C14-16 (even numbered) and C16 (branched) saturated and unsaturated aliphatic hydrocarbons

Long-term toxicity to aquatic invertebrates

Administrative data

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

experimental study

Adequacy of study:

key study

Study period:

July - August 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This robust summary has a reliability rating of 2 because the study followed a standard guideline and was conducted without deviations that would invalidate the study. However, the study did not follow GLP guidelines.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.

Cross-reference

Reason / purpose for cross-reference:

read-across: supporting information

Reference

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

read-across from supporting substance (structural analogue or surrogate)

Adequacy of study:

key study

Study period:

July - August 1995

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: This robust summary has a reliability rating of 2 because the study followed a standard guideline and was conducted without deviations that would invalidate the study. However, the study did not follow GLP guidelines.

Justification for type of information:

A discussion and report on the read across strategy is given as an attachment in IUCLID Section 13.

Reason / purpose for cross-reference:

read-across source

Qualifier:

according to guideline

Guideline:

other: OECD guideline 202 section 2: Effects on biotic systems (1984)

Deviations:

yes

Remarks:

The deviation did not impact the scientific integrity of the study

GLP compliance:

no

Analytical monitoring:

yes

Details on sampling:

Samples of the control and the Water Accomodated Fractions (WAFs) prepared on each loading rate were taken on days 0, 2, 5, 7 and 14 from the stirring vessels and at termination (composite of replicates) for analysis.

Vehicle:

no

Details on test solutions:

The test solutions were prepared as WAFs (water accomodated fractions). Individual WAFs were prepared by adding the appropriate amount of the test substance to 3.5 L of dilution water in a 4 L size glass aspirator bottle. The mixtures were stirred for approximately 24 hours on a magnetic stirplate with a Teflon coated stirbar. Stirring was initiated using a vortex of <10% of the static liquid depth (1.5 cm). Each vesesl was stoppered with a foil covered neoprene stopper. All mixtures appeared clear during the stirring period.
After stirring, WAFs were removed through the outlet at the bottom of the vessel and added to test chambers. New WAF solutions were prepared for each 24-hour renewal.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: cultured in the Environmental Toxicology and Chemistry Laboratory of Exxon Biomedical Sciences, INC.
- Age of parental stock: 16 days
- Feeding during test
- Food type: Selenastrum capricornutum (algae)
- Amount: 2.32*10^5 cells/mL on days 0-6 and 1.56*10^5 cells/mL on days 7-21
- Frequency: daily

ACCLIMATION
- Acclimation period: no acclimation period (daphnia <= 24 hours old)

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES
Young were observed, counted and removed daily.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21

Remarks on exposure duration:

d

Post exposure observation period:

No post exposure period

Hardness:

No data

Test temperature:

Mean test temperature (water): 20°c (sd=0.05), continuously monitored by computer in test area.

pH:

6.6 TO 7.8

Dissolved oxygen:

5.2 TO 9.3 mg/L

Salinity:

Freshwater study

Nominal and measured concentrations:

Range finding test: Nominal concentrations (loading rates): 50, 10, 2.5, 1 and 0.5 mg/L. Samples of the control and the 50 mg/L loading were analyzed for the test substance by gas chromatography with flame ionization detection (GC-FID). The results of the analysis showed that the levels of the test susbtance were below the practical quantitation limit of 0.2 mg/L.
Definitive test: 0, 1, and 5 mg/L (nominal concentrations = loading rates)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed (flasks stoppered with foil covered neoprene stoppers on days 0-6; flasks covered with plexiglass sheets on days 7-21)
- Material, size, headspace, fill volume: 500 mL size glass erlenmeyer flasks, containing approximately 560mL of solution on days -6 (no head space) and 500 mL solution on days 7-21; test chambers rinsed with reverse osmosis water after renewals to remove food substrates
- Aeration: no aeration
- Renewal rate of test solution: renewal of the test solutions was performed on Monday, Wednesday and Friday by transferring the adult daphnids to new solutions via pipette.
- No. of organisms per vessel: range-finding test: 5; definitve test: 10
- No. of vessels per concentration (replicates): range-finding test: 2 (two trials); definitive test: 4
- No. of vessels per control (replicates): range-finding test: 2 (two trials); definitive test: 4
- Biomass loading rate: definitive test: 1 daphnid per approximately 50 to 56 mL of solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: BW3 supplemented with vitamin B12 and selenium
- Culture medium different from test medium: no
- Intervals of water quality measurement: The quality of the dilution water used in culture and testing is monitored at weekly, monthly, semi-annual and annual intervals.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h ightL/ 8 h Dark with gradual intensity conversion between periods
- Light intensity: approximately 630 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Adult mortality / immobilization
- Percent reduction in reproduction (based on cumulative young per adults)

RANGE-FINDING STUDY
- Test concentrations: 0, 0.5, 1.0, 2.5, 10, and 50 mg/L
- Loadings used for the definitive study: 0, 1, and 5 mg/L

Reference substance (positive control):

no

Duration:

21 d

Dose descriptor:

EL50

Effect conc.:

> 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

reproduction

Remarks:

cumulative average young per adult

Duration:

21 d

Dose descriptor:

EL50

Effect conc.:

> 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

immobilisation

Remarks on result:

other: (adults)

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

reproduction

Remarks:

cumulative average young per adult

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

immobilisation

Remarks on result:

other: (adults)

Details on results:

The first brood were observed in the control and each loading after 7 days. By day 14 a cumulative average of 20 young per female were produced in the control. A cumulative average of 54 young per female were produced in the control by day 21.
Other details on results in §Remarks on results including tables and figures.

Results with reference substance (positive control):

No reference susbtance

Reported statistics and error estimates:

The No Observed Effect Loading Rate (NOELR) was determined using Dunnett's procedure of SAS.

Table 1. Definitive test: Percent adult mortality / immobilization and percent reduction in reproduction (based on cumulative youngs per adult) for the 14- and 21-day period.

 

Loading rate (mg/L)	% Adult Mortality / Immobilization		% Reduction in reproduction
	14 days	21 days	14 days	21 days
Control	7.5	17.5	N/A	N/A
1.0	10	12	0	9
5.0	10	25	25	28

N/A - Not applicable, % reduction in reproduction as compared to the controls.

Validity criteria fulfilled:

yes

Remarks:

Dissolved oxygen fell below 60% sat. on day 12 in old solution. Since treatment was renewed, not believed to have effect on study. At day 21, 54 young per adult rather than 60, but believed sufficient young were produced for evaluation.

Conclusions:

The 21-day EL50s based on adult immobilization and reduction in reproduction were >5 mg/L. The 21-day No Observed Effect Loading Rates (NOELRs) based on adult immobilization and reduction in reproduction was 5 mg/L.

Executive summary:

The 21-day EL50s based on adult immobilization and reduction in reproduction were >5 mg/L loading. The 21-day No Observed Effect Loading Rates (NOELRs) based on adult immobilization and reduction in reproduction was 5 mg/L.

The study was performed to evaluate the toxicity of the water accomodated fraction (WAF) of the test substance to the daphnid, Daphnia magna, in a semi-static system for a 21-day period. The loading rates for this test were 1 mg/L and 5 mg/L. A control of laboratory dilution water was also tested. Individual WAFs were prepared by adding the appropriate amount of the test substance to 3.5 liters of BW3 in a 4 L size glass aspirator bottle. The mixtures were then stirred for approximately 24 hours on a magnetic stirplate with a Teflon coated stirbar. Stirring was initiated using a vortex of <10% of the static liquid depth (1.5 cm). Each vessel was stoppered to prevent loss of volatiles. After stirring, the WAF was removed from the outlet at the bottom of the vessel.

Four replicates chambers were prepared per treatment by completely filling the test chambers with WAF (no headspace). Samples were removed from each WAF and the control on day 0, 2, 5, 7, 14, and at termination (composite of the replicate test chambers) for chemical analysis by gas chromatography with flame ionization detection (GS-FID). The results of the analysis showed that the levels of the test substance were below the practical quantitation limit of 0.2 mg/L.

Data source

Reference

Reference Type:

study report

Title:

Unnamed

Year:

1996

Report date:

1996

Materials and methods

Test guideline

Qualifier:

according to guideline

Guideline:

other: OECD guideline 202 section 2: Effects on biotic systems (1984)

Deviations:

yes

Remarks:

The deviation did not impact the scientific integrity of the study

GLP compliance:

no

Test material

Test material information

Constituent 1

Reference substance name:

Hydrocarbons, C13-C16, isoalkanes, cyclics, <2% aromatics

Molecular formula:

None available - not a single isomer

IUPAC Name:

Hydrocarbons, C13-C16, isoalkanes, cyclics, <2% aromatics

Details on test material:

Hydrocarbons, C13-C16, containing isoalkanes and cyclics with less than 2% aromatics
- Substance type: UVCB
- Physical state: Clear liquid
- Analytical purity: 100% Commercial product
- Storage condition of test material: stored at room temperature

Sampling and analysis

Analytical monitoring:

yes

Details on sampling:

Samples of the control and the Water Accomodated Fractions (WAFs) prepared on each loading rate were taken on days 0, 2, 5, 7 and 14 from the stirring vessels and at termination (composite of replicates) for analysis.

Test solutions

Vehicle:

no

Details on test solutions:

The test solutions were prepared as WAFs (water accomodated fractions). Individual WAFs were prepared by adding the appropriate amount of the test substance to 3.5 L of dilution water in a 4 L size glass aspirator bottle. The mixtures were stirred for approximately 24 hours on a magnetic stirplate with a Teflon coated stirbar. Stirring was initiated using a vortex of <10% of the static liquid depth (1.5 cm). Each vesesl was stoppered with a foil covered neoprene stopper. All mixtures appeared clear during the stirring period.
After stirring, WAFs were removed through the outlet at the bottom of the vessel and added to test chambers. New WAF solutions were prepared for each 24-hour renewal.

Test organisms

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: cultured in the Environmental Toxicology and Chemistry Laboratory of Exxon Biomedical Sciences, INC.
- Age of parental stock: 16 days
- Feeding during test
- Food type: Selenastrum capricornutum (algae)
- Amount: 2.32*10^5 cells/mL on days 0-6 and 1.56*10^5 cells/mL on days 7-21
- Frequency: daily

ACCLIMATION
- Acclimation period: no acclimation period (daphnia <= 24 hours old)

METHOD FOR PREPARATION AND COLLECTION OF EARLY INSTARS OR OTHER LIFE STAGES
Young were observed, counted and removed daily.

Study design

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21

Remarks on exposure duration:

d

Post exposure observation period:

No post exposure period

Test conditions

Hardness:

No data

Test temperature:

Mean test temperature (water): 20°c (sd=0.05), continuously monitored by computer in test area.

pH:

6.6 TO 7.8

Dissolved oxygen:

5.2 TO 9.3 mg/L

Salinity:

Freshwater study

Nominal and measured concentrations:

Range finding test: Nominal concentrations (loading rates): 50, 10, 2.5, 1 and 0.5 mg/L. Samples of the control and the 50 mg/L loading were analyzed for the test substance by gas chromatography with flame ionization detection (GC-FID). The results of the analysis showed that the levels of the test susbtance were below the practical quantitation limit of 0.2 mg/L.
Definitive test: 0, 1, and 5 mg/L (nominal concentrations = loading rates)

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type: closed (flasks stoppered with foil covered neoprene stoppers on days 0-6; flasks covered with plexiglass sheets on days 7-21)
- Material, size, headspace, fill volume: 500 mL size glass erlenmeyer flasks, containing approximately 560mL of solution on days -6 (no head space) and 500 mL solution on days 7-21; test chambers rinsed with reverse osmosis water after renewals to remove food substrates
- Aeration: no aeration
- Renewal rate of test solution: renewal of the test solutions was performed on Monday, Wednesday and Friday by transferring the adult daphnids to new solutions via pipette.
- No. of organisms per vessel: range-finding test: 5; definitve test: 10
- No. of vessels per concentration (replicates): range-finding test: 2 (two trials); definitive test: 4
- No. of vessels per control (replicates): range-finding test: 2 (two trials); definitive test: 4
- Biomass loading rate: definitive test: 1 daphnid per approximately 50 to 56 mL of solution

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: BW3 supplemented with vitamin B12 and selenium
- Culture medium different from test medium: no
- Intervals of water quality measurement: The quality of the dilution water used in culture and testing is monitored at weekly, monthly, semi-annual and annual intervals.

OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod: 16h ightL/ 8 h Dark with gradual intensity conversion between periods
- Light intensity: approximately 630 Lux

EFFECT PARAMETERS MEASURED (with observation intervals if applicable):
- Adult mortality / immobilization
- Percent reduction in reproduction (based on cumulative young per adults)

RANGE-FINDING STUDY
- Test concentrations: 0, 0.5, 1.0, 2.5, 10, and 50 mg/L
- Loadings used for the definitive study: 0, 1, and 5 mg/L

Reference substance (positive control):

no

Results and discussion

Effect concentrations

Effect concentrations 1

Duration:

21 d

Dose descriptor:

EL50

Effect conc.:

> 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

reproduction

Remarks:

cumulative average young per adult

Effect concentrations 2

Duration:

21 d

Dose descriptor:

EL50

Effect conc.:

> 5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

immobilisation

Remarks on result:

other: (adults)

Effect concentrations 3

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

reproduction

Remarks:

cumulative average young per adult

Effect concentrations 4

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

5 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(loading rate)

Basis for effect:

immobilisation

Remarks on result:

other: (adults)

Details on results:

The first brood were observed in the control and each loading after 7 days. By day 14 a cumulative average of 20 young per female were produced in the control. A cumulative average of 54 young per female were produced in the control by day 21.
Other details on results in §Remarks on results including tables and figures.

Results with reference substance (positive control):

No reference susbtance

Reported statistics and error estimates:

The No Observed Effect Loading Rate (NOELR) was determined using Dunnett's procedure of SAS.

Any other information on results incl. tables

Table 1. Definitive test: Percent adult mortality / immobilization and percent reduction in reproduction (based on cumulative youngs per adult) for the 14- and 21-day period.

 

Loading rate (mg/L)	% Adult Mortality / Immobilization		% Reduction in reproduction
	14 days	21 days	14 days	21 days
Control	7.5	17.5	N/A	N/A
1.0	10	12	0	9
5.0	10	25	25	28

N/A - Not applicable, % reduction in reproduction as compared to the controls.

Applicant's summary and conclusion

Validity criteria fulfilled:

yes

Remarks:

Dissolved oxygen fell below 60% sat. on day 12 in old solution. Since treatment was renewed, not believed to have effect on study. At day 21, 54 young per adult rather than 60, but believed sufficient young were produced for evaluation.

Conclusions:

The 21-day EL50s based on adult immobilization and reduction in reproduction were >5 mg/L. The 21-day No Observed Effect Loading Rates (NOELRs) based on adult immobilization and reduction in reproduction was 5 mg/L.

Executive summary:

The 21-day EL50s based on adult immobilization and reduction in reproduction were >5 mg/L loading. The 21-day No Observed Effect Loading Rates (NOELRs) based on adult immobilization and reduction in reproduction was 5 mg/L.

The study was performed to evaluate the toxicity of the water accomodated fraction (WAF) of the test substance to the daphnid, Daphnia magna, in a semi-static system for a 21-day period. The loading rates for this test were 1 mg/L and 5 mg/L. A control of laboratory dilution water was also tested. Individual WAFs were prepared by adding the appropriate amount of the test substance to 3.5 liters of BW3 in a 4 L size glass aspirator bottle. The mixtures were then stirred for approximately 24 hours on a magnetic stirplate with a Teflon coated stirbar. Stirring was initiated using a vortex of <10% of the static liquid depth (1.5 cm). Each vessel was stoppered to prevent loss of volatiles. After stirring, the WAF was removed from the outlet at the bottom of the vessel.

Four replicates chambers were prepared per treatment by completely filling the test chambers with WAF (no headspace). Samples were removed from each WAF and the control on day 0, 2, 5, 7, 14, and at termination (composite of the replicate test chambers) for chemical analysis by gas chromatography with flame ionization detection (GS-FID). The results of the analysis showed that the levels of the test substance were below the practical quantitation limit of 0.2 mg/L.