Document 6bGdrOZo3gQj8zD5dKNkKmDxE
ACUTE TOXICITY TO AQUATIC INVERTABRATES TEST SUBSTANCE
(DAPHNIA MAGNA)
Identity:Perfluorooctanoiaccid,ammonium salt;may alsobe referredto as PFOA ammonium salt,Ammonium perfluorooctanoatPeF,O, FC116,FC-126, FC-169, FC-143 or as the major component of FC1015. (Octanoicacid,pentadecafluoroa-m,monium saltC,AS 3825-26-1)
Remarks: The testsample isFC-1015. It'psuritwyas notsufficiently characterizedt,hough currentinformatioinndicateissa 30% straighctarbon chainversionof FC-143-in80% water.The 3M productlotnumber was "HOGE 205".
The followingsummary applies to the testsample as a mixture of the testsubstance in water solutionwith Incompletely characterized concentrationsof Impurities.Data may not accurately relatetoxicityof the testsample with thatof the testsubstance. Data was used to compare toxicityof the branchedlstraightchain ammonium perfluorooctanoatehomologue mixture In FC-143 vs. what Is supposed to be the 100*/ostraightcarbon chain ammonium perfluorooctanoate In FC-1015.
METHOD:
Method: USEPA - TSCA, Guideline797.1300 Type: Staticacute
GLP: No
Year completed:1996
Species:Daphniamagna
Supplier:In-houseculturesI.nitibarloodstockfromAquaticB!oSystems,
FortCollinsC,olorado
Analyticalmonitoring:Temperature,pH, conductivitaynd DO
Exposure period:48-hours
Test organism age: <24 hours old
Statisticamlethod: Probitanalysis
Test conditions:
Dilutionwater: Deionizedwateradjustedto a hardness of 160-180
mg/L (asCaC03) and passed througha particulaftielteurl,traviolet sterilizearn,d activatedcarbon.
Dilutionwater chemistry:
Hardness: Alkalinity: TOC:
176 mg/L as CaC03
108 mg/L as CaC03 <1 mg/L
Lighting:Cool-whitefluorescenltightast 130 foot-candlesA. daily
photoperiodof16 hours lighatnd 8 hours darkwitha 15 minutetransition periodwas maintainedthroughoutthetestingperiod.
StockandtestsolutiopnreparatioTne:stsolutiownesrecreatebdydirect weightsaddition. Exposure vessels: 300 mL glassbeakerscontaining250 mL testsolution(9 cm depth). Number ofreplicatest:wo Number ofdaphnids per replicate1:0 Number ofconcentrations:fiveplusa blankcontrol Water chemistryduringthe study:
Conductivityrange: (0-48hours) 610 - 620 pmhos/cm (controelxposure) 710 - 720 pmhos/cm (2000mg/L exposure)*
pH range (0-48hours): 8.2- 8.3(controelxposure) 8.1-8.2(2000mg/L exposure)*
Temperatur'e range (0-48hours): 20.4- 20.7OC (controelxposure) 20.3- 20.8-C (2000mg/L exposure)*
Dissolvedoxygen range (0-48hours): 8.1- 9.1mg/L (controelxposure) 8.1- 9.1mg/L (2000mg/L exposure)*
2000 mg/L (secondhighestconcentratione)xposurevaluesused because thehighestconcentratiounsed resultedintotalmortalitbyy 48 hours.
RESULTS
Element values:
24-hourEC50 = 1790 (1550 -2070) mg/L 48-hourEC50 =1200 (730 -2000) mg/L 48-hourNOEC = 730 mg/L
Element valuesbased on nominalconcentrations
Remarks: Testingwas conductedon themixtureofthetestsubstance as describedinthetestsubstanceremarksfieldT.he valuesreportedapplyto thatmixtureand notthetestsubstance.
CONCLUSIONS
The FC-1 015 48-hour EC50 was determinedtobe 1200 mg/L witha 95% confidenceIntervaolf730 to2070 mg/L. The 48-hourNo Observed Effect Concentrationwas 730 mg/L
Submitter: 3M Company, EnvironmentalLaboratoryP,.O. Box 33331, St. Paul,Minnesota,55133
DATA QUALITY
ReliabiliKtlyi:miscrhankin=g2.Thisstudymeetsthecritefroiraquality testing.However, thestudylacksinformatioonn purityofthetestsubstance and actualmeasurements ofthe amount oftestsubstanceinsolution.
REFERENCES
Test was conducted by T.R.WilburyLaboratoriesI,nc.o,f Marblehead, Massachusettsatthe requestofthe3M Company, Lab Request number P1624,1996.
OTHER
Last changed: 5/25/00-
Study Title
Acute ToXicity of rc-10!5 to the Daphnid, Daphnia magna
Data Roquirementi Guidelin* Wu=Lber U.S. EPA-TSCA, Gu4.deline 797.1300
Authors
Timothy J. Ward lacqualine Novius
Robert L. Boari
Study Completed
Kay 15, 1996
Sponsor
3H Company Env;-ro.-mental Laboratc--y
935 Bush Avenue Building 2-3E-09 St. Paul, K Innesota 55144
Testing Facility
-L-K..
ilbury 1a,l>oratories, iic. 40 Doaks Lane
head, ma saachusetts 0194 5
Lbury,gtu y N
30-TH
Page 1 0
1. TABLR Or CIOUTZINTS
StCTIOti:
I. TaLble of contents
ii. .11.
Index Of TaLbles and rigures Summary
IV. General Information
V. V-..
Introftction methods and materials
Vil. Results
Vl:l. References
IX. Signature Page
Appendix A. water Quality Data from Toxicl-',y Test
P&GE
2 3 4 5 6 6 11 15 16
17
R.@wilbury;st
Niizbe
2
11. INDZI OF TABLXB AXD FIGURM
Table I
chemical characterization of a representative sample of dilution water used for the toxicity test with daphnids, Daphnis magna, and rc-1015.
Table 2.
Test system eu=ar-y for the toxicity test with
FC-1015 and daphnids,
Z)aphnim magna.
Table 3.
survival and sublethal effect data from the toxicity test w'-th daphrids, Dephnia magns, and FC-1015.
Table 4.
median lethal concentrations (LC509) and med4-an effective concentrations (EC50s) from the toxicity test with daphn@-ds,Dap,@n.@maagna, and FC-1015.
Table A.l.
Dissolved oxygen concentration, conductivity, pH, and temperature measured during the toxicity test with daphnids, Daphnia magna, and rC-1015.
?igure 1.
Sur-vival and i=obilization of daphrids, Daphnia magna, exposed to FC-1015 for 48 hours.
PAGN 7 9
12 14
13
T It.'w
--'stud NUMD*R 1030-TH
:3
111. BLIOCILRY
The acute toxicity of rC-1015 to the daphnid, Daphnis magn&, is
described in this final report. The test was conducted for 3H Company for
48 hours from April 16 to 18, 1996, at T.R. wilbury Laboratories, Inc. in
Marblehead, Massachusetts.
It was conducted by Jacqueline Novium, Jeanne
Hagazu, Kovo,-i Stevens, Peter Kowalski, Robert Boori, and Timothy Wa-rd
according to the procedures of the U.S. Envircrmental- Protection Agency
(1993). FC-1015 was supplied by the sponsor.
The test was performed under static conditions with five concentrations of test substance and a dilution water control at a temperature of 20.2 to 21.0*C. -,hedilution water was filtered deionized
water co',Iected at Marblehead, Massachusetts aid ad)ua*,edto a hardness of 176 z@g/L as CACO,.
Nr-minal@concentrations of FC-1015 were 0 (control@, 430, 730, 1,200, 2,000, and 3,330 mg/L. Daphnids used in t;iatest were less than 24 hours old at the start of the test. They were prc>duced at T.R. Wilbur-y Laboratories and acclimated under test conditions for their entire life (cultures that produced test specimens were maintained at test conditions for more than 7 days). After 48 hours of exposure *@@iecoitrol daphnids had an average wet weight (blotted) of 0.35 Mg. All Oai)hridj3were in go<>d con,ditionat the beginning of the study.
Exposure of daphrids to FC-1015 resulted in a 48 hour median lethal concentration (LC50) of 1,550 mg/L (95% confider-,ceinterval - 1,200 to 2,000 :ng/--)and a 48 hour median effec-,;-veccncer,-ration (Ec50) of 1,200 mg/L 195% confidence interval - 730 to 2,000 mg/L). The 48 hour no observed affect concentration (NOEC) is 730 lr3/L-
ic
7 11
1030-TH
a %@,or ly
IV. GERERKL IN710RXJLTIOK
Kater@al Tested; Sponsort
Test Substance shipped Fromi
Reported Purity:
Experimental Start Datei
Experimental Date;
co-npletior.
FC-1015
3H Company Environment-al Laboratory 935 Bush Avenue Building 2-3r-09 St. Paul, Kinnesota 55144
3H Company Environmental Laboratory 935 Bush Avenue Building 2-3E-O9 St. Paul, Xinnonota 55106
10 0 it
April 16, 1996
April 18, 1996
A-rchive L@ocation for the Biological Raw Data and a copy of the Final Report.
T.R. wilbury Laboratories, Karblehead, nassachusetts
Inc.
Wilbury Study Number-,,10 0-TH,
V.
XICTITOOLICTION
This study was sponsored by 3H C=pany, St Paul, Kinnonota. The objective of the study was to determine the 24 and 48 hour LC50s and EC50a of the test substance to the daphnid, Dephnia msgna, a freshwater
invertebrate, under static conditions. The study was conducted following U.S. EPA (TSCA) guidelines but at the Sponsor's request it was not conducted according to Good Laboratory Practice (GLP) r-ules. The report contains sections that describe the methods a-ndmaterials employed in the study, and results of the investigation. The report also contains an appendix that presents water quality data collected during the test.
Vi. XETRODS AND XXTERIALS
TEST SUISSTA.NCE:
The sample of FC-1015 (T.R. Wilbur-y Lal-oratories Sample Number 604) was delivered to T.R. Wilbur-y Laboratories on April 4, 1996. The sample was delivered in a metal can within a cardboard box at ambient temperature. -he test substance was contained in I 250-ml plastic bottle. The label attached to the bottle contained the following informat@ont -rc1015, (:.R P.624), See MSDS, Susan A. Beach, 2-3E-09, (612) 778-7452, 4/2,'48-.
FC-1015 (a clear liquid) was supplied by 3M Coirpany, St Paul M,-n,iesora. Prior to use the test substance was stared in ,he dark at r;@@ toz,.perature, The test substance was assumed to have a purity of 100% active ingredient and to be stable under storage and testing conditions.'unused test substance is returned to the sponsor.
DILUTION WATERI
water used for acclimation of test organisms and for all toxicity
testing was deionized water collected at T.R. wilbur-y L&t>oratories in
K&rblehead, KassachuBetts.
water was adjusted to a hardness of
180 mg/L as
and stored in 500-gallon polyethylene ta,-.kswhere it was
CaC03
aerated and continuously passed through a particle filter, ultraviolet
sterilizer, and activated carbon. Water used for the test had a hardness'..
of 176 mg/L and an alkalinity of 108 mg/L as caco,, and it' contained..
<0.01 mg/L particulate matter and <1 mg/L total organic cark>on.',@R@esults.
of ch-ical analysis of a representative sample of water &re presented'in-.,
Table 1. A sample of dilution water was sent to Lancaster Labcratori'*!,-'
Lancast*r". PA for analysis. of 'tctal organic c&rbon.@',.The'@ sampli@-:@ra
daternine@ to contain.-c I.O.Mg/L total.org&nic c&rbo
!.. Az
study Number
Table
Chemical chiLracterizatiorL of a r*pr*sontative sa=ple of the
doionized water used to formulate the fros*iwater dilution water for the toxicity test with daphnids, Daphnis magns, and rc-ici5.
Parameter
Unit of Heasuremount
Detection Liz-t
measured value
metals XIU=Linum )Lrsenic aoron cadmium
chromium Cobalt Copper Iron
Lead mercury Nickel Sil-jer zinc
Nitrate chloride Fluoride Total Organic Carbon Total Phosphorus Organc>chlcr4.ne
Pesticides Toxaphene
Organophosphorus Pesticides
Dimethoate
TEPP Morocrotophom PCBs
mg/L mg/L mg/L Mg/L
mg/L mg/L mg/L mg/L
mg/L mg/L mg/L
mg/L mg/L mg/L mg/L
Mg/L
as N
mg/L mg/L
iig ;jg/L
;jg/L ;jg/L ;jg/L
;jg/L g/L
0.1 0.01 0.5 0.0002 D.01 0.03 O.C@5 0.03 0 .rl05
0.0003
0.03
O.Cl,
c.02 a.cs I
0.1
1 0.03
0.5 2
0.5 2.0 2.0 2.0 0.5
ml)2
ND
ND ND ND ND 0.03 ND
ND
ND
ND
ND
ND
ND
ND ND
ND ND
ND ND
ND biD
Notes:
1. Parameters were measured in doicnized water (-@-.setdo form=ulat
dilution water) that-was collected during August,@'.19955, analyzed by Paca.",'@Inc-@'as par-t of routine water @quilit testing
2. ND noi'd'at'acto at or gtl>v6e the detection 1ijait
R., w
Study N
34
79"
.Tuvenildeaphnids,Daphnismagna,employedam test organismswere less than 24 hoursold at the startof the test. They were froma single source and were identified using an appropriate taxonomic key. Test specimens were produced by adult daphnids that were maintain*d under test conditions at the T.R. wilbury facility for more than, 7 days. The original culture was obtained on Karch 26, 1996 fr= Aquatic BioSystmo, For-t Collins, Colorado. KoaaLLromentsmade during the 7 days before the start of the test indicated that the culture temperature ranged from 20.1 to 20.5*C and the dissolved oxygen concentration was always at least 6.9 mg[L. The culture produced young prior to day 12 and a submample of adults prc>duced at least 3 younq/daphnid/day for 7 days before the start of the test.
During acclimation daphnide were not treated for disease, they were free of apparent dionse, in@.ur4.es,and abno-m&litiob at the beginning of the test and there was no mcrt&14.ty during the 48 hours preceding the start of the test. The culture was supplied with a yeast/trout chow mix (Lot# I>CC2) and the freshwater alga, solonastr-um capricornutum, daily during acclimation. Daphnids were not fed during the test. Control daphni.da, blotted and weighed at the corclusicn of the toxicity test, averag*d 0.34 mg and the loading rate during the definitive toxicity test was approximately 14 mg/L.
TOXICIT"Y TESTING:
The toxicity test was perfor--ed according to the procedures of the U.S. Environmental Protection Agency @1993). A si==r-y of the test system in presented in Table 2.
A screening test was not conducted because nominal concentrations for the definitive test were provided by the sponsor. The definitive test was performed from April 16 to 18, 1996 with five concentrations of test substance and a clilution water control. Nom-inal concentrations of P'C-1015 were 430, 730, 1,200, 2,000, and 3,330 mg/L. Appropriate,. amo,.lnto'of test substan,ce were add-ad directly to d;-Iution water in test. vessels, without the use of a solvent.
Twenty daphnids were indiscriminately and equally distributed,among'.
two repLIcatsn_of each,treatment. The test was performed i@i'300 zml'glann
beakers that;. contained' 250 ml of test solution (wat
op
as
approximately 9 cm:i Test vessels were randomly arranged in
during the 48 hour -ts8t "(a random numj>ers tat)le was used"@@64i
he
i 7@e-location of.oach v,essol) and they were loosely c ver,d
The t.04@-'w'.1 cp.!'@ft".d riod @witt
.i.d rlsity of,130,footcandles"
@6' our,'@light.@ancliS,@hoUr@dar
e transition
ri'
cai
fluorescent @lights",4t a
@T. R. Wilbur3 @'studli4
1030- TH
-page.. OZ i'19
Table 2.
Test system summary for the toxicity test with TC-1015 and daphnids, Daphnla jugns.
Sp4bCi@t
I
)Lge/Size of Test organismst
Test Du--ationt Test Vessels:
Depth of Test Media:
Namber of Replicate Test Vesselst Organia=S per Replicates
Temperature Ranges
M;-ni-m,= Dissolved oxygen:
Phctcpericd:
Light Intensity: water Quality measurements:
End Points:" Acceptability Criteri
Daphild Magna
Juveniles/-0.35 mg, wet weight
48 hours
300 ml glass beakers
9 cm
2
10
20.2-2i.Olc
7.9 mg/1-
16 hours light and 8 hours dark with a 15 ir.@ni;tteransition period.
130 footcandies
Dissolved ox-ygen, pH, temperature, and conductivity daily in each test vessel. Ha--dness and alkalinity in dilution water at the star-t of the test. Temperature measured at least every hour in t reprocantative test vessel. Dilution water analysis is suz=rized in Table 1.
24 and 4B hour LC50 and EC50; 48 hour NOE^,.
510% control mortality and acceptable temperat,.ire range.@
7 t @i
The number of surviving organisms and the occurrence of sublethal effects (i=obilization, loom of equilibrium, loss of reflex, excitability, discoloration, or change in behavior) were determined visually and recorded initially and after 24 &ad 48 hours. Dead test organisms were removed when first observed. Dissolved oxygen (Ysl Kodal 57 mt*r; instrument nu=er 1), pH (Beckman model pHl 12 motor; instrument number 144), conductivity (Cole Pa.-=r motor, instrument mL=Ll>er94). and
temperature (Backman model pHI 12 meterl instrument number 144, probe pHT5) were measured and rocord*d daily in each test chamber that contained
live &ni=alx. The temperature in on* test vessel was recorded at least hourly during the test.
STJLTISTIrAt KETHODSI
Results of the toxicity statistic&! tectiniques. Computer
test were imterprotod methods IStephan, 1983)
by standard were used to
calculate the 24 and 48 hour LC509 based on mortality and the EC50a based
on i=obilization.
All calculations eere performed using nominal
concentrations of test substance. The no observed effect concentration is
the highest concentration of test substance .hat did not cause any
tox'-cant-related mortality or sublethal effects.
2. wilbury study Number DjO,.
p
Vil. RLSULTS
Insoluble material was not noted during the test. Biological and
water quality data generated by the acute toxicity test are presented in Table 3 &ad kppendix A, resp*ctively. The measured concentrations of
dissolved oxygen ranged from 7.9 to 9.2 mg/L, the temperature ranged from
c
20.2 to 21.7'c, conductivity ranged from 610 to 800 ;nmhoo/cm, and the pg
ranged frcim 8.0 to 8.3. on* hunclred percent survival occurred in the
control exposure and these daphnids did not exhibit any subletha-Ieffects.
The 24 and 48 hour LC50s and EC50s for daphnida exposed to rc-1015 are presented in Table 4. The 48 hour LCSO for daphnids exposed to rc1015 in 1,550 mg@L, with a 95% confidence inter-valof 1,200 to 2,000 mg/L and the 48 hour EC50 is 1,200 mg/L, with a 95% confidence int r-valof 730
to 2,000 mg/L. Th" 8 hour NOEC is 730 mg/L.
00 H
T&bl* 3.
survival and sublethal effect data from the toxicity test with daphnids, Daphnia magna, and TC-1015.
Nominal
Concentration of rc (mg/L)
Rep.
Number A-live
0
24
48
hr
hr
hr
Number Affected
0
24
48
hr
hr
hii
0 @control)
1
10
10
10
2
10
10
10
430
730
-
1
10
10
10
2
10
10
10
1
10
10
10
2
10
10
10
1,200
1
10
10
10
2
10
- 10
9
2,000
1
10
10
0
2
10
10
1
3, 330
1
10
2
0
2
10
0
0
0
0
0
0
:o
0
0
0
0
0
0
0
0
0
0
0
0
0
2
3
0
1
6
0
7
0
4
0
2
0
Notat Affected daphnids were
bilized.
7-,
T.Rl.@,.WilburyStudy N
0;..T
20
is 14 12
8
4 4-
0 430
-------
730
1200
2000
3330
Concentrationof FC-1015
live Daphr@ds
Mobde Dap ds@
Survival and i=obilization'ofdaphnids, Da -exposed to rc-1015 for 48 hoLLrs
Table 4.
Kedian lethal concentrations Daphnis magna,
concentrations (LCSOO) and median (EC50s) from the toxicity test with and FC-1015.
effective daphnids,
Exposure paric>d
LC50/zcso
95 percent confidence limit
calculation me tbod
tzso
24 hours
2,730 mg/L
2,000 to 3,330 mg/L
Binomial
48 hours
1,550 mg/L
1,200 to 2,000 mg/L
Binomial
rcso
24 hours
---1,200 mg/L
730 to 2,DOO mg/L
BinouLial
48 hours
1,200 Mg/L
730 to 2,030 Mg/L
Binomial
VA
c@ c,
Li
-7
70
LAO,
EP@ PROBIT ANALY4;IS PROGRi4M USED FOR CALCULA T@N(i EC VAL@IFS
Version 1.4
. 16 24 24 -lirs exdi
Coric.
Nunioe,
Di &3tknt a m-3orva.
i@u m
ber Rctsip.
Obsier vied
-Pr-.iIF3c3r
,-')Ot;
2", 21:1
sa[3 5/ 22,*96
Adju"stio4d@,
@Propiair 'Rp%ponding,
vies pcyn
t,"t
-?itv
in'?t F intf@-r@cept Tb
t k,1:" L.
.4.4
2
7. 1 i-i:)47, l.L61--i
'-Siidnta@i9PLtS
46'
1624 .24-hrs exp.,,, Dapt@ni,a magna mg /Ll
4ab 5/ 22/96
Eatima ted EC Values apid Confideride;Litilts@-,-.
Conc.
Lower:.
lipper,
95% c o'nfidenC*!-LLm3. s;
-.00 EC50.
844 40 60'
I t'91a 12.'3,j.6676. 17F39. L@6r,14
88 1 .4062 @,991
-0 04'?
EC,,,'-j.
z,
2ZiZ6.S74!3
6,41
-7.9
?ta'
F,,-l r)n 1,3' m -1,9n 3
L
PLOT OF ADJLjSl@-1) F-RI)PITS AN!) PREI)ICTED
LINE
viii. RZTZRM=s
AsTm. 1986. standard Practice for conducting Acute -oxicity Tests with Fishes , Kacroinvortabrates, and Amphibians. E-729-80 - In Annual Book of standards.
1
,-
Stephan, C.E. 1983. computer.progr am
U.S. EPA.-Duluth, mN. personal
for the Calculation cc=munication.
of LC50 values.
U.S. EPA. 1993. Toxic substances Rules. section 797.1300.
Control Act Test Gzidelines;
Final
tu
r
IX. BlONATuitz PACC
Jac4uelin* M. N*viUs StQdy Director Biologist
Robert L. Boori Coauthor
Jeanne Aquatic
P. Mag@zb Toxic-o@ogist
For-. 'Kaven Biologii3t
M. Stevens
Pete@'L. Kowalski
by@4j'.@,liard
,Y
Wil
$tUd Numb-
I)vd
oq=,14,. pn;g
qll
Z$2.L IZI:)ixox "cmi vxyci iii7vnO
xTPuDddy
Table A.I.
Dissolved oxy(jon concentration, conductivity, PH, and temperature measured during the toxicity toot with daphaids
Daphn.LAmagna, and ]rC-1015.@'
No3tinal Concentratioa
of rc-1015 (mg/L)
Rep.
Dissolved oxygen (mg/L)
0
24
48
hr - hr
hr
conductivity (gmhomlcm)
0
24
48
hr
hr
hr
0 (control)
1
2
430
1
2
730
1
2
1,200
1
2
2,000
1
2
3,330
1
2
9.1
7.9
9.1
8.1
9.1
8.2
9.1
8.0
9.1
8.1
9.1
8.5
9.1
8.3
9.1
B.1
9. 1 8.1 9.1 F 8.3
9.2
8.6
9.2
8.5
8.2 8.3
8.3 8.3
8.3 8.3
8.4 8.3
8.2 8.2
8.3 --
61@o
610
620
-7@
610
610
620
630
630
630
630
630
640
650
650
650
650
650
660
680
670
680
660
680
680
720
710
720
720
710
710
800
780
780
800 790 7t
R v lbuz-y 0-.u
r
,,57
T&ble A. I. Continu*d.
N=inal Concontration-,
of IPC-1015 (mg/L)
Rep.-"
0 (control)
1
2
430
1
2
730
1
2
1,200
1
2
2,000
1
2
3,330
1
2
Tomperature i-C)
0
24
48
hr
hr
hr
20.4 20.4
20.3 20.4
20.3 20.2
20.2 20.2
20.3 20.3
20.4 20.4
20.7 20.7
20.6 20.6
20.4 20.3
20.6 20.7
20.6 20.8
20.4 20.7
20.6 20.5
20.B 20.9
21.0 21.0
20.7 20.5
20.6 20.6
20.8 --
PH
0
24
48
hr
fir
hr
8.3
6.3
8.2
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.3
8.2
8.2
8.3
8.2
8.2
8.3
B.1
8.2
8.2
6.1
8.2
8.2
8.0
B.2
82
8.0
8.2
T R','WilburY Study7Numb-- 1030-Tli . . .. .. .