Document Rpomj1wOrQ1my0ND4zOoO6Dg7
ACUTE TOXICITY TO AQUATIC INVERTEBRATES (E.G.,DAPHNIA)
TEST SUBSTANCE
Identity:Perfluorooctanoiaccid;may alsobe referredtoas PFOA, FC26, or FX-1001. (Octanoicacid,pentadecafluoro-C,AS # 33567-1)
Remarks: The 3M productiolnotnumber was 269. The testsubstance isa whitepowder. The testsample isFC-26 referredtoby thetest laboratorays N2803-3. The purityofthesample was not sufficiently characterizeda,lthoughcurrentinformationindicatesitisa mixtureof96.5 - 100% testsubstance and 0 -3.5% C6, C7, and Cg perfluorohomologue compounds.
Remarks field:The sample preparationdirectiongsivento the laboratory were todissolvethetestmaterialina 50:50 waterisopropanolsolution.In the protocolamendment, itwas statedthatthe sample was combined withisopropanolina 50:50 ratiopriortouse.
METHOD
Method: U.S. EPA-TSCA Guideline797.1300 Test type: Staticacute GLP: Yes Year study conducted: 1995 Year study completed: 1996. Species: Daphnia,magna Analyticalmonitoring:Nominal concentrationsD.O, conductivitpyH,, and -temperaturewere monitored daily. Exposure period: 48-hours Statisticamlethods: LC5o and EC5o values calculatedw,hen possible,by probitanalysis,moving average method or binomialprobabilitwyithnon'lineairnterpolatiuosningthe computer softwareofC.E. Stephan. Test daphnid source: Obtained from culturesmaintainedby T.R. WilburyLaboratoriesInc,Marblehead, MA from an originaclulturefrom AquaticResearch Organisms, Hampton, NH. Test daphnid age atstudy initiation<: 24-hours
Test conditions Dilutionwater: Filteredd,eionizedwater adjustedtoa hardness of 160 mg/L as CaCO3/L Dilutionwater chemistry: Representativesample given inTable 1 ofthefinalreport. Temperature: 19.4- 20.20C
LightingC:ool-whiftleuorescebnutlbswithan intensiotfy58 ft-c. Photopedod of 16-hourslight8,-hoursdark witha 15 minute transitiopneriod. Stock and testsolutionspreparation: A 1,000 mg/L primary stocksolution(50:50mixtureoftestsubstance and isopropanol) was preparedindilutiownater. Aftermixing,the primarystockwas proportionalldyilutedwithdilutiownater to prepare the test concentrations.No insolublematerialwas noted duringthe test. Exposure vessels:300 mL glassbeakers containing250 mL of, testsolution.The approximatedepth oftestsolutionwas 9 cm. Number of replicatest:wo Number of daphnids per replicatet:en Number of concentrations:fiveplusa negativecontrol Water chemistry during the study:
Dissolved-oxygen range (0- 48 hours): 8.7- 8.8mg/L (controelxposure) 8.7 - 8.8 mg/L (1,000mg/L exposure conc.)
Conductivityrange (0- 48 hours) 590 - 610 pmhos/cm (controelxposure) 570 - 590 pmhos/cm (1,000mg/L exposure conc.)
pH range (0- 48 hours) 7.9- 8.4 (controelxposure) 7.5- 8.0 (1,000mg/L exposure conc.)
Test temperature range (0- 48 hours) 19.5- 20.20C (controelxposure) 19.4- 20.2 C (1,000mg/L exposure conc.)
Element basis: mortalitaynd immobilization
RESULTS
Nominal concentrations: Blank control1,30, 200, 360, 600, 1,000 mg/L (Tested as a 50:50 mixtureoftestsubstance and isopropanol).The concentrationoftestsubstance insolutionwas 65, 110, 180, 300 and 500 mg/L
Element
value (as-tested): 24-hour EC5o 24-hourLC50 48-hour ECso 48-hour LC5o 48-hour NOEC
840 (740- 970)mg/L 1,000 (600->1,000)mg/L 720 (660- 780) mg/L 800 (700 - 920) mg/L
= 360 mg/L
Element value (based on concentration oftestsubstance In solution):
24-hour EC5o = 420 (370 490) mg/L
24-hour LC5o 48-hour EC5o 48-hourLC50 48-hour NOEC
500 (300- >500) mg/L 360 (300- 500) mg/L 400 (350- 460) mg/L = 180 mg/L
Allelement valuesbased on nominalconcentrations.
A toxicittyestconducted previouslywithisopropanolat390 mg/L showed no mortalitoyrsublethaleffects.
StatisticaElvaluation:The LC5o and EC5o valuesand 95% confidence intervalwsere calculatedwhen possibleby probitanalysis,the moving average method or binomialprobabilitwyithnon-linearinterpolatiounsing the computer softwareofC.E. Stephan.
Biologicalobservations: Daphnids inthe controlt,he 130,200, and 360 mg/L treatmentsappeared healthyand normal throughoutthe testwith no mortalityi,mmobilityor overtclinicaslignsof toxicity.
Cumulative percent mortality:
NominadAs-Tested
Conc.,mg/L
24-hours
48-hours
Control
0
0
130
0
0
200
0
0
360
0
0
600
1
10
1
15
1,000
5D
80
Control response: satisfactory
CONCLUSIONS
The as-testedtestsubstance 48-hourLC5o forDaphnia magna was determined tobe 800 mg/L witha 95% confidenceintervaolf700 - 920 mg/L. The as-testedtestsubstance48-hour EC5o forDaphnia magna was determined tobe 720 mg/L witha 95% confidenceintervaolf 660 - 780 mg/L. The as-testedtestsubstance 48-hour no observed effect concentration(NOEC) was 360 mg/L. The 48-hour LC5o forDaphnia magna based on the concentrationoftestsubstance insolutionwas determinedto be 400 mg/L witha 95% confidenceintervaolf350 - 460 mg/L. The 48-hour EC5o forDaphnia magna based on the concentration oftestsubstance insolutionwas determinedtobe 360 mg/L witha 95%
confidenceintervaolf300 - 500 mg/L. The 48-hourno observed effect concentratio(nNOEC) forthetestsubstanceinsolutiownas 180 mg/L.
Submifter:3M Company, EnvironmentalLaboratoryP,.O.Box 33331, St. Paul,Minnesota,55133
DATA QUALITY
ReliabilityK:limischranking3. The studylacksanalyticamleasurement oftestsubstanceconcentrationisnthetestsolutionasnd sample purityis notsufficientclhyaracterizedA.dditionalltyh,ereappears to be a discrepancybetween thesample preparatiodnirectiongsiventothe laboratoraynd the procedureconductedby thelaboratortyopreparethe testsolutions.
REFERENCES
Thisstudywas conductedatT.R.WilburyLaboratoriesI,nc.,Marblehead, MA, attherequestofthe3M Company.
OTHER
Last changed: 5/25/00
Study Titl*
A--ut* Toxicity of N2803-3 to the Daphnid, DAphnim magna
Data Rwquire@nt: Guidelina Number U.S. EPA-TSC-A, Guideline 797.13CO
Authors iT@mothy J. Ward
eanne r. Magazu Robert L. 3oeri
Study Cowplatckd March 7, 1996
Sponsor
3K Company
Environmental. i.aboratory
935 Bush Avenue
Building 2-3E-C9
Vk
St. Paul, Minnesota 55144
Testing Facility
R"'@Wilk@@
'"Laboratories, I
--y-@Oaks Lane
headthassachusetts
C1945
T. R.' wilbury, S udy@Number i'@2
Page
f
ItA
I. GWD IJLBORATORY
PX4CTICZ
COMPLXA*CA
STATDCZXT
This study was conducted according to EPA Good Lat>orator-y Practic;
R*grulations (40 CirR 792), the test substance was
Laboral,ories. Deviations
with the assumed
from the
following exception. The stability
but not verified at T.R. Wilbaray
study protocol &,-idprctc>ccl amendw@.-its
a-re Presented in Swction XII of this report.
:ea,hr.eP. ma3aiu: study Director
R.
u
9
.*6z Pm'ge i or 25
11- QVJLLITT A.SSUR"CS
STATZKXWT
Submitted byt
T.R. Wilbur-y Laboratories, 40 Doaks Lane marbleh*ad, HAssachusetto
Inc.
@-ortifi,:ation:
.R. wilbur-Y Laboratories, Inc. study number 892-TH, Acute -oxicity of N2803-3 to the Daphnid, Daphnis magra, was audited by the Quality Assurance unit for compliance with the protocol, standard c)pmrating prc>ceduroo, and
appi icable Good Laborator-y Pract'@ces (U.S. EPA, 1993) . QU&Iitv assurance audits were per-ormad and the findings rep<)zted to T.R. Wilbur-y Laboratories maragement and the
st,,;dvdirector on the following dates:
Aspect Audi.,@ed
Protocol@ ir.-life:
Draft Report: F'@nal Report:
Audit Date
B/ 4 ','95 9/ @1/95 9/ 6/95 1/ 2/96 3/ 7/96
Reported to Study Director
B/ 4195 9/ 1/95 9/ 6/95 1/ 3/96 3/ 7/96
Reporteil to
Managemeiit
B/ 4/9'i 3/ 7/91! 3/ 7/91; 3/ 7/91i 3/ 7/9@i
Arthur P.-;Faradic@'
T.R: wilbury'study Numb*r 892 -TH
-4 .'age 3 of' 2:)
111, TABLX or cow.-IDrTs
SECTIONs
1.
Cood Lab-orator-y Practice compliance statement
ii. Quality Assurance Statement
Ill. Table of Contents
IV. Index Of Tables and Figures
V.
SLnmar-y
Vi. General information
VII. Introduction
VIII. Methods and Materials
IX. Results
X.
References
Xi. Signature Page
XII. Protocol Amendments and Deviations
Appendix A. Water Quality Data from Toxicity Test
F)kGg 2 3 4 5
7 8
13
19 20 21
@k"
-Study Number 892-TH
'A,
IV. INDBX OF TJLBL43 AXD rlruvjg
Chemical characterization of a representative sample of dilution water used for the toxicity test with daphnide, DaFhnia migna, and N2803-3.
Table 2. .able 3.
Test system ou=ar-y for the toxicity test with N2BO3-3 and daphnide, --aphniamagna.
su--vivaland sublethal effect data from the toxicity test with ciaphnide, Daphnis "gna, and N2 8 0 3 -3
Table 4.
Survival and sublethal toxicity test with
isopropanol.
effect data from the Da.z@hniama7na, ard
Table 5.
median lethal ccncentrations (Lc5Os) anJ =edian
effecti,,e concentrations (EC50m) from t@,@etoxicity test with daphnids, Dap.@nia magna, and
Table A.I.
Disso@ved oxygen concentration, conductivity, pH, and te=Deratare measured during tie t,@x:@;.tytest with daphnids, Daphn@a magna, and N2803-3.
Table A.2.
Dissolved ox,.igen concentration, conductivity, pu, and temperature measured during the toxicity test with daphnids, Daphnis migna, and isopropanol.
F4gure 1.
Survival and sublethal effects of dapt)n.@,la,E;aph.,Iia magna, exposed to N2803-3 for 48 hours.
PKGE 9
ti 4 5 17 22 24 16
T. R.' Wi lbury
study
Namber
892-TH
-k
page 5 Of,25
V. summ"y
The acute toxicity of N2803-3 to the daphnid, Daphnis magn&, is
described In this final report. The test was conducted for 3m coinpany for
48 hours from August 30 to Septerl>er 1, 1995, at T.R. Wilbuz-y
Lat>oratori*s, Inc. in Marblehead, Massachusetts.
It was conducted by
isanne Magazu, Peter Yowalski, Jacqualir.9 Nevius, Kaven Stevens, Peter
"vium, Robert Bo*ri, and Timothy wa-rd according to the protocol developed
for -,.R. wi!bliry sponsor.
S@udy
N@,imJ:>e8r92-TH.
N2803-3 was supplied by the
7!'.. test was performed
under static conditions with five
of test s,;t)atanceand a dilution water co,,itrolat a te,T@per.tur. f 1.9.4-20.2'c. The dilution water was filtered doionized
water cc,;Ilected--etMarblehead, Mazaachuset:ta and adi,@ated to a hardness of
160 mg@@L as C&Co,. Aeration was not required to maintain dissolved oxygen
concentrations &nova an acceptable level.
The sample of N2803-3 supplied by the sper)oc.,,@aB co=,@bined with
is,)Propanul in a 50:50 ratio prior to use (a toxici!iv test. was conducted
wit.li39@ mg/L is,@p.-opanol and no idort.ality or aut,le*.Iialeffects e,e
ob5arvod).
No,.nina I concentrations
of N2803-3
as-roceived were 0
65, 110, 180, 300, and 500 mg/L. Daphi@ da uoed in the test
were less than 24 hours old at the start of the test.' They were produced
at T.R. Wilbury i.aboratories and acclimated under test conditions for
their entire life (cultures that produced test specimens were maintained
at test condit i-ons for more than 7 days). After 48 hc)urs of exposure co:itro' ddphn ds had an average wet weight (blo',ted) of 0.53 mg. dal)hni2s were in good condition at the beginning of the study.
the All
Fxposure of daphnide to N2BO3-3 resulted in a 24 hour median loth!Ll
cor.contrati.in (LCSO) of 500 mg/L N2803-3, a 48 @iour LC50 of 400 mg@L
N2803-1, a 24 hour median effective concentration (EC50) of 420 mg/L, aid
a 48 hour ECSO of 360 mg/L N2803-3, based the concentration of N2803-3 in the test Bubstance. The 48 hour no observed effect -aricentration (NoE,-)
is 180 mg,!L, based or, the concentration The 24 hour LC50 is 1,000 mg/L, the 48
EC50 is 840 mg/L, the 48 hour EC50 is 360 mg,/L, based on test substance inc@p ropanol).
of N2803-3 ip. the test substanc@3.
hour LC50 is 8- '0mg/L, the 24 hour 720 --Ig/L,and the 48 hour,NOEC Ls. as-tested (50% N2803-3 an d 51)11,'4'
T.R. Wilbury study Number 892-TH ire
Page-6'of
VI. GE"RAL I"ORXATIDK
Material T*Stedj Sponsort
Hterial Shipped Fromi
Reported Purity:
study init@ation Date:
ExPelimental Start Date:
Exper@@Mental Date:
Completion
Study Completion Date:
Azchive Lc>cation for tt,@eRaw Data and a Copy of t@.0 Final Report:
N2803-3
3m company Environmental Lab-oratory 935 Bush Avenue Building 2-3E-09 St. Paul, Minnesota 55144
Im company Environmental Laborator-y 935 Rush Avenue Building 2-3E-09 St. P;ul, Minnesota 55144
100%
August 16, 1995 August 3D, 1995
signed)
October 6, 1995 March 7, 1-996
T.R. wilbury Lab<)ratcries, Inc. marhlehead, Massact,.usetts
T.R. wilb
.,Stu @N
$92-TH
ago- o 2 5:
vii, INTRODUCTION
This study was sponsored by 3m company, st Paul, Minnesota, Th. objective of the study was to determine the 24 aud 48 hc>ur median lethal concentrations (LC509) and modian effective concentrations (EC502) of the test substance to the daphnido Daphnis magns, a freshwater invertebrate, ur,@ierotati-- conditions. Th* test protocol was based on U,S. EPA method, (U.S. EPA, 1993). The report contains sections that describe the 7@ethods and RL&terialm employed in the study, and results of the investigation. T,Iiareport also contains an appendix tha* presents water clualit:ydata collectad during the test.
TEST SUBSTAJZCE:
VIII. ME"ODS AND MATZRIALS
-@he sa-@p,e of N.'803-3 (-@.R. Wilbury Laboratories Sample Number 52'.)
was delivered to T.R. wilbury Laboratories on August 14,
cardboard box at ambient temperature.
1995 1,1 a
It was contained in an
app@o::izr,atoly50 MI glass )ar. The ',&be! attached tc)the bottle inclu@,led
the following information; "N2eo3-3'.
N2903-3 (a white solid) was supplied by 3M company, S. Pau.,
Minnesota.
Prior to use the N2803-3 was stored -.,ithe dark at rooz,.
temperature. The sample of N2803-3 supplied by the sponsor was combined
with isopropan.-l in a 50:50 ratio prior to use. This 50:50 mixture was
then considered to be 100% test substance during the performance of the
toxicity test but all results are reported on an active ingredient: basis
(active ingredient
is the test substance
as-received
minus
the
is,,.)propanol). The test substance was aaau=ed to have a purity of 100%
active ingred-ient and to be stable under storage and testing conditions.
Unused N2903 3 will be returned to the sponsor.
DILJJTION WATERI
water used for acclimation of test organisms and for ali toxicity
testing was deionized water collected at T.R. Wilbury Laboratories i,@i
Marblehead, MasBachuse,ts.
Water was adjusted to a hardness of 160-
130 =g/L an CACO and stored in 500-gallon polyethylene tariks where it was
aerated and continuously passed through a particle filter, ultraviolet
ste--ilizer, and activated carbon. Results of chemical,,'analysis of a' ,,representative sample of water are presented in Table'l.,'
TZST ORC-AMISMI
@,";ij'uveniledaphnids,
e loyed.as,ztentiorganiams
1.@@sa' tha@n 24'hours'old at the start of,the test.,'@Tboy .w'or"*4'from a mi ere
r7@source'@-ar@d'!.'@s re @'@identified
ing.,an appr6priii a
"specixb@ns'"re produced by a duslt d@phnidx'that'ver ma@i@ta nod undoi.test @,4@ conditions at the T.R. wilbur-y facility for more than 7.,days.
14'c V-@v
T.R. wilbury study Number 892-TH
Page 8 of 25
4
Table 1
Chemical characterization of a representative S=ple of
di'.utio,,wiater used for the toxicity test with daphnids, Daphnis magna, and N2803-3.
P&ramet*rl
unit of Measurazent
Detection Limit
Meao,,ired value
Metals Aluminum kr ae,.Iic! Be@-cn cadmium chromium ccba'@,:Copper 'r:)n Lead Mercury Nickel silver z i r.c
Nit.rate Chloride Fluoride Total Organic Carbon Total Phosphorus
mg/L mg/L mg/L mg/L Mg/L ing/L mg/L mg/L mg/L m(g/L Mg/L mg/L mg/L
mg/'- as 4 mg/L mg/L
mq/L mg/L
0.1 0.01 0.5 0.0002 0.01 0.03 0.005 0.03 0.005 0.0003 0.03 0.02 0.02
0.05 I 0.1
I 0.03
NE)' ND ND ND ND ND ND 0. 03 ND ND ND ND ND
N
ND
ND NL,
organc,chlori:ie
Pesticides
pg/L
0.5
ND
Toxap@iene
pg/L
2
ND
orgaiophosphor,,js
Pesticides
;jg/L
0.5
NO
Dimet.'@cate
pg/,-
2.0
ND
TEPP
;jg/L
2. C
ND
14onc>crotophoo
pg/L
20
ND
PCBN
0.5
ND
ji
Notes:.,?;,
Parazaters
dilution
analyzed
testing.
ND
not
we re measured in do onized water@'(used to formulate'
water) that@.wankFollected.,
du--ing -AUcjust;'@ 1995 and
by Pacip't@l@6 as. p
6,4roUtin!l,@a
or,@quality',-
doteczoa4
I>o
T, R. Wilbur-y Study Numb-or 892-TH
i@@
Page 9 o@ 25
The original culture was obtained on June 1, 1994 from Aqujktic Research Organisms, Hampton, NOW Hampshire. Keasurem*nts made during the 7 days before the start of the test with N2803-3 indicated that the culture temperature ranged from 19.5 to 20.5*c and the dissolved oxygen concentration was always at least 9.1 mg/L and measuremorts made during the 7 days before the start of the test with isopropanol indicated that the culture temperature ranged from 20.1 to 20.9'C and the dissolved oxygen concentration was always at least 8.3 mg/L- The cultures each produced young prior to day 12 and a ouboample of adults produced at least 3 young 'daphnid/day for 7 days before the start of the tests.
During acclimation da.,hnids war& not treated for disease, they were free of apparent sickness, inj,-,ries,and abnormalities at the beginning of the test and there was no mortality during the 48 hours preceding the start of the test. The culture was sup,-lied with a yeast/trout chow mix
DC-24 and the freshwater alga, Selanistr-um capricornutum, dailv during acclimation. Daphnidz were not fed during the test, Contr.i daphnids, blotted and weighed at the conclusion of the toxicity test with EL2803-3, ave.-aged 0.53 mg and the loading -ate durinj the definitive toxicity test was approximately 10.4 mg/L and 40 daphnide/L.
TOXICITY TESTING:
A static screening test was conducted frc,m Aug,-sl 23 to 25, 1995
.ith five concentrations of test substance and a dilution water control.
Nomina. ccncentratiois of N2803-3 were 0.050, 0.50, 5.0, 50, and 500 mg/L.
After 43 hours of exposure there was at least 95% Bur-vival in the control
and at 0,Cc.0, 0.50, 5.0, and 50 mg/i, and 15% survival at 500 mg/L
(surviving daphn@ds exposed to 500 mg/L were i=.cb@-lized).
-
@he def@.nit4ve toxicity test was perfoi-med from August 30 to
September 1, 1995, according to T.R. wil-bury P--otccol 892-TH (Acute Toxicity of N2803-3 To The Daphnid, Daphnia magna). The protocol, which
was signed by the study (iirector on August 16, 1995, is based on
proced4res of the u.s. Environmental Protection Agency (1993). A summary of the test system is presented in Table 2.
The test was conducted at a target temperature of 20 1 1*c with f ive
concentrations of test substance and a dilution water control. Nominal@a@"
concentrations of the test substance were: 0 mg/L (control), 130 200t'
360, 600, and 1,000 mg/L (concentrations of N2803-3 were 65, 11 , 306""
500 mg/L) . A 1,000 mg/L (5DO mg/L N2a03-3) stock solution was
a .1
by combining 2. 0 g of " test 'substance (50: 50 mixture of N2
3n
isopropanol)
and dilution @@tatera n, a @ Class A,, Volumetric
f 1 a'B* an
a d Usti g.the.final at Gck,a-'ol"ut'i'o;'in@'@'t. 2,000 .1 @i.,Thestock j. , @i@n
v@Was mixed
and,,then
appropr
atea=ouni'@s',ioiD
this stock' - so I u-tion wer
dd;d
1
to dilut i on water' -1-in,,test.,vesBelal-tcv@!ior"mulate@
ite-st' 'm'odfa-@"-w-ithl*th'
a's o
so 1 ve n t
T.R. Wilbury Study N er,
,,i.age
C)f
Table
Test system summary for the toxicity test with H2803-3 and daphnide, Dephnim mAgnA.
Species: Ag&/SIZO Of Test organisms: Too' D'@rationi Teat vessels: Test Media vcl,=e: DePt@',of Test media:
of Repl4@cate -eat Vessels: Or;anis-.isper Replicate: TO-P-rature Range: Miriir.,@Dmissolved oxygen:
Lignt Intensity: Water Quality measurements:
Acceptability Criteriat@@.'
DaphniA magn&
<24 hours old
48 hours
300 ml glass beakers
250 inl
9 cm
2
io
19.4-20.2*c 8.7 mg/L
16 hours light and 8 holizadark, 58 footcaidles
Dissolved oxygen, PH, temperature, and conductivity daily in each test vessel. Hardness, total organic
ca-rb,on,and alkalinity once in the control. Residual chlorine daily Dilution water analysis is au=aried in Table 1.
24 and 48 hour LC50 and EC50 (based on
bilization),48 hour no observed
effect level@.@,,, .11, t".-
-@@ . , "'
SIOS control mor@tality temps ratlire,rang*.
-f T. R. wilbury study NuaLber y -'EH
of-'25,4
Twenty daphnide were indiscriminately and *qua',ly distributed among two replicates of each treatment. The text was performed in 300 ml glass beakers that contained 250 ml of test solution @water depth was approxiinate.y 9 cm). Test vessels were randomly arranged in an incubator d,iring the 48 hour test (a random numbers table was used to select the location of each vessel) and they were loosely covered.
A 6 hour light and 8 hour dark photoporil)d with a 15 minute transition period was automatcally maintained with cc>ol-whitsfluorescent lights that provided a light intensity of 59 fc>otca,-idlesA.eration was not required to maintain dissolved oxygen concentrations above acceptable levels and daphnids were not fel during the test.
The nu=er of surviving organisms and the Occurrence of sublethal effects (i--=cbilization, lose of equilibrium, loss c @, reflex, ex,,i,,&L,ilityd,.@.scoloration, or change in t>ehavior) were dater-mined vis@@ally Id recorded initially and after 24 and 48 ho,4re. Dead test ctQar.@s:r.wsee removed when first observed. Dissolved oxvgeli (YSI Model 57 meter; instrumert number 1), pH (Sezkman mode@ pill 12 meter; instrument number 124), conductivity (Cole Pa--mermeter, instrument number 94), and ta:zperature @Bockinanmodel pHl 12 mate:; instrument n=ber 124, probe ph@4) ozo measured and recorded daily ir.each test chamber that containid l@@-a animals. The temperature in a beaker of water l@ncubated among t.io test veas4a!awas recorded continuously during the tejt.
A static test was conducted from October 4 to 6, 1995 with 390 mg/L is@.p--z)pano(l@1.125ml isopropanol brought up to 2@O ml with dilution wate:) and a diluticn water control. The test was established with 2 replicates of each concentration and 10 daphnids per replicate and it was cond,-@cteid,-a. ma.,i,-siiemrilar to the test With N2803-3. 'rest ,easels were randomly arranged in a water bath and the light intensity was 13 to 25 footcandle5.
STATISTICA.L METRODST
Results of the toxicity test were interpreted by standard s,at4stical techriques. computer methods (stephan, 1993) were used to cal.--luaite LC5,'s and EC50s. All calculations were performed by using now-i,-iaclonce,-,t:ratioposf both the test substance (as tested) and N2803-3
(as received) and the number of-live or unaffected daphnids.
7
oboe--ved effect concentration is the highest
The nto
substance
con@:entration of te
effects. that did not cause any toxicant-related mortality or sublethaal
T.R. wilbur-y study Number 892-TH
rt Page 12 of-25
ie
11- RZSVLTS
NO insoluble material was noted during the
water Table
quality data generated by the &cuts toxicity 3 and Appendix A, respectively. During the
measured concentrations of dissolved oxygen ranged
test. test
test from
Biological and a-represented in
With N2803-3 the 8.7 to 8.8 mg/L,
the temperature ranged from 19.4 to 20.7*C, conductivity rknged from 570
to 610 wmhon/cm, and pH ranged from 8.3 to 8.5. During the test with
isoptopanol the measured concentrations of dissolved oxygen ranged from
8.1 to 8.5 mg/L, the temperaturt ranged from 20.4 to 20.9*C, conductivity
ran-,eL' from 530 to 540 ;jmhca/cm, and pH ranged from 8.3 to B.S. 0..
hu;-@,@zedpercent survival occurred ii the control exposures and these
daphnids did not exhibit any sublethal effects. During the toxicity test
with 3RO mg/L i-&Dpropanol no mortality or sublethal effects were oboe-,.,ad
t-e 48 hour EC50 and LC50 values are greater than 390 mg/L (Table 4).
The 24 and 48 hour LC50s and EC50a for daphn@@ds exposed to N2803-3 axe @@.e!se,-.tedin Table 5.
Exposure of daphn'.da to N2803-3 resulted in a
24 ho@r -iedian lethal concentration (LC50) of 500 mg/r. N2BO3-3, a 48 houz-
LC@' of 400 mg/L N2803-3, a 24 hour median effective concentration (EC50)
of 420 mg/L, and a 48 hour EC50 of 360 mg!L N2803-3, based on the N2803-3
ccr,.ce:i@.raticriin the test substance.
The 48 hour no observed effect
ccncentration (NOEC) is 180 mg/L, based on the N2803-3 concentration in
the test substance.
BCO mq
-he 24 hour LC50 in 1,000 mg/L, the 48 hour LC50 is
'L, the 24 hour EC50 is 840 mg/L, the 48 hour EC50 is 720 mg/L, and
the 48' hour NOEC is 360 mg/L, based on test substance
N29@,3-3 and 50% isoprz@panol).
as-tested (50%
The slope of the 48 hour dose reopcnbe
curve (based on mortality and ixmobilization) is 10.5.
T, i@.T.. R wilbury Study Number 892-TH
f 25
Tabl* 3.
Survival and sublethal effect data from the toxicity test with daphnide, Daphnis magria,and N2803-3.
Nominal. Concentration
of N2803-3 (mg/L)
Rep.
Number &live
--
0
24
48
hr
hr
hr
0 (control)
1
10
10
10
2
1)
10
10
65
1
10
10
10
2
10
10
10
iio
1
10
10
10
2
10
10
10
180
1
10
10
10
2
10
10
ic
3co
1
10
9
8
2
10
9
9
500
1
10
5
2
2
10
5
2
Note: Affected daphnide were i bilized.
Number affected
0
24
48
hr
hr
hr
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
3
2
0
2
2
,T.R. Wilbu ry Study Number SYI-TM@
W'7'
page 14 or 5 1 14
Table 4.
Survival and sublethal effect data from the toxicity test with daphnide, Daphnis magna, and isopropanol,
Nominal Concentration of isopropanal
(r,g/L)
0 (control)
39.0
Rep.
2 1 2
Number alive
0
24
48
hr
hr
hr
10
10
10
10
10
10
10
ID
10
10
10
10
Numl@.*r affected -
0
24
48
hr
hr
hr
0
0
0
D
0
0
0
0
0
0
0
0
R.,Wilbury Stucly Number,892
%
20
1 5 C,
0 65
-
1 10
180
300
500
Ccnc,i-l I
T
DoPhnids
UnOffected DOPhr@ds
gure '1* urvival and aLLblethal effects of daphnids D,
na,
At,;
OXPOM@D to N2803-3 for 4B ho urn.
'S't
wi]_bL,@@Siudy'N uxlb*r 892-TH
Table 5.
Hodian lethal concentrations (LC50s, and median effective
concentrations (EC505) from the tOxicitY test with daphnids, Daphnis magna, and N2803-3.
Exposure period
LC50/E@-50
95 percent confidence limit
calculation method
Calculated based on Test Substance as-T*sted
LC50
24 hourfi
I,OCC mg/L
600 to
mg/L
43 hours
800 mg/L
700 to 920 mg/L
Binom-ia!/Nonlinear Interpolation
Probit
24 hours 48 hours
840 mg/L 720 mg/L
740 to 970 mg/L 660 to 780 mg/L
Probit Probit
Calculated based on the x2803-3 concentration in the Test Substance
24 hours
500 mg/L
300 to >500 mg/L
Binomial/Nonlinea-r
48 hours
400 mg/L
350 to 460 mg/L
Probit
EC50 24 hours 48 hour
420 mg/L 366"'Mg/L,I,@@'
37 0 to 490 mg/L P @'F
3DO tO'500 Ing/L'
Probit
Binomial/Non Interpol&
Note: The 2 4 an d 48 hour,.Ec an LC50,va
-,L, soprop
grea
tha"n@'4'-'
fromili'b@'t@xii ty.test,'with
wilbu Stu-clyNumber_
a o 17:of
<7
Rsyzxjmtczs
ASTM.
1986. Fishes, Book of
Standard Practice Macroinvertabrates, standards.
for conducting Acute Toxicity and Amphibians. E-729 -SO.
Too th
Atnsnuw!
StoPhan, C.E. 1983. Computer Program for the calculat4
U.S. EPA. Duluth, MN. Personal co=unication.
of LC50 Values. "4@ 'on
L'.5, EPA. 1993. TOX'C Substa.-,Ces Control
Ru'es.
section 797.1300.
Act Test Guidelines,,
Final
U.-.
EPA. 1993,
40 CFR Part 79@'. Toxic
Cc@od Laboratory
Practice
Title 4), @'uly 1, 1993.
Standards.
substances Code of
control Federal
Act (TSCA) Regulation
t 4@
zg
wil-bu
fs;tu N
-TH 2
of-2
r
Zi.
I'GKATun
PAGE
study
P. Kagaiu Dire ctor
Aquatic Toxicologist
Peter L. Kowalski Aquatic Toxicologist
,B@l@oclqotgiel@Ine M. ist
NeVius
Raven m. Biologist
tor:l POter Biologist
C. Noyia.
Ro
rt L.
Coauthor
Boeri
Tl*othy
of.i
d
.7 T
lbu
stud
N
r-"'892 -TH
25,
PROTOCOL AXKXDKDffg AND DlgvlATIOuS
PROTOCOL AXENDOCKNTS:
The protocol was amended to specify the nominal concentrations for the definitive test, to indicate that the test substance was combined in a 50.50 ratio with isopropanol, and to note that a toxicity test would also be conducted with a single concentration of isopropanol.
PROTOCOL DLIVIATfONSt
The nominal ".aopropanol cc,-icentration wa5 390 mg/-- rather than 500 mg/L. This deviation did not affect the outcome of the test and no other deviations were made from the protocol.
T.R.,Wilbury Stud y Number 892-TH
age 0 of,'25
APpendiX Al WATER QUALITY DATA, RROM TOXICITY @KSL
tfv, T.It.'WilbuX-y B tudy Numi>er 892-TH,
@e-
'of 25
C4
Tabl* A. 1,
Dissolved
oxygen
concentration,
conductivity,
tfmporatur* measured during the toxicity test with
Daphnia Magna, and N2803-3.
PH,
and
daphnide,
Nominal Concentration
of M2803-3 (m@g/L) 0 (control) 65
53)
Dissolved oxygen (mg/L) Conductivity (pxbog/cm)
0
24
48
Rep@
hr
hr
hr
0
24
48
hr
hr
hr
1
8.7
8.8
9.7
2
8.7
8.8
8.7
1
8.7
8.8
8.7
2
8.7
ele
8.7
1
8.7
8.8
8.7
2
8.7
8.8
8.7
8.1
8.8
8.7
2
8.7
8.8
8.7
8.7
8.8
8.7
8.7
8.8
8.7
8.7
8.8
8.7
2
8.7
8.8
8.7
590 590
590 590
590 590
590 590
580 580
570 570
600 600
600 600
600 600
600 600
590 590
590 590
610 610
610 610
6io 610
600 600
600 600
590 590
44 f
T.R wi
t ud y
uml>er
@2-TB'7--
Ilk Page
17
Table A, 1, Continued.
Ncminal Concentzation
Of N2803-3 (Mg/L)
0 (control) 65
180
500
PH
0
24
48
Rep.
hr
hz
hr
1
8.0
7.9
8.4
2
8.0
7.9
8.4
1
7.8
7.9
8.4
2
7.8
7.9
8.4
1
7.7
7.8
8.4
2
7.7
7.8
8.4
1
7.6
7.7
8.3
2 7.6 8.3
1
7.6
7@7
8.2
2
7.6
7.7
811,
1
7.5
7.6
8.0
2
7.5
7.6
7.9
Temperature (-C)
0
24
48
hr
hr
hr
20.2 20.2
20.2 20.2
20.2 20.2
20.1 20.1
20.1 20.1
20.2 20.2
19.6 19.6
19.5 19.6
1.9,6 19.5
29.6 19.5
19.6 19.6
19.6 19.5
19.5 19.5
19.5 19.5
19.6 19.6
19.5 19.4
19.4 19.4
19.4 19.4
udy Number 892-TH
Table A. 2.
Dissolved
oxygen
concentration,
conductivity,
te"r&tqro measured during the toxicity test with D&phniA niagna, and isopropanol.
PH,
and
daphnids,
Nominal Concentration of lscpropanol
fmg/L)
0 (control)
390
Rep.
Dissolved oxygen
-
-
0
24
hr
hr
(IDg/L)
48 hr
CondQctivity (wmhon/cm)
0
24
48
hr
hr
hr
1
8.1
8.5
6.4
2
8.1
8.5
8.4'
1
8.1
8.5
8.4
2
8.1
8.5
8.4
540
530
530
540
530
530
540
530
530
540
530
530
ft T.R. wilbury study Number,c7.e-TH"',,-
p age 24
25
i@4
Table A@2, continued.
Nominal Concentration of loopropanol
(Mg/L)
Rep.
0 ico:,trol)
1
2
390
1
2
PH
0
24
48
hr
hr
hr
.5
8.4
8.3
8.5
8.4
0.3
8.5
8.4
8,3
9.5
8.4
8.3
Temperature (1c)
0
24
48
hr
h
---------------
20.4 20.4
20.9 20.8
20.6 20.6
20.4 20.4
20.8 20.9
20@6 20.6
T
T.R. wilbu
Number 892-TH
-Page 25 of .2