 h 48 h
Replicates Replicates
1 2 MV 1 2 MV
----------------------------------------------------------------------------------------
1000 0 0 0 0 0 0
100 0 0 0 0 0 0
10 0 0 0 0 0 0
Control 0 0 0 0 0 0
-----------------------------------------------------------------------------------------
MV = Mean value

Based on the results of the preliminary test the definitive test will be performed as a limit test with the nominal test item concentration 100 mg/L
under static conditions to enable the determination of immobilization after 24 and 48 hours. 2 to 24 h old daphnids will be used for the definitive
study. They will be obtained by removing the mother animals twice within 22 h. The daphnids will be acclimated to the dilution water at least 2 hours
before test start. The daphnids will not be fed during the study. Control replicates with dilution water (test medium without test item) will be tested
under the same conditions as the test groups. 20 daphnids divided into 4 groups with 5 animals per test vessel will be used. The immobilization of
the daphnids will be observed after 24 and 48 hours of exposure. Water parameters will be measured at test start and end of the study. Once per
month a reference item is tested to check the test system and test conditions.

GLP compliance:

yes (incl. QA statement)

Test material

Test material information

Constituent 1

Reference substance name:

3-hydroxypropiononitrile

EC Number:

203-704-8

EC Name:

3-hydroxypropiononitrile

Cas Number:

109-78-4

Molecular formula:

C3H5NO

IUPAC Name:

3-hydroxypropanenitrile

Details on test material:

- Supplier: Evonik Röhm GmbH, Darmstadt, Germany
- Purity: 99.1%
- Impurities: 0.46 % Ethylene glycol
- Lot/batch No.: 1038042701
- Expiration date of the lot/batch: 14 Nov. 2008
- Stability of test article dilution:
- Storage condition of test material: Room temperature, max. 30°C, protected from moisture and light

Sampling and analysis

Analytical monitoring:

yes

Test solutions

Vehicle:

yes

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: The stock solution (100 mg/L test item will be weighed out) will be freshly prepared with dilution water.

Dilution water: According to Directive 92/69/EEC Method C.2, Annex

Component Concentration [mg/L]
----------------------------------
KCL 5.76
NaHCO3 64.8
CaCl2 * 2 H2O 294
MgSO4 * 7 H2O 123
-----------------------------------
pH-value: 7.8 ±0.2

- Controls: Dilution water without test item tested under the same conditions as the test groups.
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): dilution water
- Concentration of vehicle in test medium (stock solution and final test solution): 100 mg/L

DOC - Analysis of test item Ethylöene cyanhydrin

DOC - Analysis of New (0h) and Old Media (48h)

Nominal
Test Item Concentration
[mg/L] DOC [mg/L]
0 h 48h Overall
-------------------------------------------------------------------------------------------------------------------------------------
Parallel determination Mean Parallel determination Mean Mean Corrected vs. Control
100 56.6 56.3 56.5 56.3 54.8 55.6 56.1 51.8
Control 4.3 4.3 4.3 4.3 4.3 4.3 4.3 --
-------------------------------------------------------------------------------------------------------------------------------------
vs. = versus

The measured DOC-values well met the expected range of the test concentrations (the theoretic carbon content of the test item is 50.2 %).

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Daphnia magna
- Strain: STRAUS (Clone 5)
- Source: Institut für Wasser-, Boden- und Lufthygiene (WaBoLu), Germany
- Age at study initiation (mean and range, SD): 2 - 24 h old
- Method of breeding: Breeder DR.U.NOACK-LABORATORIEN, Käthe-Paulus-Str. 1, D-31157 Sarstedt, Germany
Culture In 2-3 L glass vessels with approximately 1.8 L culture medium, at 20 ±2 °C, in an incubator,
16 h illumination, illumination strength max. 20 µE * m-2 * s-1.
Culture medium Elendt M4, according to ELENDT (1990), modified to a total hardness of 160 to 180 mg CaCO3/L.
Feeding At least 5 times per week ad libitum with a mix of unicellular green algae, e.g. Pseudokirchneriella
subcapitata, Desmodesmus subspicatus and Chlorella vulgaris, with an algae cell density of
> 106 cells/mL.
- Feeding during test: The daphnids will not be fed during the study.

ACCLIMATION
- Acclimation period: 2 h
- Acclimation conditions (same as test or not): At least 2 h in dilution water

Study design

Test type:

static

Limit test:

yes

Total exposure duration:

48 h

Test conditions

Test temperature:

19 - 20 °C

pH:

7.8 ± 0.2

Dissolved oxygen:

ca. 8 mg/L

Nominal and measured concentrations:

100 mg/L nominal;
measured test concentrations (DOC) correspond to theoretical DOC and are stable throughout the test (see Details on Test Solutions)

Details on test conditions:

TEST SYSTEM
- Test vessel: Glass beakers (4 cm ID x 7 cm H), 50 mL capacity
- No. of organisms per vessel: 5 animals per limit concentration and control
- No. of vessels per concentration (replicates): 4
- No. of vessels per control (replicates): 4

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Dilution water: According to Directive 92/69/EEC Method C.2, Annex

Component Concentration [mg/L]
----------------------------------
KCL 5.76
NaHCO3 64.8
CaCl2 * 2 H2O 294
MgSO4 * 7 H2O 123
-----------------------------------
pH-value: 7.8 +/-0.2

- Total organic carbon: 274 mg CaCO3/L
- Conductivity: 665 µS/cm
- Dissolved O2-concentration: 8.06 mg/L
- Intervals of water quality measurement: -


OTHER TEST CONDITIONS
- Adjustment of pH: -
- Photoperiod: 16/8 h light/dark cycle
- Light intensity: Diffuse light, illumination strength max. 20 µE*m-2 * s-1

Reference substance (positive control):

yes

Remarks:

Potassium dichromate p.a. (Sigma)

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 2

Duration:

48 h

Dose descriptor:

EC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 3

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 4

Duration:

48 h

Dose descriptor:

EC100

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 5

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Effect concentrations 6

Duration:

48 h

Dose descriptor:

LOEC

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Results with reference substance (positive control):

- Results with reference substance valid? yes

Results of the Reference Item potassium dichromate:

The percentage immobility for the reference item potassium dichromate was determined after 24 h. The EC50-value (24 h) and the 95 % confidence
interval (CI) were determined by sigmoidal dose-response regression to be:
EC50 : 1.64 mg/L (CI 1.47 - 1.82 mg/L)
The 24 h EC50-value for potassium dichromate was found to be within the prescribed concentration range of 1.0 - 2.5 mg/L which is in line with the quality criteria stated in AQS P 9/2 (05/1996) for daphnids clone 5 cultured in Elendt M4 medium. The EC50-value of the reference item is also within the recommended range of 0.6 - 2.1 mg/L according to OECD-Guideline 202.

Reported statistics and error estimates:

EC-values and statistical analyses for reference item

EC50-value will be only calculated for the reference item by sigmoidal dose-response regression. Calculation of the confidence intervals for EC50
will be carried out using standard procedures according to CLOPPER and PEARSON (1934).

Software
All data will be computer generated and rounded for presentation from the full derived data. Consequently if calculated manually based on the given
data minor variations may occur from these figures.
Calculations will be carried out using software:
- GraphPad Prism4, GRAPHPAD SOFTWARE, INC.
- Microsoft® Office Excel, MICROSOFT CORPORATION

Any other information on results incl. tables

Results of the Preliminary Test

The non GLP preliminary range finding test was conducted with three test item concentrations in the range of 10 to 1000 mg/L with a dilution factor of 10 under static conditions.

The percentage immobility was determined in all concentration levels and in the control after 24 and 48 hours of exposure. The immobilization rates are given in the table below.

Immobilization Rates of the Finding Test [%] (n = 20, divided into 2 replicates with 10 daphnids each)       

Nominal Test Item Concentration [mg/L]	IMMOBILIZATION [%]
	24 h			48 h
	Replicates			Replicates
	1	2	MV	1	2	MV
1000	0	0	0	0	0	0
100	0	0	0	0	0	0
10	0	0	0	0	0	0
Control	0	0	0	0	0	0

                              

MV= Mean value

Results of the Definitive Test

The tested limit concentration of 100 mg/L was visually clear i.e. completely dissolved throughout exposure. The percentage immobility was determined in the limit concentration and the control after 24 and 48 h.
The immobilization rates in % after 24 and 48 h of exposure are given in the tabe below. There was no biological effect, neither in the limit concentration nor in the control group.

Percentage of Daphnids Incapable of Swimming after 24 and 48 h of Exposure (n = 20, divided into 4 replicates with 5 daphnids each)

Nominal Test Item Concentration [mg/L]	IMMOBILIZATION [%]					48 h
	Replicates					Replicates
	1.	2.	3.	4.	MV	1.	2.	3.	4.	MV
100	0	0	0	0	0	0	0	0	0	0
Control	0	0	0	0	0	0	0	0	0	0

MV= Mean value

Results: EC-values, NOEC and LOEC (based on the nominal test item concentration)

	Test Duration [h]	Nominal Test Item Concentration [mg/L]	Confidence Interval p = 95% [mg/L]
EC50	24	> 100	not applicable
	48	> 100	not applicable
EC100	24	> 100	---
	48	> 100	---
NOEC	48	100	---
LOEC	48	> 100	---

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Conclusions:

At the limit concentration of 100 mg/L (nominal) of the test item Ethylene cyanohydrin no biological effect was observed.

Executive summary:

In the acute immobilization test with Daphnia magna (STRAUS) the effects of the limit concentration of 100 mg/L of the test item Ethylenecyanohydrin (batch number: 1038042701), was determined according to OECD 202 (2004). The limit test was conducted under static conditions over a period of 48 h from 2008-09-23 to 2008-09-25 at Dr. U. Noack-Laboratorien, D-31157 , Germany .

20 test organisms were exposed to the limit concentration and the control. The limit concentration and the control were analysed via DOC and GC-MS after 0 h and 48 h. Details of the DOC analytic are given in (Details on analytical methods).

All effect values are given based on the nominal concentration. Water quality parameters, pH-value and dissolved oxygen concentration, measured at 0 and 48 h, were determined to be within the acceptable limits. The validity criteria of the test guideline were fulfilled.

At the limit concentration of 100 mg/L (nominal) of the test item Ethylene cyanohydrin no biological effect was observed.