Hydrocarbons, C4, 1,3-butadiene-free, polymd., triisobutylene fraction, hydrogenated

Long-term toxicity to aquatic invertebrates

Link to relevant study record(s)

Reference

Reference 1

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:

24 Jun - 21 Nov 2016

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

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: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

GLP compliance:

yes (incl. QA statement)

Specific details on test material used for the study:

SOURCE OF TEST MATERIAL
- Source and lot/batch No.of test material: 0013861306
- Expiration date of the lot/batch: 15 Dec 2016
-Purity: 98.6%

Analytical monitoring:

no

Vehicle:

no

Details on test solutions:

PREPARATION AND APPLICATION OF TEST SOLUTION
- Method: The test substance is a mixture that is poorly soluble in water. Therefore the test solutions were prepared following general guidance provided in OECD 23 [Ref. 8] in order to achieve a water accommodated fraction (WAF) of the test substance using a liquid-liquid saturator technique. Each test solution was prepared separately by directly adding test substance to test medium. Glass aspirator bottles (2-L) with a bottom side-outlet attached to a stopcock were filled with 2-L test media. The necessary volume of the test substance was calculated from the density. Prior to use the test substance was homogenized by shaking the container. Then the test substance was pipetted carefully on the water surface. The bottle was closed and the solution was stirred on a magnetic plate for approximately 1 day. The stirring was slow enough (approx. 100 rpm) so that no vortex formed, preventing foaming or emulsification. After stirring, slight streaks were observed on the water surface in test concentrations ≥2.3 mg/L. The lower test concentrations appeared colorless and clear. The required volume of aqueous fraction (test solution) was drawn off for testing via the bottom of each bottle. All test solutions were visibly colorless and clear throughout each renewal period. The control medium is treated in the same way.

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: water flea
- Strain/clone: 1820
- Justification for species other than prescribed by test guideline:
- Source: supplied by the Institut National de Recherche Chimique Appliquée, France, in 1978. From this date on this clone was cultured and bred continuously in the Ecotoxicology Laboratory of Experimental Toxicology and Ecology, BASF SE, Ludwigshafen, Germany.
- Age of parental stock (mean and range, SD): <24 h
- Feeding during test:
- Food type: live green algae Desmodesmus subspicatus
- Amount: The daphnids were fed a defined volume (≤153 μL) of the
concentrate (1310 - 1440 mg TOC/L) to reach the following amount of food: 0-3d: 0.1 mg TOC/L; 4-5d: 0.12 mg TOC/L; 6-7d: 0.14 mg TOC/L; 8-21d: 0.2 mg TOC/L
- Frequency: daily

ACCLIMATION
- Acclimation period: none
- Acclimation conditions (same as test or not): same
- Type and amount of food: same as during test
- Health during acclimation (any mortality observed): Only young from healthy cultures without signs of stress are used for testing.

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

21 d

Hardness:

2.41 - 2.51 mmol/L

Test temperature:

20 °C

pH:

7.8 - 8.4

Dissolved oxygen:

8.2 - 9.0 mg/L

Nominal and measured concentrations:

Nominal: 0.52, 1.1, 2.3, 4.9 and 10.2 mg/L as loading rate WAF

Details on test conditions:

TEST SYSTEM
- Test vessel: 50 mL glass vessels containing 50 mL test solution
- Aeration: none
- Renewal rate of test solution (frequency/flow rate): daily renewal
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates): 10

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: synthetic freshwater (Elendt M4)
- Total organic carbon: < 2 mg/L
- Alkalinity: 0.8 1.0 mmol/L
- Ca/mg ratio: 4:1
- Conductivity: 550 - 650 µS/cm
- Culture medium different from test medium: no
- Intervals of water quality measurement: Throughout the test, the appearance of the test solutions and dissolution behavior of the test substance was observed and recorded daily. The chemical and physical parameters of the test medium (total hardness, acid capacity, pH, conductivity and total organic carbon) were determined after aeration and prior to use in the test and were within acceptable ranges. Dissolved oxygen, pH and temperature were measured in the freshly prepared test solution and in the old test solution in replicate 1 of each test group for one interval per week. In addition, temperature was measured continuously during the whole exposure period in a separate vessel filled with water proximal to the test vessels. Hardness was measured in the freshly prepared test solution in an additional replicate of each concentration for one interval per week and in the old test solution from the combined replicates of each test group for one interval per week.

OTHER TEST CONDITIONS
- Photoperiod: 16h light/ 8h dark
- Light intensity: 699–756 lux at a wave length of 400–750 nm. The light intensity will not exceed 1000–1500 lux for cool white light (equivalent to 15–20 μE•m-2•s-1 according to the test guideline)

EFFECT PARAMETERS MEASURED: Parent mortality, abnormal effects, and numbers of live and dead offspring were assessed every day throughout the experiment.

- Range finding study: A preliminary range finding test was performed (experimental conduct in accordance with GLP but without a GLP status), with exposure over 17 days and daily solution renewal, at the following nominal concentrations: 0 (control), 1, 3, 10 mg/L. No significant effects were observed at 1 and 3 mg/L. At 10 mg/L there was a small, but not significant, reduction in offspring observed.

Reference substance (positive control):

yes

Remarks:

sodium chloride

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

> 10.2 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Remarks:

(WAF)

Basis for effect:

reproduction

Details on results:

No additional adverse effects or abnormal behavior were observed in any of the other test groups.

Results with reference substance (positive control):

The EC50(48 h) of the reference substance sodium chloride (NaCl) was 5.36 g/L (experiment date: 14 Jun 2016, project number: 50E0789/12E060). This result is within the range of 3.88–7.22 g/L, which represents ±2 standard deviations from the published EC50(48 h) of 5.55 g/L [Ref. 6, 7] and indicates that the culture of Daphnia magna used in this study is responding normally to toxic stress.

There was no statistically significant effect on parent mortality (Fisher’s exact test) and reproduction per surviving and introduced parent (Wilcoxon test with Bonferroni-Holm adjustment) in any of the test groups.

Three mortalities were observed in the highest test group 5 (10.2 mg/L), two mortalities were observed in test group 4 (4.9 mg/L), one mortality each was observed in groups 3 (2.3 mg/L) and in the control group, although none of these were statistically significant according to Fisher’s test. In order to determine if these mortalities were inadvertent or substance related, the data were evaluated for a concentration response pattern using a step-down Cochran- Armitage trend test as indicated in the OECD 211 test guideline [Ref. 2]. The mortality data with test group 5 was significant; however, the data with test group ≤4 did not follow a concentration response pattern. Therefore the mortalities in group 3, group 4 and the control group is considered inadvertent. The inadvertent mortalities were excluded from the evaluation of reproduction per introduced parent, as recommended in the test guideline.

Reproduction per surviving and introduced parent was not significantly affected in anyt reatment group according to the Wilcoxon test. Although the reproduction per introduced parent in test group 5 is much less than the control, the standard deviation was very high due to three replicates that did not produce any offspring, while all other replicates produced offspring within the range of the control replicates.

Statistically significant effects on %immobile young were observed at ≥4.9 mg/L, however there was no effect on reproduction as living young in these test groups so the slight effect on immobile young is not considered toxicologically significant. On study day 17 a daphnid appeared to be pale and small in test group 2 (1.1 mg/L) in comparison to the animals in the control group. This appearance was transient as it was not observed anymore for the remainder of the test.

Validity criteria fulfilled:

yes

Conclusions:

The test substance is not harmful to aquatic invertebrates based on chronic exposure.

Executive summary:

The study was performed according to GLP and OECD guideline 211 using Daphnia magna as test organism (BASF 2017). Loading rate WAFS between 0.52 and 10.2 mg/L were tested. There was no statistically significant effect on parent mortality and reproduction per surviving and introduced parent in any of the test groups. Thus, the 21d-NOELr is determined to be >= 10.2 mg/L based on nominal loading rate.

Reference 2

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:

2001

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

other: This robust summary has a reliability rating of 1 because the study followed a standard guideline, followed GLP, and was conducted without deviations that would invalidate the study.

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: supporting information

Qualifier:

according to guideline

Guideline:

OECD Guideline 211 (Daphnia magna Reproduction Test)

Deviations:

no

GLP compliance:

yes

Analytical monitoring:

no

Details on sampling:

No analytical measurements were taken. The low solubility of the test substance at the loading level tested was expected to result in exposure levels below the analytical detection limit for Total Organic Carbon (TOC) or Dissolved Organic Carbon (DOC).

Vehicle:

no

Details on test solutions:

Individual treatment solutions were prepared by adding the test substance, via syringe, to 4 L of laboratory blend water in glass aspirator bottles that were sealed with stoppers to minimize volatile loss of test material. The stock solutions were mixed for 24 hours. Mixing was performed using a magnetic stir plate and Teflon stir bar. After mixing, the solutions were allowed to settle for one hour at test temperature. The Water Accommodated Fraction (WAF) was then removed through the outlet at the bottom of the mixing vessel and added to the test chambers.

Test organisms (species):

Daphnia magna

Details on test organisms:

The Daphnia were cultured in-house, were <24 hours old, and were from 17-day old parents. Test organisms were fed daily when solutions were renewed with Pseudokirchneriella subcapitata to provide approximately 4.7 x 10E5 cells/mL. The daphnia were fed a carbon equivalent of approximately 0.5 mg C per adult per day which is purposely higher than the guideline to achieve an acceptable level of neonate production.

Test type:

semi-static

Water media type:

freshwater

Limit test:

yes

Total exposure duration:

21 d

Post exposure observation period:

None

Hardness:

The hardness of the dilution water during the study was 114-120 mg/L as CaCO3.

Test temperature:

Mean test temperature was 20.6 degrees C (S.D. = 0.3) for test substance and control systems.

pH:

7.0 to 7.9 for test substance and control systems.

Dissolved oxygen:

7.3 to 8.7 mg/L for test substance and control systems.

Nominal and measured concentrations:

Nominal loading rates were control and 1 mg/L. Analytical measurements were not taken.

Details on test conditions:

The test chambers were 55 mL capacity glass Erlenmeyer flasks with ground glass stoppers (no headspace). One daphnid was added to each of 10 replicates. The test was performed using a static daily renewal of exposure solutions. Observations for abnormal or immobilized daphnids and neonates were made on each replicate at approximately 24-hour intervals. Diurnal light: approximately 17 hours light and 7 hours dark. Water Quality was measured twice per week.

Reference substance (positive control):

no

Key result

Duration:

21 d

Dose descriptor:

NOELR

Effect conc.:

1 mg/L

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks:

The test substance also showed no effects based on growth.

Details on results:

The following are the endpoint results for the control and test solution:
Adult Neonates
Group Immobilization per
(%) Adult
Control 20 61
1 mg/L 10 47

The control daphnids released their first brood between days 7 and 9. The coefficient of variation (cv) for control fecundity was 33%. The test was considered acceptable as control organisms immobilization did not exceed 20% and mean reproduction in the controls exceeded 60 young per adult.

Results with reference substance (positive control):

None used

Reported statistics and error estimates:

Statistical analyses were conducted using the T-test for reproduction from ToxStat 3.4 (West Inc. 1994).

Validity criteria fulfilled:

yes

Conclusions:

The test substance showed no effects on survival, reproduction, or length at a nominal loading of 1 mg/L. Therefore, the 21-Day No Observed Effect Loading Rate (NOELR) = 1.0 mg/L for these endpoints based upon nominal loading levels.

Executive summary:

The test substance showed no effects on survival, reproduction, or length at a nominal loading of 1 mg/L. Therefore, the 21-Day No Observed Effect Loading Rate (NOELR) = 1.0 mg/L for these endpoints based upon nominal loading level.

Reference 3

Endpoint:

long-term toxicity to aquatic invertebrates

Type of information:

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

Adequacy of study:

weight of evidence

Study period:

from 2001-07-24 to 2001-08-1

Reliability:

4 (not assignable)

Rationale for reliability incl. deficiencies:

other: see 'Remark'

Remarks:

Well documented study. However, information on the substance is lacking, particularly the density of the susbtance would have been necessary to be able to calculate the end-point values as concentrations (mg/L for instance) instead of % WAF as it is in the study report. MSDS needed.

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: supporting information

Qualifier:

according to guideline

Guideline:

other: EPA 1002.0: Ceriodaphnia survival and reproduction test

Deviations:

no

Principles of method if other than guideline:

Ceriodaphnia dubia are exposed to the substance and the test is ended when 60% of the surviving control females produced 3 broods of young.

GLP compliance:

not specified

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material (migrated information):
No test surrogate or analogue material

Analytical monitoring:

no

Details on sampling:

Not applicable (no analytical monitoring)

Vehicle:

no

Details on test solutions:

Water accommodated fractions (WAFs) were prepraed on alternate days durinfg testing accoding to methods described by Anderson et al. (Anderson, 1974). Ten parts of the test water and one part of the test material were combined to prepare each WAF; actual volumes of liquids were dependent on the volume needed for testing. The solutions were mixed for 20 +/- 1 hours using a magnetic stir bar and a stir plate. The vortex produced during mixing was not allowed to exceed 25% of the distance from the liquid surface to the bottom of the mixing chamber. After mixing, the WAFs were extracted from a point near the bottom of the mixing chambers by siphoning.

Test organisms (species):

Ceriodaphnia dubia

Details on test organisms:

TEST ORGANISM
- Common name: Water flea
- Source: obtained from in-house cultures at the Fort Collins Environmental Toxicology Laboratory
- Age: At the time of test initiation, the test organisms were less than 24-hour old
- Feeding during test
- No feeding during the range-finding test
- Feeding during the definitive test:
- Food type: yeast-trout chow-cereal leaves/algae mix
- Amount: 0.2 ml per test chamber
- Frequency: daily


ACCLIMATION
- Acclimation period: 24 h after hatching
- Acclimation conditions: same as test
- Health during acclimation (any mortality observed): Good physical condition

Test type:

semi-static

Water media type:

freshwater

Limit test:

no

Total exposure duration:

6 d

Post exposure observation period:

Data were gathered through 7 days, although 60% of the surviving control females had third brood by day 6, when the test should have been terminated.

Hardness:

96-106 mg/L CaCO3

Test temperature:

20+/- 1°C

pH:

8.0-8.3

Dissolved oxygen:

>=6.9 mg/L

Salinity:

Conductivity: 339-346 µS/cm

Nominal and measured concentrations:

0, 12.5, 25, 50, 75 and 100% WAF

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 30 mL plastic beakers with 15 mL test medium
- No aeration during any of the tests
- Renewal rate of test solution (frequency/flow rate): daily
- No. of organisms per vessel: 1
- No. of vessels per concentration (replicates): 10
- No. of vessels per control (replicates):10


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Reconstituted moderatly hard water (USEPA, 1994. Short-term methods for estimating the chronic toxicity of effluents and receiving water to freshwater organisms. EPA/600/4-91/002)
- Chlorine <0.05 mg/L
- Alkalinity: 66-69 mg/L CaCO3
- Conductivity:339-364 µS/cm
- Intervals of water quality measurement: daily


OTHER TEST CONDITIONS
- Adjustment of pH: no
- Photoperiod:16 h L / 8 h D (fluorescent lighting)


EFFECT PARAMETERS MEASURED (with observation intervals if applicable) :
- Mortality of adults (defined as no visible movement after gentle prodding with a blunt probe
- Total number of young produced per adult


RANGE-FINDING STUDY
- Test concentrations: 0, 1, 10 and 100% WAF for 24 hours
- Results used to determine the conditions for the definitive study: the high concentration for the definitive test is set at 100% WAF.

Reference substance (positive control):

yes

Remarks:

Sodium chloride NaCl

Key result

Duration:

6 d

Dose descriptor:

EL50

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: Not reached

Duration:

6 d

Dose descriptor:

LOELR

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: No statistical differences between control and treated

Key result

Duration:

6 d

Dose descriptor:

NOELR

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

mortality

Remarks on result:

other: No statistical differences between control and treated

Key result

Duration:

6 d

Dose descriptor:

EL50

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: Not reached

Duration:

6 d

Dose descriptor:

LOELR

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: No statistical differences between control and treated

Key result

Duration:

6 d

Dose descriptor:

NOELR

Effect conc.:

> 100 other: % WAF

Nominal / measured:

nominal

Conc. based on:

test mat.

Basis for effect:

reproduction

Remarks on result:

other: No statistical differences between control and treated

Details on results:

- Mortality of parent animals: see table 1 (§ Remarks on results including tables and figures)
- No. of offspring produced per day per female: see table 1 (§ Remarks on results including tables and figures)

Results with reference substance (positive control):

IC25 = 554 mg/L (95% Confidence interval: 136-745) (NaCl as mg/L Cl-)

Reported statistics and error estimates:

See table 2 (§ Remarks on results including tables and figures)

Table 1. Chronic exposure ofCeriodaphnia dubiato Vista C13-C16: Definitve test results

 

Test concentration (% WAF)	Survival (%) on each test day						Mean young per female
	1	2	3	4	5	6
0 (Control)	100	100	100	100	90	90	18.0
12.5	100	100	100	100	100	100	16.8
25	100	100	100	90	90	90	15.7
50	100	100	100	100	90	90	13.0
75	100	100	100	100	100	90	15.8
100	100	100	100	100	100	100	10.6

 

 

Table 2. Chronic exposure ofCeriodaphnia dubiato Vista C13-C16: Statistical methods

 

End point	Comparison	Procedure
Survival	Significant reduction relative to the dilution water control	Inspection
	Treshold inhibition concentration (IC25/EL25)	Inspection
Reproduction	Normality	Chi-Square test (α=0.01)
	Homogeneity of variance	Bartlett’s test (α=0.01)
	Significant reduction relative to the dilution water control	Dunnett’s test (α=0.05)
	Treshold inhibition concentration (IC25/EL25)	Linear interpolation

Validity criteria fulfilled:

yes

Conclusions:

The NOELR for Ceriodaphnia dubia exposed to Vista C13-C16 was >100% WAF based on mortality and reproduction (no statistical differences between treatment groups)

Executive summary:

The study was performed to evaluate the toxicity of the water accommodated fractions (WAFs) of the test substance Vista C13-C16 to the water flea Ceriodaphnia dubia, in a semi-static system. The test was ended when 60% of the control females had produced 3 broods (6 days).

The water accommodated fractions were prepared by mixing 1 volume of the test substance with 10 volumes of dilution water. The dilution water was reconstituted moderately hard water prepared in the laboratory according to USEPA (USEPA, 1994. Short-term methods for estimating the chronic toxicity of effluents and receiving water to freshwater organisms. EPA/600/4-91/002). A range-finding test with 0, 1, 10 and 100 % WAF allowed setting the high concentration at 100% WAF for the definitive test. The definitive test was performed with 0, 12.5, 25, 50, 75 and 100 % WAF. The stock solution was prepared on alternate days as WAF by mixing 1 volume of test substance to 10 volumes of dilution water. The solution was mixed for 20±1 hours and the WAF was extracted from a point near the bottom of the mixing chamber by siphoning.

10 replicates, each containing one organism, were tested for each treatment. The test media were renewed daily. Mortality and reproduction were assessed daily.

 

An EL₂₅(reported as IC₂₅in the study report) was calculated for reproduction by linear interpolation:

EL₂₅= 80.9 % WAF

 

However, the statistical study showed that there was no statistical difference between treatment groups (including control group) for mortality as well as for reproduction. Therefore, the No Observed Effect Loading Rate based on adult mortality and reproduction (expressed as number of young per female) was set at > 100 % WAF.

NOELR = 100% WAF

Description of key information

There is no data available for this substance. However, key and weight-of-evidence data is available for structural analogues Hydrocarbons, C11-C13 (odd numbered), n-alkanes, <2% aromatics and Hydrocarbons, C11-C13, isoalkanes, <2% aromatics and Hydrocarbons, C12 -C15, n-alkanes, <2% aromatics. The data is read across to this substance based on analogue read across and a discussion and report on the read across strategy is provided as an attachment in IUCLID Section 13. Key and weight-of-evidence information is summarised below:

Hydrocarbons, C11-C13 (odd number), n-alkanes,<2% aromatics presented a 21-d NOELRs for Daphnia magna of ≥10.2mg/L (nominal loading rate), based on parent mortality and reproduction per surviving.

 

Hydrocarbons, C11-C13, isoalkanes, <2% aromatics presented a 21-d NOELRs for Daphnia magna of 1mg/L (nominal loading rate), based on effects on survival, reproduction, or length.

The NOELR for Ceriodaphnia dubia exposed to Hyd C12-C15, n-alkanes, <2% arom (Vista C13-C16) was >100% WAF based on mortality and reproduction (no statistical differences between treatment groups)

Additional information

Two studies are available investigating the long-term toxicity to aquatic invertebrates.

The study that tested Hydrocarbons, C11-C13 (odd numbered), n-alkanes, <2% aromatics to aquatic invertebrates was performed according to GLP and OECD guideline 211 using Daphnia magna as test organism (BASF, 2017). Loading rate WAFs between 0.52 and 10.2 mg/L were tested. There was no statistically significant effect on parent mortality and reproduction per surviving and introduced parent in any of the test groups. Thus, the 21 d-No Observed Effect Loading Rate (NOELR) is determined to be >= 10.2 mg/L based on nominal loading rate.

The study that tested Hydrocarbons, C11-C13, isoalkanes, <2% aromatics to aquatic invertebrates was performed according to GLP and OECD guideline 211 using Daphnia magna as test organism (ExxonMobil, 2001). The substance showed no effects on survival, reproduction, or length at a nominal loading of 1 mg/L. Therefore, the 21-Day NOELR is 1.0 mg/L for these endpoints based upon nominal loading levels.