Document VGQmr4Y75mgDqOB7LX903gz1o
AR226-2798
DuPont BLR 61-94
Study Title
STATIC, ACUTE, -o-HOUR LC50 OF H-19704 TO BLUEGILL SUKFISH, Lepoils acrochirus
Laboratory Project ID "askell Laboratory Report He. 61-94
Data Requirement
United States Environmental Protection Agency
40 CFP 797.1400 Fish Acuf Toxicity Test
Toxic Substances Control Act
Author Keith E. Pierson, Ph.D.
Study Completed On 26 August 1994
Perforning Laboratory
Haskell
.. I. du Font Ie Ne>ours and Company
Laboratory for Toxicology and Industrial Elkton Road. Post Office Box 50
Newark. Delaware 19714
Medicine
^Medical Research Project &--
Page 1 of 20
^^"^"^"...con.a.nTsCACB,
GOOD LABORA-m PRACTICE COMPLIANCE STATEMENT
This study was conducted in compliance vith EPA PIFRA (40 CFR 160) and EPA TSCA (40 CFR 792) Good Laboratory Practice Standards, OBCD Principles of Good Laboratory Practice (C(81)30(Pinal), Annex 2), and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) except for the deviations documented
and explained belov.
1. Stability and solubility of the test substance under test and storage
conditions vere not determined.
2. Tetranin flake food was not tested for pesticide content.
The deviation(s) did not affect the integrity of the study.
Submit ter:
Sponsor:
E. I. du Font de Nenour..: and Company
DuPont Elastomers
t. I. du Pont de Nemours and Company
Study Director:
Ai/^C 4. P/'CMf^
Keith B. Pierson, Ph.D. Research Yoxicologist
CoBpany Representative: Registration Manager
Date
^ ffu^osrHf^
^'S^^.--^^.^
DuPont Bill 61-94
TABU OF LUW1BM1S
Page
Good Laboratory Practice Compliance Statement
2
Table of Contents
General Information
.............. Su--ary
............. Quality Assurance Docuaentation
.......................... I. ................. II. ................. III.
Purpose Records
Materials and Methods A. Test Substance
.......... B. Dilution Water . . .
............ C. Aniaal Culture . . .
D. Test Methods .
.
.
B. Chemical Analysis .
3 4 .
5 6 .
7 7 7 7 7 7 7 8
P. Statistical Methods
8
IV. Results and Discussion
8
V. Conclusion
9
.................... VI. Literature Cited
9
......................... VII. Tables
10
....................... Table 1. Chemical Characteristics of Baskell Laboratory Veil
............................ Water
10
....................... ymSllWI^ Table 2. Water Chemistry of the Dilution Water at Test Start 11
Table 3. Physical^ard Cheaicaj. Characteristics of
B-1970&
Test Solutions .
12
Table 4. Mortality ot BluegiHSunfish, Lepoais
acrochirus, Exposed to B-19704TWU
96 Hours in a Static, Unaerated TeSt
14
Table 5. Behavior of BluefiU, Sunftsh, Lcpoais acrochirus,
Exposed to H-19704j4BIMBBfjll!oF*96 Hours
in a Staj.ic, Unaeragg Test
16
Table 6. H-19704lfBB--Bdconcentrations Causing
. . . . . . . . Mortality-to 50Z (LC50) of Bluegill Sunfish,
Lcpoais acrochirus
17
Table 7. Standard Lengths and Wet Weights of Control Fish
.............. At Study End
18
VIII. Appendices
19
.................................................. 1. Protocol Aaendaents and SQP Deviations
19
Company Sanitized. Does not contain TSCA CBl
Material Tested;
Haskell Huaber; Purity;
Composition; Contaainants:
CAS Registry Nmber;
Sponsor:
Study Initiated/CoBpleted; In-Life Study
Initiated/CoBpleted;
GHBAL IHFOUtATION Octanolc acid, pentadecafluoro-, saa-aniw
salt
H-1970A
S-^
None reported 3825-26-1 DuPont ElastoBers
E. I. du Font de Leiours and Competv
Viloington, Delaware 19898
13 August 1993 / 26 August 1994
23 August 1993 / 27 August 1993
Company Sanitized. Does no? contain TSCA CB;
DuPont BLR 61-94
suMton
H-19704 exhibited low toxicity to bluegill sunfish, Lepoais nacrochirus, in a 96-hour, unaerated, static test vith noinal concentrations of 262, 328, 410,
512, 640, 800 and 1000 Bg/L. The 96-hour LC50 based on nominal concentrations
was 634 Mg/L with a 95Z fiducial interval of 567 to 725 ig/L.
Work by: tA^y.%^A M.
Setfrey T. Turner, Sr. Toxicology Associate
t /-' <, A
Ho/Day/Year
^^-^UGhM.je-
Laurie D. Bouchelle Toxicology Technician
^ / ^ ^ Mo/Day/Year
THIS REPORT IS APPROVED FOR ISSUE.
/^ ft'^J &.
Keith B. Pierson, Ph.D. Research Toxicologist and Study Director EnvironBental Sciences
AutrViT ^ W^
Ho/Day/Yeat
Company Sanitized. Does not contain TSCA CBl
Dates of Inspection:
Conduct - 6 23/93
Records, Re:.: rt(s) - 8/9-10/94
Date(s) Findings Reported to; Study Direc::r - 8/10/94 Management - 3/24/94
Reported by:\^_^-_KA_^ J<_ H-^-ll
Donna R. Holt Quality Assurance Auditor
Ool^
Dat
Coinaany.SanitizecL Dcss noi contain TSCA C3:
i. nuosx
The goal of this study was to assess the acute toxic!ty of H-19704 to Juvenile blueglll sunfish, Lepoais acrochirus.
II. RECORDS
All data and 'ie final report vill be aalntained either at Haskell Laboratory
for Toxicology and Industrial Medicine (Newark, DE) or be archived at the DuPont Records Manageaent Center (Vilaington, OB).
A. Test Substance
III. HATERIALS AM) METHODS
The teat substance is described under General Intonation. Test solutions were
prepared by weighing out the appropriate quantity of test substance, adding it
to 10 liters of dilution water (see below) and stirring. The test substance appeared to be entirely in solution at study start. Stability and solubility
of H-19704 in dilution water were not deterained.
B. Dilution water
Water (Table 1) originated froa the Haskell Laboratory well.
C. Anl--1 Culture
Bluegill sunfish, Lepoais acrochirus, were purchased froi either Northeastern
Biologists (Rhineback, NT) or Aquatic Research Organises (Haapton, NB). Fish were acclimated in continuously-flowing, well water in 545-L circular, fiberglass tanks for 212 days at a aean teaperature of 21.2C (rangefinder) or for 32 days at a aean teaperature of 22.6'C (definitive study) prior to study
start. Pish were fed newly-hatched, live Arteaia sp. (Aquarliu Products, Glen
Burnie, ND), frozen adult Artcaia sp. (Golden Gate Brand, Kordon Division,
Novaiek, Inc., Hayward, CA) and/or Tetraain* flake food (TetraUerke, Geraany) generally twice daily on weekdays and daily on weekends. Fish were treated with 2-3 parts per thousand salt (NaCI) on days 0-4 of the 212-day acclimation (rangefir.der). Pish were treated with three handfuls of salt (NaCI) on day 0, '200 parts per Billion foraalin for one hour on days 1-3 of the 32-day accliaation (definitive). Sickness, injury and abnoroality were not present at study start. Aniaals were identified by labels on aquaria and test vessels.
D. Test Methods |1]
Seven test concentrations and a control were used. were 262, 328, 410. 512, 640, 800 and 1000 Bg/L.
Noiinal test concentrations
Glass aquaria [20 liter; 39.9 (length) x 19.8 (width) x 25.5 ci (height)j
Company Sanitized. Does not contain TSCA CBJ
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OuPont BLR 61-94
f t containing 10 L of
solution (approximately 13.5-ca liquid depth) were
employed. Positions of test chambers in the water bath used to aatntain constant teaperatu.-A vere assigned uaing random numbers. Ten fish vere added
to each replicate using randoa numbers (two replicates per concentration; total 20 fish per concentration). Control fish ranged froa 1.2 to 2.8 ca standard length (Man 2.1 ca) and 0.0451 to 0.524 g vet weight (Bean 0.228 g). Control loading vas 0.23 g/L at test conclusion. Test solutions ranged from 21.3 to 22.I'C (mean 21.7C). Pish vere not fed approxiaately 24 hours prior to nor during th'j test. A photoperiod of 16 hours light (312 to 344 Lux) versus 8 hours darkness was eaployed vlth 25 minutes of transitional light (<2.15 Lux) preceding and following the 16 hour light interval. Mortality and behavioral
effects were noted daily.
Dissolved oxygen (TSI Model 54A, Yellow Springs Instruaents, Yellow Springs, OB), pH (Corning Model 610A, Corning Scientific Instruaents, Medfield, MA) and
temperature (Arerican Society of Testing Materials registered aercury theraoaeter) were aeasured in each replicate before addition of fish at test
start, dally, at total fish aortallty and/or at test end. Total alkalinity
(Bach Cheaical Coapany, Loveland, CO), EDTA hardness (Bach Chealcal Corpany, Loveland, CO), and conductivity (YSI Model 33, Yellow Springs Instruments,
Yellow Springs, OB) of the control water were measured before fish were added
at test start. A continuously-recording thermoaeter in a water-control replicate was used to check temperature variations during the 96-hour test.
B. Chemical Analysis
Cheaical analyses of the test substance concentrations in water were not perforaed because a suitable method was not available.
V. Statistical Methods The 24-, 48-, 72- and 96-hour LC50s and their 95% fiducial intervals were calculated using probit analysis (2].
IV. RESULTS AfO DISCUSSION Protocol aaendments and/or SOP deviations appear in Appendix 1.
A rangefinder was conducted. Nominal test concentrations were 0, 0.1, 1, 10, 100 and 1000 g/L. Mortality after 96 hours was 0, 0, 0, 0, 0 and 80Z, respectively. The definitive study was conducted using noainal B-19704 concentrations of 262, 328, 410, 512, 640, 800 and 1000 g/L. We planned to aeasure B-19704 concentrations in test solutions, but the suggested analytical method failed.
Preliminary research suggested it was not cost effective to develop a new
method. Bence, nominal B-19704 concentrations are reported herein.
Con^nv Sanifc". ^ Rsi conia!?l TSCA a"
DuPont BLB 41-94
All eh--leal and physical parameters were within expected rang-- (Tables 2-3).
f/L The 96-hour LC50 (Tables 4 and 6) was 634
vith a 95X fiducial interral of
567 to 725 ag/L based upon noalnal concentrations. Behavioral observations are
reported in Table 5. Fish Retries are reported in Table 7.
v. concufsioN H-19704 exhibited low toxicity to Juvenile bluegill sunfish, Lepoais aacrochirus, in an unaerated, static test. The 96-hour LC50 was 634 ag/L (95X fiducial interval of 567 to 725 g/L) based upon noaiinal test concentranons.
1.
test.
2. 333 p.
40 CFR 797.1400.
VI. LITBATuRB CITBD Toxic Substances Control Act.
Fish acute toxicity
Finney, D. J. 1971. Probit Analysis. 3rd Ed., Cambridge Uni-.. Press.
' C.mp.S,-ze<.D.esn..c.n."TSCACB
BOD, g/L COD, g/L
DOC, g/L TOC2, g/L Total Kjeldahl N, mg/L A--onia N, g/L Turbidity, NTU Phenolics, g/L Color, apparent Co/Pt3
Solids, total suspended, mg/L
AluninuB, g/L
Antimony, ag/L
Arsenic, g/L Berylliic, g/L
Boron, mg/L
Brfide, g/L
CadBiiw, g/L
Calclua, g/L Chloride, g/L
ChroBiuB, g/L Cobalt, g/L Co"per, g/L Cy nide. g/L
Iron, g/L Fluorlde, g/L
<7
<0.2 <0.2 <0.01
<O.G1
<0.04 <0.5 <0.01 24.5 6.9 <0.05 <0.05 <0.02 <0.005 <0.1 0.1
Lead, g/L HagnesluB, ag/L
MBAS/LAS, Bg/L
Mercury, ag/L Nickel, g/L
Nitrite, og/L
Nitrate, ag/L
Ortho-Phosphate,
Potassium, Eg/L Selenium, og/L
Silver, mg/L
Sodlua, Bg/L
Sulfate, ng/L Sulfide, ng/L
Zinc, ng/L
Bg/L
<0.1 3.8
<0.02 <0.00020
<0.05 <0.1
0.5
<1
1.8 <0.005 <0.02
8.7 5.2 <0.10 <0.04
Volatile priority
pollutants
ND4
Acid extractable
priority pollutants ND5
Base/neutral
priority pollutants NO5
Pesticides/PCBs
W
Organophosphate
pesticides, vg/L
ND*
lD&te of s--ple collection 12 May 1993 unless Indicated otherwise, analyses performed at Lancaster Laboratories, Lancaster, Pennsylvania; 'Date of sample collection 23 March 1993, analyses perforoed at Lancaster Laboratories,
Lancaster, Pennsylvania; 'Units based on cobalt/platinun reference; None detected; level of detection ranged froa < 5 < 100 vg/L; 'None detected; levels of detection ranged fron < 10 to < 25 ug/L; "None detected; levels of detection ranged froa < 10 to < 100 ug/L; iflom detected; levels of detection ranged froa < 0.01 to 4 ug/L; None detected; levels of detection tanged froo < 0.01 to <
0.1 ug/L.
10
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DuPont HLR 61-9*
Static, Acute, 96-Hour LC50 of H-19704 To Blueglll Sunfish, Lcpoaia --crochirua
TABLE 2
Vater Chtnistry of the- Dilution Hater at Test Start
Total Alkalinity (ng/L as CaCO))
79
EDTA Hardness (ag/L as CaCO;)
76
Conductivity (unhos/cm)
165
11 Company Sanifasd. Does not contain TSCA CBl
-.^Sifca-f;
TABIf 3 Physical and Cheaical Characteristics of 3-19704
Test Solutions
Nominal
Concentration (ng/L)
0
A
B
24
A
B
Hours
48
A
I >
72
A
I 1
96
1 L
Dissolved Oxygen1' (Bg/L)
- Control
8.5
8.5 7.8
7.9 7.5
7.,6
7.3
7. 5
7.,1
2d2
8.5
H.5
7.U
/.(!
7.(,
7. B
7.A
7. 5
7,,0
328
8.5
8.5 8.0
8.1 7.6
7.,6
7.3
7,.3
7,.0
410
8.5
8.5 7.9
/.9 7.5
7,,7
7.1
7..0
7,,1
512
8.5
8.5 8.0
8.0 7.7
7 ,7
7.3
7..2
7,.0
640
8.5
8.5 7.9
7.8 7.5
7,.6
7.0
7,.2
6..9
M
800
8.5
8.5 8.1
8.1 7.6
7,.5
6.9
7,,0
6 ,7
1000
8.5
8.5 8.2
8.1 7.7
7,.6
---
7,,0
--
pB
Control 262 328 410
512 640 800
1000
7.4
7.4 7.4
7.4 7.,
7.3
7.4 7.4
7.4 7.4
7.4
7.4 7.4
7.4 7.4
7.4
7.4 7.5
7.4 7.4
7.2
7.1 7.3
7.4 7.3
6.9
6.9 7.3
7.2 7.2
6.9
6.9 7.3
7.3 7.2
7.0
6.9 7.2
7.2 7-2
7 .4 7.3
7 .4
7 .4
7 .3
7.4
7 .3
7 .4
7 .4 7.3
7 .3
7 .3
7 .4 7.3
7 .4
7 .3
7 .4
7.3
7 .4
7 .3
7 .2 7.2
7 .2
7 .2
7 .3 7.3
7 .3
7 .3
7 .2
---
7 .1
''
All fish had previously died so no reading was aade. theoretical saturation at 22C was 8.8 mg/L.
Company Sanitized. Does not co
t
TABLB 3 CONTINUED
Physical and Chemical Characteristics of B-19704J
Test Solution
Noiinal Concentration
(g/L)
Temperature (C)
Control
262 328 410 512 640 800 1000
0
A
B
24
A
Hours 48
BA B
72
96
A BA
22.1
22.1 22.0 22.1 22.1 22.1 22.1 22.0
22.1
22.0 22.1 22.1
22.0 22.1 22.0 22.0
21.6 21.6 21.6 21.7 21.7 21.6 21.6 21.6
21.7
21.6 21.6 21.6 21.7 21.6 21.6 21.6
21.8 21.7 21.7 21.7
21.7 21.7 21.7 21.8
21.9
21.7 21.7 21.7 21.8 21.7 21.8 21.8
21.6 21.5 21.4 21.5 21.6 21.5 21.5
.I--*
21.5 21.5 21.4 21.4 21.6
21.6 21.5 21.6
21.5 21.4 21.3 21.4 21.5 21.4 21.4
--------
All fish had previously died so no reading was made.
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DuPont HLB 1-94
f
Static, Acute, 96-Hour LCSO of H-1^704 To
1
Bluegill Sunfish, Lepoals nacrochims
TABL2 4
W------f Mortalityr of Bluegill Sunfish. IfWatis acrochims,
Expos ed to H-19704
Jror 96 Hours
in a Static, unaeratedTest
Mortality In Percent
Noainal
Concentrations
(t/l.)
24
Water Control
A*
0
B*
0
Total
0
|w>
262
A
0
B
10
Total
5
328
A
10
B
10
Total
10
410
A
10
B
20
Total
15
Tioe (Hours)
48
72
96
0
0
0
0
0
0
0
0
0
0
0
0
10
10
10
5
5
5
10
10
10
10
10
10
10
10
10
10
10
10
20
20
20
15
15
15
A and B represent replicates; each replicate contained 10 fish (total 2 : fish per test concentration) at study start.
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DuPont HUt Sl-94
Static, Acute, 96-Hour LC50 of H-197C34 To Blueglll Sunfisb, LepoBis acroehlrus
TABLE 4 aMtUIHW
Mortality of Bl uegillSunfish, Lenglis iacrochirus,
1
Exposed to B-1970401
B9)^or 96 Hours
in a Static; UnaeratecTTest
Mortality in Percent
Noainal
Concentrations
(Bg/L)
24
Time (Hours)
A4H8
-7"2'
-
-96
512
A"
30
30
30
30
B"
30
30
30
30
&
Total
30
30
30
30
640
A
40
40
40
40
B
40
40
40
40
Total
40
40
40
40
800
A
70
70
70
70
B
60
60
60
60
Total
65
65
65
65
1000
A
90
B
90
Total
90
10090 95
100
100
90
90
95
95
A and B represent replicates; each replicate contained 10 fish (total 20 fish per test concentration) at study start.
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DuPont BLR 661^^44
Static, Acute., 99<66--Hour LC50 of B-19704 T<o9
BBlulugf(lill Soranfish, Lepoalis aaccrroocchilruas
UUM5
B4ehavlor of Bluegm junfish, Leggais acrocb irus,
Exposed t o H-1970A'
"xor 96 Hoturs
in a Scat rc'ttiaeratenrest
|
NoBinal Concentrations
(g/L)
Water Control
A B
262
A B
328
A B
410
A B
Nuiber Shoving Behavior/Total Number of Fish Alive
I
ii
24
0/10 0/10
0/10 0/9
0/9 0/9
i'
Tiae (Hours)
ii,
48
0/10 0/10
72
96
f
f,
------------------------
1:
^
0/10
0/10
*
0/10
0/10
t
0/10
0/10
0/10
i
0/9
0/9
0/9
i
t.
'1
}
1
0/9
0/9
0/9
0/9
0/9
0/9
0/9
0/9
0/9
0/9
0/8
0/8
0/8
0/8
i
AB 00//77 00//77 00//77 00//77 :I, ii
640
;?
I "" A 0/6 0nf/t6- 0/6 0/6 ''|
B . 0/6 0/6 0/6 0/6 'y
800
A
,a/3
3la-//3I
3->-.-/3
3- -/.3-
1
B
4/4
4"/4
4-/4
4"/4
1
i
1000
A B
rr//ii
----b
r/i
----b
r/i
--
r/i
Lethargic.
^
"All fib had previously died.
16
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\-
--^i
J
Tine
LC50
<Bg/L")
24 Hours
647
48 Hours
634
72 Hours
634
96 Hours
634
952 Fidui:ial Litits
(Bg/L)
(fflg/L-)
575
746
567
725
567
725
567
725
Based upon nooinal concentrations.
Slope
4.83 5.09 5.09 5.09
Y-Intercept
-13.6 -14.3 -14.3 -14.3
17
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DuPont ELB (1-94 Static, Acuf, 96-Hour LC50 of H-19704 To
Bluegill Sunflfh, Lpoal acrochirua
TABLE 7
Standard Lengths and Vet Weights of Control Pish
At Study End
Replicate
A A A A A A A A A A
B B B B B B B B B B
NiiBber
1 2 3
4
5 6 7 8 9 10
1 2 3 4 5 6 7 8 9
10
Standard Length
(ca)
2.3 2.1 2.1 1.9 2.3 2.3 1.2 2.6 1.9 2.2
2.8 2.2 2.1 2.1 1.8 1.7 1.6 2.6 2.3 2.0
Vet Weight
)
0.256 0.194 0.176 0.159 0.262 0.244 0.0451 0.415 0.183 0.224
0.524 0.221 0.226 0.202 0.142 0.100 0.103 0.456 0.269 0.153
18 TSCACBl
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mssa i
PKOTOCOL AMBBMBRTS AMD SOP DBVIATIWS PROTOCOLANBNDNBNTNUMBER ONE
a. The ratio between test concentrations is 0.8.
b. Test
iiirbers.
concentratijns should (Note: Randoa
be
assigned
to
test
vessels
using
randoc
vessels
in
water
bath
numbers are also used to assign location and to assign fish to test vessels.)
of
test
c. Take added at
saaples for cheaical analysis test start and at test
from
each
replicate
before
fish
are
analysis.
end. Saaples should be refrigerated prior to
d. Because of the snail (not 48) hours.
size of
the bluegills,
they
will
be starved
fcr 24
e. TOC, COD performed.
and
paniculate
analyses
of
the
dilution
vater
will
not
be
f. Purity and stock checks will not be performed.
f. pH and DO readings should be made on
study start, dally thereafter, at study
all
replicates
prior
to
adding
fish
at
replicate.
end and/or at total nortallty :a any
h. Dead fish in the controls do not i.^ed to be weighed and measured. 1. Test is static (not static reneval).
rje. pliMceaatseusrealtensgtuthdsy eanndd. weights of all surviving fish in both of the control
19
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DuPont HUt 61-94
Static, Acute, 96-Bour LC50 of H-19704 To Bluegill Sunfish, Lcpomis macrochirus
APPBOIX 1 (CONriNUBD) PROTOCOL AHBXMHfTS AW SOP DEVIATIONS CONTINUED
PROTOCOL AMENDMENT NUMBER TWO
Ve planned to measure H-19704 concentrations in each test solution. Samples
for analysis were taken and the analyses vere attempted. The analyses failed
and preliminary research suggested that it was rot cost effective to develop a
new Bethod. The samples vere therefore discarded and nominal concentrations reported. Since the purpose of this study was tc provide acute mortality data fov designing a bioconcentration test, this procedure was considered acceptable.
In 1994 Haskell Laboratory adopted a new systee for handling financial affairs.
As a result, it was necessary to develop a new set of test codes for aquatic
studies. Hence, the study code (226A) is incorrect. The correct new study code is 226.
PROTOCOL AMENDMENT NUMBER THREE
This study was performed under SOP AQ018-T-002 (not AQ018-T-001 as shown in the upper righthand corner of the Short-Term Study Protocol Sheet). The current name of the sponsoring business is DuPont Elastomers (the previous naire was Polyers for which Poly was the abbreviation which appears on the Short-Term Study Protocol Sheet).
SOP Deviations a. Test concentrations were not assigned to test chambers using random
numbers.
20 ^ mpany Sanitized. Does not contain TSCA CBI