Acetonitrile

Short-term toxicity to fish

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

1984

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

Guideline not specified

GLP compliance:

not specified

Analytical monitoring:

yes

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Age at study initiation (mean and range, SD): 26-13 days
- Length at study initiation (length definition, mean, range and SD): mean length 21.1 mm.
- Weight at study initiation (mean and range, SD): mean weight 0.165 g

Test type:

flow-through

Water media type:

freshwater

Hardness:

43.0 mg CaCO3/l

Test temperature:

26.1 degree C

pH:

pH 7.4

Dissolved oxygen:

6.1 mg/l.

Salinity:

alkalinity 46.0 mg CaCO3/l

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

1 640 mg/L

Executive summary:

Brooke, et. al. (1984) reported a 96-hour, flow-through LC50 value of 1640 mg/L for acetonitrile in Fathead minnow, (Pimephales promelas).

Reference 2

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

disregarded due to major methodological deficiencies

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: Basic data taken from Japanese paper.

Principles of method if other than guideline:

Information not available

GLP compliance:

not specified

Analytical monitoring:

not specified

Test organisms (species):

Oryzias latipes

Details on test organisms:

- Common name: Medaka, high-eyes

Test type:

semi-static

Water media type:

freshwater

Duration:

48 h

Dose descriptor:

other: TLm

Effect conc.:

730 mg/L

Executive summary:

Nishiuchi (1981) reported a 48-hour TLm value of 730 mg/L for acetonitrile in Medaka, high-eyes, (Oryzias Latipes).

Reference 3

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

The method used followed that of Doudoroff et al. 1951 (Aquire, 1994). The bibliographic details were not presented.

GLP compliance:

no

Analytical monitoring:

not specified

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill
- Length at study initiation (length definition, mean, range and SD): 3.8-5.1 cm
- Weight at study initiation (mean and range, SD):  2 g

Test type:

static

Water media type:

freshwater

Hardness:

20 mg CaCO3/l.

Test temperature:

25 degree C.

pH:

7.4

Dissolved oxygen:

8.0 mg/l.

Salinity:

Alkalinity 16 mg CaCO3/l, acidity 2.0 mg/l.

Duration:

96 h

Dose descriptor:

other: TLm

Effect conc.:

1 850 mg/L

Executive summary:

Henderson et al (1961) reported a 96 -hour, static LC50 value of 1850 mg/L for acetonitrile in Bluegill, (Lepomis macrochirus).

Reference 4

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

The method used followed that of Doudoroff et al. 1951 (Aquire, 1994). The bibliographic details were not presented.

GLP compliance:

no

Analytical monitoring:

not specified

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Length at study initiation (length definition, mean, range and SD): 5.1-6.4 cm in length
- Weight at study initiation (mean and range, SD): 1.5 g

Test type:

static

Water media type:

freshwater

Test temperature:

The test was conducted at 25 degree C.

Dissolved oxygen:

8.0 mg/l dissolved oxygen

Duration:

96 h

Dose descriptor:

other: TLm

Effect conc.:

1 000 mg/L

Executive summary:

Henderson et. al (1961) reported a 96-hour, static LC50 value of 1000 mg/L for acetonitrile in Fathead minnow, (Pimephales promelas).

Reference 5

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Principles of method if other than guideline:

The method used followed that of Doudoroff et al. 1951 (Aquire, 1994). The bibliographic details were not presented.

GLP compliance:

no

Analytical monitoring:

not specified

Test organisms (species):

Poecilia reticulata

Details on test organisms:

TEST ORGANISM
- Common name: Guppy
- Length at study initiation (length definition, mean, range and SD): 2.5 cm 
- Weight at study initiation (mean and range, SD): 0.1 g

Test type:

static

Water media type:

freshwater

Hardness:

20 mg/l

Test temperature:

25 degree C.

pH:

7.4

Dissolved oxygen:

8.0 mg/l

Salinity:

alkalinity 16 mg/l, acidity 2.0 mg/l.

Duration:

96 h

Dose descriptor:

other: TLm

Effect conc.:

1 650 mg/L

Executive summary:

Henderson et al (1961) reported a 96 -hour, static LC50 value of 1650 mg/L for acetonitrile in Guppy, (Poecilia reticulata).

Reference 6

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Principles of method if other than guideline:

The method evaluates the potential of a substance to cause acute aquatic effects in seven juvenile aquatic species simultaneously.

GLP compliance:

not specified

Analytical monitoring:

no

Test organisms (species):

Pimephales promelas

Details on test organisms:

TEST ORGANISM
- Common name: Fathead minnow
- Age at study initiation (mean and range, SD): Juvenile

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Hardness:

130 mg CaCO3/l.

Test temperature:

19-21 degree C.

pH:

6.5 - 8.5

Dissolved oxygen:

>= 40% of initial value.

Salinity:

alkalinity 46.0 mg CaCO3/l

Nominal and measured concentrations:

0.1, 1, 10, 100 mg/L

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Executive summary:

Ewell et. al. (1996) reported a 96-hour LC50 value of >100 mg/L for acetonitrile in Fathead minnow, (Pimephales promelas).

Reference 7

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

documentation insufficient for assessment

Qualifier:

according to guideline

Guideline:

other: Deutsche Einheitsverfahen zur Wasser-, Abwasser- und Schlamm Untersuchung. L.15: Fischtest, 1976

Principles of method if other than guideline:

Deutsche Einheitsverfahren zur Wasser-, Abwasser- und Schlamm Untersuchung. L.15: Fischtest. Vom Wasser 46: 291-295, 1976.

GLP compliance:

not specified

Analytical monitoring:

not specified

Test organisms (species):

Leuciscus idus melanotus

Details on test organisms:

- Common name: Golden orfe

Test type:

static

Water media type:

freshwater

Duration:

48 h

Dose descriptor:

LC0

Effect conc.:

3 900 - 4 330 mg/L

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

5 850 - 7 050 mg/L

Duration:

48 h

Dose descriptor:

LC100

Effect conc.:

6 240 - 8 670 mg/L

Executive summary:

Juhnke and Luedemann (1978) reported 48-hour, static LC50 values for acetonitrile in Golden orfe, (Leuciscus idus melanotus) from two different laboratories of 5850 mg/L and 7050 mg/L.

Reference 8

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

23 October 1995 - 29 March 1996

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

OECD Guideline 203 (Fish, Acute Toxicity Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

- Physical state: colourless and transparent liquid
- Analytical purity: 100.0% (GC)
- Lot/batch No.: ESH3487
- Stability under test conditions: Stable
- Storage condition of test material: In the refrigerator

Analytical monitoring:

yes

Details on sampling:

- Sampling: 0, 24h
- Sampling volume: 2 mL

Vehicle:

no

Details on test solutions:

A test solution used in this study was prepared by diluting the test substance with the dechlorinated tap water.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Orange killifish
- Source: Niikura fish farm
- Length at study initiation: 2.1 cm (1.9 - 2.4 cm), n=10
- Weight at study initiation: 0.18 g (0.12 - 0.25 g), n=10
- Feeding during test: none


ACCLIMATION
- Acclimation period: 45 days
- Acclimation conditions: Same as test
- Type and amount of food: TetraMin

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

96 h

Test temperature:

23.3 - 24.0℃

pH:

7.4 - 8.0

Dissolved oxygen:

6.5 - 8.5 mg/L

Nominal and measured concentrations:

Nominal concentration: 100 mg/L (and control)
Mean (geometric mean) measured concentration: 102 mg/L (and control)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material: glass beaker
- Size: 5.0L
- Fill volume: 5.0 L
- Aeration: No
- Renewal rate of test solution: 24h
- No. of organisms per vessel: 10
- No. of vessels per concentration (replicates): 1
- No. of vessels per control (replicates): 1


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: dechlorinated tap water
- Hardness of dilution water: 50 mg/L (as CaCO3)
- pH of dilution water: 7.9


OTHER TEST CONDITIONS
- Photoperiod: 16h light/8h dark



TEST CONCENTRATIONS
- Range finding study
- Test concentrations: 0, 1000 mg/L
- Results used to determine the conditions for the definitive study: 0% mortality at 1000 mg/L within the period of 96h test

Reference substance (positive control):

yes

Remarks:

Copper(II) sulfate pentahydrate

Duration:

96 h

Dose descriptor:

LC50

Effect conc.:

> 100 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality (fish)

Details on results:

- Behavioural abnormalities: no observation
- Mortality of control: 0

Results with reference substance (positive control):

- 96h LC50: 0.88 mg/L

Conclusions:

The 96 -hr LC50 for acetonitrile in fish (Oryzias latipes) is concluded to be >100 mg/L (nominal conc.).

Executive summary:

In a guideline (OECD 203) study under GLP conditions, MCSI of Japan (1996) reported the 96 -hr LC50 for acetonitrile in fish (Oryzias latipes) to be >100 mg/L (nominal conc.).

Reference 9

Endpoint:

short-term toxicity to fish

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

study well documented, meets generally accepted scientific principles, acceptable for assessment

Qualifier:

according to guideline

Guideline:

other: Procedure of Japan Industrial Standards (Japanese Industrial Standards Committee, 1971).

GLP compliance:

not specified

Analytical monitoring:

no

Details on test solutions:

Chemical substances were dissolved in water and neutralized with 0.01 N NaOH or HCI, if necessary.

Test organisms (species):

Oryzias latipes

Details on test organisms:

TEST ORGANISM
- Common name: Medaka, high-eyes
- Length at study initiation: 2 cm
- Weight at study initiation: 0.2 g

ACCLIMATION
- Acclimation period: 10 days in tap water

Test type:

static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

48 h

Test temperature:

25 degree C.

Nominal and measured concentrations:

1,000 mg/liter

Details on test conditions:

Ten fish of Himedaka per one trial were kept in 2 liter of deionized water at 25 degrees C and, after 24 or 48 hours, lethal concentration of 50% fish was determined.

TEST SYSTEM
Himedaka
- Test vessel: 1 liter flask
- Type (delete if not applicable): open / closed
- Material, size, headspace, fill volume: 50 mg/liter
- No. of organisms per vessel: 10 Himedaka
- Other: Electrode sensor of dissolved oxygen meter was inserted into the neck of the flask and the flask was kept at 25degreeC and occasionally mixed. Dissolved oxygen content in the flask was checked periodically.

Duration:

48 h

Dose descriptor:

other: TLm

Effect conc.:

>= 1 000 mg/L

Nominal / measured:

nominal

Sublethal observations / clinical signs:

TLm values and partition coefficient of acetonitrile

Substance	TLm (mg/liter)a	TLm (mg/liter)a	Partition coefficient between n -octanol and water
	(24 hr)	(48 hr)
Acetonitrile	1,000	1, 000	0.29

a) Ten fish of Himedaka were used in 2 liter of the solution.

Time course of accumulated dyes at gill and intestine of fish

Dye	Time(min)	Amount of accumulated dye (u/g)Gill	Amount of accumulated dye (u/g) Intestine	Number of fish died
Rose Bengale	30	1.0	0	0/10
	60	5.4	0	2/10
	180	73.5	0	5/10
Methylene Blue	30	2.1	0	1/10
	60	8.6	0	3/10
	180	105.8	0	7/10

Note. 1) Fish (carp) was exposed to a solution of dye (1,000 mg/liter). 2) Dye accumulated to organ was eluted and determined by spectrophotometry.

Number of red corpuscles of fish kept under three different conditions

Exposed solution	Number of red corpuscle (x 10^4/ml)
A. Water	80 (Fish alive)
B. Rose Bengale (1,000 mg/liter)	130 (Fish died)
C. Water added with dry ice to evolve dissolved oxygen	120 (Fish died)

Note. Fish (carp) was exposed to each solution, and the number of red corpuscle in fish blood was determined by Coulter counter after 24 hours in case of A and immediately after fish had died in case of B and C.

Executive summary:

Tonagai et. al. (1982) reported a 48-hour, static median tolerance limit (TLm) value of 1000 mg/L for acetonitrile in Medaka, high-eyes, (Oryzias latipes).

Description of key information

The acute toxicity of acetonitrile to fish has been studied in several freshwater species.  Reported LC50 values range from 730 mg/L to 7050 mg/L.  Brooke et al (1984) reported the only flow-through study with a 96-hour LC50 value of 1640 mg/L in Pimephales promelas (Fathead minnow).  The lowest acute toxicity value reported for fish is a 48-hour TLm value of 730 mg/L reported by Nishiuchi in Oryzias latipes (Medaka, high-eyes), however given that it was not possible to obtain full text details from this paper, this data is disregarded.  Tonogai et al (1982) reported an 48-hour LC50 value of >1000 mg/L in the same species, which is consistent with findings in other fish species tested.  The reason for the low value in the Nishiuchi study is not clear.

Key value for chemical safety assessment

Fresh water fish

Fresh water fish

Effect concentration:

1 640 mg/L

Additional information

The acute toxicity of acetonitrile to fish has been studied in several freshwater species. Brooke et al, 1984 conducted exposure using flow-through apparatus; all other studies have used a static design. The fish toxicity data for acetonitrile were reviewed in the EU Risk Assessment Report (2000). A summary of the available data is shown in the table below. A single additional study has been obtained since the EU RAR. In a guideline (OECD 203) limit study under GLP conditions, MCSI of Japan (1996) reported the 96 -hr LC50 for acetonitrile in fish (Oryzias latipes) to be >100 mg/L (nominal conc.).

The acetonitrile data base includes values for Pimephales promelas, Lepomis macrochirus, Leuciscus idus melanotus, Poecilla reticulata and Oryzias latipes. The lowest value is the 48 h TLm, 730 mg/l, reported in IUCLID as toxicity for Oryzias latipes while all 48-96 h LC₅₀ values are in the g/L range. Information on additional fish species can be obtained from other sources. The WHO report (1993) includes 14 acute LC₅₀ values on seven fish species. Significant values are 24-48 h LC₅₀ values higher than 1g/L for Oryzias latipes, and two 48 h LC₅₀s, 730 and 880 mg/L on Cyprinus carpio and Ctenopharyngodon idellus respectively. The data set includes flow through and static test as well as different water pH and hardness conditions. Relevant differences on acetonitrile toxicity regarding water quality conditions were not observed.

The lowest acute toxicity value reported for fish is 730 mg/l as reported by Nishiuchi, although discrepancies between the fish species showing this toxicity appear when comparing IUCLID 2000 and the other reviews. The original paper has been submitted but it is in Japanese; however, the title and the scientific names also appear in English and indicate that the toxicity data are for the carp, Cyprinus carpio, and therefore is reported as toxicity for carp in the table below. In any case, the fish species is not relevant for this assessment and the value will be used as the lowest acute fish toxicity value.

Toxicity of Acetonitrile to Fish (EU Risk Assessment Report, 2000)

SPECIES	TEST TYPE	COMMENTS	DURATION	TOXICITY END POINT(mg/l)	REFERENCE
Pimephales promelas(fresh water)	Static nominal concentration	Used as additional information	96h	LC50=1000-1150	Henderson et al., (1961) (cited in UK and IPCS reports)
Pimephales promelas(fresh water)	Static nominal concentration	Valid	96h	LC50> 100	Ewell et al., (1986)
Lepomis macrochirus(fresh water)	Static nominal concentration	Used as additional information	24-46h	LC50= 1850	Henderson et al., (1961) (cited in UK and IPCS reports)
Ctenopharyngodon idellus	Static nominal concentration	Used as additional information	24-48h	LC50= 1950 LC50= 880	Chen, (1981) (cited in IPCS)
Poecilia reticulata(fresh water)	Static nominal concentration	Used as additional information	24-96h	LC50= 1650	Henderson et al, (1961) (cited in UK and IPCS reports)
Oryzias latipes(fresh water)	Static nominal concentration	Used as additional information	24-48h	LC50> 1000	Tonogai et al., (1982) (cited in UK and IPCS reports)
Cyprinus carpio(fresh water)	--	Valid	48h	LC50= 730	Nishiuchi (1981)
Pimephales promeles(fresh water)	flow through measured concentration	Valid	96h	LC50=1640	Brooke et al, 1984 Centre for Lake Superior Environmental studies
Leciscus idus melanotus(fresh water)	Static	Used as additional information	48h	LC50= 5850-7050	Juhnkeand Ludemann (1978) (cited in UK and IPCS report)