Document RjddYoXe80pjNjdy020p0jgYa
AR226-2798
DuPont HLR 61-94
Study Title STATIC, ACUTE, ro-HOUR LC50 OF B-19704 TO BLUEGILL SUNFISB, Lepomis macrochirus
Laboratory Project ID "askell Laboratory Report No. 61-94
Data Requirement United States Environmental Protection Agency
40 CFP 797.1400 Fish Acut'Toxicity Test Toxic Substances Control Act
Author Keith B. Pierson, Ph.D.
Study Completed On 26 August 1994
Perforaing Laboratory
.. I. du Pont de Nemours and Company laskell Laboratory for Toxicology and Industrial Medicine
Elkton Road. Post Office Box 50
%Medical Research Project
Page 1 of 20
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DuPont BUt 1-94
GOOD LABORATnY PRACTICE COMPLIANCE STATEMENT
and explained below. 1. Stability and solubility of the test substance under test and storage conditions were not determined. 2. Tetramin flake food was not tested for pesticide content.
The deviation(s) did not affect the integrity of the study.
Submitter: E. I. du Pont de Nemours and Company
Sponsor: DuPont Elastomers I. du Pont de Nemours and Company
Study Director: Keith B. Pierson, Ph.D. Research Toxicologist
Company Representative: Registration Manager
Date
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DuPont HLR 61-94
TAUB OF UNMUTS
Page
Good Laboratory Practice Compliance Stateaent ............. 2
Table of Contents ................................... ' General Inforaation ................................. *
Sunary.......................................... jj Quality Assurance Docuaentation........................
I. Purpose....................................... 7
II. Records....................................... L
III. Materials and Methods............................. ' A. Test Substance................................ '
B. Dilution Water................................ 1
C. Aniaal Culture................
1
D. Test Methods................................. 1 B. Cheaical Analysis ............................. P. Statistical Methods........................... j?
IV. Results and Discussion............................
V. Conclusion .................................... *
VI. Literature Cited................................. '
VII. Tables........................ . * * ` * * ' * 1U Table 1. Chemical Characteristics of Haskell Laboratory Well
...................................................
Table 2. Water Chemistry of the Dilution Water at Test Start 11
Table 3. Physical nrd Cheaical Characteristics of
H-197Q4
Test Solutions.......... 12
Table 4. Mortalityoi Bluegill Sunfish, Lepomis --
aacrochlrus, Exposed to H-19704] p M B H P | for
96 Hours in a Static,UnaeratedTWt ............ 14
Table 5. Behavior of BluegiU, Sunfish, Lepomis aacrochlrus,
Exposed to H-19704
96 Hours
_
in a Static, UnaerdjgHT e s t ................... 16
Table 6. H-19704(HHB^fi3Concentratlons Causing Mortal!ijrto 50Z (LC50) of Bluegill Sunfish,
Lepomis aacrochlrus . . ..................... 17
Table 7. Standard Lengths and Wet Weights of Control Fish
-
At Study End...............................
VIII. Appendices....................................... 7* 1. Protocol Amendments and SQPDeviations................ "
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DuPont ILK 61-94
(3HE8AL IMPOUfATION
Material Tested:
Octanoic acid, pentadecafluoro-, anmoniua salt
Haskell Nuaber:
H-19704
Purity:
Coaposition:
Contaminants:
None reported
CAS Registry Nuaber:
3825-26-1
Sponsor:
DuPont Biastoners E. I. du Pont de !.enours and Company Vilaington, Delaware 19898
Study Initiated/Coaplcted:
13 August 1993 / 26 August 1994
In-Life Study Initiated/Coapleted:
23 August 1993 / 27 August 1993
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SUMMARY
B-19704 exhibited low toxicity to bluegill sunfish, Lepowis acrochirus, in a 96-hour, unaerated, static test with noainal concentrations of 262, 328, 410, 512, 640, 800 and 1000 ag/L. The 96-hour LC50 based on nominal concentiations was 634 ag/L with a 95* fiducial interval of 567 to 725 ag/L.
Work by: Jeffrey T. Turner,7 Sr. Toxicology Associate
* !*4 h i1
Ho/Day/Year
M n ijjw,) Laurie D. Bouchelle
Toxicology Technician
Ho/Day/Year
THIS REPORT IS APPROVED FOR ISSUE.
&-P)
Keith B. Pierson, Ph.D. Research Toxicologist and Study Director Environaental Sciences
AutrvtT 2 (. m i Mo/Day/Yeat
Company Sanitized.DoesnotcontainTSCA CBl
QUALITY A55URANCE DOCUMENTATION (H-19704)
Dates of Inspection: Conduct - 6 13/93 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: Donna R. Holt
Quality Assurance Auditor
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DuPont BLR 61-94
I. NRPOSI
The goal of this study was to assess the acute toxicity of B-19704 to juvenile bluegill sunfish, Lepoeis aacrochlrus.
II. IBCnDS
All data and *-he final report vill be naintained either at Baskell Laboratory for Toxicology and Industrial Medicine (Newark, DB) or be archived at the DuPont Records Hanageaent Center (Vilaington, OB)-
III. MATERIALS ARD METHODS
A. Test Substance
The teat substance is described under General Inforaation. 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 deternined.
B. Dilution Vater
Water (Table 1) originated fron the Haskell Laboratory veil.
C. Aniaal Culture
Bluegill sunfish, Lepoais aacrochirus, vere purchased froa either Northeastern Biologists (Rhineback, NT) or Aquatic Research Organises (Haapton, Nfl). Fish vere acdiaated in continuously-flowing, well vater 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 22.69C (definitive study) prior to study start. Pish vere fed newly-hatched, live Artenia sp. (Aquariun Products, Glen Burnie, MD), frozen adult Arteaia sp. (Golden Gate Brand, Kordon Division, Novalek, Inc., Bayvard, CA) and/or Tetraain* flake food (TetraVerke, Geraany) generally twice daily on weekdays and daily on weekends. Fish vere treated vith 2-3 parts per thousand salt (NaCl) on days 0-4 of the 212-day acclimation (rangefinder). Pish vere treated vith three handfuls of salt (NaCl) on day 0, '200 parts per aillion foraalin for one hour on days 1-3 of the 32-day acdiaation (definitive). Sickness, injury and abnoraality vere not present at study start. Aniaals vere identified by labels on aquaria and test vessels.
D. Teat Methods [1]
Seven test concentrations and a control vere used. Noainal test concentrations vere 262, 328, 410. 512, 640, 800 and 1000 ag/L.
Glass aquaria [20 liter; 39.9 (length) x 19.8 (width) x 25.5 ca (height)J
7 Company Sanitized.DoesnotcontainTSCA CBi
containing 10 L of tost solution (approxiaately 13.5-ca liquid depth) yore employed. Positions of test chambers in the water bath used to maintain constant temperature were assigned using random numbers. Ten fish were added to each replicate using randoa nunbers (two replicates per concentration; total 20 fish per concentration). Control fish ranged froa 1.2 to 2.8 ca standard length (naan 2.1 ca) and 0.0451 to 0.524 g vet weight (nean 0.228 g). Control loading was 0.23 g/L at test conclusion. Test solutions ranged froa 21.3 to 22.1*C (wean 21.7C). Pish were not fed approxiaately 24 hours prior to nor during tb'j test. A photoperiod of 16 hours light (312 to 344 Lux) versus 8 hours darkness was eaployed vlth 25 ainutes 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, Yellov Springs Instruments, Yellow Springs, OB), pB (Corning Model 610A, Corning Scientific Instruaents, Medfield, MA) and teaperature (American Society of Testing Materials registered aercury theraoaeter) were aeasured in each replicate before addition of fish at test start, daily, at total fish nortality and/or at test end. Total alkalinity (Bach Cheaical Ccapany, Loveland, CO), EDTA hardness (Bach Cheaical Company, Loveland, CO), and conductivity (YSI Model 33, Yellow Springs Instruaents, Yellov 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 teaperature variations during the 96-hour test.
B. Cheaical Analysis
Chemical analyses of the test substance concentrations in water were not performed because a suitable method was not available.
P. Statistical Methods
The 24-, 48-, 72- and 96-hour LC50s and their 95% fiducial intervals were calculated using probit analysis (2].
IV. RESULTS AMD DISCUSSION
Protocol aaendaents and/or SOP deviations appear in Appendix 1.
A rangefinder was conducted. Noainal test concentrations were 0, 0.1, 1, 10, 100 and 1000 ag/L. Mortality after 96 hours was 0, 0, 0, 0, 0 and 802, respectively.
The definitive study was conducted using noainal B-19704 concentrations of 262, 328, 410, 512, 640, 800 and 1000 ag/L. Ve planned to measure 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.
DuPont BLB 61-94
All chemical and physical paraaeters were within expected ranges (Tables 2-3).
The 96-hour LC50 (Tables 4 and 6) was 634 ng/L with a 9SX fiducial interral of 567 to 725 ng/L based upon noainal concentrations. Behavioral observations are reported in Table 5. Fish eetrics are reported in Table 7.
V. COPCLUSIO*
B-19704 exhibited low toxicity to juvenile bluegill sunfish, Lepoais nacrochirus, in an unaeratedt static test. The 96-hour LC50 was 634 ag/i (95Z fiducial interval of 567 to 725 ng/L) based upon noninal test concentrations.
VI. UTBATOSB CITBD
1. 40 CFR 797.1400. Toxic Substances Control Act. Fish acute toxicity test.
2. Finney, D. J. 1971. Probit Analysis. 3rd Bd., Cambridge Univ. Press. 333 p.
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DuPont BLR 61-94
Static Acuta 96-Hour LC50 of H-19704 To Bluagill Sunfish, Lcpoals aacrochlrus
TABLE 1
Chcaical Characteristics of Haskell Laboratory Veil Vater*
Parameter
Concentration
BOD, ag/L
3
COD, ag/L
<7
DOC, ag/L
0.6
TOC2, ag/L
0.5
Total Kjeldahl N, mg/L <0.2
Aaaonia N, ag/L
<0.1
Turbidity, NTU
<0.5
Phenolics, ag/L
<0.005
Color, apparent Co/Pt3 <5
Solids,
total suspended, mg/L <7
Aluminua, ag/L
<0.2
Antiaony, ag/L
<0.2
Arsenic, ag/L
<0.01
Beryllium, ag/L
<0.01
Boron, ag/L
<0.04
Brralde, ag/L
<0.5
Cadaiua, ag/L
<0.01
Calcium, ag/L
24.5
Chloride, ag/L
6.9
Chroaiua, mg/L
<0.05
Cobalt, ag/L
<0.05
Corper, ag/L
<0.02
Cyanide, ag/L
<0.005
Iron, ag/L
<0.1
Fluoride, ag/L
0.1
Parameter
Concentration
Lead, ag/L
<0.1
Magnesium, mg/L
3.8
MBAS/LAS, mg/L
<0.02
Mercury, ag/L
<0.00020
Nickel, mg/L
<0.05
Nitrite, ag/L
<0.1
Nitrate, ag/L
0.5
Ortho-Phosphate, mg/L <1
Potassium, mg/L
1.8
Selenium, mg/L
<0.005
Silver, mg/L
<0.02
Sodium, mg/L
8.7
Sulfate, mg/L
5.2
Sulfide, mg/L
<0.10
Zinc, mg/L
<0.04
Volatile priority pollutants
Acid extractable priority pollutants Base/neutral priority pollutants Pesticides/PCBs Organophosphate pesticides, ug/L
ND
NDS
ND* ND7
ND*
*Date of saaple collection 12 Nay 1993 unless Indicated otherwise, analyses perforaed at Lancaster Laboratories, Lancaster, Pennsylvania; 2Date of sample collection 23 March 1993 analyses performed at Lancaster Laboratories, Lancaster, Pennsylvania; 3Units based on cobalt/platinum reference; 4None detected; level of detection ranged from < 5 < 100 |ig/L; 5None detected; levels of detection ranged from < 10 to < 25 ug/L; None detected; levels of detection ranged froa < 10 to < 100 yg/L; 7None detected; levels of detection ranged from < 0.01 to 4 ug/L; None detected; levels of detection ranged from < 0.01 to < 0.1 ug/L.
DuPont HUt 61-94
Static, Acute, 96-Hour LC50 of H-19704 To Blueglll Sunfish, Lepoala aacrochlrua
TABLE 2 Vater Chemistry of the- Dilution Vater at Test Start
Total Alkalinity EDTA Hardness (mg/L as CaCOj) (ag/L as CaC03)
79 76
Conductivity (limhos/cm)
165
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mm
il io
Physical and Chemical Characteristics of 3-19704
iTest Solutions
Nominal
Concentration (mg/L)
Dissolved Oxygenb (mg/L)
Control
'02
8.5
8.5
328 8.5
410 8.5
512 8.5
640 8.5
800 8.5
1000
3.5
pH
Control 262 328 410 512 640 800
1000
7.4 7.3 7.4 7.4 7.2
6.9
6.9
7n
8.5 7.8
.5 7.0 8.5 8.0
8.5 7.9
8.5 8.0 8.5 7.9 8.5 8.1 8.5 8.2
7.4 7.4
7.4 7.4
7.4 7.4
7.4 7.5
7.1 7.3
6.9 7.3
6.9 Q
77.3I
Hours
48
72 AB
96 A
7.9 7.5
/.n 7.i>
8.1 7.6
>.9 7.5 8.0 7.7 7.8 7.5 8.1 7.6 8.1 7.7
7.6 7.3
7.B 7.4
7.6 7.3 7.7 7.1
7.7 7.3 7.6 7.0
7.5 6.9 7.6
7.5 7.1
7.5 7.0
7.3 7.0 7.0 7.1
7.2 7.0 7.2 6.9 7.0 6,7 V.O
7.0
7.1
7.1
6.9 7.0 7.0 6.9
6.8
7.4 7.4 7.4 7.4
7.4 7.4 7.4 7.4 7.4 7.3 7.2 7.2
7.3 7.2
7.4 7.3 7.3 7.4
7.4 7.3 7.4 7.3 7.4 7.3 7.2 7.2
7.3 7.3
7.4 7.4 7.3 7.4
7.3 7.3 7.4 7.3 7.4 7.3 7.2 7.2
7.3 7.3
7.3 7.4 7.4 7.3 7.3 7.2 7.2
Company Sanitized.Does notcontainTSCA CBI ''-- I mfiMIMSMliidiMM--1
Static, Acuta, 96-Bour LC50 of H-19704 To Bluogill Sunfish, tappala aacrochirua
DuPont HU 1-94
Static, Acuta, 96-Hour IC50 of H-l>'704 To Bluagill Sunfish, Lapoaia aacrochlrus
TABU 4
Mortality of Bluagill Su fish, ijs nacrochirus.
Exposed to H-19704 |
ijfor 96 Hours
in a Static, naerated
Noainal Concentrations
(Mi/L)
14
Hater Control A* B*
Total
262 A B
Total
328 A B
Total
410 A B
Total
0 0 0
0 10 5
10 10 10
10 20 15
Mortality In Percent Tine (Hours)
48 -- 72
00 00 00 00 10 10 55 10 10 10 10 10 10 10 10 20 20 15 15
96
0 0 0
0 10 5
10 10 10
10 20 15
14
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DuPont BUI Sl-94
Static, Acuta, 96-Hour LC50 of H-19704 To Mungili Sunfish, Lappala aacrochlrua
TABU 4 C0WTIMU1P
Mortality of Bluegill^Sunfish. Legapis aacrochirus. Exposed to H-19704{pBHH9lf3r'^6 Hours in a Statfr, Unaerate<fTest
Mortality in Percent
Noainal
Concentrations (ng/L)
24
512
A* B* Total
30
30 30
640 A B
Total
800 A B
Total
40 40 40
70 60 65
1000 A B
Total
90 90 90
Tioe (Hours) 48 TL
30 30 30 30 30 30
40 40 40 40 40 40
70 70 60 60 65 65
100- 100 90 90 95 95
9g--
30 30 30
40 40 40
70 60 65
100 90 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 HLR 61-94
Static, Acuta, 96-Hour LC50 of B-19704 To Bluegill Sunfisb, Lepoala acroeMm
TABU 5
Behavior of Blueg*ll Sypflah, Lei Is aacrochirus.
Exposed to H-197QA)
tor 96 Hours
in a Stati?' unaeratet fast
Nuaber Shoving Behavior/Total Number 0f Fish Alive
Noainal
Concentrations (ag/L)
24
Tioe (Hours) 48 72
96
Vater Control A B
262 A B
328 A B
410 A B
512 A B
640 A B
800 A B
1000 A B
0/10 0/10
0/10 0/9
0/9 0/9
0/9 0/8
0/7 0/7
0/6 - 0/6
j/3 4*/4
1*/1 1/1
0/10 0/10
0/10 0/9
0/9 0/9
0/9 0/8
0/7 0/7
0/6 0/6
3/3 4/4
__b 1/1
0/10 0/10
0/10 0/9
0/9 0/9
0/9 0/8
0/7 0/7
0/6 0/6
3/3 4/4
__b 1*/1
0/10 0/10
0/10 0/9
0/9 0/9
0/9 0/8
0/7 0/7
0/6 0/6
3/3 4"/4
-- fe
W1
0Lethargic. bAll fiah had previously died.
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DuPont BLR 61-94
Static, Acute, 96-Hour LC50 of H-19704 To Bluagill Sunfiih, Lgponis macrochlrus
H-19704
TABLE 6
Concentrations Causing Mortality to 50Z (LG50) of [ill Sunfish, Lepomls nacrochirus
Time
LC50 (mg/La)
952 Fiducial Limits (mg/L> (mg/L*)
24 Hours 48 Hours 72 Hours 96 Hours
647 634 634 634
575 746 567 725 567 725 567 725
Based upon nominal 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 OA (1-94 Static, Acuto, 96-Hour LGSO of H-19704 To
Blu.gill Sunfloh, Lopoalo nocrochlrua
TABLE 7
Standard Lengths and Vet Weights of Control Fish At Study Bnd
Replicate
Nuaber
A A A A A A
A A A
PA
B B
B B B B
B
B B
B
1 2 3
4 5 7 o
Q
10
1 7 3
A
5
7 8 o
10
Standard Length
(ea)
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
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DuPont BLB 1-94 Sllc,fut* 96-Hour USO of B-19704 To
Blueglll Sunflsh, Lepomla nacrgphtm
PROTOCOL 4MBBBOENIS AND SOP DEVIATIONS PROTOCOL AMENDMENT NUMBER ONE
a. The ratio between test concentrations is 0.8.
nimbi" ' CSiSf" i i 2 f "h0"i'1 b'
10 ,e5t vessels using rsndee
* -
analysis. <TMt*U?Zv .th*
St d' ^ P 18 should be refrigerated prior to Si" of ,he " !*" . e, sill be starved for 24
petfood" 0 " d P*r<iCl,1*'e s" lys" of >h' dilution .ater .ill not be f. Purity and stock checks will not be performed.
S . " d3 i , 2 L 2 S i >
^ 1 " , " .? .ri o.r. " . i , i ; i nf . f^ *
h. Dead fish in the controls do not .ed to be weighed and measured.
i. Test is static (not static reneval).
j. Measure lengths replicates at study
and veights end.
of
all
surviving
fish
in
both
of
the
control
I
19
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DuPont HLR 61-94
Static, Acute, 96-Hour LC50 of H-19704 To Bluegill Sunfish, Lepomis mmcrochirus
APPENDIX 1 (CONTINUED) PROTOCOL AMENDMENTS AND SOP DEVIATIONS CONTINUBD
PROTOCOL AMENDMENT NUMBER TWO
Ve planned to measure H-19704 concentrations in each test solution. Samples for analysis were taken and the analyses were attempted. The analyses failed and preliminary research suggested that it was rot cost effective to develop a new method. The samples were therefore discarded and nominal concentrations reported. Since the purpose of this study was tc provide acute mortality data for designing a bioconcentration test, this procedure was considered acceptable.
In 1994 Haskell Laboratory adopted a new system 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 na of the sponsoring business is DuPont Elastomers (the previous
name was Polymers 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.
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