Document bBnkz1Lgenk9BVEB0QLGLqDG1
AR226_ 3306
488
_-- --~
AR 286_ 3306 DeDumpoincs11954
TRADE SECRET
Study Tide
F-24768: Influence on Growth and Growth Rateofthe Green Alga Selenastrumcapricornutum
TEST GUIDELINES: OECD Guideline for the TestingofChemicals
Section 2: Effects on Biotic Systems, Number 201(1984)
Commission Directive 92/69/EEC EEC Method C3 (1992)
AUTHOR:Terry Lee Sloman, B.S.
STUDY COMPLETED ON: October 31, 2003
PERFORMING LABORATORY:
E.L du Pont de Nemours and Company Haskell Laboratory for Health and Environmental Sciences Elkton Road, P.O. Box 50 Newark, Delaware 19714-0050
LABORATORY PROJECT ID: DuPont-11954
~
CompanySanitized. Dossnotcontain sca cr
TR
--
Ste DGorowweEthRRap tteoffthoeGr r-- een AAllgc saSelleenaascto pricr omutnmuin_ DDuupo_ ni11995544 --
`GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT `Thisstudywas conducted in compliance with U.S. EPA TSCA (40 CFR part 792) Good Laboratory Practice Standards, which are consistent with the OECD Principles of Good Laboratory Practice (as revised in 1997) published in ENV/MC/CHEM(98)17 and MAFF Japan `Good Laboratory Practice Standards (59 NohSan Number 3850). `The test substance was characterized by the sponsor prior to the initiationofthis study. Although the characterization was not performed under Good Laboratory Practice Standards, the characterization was done by an ISO 9000 certified laboratory, and the. accuracyofthe data is considered sufficient for the purposesofthis study.
Applicant/ Sponsor: E.I. du Pontde Nemours and Company Wilmington, Delaware 19898
USA.
--_
-- StudyDirector: _|
I
`Lee Sloman, B.S. Associate Scientist
31-0T 2003
Date
Applicant / SponsorT: DTibnRepeemaive Dame
--~ _--
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StGHr2oo4ww7t6h5:RRaatIteneofolffutehhneeceGGrornoeeeGnnrNAolngwaSaSneldleonanstirumrccaapprircoinuctuomrn --
QUALITY ASSURANCE STATEMENT Haskell Sample Number(s):
24768
Dates of Inspections:
Protocol: July 10, 2003
Conduct: July 23, 2003
Records, Reports: October 8, 9, 2003
Dates Findings Reported to:
Study Director: October 13, 2003
~
Management: October 13, 30, 2003
DDuubpoonnet1t1i9s5s4
Reported by:
Claer
Joseph C. Hamil
--_ H
Staff Quality Assurance Auditor
30dDate202-
Eompany Saniized. Dossnotcontain TSCA cI TTTTee 2
Ps GS2Grro4ow7et6h:ReattfeonfftethheaeGronrneesGnrcoArwltahseuaSndemesccpaprriciorncomuean___DDuur_pon_e119_564
CERTIFICATION
We, the undersigned,
from this study.
declare
that
this
report
provides
an
accurate
evaluationof data
obtained
AualytcalEvaluationby:
Bgay Sn SAC
`Bogdan Szostek, Ph.D. `SeniorResearch Chemist
30~0- 2c00T3
Date
Reportedby: le 3070. 2003 `StatisticalAnalysis
~~
John W. Green,Pb.D., PED.
Date
Principal Research Statistician
Ibys StudyDie rector: __1
Moor
"Terry Lo Sloman, BS.
AssociateScientist
31-0T-2003
De
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`TABLE OF CONTENTS
Page
GOOD LABORATORY PRACTICE COMPLIANCE STATEMENT cocoons
QUALITY ASSURANCE STATEMENT ..cccvnmssmmmssssssssmsseemsmesemsmd
Pi.
STUDY PERSONNEL ccvcvmmssmmsssnssssmsssssssmsssssssssssssssssssmssssond
MATERIALS AND METHODS mss
----------------------------sssssssssn 1)
A.
Bu
Test
TSE
SGuUiBdSeIlAiNnCeEs
..............
s sm --s ----------s ------------sssss 1100
F D. TES DESIGN wove T n 11
F. EXPETiMental DESIgN.......ovvvmeserosssssesssssssssssssesssssseosesenesssmnosn
nl
Gi
H.
TTeEsStECSonOdIiOtiNoSns.-..DcevfoiensiotvitvsessasnndneRsEeCseOsVeEsreyesTnsesssstsssssosoe.s.e.seesnsesesesese
NR
12
I. Selenastrum capricornutum Growth Measurement...
13
J. SHAUSHCA! AAIYSES coors essssssssssesesosssesesses esses, serra]
K. Analysis ofTest Solutions Definitive Test ..................oooom... csserernsessesssrnes 14
RESULTS AND DISCUSSION. r----------------------essssssnn 18 A. Environmental Conditions - Definitive and ReCOVErYTests.......ocovevrserssersmsrsessserns | 8 B. Test Substance Stability and Analytical Verification ............................ S---- C. Selenastrum capricornutum GrOWH........o.vvwveeeevooeseseesso rersessnsssnennen 19
RECORDS AND SAMPLE STORAGE r----------------------------n.2
--
APPENDICES wormsdd
-_-- BompanySaitzsd. Dontscontsain Toca cay
HGr2o4w7t6h8:RaIteonftfheloGnrueGerenowAnltghcaaSenedenastra capricorn --
LI OF S TABT LES
Dupont 11954
1: PHMEASUROFETESMTSOELUNTIOTNS.S.. 2: CHAMBER LIGHTINTENSITY,SHAKING SPEED, ANDTEMPERATURE
Page
23 RANGE... 23
3. 4
HMEEAALSTUHRYECDECLLOCOUNNTCDAETA SNOUFMTMH-A2R4RY76-8AIDNEFTTIENSITITSIVOOELUTTENISTOcNSSc...v.2827
5: AGRREOAWUTNHDREARTETHDEATGAROSWUTMHMACRUYRVEDEDFAITNAITSIUVEMMARY-TES. DEFINITIVE
TEST... a 29
7. T2HOUR (DAY 3) STATISTICAL ANALYSES SUMM-DEAFINRITIYVE TEST..
30
LI OF FS IGUT RES
1A
ATIVEANAL 5 ALBRATIONSTAND, VESioh isis
Page
PN
> 3
RREEPPRREESSEENNTTAATTIIVVEE C CHRH OMATROGROAMOMOFFTAHA CEALBIT LBARNAKOTCIOONG NTSRTR OALNSDA OALRUDSTMOOLNUTIoOvN...
mnidl =. 39
& 3
RTE2PHROEUSREHNTEAATLITVHEYCCHELRLOMCAOTUONGTRDAOMSEORFEASTPEOSNTSECOCNUCREVENTFROARTITOHNEDEFINITIVE
SOLUTION.
37 39
DTE2F-IHNOIUTRIVAERTEESAT.U.N.D..E.R THE GROWTH CURVEDOSERESPONSE CURVE FOR THE
T2571
+
72HOURGROWTHRATE
DOSERESPONSE
CURVE
FOR
THE
ssn d] DEFINITIVETEST v.43
LIST OF APPENDICES
A AADNUTMER DIUI MCONESTIN TUENTTS.
Page
1B. D-
7722HHOOUURRAHREEAALUTNHYDECRELTLHCEOGURNOTWSTTAHTICSUTRIVCESSTFAOTRISTTHIECDSEFFIONRITTIHVEETDEESFTIN.I.T.IVE1807... 72:HOUR GROWTH
47 3
RATESTATISTICSFORTHE DEFINITIVETEST...
.
56
--_
-
CompanySanitized. Donote contsain Tsca car
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STUDY INFORMATION
SwmonymsCodes]
+ H-24768 Haskell Number: 24768
CAS Regis Number [BNERY)
Duron 1954
Purity;
Known Impurites: --
Physical Characteristics; Tan 0 light brown paste Stability: aTnhaeltyetsicalsumbesatsaunrceemsetnabtislictyonwdauscteevdalduuartiendgbtahseedstond Sponsor: UWEliS.lnAdiungPtoonnt,dDeelNaewmaoruers19a8n9d8Company
Study Initiated/Completed: July 3, 2003 / (see report cover page) Experimental Initiated/Completed: July22, 2003/ July25, 2003
-
:
Bompany Sanitzes, Doss nor Contain TSCA cay
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STUDY PERSONNEL Study Director: Terry Lee Sloman, B.S.
Management: Robert A. Hoke, Ph.D.
Analytical Chemist: Bogdan Szostek, Ph.D.
Management: S. Mark Kennedy, Ph.D.
Statistical Analysis:
Management:
John W. Green, Ph.D., Ph.D.
Janice L. Connell, M.S., B.A,
C.LH.
Ecotoxicology Report Preparation: Wanda F. Dinbokowitz
Management: Nancy S. Selzer, M.S.
~
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CompanySanitized. Does
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SUMMARY
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The 72-hour results," based on nominal concentrations are as follows:
Healthy Cell Count
ECq NOEC
> 100mg H-24768/L 25 mg H-24768/L
Area Under the Growth Curve
NEOCEsC*
2>51m00gmHg24H7-62847/6L8/L
--~
Growth Rate
ECi
NOEC*
2>51m00gmHg-2H4-72648716L8/L
Nheoanlethoyfctehlle cdeofuinnitrievleattievset 1c0ontcheenbtlraantkiocnosnterxohli.biTtheedraef5or0e%,aorrgerceoavteerrygtreostwtwhasinnhoitbipteironfobramseedd, on
~
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As determinbeyd the Jonckhteoehretrceonndtoels.t
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Eompany Sanitzed. bossnotcontain sca cer
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INTRODUCTION tThheisgrseteundyawlagasSceolnednuacstterdumtocadpertiecromrinnuetutmh.eeffect of H-24768 on the growth and growth rate of
MATERIALS AND METHODS
A. Test Guidelines
The study design complies with the following test guidelines: + oOnrgBainoitsiactiSoynstfeomrs.EcoGnuoimdeilcinCeof-oOrpetrhaetTieosntianngdofDeCvheelmoipcmalesn.t (OECD) (1984). 201 Effects + CE.u3r,oAplegaanlEIcnohinboimtiiocnCToesmtm.unMiettiheosds(fEoErC)th(e1D9e92t)e.rmiDinraetcitoinvoef9T2o/xi6c9i/tEyE.C Annex V, Part
--_
B. Test Substance
The test substance, H-24768, was supplied by the sponsor as a tan to light brown paste.
C. TestSystem
Selenastrum study.
capricornutum,a
freshwater,
unicellular,
non-motile,
green
alga,
was
used
in
this
1 Original Culture Source UTnhieveorrsiigtiynoalfcTueltxuarse astouArucsetiwna-sAutshteiDne,paTretxmaesn7t8o7f13B-o7t6a4n0y.- Culture CollectionofAlgae - The
2. Culture Maintenance
ct`Toohntedhicetuisltotunudsrye,usmSeeedtlheionndatshfteorrsutSmuedclyae.pnraTishcteoruromnrugctaaunpmirsicmucslotrwunreuertseuwmceuwrleatusmraebidansnteadsitoneenrdipluuinzbedldeisr2h5te0hdemlsiLatmEeerrlaeetnnuvmirreeoy.nemrPernitoarl wflearseksasceopnttiacianlilnygtarapnpsrfoexriremdatteolfyre5s0hmmeLodifufimlteevreerdy(3= tfoil7tedra-sytse.rilTihzeed)flAaAskPs wnuetrreiefnitttmedewdiituhmfaonadm stoppers to permit gas exchange.
--~
e a Alsos known as s Pseudoke irchneriella subcapitata.
T TeT pCoompanySanitzed.Doss not contaimn Ttsncsi
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--~
D. TestDesign
The test below:
design
used
for
the
Selenastrum
capricornutum
definitive
and
recovery
tests
is
described
Organism
Nutrient Medium
Flask Volume (mL)
Solution Volume (mL)
Volume Ratio
v _Se--lenastrum capricornutum~~ AAP 7 250 50 sit
E. TestPreparation (Appendix B)
1. AAP Nutrient Medium Preparation for Selenastrum capricornutum
(sAdoAeliuPotinnoiunztserdia)enndwtamt1 eemrd,Liwouifmtthhmewiaxmsiicnprgroeanfputtaerrrieeednabtcyhstaaoddcddkiitnsioognl.u1 tmiToLhnoetfoveaoaplcpuhrmooexfiotmfthaehtee6lymmae8cd0ri0onumumtLrowiaefnsMt.birlsoltuoigc-hkQtto c1alcihterL wiittehr oadfdmietidoinualm)Miwlleir-eQ"usewdatteor.prAeppparroeptrhieatlaergperropvoorltuimonesso(f1tmhLeomfeedaicuhmsrteoqcukisroeldutfioornthfeor definitive and recovery tests,
rm
The medium pH was adjusted to 7.52 with 0.IN sodium hydroxide. For both the definitive and
AcreaAccPohvweniurttyhrtieaesnt0ts.,m2t2ehdjeiummuemcdeiflolurumluoswseaesiancfetithlateteerd-esfftiielnreiirtl.iivzTeedhaeunsdcionrngetcaCoiovnmeerrisyntwgeistthpsrtweh-eesrtereerssitulloitrzieenddgiffniilttlhreaert-irsoetnferrsiiylgsietzreeadtmosr
rinemtahiendianrgkuantuaspepdromxeidmiatuemlywa4sCpraonpderalcyclstiomraetd.ed to room temperature prior o use. Any
2. Test Solution Preparation
Prior to the appropriate
cinointcieanttiornaotfiotnhse
definitive to be usc
itnestt,headreafnignei-tifviendtiesntg,
test
was
conducted
to
determine
the
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--_
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Company Sanilized. Dossnotcontain rena mer
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~
3. Test Culture Preparation
pDuupeonnit 111s95s4
For each definitive control and test solution, 3 aliquots (50 mL each) were placed inseparate
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achieve the cells/mL at
desired nominal test initiation, an
concentration ofapproximately 10,000 Selenastrumcapricornutum approximate 0.340 mL (=340 pL) aliquotofalgal inoculum froam
cloongtarroli.thmically growing stock culture was aseptically transferred to cach flask, except the abiotic
F. Experimental Design
1 Definitive Test
`The organisms were exposed for 72 hours (3 days), without test medium renewal. Test flasks
were arranged on a lighted shaker table in positioned each working day. Each blank
a chamber in a non-systematic control, test concentration, and
design and were reabiotic control was
tested as 3 replicates. Cell counts were made
definitive test initiation (0-hour or day 0).
`approximately
24,
48,
and
72
hours
after
the
2. Recovery Test
~~
Since noneofthe definitive test
on healthy cell count relative to
concentration exhibited a 50% or greater
the blank control,a recovery test was not
growth inhibition
necessary.
based
G. Test Solutions
Solutions used in the definitive test were as follows: Blank Control ~~ AAP nutrient medium containing no H-24768
Treatment Abiotic Control
nominal 6.25, 12.5, 25, 50, and 100 `mg H-24768/liter nutrient medium nominal 100 mg H-24768/liter nutrient `medium; (no Selenastrum capricornutum)
H. `Test Conditions - Definitive and Recovery Tests
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Model SL445C7). The algae were incubated for 72 hours without test `medium renewal for the
definitive test.
environmental
pIalrlaummeitneartsioanrewadsesscurpipbleidedinbtyhecofooll-lwohwiitneg
fluorescent
table.
tubes.
The target
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~
Test
Initial Population
Illumination
Photo- Shaking Period Speed Temperature
(lumens/m'=1ux) (hours) _ (rpms)
2)
Definitive 10,000 cells/mL
0001010000 24 100 2442
Recovery variable; based on definitive test,
6000010000 24 100
2442
tferromminsaetleicotne-ddatyesctounts
_-- concentrations
IL Selenastrum capricornutum Growth Measurement
1 Definitive Test
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--_
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sttiumnetecda)chwedraye.reSctoatridsetdicailn tchalecsutluatdiyornescoorftdhs.e MEoCsst caonudnNtsOwEeCrewmearedebaasteadppornomxiemaantehleyaltthheyscaelmle
counts and nominal concenrations.
J. Statistical Analyses Tfohrecmacehanteshteaclotnhcyencterllatcioounntw,erareeeaxupnrdeesrsetdhereglraotiwvtehtcoutrhvee,blaanndkgcronotwrtolh.ratAellatcatlhceu7la2t-ihoonusrwienrteerval based on the nominal concentrations. Tfohremualrae:aunder the growth curves for each interval were calculated according tothe following
A=[05Xe0XtN[y05-%No(Olo + [00).5XX((1Vy2 +- 1N1o) x-(V2a9+)Ny - 2Np)]
--
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Dupont 11954
--~
thheefgorlolowwtihnrgaftor(m4u)laf:or each est concentration and for the controls were calculated according fo
yon o No)
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%l =_c.C5T x 100
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All testsof significance were at = 0.05.
.
K. AnalysisofTest Solutions - Definitive Test
1. Sample Collection
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~
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oling all 3 replicates for
-
ompany Sanitized. Doesnotcontain TCA cat
GHr2o4w7th6:RaItnfolfuetnhceeGorneeGnroAwltghaaSnldenastram capricorn
Dupont 11954
--
seaamcphloefst.heThceonptrHolwaansddteetsetrcmoinnceedntfroarteiaonchsoofltuhtieosnes
and day
taking an aliquot 3 samples.
from
eachofthe
`pooled
`The concentrationsof H-24768 in eachofthe test solutions were determined by`high performance liquid chromatography/mass spectrometry (HPLC/MS).
2 Sample Treatment
Ascnotnutadrlyoilqs,tuaotrtetsft(rdcaooynmc0e)tnhtaernabdtliasonunkbsm,cioatnnttredodla,fboi`rowtaoinrcaklciyosnnigtsr.sotloAcwkne,raaelnidsqautometsptlfcerodonmcoentnhtderaapytoi3oolnaesnddwreseprulebimcsiaattmtepseodlfetdhaet
the blank
analysis.
Back-up solutions also were provided for each sample.
for All samples were analyzed on
the dayofreceipt. The da3y exposure samples were centrifuged in the IEC clinicaclentrifuge
for 30 minutes inorder to remove algae. Aliquots ofall test solutions weretransferredto
asuatmopslaemapldedredvitaols5 fmorLanaaceltyosniistrbiyleL)Ca/tMtShe. tAilmleosafmspalmepsliwnegr.eAddidliuttieodna1l:1lyw,itthheafcoetlolnoiwtirnigles(amSpmlLesof
`1w:e6r.e25d)i,lu1t2e.d5 wmigt/hL5(0d/i5lu0t%edac1e:to12n.i5t)r,i2l5e/mAAgP/Lnu(tdriileunttemde1d:2i5u),m5`0bemfogr/e Lan(adliylsuitse:d61.:2550)m,g1/0L0(mdgil/uLted
(diluted 1:100).
~
~~
TTT
Company Santized.eDoes,S
-
GHr2o4w7t6h8:RaItenoffheloGnrueGerenoAwnltgchaaSendeles capricornutan
~
3. Instrumentation and Conditions
Dupont 11954
HPLC Instrument: MS Instrument:
Model 2795, Waters 2Q, Micromass
LC Parameters: Column: Mobile Phase:
Luna 3u C18 (2), 20 mm x 2mm
BA--2a5cetmonMitrailmemonium acetate in water
C25 mM ammonium acetate in acetonitrile
Gradient
Time(min) ~~ %A %B %C Flow (m/min)
0
00 60 03
5
0 0 100 03
65
0 0 100 03
651
0 100 0 03
85
0 100 0 03
851
0 0 6 03
12
0 0 6 03
Column Temperature: ~~ 30C
Injection Volume:
3spL
--
Column Switch:
1.25 and 7 min
MS Parameters:
Tonization mod:
Electrospray, positive ions
Capillary voltage:
35KV
Cone voltage:
30v
Source Temperature: 120C
Desolation Temperature:_ 500C
Data Acquisition Function:
IR of 690.5, 734.5, 778.6, 822.6,
7 oz: 0-7 min IR of 790.5, 834.5, 878.6, 922.7, C10o 22.7,l1066l .7 mi z; 0-n 7 minof890.6, 934.6, 978.7,
4 Quantitation
25436 and H:24768 chemically represen}
i
ing ciructure{CF~(CF),-CHy-CH-O-(CHy-CHz-O),-H, wher
The major compgnentsofthe mixture are derived for thrce
--
exc selected for cachoftel
Five of he gost intense ind For cachofth sidered
i]
five of the most intense]
epresente
The]
re
-
CompanySanitized. Boss not contain Tsca os
SSrDtoowEewRRatleeooff ebteeGGrrneee--enAMal--osnS--seers cauprmicocrnopriomanp_wDeupmo_enre1i1o_9s5s
~~
ionized in positive ion mode electrospray predominately as the ammonium fon adducts, when the
mobile phase contains significant
`based on the total signal obtained
concentration
for five select
of
ammonium
acetate.
The
guantitation was
Separate
s calibrato ion ce urves. were tmheadteestfsorubesatcahncgerowuapsofnot known.
The
totJalTchoepecnotnrtaetnitoonf
eoafcthhoef
the
test
substance (either H-25436 or H-24768) was assigned to each group oj-- were
`monitored.
A primary stock slutonofthe reference compound, ass as made in 10mLof acetonitrile. Calibration standards were pretedon cach analysis day by,
dilution with 50/50 %
1.03 mg/L. Duplicate
iancjeetcotniiotnrsioflec/aAlAiPbrnauttirioenntsomleutdiiounsmwaetrceonmcaednteraatnidonpsearkanhgeiingghtfsrwoemr0e.103
to
determined by electronic integration.
`The calibration curves were generated by regression analysisofthe peak height data obtained
from chromatographic analysisofthe
the calibration curve to determine the
calibration solutions. Data
concentrationsof H-24768.
for test solutions was fitted to
The limitofdetectio(n LOD)
and limitof quantitation (LOQ) were determined by calculating the average noise level in
chromatogramsofblank control samples and comparing them to the signalof a calibration
standard ofknown concentration. Two chromatograms were examined for `noise-related peaks
`near the retention timeofthe
equivalentofthe mean noise
analyte. The LOD was calculated
level. The LOQ was calculated as
as 3 times the concentration
10times the concentration
"~
equivalentofthe mean noise level.
--_--
Company Sanitzed. Doss not cantare vers nor
SeS GHr-l2o4w7te 6h8l :RaItl nefe olfuetnho ceeOGarnner enGnrAoAwltpgm haaSnldee nasiraam cpp ricorr nutumieomuten pwDrue_ pmoenlti_1o19s5e ~
RESULTS AND DISCUSSION
A (ETnavbilreson1m-e2n)tal Conditions - Definitive and Recovery Tests
For the definitive test, the pH the 0-hour and from 7.23 to 7.59
measurementsofthe test solutions at the 72-hourinterval.
ranged
from
7.49
to
7.68
at
*
For the definitive test, light intensity for the areas used 7440 lumens/m? (= lux). Themean light intensity was
in the chamber ranged 6800 lumens/m?.
from
6250
to
* For the definitive test, the shaking speed was 98 revolutions per min (rpm).
* rFaonrgtehde fdrefoimni2t4i.v7e ttoes2,4.th8eCtefmopretrhaetudrueraftrioonmothfetdhiegsittuadly.readout on the temperature recorder
B. `Test Substance Stability and Analytical Verification (Table 3, Figures 1-4)
1. Chromatographic Results
leof Te
luted as a well-resolved peak with a retention timeof approximately
2.0 minutes.
`approximately 3.6 minutes.
luted as a well-resolved peak with a retention time of
luted as a well-resolved peak with a retention
imeof approximately 5.0 minutes. Figure 1 shows representative calibration curves obtained for
typical standards are shown in solution and test concentration
Figure 2. Figures 3 and solution,respectively.
4
show
chromatograms
of
a
blank
control
o`fThHeL-2O5D43a6n,dreLspOecQtifvoely. The LOD and
ere
Co --
determined
to
be
0.905
mg/L and 0.016 mg/L
determined to be
0.002 mg/L
o--
and
0.007 mg/L
determined
of
to
H-25436,
be 0.019
mrges/pLecatnivdel0y..06T2hmegL/LOoDfaHn-d254L3O6Q,
oP
respectively.
2. Test Solution Results
`The mean,
(= nominal
measured concentration
100 mg H-24768/L est
|--
concentration,
=
abioti
the da0y working stock
control) was 110 mg/L. This
|<Rmaeas L
ecoveryofth active ingredient. The in the day 0 working stock (=nominal
mean, measured concentrationofih) 100 mg H-24768/L test concentration,
= abiotic control)
mean, measured
was 95 mg/L.
concentration o
This represents
--
95%
recovtehreyodfatyh0ewaocrtkiivenginsgtroedciken(t=,
The.
nominal
"roLommpanySanitized. Does not conta vra ny
G1r1o2w0t7h68RtnefoutehceeCorneGmroAwlthsaSlam pric
Dureniioss
--~
r1e0c0ovmegryoH-f2t4h7e6a8t/iLvteesitncgorendcieennttr,ation, = abiotic control) was 99 mg/L. This represents 99%
concenraionso SRR sor 0 pomme 635 125 35 mS "The mean, measured concentrationsofESSER: the day 0 nominal 6.25, 12.5, 25,
and 50 mg/L test concentration solutions were 6.3, 12, 26, and 52 mg/L, respectively, This
represents 101, 96, 104, and
measured concentrations o
104%
recoveryofthe
actthievedianyg0rendoimenitn,arles6p.e2c5t,iv1e2l.y5.,
2T5,heanmdean,
50 mg/L test concentration solutions were3.4, 11, 23, and 47 mg/L, respectively. This represents
86, 88, 92, and 94% recovery of the active ingredient, respectively. The mean, measured
concentration solutions were 5.3, 11, 24, and 49 mg/L, respectively. This represents 85, 88, 96,
i penn, he bint ent oo omens scacoe) mcaonendcne9n8tm%reaatrsieuocnroevsedorlcyutooifnontcshnewieacrotesivpoereiR pnagrreeddiaenR tt,hreedsepsR eicrtievdelnya: .miTnhaelsceoondcaetnmatriantdi4ioc3nast5.e1d 2Ath5fe2rs5to5cdk0aya.sn,dmhdteest
100 mg/L test concentrations, and abiotic control solutionswere4.3, 8.9, 18, 35, and 73 mg/L and 75 mg/L, respectively. This represents 69, 71, 72, 70, 73, and 75% recoveryofthe active
ninbotthhe dnaoymi0naalnd6.d25a,3y1.2.5A,ft2e5r, 350d,ayasn,dth1e00memagn/,L.mteeastsucronecdenctornacteinotnrsa,tiaonnds of
rabeiporteiscenctosnt5r1o,l
5s7o,lu6t4i,o6n4s,w6e8r,ea3n.d2,679.1%,
r1e6c,o3v2e,ryaonfdt6h8e
magct/iLveanindgr6e9dimegn/t,L,rersepsepcetcitvievleyl.y.
This
The blank
~
control solutions
and day 3. After
c3odnatyasi,netdhenmoedaent,ecmteaabsleurceodncceonntcreanttiroantsioofns
of
n both day 0
in the
nominal 6.25, 12.5,
72.495,47r.e1c,o1v6e,r3yo2,fatnhed
25,
7ac2t
im5vg0e,/Liannagdnred1d0i7e04ntmm,ggr//eLLsp,etcertsietvsecploeycn,tcievnetTlryha.etiboTlnhsai,nskarcneodpnrtaerbsoielonttsisocl4cu6ot,nito5rn7os,lc6s4oo,nlt6ua4tii,noe7nd2s,nwaoenrde
detectable conenrutions ofREE bo do 0 and doy 3
iS hiro RTehceosvtearbyiloitfyS of H24768 iE s difficult io R assess since it is composed{ crcl onJ 6 cR s n
measured concentration were 68, 73
%ofday 0 measured concentrations. These results
suggest marginal stability ot es
CO d concentR rations a. s the bencihmcartk
cx the 3 day study period using 70% of dau
cfohristnabsilictoyoinndianyg3S.)eRlecoovnerices oovoen
0
ranged from 63.74%of day 0 measured valyes with the exceptionofthe day 3 recoveries for the
RoNncenration.
C Selenastrum capricornutum Growth
(Tables 4-8, Figures 5-7, Appendices C-E)
IL Definitve Test
The organisms were exposed for 72 hours without test medium renewal, The effets were
--_
expressed in termsofpercent inhibition in growth based on healthy cell count, area under the
-19 &Lompany Does Sanitized, notcota TSA cy
GHm r2o4w7t6he 8:RaItnefe loufetnhceeCGoreneneGnrNoAwltgghaSaSneedlleneasaisrummccoapprriciocrnoutmuumun_____
DDuupoPnotn1t119159544.
~
gbarsoewdthoncunrovme,inaanld
tgerstowctonhcreantterarteiloatnisv.e
tToheth7e2b-lhaonukrcroensturlotls
for are
the 72-hour as follows:
interval
ofthe
test
Healthy Cell Count
ECy NOEC
> 100 mg H-24768/L 25 mg H-24768/L
Area Under the Growth Curve
ECw NOEC
> 100 mg H-24768/L 25 mg H24768/L.
Growth Rate ECs NOEC
> 100 mg H-24768/L 25 mg H24768/L
Tdohseer-eddeupcetnidoennstirnehsepaolntsheywcieltlhcionucnrte,asairnegacuonndceernttrhaetgiornoswotfhtchuervtee,stasnudbsgtraonwcteh. rate indicate a 2. Recovery Test
--
`heNaolnteohfytcehlel cdoeufinntitrievleattievsettcoontcheenbtlraantikocnosnterxolh.ibiTtheedraef5or0e%, oarrgerceoavteerrygtreostwtwhasinnhoitbipteirofnobramseedd.on
CONCLUSIONS
STienhclereenraaesdsiutncrgtuicmoonncscaepinrntirhcaeotarilnotunhstyoucfmetlathce7o2utnehtso,usarursbesa(t3aundncadeye,sr)Ht-ih2ne4dg7ic6ra8ot.wetahHdocouwsreevv-ee,drea,pnetdnhdegerEnoCtwytryhesifspoogrnrseeatfeorrthan
100 mg/L based on nominal test concentrations
-
RECORDS AND SAMPLE STORAGE bLeabroertaationreyd-sbpyectihfeifcaocrisliittye-wshpeerceiftihcerwaowrdkatwza,ssudcohnea.s personnel files and equipment records will NMSoepuwenactriakmi,ennDse(Rleiacfwoaarprdpesl,iMctaahbnelaeDg)u,ePmroeannwttd,CatrWzoi,lpamPnirdnogtttehocetnfi,ionnDaelAlrracewhpaiorrvete.sw,ilCNlheabwreaacrrtkeet,raiDizneaeltdaiwaoatnrHdeaa,stkaoera(lplterLIcarebonontraptuorriyty,, JcaocmpkossointLiaobno,raatnodrikens,owDneeipmwpautreirti,esN)ewwilJlebrseeys.tored at Regional Analytical Services (RAS),
--~
TTT
CompanySaitzed.Doesnot conten 75a op1
nism
--~ SoSGHu-r2t4oe7h6w8R:RlaatItenefooolfufettnhhnceeeGGroresneeeGnnrtoAAwllgtgaharaSnldeunasmtrunccaporicpomutrum icomuton__DuDPu_poonnt_t111_1995544
REFERENCES
1. AMislslaeyr,BWot.tEl.e;TGerset;enUe.,S.1.EC.n;viSrhoinrmoeynatmaal, PTr.otTehcteiSoenlAegneansctyr;umUc.Sa.prGiocvoerrnuntmuemntPrPirnitnztiAnlggaOlffice: `Washington, DC, 1978; EPA-600/9-78.013.
2. AComnedruicctainngSoSctiaettiyc f9o6r-hTeTsotxiincgitayndTeMsattserwiiatlhsM(iAcrSoTaMl)g.ae"(1i9n90A)S.T"MStAannnduaradlGBuoiodkeoffoSrtandards, E1218:90. Vol. 11.04, Philadelphia, PA.
3.
Snedecor, G.W. and Cochran,
`University Press, Ams.
W.G.
(1967).
Statistical Methods, 6" edition.
The lowa State
4. pTraomcheadnuer,esA.foCr.,idDeunntnieftyti,ngCm.aW.x,iGmruemensaJf.eWd.o,sea.ndJWAemtehre.rinStgattoisnt,, AJ.sDs.oc(.209061,).835M-u8l4t3i,ple test
5. OU.PSP.TESnv8i5r0o.n1m0e0n0t.alSpPercoitaelctCioonnsAigdeernactyi.on(s19f9o6r)C.onEdcuolcotgiincgalAqEufafetcitcsLTaebsotraGutiodreylSitnuedsi.es, Public Draft. EPA 712-C-96-113,
--
~~
CLoompanmy pany,sq,1220. Dossnotcontain car
-
CGH-rr2oo4wn7t6h8R:RoavtIeeoonffthfheelGoGrnrueeGeenrenoAAwngltgshcaaSSneeedll.eonraassttroummpcraipcrorimcuotrumn _______ --~
DDuubPoonn-t1111995544
--_
TABLES
--~ Company Sanitized. Doss notcontain TSCA CBI
eres
StG2r4oo7uw6th8h:RReatItnefolofuftethnheceeGGorrneeeGennrAoAwgltgshaSaSneldlennaassiurmucmapcraiprciocromruntuotumm ____ --
DuDbuoPnotn1t119159544
TABLE 1
PH MEASUREMENTS OF TEST SOLUTIONS
Nominal H-24768 Concentration Blank Control 625 mg/L 125mg 25mg 50 mg/L 100 mg/L Abiotic Control (100 mg/L)
--_--_--Exposure Initiated: 22-Jul-2003 0-Hour (Day 0 749 751 7.68 7.60 7.54 7.59
7.59
Exposure Ended: 25-Jul-2003
72-Hour (Day 3) 7.49 7.56 759 7.59 7.58 752
723
TABLE 2
CHAMBER LIGHT INTENSITY, SHAKING SPEED, AND TEMPERATURE RANGE
`Mean Light Intensity
Test
at Test Initiation ~~ Oscillations
(umens/m? = ux)
(rpms)
Temperature cq
Definitive
6800
9%
2710248
--~
Company Sanitized. Does not contain TSCA CBI
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~~
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2;
--~
5 i
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Z3G
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+
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CompanySaniized. Doos not contain TSCA CB
~
TABLE 4
HEALTHY CELL COUNT aDATA SUMMARY - ie DEFINITIVE TEST EE E E = e = I e 2 = & r Ne == E = = e se e =s = - -- E = e = _ =F e r = = -- F e o -- =FF == E = s=r=o=e e= = 1 = = r S = = C = == = ==t r =-- == = c == --= = = = e = F r =5=
--
Se-- Company Sanitized. Doss notcontain TSCA Cs;
~ E --_ T su
TABLE 5
AREA UNDER THE GROWTH CURVE DATA SUMMARY - DEFINITIVE TEST
wm Ee doE m Nominal
Der: BI
rs Under oeGo Cor
00
= == = r = =
e = = = e---- --r -- r---- --
SE e = = = -- == --= --= oben]
Sioa
amoed
rm
= = = ee r e i ese
sa
ma
55
= = -- EEE r r =
= = CT] = = r r = =
~ I.oT Company Sanitized. Donote contsain TSCA oa
pe
ie
TABLE 6
--GROWTH RATE i ATe DATA SUMMARY - DEFINITIVE TEST
Concentration
He
eS
"-
73 Hour
= S= [ ==c== --= =r == = ] --= ray
= e Eo==T=T]F=1--== r == = I=
EE-e=T = =R-- a &-- wr =
= = =- E===E=ET= &en = &wom =&
L= =E e==T= E E -- r E= E SE = o
= = Se E=E== eE=TT= =--==FF=--=
~
Company Sanitized. Does not contain TSCA CBI
1Gr2o4w7t6h8:RaIt noftfhelaGnrueGeernoAnwlgcaaSenedlenasrum capricornutum --
DuPont 11954
TABL7E
72-HOUR (DAY 3) STATISTICAL ANALYSES SUMMARY - DEFINITIVE TEST
Nominal
TReesstpCoonnsceen(tmrga/tLi)on
ConBce2n4t7r6at8ion _ ReMsepaonnse StEarnrdoarrd N| Leva Value
CHoeulntbyCal 6B2l5amkCgowol 22233730305030 112444293512 33| ECo >100
12255mamlall.l
217606333333 111908905158 33| NOEC 25
S1o0m0mag.
B196n667 3284042 33
Area Underthe BlankConrol ~~ 38880000 2.163331 3 |
GrowthCurve 162.255mmgglL 92424800000000 44116062344817 33| EC >100
2S5ommaelrl
229192200000000 21316954360575 33| NOEC 25
100mg
2280000 1028397 3
~
GrowthRate 6B2l5anmkCgontrol
000077819 00.00000097 33| ECo >100
2152m5gmg
0000777170 o00m0i0t9 33| NoEC 25
So10m0am.g.
00006784s 0000000013 33
~~ -
Company"s,Sanitized. Doesnotcontainrca gy
HGr2o4w7t6h8RaInoftfheloGneuGerenowAntlhscaSneldenasirum capricorn
DuPont 11954
--_
FIGURES
-
Compa#ny1s,Saniized. Does not contain Ts0a may
GH.r24o7w68:RatInefolfutehneceGorneeGnroAwltghasSnedlenastra capricorn --
Dupont 11954
FIGURE 1
REPRESENTATIVE ANALYTICAL CALIBRATION STANDARD CURVES FOR H-25436
A8 c
Compound namSe]ERIIRT Goeffcieonft Determination: R"2 = 0.999185 Calloraton curve: -3.317766+006 * x2 + 1770764007 * x + 154110
1491554
|
||| 25 A
|
~~
E
2
: | 0.00 0.20 040 0.60 080 100
H-25436 Concentration, mg/L
~
`Company Sanlized. Doss not contain TSCACBI
-
Gc GHrr2oo4ww7t6h8R:o RaattIenefoolffutehp nheceeGGrorneeeeGnnrr AoAlwlgtgahsaSSneei dleenraassrruuc mm caprco ornutanmum___ DDuuppoonne_ 1111995544
FIGURE 1 (Continued) REPRESENTATIVE ANALYTICAL CALIBRATION STANDARD CURVES
FOR H.25436 Al B c
--
2]
Goeffcent of Datermination: RY2 = 0.998545
Calloraton curve: 1 277594008 * x'2+ 1.187916+007 * x + 699788
11008090
| | g
|
|
--_
E5
|
&
| 0.00 020 040 0.60 080 1.00 |
H-25436 Concentration, mg/L
|
------------S--------------------
Company Santzod. Does not conan T5ca ent
=
--
GG1rr.o2ow4wt7t6hh8R:RataIetnfoolffuettnhhceeGGorrneeGennrAoAlwlgtgaha aSSnledelneansarsurmmccaaprpcrorimuctaonrn --_
DDuuPpoonne1t11919854.
FIGURE 1 (Continued) REPRESENTATIVE ANALYTIFCOARLHC.A25L4I3B6RATION STANDARD CURVES
A B c
Gompound name} osfiien of Determination: R2 = 0.998745 Calibration curve: 223005 x2 + 45162764006 * x + 16260.8
wong
:5
~~
2
&i
| 000 | 020 040 060 0580 1.00 |
|
H-25436 Concentration, mg/L.
~ `CompanySaniized. Doss not contain TSCA cai
--e-- rie ----ee ER stm----
SrHG2ro4ov7w6th8RR:aattneefoooffnttchheeeGGroreneeGenrnoAAwlgtghsaSaSneedlnlaesrnumaeqcparpicroircmoatronn --
bDuuroonn 1111995544
FIGUR2E
REPRESENTATIVE CHROMATOGRAM OF A `CALIBRATION STANDARD SOLUTION
[o.7220381 _--
||
371091351841.3981
FISRols chamesTgeSs || et ad
N
--
To 26 360 abo 550 55 min
C2O5I413.c3on.c6en . co0.1o03mnL. srry 2.0 mins. Cation snr sion vs
rEg wo
-
FESR Sa12m91a00E0nS0eT8
aTnsanvaidsse
|
|
ol TH 7m shg a sh FaEwE mn CoTSm Cone ai-o-natorfet0.e1n0t3imongtime ofspproximatey 3.6 minutes. Calibration standard solution contains
ee `CompanySanitized: Doss nR ot conI tainTA 5c4 car
GGHrrAooAwTut6h8RR:aaftelooffetheeeGrraeneeGnnrAoAwlttash SsSneldlrensrmccappirioormam ~
pDuwroowni1i1o95s0e
FIGURE 2 (Continued)
Cz -
REPRESENTATIVE CHROMATOGRAM OF A
CALIBRATION STANDARD SOLUTION
Eo " mperoz
FREcomTetnse
)
|
> The Ei
I"
~ C2o 5t6sm enc3entm t6ion o5f.r0.e10c5 moLn.ieofsprovimatly5.0mits. Ctiteon sand soon coming
--~ -_----
CompanySantized. Doss notcontain Tsca a)
G1r2o4w7h6:RaIenfolfuehneceGarneGenroAwltah aSnldeasrum capricorn PS
Dupont 11954
FIGUR3 E
REPRESENTATIVE CHROMATOGRAM OF THE BLANK CONTROL SOLUTON
rm 0
EE20E 00Tso
*
Me
| I HR R a R
--~
mmm] cst eco inappl 20 mites,
er ww
EEETo aston
LHL I IR RoR
[mm]... elute at a retin imeofpproximately 3.6 minutes.
--~
a ompanySanitzed. Dossnotcontain cai
HG24r76R:aotIenofwlfutehnteceGroheneGnroAwltghaaenednasrum capricormutin --_
Dupont 11554
FIGURE 3 (Continued)
REPRESENTATIVE CHROMATOGRAM OF THE BLANK CONTROL SOLUTON
==
TR FE
0
wae
|
|
1.00
200
300
4.00
5.00
8.00
m
--_ [omen]... elute at a retention timeofapproximately 5.0 minutes,
Epmpany Saniized. Doss not contain Ta car
EE --
SrGHr2oo4w7lut6h8tR:naetnefouofaefttnhhceeeGsGorrneeeGtenroAwltgguphaSnedmenasctrumacaprpicorrmutiicornaer_DuD_buPoow_n1t11_919s54e --~
FIGUR4E REPRESENTATIVE CHROMATOGRAM OF A TEST CONCENTRATION SOLUTION
raz 0.
--- 2
To2s2i0rr7a3zs
FIR of shasaszmene0S0s5o87
|
|
o TR Tan T-h ah ah s"k min
C ta nominalconcentrationofY 5625migons(dioefds11s2.5obeforealmyi)s.s. Tos shinsele20c76
[orzo 10
Io a
FZSR ors amas 357
T0Ssoarerezszn
sos2e1005
*
Th 2-h we = ab she em min
T3AnomiNnal cIoncentAratio:n ofr62e3 nmgi/Ln. t(dsieofdsp1r2o.vbseafoeryan3.6amlyi)s, To soto se sma H2465
--_
-
Gompany Santizod. Doss not contain sca cer
SSG2or6ow7t6hlRh:usotorfhbneGrosrn eGne nroANr wlisp sSndmlsocaprcom icorstnpDu_upreomna1_1i9o34 ~
FIGURE 4 (Continued)
REPRESENTATIVE CHROMATOGRAM OF A TEST CONCENTRATION SOLUTION
a SREEST C
Te |
sTuoios 35
5.01
191024006
'
|
L%T e Te o m o e a |min, ~ o S008 monm Scnesionecanfc o3n3mioEeo(fhpevodsi: 1mic5ty55mitaes,.Testshin splcon
ps --_--
TTPenTees `Company Sanitized. Does notcontain TSCA CB
C142r0765s: heofreGarn GnrS o ntscrm _
Dupe 3se
FIGURE 5 72-HOUR HEALTHY CELL COUNT DOSE-RESPONSE CURVE FOR THE
DEFINITIVE TEST
TAT
AN~
|
WEI
|
0 Ee
2
--_
|
|
concen BHT G00 50 10
ES
0d
0 pot ton ;
..
I
|
= --_-- Company Sanitzad.e Doss noe t containTTescca cay
2o4n768RtaosCoonGosvatn rict
Duro ion
FIGURE 6
72-HOUR AREA UNDER THE GROWTH CURVE DOSE-RESPONSE CURVE FOR "THE DEFINITIVE TEST
hant rdhi e rote rnd
eonconrbit480 T 1TH 50 0
Nt
|
\
SUE 13 a > aN
|
=
Po
-- Ea
Er
WT |A
TT
i ay 4 0pt evn,
tN
:
ee]
--_
CompanySantized. Does not contain TSCA C51
- areStTescro Senses bron
FIGUR7 E 72-HOUR GROWTH RATE DOSE-RESPONSE CURVE FOR THE DEFINITIVE TEST
ah
ETIIN NG
|
S m TNS> ee
|
wd
~~
|
i
~~
~~
an i FA
* ow ow own om ww me
ef mt
rs
a
:
|
end2l
I TT |
ani
~~ TO
|
iE
NY
~
CompanySanitized.Does not contatn vena an
HG-r2o4w7t6h8:RaItnefolfuetnhceeGorneGernoAwltghaaSnedlenasrun capricornutum --_
Dupont 1954
--
APPENDICES
--_ _--
ar Company Sanitiezedd.DDoocsesmnoott,ecoonnttaaiinnTtSsCcAacycpr
-- SoSGH-ur2ote4w7th6h8Rl:RaavIteeneofolfuftnethnheceeGaGorrneeseeGnnrAoAtlwlgtghaauSalnedmnasctrumocappricrorniutumcormuten___D_uDPuoP_noen_1t1191_59544
~~
APPENDIX A
AAP Nutrient Medium Constituents
--~ _--
CompanySanitized. Doss not contain TSCA cp: ETH
1.24768: Influence on Growth and --_ SrSovtehRtleofaheGs reent AgsrumapricomutonD_upo_ m119_34
--A--A-- PNUTRIAAEP NNUTTRIMENET MDEDIIUUMMCOCNSOTINTUSENTTSIT~U~ENTS
ComponenSttock SolutCioonncsentration (1) CompoFniennaltPreCpoanrceendtMreatdiiounm(mg/L)
Macronutrient Stock #1
NaNO;
25.500
Na
11.001
N
4.200
Macronutrient Stock #2
NaHCO;
15.000
|
2.143
K
0.469
Macronutrient Stock #3
K;HPO,
1.044
P
0.186
Mg
2.904"
Macronutrient Stock #4
MgSO04#TH,0
14.700
--
Macronutrient Stock #5
MgCly*6H;0
12.164
Cas L1.o202
CMaaCclrozn2utHr:ie0nt Stock #6 4410
ComponenSttock SoluCtoinocnesntration (mg/L) CompoFniennaltP_reCpoanrceedntMreatdiiounm(ig/L)
Micronutrient Stock
H;BO;
185.520
B
32.460
MnClp*4 H:0
415.610
Mn
115.374
ZnCl,
3.2m
Zn
1.570
CoClz*6 H,0
1.428
Co
0.354
CuCl+2H,0
0.012
Cu
0.004
Na;MoOs+2 H:0
7.260
Mo
2.878
FeCly*6 HO
160.000
Fe
33.051
Na;EDTA<2 H,0
300.000
--------sm-- s ----------eee--
~
ba IAnscrleupdoerstmeadginneRseifuemrernocem|.both magnesium sulfite and magnesium chloride.
_--
Cosmwibe`Sasnitommcsminrseacs
GSGHrr-oo2w4wet7t6hh8RR:laaIttnofonlffutenheecaeGGrornoeseeGnnrAAioglgwaaraSnldeoastmra ccaporicpornrcornon___DDu_ubPoon_en1t119_15954. --
APPENDIX B 72-Hour Healthy Cell Count Statistics for the Definitive Test
--_ _
=TeSeohmpyanSyaSanniitiezedd.DDoosessnnoottoooonmtaaiinmTrsses re
5m26v7t68RtnaofbeeGcroeGnoAgtsoerdas crc ~
Drees
--2H72O-HUORURHHEEAALLTTHHYYCCEELLLLCCOOUUNNTTSSTTAATTIISSTTIICCSSFFOORRTTHHEEDDEEFFIINNIITTIIVVEETTEESSTT _
sSp0o0GnrostCmmeraeimSazhsrsameeermneOn,naic1as9m0rsf4or3oennrHSsckioenvmoiemseeSHenSeavors
Gone dEosoewalCONE ve wom som sma
pPDoo0ms0sTaE3l so
mBHmEGummRmss mmr
mBhneOanoRnw nnn
jammpsenkenis nan
nddseaenwsmss dome
ERNE 1 SH
sSpRoRseRsCamrRNaeeImaimLserEeerESoo,eSn iaI0sN3nrSeRsOsneRsMARUvLiLLseLRe oDrreuSneSiooovnns
ES mesos
sa ose
mm seems nous swe owes oss osesss
~
soSpoALsEVeVsEECmRaSrSeezCTsaFoenoaTnoonAnF,oNRaLCvIdEaeSnrtTsFRvOSiLsoGe,BrrraunoSedaasvnevis
sist or me sow sve ws ene oes
sors
aSnheandaiaicyaeiwSaSsuSEfeoIuTndaErttoEabneaHniormnaelypesroerenievsa.ces
orSOSc8RaRASsSCm0aIreeSzaesrOeaeFrsoonS,nhTra1hnm9ise0srsrsTezRnmsei+30ns,BneohbporiSesioecns
SE kil nl
Tae a2iess
Count 70K FULL DATA Ting cLiImNaE,NSroPnsRsOrUsTsoonntshonasnce Sations 33
ese wBo Hom mane se
--
Poms sn me con
-_--
Company Sanitized, Does not contain Tse mms
GH-r2o47w68R:atIenoflfuetnhceeGorneGernoAwltgshaSenldenastrum capricorn
Dupont. 11954
--~
LSUEANS
REPAONRATLSYCTSRAIETSAITSEOTDFICAOSNLELAE1N7NA:A1LS5YTSRIOTSNHUR-1S1OFsUAtLYLPrD3iA3sT0A0103 - DOSE WEANS ADJUSTED FOR THE EFFECTS OF SELENASTRUM
Count FROM FULL DATA Using Weight-NO WEIGHT and Variance Aurio3np
obs eves
Luan
= oor
210sas i FR x
226703030301.3030 2703333133
11222119964.040 1miselid
1172 1
5 dsooo
I1s6s6e3s3s3e3s373 19333333
112m2i1s9a6l4i4d 1zselid
1135 13
Count FROMESFTUILMLATDEADTUASDIOUNSsGEinAEgLFZFWHEeACi=Tg.S0h5t&eNDFOOURNWNECEIOTMGTPHATFROaIRnSOdDNevScarr7es0aasuCioOnngNTRmAOoLLrTtEoRsN0A.T3I3VE
Estimate
SIGNF
Dlu-nsniedtesd povalue
TesHtaacnrow
w
DD0oSsEe QTURAEDN DOSE 2-1
++
o0.l0o00e0s1 03120
2233330:.33
:. 3
~ DDDDOOOOSSSSEEEE 5463--.1111 ss+o olco0lol2oo5oo1oss5si6ss i1l2s73se03e33sn3s33ei33l03en33r3 33>
Count FREOSMTIFMUALTLEDDUASDTIOANSGEUsAEiLFnPFgHEAC=WT.eS0i5g&htFD=OUNRNONCEWOTEMTIPGAFHROTIRSaODnNedScrVeTaoarsiiCaOnnNgcTeRAOLLmTaEtRiMoAsT0.IV3E3
HMCaahxxeiicnmkuunnfoNNruummNbbueenrrbeoorffoRRfeeppRsseppsienriAnGnrycoouuT.nrteisstme3nt Group or Control
Exact Permutation Methods Recomended
AATnNeaAslLtyYsSfiIosSr UTo RESNDINp G AILNm PRALE. IOT.HE0R5DIRECTION in Response=count
FULL DATA
oKeTy Z_0T
I1ss
SJToAnNcDrAiREDEIRZsEDsmJaOzNzCsKTHiEcSRE
STATISTIC
PXX-PPVLRA__LJJUTTESIISSARP2E--VUFAAOLLRUUEETIFFSOO-RRCOUDRPORUWEACARTDREDDTRTTEERNSEDTNDWITH CONTINUITY SIGNI? RESULTS ARE FOR D ALTERNATIVE HYPOTHESIS
CORRECTION
FACTOR
--~ -
CompanySanitzed. Does not, contain TSCA CBr
GHr-o24w7t6h8R:aItneoflfutehnece GorneeGnroAwltgha aSnedlenastrumcapricornutun
DuPont 11954
--_
Jonckheere Trend Test on Dose 0 + Lowest 5 Doses thru 100 G/L
om. 31 3.466Z3I6T8 0.0X0P0L13O6T XeRJT S=iGNIF pose5
Jonckheere Trend Test on Dose 0 + Lowest 4 Doses thru 50 MG/L ar,295 -2.281Z1I7T2 0.0K11P4L3O8T8 XPRUTLoSeINTF Dose5
Jonckheere Trend Test on Dose 0 + Lowest 3 Doses thru 25 MG/L -- 28 -1.278229078 0.1X6P6L34O2T xeRT siGNIF pose4s GrJrooonrucpkhemsearnestseshtoulrdesubletsexaamrieneidncltuodcehdecikn tfohre lsaucmkaoroby-ftiarble 0 a linear trend before trend test results ave accepted.
--
--_ - Company Sanltized.Doesnotcontain sca csi
mS GHr-o2e w47tl6h8R:naItenefole futes nhceetGorenu eGnenrNoAwm letghaaSnc edenasotrup capici ornuc tum omutan__ DDuupPo_ own1t111_ 995544.
--
EC50 for Cell Count (Note: Formulai-cell Count/100000)
ROEBSSPEORNVSEED: &FOPRRMEUDLIACLTEDfroPmERD CENTACHAT NSGeE A FROMSCOE NTROT L MEAN
obs mcaolx x maw SED orem eReveorcENG
12 30
0.2050 wl
33 3
223473.03030 20033
234353..5603 220.312
=0 1
5o -10
6&5
s50l00o0 wooo
33 3
ssleaayn sls
lmleeriiisaslo enoaasy
"3129 "a
CBoCv50ariuasnicneg MMeatthroidxeMaAnRdQUSAtRaDnTd,ardBruBcrevor& VofersPtaereagsetWeeirshBteesidnaptreosbit
NMMEL logecx gama
ss
vglooagmeacx
200.1.000858507319094
00..4005909149 7.2425
27.902805 3531312
o0l.e2d9s6s5 18.8513
Hodel Parameter Estimates, 3050 and 95% Confidence Bounds
logecx gamma
yo
a
w
ses0
--
4.75450 1.76051 205.564 6.9268 207.625 136.108
MSoRdEeGl EvaluSaStiRon MeasuSrsetsorc ssromu square
391 Te.6661 237.796 3669.67 0.66919
nGroatpeh descriptions formulal: ECS0 3VP
FiInTcluBdYeMAtRhQeUAfRoDlTl,owing:
-- -
Company Sanitized. Does not contain TscA car
N HG-24r7e 6R8oatIenofe wlfutenhtceeGn roenheGnrAtm olwatghaSae enldenasp tram cr aprice omutumomun___DDuu_ Poonwti._ 1919s54e
--~
APPENDIX C
72-Hour Area Under the Growth Curve Statistics for the Definitive Test
~ -Y
"mPany Sanitzed. Dossnot contain sca cay
GHr-o24w768R:atIeonftfhelGornueGerenoAwlngtcahaSenedles capricorn --~
Dupont11954
72-HOUR AREA UNDER THDEEFGINRIOTWIVTEHTCEUSRTVE STATISTICS FOR THE
GSSRTeOArUTePIrSTSmITCAATeLISnTAINCAiSLY:SFIOSRrAiLccieess?o BPYienk oo .)
dose daosLeval CONT MEW MEDIAN sme swim
21 33
153350,5333 253
3399298800000000 242962400000000
3377362300000000 2399502000000000
137106989783.0860 7216316.85
4s2i11s6ea3ea3es3y0e.no7o's7n
55
1500033 2227218200000000 3281384200000000 2ii70a86d3932s90a59..d097z835 lsLoiemeegesaenesersrnicgyi
SRSHETAPAPOTIRIRTSOT-ICHCRIAEL,MKTEATDMEASOLTNYSOI1FS7:N1L9OiRsMTtAHLUPIRrTSLYDeAOYF A3r1a0s0503
oss EY
sk KET suet puawe
1 ASSL 0.5085 0.17068 0.sdera 0.36761
szowr
--
LRSETEPAVOTERINTST2CI8ROSENTA,TFEMODMRALOANYrSeI1aS7:1L3iFsUTTLHLGPRiHSeDoAeYl 310003
ANALYSIS OF VARIANCE ON FULL Bath S27
street Of LBE pvaur stam
Post 5 oss 0.73288
ATsnhoerasndaaltyasiwsasoffovuanrdiatnocebewinlolrmbaellpyerdfiosrtmreigbuted with equal variances
DNROWESPLEOYSRSTI0S.CRO6EF.N2T5ES,DELO1E2NA.S51T,7A:O215N9, T-H50FU,URLSL1D0A0BYATmAa3/1L00103
BCOTOX OF AQUATIC SELERASTRIM
Obs Class Levels Values
1 ose
6 123436
AOrVeEaR'ALFLROMF-F7UELSLTSDAFTOAR UAsNiOnVgA Weight=NO WEIGHT and Variance Ratioso.33
obs Effect SDaF peSnF Evalue pram
[Sr
son 7.61 0.0020
--
TTT s Company Sanitin zed. Doesnot contain Tsca car
GrH-o2w4t7h6R8tIenoffthleoGurneGeernonAlwgcaaSenldenasruncapricormon
Dupont11954
~
LArSeUaEANFSRO- DFUOLSLE MDAETAANSUsAiDnUgSTWEelDghFeOaRioTHWa1EGFHFTECTaScdovFasSteLoEmNeAStTRoDNro3.3
obTs gLave 50 nsa%s
3358L29o68v00e00n0000
2286226604s99z33..7711
E1S3 13
5iadsooo
249102400000000 27120000
322e86z22666444999333.777111
1133 13
22280000 282645371
AUErSSeITaNIGMTARATOLENPDHFADU=OL.SL0E5DEAFFTOFARECUCTsOSiMnP&gARDIWUSeNOiNNgEShTtY=TNo#OOCRowDmeacnrne.asing
13 AUERWATIVE
WEIGHT and Variance Sationo.3s
Estimate
sIau
sDuimdeesdt pualee
Gmneresoatw
D2DOo0Ss5Ee8 2QT-UE1ADD DOSE 3.1
es
0.0g.1.90090090891653
.o 39280006
3:
DDDo0OSSSEEE
54--11 6-1
*
001954377376 oles
i213e5a3o0000000 Zraocoo
33 3
olo0sss izsoo0s 3
~~
AUErSSeITaNIGNFARATOLENPDHFA2U=0L.5L025DEAFFTOFARECUTCsSOiNnP&gADRWUTeNSiONGNEhSTtToTNo0ORLWoEpDImeGecHarTseaasnidngVaArLiPaEnRcNeATRIaVtEios0.33
XI2aPxRT7_JTI1S5ISJSToAPNN-cDVkAAmLRmUDREIsZEFODsRtJaPOrNzACsRKTAIDECETRaEgsS.TATISTIC
SZPILGLuNRITLFBSRIESASRUPLEVTASTLOUARREETIFFZOO-RRCbODRoeRMcEWrCAeIRaEsDDinPTgSeTApLTWSIRTWHASCIOVNSIIJNeUaIsTrYroCgOrRsRBCRION FACTOR
Jonckieaxe on _TW
Trend Test 3.733Z5o32r
on Dose 0 0.0X00L03I4T3
+
Lowest 5 xeon
Doses thru 100 sine pose
Host
Jonckhoere
Trend
Test
on
Dose
0
+
Lowest
o 4
Doses
thru
` 50
Most,
ar,TE 2.730Z39T6 0.00X87L53T5 xeRorCEstawr bosss
Jonckheaze Trend Test on Dose 0 + Lowest 3 Doses thr 25 Most,
on ore
I
1.705L8O0T5
0.0X5e1L45o2T
xeon: sine bossi
tGJoroon8ucpklhimeneeearaners tsterhsetonudlrdebsubeleftoserxaamtrireneeniddncltfouesdtcehderceiksnuitfcohsre asTnuoemmmrmareryry.eibtioaisbale.
--_
TTT
CompanySaniizeq, Donote oontsain 75a ap:
HGr2o4w7t6h8R:aItnoflfutehneceGraneeGnrAolgwa Saenldnastram capricrnutu
DuPon 11954
--
ECS0 for AREA (UC)
OREBSSPEORNVSEED: &FOPRRMEUDLIAC:TEDfrPonERCoENTACHAT |JESERAPROS N COENTRO"L MEAY
Os CPOzC N we mE cereme sRebeorciG
21 3
50.2050 m0
33 3
338s828s ated
3d8o3u0s4n ers
zoo 3 aml mis
: 7
-0 =
65 w500000 33 2a2m20l8 S2i3ga9sss
3Lp) lag
118 a
ECCoSv0eriuasnicneg NMaetcrhioxd=MaAnRdQUSAtRaDnTd,rdBruEcrevor& VofersPteereagmaWteerigShetreidmeprreosbit
NE. logecx gamma
vo =
glomgeacx vo
200.0.5360538101596
100001.13010353%69
78l21o.l55i56z3s 270o..l5265a01n43
Xodel Parameter Zstinates, 5CS0 and 95% Confidence Bounds
-~
logecx gama
=
sco
472638 163377 40483 6e.ees2 1866 112.661
Model Evaluation essures
SRE ssmR ssroe swom msuams
$967.60 111.608 390.035 6079.21 0.71385
nGfororatmpeihlad2:escrBCiSp0tiBonVs FiInTclBuYdeMAtRhQeUAfRoDlYl,owings
-- - Ts `Company Sanitized. Does not contain TSCA cay
GHr2o4w7s68RaItnefolfuetnhceGorneeGnrAolwgthaSaenldenasrum capricorn PN
Dupont1195
--
APPENDIX D
72-Hour Growth Rate Statistics for the Definitive Test
~~ -
CompanySanitized. Boos notcontain Tera ca;
GHr2o4w7t6h8R:atIenfolfutenhceGorneGenroAwltsh aSnedlenasrum capricorn ~
Dupont 11954
72-HOUR GROWTH RATE STATISTICS FOR THE DEFINITIVE TEST
GSRSRETeOPaUnOTPRIoTSSTiTICAOrmTNnaI,tsSETArDINaCAStLGYniFSnOI1RS7O:G1L_9iPRJsatrtmAeFsIBLtCYEaeDCO samsuo)
dose dEosAeva CONT uEas wenn smomv sms
21 35
32559 333 33
00l.o077s13333 o0l0o7s0e0r
00l.0077a8s9 00100770876
[l.0o00o01i16s23r54r03a33z35s3 Ooisisse:
oo0bo0io0oa7as2si1ae8es8sy0 aoboaran
::
10500 33 00llo067s0s4s3s3 00l.00s7e0s7 1000000280580174577 oo0b0aisaioaeeennsy
SRSHETRPAPOTIRIRTSOT-ICWORINEL,AKTAENTDAELSOTYNSIO1SF7:N1L9OiRsMTtAHLUFIRITSLYDEAOYF 03R10aUvLe03
AQUATIC SELENASTAUN: FULL DATA
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--
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RALYSIS OF VARIANCE ON FULL DAPA' S27
Effect or imme paws sir
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Obs Class Levels values
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Jonckheere Trend Test on Dose 0 + Lowest 5 Doses thru 100 G/L
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Jonckheere Trend Test on Dose 0 + Lowest 3 Doses thru 25 MG/L
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