Partially hydrogenated β-3,7,11-trimethyldodeca-1,3,6,10-tetraene, reaction products with linear C8-C16 alpha olefin, hydrogenated.

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

bioaccumulation: aquatic / sediment

Type of information:

calculation (if not (Q)SAR)

Adequacy of study:

supporting study

Study period:

Not specified

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

accepted calculation method

Justification for type of information:

Scientific evaluation and expert statement on the basis of:
Biodegradation test data
Log Pow Study data and calculations
Quantitative Structural Activity Relationships (QSAR)
Molecular Size Data
Metabolic Absorption test data
Literature data as appropriate.

Principles of method if other than guideline:

The substance is a hydrocarbon UVCB. The purpose of the report is to provide an assessment of the propensity of NovaSpec Renewable White Oil to bioaccumulate and prepare a suitable scientific comment paper for the purposes of supporting the notification of the material in various regulatory schemes (specifically China). The report utilises:

• Biodegradation test data
• Log Pow Study data and calculations
• Quantitative Structural Activity Relationships (QSAR)
• Molecular Size Data
• Metabolic Absorption test data
• Literature data as appropriate.

The outcome and results of the assessment are detailed below.

GLP compliance:

no

Specific details on test material used for the study:

Details on properties of test surrogate or analogue material:Not Applicable - Scientific evaluation and expert statement

Radiolabelling:

no

Details on sampling:

Not Applicable - Scientific evaluation and expert statement

Details on preparation of test solutions, spiked fish food or sediment:

Not Applicable - Scientific evaluation and expert statement

Details on test organisms:

Not Applicable - Scientific evaluation and expert statement

Route of exposure:

other: Scientific evaluation and expert statement

Test type:

other: Scientific evaluation and expert statement

Water / sediment media type:

not specified

Hardness:

Not Applicable - Scientific evaluation and expert statement

Test temperature:

Not Applicable - Scientific evaluation and expert statement

pH:

Not Applicable - Scientific evaluation and expert statement

Dissolved oxygen:

Not Applicable - Scientific evaluation and expert statement

TOC:

Not Applicable - Scientific evaluation and expert statement

Salinity:

Not Applicable - Scientific evaluation and expert statement

Conductivity:

Not Applicable - Scientific evaluation and expert statement

Details on test conditions:

Not Applicable - Scientific evaluation and expert statement

Nominal and measured concentrations:

Not Applicable - Scientific evaluation and expert statement

Reference substance (positive control):

not required

Details on estimation of bioconcentration:

The substance is a hydrocarbon UVCB. The purpose of the report is to provide an assessment of the propensity of NovaSpec Renewable White Oil to bioaccumulate and prepare a suitable scientific comment paper for the purposes of supporting the notification of the material in various regulatory schemes (specifically China). The report utilises:

• Biodegradation test data
• Log Pow Study data and calculations
• Quantitative Structural Activity Relationships (QSAR)
• Molecular Size Data
• Metabolic Absorption test data
• Literature data as appropriate.

The outcome and results of the assessment are detailed below.

Type:

BCF

Value:

3.162 - 1 944 L/kg

Basis:

whole body w.w.

Remarks on result:

other: Calculation by US EPA On-Line EPI Suite™ BCFBAF v3.01 model

Details on kinetic parameters:

Not Applicable - Scientific evaluation and expert statement

Metabolites:

Not Applicable - Scientific evaluation and expert statement

Results with reference substance (positive control):

Not Applicable - Scientific evaluation and expert statement

Details on results:

The following was evaluated:

Biodegradation test data
As the substance shows significant biodegradation under environmental conditions, it is not persistent. The absence of the substance within the environment for any considerable length of time therefore indicates that it is not available for uptake and subsequent bioaccumulation within aquatic organisms. As such, the substance is not considered to be potentially bioaccumulative on the consideration that it is not present within the environment for sufficient time to enable bioaccumulation to take place.

Log Pow Study data and calculations
The measured Log Kow is >5.31. Using the US EPA On-Line EPI Suite™ WSKOWWIN v1.42 model, the log Kow range is predicted to be 7.49 to 31.33. As the substance shows log Kow values in the main of > 10, it is proposed that the substance is not indicative of being potentially bioaccumulative, on the basis of the partition coefficient values observed.

Quantitative Structural Activity Relationships (QSAR)

Using the US EPA On-Line EPI Suite™ BCFBAF v3.01 model, the bioaccumulation range is predicted to be 3.162 to 1944 L/kg wet-wt. This is below the threshold for concern for bioaccumulation, and enforces the fact that significant uptake of the material in the mammalian studies was not observed. The substance is not considered to be potentially bioaccumulative.

Molecular Size Data

Evaluation of the selected molecules, taking into account the assumption that these are only a few of possible thousands of variants, indicates that approximately 29% of the molecules are of a Dmax average of greater than 17 Å (1.7 nm) plus a molecular weight of greater than 700 (Da). This indicates, within a weight of evidence approach and with expert judgment, that a good proportion of the substance is deemed to not be very bioaccumulative on the basis of physical size.

Metabolic Absorption test data

An assessment of metabolic absorption was investigated on the material using a single oral (gavage) administration at a dose level of 2000 mg/kg body weight. This dose did not cause any test item related effects. Analysis of the faeces showed the presence of the test item in the faeces at a range of approx. 34% - 63% of the administered weight. No test item was detected in the analyzed internal organs (gastrointestinal tract, liver, kidney and brain).

There was no evidence of any systemic absorption of the test item; the only test item detected was in the faeces. Recovery of the administered dose was 34%-63%. Recovery of this % amount is considered to be due to the analytical technique and is not indicative of absorption. The substance is not considered to be absorbed.


Literature data as appropriate.

Specific data on the bioaccumulation of white oils is limited with respect to substances like NovaSpec Renewable White Oil. This is presumably because materials of this nature are not expected to demonstrate a propensity towards bioaccumulation, based on their intrinsic properties.

Overall Conclusion.
The substance is a hydrocarbon UVCB. Assessment using the US EPA On-Line EPI Suite™ KOWWIN version 1.68 model, the log Kow range is predicted to be 7.49 to 31.33. As the substance shows log Kow values in the main of > 10, it is proposed that the substance is not indicative of being potentially bioaccumulative, on the basis of the partition coefficient values observed.
The substance is, however, considered to be not bioavailable to aqueous organisms as demonstrated by the lack of toxicity. Furthermore, the high log Kow is considered to be more a consequence of poor water solubility than a lipophilic tendency and is considered to be not indicative of the tendency to bioaccumulate in lipid tissues of aquatic organisms. This is based in part on an evaluation of literature data which demonstrates a tendency for the Bioconcentration Factor (BCF) to decrease as Log Pow increases above 6. This assumption is further confirmed by the data set available on the substance which demonstrates that it is poorly absorbed, both on the basis of the toxicity (or lack of) observed, plus the absorption study which demonstrated the lack significant absorption in the rat.
The above opinion is also confirmed via use of the EPIWIN BCFBAF v3.01 for assessment of bioaccumulation. The smallest and largest theoretical molecular weight substances were assessed and the following results were demonstrated:
Smallest: BCF 1944 L/kg wet-wt
Largest: BCF 3.162 L/kg wet-wt

All evaluated values are less that <2000, indicating that the substance is not proposed to be bioaccumulative.

Evaluation of predicted molecular size data indicates that approximately 29% of the molecules are of a Dmax average of greater than 17 Å (1.7 nm) plus a molecular weight of greater than 700 (Da).

Taking into account the following:

• Lack of absorption
• Lack of persistency
• High predicted log Kow value.
• Low predicted bioaccumulation potential
• Molecular size data not being indicative of the propensity towards bioaccumulation.


This indicates, within a weight of evidence approach and with expert judgment, that NovaSpec Renewable White Oil is deemed to not be bioaccumulative in aquatic organisms.

Reported statistics:

Not Applicable - Scientific evaluation and expert statement

The smallest and largest molecular weight substances were assessed to provide an indication of the propensity towards bioaccumulation. The results are below:

 

Component & compartment	Monomer MW = 212.41	Teramer (M-C16-C16-C16) MW = 885.69
Log BCF	3.289	0.500
BCF	1944 L/kg we-wt	3.162 L/kg wet-wt

Validity criteria fulfilled:

yes

Conclusions:

The substance is a hydrocarbon UVCB. Assessment using the US EPA On-Line EPI Suite™ KOWWIN version 1.68 model, the log Kow range is predicted to be 7.49 to 31.33. As the substance shows log Kow values in the main of > 10, it is proposed that the substance is not indicative of being potentially bioaccumulative, on the basis of the partition coefficient values observed.
The substance is, however, considered to be not bioavailable to aqueous organisms as demonstrated by the lack of toxicity. Furthermore, the high log Kow is considered to be more a consequence of poor water solubility than a lipophilic tendency and is considered to be not indicative of the tendency to bioaccumulate in lipid tissues of aquatic organisms. This is based in part on an evaluation of literature data which demonstrates a tendency for the Bioconcentration Factor (BCF) to decrease as Log Pow increases above 6. This assumption is further confirmed by the data set available on the substance which demonstrates that it is poorly absorbed, both on the basis of the toxicity (or lack of) observed, plus the absorption study which demonstrated the lack significant absorption in the rat.
The above opinion is also confirmed via use of the EPIWIN BCFBAF v3.01 for assessment of bioaccumulation. The smallest and largest theoretical molecular weight substances were assessed and the following results were demonstrated:
Smallest: BCF 1944 L/kg wet-wt
Largest: BCF 3.162 L/kg wet-wt

All evaluated values are less that <2000, indicating that the substance is not proposed to be bioaccumulative.

Evaluation of predicted molecular size data indicates that approximately 29% of the molecules are of a Dmax average of greater than 17 Å (1.7 nm) plus a molecular weight of greater than 700 (Da).

Taking into account the following:

• Lack of absorption
• Lack of persistency
• High predicted log Kow value.
• Low predicted bioaccumulation potential
• Molecular size data not being indicative of the propensity towards bioaccumulation.

This indicates, within a weight of evidence approach and with expert judgment, that NovaSpec Renewable White Oil is deemed to not be bioaccumulative in aquatic organisms.

Executive summary:

The substance is a hydrocarbon UVCB. Assessment using the US EPA On-Line EPI Suite™ KOWWIN version 1.68 model, the log Kow range is predicted to be 7.49 to 31.33.As the substance shows log Kow values in the main of > 10,it is proposed that the substance is not indicative of being potentially bioaccumulative, on the basis of the partition coefficient values observed.

The substance is, however, considered to be not bioavailable to aqueous organisms as demonstrated by the lack of toxicity. Furthermore, the high log Kow is considered to be more a consequence of poor water solubility than a lipophilic tendency and is considered to be not indicative of the tendency to bioaccumulate in lipid tissues of aquatic organisms. This is based in part on an evaluation of literature data which demonstrates a tendency for the Bioconcentration Factor (BCF) to decrease as Log Pow increases above 6. This assumption is further confirmed by the data set available on the substance which demonstrates that it is poorly absorbed, both on the basis of the toxicity (or lack of) observed, plus the absorption study which demonstrated the lack significant absorption in the rat.

The above opinion is also confirmed via use of the EPIWIN BCFBAF v3.01 for assessment of bioaccumulation. The smallest and largest theoretical molecular weight substances were assessed and the following results were demonstrated:

Smallest: BCF 1944 L/kg wet-wt

Largest: BCF 3.162 L/kg wet-wt

 

All evaluated values are less that <2000, indicating that the substance is not proposed to be bioaccumulative.

 

Evaluation of predicted molecular size data indicates that approximately 29% of the molecules are of a Dmax average of greater than 17 Å (1.7 nm) plus a molecular weight of greater than 700 (Da). 

 

Taking into account the following:

 

·        Lack of absorption

·        Lack of persistency

·        High predicted log Kow value.

·        Low predicted bioaccumulation potential

·        Molecular size data not being indicative of the propensity towards bioaccumulation.

  

This indicates, within a weight of evidence approach and with expert judgment, that NovaSpec Renewable White Oil is deemed to not be bioaccumulative in aquatic organisms.

Reference 2

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

key study

Study period:

May 18,2017 to June 15,2017

Reliability:

1 (reliable without restriction)

Rationale for reliability incl. deficiencies:

guideline study

Qualifier:

according to guideline

Guideline:

other: Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body

Version / remarks:

"Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body" stipulated in the "Testing Methods for New Chemical Substances" (March 31, 2011, No. 0331-7, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; March 29, 2011, No.5, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 110331009, Environmental Policy Bureau, Ministry of the Environment, Japan; latest revision, December 21, 2015, No. 1221-1, Pharmaceutical and Food Safety Bureau, Ministry ofHealth, Labour and Welfare; December 9, 2015, No.1, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 1512211, Environmental Policy Bureau, Ministry of the Environment, Japan)

Deviations:

not specified

GLP compliance:

yes

Specific details on test material used for the study:

No further details specified in the study report.

Radiolabelling:

no

Details on sampling:

Frequency of analysis
a) Test water analysis
The test water of Levels 1 and 2 were analyzed once before uptake phase and at the same time as analysis of test fish thereafter. In the blank tests, the test item was not detected in test water, therefore test water analysis of Control was performed only at the end of uptake phase. One sample was analyzed at each sampling time.
b) Test fish analysis
Analysis of test fish of Levels 1 and 2 was performed five times during the uptake phase. Four fish per treatment level were taken out at each sampling time and divided into two groups (two fish per group) because one fish was not enough for the analytical sensitivity of the test item. Test fish sampling was carried out at intervals of more than 48 hours, and the final analysis was performed after 28 days.
Analysis of Control fish was performed only after uptake phase. Four fish were taken out at the sampling time and divided into two groups (two fish per group). The test fish analysis before uptake phase was not conducted since the test item was not detected in the blank tests.

Vehicle:

yes

Details on preparation of test solutions, spiked fish food or sediment:

Acute toxicity test
Preparation of test solution
Test sample (500 mg) and HC)-40 (2.5 g) were dissolved in tetrahydrofuran to prepare 50.0 g/L stock solution (10 mL). This stock solution was diluted appropriately by dilution water to prepare the test solution.

Bioconcentration test
Preparation of stock solutions
a) Level 1
Test sample (1.00 g) and HCO-40 (5.0 g) were dissolved in tetrahydrofuran to prepare the 2000 mg/L stock solution of test item (500 mL).
b) Level 2
Test sample (100 mg) and HCO-40 (5.0 g) were dissolved in tetrahydrofuran to prepare the 200 mg/L stock solution of test item (500 mL).
c) Control
HCO-40 (20 g) was dissolved in tetrahydrofuran to prepare 10 g/L stock solution of HCO-40 (2 L).

Test organisms (species):

Cyprinus carpio

Details on test organisms:

Acute toxicity test
Species: Ricefish (Oryzias latipes)
Reason for selection: This species is similar in sensitivity to common carp and readily available as test fish.
Supplier: Kitamura fish farm (Address 388 Gunchiku 12-bancho, Y atsushiro-shi, Kumamoto 866-0001, Japan)
Lot No.: TFO-161206
Weight: 0.28-0.34 g
Length: 3.1-3.3 cm

Bioconcentration test
Species: Common carp (Cyprinus carpio)
Reason for selection: The previous data conducted with this species can be compared and the size of this species is adequate for handling.
Supplier: CERI Kurume
Conditions for acclimatization: The external disinfection was carried out in an aqueous solution containing OTC for fisheries (oxytetracycline hydrochloride, Kyoritsu Seiyaku) and sodium chloride (The Salt Industry Center of Japan).
Thereafter fish were acclimatized in the following conditions.
Period: 49 days
Temperature: 25 ± 2 °C
Mortality during acclimatization was less than 5%.
Lot No.: TFC-170221
Length: 6.7-8.3 cm (at the beginning of exposure 6.8-7.5 cm)
Age: Yearling fish

Feeding
Feed: Feed for colored carp
Composition: Proteins content 2:30.0%
Lipid content 2:4.0%
Manufacturer: KYORIN FOOD INDUSTRY LTD.
Feeding amount and interval: Amount corresponding to 2% of total body weight was fed twice a day in halves (once a day in all at holiday).
The fish were starved for 24 hours before sampling.

Route of exposure:

aqueous

Justification for method:

other: In accordance with test guidelines

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

28 d

Hardness:

Total hardness of test water
Level 1: 19.0 mg CaC03/L
Control: 17.0 mg CaC03/L

Test temperature:

Before uptake phase
Level 1: 24.3-24.4 °C; Level 2: 24.4-24.5 °C; Control: 24.7-24.8 °C
Uptake phase
Level 1: 24.0-24.6 °C; Level 2: 24.0-24.5 °C; Control: 24.2-24.8 °C

pH:

Before uptake phase
Level 1: 7.5; Level 2: 7.5; Control: 7.5
Uptake phase (at the beginning, and end of uptake phase)
Level 1: 7.4, 7.3; Level 2: 7.4, 7.2; Control: 7.4, 7.2

Dissolved oxygen:

Before uptake phase
Level 1: 7.3 mg/L; Level 2: 7.3 mg/L; Control: 7.3 mg/L
Uptake phase
Level 1: 6.7-7.1 mg/L; Level 2: 6.7-7.1 mg/L; Control: 6.8-7.2 mg/L

TOC:

Before uptake phase (before 48 hand 24 h)
Level 1: 22.4, 23.4 mgC/L; Level 2: 22. 7, 23.4 mgC/L; Control: 26.3, 21.0 mgC/L
Uptake phase
Level 1: 23.2-25.2mgC/L; Level 2: 22.4-23.7 mgC/L; Control: 18.2-22.3 mgC/L

Salinity:

Not applicable - freshwater study

Conductivity:

Not specified

Details on test conditions:

Conditions of test and circumstances
Supply of test water: Flow-through system assembled at this laboratory was used.
Test tank: 70-L glass tank
Level 1 No. 2-5
Level 2 No. 2-6
Control No. 2-7
Flow rate of test water: 0.04 mL/min for stock solution and 800 mL/min for dilution water, 1152 L/day of test water, were supplied from May 15, 2017.
Stock solution bottle: 0.5-L brown glass bottle (Levels 1 and 2) (Frequency of renewal: 1-2 times / week). 3-L brown glass bottle (Control) (Frequency of renewal: Once during the uptake phase)
Aeration: No aeration
Time of irradiation with light: 14 hours light /10 hours dark (artificial light of white fluorescent lamp)
Number of fish (at the beginning of exposure): Levels 1 and 2 26 fish; Control 16 fish
Duration of exposure: 28 days (May 18, 2017- June 15, 2017)
Reason: It was evaluated that a steady-state was reached after 28 days.

Observation, measurement and cleaning
Water quality was measured as following table. During the experiment, experimental excreta of carp, dirt on test tanks were removed about once a day.
Observation of test fish: Twice a day (once a day in holiday) during uptake phase
Water temperature: Daily before and during uptake phase
Flow rate of test water: Once a day before and during uptake phase
Dissolved oxygen: Once before the uptake phase, and once or twice a week during uptake phase
pH: Once before uptake phase, and twice during uptake phase
Total organic carbon: Twice before uptake phase (24h and 48h before the uptake phase) and, once a week during uptake phase
Total hardness: Once during the uptake phase for Control and Level 1

Nominal and measured concentrations:

Test nominal concentrations of the test item were set as follows. The control was set as a blank test.
Level 1: 0.1 mg/L
Level 2: 0.01 mg/L

Reference substance (positive control):

no

Details on estimation of bioconcentration:

Lipid content in test fish
Lipid contents of test fish was determined using control fish samples before and at the end of uptake phase. Six fish were taken out at each sampling time and divided into three groups (two fish per group). The each group was pretreated, and lipid contents were determined with gravimetric analysis after chloroform-methanol extraction.

Calculation of the test item concentration in sample and LOQ
LOQ of the test item in test water
The LOQ*4 of the test item in test water was calculated as follows:
Level 1
Peak 1 0.0065 mg/L
Peak 2 0.0037 mg/L
Peak 3 0.0050 mg/L
Level 2
Peak 2 0.00037 mg/L

LOQ of the test item in test fish
Assuming the fish to be 10 g, the LOQ*4 of the test item in test fish was calculated as follows:
Peak 1 5.6 μg/g
Peak 2 0.19 μg/g
Peak 3 2.6 μg/g

*4 LOQ of the test item (mg/L or μg/g) = [(B/100) x ({CxE}/D)] / A

A: The lowest quantification concentration of the test item (mg/L)
B: Recovery rate (%)
C: Sampling volume of test water (mL) or weight of test fish (g)
D: Final volume of sample solution (mL)
E: Ratio of the portion, used for analysis to whole volume
Results were rounded off to 2 figures.

Calculation of results
Calculation of bioconcentration factor (BCF)
BCF was calculated as follows.
Calculation of BCF

BCF=Cf/Cw

BCF: Bioconcentration factor (L/kg)
Cf: Concentration of the test item in test fish (subtract FB) (μg/g)
Cw: The average concentration of the test item in test water during uptake phase (mg/L)
FB: The average concentration of blank in analysis of control (μg/g)

The average BCF in m-th analysis

BCFm = (BCFa + BCFb) / n

BCFm: The average bioconcentration factor in m-th analysis (number of group 2 (a,b)) (L/kg)
BCFa,b: Bioconcentration factor in each group at m-th analysis (L/kg)
n: Number of group in m-th analysis of test fish
BCFm was not calculated when the test item concentration in test fish at m-th analysis was not higher than LOQ.

Definition of steady-state
The steady-state is defined to reach when the variation of concentration in test fish in four successive analyses at intervals of at least two days is within ± 20% of each other, and there is no significant increase of the test item concentration in test fish between the first and last successive analysis. When BCFs are less than 100 L/kg, it is evaluated that a steady-state has been reached after 28 days even if the variations of the test item concentration in test fish are over ± 20 %.
Criterion of the steady-state was reached: V(m-3),V(m-2), V(m-1), V(m) ≤ 20 (%)

V(m-3) = [(Cf(m-3) – Cf) / Cf] x 100
V(m-2) = [(Cf(m-2) – Cf) / Cf] x 100
V(m-1) = [(Cf(m-1) – Cf) / Cf] x 100
V(m) = [(Cf(m) – Cf) / Cf] x 100
V(m-3), V(m-2), V(m-1), V(m): Variation rate of test fish concentration from the average value (%)
Cf(m-3), Cf(m-2), Cf(m-1), Cf(m): The average concentration of the test item in test fish on m-3, m-2, m-1, m-th analysis of group n
Cf: {Cf(m-3) + Cf(m-2) + Cf(m-1) + Cf(m)} / 4

Calculable BCF
On the basis of the LOQ of the test item, calculable BCFs are as follows.
Level 1
Peak 1 63 L/kg
Peak 2 2.0 L/kg
Peak 3 29 L/kg
Level 2
Peak 2 19 L/kg

Calculation of growth rate constant (kg)
The individual measured fish weight data (n=6) in each level was used for the calculation of the growth rate constant (kg) in the study. The fish for the kg were used as follows.
The individual measured fish weight data were converted to natural logarithms and ln (fish weight) was plotted vs. time (day), then a linear least squares correlation was calculated for this plot. The kg was calculated as the slope of the linear correlation.
Beginning of uptake phase
Level 1, Level 2, Control: Six test fish for lipid content analysis before uptake phase
End of uptake phase
Level 1: Four test fish for test item analysis after 28 days + additional two test fish in Level 1.
Level 2: Four test fish for test item analysis after 28 days + additional two test fish in Level 2.
Control: Six test fish for lipid content analysis at the end of uptake phase in Control

Calculation of lipid content
Lipid contents were calculated according to following equation.
Lipid content (%) = (T- T0) / S x 100
T0: Weight of vessel (g)
T: Weight of sample for gravimetric analysis containing vessel (g)
S: Weight of homogenate sample taken out for analysis of lipid content (g)

Lipid content:

5.99 %

Time point:

start of exposure

Lipid content:

5.59 %

Time point:

end of exposure

Key result

Conc. / dose:

0.1 mg/L

Temp.:

>= 24 - <= 24.6 °C

pH:

7.3

Type:

BCF

Value:

< 63 L/kg

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Remarks on result:

other: Peak 1

Key result

Conc. / dose:

0.1 mg/L

Temp.:

>= 24 - <= 24.6 °C

pH:

7.3

Type:

BCF

Value:

< 2 L/kg

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Remarks on result:

other: Peak 2

Key result

Conc. / dose:

0.1 mg/L

Temp.:

>= 24 - <= 24.6 °C

pH:

7.3

Type:

BCF

Value:

< 29 L/kg

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Remarks on result:

other: Peak 3

Key result

Conc. / dose:

0.01 mg/L

Temp.:

>= 24 - <= 24.5 °C

pH:

7.2

Type:

BCF

Value:

< 19 L/kg

Basis:

whole body w.w.

Time of plateau:

28 d

Calculation basis:

steady state

Details on kinetic parameters:

Not specified

Metabolites:

Not specified

Results with reference substance (positive control):

Reference substance not required

Details on results:

Results of test fish observation: No abnormality in behavior or appearance was noted.

Reported statistics:

Not specified

Calculation table for recovery and blank test

(analysis of test water, peak 1)

Sample description	A	B	C	D	E	F
Standard 10.0 mg/L	34144
Recovery a	33987	1/1	1	-	9.95	99.5
Recovery b	32453	1/1	1	-	9.50	95.0
						Average 97.3
Standard 10.0 mg/L	32602
Blank a	n.d.	1/1	1	-	-
Blank b	n.d.	1/1	1	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis C: Final volume D: Amount of blank in test water (μg) E: Amount of test item recovered (μg)                E = P x (A(t) / A(std)) / B x C –D F: Recovery rate (%)                F = E / Q x 100 P: Concentration of test item in standard solution 10.0 mg/L Q: Amount of test item added (10.0 μg)

 

Calculation table for recovery and blank test

(analysis of test water, peak 2)

Sample description	A	B	C	D	E	F
Standard 10.0 mg/L	169316
Recovery a	147666	1/1	1	-	8.72	87.2
Recovery b	146711	1/1	1	-	8.66	86.6
						Average 86.9
Standard 10.0 mg/L	164274
Blank a	n.d.	1/1	1	-	-
Blank b	n.d.	1/1	1	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis C: Final volume D: Amount of blank in test water (μg) E: Amount of test item recovered (μg)                E = P x (A(t) / A(std)) / B x C –D F: Recovery rate (%)                F = E / Q x 100 P: Concentration of test item in standard solution 10.0 mg/L Q: Amount of test item added (10.0 μg)

 

Calculation table for recovery and blank test

(analysis of test water, peak 3)

Sample description	A	B	C	D	E	F
Standard 10.0 mg/L	24902
Recovery a	24649	1/1	1	-	9.90	99.0
Recovery b	25346	1/1	1	-	10.2	102
						Average 100
Standard 10.0 mg/L	45826
Blank a	n.d.	1/1	1	-	-
Blank b	n.d.	1/1	1	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis C: Final volume D: Amount of blank in test water (μg) E: Amount of test item recovered (μg)                E = P x (A(t) / A(std)) / B x C –D F: Recovery rate (%)                F = E / Q x 100 P: Concentration of test item in standard solution 10.0 mg/L Q: Amount of test item added (10.0 μg)

 

Calculation table for analysis of test water

(Level 1, peak 1)

Sample description	A	I
Standard 10.0 mg/L Before uptake phase	269381 218729	0.0835
Standard 10.0 mg/L Test water after 5 days	357426 299684	0.0862
Standard 10.0 mg/L Test water after 11 days	290522 255335	0.0903
Standard 10.0 mg/L Test water after 19 days	302870 262269	0.0890
Standard 10.0 mg/L Test water after 22 days	235222 197591	0.0863
Standard 10.0 mg/L Test water after 28 days	173183 154401	0.0916
Average concentration of test item in test water during uptake phase 0.0887 (S.D. 0.00241)
A: Peak area                A(std): Standard solution  A(t): Sample B: Ratio of portion used for analysis               1/1 C: Final volume  1 mL F: Recovery rate  97.3% H: Volume of test water taken out   100 mL I: Concentration of test item in test water (mg/L)                I = P x (A(t) / A(std) / B x C / F x 100 / H J: Average concentration of test item in test water during uptake phase (mg/L)                J = (I(1) + … + 1(n) / n                n: Number of test water analyses (n = 5)                I(1): First analysis of test water        I(n): Last analysis of test water                S.D. = √([n x ΣI(i)^2] x ΣI(i)]^2) / n x (n -1)                                i = 1              i = 1 P: Concentration of test item in standard solution       10.0 mg/L

 

Calculation table for analysis of test water

(Level 1, peak 2)

Sample description	A	I
Standard 10.0 mg/L Before uptake phase	1839170 1365215	0.0854
Standard 10.0 mg/L Test water after 5 days	2130070 1762411	0.0952
Standard 10.0 mg/L Test water after 11 days	1747709 1551433	0.102
Standard 10.0 mg/L Test water after 19 days	1752040 1463605	0.0961
Standard 10.0 mg/L Test water after 22 days	1426243 1140640	0.0920
Standard 10.0 mg/L Test water after 28 days	1461637 1208450	0.0951
Average concentration of test item in test water during uptake phase 0.0961 (S.D. 0.00370)
A: Peak area                A(std): Standard solution  A(t): Sample B: Ratio of portion used for analysis               1/1 C: Final volume  1 mL F: Recovery rate  86.9% H: Volume of test water taken out   100 mL I: Concentration of test item in test water (mg/L)                I = P x (A(t) / A(std) / B x C / F x 100 / H J: Average concentration of test item in test water during uptake phase (mg/L)                J = (I(1) + … + 1(n) / n                n: Number of test water analyses (n = 5)                I(1): First analysis of test water        I(n): Last analysis of test water                S.D. = √([n x ΣI(i)^2] x ΣI(i)]^2) / n x (n -1)                                i = 1              i = 1 P: Concentration of test item in standard solution       10.0 mg/L

 

Calculation table for analysis of test water

(Level 1, peak 3)

Sample description	A	I
Standard 10.0 mg/L Before uptake phase	242120 205986	0.0848
Standard 10.0 mg/L Test water after 5 days	293036 257896	0.0877
Standard 10.0 mg/L Test water after 11 days	307277 267175	0.0866
Standard 10.0 mg/L Test water after 19 days	317320 284374	0.0893
Standard 10.0 mg/L Test water after 22 days	232678 215684	0.0923
Standard 10.0 mg/L Test water after 28 days	225603 209981	0.0927
Average concentration of test item in test water during uptake phase 0.0897 (S.D. 0.00273)
A: Peak area                A(std): Standard solution  A(t): Sample B: Ratio of portion used for analysis               1/1 C: Final volume  1 mL F: Recovery rate  100% H: Volume of test water taken out   100 mL I: Concentration of test item in test water (mg/L)                I = P x (A(t) / A(std) / B x C / F x 100 / H J: Average concentration of test item in test water during uptake phase (mg/L)                J = (I(1) + … + 1(n) / n                n: Number of test water analyses (n = 5)                I(1): First analysis of test water        I(n): Last analysis of test water                S.D. = √([n x ΣI(i)^2] x ΣI(i)]^2) / n x (n -1)                                i = 1              i = 1 P: Concentration of test item in standard solution       10.0 mg/L

 

Calculation table for analysis of test water

(Level 2, peak 2)

Sample description	A	I
Standard 10.0 mg/L Before uptake phase	1839170 1569113	0.00981
Standard 10.0 mg/L Test water after 5 days	2130070 1784129	0.00964
Standard 10.0 mg/L Test water after 11 days	1747709 1617007	0.0106
Standard 10.0 mg/L Test water after 19 days	1752040 1531372	0.0101
Standard 10.0 mg/L Test water after 22 days	1426243 1135339	0.00916
Standard 10.0 mg/L Test water after 28 days	1461637 1172648	0.00923
Average concentration of test item in test water during uptake phase 0.00974 (S.D. 0.000618)
A: Peak area                A(std): Standard solution  A(t): Sample B: Ratio of portion used for analysis               1 C: Final volume  1 mL F: Recovery rate  86.9%% H: Volume of test water taken out   100 mL I: Concentration of test item in test water (mg/L)                I = P x (A(t) / A(std) / B x C / F x 100 / H J: Average concentration of test item in test water during uptake phase (mg/L)                J = (I(1) + … + 1(n) / n                n: Number of test water analyses (n = 5)                I(1): First analysis of test water        I(n): Last analysis of test water                S.D. = √([n x ΣI(i)^2] x ΣI(i)]^2) / n x (n -1)                                i = 1              i = 1 P: Concentration of test item in standard solution       10.0 mg/L

 

Calculation table for recovery and blank test

(analysis of test fish, for Peak 1 at Level 1)

Sample description	A	C	D	E	F	G
Standard 10.0 mg/L	54804
Recovery a	40959	2/100	2	-	747	74.7
Recovery b	39037	2/100	2	-	712	71.2
						Average 73.0
Standard 10.0 mg/L	65649
Blank a	n.d.	2/100	2	-	-
Blank b	n.d.	2/100	2	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               1/1 C: Ratio of portion used for analysis (extracted solution) D: Final volume (mL) E: Amount of blank in test fish (μg) F: Amount of test item recovered (μg)                F = P x (A(t) / A(std)) / B / C x D – E G: Recovery rate (%)                G = F / Q x 100 P: Concentration of test item in standard solution      10.0 mg/L Q: Amount of test item added (1000 μg)

 

Calculation table for recovery and blank test

(analysis of test fish, for Peak 2 at Levels 1 and 2)

Sample description	A	C	D	E	F	G
Standard 10.0 mg/L	243825
Recovery a	182015	20/100	2	-	74.6	74.6
Recovery b	177866	20/100	2	-	72.9	72.9
						Average 73.8
Standard 10.0 mg/L	230310
Blank a	n.d.	20/100	2	-	-
Blank b	n.d.	20/100	2	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               1/1 C: Ratio of portion used for analysis (extracted solution) D: Final volume (mL) E: Amount of blank in test fish (μg) F: Amount of test item recovered (μg)                F = P x (A(t) / A(std)) / B / C x D – E G: Recovery rate (%)                G = F / Q x 100 P: Concentration of test item in standard solution      10.0 mg/L Q: Amount of test item added (100 μg)

 

Calculation table for recovery and blank test

(analysis of test fish, for Peak 3 at Level 1)

Sample description	A	C	D	E	F	G
Standard 10.0 mg/L	38684
Recovery a	30089	2/100	2	-	778	77.8
Recovery b	30072	2/100	2	-	777	77.7
						Average 77.8
Standard 10.0 mg/L	39498
Blank a	n.d.	2/100	2	-	-
Blank b	n.d.	2/100	2	-	-	-
				Average -
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               1/1 C: Ratio of portion used for analysis (extracted solution) D: Final volume (mL) E: Amount of blank in test fish (μg) F: Amount of test item recovered (μg)                F = P x (A(t) / A(std)) / B / C x D – E G: Recovery rate (%)                G = F / Q x 100 P: Concentration of test item in standard solution      10.0 mg/L Q: Amount of test item added (1000 μg)

 

Calculation table for analysis of test fish

(Level 1, peak 1)

Sample description	A	D	G	K	J	M
Standard 10.0 mg/L Test fish after 5 days a Test fish after 5 days b	205523 n.d. n.d.	1 1	9.40 8.45	- -	- -	-
Standard 10.0 mg/L Test fish after 11 days a Test fish after 11 days b	160865 n.d. n.d.	1 1	9.27 7.86	- -	- -	-
Standard 10.0 mg/L Test fish after 19 days a Test fish after 19 day b	183971 n.d. n.d.	1 1	8.78 9.98	- -	- -	-
Standard 10.0 mg/L Test fish after 22 days a Test fish after 22 days b	145036 n.d. n.d.	1 1	9.39 9.40	- -	- -	-
Standard 10.0 mg/L Test fish after 28 days a Test fish after 28 days b	144410 n.d. n.d.	1 1	9.80 11.0	- -	- -	-
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               2/100 C: Final volume                  2 mL D: Dilution factor E: Average concentration of blank in analysis of blank test      0 μg/g F: Recovery rate  73.0% G: Weight of test fish (g) K: Concentration of test item in test fish (μg/g)                K = {P x (A(t) / A(std)) / B x D x C / G – E} / F x 100 H: Average concentration of test item in test water (mg/L)        0.0887 J: BCF (L/kg)                J = K / H M: Average values of BCF(a) and BCF(b) (L/kg)                M = {BCF(a) + BCF(b)} / 2 P: Concentration of test item in standard solution      10.0 mg/L

 

Calculation table for analysis of test fish

(Level 1, peak 2)

Sample description	A	D	G	K	J	M
Standard 10.0 mg/L Test fish after 5 days a Test fish after 5 days b	4044306 n.d. n.d.	1 1	9.40 8.45	- -	- -	-
Standard 10.0 mg/L Test fish after 11 days a Test fish after 11 days b	518019 n.d. n.d.	1 1	9.27 7.86	- -	- -	-
Standard 10.0 mg/L Test fish after 19 days a Test fish after 19 day b	426493 n.d. n.d.	1 1	8.78 9.98	- -	- -	-
Standard 10.0 mg/L Test fish after 22 days a Test fish after 22 days b	419628 n.d. n.d.	1 1	9.39 9.40	- -	- -	-
Standard 10.0 mg/L Test fish after 28 days a Test fish after 28 days b	1170532 n.d. n.d.	1 1	9.80 11.0	- -	- -	-
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               20/100 C: Final volume                  2 mL D: Dilution factor E: Average concentration of blank in analysis of blank test      0 μg/g F: Recovery rate  73.8% G: Weight of test fish (g) K: Concentration of test item in test fish (μg/g)                K = {P x (A(t) / A(std)) / B x D x C / G – E} / F x 100 H: Average concentration of test item in test water (mg/L)        0.0961 J: BCF (L/kg)                J = K / H M: Average values of BCF(a) and BCF(b) (L/kg)                M = {BCF(a) + BCF(b)} / 2 P: Concentration of test item in standard solution      10.0 mg/L

 

Calculation table for analysis of test fish

(Level 1, peak 3)

Sample description	A	D	G	K	J	M
Standard 10.0 mg/L Test fish after 5 days a Test fish after 5 days b	297857 n.d. n.d.	1 1	9.40 8.45	- -	- -	-
Standard 10.0 mg/L Test fish after 11 days a Test fish after 11 days b	235115 n.d. n.d.	1 1	9.27 7.86	- -	- -	-
Standard 10.0 mg/L Test fish after 19 days a Test fish after 19 day b	257205 n.d. n.d.	1 1	8.78 9.98	- -	- -	-
Standard 10.0 mg/L Test fish after 22 days a Test fish after 22 days b	208152 n.d. n.d.	1 1	9.39 9.40	- -	- -	-
Standard 10.0 mg/L Test fish after 28 days a Test fish after 28 days b	158834 n.d. n.d.	1 1	9.80 11.0	- -	- -	-
(a,b: individual sample)
A: Peak area            A(std): Standard solution          A(t): sample B: Ratio of portion used for analysis     2/100 C: Final volume            2 mL D: Dilution factor E: Average concentration of blank in analysis of blank test        0 μg/g F: Recovery rate           77.8% G: Weight of test fish (g) K: Concentration of test item in test fish (μg/g)            K = {P x (A(t) / A(std)) / B x D x C / G – E} / F x 100 H: Average concentration of test item in test water (mg/L)          0.0897 J: BCF (L/kg)            J = K / H M: Average values of BCF(a) and BCF(b) (L/kg)            M = {BCF(a) + BCF(b)} / 2 P: Concentration of test item in standard solution         10.0 mg/L

 

Calculation table for analysis of test fish

(Level 2, peak 2)

Sample description	A	D	G	K	J	M
Standard 10.0 mg/L Test fish after 5 days a Test fish after 5 days b	332890 n.d. n.d.	1 1	8.35 8.40	- -	- -	-
Standard 10.0 mg/L Test fish after 11 days a Test fish after 11 days b	518686 n.d. n.d.	1 1	8.00 8.67	- -	- -	-
Standard 10.0 mg/L Test fish after 19 days a Test fish after 19 day b	618115 n.d. n.d.	1 1	8.82 8.79	- -	- -	-
Standard 10.0 mg/L Test fish after 22 days a Test fish after 22 days b	499534 n.d. n.d.	1 1	9.76 8.69	- -	- -	-
Standard 10.0 mg/L Test fish after 28 days a Test fish after 28 days b	823764 n.d. n.d.	1 1	10.0 11.0	- -	- -	-
(a,b: individual sample)
A: Peak area                A(std): Standard solution  A(t): sample B: Ratio of portion used for analysis               20/100 C: Final volume                  2 mL D: Dilution factor E: Average concentration of blank in analysis of blank test      0 μg/g F: Recovery rate  73.8% G: Weight of test fish (g) K: Concentration of test item in test fish (μg/g)                K = {P x (A(t) / A(std)) / B x D x C / G – E} / F x 100 H: Average concentration of test item in test water (mg/L)        0.00974 J: BCF (L/kg)                J = K / H M: Average values of BCF(a) and BCF(b) (L/kg)                M = {BCF(a) + BCF(b)} / 2 P: Concentration of test item in standard solution      10.0 mg/L

 

Validity criteria fulfilled:

yes

Conclusions:

Acute toxicity test
96-hour LC50 >10.0 mg/L

Bioconcentration test
BCF
Level 1 (0.1 mg/L)
Peak 1: <63 L/kg
Peak 2: <2.0 L/kg
Peak 3: <29 L/kg

Level 2 (0.01 mg/L)
Peak 2: <10 L/kg

Executive summary:

Test item: NovaSpec base oil

Objective: This study is performed to evaluate the bioconcentration potential of NovaSpec base oil in common carp.

Test method: "Method for Testing the Degree of Accumulation of Chemical Substances in Fish Body" stipulated in the "Testing Methods for New Chemical Substances" (March 31, 2011, No. 0331-7, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare; March 29, 2011, No. 5, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 110331009, Environmental Policy Bureau, Ministry of the Environment, Japan; latest revision December 21, 2015, No. 1221-1, Pharmaceutical and Food Safety Bureau, Ministry of Health, Labour and Welfare;

December 9, 2015, No. 1, Manufacturing Industries Bureau, Ministry of Economy, Trade and Industry; No. 1512211, Environmental Policy Bureau, Ministry of the Environment, Japan)

 

Test conditions

a) Acute toxicity test

Test fish: Ricefish (Oryzias latipes)

Duration of exposure: 96 hours

Exposure method: Semi static system (Renewal of test water, at every 8 - 16 hours)

 

b) Bioconcentration test

Test fish: Common carp (Cyprinus carpio)

Nominal concentrations of test item:

High exposure level (Level 1): 0.1 mg/L

 

Low exposure level (Level 2): 0.01 mg/L

Duration of exposure: 28 days

Exposure method: Flow-through system

Analytical method: Gas chromatography-mass spectrometry

 

Results

a) Acute toxicity test

96-hour LC50 value >10.0 mg/L

 

b) Bioconcentration test

	Peak	Bioconcentration factor (L/kg)
Level 1	1	<63
	2	<2.0
	3	<29
Level 2	2	<10

 

Description of key information

Bioaccumulation study.

Key value for chemical safety assessment

BCF (aquatic species):

63 L/kg ww

Additional information

A full bioaccumulation study in accordance with Japanese Guidelines provided the following results:

Bioconcentration test

BCF

Level 1 (0.1 mg/L)

Peak 1: <63 L/kg

Peak 2: <2.0 L/kg

Peak 3: <29 L/kg

Level 2 (0.01 mg/L)

Peak 2: <10 L/kg

It was evaluated that a steady-state was reached within 28 days because all BCFs were less

than 100 L/kg.

The substance is a hydrocarbon UVCB. Assessment using the US EPA On-Line EPI Suite™ KOWWIN version 1.68 model, the log Kow range is predicted to be 7.49 to 31.33.As the substance shows log Kow values in the main of > 10,it is proposed that the substance is not indicative of being potentially bioaccumulative, on the basis of the partition coefficient values observed.

The substance is, however, considered to be not bioavailable to aqueous organisms as demonstrated by the lack of toxicity. Furthermore, the high log Kow is considered to be more a consequence of poor water solubility than a lipophilic tendency and is considered to be not indicative of the tendency to bioaccumulate in lipid tissues of aquatic organisms. This is based in part on an evaluation of literature data which demonstrates a tendency for the Bioconcentration Factor (BCF) to decrease as Log Pow increases above 6. This assumption is further confirmed by the data set available on the substance which demonstrates that it is poorly absorbed, both on the basis of the toxicity (or lack of) observed, plus the absorption study which demonstrated the lack significant absorption in the rat.

The above opinion is also confirmed via use of the EPIWIN BCFBAF v3.01 for assessment of bioaccumulation. The smallest and largest theoretical molecular weight substances were assessed and all evaluated values are less that <2000, indicating that the substance is not proposed to be bioaccumulative.

 Evaluation of predicted molecular size data indicates that approximately 29% of the molecules are of a Dmax average of greater than 17 Å (1.7 nm) plus a molecular weight of greater than 700 (Da). 

 Taking into account the following:

The actual test data.

·       Demonstrated lack of absorption

·       Demonstrated lack of persistency

·       High predicted log Kow value.

·       Demonstrated lack of bioaccumulation potential

·       Molecular size data not being indicative of the propensity towards bioaccumulation.

  

This indicates, within a weight of evidence approach and with expert judgment, that NovaSpec Renewable White Oil is deemed to not be bioaccumulative in aquatic organisms.