measured BCFL (BCFL_measured) (ratio of BCFL_predicted/BCFL_measured ranged from 0.2 to 4.8) (Table 2-6). However, the bioaccumulation classification of substances would differ between the BCF models (e.g., PeCB could be classified as a bioaccumulative (B) substance not a very bioaccumulative (vB) substance).
BCFL_predicted of the BINOX M (log Kow = 8.99) was overestimated for all models, which indicates that these BCF models may be applicable to organic chemical substances with a log Kow in the approximate range of 1 to 9 (Arnot and Gobas 2004).
Table 2-1 Chemical structures of test substances used for the study of the relationship between water solubility and magnitude of bioconcentration factor [BCF]
Cyclododecane (CD) Anthracene (AN) CAS number 294-62-2 CAS number 120-12-7
2-(2-hydroxy-3,5-di-tert-butylphenyl)-5-chlorobenzotriazole (CBT) CAS number 3864-99-1
2-hydroxy-4-(octyl)phenyl phenylmethanone (PMN) CAS number 1843-05-6
2,2'-methylenebis (4-methyl-6-tert-butylphenol) (MBMBP) CAS number 119-47-1
N N Cl N
HO
O OH
O(CH2)7CH3
OH OH
Cl
Cl Cl
Cl
Cl Cl
Table 2-2 Chemical structures of test substances used for the study of the relationship between BCF and biomagnification factor [BMF])
Hexachlorobenzene (HCB) 4,4'-methylenebis(2,6-di-tert-butylphenol) (Binox M) CAS number 118-74-1 CAS number 118-82-1
Pentachlorobenzne (PeCB) 2, 4-dichloro-1-(4-nitrophenoxy) benzene (NIP) CAS number 608-93-5 CAS number 1836-75-5
1-tert-butyl -3,5-dimethyl-2,4,6- trinitrobenzene (Musk-xylene) CAS number 81-15-2
1,4-bis(isopropylamino)anthraquinone (Solvent Blue 36) o- terphenyl (TP) CAS number 14233-37-5 CAS number 84-15-1
N,N'-di-2-naphthyl-p-phenylenediamine (DNPD) Methoxychlor (MXC) CAS number 93-46-9 CAS number 72-43-5
CH2 HO
C(CH3)3
C(CH3)3
OH C(CH3)3
C(CH3)3
O2N
NO2 O2N
NH
HN Cl
Cl Cl
O O
Cl
Cl Cl
Cl
Cl Cl
O Cl
Cl
NO2
O
O HN
NH CHCH3
H3CHC CH3
CH3
Table 2-3 Exposure concentration and BCFs of test substances
Substancea log Kowb Water solubility (mg L-1) (Sw)c Exposure concentration (mg/L) (Cw) BCF_measuredd (BCF_corrected)e 2.2 × 10-1 1000 (21,000)
2.3 × 10-2 5800 (13,000) 2200 (5000)
CD 6.12 1.0 × 10-2
2.6 × 10-3 9200
1.7 × 100 100 (3400) 100 (3400) 1.3 × 10-1 1600 (4200) 1800 (4700)
1.2 × 10-2 2200 2200
AN 4.35 5.0 × 10-2
1.5 × 10-3 1800
4.3 × 10-1 1.0 (13,000)
3.6 × 10-2 6.2 (6800)
9.5 × 10-4 990 (28,000)
9.3 × 10-5 5000 (14,000) 7300 (20,000)
CBT 6.91 3.3 × 10-5
9.5 × 10-6 5700
4.2 × 10-1 25 (1600)
3.1 × 10-2 42 (200)
2.0 × 10-3 170
PMN 6.96 6.4 × 10-3
2.0 × 10-4 110
9.5 × 10-1 27 (1300)
9.1 × 10-2 100 (480)
MBMBP 7.97 1.9 × 10-2
1.8 × 10-3 730
1.6 × 10-4 550
a CD, cyclododecane; AN, anthracene; CBT, 2-(2-hydroxy-3,5-di-tert-butylphenyl)-5-chlorobenzotriazole; PMN, [2-hydroxy-4-(octyl)phenyl] phenyl methanone;
MBMBP, 2,2’-methylenebis (4-methyl-6-tert-butylphenol).
b Calculated by using Kowwin v. 1.68 (US Environmental Protection Agency).
c Measured by following the OECD TG 105 (OECD 1995).
d Average BCF on the final day of exposure phase.
e BCF_corrected estimates derived from equation (4) (Arnot and Gobas 2006).
Table 2-4 Parameters used for calculation for BMFs of test substances
Binox M, 4,4'-methylenebis (2,6-di-tert -butylphenol); PeCB, pentachlorobenzene; NIP, 2,4-dichloro-1-(4-nitrophenoxy) benzene; Solvent Blue 36, 1,4-bis(isopropylamino) anthraquinone; DNPD, N,N'-di-2-naphthyl-p-phenylenediamine; Musk-xylene,
1-tert-butyl-3,5-dimethyl-2,4,6-trinitrobenzene; TP, o-terphenyl; MXC, methoxychlor; HCB, hexachlorobenzene.
Cfood, average concentration in diet; fishlipid, average lipid content in fish based on wet weight; C0,fish, concentration in fish at the beginning of depuration derived from the regression line intercept; Ktotal, overall elimination rate constants; Kgrowth, overall growth rate constants; Kelim, growth-corrected elimination rate constants; t1/2 , growth-corrected elimination half life; alpha (α), calculated uptake efficiency of test substances from diet; BMF, biomagnification factor; Lipid factor (F), lipid-corrected factor (average lipid content in diet based on wet weight
[fishlipid]/[foodlipid]); BMFL, lipid-corrected biomagnification factor.
a Single diet group, group of one test substance + reference substance (HCB).
b Complex diet group, group of three test substances + reference substance (HCB).
c Parentheses show values of the reference substance (HCB).
Musk-xylene TP MXC
Cfood (µg g–1) 95.2 (96.8) 100 (106) 104 (94.1) 98.1 (101) 114 (103) 45.1 45.2 88.2 (24.1)
fishlipid (%) 4.51 (4.51) 4.48 (4.48) 5.34 (5.34) 4.84 (4.84) 4.83 (4.83) 6.75 6.75 6.75 (6.75)
C0,fish (µg g–1) 7.64 (16.3) 9.66 (16.1) 5.38 (14.8) 7.17 (16.0) 2.35 (16.8) 4.60 1.55 1.13 (3.24)
Ktotal (d–1) 0.0472 (0.0918) 0.213 (0.0871) 0.269 (0.0846) 0.200 (0.0686) 0.255 (0.0680) 0.130 0.372 0.376 (0.0571)
Kgrowth (d–1) 0.0231 (0.0231) 0.0171 (0.0171) 0.0291 (0.0291) 0.0298 (0.0298) 0.0288 (0.0288) 0.0238 0.0238 0.0238 (0.0238)
Kelim (d–1) 0.0241 (0.0686) 0.196 (0.0700) 0.240 (0.0555) 0.170 (0.0389) 0.226 (0.0392) 0.106 0.348 0.353 (0.0333)
t1/2 (d) 28.8 (10.1) 3.5 (9.9) 2.9 (12.5) 4.1 (17.8) 3.1 (17.7) 6.5 2.0 2.0 (20.8)
alpha (α) 0.336 (0.860) 0.777 (0.760) 0.497 (0.776) 0.564 (0.728) 0.190 (0.750) 0.541 0.430 0.162 (0.488)
BMF 0.418 (0.376) 0.119 (0.326) 0.0622 (0.420) 0.0993 (0.562) 0.0252 (0.573) 0.153 0.0370 0.0138 (0.439)
Lipid factor (F) 0.293 (0.293) 0.291 (0.291) 0.347 (0.347) 0.314 (0.314) 0.314 (0.314) 0.406 0.406 0.406 (0.406)
BMFL 1.43 (1.28) 0.410 (1.12) 0.179 (1.21) 0.316 (1.79) 0.0802 (1.83) 0.377 0.0912 0.0340 (1.08)
Binox M
Single diet groupa, c Complex diet groupb, c
Parameter
DNPD Solvent Blue 36
NIP PeCB
Table 2-5 BCFs and 5% lipid normalized bioconcentration factors (BCFL) of test substances Substance log Kowa BCFb Lipid contentb BCFL
HCB 5.86 17,000 3.10 27,000
Binox M 8.99 9200 5.70 8100
PeCB 5.22 5100 3.48 7400
Musk-xylene 4.45 4300 3.13 6500
NIP 4.32 3400 3.50 4900
Solvent Blue 36 6.07 5300 4.98 5300
DNPD 6.39 1100 3.77 1500
TP 5.52 1400 6.07 1200
MXC 5.67 620 3.83 810
a Calculated by using Kowwin v. 1.68 (US Environmental Protection Agency).
b Data from the Japanese Chemical Substances Control Law (CSCL) test reports.
Table 2-6 Measured 5% lipid normalized bioconcentration factors (BCFL_measured) and predicted BCFs (BCFL_predicted)
HCB Binox M PeCB Musk-
xylene NIP Solvent
Blue 36 DNPD TP MXC
BCFL_measured 27,000 (1) 8100 (1) 7400 (1) 6900 (1) 4900 (1) 5300 (1) 1500 (1) 1200 (1) 810 (1) BCFL_predicteda
Sijm et al. (1995) 6700 (0.2) 16,000 (1.9) 1900 (0.3) 2100 (0.3) 1700 (0.3) 2300 (0.4) 1700 (1.1) 650 (0.5) 640 (0.8) Barber (2003) 7600 (0.3) 18,000 (2.3) 2100 (0.3) 2200 (0.3) 2000 (0.4) 2700 (0.5) 2000 (1.3) 680 (0.6) 670 (0.8) Hendriks et al. (2001) 10,000 (0.4) 24,000 (2.9) 2700 (0.4) 2600 (0.4) 1800 (0.4) 3400 (0.6) 2600 (1.7) 1000 (0.9) 1000 (1.3) Arnot and Gobas (2004) 16,000 (0.6) 39,000 (4.8) 4600 (0.6) 5000 (0.7) 4200 (0.8) 5600 (1.0) 4200 (2.8) 1500 (1.3) 1500 (1.9) CERI (this study)b 17,000 (0.6) 18,000 (2.2) 6200 (0.8) 5800 (0.8) 3100 (0.6) 5000 (0.9) 1600 (1.1) 1800 (1.5) 790 (1.0)
Value in parentheses shows the ratio of BCFL_predicted/BCFL_measured.
a BCFL_predicted is derived from growth-corrected elimination rate (Kelim) data in this study.
b BCFL_predicted is derived from equation (5) using BMFL values in this study.
Figure 2-1-1 Pretreatment for analysis of 4,4'-methylenebis (2,6-di-tert -butylphenol) (Binox M), 1,4-bis(isopropylamino) anthraquinone (Solvent Blue 36) and N,N'-di-2- naphthyl-p-phenylenediamine (DNPD) in the test diet.
Test diet
Supernatant Residue
・Taking out 1 g (analytical balance)
←Tetrahydrofuran 15 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (tetrahydrofuran 5 mL)
・Centrifugation (7000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 25 mL (tetrahydrofuran, volumetric flask) Sample for LC-MS/MS analysis (Binox M and DNPD)
Sample for LC-MS analysis (Solvent Blue 36)
Figure 2-1-2 Pretreatment for analysis ofpentachlorobenzene (PeCB) in the test diet.
Test diet
Supernatant Residue
・Taking out 1 g (analytical balance)
←n-Hexane 15 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (n-hexane 5 mL)
・Centrifugation (7000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 25 mL (n-hexane, volumetric flask) Sample for GC-MS analysis
Figure 2-1-3 Pretreatment for analysis of 1-tert-butyl-3,5-dimethyl-2,4,6-trinitrobenzene (Musk-xylene), 2,4-dichloro-1-(4-nitrophenoxy) benzene (NIP), o-terphenyl (TP), methoxychlor (MXC) and hexachlorobenzene (HCB) in the test diet.
Test diet
Supernatant Residue
・Taking out 1 g (analytical balance)
←Acetone 15 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (acetone 5 mL)
・Centrifugation (7000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 25 mL (acetone, volumetric flask) Sample for GC-MS analysis
Conditions of column chromatograph
Sep-Pak Plus Silica
Conditionings n-Hexane/chloroform (80/20 V/V) approx. 10mL n-Hexane approx. 10mL Loading Whole volume of the solution was loaded.
Elution The 1st eluent n-Hexane 9 mL
The 2nd eluent n-Hexane/chloroform (80/20 V/V) 10mL The loading and the 1st and 2nd elutent were collected.
Figure 2-2-1 Pretreatment for analysis of 4,4'-methylenebis (2,6-di-tert-butylphenol) (Binox M) in the test fish.
・Measurement of weight and body length
・Chopping into pieces (scissors)
・ Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 1 g (analytical balance)
←Anhydrous sodium sulfate approx. 15 g (even balance)
←Hydrochloric acid 0.1 mL (measuring pipette)
・Dehydration (ambient temperature)
←Acetonitrile 40 mL (graduated cylinder)
・Reflux (water bath, approx. 50 ºC, 60 min.)
・Cooling (in ice water)
・Shaking (approx. 10 min.)
←Acetonitrile 20 mL (graduated cylinder)
・Shaking (approx. 10 min.)
・ Filtration (No.2 paper filter)
・ Filling up to 100 mL (acetonitrile, volumetric flask)
・Taking out 8 mL (transfer pipette)
・Evaporation to dryness
(rotary evaporator, approx. 50 ºC, nitrogen purge)
←n-Hexane 1 mL (measuring pipette)
・Ultrasonic irradiation (approx. 1 min.)
・Column chromatography
・Evaporation to dryness
(rotary evaporator, approx. 40 ºC, nitrogen purge)
←Methanol containing 5 mmol L–1
ammonium acetate 1 mL (measuring pipette)
・Ultrasonic irradiation (approx. 1 min.)
・Filling up to 2 mL (Methanol containing 5 mmol L–1 ammonium acetate, volumetric flask)
・ Taking out 3 - 5 g (analytical balance) Sample for storage
Fine sample
Residue Supernatant
Sample for LC-MS/MS analysis
×2
Supernatant Residue
Eluate Test fish
Figure 2-2-2 Pretreatment for analysis of pentachlorobenzene (PeCB) in the test fish.
・Taking out 1 - 5 g (analytical balance) Sample for storage
Fine sample Test fish
Supernatant Residue
・Measurement of weight and body length
・Chopping into pieces (scissors)
・Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 1 g (analytical balance)
・Dehydration (anhydorous sodium salfate approx. 6 g, even balance)
←n-Hexane 20 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (n-hexane 3 mL)
・Centrifugation (7000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 50 mL (n-hexane, volumetric flask) Sample for GC-MS analysis
Conditions of column chromatograph Sep-Pak Plus Silica
Conditionings n-Hexane/ethyl acetate (1/1 V/V) approx. 10mL Loading Whole volume of the solution was loaded.
Elution Eluent n-Hexane/ethyl acetate (1/1 V/V) 10mL The loading and eluent were collected.
Figure 2-2-3 Pretreatment for analysis of 1-tert-butyl-3,5-dimethyl-2,4,6- trinitro- benzene (Musk-xylene), o-terphenyl (TP), methoxychlor (MXC) and hexachlorobenzene (HCB) in the test fish.
・Taking out 1 - 5 g (analytical balance) Sample for storage
Fine sample Test fish
Supernatant Residue
・Measurement of weight and body length
・Chopping into pieces (scissors)
・Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 5 g (analytical balance)
←Acetone 25 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (acetone 3 mL)
・Centrifugation (7000 g, 5 min.)
・Dewatering filtration (anhydrous sodium sulfate approx. 40 g, even balance)
・Filling up to 50 mL (acetone, volumetric flask)
・Taking out 10 mL (transfer pipette)
・Evaporation to dryness (rotary evaporator, approx. 40 ºC)
・Column chromatography
・Evaporation to dryness (rotary evaporator, approx. 40 ºC)
・Filling up to 2 mL (n-hexane, volumetric flask) Sample for GC-MS analysis
Eluate
Figure 2-2-4 Pretreatment for analysis of 2,4-dichloro-1-(4-nitrophenoxy) benzene (NIP) in the test fish.
・Taking out 1 - 5 g (analytical balance) Sample for storage
Fine sample Test fish
Supernatant Residue
・Measurement of weight and body length
・Chopping into pieces (scissors)
・Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 5 g (analytical balance)
←Acetone 15 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (acetone 5 mL)
・Centrifugation (7000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 50 mL (acetone, volumetric flask) Sample for GC-MS analysis
Conditions of column chromatograph
Sep-Pak Plus Silica
Conditionings n-Hexane approx. 10mL
Loading Whole volume of the solution was loaded.
Elution The 1st eluent n-Hexane 10 mL
The 2nd eluent n-Hexane/ethyl acetate (10/1 V/V) 10mL The 2nd eluent was collected.
Figure 2-2-5 Pretreatment for analysis of 1,4-bis(isopropylamino) anthraquinone (Solvent Blue 36) in the test fish.
・Taking out 1 - 5 g (analytical balance) Sample for storage
Fine sample Test fish
Supernatant Residue
・Measurement of weight and body length
・Chopping into pieces (scissors)
・ Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 5 g (analytical balance)
←Acetonitrile 20 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (acetonitrile 5 mL)
・ Centrifugation (7000 g, 5 min.)
・ Filtration (absorbent cotton)
・ Filling up to 50 mL (acetonitrile, volumetric flask)
・ Taking out 1 mL (transfer pipette)
・Evaporation to dryness (rotary evaporator, approx. 40 ºC, nitrogen purge)
←n-Hexane 1 mL (measuring pipette)
・Column chromatography
・Evaporation to dryness (rotary evaporator, approx. 40 ºC, nitrogen purge)
←Tetrahydrofuran 4 mL (measuring pipette)
←Ultra pure water 4 mL (measuring pipette)
・Filling up to 10 mL (tetrahydrofuran/ultra pure water (1/1 V/V), volumetric flask)
Sample for LC-MS analysis Eluate
Conditions of column chromatograph Sep-Pak Plus Silica
Conditionings Tetrahydrofuran approx. 10mL
Loading Whole volume of the solution was loaded.
Elution Eluent Tetrahydrofuran 4mL
The loading and eluent were collected.
Figure 2-2-6 Pretreatment for analysis of N,N'-di-2-naphthyl-p-phenylenediamine (DNPD) in the test fish.
・Taking out 1 - 5 g (analytical balance) Sample for storage
Fine sample Test fish
Supernatant Residue
・Measurement of weight and body length
・Chopping into pieces (scissors)
・Refinement (Polytron, 2 min. or more, on ice water)
・Taking out 5 g (analytical balance)
←Tetrahydrofuran 30 mL (graduated cylinder)
・Homogenization (Polytron, approx. 1 min.)
・Washing (tetrahydrofuran 5 mL)
・Centrifugation (4000 g, 5 min.)
・Filtration (absorbent cotton)
・Filling up to 50 mL (tetrahydrofuran, volumetric flask)
・Taking out 1 mL (transfer pipette)
・Column chromatography
・Filling up to 10 mL (ultra pure water, volumetric flask) Sample for LC-MS/MS analysis
Eluate
Figure 2-3 Relationship between exposure concentration and bioconcentration factor (BCF) of test substances.
CD, cyclododecane; AN, anthracene; CBT, 2-(2-hydroxy-3,5-di-tert-butylphenyl)-5-chlorobenzotriazole;
PMN, [2-hydroxy-4-(octyl)phenyl] phenyl methanone; MBMBP, 2,2'-methylenebis (4-methyl-6-tert-butylphenol).
Black diamonds show measured BCF and red diamonds show corrected BCF derived from equation (4) (Arnot and Gobas 2006).
Solid line shows the measured water solubility following the OECD TG 105 (OECD 1995).
2 3 4 5
10-3 10-2 10-1 100
Exposure concentration (mg L-1)
log BCF
(a) CD
1 2 3 4 5
10-3 101
Exposure concentration (mg L-1)
log BCF
10-2 10-1 100 (b) AN
0 2 4 6
Exposure concentration (mg L-1)
log BCF
10-5 10-4 10-3 10-2 10-1 100 (c) CBT
1 2 3 4
Exposure concentration (mg L-1)
log BCF
(d) PMN
10-4 10-3 10-2 10-1 100
(e) MBMBP
1 2 3 4
Exposure concentration (mg L-1)
log BCF
10-4 10-3 10-2 10-1 100 2
3 4 5
10-3 10-2 10-1 100
Exposure concentration (mg L-1)
log BCF
(a) CD
1 2 3 4 5
10-3 101
Exposure concentration (mg L-1)
log BCF
10-2 10-1 100 (b) AN
0 2 4 6
Exposure concentration (mg L-1)
log BCF
10-5 10-4 10-3 10-2 10-1 100 (c) CBT
1 2 3 4
Exposure concentration (mg L-1)
log BCF
(d) PMN
10-4 10-3 10-2 10-1 100
(e) MBMBP 2
3 4 5
10-3 10-2 10-1 100
Exposure concentration (mg L-1)
log BCF
(a) CD
2 3 4 5
10-3 10-2 10-1 100
Exposure concentration (mg L-1)
log BCF
(a) CD
1 2 3 4 5
10-3 101
Exposure concentration (mg L-1)
log BCF
10-2 10-1 100 (b) AN
1 2 3 4 5
10-3 101
Exposure concentration (mg L-1)
log BCF
10-2 10-1 100 (b) AN
0 2 4 6
Exposure concentration (mg L-1)
log BCF
10-5 10-4 10-3 10-2 10-1 100 (c) CBT
0 2 4 6
Exposure concentration (mg L-1)
log BCF
10-5 10-4 10-3 10-2 10-1 100 (c) CBT
1 2 3 4
Exposure concentration (mg L-1)
log BCF
(d) PMN
10-4 10-3 10-2 10-1 100 1
2 3 4
Exposure concentration (mg L-1)
log BCF
(d) PMN
10-4 10-3 10-2 10-1 100
(e) MBMBP
1 2 3 4
Exposure concentration (mg L-1)
log BCF
10-4 10-3 10-2 10-1 100 1
2 3 4
Exposure concentration (mg L-1)
log BCF
10-4 10-3 10-2 10-1 100
Figure 2-4-1 Concentration of 4,4'-methylenebis(2,6-di-tert-butylphenol) (Binox M) and hexachlorobenzene (HCB) concentration in test fish during test period.
Blue (Binox M) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <10 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >10 0
10 20
0 5 10 15 20 25 30 35 40 45 50
Test period (d) Concentration in test fish (µg g-1) a
0 10 20
0 5 10 15 20 25 30 35 40 45 50
Test period (d) Concentration in test fish (µg g-1) a Uptake phase Depuration phase
Figure 2-4-2 Concentration of pentachlorobenzne (PeCB) and hexachlorobenzene (HCB) concentration in test fish during test period.
Blue (PeCB) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <10 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >10 0
10 20
0 5 10 15 20 25 30 35 40
Test period (d) Concentration in test fish (µg g-1) a
0 10 20
0 5 10 15 20 25 30 35 40
Test period (d) Concentration in test fish (µg g-1) a
Uptake phase Depuration phase
Figure 2-4-3 Concentration of 2,4-dichloro-1-(4-nitrophenoxy) benzene (NIP) and hexachlorobenzene (HCB) concentration in test fish during test period.
Blue (NIP) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <10 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >10
Uptake phase Depuration phase
0 10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish (µg g-1) a
0 10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish (µg g-1) a
0 10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish (µg g-1) a
0 10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish (µg g-1) a
Figure 2-4-4 Concentration of 1,4-bis(isopropylamino)anthraquinone (Solvent Blue 36) and hexachlorobenzene (HCB) concentration in test fish during test period.
Blue (Solvent Blue 36) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <10 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >10
Uptake phase Depuration phase
Figure 2-4-5 Concentration of N,N'-di-2-naphthyl-p-phenylenediamine (DNPD) and hexachlorobenzene (HCB) concentration in test fish during test period.
Blue (DNPD) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <10 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >10 0
10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish µg g-1) a
0 10 20
0 5 10 15 20 25 30
Test period (d) Concentration in test fish µg g-1) a Uptake phase Depuration phase
Figure 2-4-6 Concentration of 1-tert-butyl -3,5-dimethyl-2,4,6- trinitrobenzene (Musk-xylene), o- terphenyl (TP), methoxychlor (MXC) and hexachlorobenzene (HCB) concentration in test fish during test period.
Purple (Musk-xylene), green (TP), blue (MXC) and red (HCB) diamond shows measured concentration in test fish.
Solid line shows the calculated concentration in test fish using following equation.
Cfish = Cdiet × I×α × (1-e-ktotal × t) / ktotal 0 < t <13 Cfish = Cdiet × I × α × (e-ktotal × (t-10) - e-ktotal × t) / ktotal t >13 0
5 10
0 5 10 15 20 25 30 35
Test period (d) Concentration in test fish (µg g-1) a
0 5 10
0 5 10 15 20 25 30 35
Test period (d) Concentration in test fish (µg g-1) a
Uptake phase Depuration phase
Figure 2-5 Relationship between 5% lipid normalized bioconcentration factor (BCFL) and lipid corrected biomagnification factor (BMFL) of test substances.
Solid line (log [BCFL] = 0.828 log [BMFL] + 4.12, r2 = 0.873) shows the linear regression and dashed lines show 95% confidence interval.
BMFL BCFL
102 103 104 105
10-2 10-1 100 101
MXC TP
DNPD
NIP Solvent Blue 36
Musk-xylenePeCB
Binox M HCB
Figure 2-6 The correlation between bioconcentration factor (BCF) and bioaccumulation factor (BAF) data of existing persistent organic pollutants (POPs) and POP candidates.
The figure including BCF and BAF data are from Kitano (2007). Blue diamonds show BCF data and red circles show BAF data. Orange lines show BCFL = 5600 and 30,000 (range of BMFL = 1) derived from equation (5) (incorporating 95% confidence interval). Information for the POPs and POP candidates is available at http://chm.pops.int/default.aspx
0 1 2 3 4 5 6 7 8
PeCB SCCP OcBDE beta-HCH alpha-HCH Lindane Chlordecone HeBB PFOS PeBDE PCDFs PCDDs PCBs Mirex Heptachlor HCB DDT Chlordane Endrin Dieldrin
Aldrin BCFL < 5600 (BMFL < 1) Less likely to biomagnify via food chain
BCFL > 5600 (BMFL > 1) Likely to biomagnify via food chain
log BCF or log BAF
0 1 2 3 4 5 6 7 8
PeCB SCCP OcBDE beta-HCH alpha-HCH Lindane Chlordecone HeBB PFOS PeBDE PCDFs PCDDs PCBs Mirex Heptachlor HCB DDT Chlordane Endrin Dieldrin
Aldrin BCFL < 5600 (BMFL < 1) Less likely to biomagnify via food chain
BCFL > 5600 (BMFL > 1) Likely to biomagnify via food chain
log BCF or log BAF
Chapter 3
Comparison of Nitrofen uptake via water and food and its distribution in tissue of common carp (Cyprinus carpio)
1. Introduction
In Chapter 2, the author showed a good relationship between bioconcentration factor (BCF) and biomagnification factor (BMF) of poorly water-soluble chemicals and explored the effectiveness of these bioaccumulation endpoints.
In general, BCF and BMF are calculated using the concentration of chemical substance in an organism on whole body basis. In CSCL (2003), if BCF is between 1000 and 5000, BCF of each fish part (especially edible parts, i.e., fish muscle) should be considered to evaluate the bioaccumulation potential of chemical substance, because fish muscle is a primary source of protein for human consumption, especially Japanese.
However, there is little experimental data that compares the tissue distribution of chemical substances taken up by different routes (e.g., via water or food). Nitrofen (1og Octanol-Water Partition Coefficient 4.32, calculated by using Kowwin v. 1.68 [US Environmental Protection Agency]) is highly hazardous to aquatic organisms; however there is limited hazard data (Hazardous Substances Data Bank;
http://toxnet.nlm.nih.gov). Therefore, in Chapter 3, the author investigates the uptake characteristics of Nitrofen in carp (Cyprinus carpio) via water and food and discusses its tissue distribution on the basis of lipid content.
2. Materials and Methods