N-1-naphthylaniline

Bioaccumulation: aquatic / sediment

Link to relevant study record(s)

Reference

Reference 1

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:

other: Data from published data base (performed according to national guideline)

Qualifier:

according to guideline

Guideline:

OECD Guideline 305 C (Bioaccumulation: Test for the Degree of Bioconcentration in Fish)

GLP compliance:

not specified

Radiolabelling:

not specified

Details on sampling:

- Sampling intervals/frequency for test organisms: every two weeks, n=2; control fish: before test initiation and at termination, n=2
- Sampling intervals/frequency for test medium samples: twice a week, n=1

Vehicle:

not specified

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution, 100times concentrated
- Controls: yes
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): If the test compound is not soluble enough in water, suitable solubilizers were used.

Test organisms (species):

Cyprinus carpio

Details on test organisms:

TEST ORGANISM
- Common name: Carp
- Source: Sugishima fish farm (123-2 Gunchiku Ichibancho, Yatsushiroshi, Kumamoto 866, Japan)
- Length at study initiation (lenght definition, mean, range and SD): about 10 cm
- Weight at study initiation (mean and range, SD): about 30 g
- Health status: external disinfection according to static condition for 24 h in an aqueous solution containing 50 mg/L Terramycin (Taito Pfizer) and 7 g/L sodium chloride
- Description of housing/holding area: Fish were reared according to flow-through system for 1-2 months after external disinfection. Glass tank of 100 L volume.
- Feeding during test
- Food type: Pelleted feed for carp, Japan Higo Shiryo K.K.
- Amount: corresponding to about 2% of the total body weight of test fish
- Frequency: twice a day by halves; on the day of fish sampling feed was not supplied.

ACCLIMATION
- Acclimation period: about 28 days in an acclimation tank according to flow-through system, then the fish were transferred to test tanks and reared again at the same temperature according to flow-through system for about one month
- Acclimation conditions (same as test or not): yes

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

56 d

Hardness:

The quality of the dilution water was confirmed to meet the ministerial ordinance of the Ministry of Health and Welfare (Aug 31, 1978) in total hardness.

Test temperature:

25 +/- 2°C

pH:

measured continously, not further specified

Dissolved oxygen:

6-8 mg/L

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Type (delete if not applicable): open
- Material, size, headspace, fill volume: glass tank, 100 L volume
- Aeration: no data
- Type of flow-through (e.g. peristaltic or proportional diluter): Aquatron
- Renewal rate of test solution: flow rate 200-800 mL/min
- No. of organisms per vessel: 10-20 fish per level
- No. of vessels per concentration (replicates): no data
- No. of vessels per control / vehicle control (replicates): 2

RANGE-FINDING / PRELIMINARY STUDY
Considering the result of preliminary test for 48h LC50 and analytical detection limit of the test substance, test concentrations were decided.

Nominal and measured concentrations:

0.1 and 0.01 mg/L (nominal)

Reference substance (positive control):

not specified

Lipid content:

5.4 %

Type:

BCF

Value:

>= 427 - <= 2 730

Basis:

not specified

Remarks on result:

other: Conc.in environment / dose:0.1 ppm

Key result

Type:

BCF

Value:

>= 889 - <= 2 490

Basis:

not specified

Remarks on result:

other: Conc.in environment / dose:0.01 ppm

no further information on results

Validity criteria fulfilled:

not specified

Conclusions:

BCF values were determined to be in the range of 427 to 2730. However, the lack of relevant data (e.g., BCF values related to whole fish or specirfic compartment, vehicle used) hampers a proper assessment of the reliability of the test results.

Reference 2

Endpoint:

bioaccumulation in aquatic species: fish

Type of information:

(Q)SAR

Adequacy of study:

weight of evidence

Study period:

19 Oct 2020

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:

1. SOFTWARE
EpiSuite v4.11 (US EPA, 2012)

2. MODEL (incl. version number)
BCFBAF v3.01

3. SMILES OR OTHER IDENTIFIERS USED AS INPUT FOR THE MODEL
N(c1ccccc1)c1cccc2ccccc12

4. SCIENTIFIC VALIDITY OF THE (Q)SAR MODEL

Method description
The BCFBAF Program is an update and expansion of the previous BCFWIN Program that was part of the EPI Suite version 3.20. The update pertains to estimation of Bioconcentration Factor (BCF). The BCFBAF program estimates BCF of an organic compound using the compound's log octanol-water partition coefficient (Kow). For the update, a more recent and better evaluated database of BCF values was used for both training and validation. The BCF data were re-regressed using the same methodology as in the original BCFWIN program.
 
The original estimation methodology used by the original BCFWIN program is described in a document prepared for the Environmental Protection Agency (Meylan et al., 1997). The estimation methodology was then published in journal article (Meylan et al, 1999). 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 (1999).
 
The BCFBAF method classifies a compound as either ionic or non-ionic. Ionic compounds include carboxylic acids, sulfonic acids and salts of sulfonic acids, and charged nitrogen compounds (nitrogen with a +5 valence such as quaternary ammonium compounds). All other compounds are classified as non-ionic.

1. Defined endpoint: log BCF (bioconcentration factor)

2. Unambiguos algorithm:
Non-Ionic compounds:
Log Kow  <  1.0: All compounds with a log Kow of less than 1.0 are assigned an estimated log BCF of 0.50 (same as in BCFWIN).
Log Kow  1.0  to 7.0: Log BCF  =  0.6598 Log Kow  -  0.333  +Σcorrection factors
Log Kow  > 7.0: Log BCF  =  -0.49 Log Kow  +  7.554  +Σcorrection factors
Ionic compounds:
log BCF  =  0.50    (log Kow < 5.0)
log BCF  =  0.75    (log Kow 5.0 to 6.0)
log BCF  =  1.75    (log Kow 6.0 to 7.0)
log BCF  =  1.00    (log Kow 7.0 to 9.0)
log BCF  =  0.50    (log Kow > 9.0)

3. Applicability domain: Currently, there is no universally accepted definition of model domains. The training sets of the models contain diverse molecules, so that the fragment libraries are abundant. It is however possible that a compound has functional groups or other structural features that are not represented in the training set and for which no fragment coefficients were developed. Additionally, there can be more instances of a given fragment than the maximum for all training set compounds. These points should be taken into consideration while interpreting test results.
Molecular weight limits of the training set: 68-959 g/mol
Log Kow limits: 1 (validity range of the QSAR equation) - 11.26 (maximum value in the training set).

4. Appropriate measures of goodness of fit, robustness and predictivity.
a) 1.0 < log Kow < 7.0: N = 396 compounds, correlation coefficient R2= 0.792, Q2= 0.78, standard deviation sd = 0.511, average deviation ad = 0.395.
b) log Kow > 7.0: N = 35 compounds, correlation coefficient R2= 0.634, Q2= 0.57, standard deviation sd = 0.538, average deviation ad = 0.396.
Overall statistics including all Non-Ionic and Ionic compounds: training set N = 537, R2= 0.833, sd = 0.502, ad = 0.382.
Validation set: N = 158, R2= 0.82, sd = 0.59, ad = 0.46.
For the training set, 72.7% predictions are within 0.5 log unit and 93.5% within 1.0 log unit.

5. Mechanistic interpretation if possible: Bioaccumulation of stable substances is determined by partitioning between aqueous and lipid phases. Estimating bioconcentration factors from octanol-water partition coefficients is well established and essentially valid for neutral organics of intermediate liphophilicity (0 < log Kow < 6). For higher log Kow values, a maximum range in log BCF of approx. 6-7 (log Kow between ca. 6-8) is observed, followed by a plateau or a gradual decrease. (Source: M. Müller and M. Nendza, Literature Study: Effects of Molecular Size and Lipid Solubility on Bioaccumulation Potential).

5. APPLICABILITY DOMAIN
The compound falls within the MW and logKow range of the substances in the training set. Thus, it is considered to be in the applicability domain of the model.

 

Guideline:

other: ECHA Guidance R.6

Principles of method if other than guideline:

- Software tool(s) used including version: Estimation Programs Interface Suite™ for Microsoft® Windows v 4.10. US EPA, United States Environmental Protection Agency, Washington, DC, USA
- Model(s) used: BCFBAF v3.01
- Model description: see field 'Justification for type of information' and 'Attached justification'
- Justification of QSAR prediction: see field 'Justification for type of information' and 'Attached justification'

GLP compliance:

no

Specific details on test material used for the study:

N(c1ccccc1)c1cccc2ccccc12

Test organisms (species):

other: fish

Route of exposure:

aqueous

Test type:

other: calculation

Water / sediment media type:

natural water: freshwater

Details on estimation of bioconcentration:

BASIS FOR CALCULATION OF BCF
- Estimation software: BCFBAF v3.01
- Result based on calculated log Pow of: 4.20 (experimental)

Type:

BCF

Value:

274

Basis:

whole body w.w.

Remarks on result:

other: The substance is within the applicability domain of the BCFBAF submodel: Bioconcentration factor (BCF; Meylan et al., 1997/1999).

Type:

BCF

Value:

592

Basis:

whole body w.w.

Calculation basis:

steady state

Remarks on result:

other: Upper trophic, incl. biotransformation estimates; The substance is within the applicability domain of the BCFBAF submodel: Arnot & Gobas BAF and steady-state BCF Arnot & Gobas, 2003).

Type:

BAF

Value:

595

Basis:

whole body w.w.

Remarks on result:

other: Upper trophic, incl. biotransformation estimates; The substance is within the applicability domain of the BCFBAF submodel: Arnot & Gobas BAF and steady-state BCF Arnot & Gobas, 2003).

Details on kinetic parameters:

Biotransformation half-life (days): 2.21
Biotransformation rate (kM, normalised to 10 g fish at 15 °C): -
The substance is within the applicability domain of the BCFBAF submodel: Biotransformation rate in fish (kM; Arnot et al., 2008a/b).

Summary Results:

Log BCF (regression-based estimate): 2.44 (BCF = 274 L/kg wet-wt)

Biotransformation Half-Life (days) : 2.21 (normalized to 10 g fish)

Log BAF (Arnot-Gobas upper trophic): 2.77 (BAF = 595 L/kg wet-wt)

 

Experimental BCF-kM Database Structure Match:

--------------------------------------------

Name     : 1-Naphthalenamine, N-phenyl-

CAS Num  : 000090-30-2

Log BCF  : 3.2309 (BCF = 1.7e+003 L/kg wet-wt)

BCF Data : BCF NonIonic Training Set

Log Bio HL: ---

Bio Data : ---

 

Log Kow (experimental): 4.20

Log Kow used by BCF estimates: 4.20

 

Equation Used to Make BCF estimate:

Log BCF = 0.6598 log Kow - 0.333 + Correction

 

Correction(s):                   Value

No Applicable Correction Factors

 

Estimated Log BCF = 2.438 (BCF = 274.3 L/kg wet-wt)

 

===========================================================

Whole Body Primary Biotransformation Rate Estimate for Fish:

===========================================================

------+-----+--------------------------------------------+---------+---------

TYPE | NUM | LOG BIOTRANSFORMATION FRAGMENT DESCRIPTION | COEFF | VALUE

------+-----+--------------------------------------------+---------+---------

Frag | 1 | Aromatic amine  [-NH2 or -NH-]        | -0.2890 | -0.2890

Frag | 1 | Unsubstituted phenyl group (C6H5-)       | -0.6032 | -0.6032

Frag | 12 | Aromatic-H                               | 0.2664 | 3.1965

Frag | 2 | Number of fused 6-carbon aromatic rings  | -0.5779 | -1.1557

Frag | 1 | Benzene                                  | -0.4277 | -0.4277

Frag | 1 | Naphthalene                              | 0.4320 | 0.4320

L Kow| * | Log Kow =  4.20 (experimental  )       | 0.3073 | 1.2908

MolWt| * | Molecular Weight Parameter               |        | -0.5623

Const| * | Equation Constant                        |        | -1.5058

============+============================================+=========+=========

RESULT  |       LOG Bio Half-Life (days)           |        | 0.3444

RESULT  |           Bio Half-Life (days)           |        |   2.21

NOTE    | Bio Half-Life Normalized to 10 g fish at 15 deg C  |

============+============================================+=========+=========

 

Biotransformation Rate Constant:

kM (Rate Constant): 0.3137 /day (10 gram fish)

kM (Rate Constant): 0.1764 /day (100 gram fish)

kM (Rate Constant): 0.09919 /day (1 kg fish)

kM (Rate Constant): 0.05578 /day (10 kg fish)

 

Arnot-Gobas BCF & BAF Methods (including biotransformation rate estimates):

Estimated Log BCF (upper trophic) = 2.773 (BCF = 592.5 L/kg wet-wt)

Estimated Log BAF (upper trophic) = 2.774 (BAF = 594.9 L/kg wet-wt)

Estimated Log BCF (mid trophic)  = 2.765 (BCF = 582.4 L/kg wet-wt)

Estimated Log BAF (mid trophic)  = 2.775 (BAF = 595.8 L/kg wet-wt)

Estimated Log BCF (lower trophic) = 2.750 (BCF = 562.9 L/kg wet-wt)

Estimated Log BAF (lower trophic) = 2.779 (BAF = 600.8 L/kg wet-wt)

 

Arnot-Gobas BCF & BAF Methods (assuming a biotransformation rate of zero):

Estimated Log BCF (upper trophic) = 3.206 (BCF = 1607 L/kg wet-wt)

Estimated Log BAF (upper trophic) = 3.687 (BAF = 4859 L/kg wet-wt)

 

Validity criteria fulfilled:

yes

Conclusions:

The BCF was predicted to be 274 L/kg ww (logBCF = 2.44). The BAF was predicted to be 595 L/kg ww for the upper trophic level. The Biotransformation half-life is 2.21 d (normalized to 10 g fish).

Executive summary:

The BCF and BAF of the substance was predicted with the model BCFBAF v3.01 (EpiSuite v4.11). The BCF was predicted to be 274 L/kg ww (logBCF = 2.44). The BAF was predicted to be 595 L/kg ww for the upper trophic level. The Biotransformation half-life is 2.21 d (normalized to 10 g fish). A measured logKow from the implemented database was used for this prediction. This prediction falls within the applicability domain of the model.

Reference 3

Endpoint:

bioaccumulation in aquatic species: invertebrate

Type of information:

experimental study

Adequacy of study:

weight of evidence

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

Following recommendations of US EPA

GLP compliance:

not specified

Radiolabelling:

yes

Details on sampling:

- Sampling intervals/frequency for test organisms: Daphnia were collected at 0, 3, 6, 12, 24, 36, 48 and 72 h after test start.
- Sampling intervals/frequency for test medium samples: Triplicate 1 mL water aliquots were taken at 0, 3, 6, 12, 24, 36, 48 and 72 h after test start.
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods): Radioactivity was determined by burning the samples in a Packard Tri-Carb Sample Oxidiser and counting the radioactivity in the collected 14CO2 by liquid scintillation counting.

Vehicle:

yes

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution
- Controls: no
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): no data

Test organisms (species):

Daphnia magna

Details on test organisms:

TEST ORGANISM
- Common name: Waterflea
- Source: biological supply house

Route of exposure:

aqueous

Test type:

static

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

72 h

Hardness:

160-180 mg/L as CaCO3

Test temperature:

20 +/- 2°C

pH:

no data

Dissolved oxygen:

no data

TOC:

no data

Salinity:

not relevant

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 5 gal glass jar
- Aeration: no data
- Renewal rate of test solution (frequency/flow rate): no
- No. of organisms per vessel: no data
- No. of vessels per concentration (replicates): 1

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: reconstituted fresh water
- Alkalinity: 110-120 mg/L as CaCO3
- Holding medium different from test medium: yes
- Intervals of test medium replacement: static exposure

Nominal and measured concentrations:

40 µg/L

Reference substance (positive control):

no

Type:

BCF

Value:

637

Basis:

whole body w.w.

Time of plateau:

12 h

Remarks on result:

other: Conc.in environment / dose:40 µg/L

Elimination:

yes

Parameter:

DT50

Depuration time (DT):

53 h

Details on kinetic parameters:

- Uptake rate constant (k1): 10.2/h
- Depuration (loss) rate constant (k2): 0.016/h
- Indication of bi- or multiphasic kinetics: no

Reference 4

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:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

Bioaccumulation test (10d uptake, 32d depuration) with bluegill sunfish under flowthrough conditions. Following recommendations of US EPA

GLP compliance:

not specified

Radiolabelling:

yes

Details on sampling:

- Sampling intervals/frequency for test organisms: 7 fish were sampled at 1.5, 3, 6, 12, 24, 48, 96, 144, 192 and 240 h after introducing fish. Additional fish were sampled at 240 h for determination of PNA residues. Remaining fish were transferred to clean flowing water for the depuration phase. 7 fish were sampled at 3, 6, 12, 24, 52, 96, 192, 388.5, 576 and 768 h after start of the elimination phase.
- Sampling intervals/frequency for test medium samples: Triplicate 1 mL water aliquots were taken daily from the exposure tank.
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods): Fish were killed in anesthetic, rinsed well with clean water, blotted dry, and frozen. Each frozen fish ws dissected into head portion (incl. gills), viscera, and "edible flesh" (decapitated and eviscerated carcass). Tissues were placed in a pre-weighed paper sample cup, immediately weighed and allowed to dry. Radioactivity was determined by burning the samples in a Packard Tri-Carb Sample Oxidiser and counting the radioactivity in the collected 14CO2 by liquid scintillation counting.

Vehicle:

yes

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution; water and stock solution were thoroughly mixed by a magnetic stirrer
- Controls: no
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): acetone
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): no data

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill sunfish
- Source: commercial fish hatchery
- Weight at study initiation (mean and range, SD): 5.40 g (range 2.3 - 11.5 g, SD +/- 1.88 g)
- Weight at termination (mean and range, SD):
- Feeding during test
- Food type: commercial trout feed
- Amount: no data
- Frequency: every other day

Route of exposure:

aqueous

Test type:

flow-through

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

10 d

Total depuration duration:

32 d

Hardness:

117.9 +/- 10.4 mg/L as CaCO3

Test temperature:

20 +/- 2°C

pH:

no data

Dissolved oxygen:

no data

TOC:

2.2 +/- 1.7 mg/L

Salinity:

not relevant

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: 90 L glass aquarium fitted with a side drain
- Aeration: yes
- Type of flow-through (e.g. peristaltic or proportional diluter): peristaltic or syringe pump
- Renewal rate of test solution (frequency/flow rate): 8.8 L/h (range 8.5-9 L/h) to give a 90% replacement time of about 24h
- No. of organisms per vessel: 160
- No. of vessels per concentration (replicates): 1
- Biomass loading rate: 9.5 g fish/L water at any one time and 4.1 g fish/L water passing through the aquarium in 24 h

TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through activated charcoal
- Chlorine: < 10 µg/L
- Alkalinity: 89.1 +/- 4.5 mg/L as CaCO3
- Conductance: 265 +/- 17.3 µhos/cm
- Holding medium different from test medium: yes
- Intervals of test medium replacement: flow-through

Nominal and measured concentrations:

0.03 mg/L

Reference substance (positive control):

no

Details on estimation of bioconcentration:

BASIS INFORMATION
- Measured logPow: 4.228

BASIS FOR CALCULATION OF BCF
- log BCF = 0.85 log P - 0.70 (fish muscle), Veith et al. 1979
- log BCF = 0.542 log P + 0.124 (whole fish), Neely et al. 1974

Type:

BCF

Value:

600

Basis:

whole body w.w.

Time of plateau:

10 d

Remarks on result:

other: Conc.in environment / dose:0.03 mg/L

Type:

BCF

Value:

227

Basis:

edible fraction

Time of plateau:

10 d

Remarks on result:

other: Conc.in environment / dose:0.03 mg/L

Type:

BCF

Value:

1 424

Basis:

other: viscera

Time of plateau:

10 d

Remarks on result:

other: Conc.in environment / dose:0.03 mg/L

Elimination:

yes

Parameter:

DT90

Depuration time (DT):

8 d

Details on kinetic parameters:

- Uptake rate constant (k1): 14.7/h
- Depuration (loss) rate constant (k2): 0.014/h
- Indication of bi- or multiphasic kinetics: no

Metabolites:

HPLC analysis of the methanol extracts of the fish exposed to 14C-PNA showed at least 3 metabolites. Approx. 60% of the radioactivity in the extract was present as unmetabolized PNA. Analysis by combined GLC-MS showed the presence of a compound with a parent ion mass of 252, suggesting a hydroxy derivative of PNA. It is likely that the metabolite is 4-hydroxyphenyl-1-1(4-hydroxynaphthyl)amine based on the metabolism in mammals of a structurally related aromatic amine, diphenylamine. However, isomeric structures with both hydroxy groups in the same ring or a hydroxy group in each ring of the naphthalene moiety cannot be ruled out. The product probably results of the oxidative metabolism of PNA by fish microsomal enzymes.

Results with reference substance (positive control):

not relevant

Details on results:

- Organ specific bioaccumulation: The concentration in the viscera was substantially higher than in the edible flesh.

Reported statistics:

not specified

Validity criteria fulfilled:

not applicable

Reference 5

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:

other: Acceptable, well documented publication which meets basic scientific principles

Qualifier:

no guideline followed

Principles of method if other than guideline:

Following recommendations of US EPA

GLP compliance:

not specified

Radiolabelling:

yes

Details on sampling:

- Sampling intervals/frequency for test organisms: 1 or 2 fish were taken from each tank at 7, 24, and 49 h after test start.
- Sampling intervals/frequency for test medium samples: Triplicate 1 mL water aliquots were taken from each tank at 7, 24, and 49 h after test start.
- Details on sampling and analysis of test organisms and test media samples (e.g. sample preparation, analytical methods): Fish were rinsed well, placed in plastic bags and killed by freezing. Each frozen fish was dissected into head portion (incl. gills), viscera, and "edible flesh" (decapitated and eviscerated carcass). Tissues were placed into paper sample cups, dried and weighed. Radioactivity was determined by burning the samples in a Packard Tri-Carb Sample Oxidiser and counting the radioactivity in the collected 14CO2 by liquid scintillation counting.

Vehicle:

yes

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

PREPARATION AND APPLICATION OF TEST SOLUTION (especially for difficult test substances)
- Method: stock solution
- Controls: no
- Chemical name of vehicle (organic solvent, emulsifier or dispersant): methanol
- Concentration of vehicle in test medium (stock solution and final test solution(s) at different concentrations and in control(s)): no data

Test organisms (species):

Lepomis macrochirus

Details on test organisms:

TEST ORGANISM
- Common name: Bluegill sunfish
- Source: commercial fish hatchery
- Weight at study initiation (mean and range, SD): 1.0 g

Route of exposure:

aqueous

Test type:

static

Water / sediment media type:

natural water: freshwater

Total exposure / uptake duration:

49 h

Hardness:

117.9 +/- 10.4 mg/L as CaCO3

Test temperature:

20 +/- 2°C

pH:

no data

Dissolved oxygen:

no data

TOC:

2.2 +/- 1.7 mg/L

Salinity:

not relevant

Details on test conditions:

TEST SYSTEM
- Test vessel:
- Material, size, headspace, fill volume: glass aquaria containing 15 L
- Aeration: yes
- Renewal rate of test solution (frequency/flow rate): no
- No. of organisms per vessel: 15
- No. of vessels per concentration (replicates): 2
- Biomass loading rate: 1 g fish/L water


TEST MEDIUM / WATER PARAMETERS
- Source/preparation of dilution water: Syracuse municipal water filtered and dechlorinated through activated charcoal
- Chlorine: < 10 µg/L
- Alkalinity: 89.1 +/- 4.5 mg/L as CaCO3
- Conductance: 265 +/- 17.3 µhos/cm
- Holding medium different from test medium: yes
- Intervals of test medium replacement: static exposure

Nominal and measured concentrations:

0.2 and 0.02 mg/L

Reference substance (positive control):

no

Type:

BCF

Value:

400

Basis:

other: head

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.02 mg/L

Type:

BCF

Value:

200

Basis:

edible fraction

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.02 mg/L

Type:

BCF

Value:

4 000

Basis:

other: viscera

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.02 mg/L

Type:

BCF

Value:

644

Basis:

other: head

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.2 mg/L

Type:

BCF

Value:

346

Basis:

edible fraction

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.2 mg/L

Type:

BCF

Value:

2 548

Basis:

other: viscera

Time of plateau:

48 h

Remarks on result:

other: Conc.in environment / dose:0.2 mg/L

Elimination:

yes

Parameter:

DT50

Depuration time (DT):

6 h

Description of key information

N-phenyl-1-naphthylamine is moderately bioaccumulative in fish and daphnia.

Additional information

In the first key study, the bioconcentration of N-phenyl-1-naphthylamine in Cyprinus carpio was examined according to OECD guideline 305C in a flow-through system for 56 days (MITI Database, 2002). As a result, moderate bioconcentration was observed with BCF values of 427 to 2730 (0.1 mg/L) and 889 to 2490 (0.01 mg/L), results were not further specified.

The second key study investigated uptake and elimination of 14C-N-phenyl-1-naphtylamine by bluegill sunfish in a flow-through system following recommendations of US EPA (Sikka et al. 1981). N-phenyl-1-naphtylamine was readily taken up, equilibrium was reached apparently between 8 and 10 days. At this time, bioconcentration factors were 600 for whole fish, 339 for edible flesh and 2063 for viscera. The uptake and depuration rate constants (whole fish) were 14.7/h and 0.14/h, respectively. Half of the 14C-N-phenyl-1-naphtylamine-derived radioactivity in the fish was eliminated in about 2 days after transfer to clean flowing water. HPLC analysis of fish exposed to 14C-N-phenyl-1-naphtylamine for 10 days indicated that about 50% of the radioactivity was present as N-phenyl-1-naphtylamine metabolite(s). A major metabolite in the fish extract was identified as a dihydroxy derivative of N-phenyl-1-naphtylamine.

Uptake and elimination of 14C-N-phenyl-1-naphtylamine was also studied in two static tests with Lepomis macrochirus and Daphnia magna, respectively (Sikka et al. 1981), yielding BCF values of 400/644 (head), 200/346 (edible fraction) and 4000/2548 (viscera) for Lepomis macrochirus (concentrations: 0.02 and 0.2 mg/L, respectively), and 637 for Daphnia magna, respectively. Half of the 14C-N-phenyl-1-naphtylamine derived radioactivity in Daphnia magna was eliminated in about 2 days after transfer to clean water.

QSAR calculations using the Arnot-Gobas method (including biotransformation) of BCFBAF v3.01 resulted in BCF value of 592.5 and a BAF value of 594.9 L/kg ww.