3,5,5-trimethylcyclohex-2-enone

Short-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

other information

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with national standard methods with acceptable restrictions

Principles of method if other than guideline:

Method: other: Acute Toxicity Test

GLP compliance:

not specified

Analytical monitoring:

not specified

Test organisms (species):

Artemia salina

Details on test organisms:

TEST ORGANISMS
- Source/supplier: dried eggs from Carolina Biological Supply Co.,  Burlington, N.C.
- Breeding method: aeration in synthetic seawater until hatching was  completed; settling out of unhatched eggs, concentration of shrimps in  
beam of light and transport to separate container
- Control group: synthetic seawater
- Age: 48 hours

Test type:

static

Water media type:

saltwater

Limit test:

no

Total exposure duration:

24 h

Test temperature:

24.5 °C

Nominal and measured concentrations:

0, 100, 180, 320, 560, 1000 mg/l (selection based on screening test)

Details on test conditions:

DILUTION WATER
- Synthetic seawater: 557.37 g NaCl, 27.20 g CaSO4, 63.36 g MgSO4 x 7 H2O,   168.30 g MgCl2, 15.84 g KCl, 3.14 g MgBr2 x 6 H2O,   all dissolved in 20 l of distilled water in this order

TEST SYSTEM
- Concentrations: 100, 180, 320, 560, 1000 mg/l   (selection based on screening test)
- Renewal of test solution: no
- Exposure vessel type: 150 ml wide-mouth bottles (test volume of 100 ml)
- Number of replicates, individuals per replicate: 1; 30-50 (up to 48 h old)

TEST PARAMETER
- immobility (no movement of the phyllopodia = appendages)

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

430 mg/L

Nominal / measured:

nominal

Basis for effect:

mobility

Remarks on result:

other: Endpoint used was TLm (median tolerance limit)

Details on results:

A straight line was obtained by plottting the percent survival versus the test dosage concentration (logarithmic scale). The concentration at which this line crosses 50 percent survival is reported as the TLm (median Tolerance Limit).

Executive summary:

The saltwater shrimp Artemia salina was exposed to 5 different concentrations of the substance for a period of 24 h. The following endpoint was derived

EC50 (referred to as TLm): 430 mg/L

The test was conducted according to standard methods and was assessed as "reliable with restrictions".

Reference 2

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

supporting study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Test procedure in accordance with national standard methods with acceptable restrictions

Qualifier:

according to guideline

Guideline:

other: DIN 38412, part 11

Deviations:

not specified

Principles of method if other than guideline:

Method: other: DIN 38412 part 11

GLP compliance:

no

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Concentration: 2 g/l, no solvent or vehicle

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISMS
- Strain: Daphnia magna, Hüls
- Source/supplier: Hüls AG (inhouse)
- Breeding method: in 1 l jars with dechlorinated drinking water, water renewal every 2-3 days, isolation of juveniles for further breeding after 4 weeks
- Feeding: Chlorella vulgaris, as much as consumed
- Pretreatment: Filtration of adults 24 h prior to testing

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

24 h

Test temperature:

20 +/- 1 °C

Nominal and measured concentrations:

control, 60; 90; 120; 180; 250; 350; 500; 700; 1000 mg/l (nominal concentrations)

Details on test conditions:

TEST SYSTEM
- Exposure vessel type: 25 ml graduated cylinder
- Number of replicates, individuals per replicate: 4 replicates with 5 individuals each
- Age at study start: < 24 hours
- Feeding during test: no
- Control group: 2 reference substance controls, one blank

TEST PARAMETER
- immobilisation

DILUTION WATER
- Source: Synthetic:   
- CaCl2 x 2 H2O: 294 mg/l   
- MgSO4 x 7 H2O: 123 mg/l   
- NaHCO3: 63 mg/l   
- KCl: 5.5 mg/l
- Ca/Mg ratio: 4:1
- Na/K ratio: 10:1

Reference substance (positive control):

yes

Remarks:

 potassium dichromate, CAS RN 7778-50-9

Duration:

24 h

Dose descriptor:

EC0

Effect conc.:

90 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Duration:

24 h

Dose descriptor:

EC50

Effect conc.:

254 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Remarks on result:

other: CL 221-293

Duration:

24 h

Dose descriptor:

EC100

Effect conc.:

500 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mobility

Details on results:

EC50 values were calculated using probit analysis according to Cavalli-Sforza 1972

Results with reference substance (positive control):

RESULTS: TEST WITH REFERENCE SUBSTANCE
- Concentrations: 0.9 / 1.9 mg/l
- Results: 5 / 95 % immobilisation

Reported statistics and error estimates:

95 % confidence interval of LC50: 221-293 mg/l 

- Concentration / response table: 
  60 / 90 mg/l: 0 % immobile
  120 / 180 mg/l: 15 % immobile
  250 mg/l: 50 % immobile
  350 mg/l: 65 % immobile
  500 / 700 / 1000 mg/l: 100 % immobile

Conclusions:

The EC50 value was determined as 254 mg/L indicating that the test item does not pose a significant hazard to aquatic invertebrates.

Executive summary:

The acute effects on Daphnia magna were investigated in a static 24h test in accordance with the national guideline DIN 38412, part 11. Nine concentrations ranging from 60 to 1000 mg/L were tested. The following endpoints were derived

EC50 (24h): 254 mg/L;       95% CL: 221- 293 mg/L

The study was assessed as "reliable with restrictions".

Reference 3

Endpoint:

short-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Study in accordance with generally accepted scientific standards and described in sufficient detail

Qualifier:

according to guideline

Guideline:

other: Methods for acute toxicity tests with fish, macroinvertebrates, and amphibians, US EPA (1975)

Deviations:

not specified

GLP compliance:

not specified

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

STOCK AND TEST SOLUTION AND THEIR PREPARATION
- Vehicle, solvent: not reported, probably none
- A stock solution in distilled water was prepared
- Other procedures: Preparation depended on solubility. 
- Procedures are  reported but not assigned to substances.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISMS
- Source/supplier: EG&G Bionomics inhouse
- Age: < 24 hours
- Dilution water: deionized reconstituted well water
- pH of dilution water was adjusted to pH 8.0 +/- 0.2 according to a.m. guideline

Test type:

static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

48 h

Hardness:

173 +/- 13 mg/L as CaCO3

Test temperature:

22 +/- 1 °C

pH:

8.0 +/- 0.2

Dissolved oxygen:

> 60 % at study initiation
- ranged from 6.5-9.1 mg/l for all tests

Details on test conditions:

TEST SYSTEM
- Concentrations: 5 to 8, values not reported
- Renewal of test solution: no
- Exposure vessel type: 250 ml beaker
- Number of replicates, individuals per replicate: 3, 5
- when appropriate, a solvent control was used

Duration:

48 h

Dose descriptor:

NOEC

Effect conc.:

15 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Duration:

48 h

Dose descriptor:

LC50

Effect conc.:

120 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: CL 72-170

Duration:

24 h

Dose descriptor:

LC50

Effect conc.:

430 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: CL 360-500

Details on results:

The LC50's and 95% confidence limits were calculated using a moving average angle method or by Probit analysis after converting the nominal
concentrations to logarithms

Results with reference substance (positive control):

- no data

Validity criteria fulfilled:

not specified

Conclusions:

The LC50 value (48 hours) was calculated to be 120 mg/L indicating that the test substance does not pose a significant hazard to aquatic invertebrates.

Executive summary:

The acute effects of the test item on Daphnia magna were investigated in a static 48h test in accordance with general accepted scientific standards. The following endpoints were derived

LC50 (48h): 120 mg/L; 95% CL: 72 -170 mg/L; NOEC: 15 mg/L

The study was assessed as "reliable with restrictions".

Description of key information

The acute toxicity of the substance to Daphnia magna was determined in a static 48-h test conducted according to a U.S. EPA guideline (LeBlanc, 1980). A second acute toxicity study (24 h) was conducted by Hüls AG, 1996 according to DIN 38412 which is similar to OECD 202. Both studies were assessed as "reliable with restrictions".

Key value for chemical safety assessment

Fresh water invertebrates

Fresh water invertebrates

Effect concentration:

120 mg/L

Additional information

After 24 h of exposure, the LC50 was calculated to be 430 mg/l, while after a period of 48 h a LC50 of 120 mg/l and a NOEC of 15 mg/l were obtained (LeBlanc, 1980). Based on immobilization, the nominal EC50 of the study conducted by Huels AG, 1996 was reported to be 254 mg/l. The lowest available 48 -h LC50 was found to be 120 mg/l (LeBlanc, 1980) and will be considered as the relevant endpoint for the short-term toxicity of the test item to aquatic invertebrates.

For the saltwater shrimp Artemia salina an EC50 (referred to as TLm (median tolerance limit)) of 430 mg/L for an exposure period over 24 h was derived.