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Bioaccumulation: aquatic / sediment

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Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Study completion date - 06 June 1973.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
guideline study with acceptable restrictions
Justification for type of information:
Read-across based on grouping of substances (category approach). A detailed category justification is included in chapter 13 of this dossier.
Principles of method if other than guideline:
Bluegill sunfish (Lepomis macrochirus) were exposed for up to 70 days in dynamic tank water systems containing the test substance at concentrations of 1.0, 0.01, 0.001 and 0 mg/l, respectively. It was necessary to repeat a portion of the original study; the second study was carried out both to establish the accumulation of test item in fish in the absence of disease and to confirm that the 56 day samples from the original study were indeed contaminated and did not represent some unusual accumulation pattern on the part of the fish (BASF, 1972).
GLP compliance:
no
Remarks:
Pre GLP.
Details on sampling:
- Sampling frequency for test organisms: at day 1, 3, 7, 14, 28, 42, 56, 70.
- No of organisms sampled: at each sampling interval, three (3) bluegill were removed from each experimental unit (including Blank unit).
- Sampling during the withdrawal phase: taken at 0, 1, 3, 7 and 14 day intervals.
- Sample storage conditions before analysis: all samples were frozen.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
- Average weight at study initiation: 50 g.
Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
70 d
Details on test conditions:
TEST SYSTEM
- Test vessel: 150 gallon fibre glass tank.
- Type of flow-through: five direct dilution metering systems.
- Renewal rate of test solution: 2-3 minutes.
- No. of organisms per vessel: 40-50.

TEST MEDIUM
An investigation of the suitability of fibre glass tanks as the test units was conducted prior to starting the definitive study. The system was ran at the desired concentration (without fish) for 7 days during which time water samples were removed after 1, 3 and 7 day exposure, and sent to CIBA-GEIGY for analysis. Duplicate, samples were sent in glass and metal containers to evaluate the suitability of metal containers for shipping water samples. The tank vessels then were "cleaned" and the system ran an additional 7 days without chemical. Water samples were taken after 1, 3 and 7 days and sent to CIBA-GEIGY for analysis to evaluate the amount. If any, of leaching of chemical residues from fibre glass tanks into the water.

WITHDRAWAL PHASE
When analytical results show that no significant increase in tissue residues occurred with increasing exposure time, the exposure phase of the study was terminated and the withdrawal phase begun.
Nominal and measured concentrations:
Nominal: 0.001, 0.01, 0.1 and 1 mg/l. Analysis of the water samples showed that with the exception of the one and three-day sampling periods the concentration of the test substance in the test tanks was usually within ±15 % of the nominal levels.
Details on results:
The data from the two studies confirmed that with the exception of the contaminated 56 day samples, the concentration of test item found in the edible portion of the exposed fish is extremely low (<0.05 mg/l) even at the 1 mg/l exposure level. From these results it appears that test substance does not accumulate in fish tissue.

It was necessary to repeat a portion of the original study; this was prompted by two factors which cast some doubt on the validity of the original study (see the key study BASF, 1972). In the first place the original study was complicated by a disease which developed in two of the test tanks (0.1 and 0.001 mg/l) during the study. Secondly the 56 day fish samples from the 0.01 and 1 mg/l tanks were contaminated with FWA from an unknown source. The second study was carried out both to establish the accumulation of test item in fish in the absence of disease and to confirm that the 56 day samples from the original study were indeed contaminated and did not represent some unusual accumulation pattern on the part of the fish (see the key study BASF, 1972).

Concentration of test item in eviscerated Bluegill sunfish (Lepomis Macrochirus) exposed to the whitener at various tank water concentrations.

Test item concentration in fish tissueab (mg/kg)
Day/phase Fish 0.001 mg/l 0.01 mg/l 0.1 mg/l 1 mg/l
Accumulation
1 A ND tr tr tr
B tr tr tr tr
3 A tr ND tr tr
B tr tr tr 0.04
7 A ND tr tr 0.04
B tr tr 0.03 0.03
C NA NA NA NA
14 A ND ND ND tr
B tr ND 0.03 ND
C NA NA NA tr
28 A NDf tr trd tr
B ND tr trd trd
C NA NA NA NA
42 A (c) ND tr 0.03
B - ND tr tr
C - NA NA NA
56 A - > 0.06g (c) 2.0f
B - 0.22f - 2.1g
C - NA - NA
70 A - (e) - (e)
B - - - -
C - - - -
Withdrawal
1 A - trfg - trfg
B - trfg - trg
C - trf - trf
3 A - NDfg - trfg
B - NDfg - trf
C - NDf - -
7 A - NDf - NDf
B - ND - -
C - - - -
14 A - NDf - NDf
B - - - -
C - - - -

a = each result is an average of duplicate analyses unless otherwise specified.

b = control samples were analyzed and found In all cases to contain no detectable whitener.

c = fish in these tanks succumbed to disease; therefore these tanks were discontinued.

d = due to small fish size or due to the necessity to dilute unusually viscous analytical solutions, the trace level was <0.04 rather than <0.03 mg/kg.

e = the duration of the accumulation phase from the initial study was 56 days; the repeat study was carried through 70 days.

f = only single determination possible due to small fish size.

g = samples received thawed.

ND = none detected (Sensitivity limit = 0.03 mg/kg).

t r = trace; detectable but below the quantizable limit.

NA = samples received but not analyzed.

NS = no samples taken.

Concentration of test item in tank water samples removed during the exposure phase

  Test item concentrationab
Day/phase 0.001 mg/l 0.01 mg/l 0.1 mg/l 1 mg/l
1 0.046 0.019 0.180 1.85
3 0.016 0.027 0.016 0.45
7 0.002 0.010 0.100 1.00
14 0.001 0.012 0.093 1.00
28 0.001 0.012 0.090 0.93
42 (c) 0.011 0.087 0.89
56 - 0.012 (c) 0.80
70 - (d) - (d)

a = Average of duplicate analyses unless otherwise specified.

b = Water was removed from the control tank on each sampling day; analyses showed no detectable Tinopal AMS in any of these samples.

c = Fish in these tanks succumbed to disease; therefore these tanks had to be discontinued.

d = The exposure phase of the original study lasted only 56 days.

Validity criteria fulfilled:
not specified
Conclusions:
Test substance does not accumulate in fish tissue.
Executive summary:

Bluegill sunfish (Lepomis macrochirus) were exposed for up to 70 days in dynamic tank water systems containing the test substance at concentrations of 1.0, 0.01, 0.001 and 0 (control) mg/l. Following the exposure phase, the remaining fish were removed to tanks containing water free of the compound to determine the rate of elimination of any accumulated whitener. Fish and water were sampled at designated intervals during the exposure and elimination phases and sent to Ciba-Geigy for analysis. It was necessary to repeat a portion of the original study; this was prompted by two factors which cast some doubt on the validity of the original study. In the first place the original study was complicated by a disease which developed in two of the test tanks (0.1 and 0.001 mg/l) during the study. Secondly the 56 day fish samples from the 0.01 and 1 mg/l tanks were contaminated with FWA from an unknown source. The second study was carried out both to establish the accumulation of test item in fish in the absence of disease and to confirm that the 56 day samples from the original study were indeed contaminated and did not represent some unusual accumulation pattern on the part of the fish (see the key study Ciba-Geigy, 1972)

Results

With the exception of the contaminated 56 day samples, the concentration of test item found in the edible portion of the exposed fish is extremely low (<0.05 mg/l) even at the 1 mg/l exposure level. From these results it appears that test substance does not accumulate in fish tissue.

Endpoint:
bioaccumulation in aquatic species: fish
Type of information:
experimental study
Adequacy of study:
supporting study
Study period:
Study completion date - 21 September 1972.
Reliability:
2 (reliable with restrictions)
Rationale for reliability incl. deficiencies:
comparable to guideline study with acceptable restrictions
Justification for type of information:
Read-across based on grouping of substances (category approach). A detailed category justification is included in chapter 13 of this dossier.
Principles of method if other than guideline:
Bluegills (Lepomis machrochirus) have been exposed in dynamic flowthrough tankwater systems to test item to determine to what degree this product accumulate in portions of the fish which may be consumed by humans. Test concentration was 1 mg/l. The exposure period was 70 days and the elimination phase was 14 days. During the tests, fish and water were periodically sampled and analysed.
GLP compliance:
no
Details on sampling:
- Tissue: The edible portion, i.e. the entire carcass of the fish with the exception of the organs contained in the visceral cavity, was analysed.
- Sample preparation: Fish were rinsed with water, rinsed with acetone, eviscerated and frozen for storage prior to analysis.
- Sampling frequency for test organisms: Bluegill from were sampled on days 1, 3, 7, 14, 21, 28, 56 and 70 of exposure.
- Sampling frequency for test medium samples: Water samples were taken before commencing fish exposure and analysed to insure that the system operated properly.
- Sampling frequency for organisms in uncontaminated water: At the termination of exposure were sampled on days 1, 3 and 7.
- N. organisms sampled: At each sampling interval (both exposure and withdrawal) five (5) fish were sampled from each unit.
Details on preparation of test solutions, spiked fish food or sediment:
PREPARATION AND APPLICATION OF TEST SOLUTION
Stock solutions were made fresh daily and shielded from exposure to light by wrapping with aluminium foil. All stock solutions were made in distilled water.
Test organisms (species):
Lepomis macrochirus
Details on test organisms:
TEST ORGANISM
- Common name: Bluegill sunfish.
- Source: A commercial fish farmer in Rhode Island.
- Average weight at study initiation: mean of 50 g.
- Length at study initiation: average of 140 mm.
- Food type: fish were fed a dry pelleted ration ad libitum each day.

ACCLIMATION
- Acclimation period: the fish were held in the hatchery facilities at the test laboratory for a minimum of 30 days prior to the initiation of the study.
- Health during acclimation: during this period the cumulative mortality was less than 1% and the fish appeared to be in excellent physical condition.
Route of exposure:
aqueous
Test type:
flow-through
Water / sediment media type:
natural water: freshwater
Total exposure / uptake duration:
70 d
Total depuration duration:
14 d
Hardness:
40 mg/l as CaCO3
Test temperature:
18 ± 0.5 °C
pH:
7.1-7.3
Dissolved oxygen:
7.5 mg/l
Details on test conditions:
TEST SYSTEM
- Test vessel: 500 litre fibre glass tanks.
- Aeration: Aerated well water.
- Type of flow-through: Modification of a continuous-flow proportional dilution apparatus which provides for the automatic intermittent introduction the test material and diluent water in the test tank.
- Renewal rate of test solution: Aerated well water was provided to each unit at a flow rate of 6 l/hour.
- No. of organisms per vessel: 60 bluegill were placed into each tank.

TEST MEDIUM / WATER PARAMETERS
- Medium control: Duplicate water samples for chemical analysis were removed periodically from each experimental unit, placed in amber glass bottles and preserved by addition of NH4OH.

EXPOSURE
- Exposure: The exposure was continued until no significant increase was noted in the concentration of test item in the fish at three successive sampling periods. The minimum exposure period was 30 days.
- Post exposure: Following the exposure phase, the remaining fish were transferred to water free of the test substance. The elimination phase was continued until no significant amounts of test item were found in the fish.
Nominal and measured concentrations:
1 mg/l nominal.
Details on results:
Only sporadic trace concentrations (less than 0.05 mg/kg) of the test item were present in the fish at exposure levels of 1 mg/l.
In most instances, no detectable substance was found. The data indicates that there is no tendency for the substance to be taken up and stored in the fish.

Concentration of test item in eviscerated Bluegill sunfish (Lepomis Macrochirus) exposed 1 mg/l.

Accumulation
Day/phase 1 3 7 14 28 56 70
Fish A B A B A B C A B C A B C A B C A B C

Test item concentration in fish tissueab (mg/kg)

NS NS NS NS tr tr tr 0.04 0.05 0.04 tr tr tr tr tr trd tr tr tr
Test item concentration in tankwater (mean) 0.85 0.93 0.95 1.10 0.87c 0.80 0.93

Withdrawal
Day/phase 1 3 7 14
Fish A B C A B C A B C A B C

Test item concentration in fish tissueab

tr tr tr ND tr tr tr tr tr NS NS NS
Test item concentration in tankwater (mean)e NS NS NS NS

a = each result is an average of duplicate analyses unless otherwise specified.

b = control samples were analyzed and found In all cases to contain no detectable whitener.

c = only single determination possible due to small fish size.

d = only single determination possible due to small fish size.

e = concentration of test substance in tank water samples removed during the exposure phase.

tr = trace; detectable but below the quantizable limit.

NS = Not sampled.

Validity criteria fulfilled:
not specified
Conclusions:
No tendency for the substance to be taken up and stored in the fish.
Executive summary:

Method

Bluegills (Lepomis machrochirus) have been exposed in dynamic flowthrough tankwater systems to test item to determine to what degree this product accumulate in portions of the fish which may be consumed by humans. The test concentration was 1 mg/l. The exposure period was 70 days and the elimination phase was 14 days. During the tests, fish and water were periodically sampled and analysed.

Results

Only sporadic trace concentrations (less than 0.05 mg/kg) of the test item were present in the fish at exposure levels of 1 mg/l. In most instances, no detectable substance was found. The data indicates that there is no tendency for the substance to be taken up and stored in the fish.

Description of key information

No bioaccumulation is expected

Key value for chemical safety assessment

BCF (aquatic species):
0.5 dimensionless

Additional information

The substance has a low potential for bioaccumulation (log Kow < 3) and a low potential to cross biological membranes (high molecular weight, high polarity); furthermore, direct and indirect exposure of the aquatic compartment is unsuitable. Nevertheless, bioaccumulation studies are available on the analogous substance of the category of Stilbene Fluorescent Whitening Agents, CAS 16090-02-1 and are here reported for completeness sake and in order to have a better overview of the PBT assessment. In all the experiments performed only sporadic trace concentrations (less than 0.05 mg/kg) of the substance were present in the fish at exposure levels up to of 1 mg/l. In most instances, no detectable substance was found. The substance CAS 16090-02-1 is the analogous substance morpholino derivative, disodium salt. This substance is much less soluble than CAS 41098-56-0 (1.9 g/l versus 201 g/l) and the morpholino group make it potentially more reactive (enzymatic opening of morpholino ring on N atom), therefore it can be considered a conservative representative for CAS 41098-56-0 regarding bioaccumulation potential. Furthermore, the Bioconcentration Factor (BCF) value was calculated using BCFBAF v3.01 tool, an application contained in the EpiSuite 4.1 [1]. The estimation takes into account the substance is a ionic compound. The BCF estimated value from regression-based method is 0.5 (BCF = 3.175 l/kg wet-wt), in agreement with the common value of non bioaccumulative substances.

[1] EpiSuite is the suite of physical/chemical property and environmental fate estimation programs developed by the EPA’s Office of Pollution Prevention Toxics and Syracuse Research Corporation (SRC).