Document 3Jj43pDjkdwpbyBBpNZ9yqpNJ
AR 226 3300
482
AR 226 _ 3300
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Study Report
Study A icn in Carp
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`September 22, 2003
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Chemicals Evaluation and Research Institute, Japan
Kurume Laboratory--
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Company Sanitized. Does not contain TSA cay
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Figures
Fig. 1
Mass fragmentograms of test water).
of
LC-MS
analysis
for
recovery
and
blank
tests
(analysis
Fig. 2 oMfatsesstffrisahg)m.entograms of LC-MS analysis for recovery and blank tests (analysis
Fig.3 Calibration curves
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Study No. 102
company Sanitized. Does notcontainTse oy
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Title:
study of{IAccomulation in Carp
Commissioned by: DuPont KK 1-8-1 Shimo-meguro, Meguro-ku, Tokyo 153-0064
Testing Facility:
Kurume Laboratory Chemicals Evaluation and Research Institute, Japan (CERI) 19-14 Chuo-machi, Kurume-shi, Fukuoka Prefecture. 830-0023
Purposoef Study: To obtain findings on the accumulation of(NDin carp.
Personnel:
ReSsteuadrycChoeorrsd:inator: YNoasohaiktiakYaAKGAOTGAA," Testing Section 2 Masanori WATASHIMA
Approval of Study Report: [signed] Naoaki YAGATA Study Coordinator `September 22, 2003
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* Translaor's Note: Spellings ofpersonal names may need to be verified.
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Study No. 44102
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1 Test
Name:
Lot No. IC] ql
py [
2. PreparationofStock Solution by first Agrisntdoicnkgsotlougteitohnerhianvianmgoarttaesrttshuebtsetsatnscuebcsotnacnecnetraantdioan2o0f-fSo0ld0 ammgo/uLnwtaosfpcrreysptaarled sugar, subsequently adding a 20-fold amount (relative to the test substance)ofMegafac F-142D and again grinding the ingredients together in a mortar, then adding ionexchanged water.
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33..1PrTeelsitmCionnadriytiToensst of Acute Toxicity
((21)) NTeusmtbfeisrh:of test fish: 2Hifmisehdapekra'co(nOcreynztiraatsiloantigpreso)up
(3) Period of exposure: 96 hours
(4) Method of exposure: semi-stagnant water method (change of water every 810 16
hours)
(5) Preliminary test results: preliminary LCso over 96 hours was >30.0 mg/L
~~ "TN: Japanese name of fish. English name i not known.
Sy No-dd102
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4. Investigation of Analysis Conditions
sensitiviBtaysoefdaonnaltyshiesp,rtehleimtiesntarcyonLcCesntorvaatliuoensowfetrhee tseesttassubfsoltlaonwcse over96 hours and the.
First concentration group: 50 ug/L
Second concentration group: 5 ug/L,
`and an examination of the analysis conditions was carried out.
4.1 Test Substance Analysis in Test Water and Test Fish `When the test substance was analyzed by liquid chromatography-mass
spectrometry (LC/MS), three peaks were detected. Quantitative determinations were thus carried out on cachofthe peaks. The respective peak concentrationsare indicated as the test substance concentrations shown in section 4.2.2 ("Preparation of Standard Solution"), regardless of the constituent compositions. The peaks were named Peak 1, Peak 2 and Peak 3, in the orofdeluetiorn.
4.1.1 Investigation of Method for Pretreating Test Water Analysis Specimens Stock solution (containin2g ug of the test substance) was added to 400 mL of test
`water for the recovery test, and pretreatment operations were carried out according to the flow scheme shown below to give a liquid chromatography-mass spectrometry (LC-MS) specimen.
Flow Scheme
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[reermini ied]
Cdeotlaiulms)n chvomatography" (sc next page for
| | VBIrNi)n.g muepa0sur1i0nmgLfl(asmke)thanol water" (872
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TCS ump
Tpiapkeettael)iquot sampleof2.5 mL. (ransfer
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Goakd)
BVrIiVn)g, umepa0sur1i0nmgLfl(asmke)thanol water (872
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eo.)
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Study No.44102
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*1: Column chromatography conditions:
SepPack Cis
Lo
(Cleaning method:
Loading method:
10mL
Entire
each of
amount
methanol */water"*
was loaded. _
(8
/2
V
/
V
)
a
nd
water
")
Elution method: First eluting solvent: water
SmL
`STehceosnedceolnudtienlgutsionlgvesnotl:vemnettwhaasnoflur"nwiastheedrfor a(n8a2lyVsi/sV.) ~~ 8 mL
#2: Contains 10mmol/L of ammonium acetate
#3: Water obtained by treating tap water using an ultrapure water production system.
`The
percent recovery in eachofthe Percent recovery (n=1)
above Peak
operations are given I: 85.1%
below
(see
Fig.
1).
Peak2 90.5%
Peak: 742%
Blank (n=1)
Peakl: notdetected
Peak2: not detected
Peak3: notdetected
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Study No. 44102
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4.1.2
InvestigationofMethod for Pretreating Stock solution (containin5g pgofthe
Test Fish Analysis Specimens test substance) was added to 5
g
ofa
finely
c`agrrroiuendd osuatmpalcecoorfditnhge tteostthfeisfhlfoowrstchheermeecosvheorywntesbte,laonwdtporegtirveeaatmleinqtuiodpcerhartoimoantsowgrearpehy-
`mass spectrometry (LC-MS) specimen.
Flow Scheme
Ee --
=
MHoemtohgaennio?zl,e 1(5PomlLytr(onm,eaasbuoruitng1 mciylniuntdee)r)
Wash (methanol, 3 mL)
Centrifugal separation (700xg, 5 minutes)
Filtration (absorbent cotton)
TBariknegaulpiqtuoo2t 5sammpLle(moeft5hmanLol(,ramnefaesrurpiinpgeftl)ask)
Water, 2 mL (ransfer pipette)
* Bring up to 10 mL (methano,l measuring flask)
Take aliquot sample of 2.5mL (transfer pipette)
Spih)
* B`mreiansguruipngtofl1a0skm)L (methanol water >* (8/2 V/V,
ory
`The percent recovery in each of the above operations are given below (sce Fig. 2).
Percent recovery (1=1) Peak I: 104%
Peak 2: 119%
Peak 3: (not measurable due to insufficient
Blank (n= 1)
LC-MS sensitivity) Peakl: notdetected
Peak2 mot detected
Peak: notdetected
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4.2 Quantitative Analysis
4.2.1 Quantitative Determination Conditions:
Apparatus:
Liquid chromatograph-mass spectrometer
Pump:
Autosampler:
PU-980 (JASCO)
AS-950 (JASCO)
Mass Spectrometer: Quattro II (Micromass)
Liquid Chromatography Conditions
Column:
L-column ODS (CERI); 15 cm x 2.1 mm LD.
Column temperature: 35C
`Eluting solvent: A (80%): methanol
B (20%): wate>r
Flow rate:
0.2 mL/min
Injection rate:
20 pL
Mass Spectrometer Conditions
Ionization method:
Electrospray (EST)
Detected ions:
Positive ions
Detection method:
Selected ion monitoring (SIM)
Selected ions:
Peak 1 m/z 571
Peak 2 m/z 671
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Ton source temperature:
P1e20akC3 m/z 771
Cone voltage:
50V
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Study No. 44102
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4.2.2 Preparation of Standard Solution:
sample`wSetraendparrdepsaorleudtiaosnsdefsocrrdiebteedrbmeilnoiwn.g the test substance concentration in the analytic
(1) Test water analysis:
dissolveOdnienhmuentdhraendolmitlolfiogrrammas o1,f0t0h0emsgam/pLlteesftursnuibsshteadncweassolpurteicoins.clTyhwiesisgohleudtioountwaansd
tPrheetnrdeiastsmoelnvteodpienrmaettiohnasnowler"e/wcaatrerrie>d o(u8t/o2nV/tVhe)troesuflotrinmgaso2l.u0t0iomngi/nLascoclourtidoann.ce with the
flow scheme shown below, `mg/L (for Peak 3) standard
stohleurteiboynsf.orming
0.0500
mg/L.
(forPeaks
1
and
2)
and
0.200
Flow Scheme
Column chromatography" (see page 6 for details)
2.00 mg/L test substance solution, 1 mL (transferpipette) Bmeraisnugruipngtofl1a0skm)L (methanol "water TM* (8/2 V/V),
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Takealiquotsampleof2.5mi (transfer pipette)
* Bmreiansguruipngtofl1a0skm)L(methanol */water> (812 V/V),
ary
(2) Test fish analysis: dissolveOdnien hmuentdhraendolmitlolfigorramams o1,f0t0h0emsga/mLptleesftursnuibsshteadncweassolpurteiconi.selTyhwiesisgohleudtioountwaansd tfohrenPedailkuste1dawnidth2maentdhaan0o.l20*0/wmagte/rL>s?ta(n8/d2arVd/sVo)luttoiofnorfmor0P.e0a5k003.mg/L standard solutions
i
(3) Preparationofcalibration curve:
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above, 0F.o0l2l5o0w,i0n.g0t5h0e0saanmde0p.r1o0c0edmugr/eLassttahnadtarudsesdoltuotpiroenspafroertPheeakssta1ndaanrdd 2s,olauntdio0n.s1i0n0,(2)
0u.n2d0e0r athnedq0u.a4nt0i0tmatgi/veLdsettaenrdmairndatsoilountcioonnsdfiotrioPnesagkiv3enweirnesepcrteipoanre4d..2.1Thaebsoveew,earnedaanalyzed
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calibration curve was prepared the chromatogram (see Fig. 3).
from
the
respective
concentrations
and
the
peak
areas
on
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Study No. 44102
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TTTSICRC of3Chbismaeulls2E0S0+3
- ia =~ 4
Co
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Peak
a5sve
2 pa Iik
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5
Bh 008
Sy R4ckelinet wakor Posvk1
543%
Poak2 6542
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iE a Safacame 5.
1.A30a0%
2
TB, 5 Tog
ag
af t
Blanktes oftotWeta?
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1.3004
| aes D3 w ot
PenPkepioksi2tion
{
No b>
| : z z E 60 1000 1200 "1403 Time
e327 A.Mahe
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Fig.1-1 Mass ragmantagramof LO-MS analysisfor acovery andblankfast
(analyssoftestwater)
Study No. 44102
10 Company Sanilized. Dassnot containTSCA CBI
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ii %
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------d ----a u 11T0i0ey8
Peak3
4
1E2S40
=14 8 |
45
0a=t
I
om
ast
8.
10.
& 12.00 14.00 Te
SiRof 3Char5l5 100T8I4C
jiw
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Peaks
1245
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. 548
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g IMT
4B Time
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Fghat BHR HE37
SRof annalTs8e5
i1
1.00e4.
iw :
i:
PeaPkepoissition
i
No, po
[had 7 Th0 Ed El TE Ee ET
2, 27 Nek
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Fig.1-2 M(asasnfaraygosmfetinetsstogwraatmerof LC-MS analyels for recovery and blank test
Study No. 44102
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LH io p
100.
Peak
SiRaraCh1a3m0nes T5i5e
30W0e8e4
T a430 i72e8
T [Rf| atIozCs8i1as72)Racava7r5y aecdhaom sia THT
1
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7+8|%0
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`Area
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No.
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Fig:2 MassragmentogramofLC-4S analysefo ecovar0y03a.ndb7i.a4n3ktPesLtMede
(anaolftyesstfiissh)
Study No. 44102
12 Comepanym, Sanitzec. Docs nolcontainTSCATE:
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Fig.3-1-1 Mass fragmentogrem of LC-MS analysis for calibration curve (Peak 1)
Study No. 44102
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Fig.3+1-2 Callbraioncurve of LC-MS analysis(Peak1).
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September5, 2003
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Study No.44102
14 Company Sanitized. Docs not contain TSCA CBI
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Fig:-2+1 Messfragmentogramof LC-MS analysis forcalibrationcurve (Fezk2)
Study No. 44102
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Fig.3-2-2 CalibrationcurveofLC-MSanalysis(Peak 2).
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Study No.44102
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Fig331. Mass fragmentogramof LC-MS analysisfercalbration curve (Peak 3)
Study No. 44102
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ozs
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Fig.3-3-2 Calibrationcurve ofLC-MSalysis(Peak3). Sepemter,2003 em. Mads
Translation: ~ Language Services
F. Metreaud
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December 3, 2003
Study No. 44102 --------------
18 Coftipany Sanitized. Does nol containTSCACBY