Soaps, stocks, vegetable-oil, acidulated

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Reference 1

Endpoint:

bioaccumulation in aquatic species, other

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Justification for type of information:

The study record represents the experimental result on a substance which represents one chain length of the glyceride and fatty acid constituents of the 'Soaps, stocks, vegetable oil, acidulated’.

Principles of method if other than guideline:

Fish, algae and activated sludge were exposed to the test substance for 3 d, 1 d and 5 d respectively and bioaccumulation factors were estimated as a result.

GLP compliance:

no

Test organisms (species):

other: Leuciscus idus melanotus, Chlorella fusca var.vacuolata and activated sludge

Route of exposure:

aqueous

Test type:

static

Water / sediment media type:

natural water: freshwater

Nominal and measured concentrations:

Nominal concentration: 0.05 mg/L

Details on estimation of bioconcentration:

- Bioaccumulation of tristearin and palmitic acid was measured in fish following exposure to the substances for 3 d. The bioconcentration was measured as the concentration of chemical in the fish/average concentration of chemical in water (µg/g).
- Bioaccumulation of tristearin and palmitic acid was measured in algae and activated sludge following exposure to the substances for 1 d and 5 d respectively. The bioconcentration was measured as the concentration of chemical in algae or sludge/final concentration of chemical in water (µg/g).

Key result

Conc. / dose:

0.05 mg/L

Type:

BCF

Value:

10 dimensionless

Basis:

whole body w.w.

Remarks on result:

other: Value for tristearin in fish

Key result

Conc. / dose:

0.05 mg/L

Type:

BCF

Value:

60 dimensionless

Remarks on result:

other: Value for palmitic acid in fish

Details on results:

- The BCF values for tristearin in fish, algae and activated sludge were determined to be <10, 5,840 and 3,600 respectively.
- The BCF values for palmitic acid in fish, algae and activated sludge were determined to be 60, 8,400 and 2,800 respectively.
The differences observed between fish and algae/activated sludge values respectively suggests a detoxification process in higher developed organisms as fish (metabolism).

Comparing and extrapolating these short term results leads to the following bioaccumulation classification based on long-term tests (carried in accordance with the OECD flow-through-test):

- No or low accumulation: 1,000

- Medium level accumulation: 1,000 -10,000

- High accumulation: 10,000

Based on this, tristearin and palmitic acid both can be considered to have low bioaccumulation potential in fish.

Conclusions:

Under the conditions of the study, both tristearin and palmitic acid can be considered to have a low bioaccumulation potential in fish.

Executive summary:

The bioaccumulation of tristearin and palmitic acid was measured in fish, algae and activated sludge following exposure for 3, 1 and 5 d respectively. The bioconcentration in fish was measured as the concentration of chemical in the fish/average concentration of chemical in water (µg/g) while the bioconcentration in algae and activated sludge was measured as the concentration of chemical in algae or sludge/final concentration of chemical in water (µg/g). The BCF values for tristearin in fish, algae and activated sludge were determined to be <10, 5,840 and 3,600 respectively. The corresponding values for palmitic acid in fish, algae and activated sludge were determined to be 60, 8,400 and 2,800 respectively. The differences observed between fish and algae/activated sludge values respectively suggests a detoxification process in higher developed organisms as fish (metabolism). Comparing and extrapolating the short term results to long term values for both the substances leads to 'no or low' bioaccumulation classification in fish and 'medium' bioaccumulation classification in algae and activated sludge. Under the conditions of the study, both tristearin and palmitic acid can be considered to have a low bioaccumulation potential in fish (Freitag, 1985).

Reference 2

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

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

Reason / purpose for cross-reference:

reference to other study

Qualifier:

no guideline followed

Principles of method if other than guideline:

The activity of carboxylesterase (CaE), a class of non-specific serine hydrolases, was evaluated in vitro in tissues and microsomes of rainbow trout. In the assays the formation of 4-nitrophenol from 4-nitrophenyl acetate was measured spectrophotometrically.

GLP compliance:

no

Test organisms (species):

Oncorhynchus mykiss (previous name: Salmo gairdneri)

Details on test organisms:

TEST ORGANISM
- Common name: rainbow trout
- Source: trouts were obtained as eyed embryos from Mt. Lassen Trout Farms, Mt. Lassen CA, USA
- Age at study initiation: < 1 year
- Length at study initiation (length definition, mean, range and SD):
- Weight at study initiation: 1.64 ± 0.07 g wet weight
- Method of holding: Trout were held in flow-through aerated raceways at 12 ± 1 °C. The laboratory water was softened Lake Huron water that had been sand-filtered, pH adjusted with CO2, carbon-filtered, and ultraviolet irradiated. Laboratory water was monitored weekly for pH, alkalinity, conductivity, and hardness; and quarterly for selected inorganics, pesticides, and poly-chlorinated biphenyls. Typical water quality values were pH of 7.5, alkalinity of 43 mg/L, hardness of 70 mg/L (as CaCO3 ), and conductivity of 140 mhos/cm. Fish were killed by a blow to the head and placed immediately on ice before tissue preparation.

Route of exposure:

other: In vitro exposure

Test type:

other: In vitro study

Water / sediment media type:

natural water: freshwater

Key result

Remarks on result:

other: Based on the study results, glyceride ester constituents of the substance may be considered to be broken down by the CaE enzymes present in fish leading to their removal and low bioaccumulation.

- The results demonstrate that:

- Rainbow trout had high esterase activity over a broad range of temperatures. This may be favourable adaptation for fish such as rainbow trout which experience wide range of environmental temperature from 2 to 20 deg C.

- CaE activity significantly increased between the yolk-sac and juvenile life stages, but was not significantly different in juvenile and adult life stages of rainbow trout.

- Variation between the CaE activity in trout and three other families of freshwater fish was limited indicating similar activity in many fish species.

Further, a previous study with limited number of ester substrates indicated that fish have high CaE activity in both sera and liver (Lech and Melancon, 1980) and ester hydrolysis significantly reduced accumulation of several hydrophobic esters (Rodgers and Stalling, 1972; Barron et al., 1989; Barron et al., 1990).

Conclusions:

Based on the study results, the glyceride ester constituents of ‘Soaps, stocks, vegetable oil, acidulated’ may be considered to be broken down by the CaE enzymes present in fish leading to their removal and low bioaccumulation.

Executive summary:

The activity of carboxylesterase (CaE), a class of nonspecific serine hydrolases, was evaluated in vitro in tissues and microsomes of rainbow trout. In the assays the formation of 4-nitrophenol from 4-nitrophenyl acetate was measured spectrophotometrically. The results of this study demonstrated that rainbow trout had high esterase activity over a broad range of temperatures, that carboxylesterase (CaE) activity significantly increased between the yolk-sac and juvenile life stages, and that variation between the CaE activity in trout and three other families of freshwater fish was limited. Further, a previous study with limited number of ester substrates indicated that fish have high CaE activity in both sera and liver (Lechand Melancon, 1980) and ester hydrolysis significantly reduced accumulation of several hydrophobic esters (Rodgers and Stalling, 1972; Barron et al., 1989; Barron et al., 1990). Based on the above study results, the glyceride ester constituents of the ‘ Soaps, stocks, vegetable oil, acidulated’ may be considered to be broken down by the CaE enzymes present in fish leading to their removal and low bioaccumulation (Barron, 1999).

Reference 3

Endpoint:

bioaccumulation in aquatic species, other

Type of information:

other: review article

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

data from handbook or collection of data

Principles of method if other than guideline:

Review article describing the metabolism and functions of lipids and fatty acids in fish. Both type of substances play major roles as source of metabolic energy in processes as growth and reproduction.

Test organisms (species):

other: fish

Key result

Remarks on result:

other: Triglycerides or lipids and their constituent fatty acids are not considered to be bioaccumulative in fish

Fatty acid catabolism is the predominant source of energy in many species of fish. The catabolism of fatty acids occurs in the cellular organelles, mitochondria (and peroxisomes). The process is termed beta-oxidation and involves the sequential cleavage of two-carbon units, released as acetyl-CoA, through a cyclic series of reactions catalyzed by several distinct enzyme activities. Activated fatty acids are transported to the mitochondrion in the form of fatty acyltransferase, converted back into fatty acyl-CoA derivatives, and then undergo a round of dehydrogenation, hydration, second hydrogenation, and cleavage steps to produce acetyl-CoA and NADH. Then, the acetyl-CoA can be metabolized via the tricarboxylic cycle to produce more NADH. The NADH will eventually lead to the release of ATP through the process of oxidative phosphorilation, available to be used as energy source.

Fatty acid oxidation is an important source of energy in several tissues in fish (heart, red muscles, etc). Furthermore, fatty acids are the major source of metabolic energy in the development from egg to adult fish and also during reproduction and lipid depletion processes, such as migrations. They are also involved in the maintainance of the structure and function of cellular biomembranes (as part of phosphoglycerides).

Conclusions:

Considering the triglycerides and their constituent fatty acids to be metabolised via beta-oxidation to serve as an important source of metabolic energy for growth and reproduction, these consitutents are not considered to bioaccumulate in fish.

Executive summary:

Fatty acid catabolism is the predominant source of energy in many species of fish. The catabolism of fatty acids occurs in the cellular organelles, mitochondria (and peroxisomes). The process is termed beta-oxidation and involves the sequential cleavage of two-carbon units, released as acetyl-CoA, through a cyclic series of reactions catalyzed by several distinct enzyme activities. Activated fatty acids are transported to the mitochondrion in the form of fatty acyltransferase, converted back into fatty acyl-CoA derivatives, and then undergo a round of dehydrogenation, hydration, second hydrogenation, and cleavage steps to produce acetyl-CoA and NADH. Then, the acetyl-CoA can be metabolized via the tricarboxylic cycle to produce more NADH. The NADH will eventually lead to the release of ATP through the process of oxidative phosphorylation, available to be used as energy source. Fatty acid oxidation is an important source of energy in several tissues in fish (heart, red muscles, etc). Furthermore, fatty acids are the major source of metabolic energy in the development from egg to adult fish and also during reproduction and lipid depletion processes, such as migrations. They are also involved in the maintenance of the structure and function of cellular biomembranes (as part of phosphoglycerides). Therefore, considering the triglycerides and their constituent fatty acids to be metabolised via beta-oxidation to serve as an important source of metabolic energy for growth and reproduction, these constituents are not considered to bioaccumulate in fish (Tocher, 2003).

Reference 4

Endpoint:

bioaccumulation: aquatic / sediment

Type of information:

(Q)SAR

Adequacy of study:

key study

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

results derived from a valid (Q)SAR model and falling into its applicability domain, with adequate and reliable documentation / justification

Justification for type of information:

(Q)SAR estimation according to BCFBAF v3.01 of EPISuite, CAESAR (BCF model) v.2.1.14 and T.E.S.T US EPA model v.4.1

Reason / purpose for cross-reference:

reference to other study

Qualifier:

according to guideline

Guideline:

other: REACH Guidance on QSARs R.6, May/July 2008

Principles of method if other than guideline:

1. Meylan,WM, Howard,PH, Boethling, RS et al. (1999). Improved Method for Estimating Bioconcentration / Bioaccumulation Factor from Octanol/Water Partition Coefficient. Environ. Toxicol. Chem. 18(4): 664-672.
2. Zhao C, Boriani E, Chana A, Roncaglioni A and Benfenati E (2008). A new hybrid system of QSAR models for predicting bioconcentration factors
(BCF)", Chemosphere 73:1701–1707.
3. Hamelink, J. L. 1977. Current bioconcentration test methods and theory. In Aquatic Toxicology and Hazard Evaluation, edited by F. L. Mayer and J. L. Hamelink. West Conshohocken, PA ASTM STP.

GLP compliance:

no

Vehicle:

no

Test organisms (species):

other: Not applicable

Details on test organisms:

Not applicable

Route of exposure:

other: Not applicable

Test type:

other: Not applicable

Reference substance (positive control):

not required

Details on estimation of bioconcentration:

In absence of experimental data, the bioaccumulation potential (BCF) of the test substance was determined through computational methods (such as BCFBAF v3.01 of EPIWEB v 4.1, BCF (CAESAR) 2.1.14 and T.E.S.T US EPA model v.4.1).

Assumption:
- The individual BCF values was calculated for the shortest and longest alkyl chain containing representative substances of each class present in the test substance.
- These representative substances each should represent at >10 % of the test substance composition.
- As the test substance is a UVCB the individual or average estimated Log Kow values for the constituents will be used to represent a range of values for the entire substance.

Therefore, the BCF values were determined for the following individual substances, present at >10% :
a) Octanoic acid, stearic and oleic acid representing free fatty acids (substance class)
The calculation input will be provided in SMILES notation as
Octanoic acid: O=C(O)CCCCCCC
Stearic acid: O=C(O)CCCCCCCCCCCCCCCCC
Oleic acid: O=C(O)CCCCCCCC=CCCCCCCCC
b) Triglyceride of Octanoic acid, stearic and oleic acid, representing glycerides of fatty acids (substance class)
The calculation input will be provided in SMILES notation as
Trioctanoin: CCCCCCCC(=O)OCC(COC(=O)CCCCCCC)OC(=O)CCCCCCC
Tristearin: O=C(CCCCCCCCCCCCCCCCC)OC(COC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC
Triolein: O=C(OC(COC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC)CCCCCCCC=CCCCCCCCC

- Method details:
1. BCFBAF v3.01 program of EPISuite v 4.1:
- For the triglycerides, the BCF values was estimated by using the derived QSAR estimation equation for Log Kow > 7.0 from EPISuite:
Log BCF = -0.49 Log Kow + 7.554 + Σ correction factors
(n = 35, r2 = 0.634, Q2 = 0.57, std dev = 0.538, avg dev = 0.396)
- For the fatty acids, the below equation for ionic compounds was used for the BCF estimation
Log BCF = 1.00 (Ionic; Log Kow dependent)


2. CAESAR v.2.1.14:
The BCF value via CAESAR is estimated using a combination of 2 Radial Basis Function Neural Network (RBF-NN) models (model A and B) developed with 5 descriptors each, for a total of 8 descriptors (2 are in common between the models). Details about the NN architecture are provided in the supporting information of the paper by Zhao et al., 2008. The estimations are categorised into the following scenarios:

- If mean (value given by models A and B) > 2.410: log BCF = 1.052 * [mean (value given by models A and B)] - 0.065
- If 1.355 < mean (value given by models A and B) ≤ 2.410: log BCF = 0.996 * [min (value given by models A and B)] + 0.042
- Otherwise log BCF = 0.936 * [mean (value given by models A and B)] - 0.123

3. T.E.S.T US EPA model v.4.1:
This model uses multiple methodologies for prediction of endpoints i.e.,
- Hierarchical clustering: The toxicity for a given query compound is estimated using the weighted average of the predictions from several different models. The different models are obtained by using Ward’s method to divide the training set into a series of structurally similar clusters. A genetic algorithm-based technique is used to generate models for each cluster. The models are generated prior to runtime.
- Single model method: Predictions are made using a multilinear regression model that is fit to the training set (using molecular descriptors as independent variables) using a genetic algorithm-based approach. The regression model is generated prior to runtime.
- Group contribution: Predictions are made using a multilinear regression model that is fit to the training set (using molecular fragment counts as independent variables). The regression model is generated prior to runtime.
- FDA method: The prediction for each test chemical is made using a new model that is fit to the chemicals that are most similar to the test compound. Each model is generated at runtime.
- Nearest neighbour: The predicted toxicity is estimated by taking an average of the three chemicals in the training set that are most similar to the test chemical.
Further details of these methodologies can be found in the publications mentioned in the US EPA website: https://www.epa.gov/chemical-research/toxicity-estimation-software-tool-test

Key result

Type:

other: Estimated BCF of fatty acids

Value:

ca. 11 - ca. 17 L/kg

Basis:

whole body w.w.

Calculation basis:

other: see details on results

Remarks on result:

other: Average BCF values estimated using BCFBAF of EPISuite, T.E.S.T. US EPA and CAESAR BCF models

Key result

Type:

other: Estimated BCF of glycerides

Value:

ca. 4 - ca. 143 L/kg

Basis:

whole body w.w.

Calculation basis:

other: see details on results

Remarks on result:

other: Average BCF values estimated using BCFBAF of EPISuite, T.E.S.T. US EPA and CAESAR BCF models

Key result

Type:

other: Overall estimated BCF for test substance

Value:

ca. 4 - ca. 143 L/kg

Basis:

whole body w.w.

Calculation basis:

other: see details on results

Remarks on result:

other: Based on average BCF values estimated for the individual constituents using BCFBAF of EPISuite, T.E.S.T. US EPA and CAESAR BCF models

Details on results:

Estimation by BCFBAF v3.01 program of EPIWEB v 4.1:
(A) Glycerides:
SMILES : CCCCCCCC(=O)OCC(COC(=O)CCCCCCC)OC(=O)CCCCCCC
CHEM : Trioctanoin
MOL FOR: C27 H50 O6
MOL WT : 470.70
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 2.45 (BCF = 281 L/kg wet-wt)
Biotransformation Half-Life (days) : 0.0857 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): 0.00 (BAF = 1 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 9.20
Log Kow (experimental): not available from database
Log Kow used by BCF estimates: 9.20

Equation Used to Make BCF estimate:
Log BCF = -0.49 log Kow + 7.554 + Correction

Correction(s): Value
Alkyl chains (8+ -CH2- groups) -0.596

Estimated Log BCF = 2.448 (BCF = 280.6 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 3 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.1024
Frag | 3 | Ester [-C(=O)-O-C] | -0.7605 | -2.2816
Frag | 3 | Methyl [-CH3] | 0.2451 | 0.7353
Frag | 20 | -CH2- [linear] | 0.0242 | 0.4837
Frag | 1 | -CH- [linear] | -0.1912 | -0.1912
L Kow| * | Log Kow = 9.20 (KowWin estimate) | 0.3073 | 2.8284
MolWt| * | Molecular Weight Parameter | | -1.2070
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | -1.0671
RESULT | Bio Half-Life (days) | | 0.08569
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 8.089 /day (10 gram fish)
kM (Rate Constant): 4.549 /day (100 gram fish)
kM (Rate Constant): 2.558 /day (1 kg fish)
kM (Rate Constant): 1.438 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = -0.000 (BCF = 0.9996 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 0.001 (BAF = 1.001 L/kg wet-wt)
Estimated Log BCF (mid trophic) = 0.033 (BCF = 1.078 L/kg wet-wt)
Estimated Log BAF (mid trophic) = 0.150 (BAF = 1.412 L/kg wet-wt)
Estimated Log BCF (lower trophic) = 0.042 (BCF = 1.102 L/kg wet-wt)
Estimated Log BAF (lower trophic) = 0.814 (BAF = 6.521 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = 2.680 (BCF = 478.2 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 6.374 (BAF = 2.367e+006 L/kg wet-wt)

For C18 chain length i.e. tristerain or glycerol tristearate:
SMILES : O=C(OC(COC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC)CCCCCCCCCCCCCCCCC
CHEM : Tristearin
MOL FOR: C57 H110 O6
MOL WT : 891.51
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 0.50 (BCF = 3.16 L/kg wet-wt)
Biotransformation Half-Life (days) : 1.28e+003 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): -0.05 (BAF = 0.893 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 23.94
Log Kow (experimental): not available from database
Log Kow used by BCF estimates: 23.94

Equation Used to Make BCF estimate:
Log BCF = -0.49 log Kow + 7.554 + Correction

Correction(s): Value
No Applicable Correction Factors
Minimum Log BCF of 0.50 applied when Log Kow > 7

Estimated Log BCF = 0.500 (BCF = 3.162 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 3 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.1024
Frag | 3 | Ester [-C(=O)-O-C] | -0.7605 | -2.2816
Frag | 3 | Methyl [-CH3] | 0.2451 | 0.7353
Frag | 50 | -CH2- [linear] | 0.0242 | 1.2094
Frag | 1 | -CH- [linear] | -0.1912 | -0.1912
L Kow| * | Log Kow = 23.94 (KowWin estimate) | 0.3073 | 7.3565
MolWt| * | Molecular Weight Parameter | | -2.2861
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | 3.1075
RESULT | Bio Half-Life (days) | | 1281
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 0.0005411 /day (10 gram fish)
kM (Rate Constant): 0.0003043 /day (100 gram fish)
kM (Rate Constant): 0.0001711 /day (1 kg fish)
kM (Rate Constant): 9.623e-005 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = -0.049 (BCF = 0.893 L/kg wet-wt)
Estimated Log BAF (upper trophic) = -0.049 (BAF = 0.893 L/kg wet-wt)
Estimated Log BCF (mid trophic) = -0.031 (BCF = 0.9315 L/kg wet-wt)
Estimated Log BAF (mid trophic) = -0.031 (BAF = 0.9315 L/kg wet-wt)
Estimated Log BCF (lower trophic) = -0.027 (BCF = 0.9402 L/kg wet-wt)
Estimated Log BAF (lower trophic) = -0.027 (BAF = 0.9402 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = -0.049 (BCF = 0.893 L/kg wet-wt)
Estimated Log BAF (upper trophic) = -0.049 (BAF = 0.893 L/kg wet-wt)

SMILES : O=C(OC(COC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC)CCCCCCCC=C
CCCCCCCC
CHEM : Triolein
MOL FOR: C57 H104 O6
MOL WT : 885.46
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 0.50 (BCF = 3.16 L/kg wet-wt)
Biotransformation Half-Life (days) : 2.36e+003 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): -0.05 (BAF = 0.893 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 23.29
Log Kow (experimental): not available from database
Log Kow used by BCF estimates: 23.29

Equation Used to Make BCF estimate:
Log BCF = -0.49 log Kow + 7.554 + Correction

Correction(s): Value
No Applicable Correction Factors
Minimum Log BCF of 0.50 applied when Log Kow > 7

Estimated Log BCF = 0.500 (BCF = 3.162 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 3 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.1024
Frag | 3 | Ester [-C(=O)-O-C] | -0.7605 | -2.2816
Frag | 3 | Methyl [-CH3] | 0.2451 | 0.7353
Frag | 44 | -CH2- [linear] | 0.0242 | 1.0642
Frag | 1 | -CH- [linear] | -0.1912 | -0.1912
Frag | 6 | -C=CH [alkenyl hydrogen] | 0.0988 | 0.5931
Frag | 6 | -C=CH [alkenyl hydrogen] | 0.0000 | 0.0000
L Kow| * | Log Kow = 23.29 (KowWin estimate) | 0.3073 | 7.1582
MolWt| * | Molecular Weight Parameter | | -2.2706
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | 3.3727
RESULT | Bio Half-Life (days) | | 2359
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 0.0002938 /day (10 gram fish)
kM (Rate Constant): 0.0001652 /day (100 gram fish)
kM (Rate Constant): 9.291e-005 /day (1 kg fish)
kM (Rate Constant): 5.225e-005 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = -0.049 (BCF = 0.893 L/kg wet-wt)
Estimated Log BAF (upper trophic) = -0.049 (BAF = 0.893 L/kg wet-wt)
Estimated Log BCF (mid trophic) = -0.031 (BCF = 0.9315 L/kg wet-wt)
Estimated Log BAF (mid trophic) = -0.031 (BAF = 0.9315 L/kg wet-wt)
Estimated Log BCF (lower trophic) = -0.027 (BCF = 0.9402 L/kg wet-wt)
Estimated Log BAF (lower trophic) = -0.027 (BAF = 0.9402 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = -0.049 (BCF = 0.893 L/kg wet-wt)
Estimated Log BAF (upper trophic) = -0.049 (BAF = 0.893 L/kg wet-wt)

(B) Fatty acids:
For C8 chain length i.e. octanoinc acid
SMILES : O=C(O)CCCCCCC
CHEM : Octanoic acid
MOL FOR: C8 H16 O2
MOL WT : 144.22
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 0.50 (BCF = 3.16 L/kg wet-wt)
Biotransformation Half-Life (days) : 0.684 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): 1.92 (BAF = 83.6 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 3.03
Log Kow (experimental): 3.05
Log Kow used by BCF estimates: 3.05

Equation Used to Make BCF estimate:
Log BCF = 0.50 (Ionic; Log Kow dependent)


Estimated Log BCF = 0.500 (BCF = 3.162 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 1 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.0341
Frag | 1 | Aliphatic acid [-C(=O)-OH] | 0.3803 | 0.3803
Frag | 1 | Methyl [-CH3] | 0.2451 | 0.2451
Frag | 6 | -CH2- [linear] | 0.0242 | 0.1451
L Kow| * | Log Kow = 3.05 (experimental ) | 0.3073 | 0.9374
MolWt| * | Molecular Weight Parameter | | -0.3698
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | -0.1648
RESULT | Bio Half-Life (days) | | 0.6842
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 1.013 /day (10 gram fish)
kM (Rate Constant): 0.5697 /day (100 gram fish)
kM (Rate Constant): 0.3204 /day (1 kg fish)
kM (Rate Constant): 0.1802 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = 1.922 (BCF = 83.64 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 1.922 (BAF = 83.64 L/kg wet-wt)
Estimated Log BCF (mid trophic) = 1.809 (BCF = 64.46 L/kg wet-wt)
Estimated Log BAF (mid trophic) = 1.810 (BAF = 64.49 L/kg wet-wt)
Estimated Log BCF (lower trophic) = 1.767 (BCF = 58.48 L/kg wet-wt)
Estimated Log BAF (lower trophic) = 1.768 (BAF = 58.65 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = 2.081 (BCF = 120.4 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 2.143 (BAF = 139 L/kg wet-wt)


(B) Fatty acids: For C18 chain length i.e. stearic acid:
SMILES : O=C(O)CCCCCCCCCCCCCCCCC
CHEM : Stearic acid
MOL FOR: C18 H36 O2
MOL WT : 284.49
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 1.00 (BCF = 10 L/kg wet-wt)
Biotransformation Half-Life (days) : 20.4 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): 4.90 (BAF = 7.89e+004 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 7.94
Log Kow (experimental): 8.23
Log Kow used by BCF estimates: 8.23

Equation Used to Make BCF estimate:
Log BCF = 1.00 (Ionic; Log Kow dependent)


Estimated Log BCF = 1.000 (BCF = 10 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 1 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.0341
Frag | 1 | Aliphatic acid [-C(=O)-OH] | 0.3803 | 0.3803
Frag | 1 | Methyl [-CH3] | 0.2451 | 0.2451
Frag | 16 | -CH2- [linear] | 0.0242 | 0.3870
L Kow| * | Log Kow = 8.23 (experimental ) | 0.3073 | 2.5294
MolWt| * | Molecular Weight Parameter | | -0.7295
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | 1.3094
RESULT | Bio Half-Life (days) | | 20.39
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 0.034 /day (10 gram fish)
kM (Rate Constant): 0.01912 /day (100 gram fish)
kM (Rate Constant): 0.01075 /day (1 kg fish)
kM (Rate Constant): 0.006046 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = 2.331 (BCF = 214.3 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 4.897 (BAF = 7.893e+004 L/kg wet-wt)
Estimated Log BCF (mid trophic) = 2.473 (BCF = 297.4 L/kg wet-wt)
Estimated Log BAF (mid trophic) = 4.985 (BAF = 9.652e+004 L/kg wet-wt)
Estimated Log BCF (lower trophic) = 2.517 (BCF = 328.8 L/kg wet-wt)
Estimated Log BAF (lower trophic) = 5.040 (BAF = 1.097e+005 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = 3.411 (BCF = 2577 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 7.025 (BAF = 1.06e+007 L/kg wet-wt)

SMILES : O=C(O)CCCCCCCC=CCCCCCCCC
CHEM : Oleic acid
MOL FOR: C18 H34 O2
MOL WT : 282.47
--------------------------------- BCFBAF v3.01 --------------------------------
Summary Results:
Log BCF (regression-based estimate): 1.75 (BCF = 56.2 L/kg wet-wt)
Biotransformation Half-Life (days) : 19.2 (normalized to 10 g fish)
Log BAF (Arnot-Gobas upper trophic): 5.18 (BAF = 1.51e+005 L/kg wet-wt)

=============================
BCF (Bioconcentration Factor):
=============================
Log Kow (estimated) : 7.73
Log Kow (experimental): 7.64
Log Kow used by BCF estimates: 7.64

Equation Used to Make BCF estimate:
Log BCF = 1.75 (Ionic; Log Kow dependent)


Estimated Log BCF = 1.750 (BCF = 56.23 L/kg wet-wt)

===========================================================
Whole Body Primary Biotransformation Rate Estimate for Fish:
===========================================================
------+-----+--------------------------------------------+---------+---------
TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE
------+-----+--------------------------------------------+---------+---------
Frag | 1 | Linear C4 terminal chain [CCC-CH3] | 0.0341 | 0.0341
Frag | 1 | Aliphatic acid [-C(=O)-OH] | 0.3803 | 0.3803
Frag | 1 | Methyl [-CH3] | 0.2451 | 0.2451
Frag | 14 | -CH2- [linear] | 0.0242 | 0.3386
Frag | 2 | -C=CH [alkenyl hydrogen] | 0.0988 | 0.1977
Frag | 2 | -C=CH [alkenyl hydrogen] | 0.0000 | 0.0000
L Kow| * | Log Kow = 7.64 (experimental ) | 0.3073 | 2.3481
MolWt| * | Molecular Weight Parameter | | -0.7243
Const| * | Equation Constant | | -1.5371
============+============================================+=========+=========
RESULT | LOG Bio Half-Life (days) | | 1.2825
RESULT | Bio Half-Life (days) | | 19.17
NOTE | Bio Half-Life Normalized to 10 g fish at 15 deg C |
============+============================================+=========+=========

Biotransformation Rate Constant:
kM (Rate Constant): 0.03617 /day (10 gram fish)
kM (Rate Constant): 0.02034 /day (100 gram fish)
kM (Rate Constant): 0.01144 /day (1 kg fish)
kM (Rate Constant): 0.006431 /day (10 kg fish)

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):
Estimated Log BCF (upper trophic) = 2.839 (BCF = 690.2 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 5.179 (BAF = 1.509e+005 L/kg wet-wt)
Estimated Log BCF (mid trophic) = 2.985 (BCF = 966.1 L/kg wet-wt)
Estimated Log BAF (mid trophic) = 5.288 (BAF = 1.94e+005 L/kg wet-wt)
Estimated Log BCF (lower trophic) = 3.029 (BCF = 1070 L/kg wet-wt)
Estimated Log BAF (lower trophic) = 5.356 (BAF = 2.267e+005 L/kg wet-wt)

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):
Estimated Log BCF (upper trophic) = 3.733 (BCF = 5413 L/kg wet-wt)
Estimated Log BAF (upper trophic) = 7.157 (BAF = 1.436e+007 L/kg wet-wt)




Estimation by CAESAR v.2.1.14 (Due to space limitations, ony the results for the longest alkyl chain containing fatty acid and glyceride has been presented here; remaining estimations are in the CAESAR BCF report PDF under the "Attached background material") :
(A) Glycerides: For C18 chain length i.e. tristerain or glycerol tristearate:
Compound SMILES:
O=C(OCC(OC(=O)CCCCCCCCCCCCCCCCC)COC(=O)CCCCCCCCCCCCCCCCC)CCCCCCCCCCCCCCCCC
Experimental value [log(L/kg)]: -
Predicted BCF [log(L/kg)]: -0.07
Predicted BCF [L/kg]: 0.84
Predicted BCF from sub-model 1 (HM) [log(L/kg)]: 0
Predicted BCF from sub-model 2 (GA) [log(L/kg)]: 0.1
Predicted LogP (MLogP): 10.93
Structural alerts: Carbonyl residue (SR 02); >C=O group (PG 09)
Reliability: the predicted compound is outside the Applicability Domain of the model
Remarks: none
Compound SMILES:
O=C(OCC(OC(=O)CCCCCCCC=CCCCCCCCC)COC(=O)CCCCCCCC=CCCCCCCCC)CCCCCCCC=CCCCCCCCC
Experimental value [log(L/kg)]: -
Predicted BCF [log(L/kg)]: -0.07
Predicted BCF [L/kg]: 0.85
Predicted BCF from sub-model 1 (HM) [log(L/kg)]: 0
Predicted

(B) Fatty acids:
For C18 chain length i.e. stearic acid:
Compound SMILES: O=C(O)CCCCCCCCCCCCCCCCC
Experimental value [log(L/kg)]: -
Predicted BCF [log(L/kg)]: 1.76
Predicted BCF [L/kg]: 58
Predicted BCF from sub-model 1 (HM) [log(L/kg)]: 1.73
Predicted BCF from sub-model 2 (GA) [log(L/kg)]: 2.47
Predicted LogP (MLogP): 5.69
Structural alerts: Carbonyl residue (SR 02); COOH group (PG 01)
Reliability: the predicted compound is outside the Applicability Domain of the model
Remarks: none



TEST US EPA Predictions ((Due to space limitations, ony the results of the longest alkyl chain containing fatty acid and glyceride has been presented here; remaining estimations are in the US EPA TEST results PDF under the "Attached background material"):

A) Predicted Bioaccumulation factor for Tristearin (CAS 555-43-1) from Consensus method

Prediction results:
Bioaccumulation factor Log10: 0.57 (predicted value)
Bioaccumulation factor: 3.70

Individual Predictions:
1. Hierarchical clustering: N/A
2. Single model: N/A
3. Group contribution: N/A
4. FDA: 0.11
5. Nearest neighbor: 1.02


B) Predicted Bioaccumulation factor for Stearic acid (CAS 57-11-4) from Consensus method

Prediction results:
Bioaccumulation factor Log10: 1.08 (predicted value)
Bioaccumulation factor: 12.02

Individual Predictions (Log10):
1. Hierarchical clustering: 0.20
2. Single model: 1.20
3. Group contribution: 1.10
4. FDA: 0.73
5. Nearest neighbor: 2.18

Reported statistics:

Not applicable

Summary of the BCF estimations using different QSAR models

Substance	EPISuite BCFBAF (BCF L/kg wet-wt)	VEGA CAESAR (BCF L/kg wet-wt)	USEPA Test (BCF L/kg wet-wt)	Average (BCF L/kg wet-wt)	Range of BCF values (L/kg)
C8 fatty acid	3.16	7.00	11.00	11.00	11-17
C18 fatty acid	10.00	58.00	12.02	11.01
C18-unsatd. fatty acid (oleic acid)	56.2	72.00	16.55	16.55
C8 triglyceride	281.00	3.00	3.96	142.48	4-143
C18 triglyceride	3.16	0.84	3.70	3.70
C18-unsatd. Triglyceride (triolein)	3.16	0.85	9.34	9.34

Refer to the QPRF for reliability discussion on the BCF estimations.

Validity criteria fulfilled:

not applicable

Conclusions:

As per the R.7c guidance, since the Kow value for some of the constituents exceeded 10, the BCF estimations were conducted using more than one QSAR models. Based on these individual BCF estimations for the representative constituents via different QSAR models (i.e., BCFBAF v3.01 of EPIWEB v 4.1, BCF (CAESAR) 2.1.14 and T.E.S.T US EPA model v.4.1.), the overall estimated BCF value for the ‘soaps, stocks, vegetable oil, acidulated’ was considered to range between 4-143 L/kg.

Executive summary:

The BCF value of ‘soaps, stocks, vegetable oil, acidulated’ was estimated through the computation methods recommended in Chapter R.7a, in: Guidance on information requirements and chemical safety assessment. Since the Kow values for some of the constituents were >10, the calculations were performed using more than one QSAR programs such as BCFBAF v3.01 of EPIWEB v 4.1, BCF (CAESAR) 2.1.14 and T.E.S.T US EPA model v.4.1 as stated in table R.7.10-3 of the REACH guidance document and/or the ECHA practical guide 5. As the test substance is a UVCB/mixture of different constituents (i.e. glycerides of fatty acids and fatty acids) the BCF values were determined for representative substances of each class present >10% to represent the lower and upperlimit of the test substance. Hence, the BCF estimations were carried out for octanoic acid, stearic acid and oleic acid representing fatty acids and trioctanoin, tristearin and triolein representing glycerides of fatty acids. SMILES notations were used as the input parameters for the programs/softwares. The BCF values estimated using BCFBAF v.3.0 for the representative constituents ranged between 3.16 -56.2 L/kg for fatty acids and 3.16 -281 L/kg for the glycerides substance class. Except for the Kow value of tristearin, the estimation by this method is more or less accurate as the molecular weight and the Kow range for the different constituents of test substance were determined to be within the molecular weight and Kow range of the training set compounds. Hence, considering this and the fact that all the representative constituent class are hydrophobic with Kow 6, they were additionally modelled using the BCF (CAESAR) 2.1.14 model. This model is based on Dimitrov et al., 2005 experimental database, which is recognised to cover hydrophobic compounds. The BCF values estimated using Caesar BCF model for the representative constituents were estimated to range between 7-72 L/kg for fatty acids and 0.84-3 L/kg for glycerides. However, these estimations using BCF (CAESAR) were reported to be not very reliable. Therefore, to further, increase the confidence of the predictions, the BCF values were additionally estimated using the BCF interface of the US EPA T.E.S.T QSAR model. This model uses several methodologies (such as hierarchical clustering, single model, group contribution, FDA, nearest neighbour) to predict an overall average BCF value under the name ‘consensus method’. The respective average BCF ranges for the representative constituents were estimated to be 11 -16.55 L/kg for fatty acids and 3.7-9 L/kg for glycerides. However, the different training set substances used for the BCF estimation for the representative substances via the US EPA TEST program were not very similar and had presence of other functional groups. Overall, considering that the estimated values from the three models lie more or less in the same BCF range, an average estimated BCF value was determined to reduce the overall uncertainty or limitations of each of the models. Therefore, based on the average BCF estimations for the representative constituents, the estimated BCF value of the ‘soaps, stocks, vegetable oil, acidulated’can be considered to range between 4 -143 L/kg (US EPA, 2012; Zhao et al., 2008).

Description of key information

Based on the average BCF estimations for the major constituents via three QSAR models (i.e., BCFBAF v3.01 of EPIWEB v 4.1, BCF (CAESAR) 2.1.14 and T.E.S.T US EPA model v.4.1), the BCF value of the ‘soaps, stocks, vegetable oil, acidulated’ can be considered to range between 4-143 L/kg. This is further supported by the low BCF values determined in an in vitro bioaccumulation study conducted with tristearin and palmitic acid (Freitag, 1985) as well as the metabolic potential of fish to break down the triglycerides and fatty acids and use as source of energy (Tocher, 2003).

Key value for chemical safety assessment

BCF (aquatic species):

143 L/kg ww

Additional information

The BCF value of ‘soaps, stocks, vegetable oil, acidulated’ was estimated through the computation methods recommended in Chapter R.7a, in: Guidance on information requirements and chemical safety assessment. Since the Kow values for some of the constituents were >10, the calculations were performed using more than one QSAR programs such as BCFBAF v3.01 of EPIWEB v 4.1, BCF (CAESAR) 2.1.14 and T.E.S.T US EPA model v.4.1 as stated in table R.7.10-3 of the REACH guidance document and/or the ECHA practical guide 5. As the test substance is a UVCB/mixture of different constituents (i.e. glycerides of fatty acids and fatty acids) the BCF values were determined for representative substances of each class present >10% to represent the lower and upper limit of the test substance. Hence, the BCF estimations were carried out for octanoic acid, stearic acid and oleic acid representing fatty acids and trioctanoin, tristearin and triolein representing glycerides of fatty acids. SMILES notations were used as the input parameters for the programs/softwares. The BCF values estimated using BCFBAF v.3.0 for the representative constituents ranged between 3.16 -56.2 L/kg for fatty acids and 3.16 -281 L/kg for the glycerides substance class. Except for the Kow value of tristearin, the estimation by this method is more or less accurate as the molecular weight and the Kow range for the different constituents of test substance were determined to be within the molecular weight and Kow range of the training set compounds. Hence, considering this and the fact that all the representative constituent classes are hydrophobic with Kow 6, they were additionally modelled using the BCF (CAESAR) 2.1.14 model. This model is based on Dimitrov et al., 2005 experimental database, which is recognised to cover hydrophobic compounds. The BCF values estimated using Caesar BCF model for the representative constituents were estimated to range between 7-72 L/kg for fatty acids and 0.84-3 L/kg for glycerides. However, these estimations using BCF (CAESAR) were reported to be not very reliable. Therefore, to further increase the confidence of the predictions, the BCF values were additionally estimated using the BCF interface of the US EPA T.E.S.T QSAR model. This model uses several methodologies (such as hierarchical clustering, single model, group contribution, FDA, nearest neighbour) to predict an overall average BCF value under the name ‘consensus method’. The respective average BCF ranges for the representative constituents were estimated to be 11 -16.55 L/kg for fatty acids and 3.7-9 L/kg for glycerides. However, the different training set substances used for the BCF estimation for the representative substances via the US EPA TEST program were not very similar and had presence of other functional groups. Overall, considering that the estimated values from the three models lie more or less in the same BCF range, an average estimated BCF value was determined to reduce the overall uncertainty or limitations of each of the models. Therefore, based on the average BCF estimations for the representative constituents, the estimated BCF value of the ‘soaps, stocks, vegetable oil, acidulated’can be considered to range between 4 -143 L/kg (US EPA, 2012; Zhao et al., 2008).

Additional recommended weight of evidence (WoE) in chapter R.11 of REACH which supports the overall low bioaccumulation potential of the substance/constituents are:

- Low bioaccumulation based on short term tests with tristearin and palmitic acid: Low bioaccumulation potential has been interpreted based on BCF value of <10 and 60 observed for tristearin and palmitic acid in a short term bioaccumulation study conducted in fish following exposure for 3 days (Freitag et al., 1985).

- Slight water solubility: The water solubility of the test substance may range from 6.4 -40 mg/L at 20 ± 2 °C which is indicative that there would be relatively low concentrations of the substance in the aquatic environment.

- Ready biodegradability of the substance: The ready biodegradation potential of the substance under stringent test conditions is indicative that there would be relatively low concentrations of the substance in the aquatic environment thereby leading to low concentrations in aquatic organisms.

- Favourable mammalian toxicokinetic data: This includes low uptake potential of the longer chain glycerides and fatty acids constituents together with the general ability of the mammals to metabolise these constituents and use as source of energy via beta-oxidation.

- Metabolism potential in fish: Similar to the mammals, the triglycerides and their constituent fatty acids are known to be broken down via beta-oxidation to serve as an important source of metabolic energy for growth and reproduction in fish (Tocher, 2003).

Therefore, the above supportive WoE together with the estimated BCF values of the major constituents (ranging from 4 -143 L/kg) are both supportive of the fact that the substance will have a low potential for bioaccumulation in aquatic as well as terrestrial organisms.