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Du Font HLR 592-92
TRADE SECRET
StutfyTite STATIC, ACUTE, 96-HOUR LC50 OF H-19.517
TO FATHEAD MINNOWS. Pimephales pmmelas
Data Requirement Organisation for Economic Co-Operation and Development (OECD)
Guideline for Testing Chemicals: 203
Author Keith B. Pieison, Ph.D.
StjdyCompleted On Novembers, 1992
Performing Laboratory E. I, du Pont de Nemours and Company (-tasked Laboratory for Toxicology and industriai Medicine
Efkton Road, Post Office Box 50 Newark, Delaware 19714
Lstrorstonf ProjecLID Haskell Laboratory Report No. 592-92
Page 1 of 19
^^s A O09
QtiSfff9 ,s^
Du Pont HLR 592-92
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to EPA FIFRA and TSCA Good Laboratory Practice Standards (40 CFR 160 and 40 CFR 792). OECD Principles of Good Laboratory Practice (C(81)30(Final), Annex 2), and MAFF Japan Good Laboratory Practice Standards (59 NohSan No. 3850) with the two exceptions noted below. Solubility and stability of the test substance in dilution water and under storage conditions were not determined. Concentrations of the test substance in dilution water were measured, but these measurements were not made in accordance with Good Laboratory Practice Standards. Protocol amendments and/or SOP deviations appear in Appendix I. Areas of noncompliance (if any) are also documented in the study records. No deviations existed that affected the scientific validity of the study.
Subr.-'itter:
E. I. du Pont de Nemours and Company
Sponsor:
Du Pont Chemicals E. I. du Pont de Nemours and Company
STUDY DIRECTOR:
rW^ 4 -A^s-^
Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences
^ A/fff/ WA. Mo/Day/Year
otco^n-^081
co^s^0065"010
2
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Du Font HLR 592-92
TABLE OF CONTENTS
Page
Good Laboratory Practice Statement...................................................^
Table of Contents........................................................................3
General Information......................................................................4
Summary.................................................................................6 Quality Assurance Documentation .................................>....................7
I. Purpose.........................................................................8
.I. Records.... ....................................................................8
III. Materials and Methods ..........................................................8
A. Test Substance..............................................................8
B. Dilution Water...............................................................8
C. Animal Culture ..............................................................8 D. Test MicealthAnoadlyss.is...............................................................9
E. Chem Statistical
.............. ..........................................1.90
F.
Analysis........................................................
IV. Results and Discussion....................................................... 10
V. Conclusion.................................................................... 10
VI. Literature Cited ............................................................... 10
VII. TaCbhleemsi.c.a.l..C..h.a.r.a.c..te..r.is..ti.c.s..o..f.H..a.s..k.e.l.l..L.a.b..o.r.a.t.o..r.y..W..e.l.l.W...a.t.e.r..............11
1. 2.
Measured Concentrations of H-19,517 in a
96-Hour, Static, .............11
WUnaateerraCtehoemTiesstrty .o.f..th..e..D.i.lu..ti.o.n..W...a.t.e.r..a.t.T..e.s.t..S.t.a.r.t.......................12
3. 4.
Physical and Chemical Characteristics of H-19,517.T..e.s.t..S..o.l.u..t.io..n.s..........1134
5. Observed Mortality of Fathead Minnows. Pimephales promelas,
Exposed to H-19,517 for 96 Hours in a Static, Unaerated Test.......... 16
6. Observed Effects of H-19,517 on Fathead Minnows, Pimephales
7. Hpr-o1mS.e5la1s7. Cfoornc9e6nHtroautirosnisn Ca aSutsaitnicg, MUnoartearlaittyedtoT5e0%st.(L..C..5.0.)..o..f ........... 17
Fathead Minnows, PimephaSes promelas, at Specific Time Intervals.... 18
VIII.
A pPproetnocdoil xA.m. .e. .n.d. .m. .e.n.t.s. .a. .n.d. .S. .O. .P. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .
19 19
1.
Deviations..............................
.....TSCACBI
o^s^.o--------
3
Du Pone
GENERAL INFORMATION
HastetLMimbsc
EUjntMI Composition:
H-19.517
Contaminants: CAS_BegIstiyJJumber:
Not supplied by sponsor None avaifabie for this mixture
Seonsfijr;
Du Font ChemteaSs E. 1. du Pont d Nemours and Company Wilmington. Delaware 19898
Srf^."-""-"'-"TM"08'
4
Company Saffilized. &sss r<os Gonsas" TSCA CB4
Du Pom HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS. Pimephales pmnBlas
SUMMARY
H-19,517 exhibited moderate toxidty in a 96-hour, unaerated, static, acute test using fathead minnows, Pimephales promelas. Nominal H-19,517 concentrations were 0.0.409,1.02,2.59.6.41.16.0,40.0. and 100 mg/L H-19,517 measured concentrations at 96 hours were 0.00,0.16,0.55,1.88,4.03, 7.46,15.78, and 48.11 mg/L. The 96-hour LC50 based on measured concentrations at 96 hours
was 4.72 mg/L with a 95% confidence interval of 3.55 to 6.02 mg/L.
WorkBy:
^ ^ 2 ^ ^ Robert W. Peterson, B. A.
Toxicology Associate
/^//,/? ^
Mo/Day/Year
Work By:
_Je^ffrbeyj_T.LTurn-e^r,--Sr.
Toxicology Associate
fo/j'/y^
Mo/Day/Year
THIS REPORT IS APPROVED FOR ISSUE.
AwfL ^ ftc^'^
Keith B. Pierson, Ph.D. Research Toxicologist and Study Director Environmental Sciences
^ fifoV ^2.
Mo/Day/Year
Company Sanitized. Does not eanSain TSCA CB1 6
Du Pom HLR 592-92 QUALITY ASSURANCE DOCUMENTATION
:ates of Inspection:
Conduct - 7/23/92; 8/5/92
Records, Report(s) -10/27 - 30/92
ridings reported to:
Study Director - 10/30/92 Management- 11/6/92
Reported by:
i-.^. u -A-1
Deborah A. Vick Quality Assurance Auditor
i'1^1^
Date
Company Sani88zec8. Does not contaid TSCA CBI
Du Pom HLR 592-92
I. PURPOSE The goal of this study was to assess the acute toxicity of H-19,517 to fathead minnows, Pimephafes promelas.
II. RECORDS
All data and Toxicology
the
final
report
will
be
maintained
eithei
at
Haskeli
Laboratory
for
Du
Font
and Industrial Medicine (Newark, Delaware) Records Management Center (Wilmington,
or
be
archived
at
the
Delaware).
Hi. MATERIALS AND METHODS
A. Test Substance
The test substance is descnoed under General Information. was added to dilution
The 13st substance
stock solution.
The
water .see below) and stirred for 30 stock solution was added to dilution
minutes
to
create
a
volumes to make individual test solutions.
water in appropriate Stability and solubility of'H-19,517
dilution water and under sto.-age conditions were not determined.
in
B. Dilution Water
Water (Table 1) originated f'om the Haskeli Laboratory well. C. Animal Culture
Fathead Minnows, PimephLies promelas, were reared in continuously flowing
well water at Haskeli Laboratory. Adult fish resided in 39 L aquaria at
approximately 25C and Gate Brand, Kordon
were
fed
frozen
brine
shrimp
(Artemia
sp..
Golden
once
on
Saturday
Division. Novaiek, Inc., Hayward, CA) generally and once on Sunday. Eggs were deposited
twice
daily,
chloride tiles coated with sand.
on polyvinyl Tiles with eggs were aerated and incubated at
approximately approximately
25C 21 C
in a
and
water bath. Larvae were were fed newly hatched
reared
in
39
L
aquaria
at
Products,
Glen
Burnie,
MD)
generally
twice
daily
Ailemiasp. (Aquarium on weekdays and daily
weekends. 39 L
Juveniles
were
held
either
in
272-L
circular,
fiberglass
tanks
on or in
21C galnasdswaeqrueafreiad (n2e3w-clymhdaetpchthe;d5A0rt(eLm) xia2s5p(.Wan) dx /o31r fcromze(Hn )b) raint eapsphrroimxipmately
generally twice daily on weekdays, once on Saturday and once on Sunday.
bSyiclkanbeeslss, oinnjuarqyu, aarniad aanbdnotersmtavlietysswelesr.e not observed. Animals were identified
SasiteeA Ows ml -wntsm TSCA CBJ
Du Font HLR 592-92
D. Test Methods [1]
Seven test concentrations and a water control were used. Nominal concentrations were 0, 0.409. 1-02, 2.59, 6.41,16.3, 40.0, and 100 mg/L. Control fish ranged from 1.7 to 2.4 cm standard length (mean 1.9 cm) and 0.11 to 0.25 g wet weight (maan
0.18g).
Erienmeyer glass flasks [ 4.4 liter; 21 (base) x 35 (height) cm] containing 4.4 L of test solution (35-cm depth) were employed as test chambers. Each flask was
totally filled to minimize head space and was covered with Saran Wrap to
prevent loss of potentially volatile test substance components. The position of
test chambers in the water bath was determined by use of random nu
'
?rs.
Ten fisft were added to each replicate using random numbers (one replicate per concentration, total 10 fish per concentration). Control loading was 0.40 g/L at
test conclusion. Test solutions ranged from 20.7 to 21.5C (mean 21.0C). Fish were not fed 27 hours prior to nor during the test. A photoperiod of 16 hours light
(172 to 215 Lux) versus 8 hours darkness was employed with 25 minutes of transitional light (2 to 3 Lux) preceding and following the 16 hour tight interval.
Mortality and behavioral effects were noted da:.y.
Dissolved oxygen (YSI Model 54A, Yellow Springs Instruments, Yellow
Springs, OH) and pH (Corning Model 61 OA, Coming Scientific Instruments,
Medfield, MA^ were measured in the water control and all test concentrations before fish were added at the beginning of the test and daily thereafter. Temperature was measured (American Society of Testing Materials registered mercury thermometer) in the water control and all test concentrations before fish
were added at the beginning of the test, daily thereafter, and at the end of the test. Total alkalinity (Hach kit. Hach Chemical Company. Loveland, CO), EDTA hardness (Hach kit, Hach Chemical Company, Loveland, CO), and conductivity
(YSI Model 33, Yellow Springs Instruments, Yellow Springs. OH) of the dilution water were measured at the beginning of the test. A continuously-recording
thermometer in a control vessel was used to check for temperature variation
during the 96-hour test.
,
E. Chemical Analysis
Analyses were conducted after the test had ended at the Du Font Jackson Laboratory at Chambers Works, Deepwater, NJ. Samples and standards were completely combusted in a hydrogen/oxygen Wickbold Torch. The resulting water was then analyzed for total fluoride concentration using an Orion fluoride ion selective electrode. Concentrations were then calculated from the standards using either a three or four point calibration curve.
Company Sanitized. Does not contain TSCA CBi 9
Du Pont HLR 592-92
F. Statistical Analysis The 24-. 48-, 72-, and 96-hour LC50s we'e calculated by using the moving average angle technique [2].
IV. RESULTS AND DISCUSSION A rangefinding test with five fish per concentration (4.4 L Erienmeyer flask filled to
the top) was conducted with a water con?-3l and test concentrations of 0.1, 1, 10, 100, and 1000 mg/L. Mortalities were 0. 3. 0, 60,10U, and 100%, respectively. Two definitive studies were conducted. Cata from the first were discarded because of tow dissolved oxygen concer--ations, and data from the second are reported herein.
Nominal concentrations in the definitive test were 0, 0.409, 1.02, 2.59, 6.41, 16.0, 40.0, and 100 mg/L Measured concentra:ions at 96 hours were 0.00, 0.16, 0.55, 1.88, 4.03, 7.46, 15.78 and 48.11 mg.L ^he discrepancy between the measured and nominal concentrations (Table 2) ma< stem from the use of a stock solution in which the test substance was not totally c.ssolved. Al! chemical and physical parameters in the final definitive test (Ta^ es 3 and 4) were within expected ranges with one exception. The dissolves oxygen concentration in the control and 4-03 mg/L test solutions on day 4 d'coped to 58 and 56% of saturation, respectively. This observation was not c' biological significance since fathead minnows can survive at much lower diss^ved oxygen concentrations for extended periods. Behavioral obsen/a1ic"s appear in Table 6. The 96-hour LC50 based on measured concentrations at 96 hours was 4.72 mg/L with a 95% confidence interval of 3.55-6.02 mg.'L (TDICS 5 and 7).
V. CONCLUSION
H-19,517 was moderately toxic to fathees minnows, Pimephales promelas. in a 96-hour, static, unaerated test.
VI. LITERATURE CiTED 1. Organisation for Economic Co-Opera: on and Development (OECD).
Guideline for Testing Chemicals: 203 4 April 1984.
2. Pettier, W. H. and C. I. Weber, Eos. 1985. Method3 for measuring the acute toxicity of effluents to freshwater and marine organisms. EPA/600/4-85/013. United States Environmental Protect 3n Agency, Environmental Monitoring and Support L aboratory, Cincinnati, Cnio. Appendix E.
Company Sanitized. Does not contain TSCA CBI
10
Du Font HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS. Pimephales promelas
TABLE 1
Chemical Characteristics of Haskell Laboratory Well Water1
Parameter
Concentration
BOD, mg/L COD, mg/L DOC. mg/L
TOC2, mg/L Total Kjeldahl N, mg/L Ammonia N, mg/L Turbidity, NTU
Phenolics, mg/L Color, apparent Co/Pt7 Solids
Total suspended, mg/L Aluminum, mg/L Antimony, mg/L Arsenic, mg/L Beryllium, mg/L Boron4, mg/L
Cadmium4, mg/L Calcium, mg/L
Chloride3- mg/L Chromium, mg/L Cobalt, mg/L
Copper, mg/L Cyanide, mg/L Iron3, mg/L
Fiuoride3, mg/L
<2 <10
1.4 2.9 0.8 <0.2 <1.0 <O.G50 <5
<4 <0.100 <0.060 <0.010 <0.001
0.020 <0.002 24.3 6.2 <0.010 <0.020 <0.010 <0.025 0.001
<0.1
Parameter
Lead. mg/L Magnesium, mg/L MBAS/LAS, mg/L Mercury, mg/L Nickel, mg/L Nitrite3, mg/L
Nitrate3, mg/L Phosphate3, mg/L Potassium, mg/L Selenium, mg/L Silver, mg/L Sodium, mg/L Sulfate3, mg/L Sulfide, mg/L Zinc, mg/L Volatile priority pollutants Acid extractable priority pollutants
Base/neutral priority pollutants Pesticides/PCBs Organopnospnate pesticides5, ng/L
Concentration
<0.075 3.7
<0.05 <0.00020 <0.020
<0.1 1.9
<0.1 1.7
<0.005 <0.010
8.2 5.0 <0.10 <0.058
ND6
ND6
ND6 NO6
<0.50
^Date of sample collection 26 May i992 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersay; 2Date of sample collection 1 Juiy 1992, performed at Du Pont Engineering Test Center, Newark, Delaware;
^ate of collection 10 June 1992, analyses performed at Du Pont Chemicals, Jackson
Laboratory, Deepwater, New Jersey; ^Below the Published Method Detection Limit (BMDL); SAnalyses performed at Hazleton Laboratories. Inc.. Madison, WI; SNone detected; ^Units based on cobalt/platinum reference.
Company Sanitized. Does not contain TSCA CBI
Du Font HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19.517 TO FATHEAD MINNOWS, Pimephales promelas
TABLE 2
Measured Concentrations of H-19,517 in a 96-Hour, Static, Unaerated Test
Nominal Concentration (mg/L)
Water Control 0.409 1.02 2.59 6.41
16.0 40.0 100
Measured Concentration (mg/L)
StartEnd
0.00 0.40 0.76 2.38 3.87 9.86 18.85 53.76
0.00 0.16 0.55 1.88 4.03 7.463 15.783 48.113
''Samples taken at time of total mortality.
CompanySanjfeed.^DW^c_wsarnTSCACW
12
Du Pont HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19.517 TO FATHEAD MSNNOWS, Pimephales promelas
TABLE 3 Water Chemistry of the Dilution Water at Test Start
Total Alkalinity (mg/L as CaCOa)
79
EDTA Hardness (mg/L as CaCOa)
77
Conductivity (umhos/cm)
170
Company SanitBed.ooes oes nnoottc.ont, ain TSCACBi
13
Du Font HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas
TABLE 4
Physical and Chemical Characteristics of H-19,517 Test Solutions
Measured Concentration
(mg/L)
^
D-^olved Oxvoen. rrig-"/L
HgO Control
0.16 0.55 1.88 4.03
746
15.78 45.11
DH
HaO Control
0.16 0.55 1.88 4.03 7.46 15.76 46 1 1
Time OHr 24 Hr 48 Hr 72 Hr 96 Hr
8.4
7.0
6.4
6.0
5.2
8.4
7.0
6.4
6.1
6.0
8.4
7.2
6.7
6.3
5.7
8.4
7.2
6.6
6.3
5.4
8.5
7.4
6.9
6.4
5.0
8.5
7.7
a
a
a
8.4
8.0
a
a
a
8.4
8.0
a
a
a
7.6
7.4
7.4
7.4
7.0
7.6
7.4
7.4
7.4
7.0
7.6
7.4
7A
7.4
7.1
7.6
7.4
7.4
7.4
7.1
7.7
7.4
7.4
7.4
7.1
7.6
7.4
a
a
a
7.6
7.5
a
a
a
7.6
7.5
a
a
a
leading was not made due to total mortality.
^^^Tsc^e,
14
Du Poni HLR 592-92
STATIC, ACUTE 56-HOUR LC50 OF H-19,517 TO FATHEAD MiNNOWS, Pimephales promelas
TABLE 4, Continued
Physical and Che~ :ai Characteristics of H-19,517 "ssi Solutions
Measured Concentration
(mg/L)
Time 0 f-- 24 Mr 48 Hr 72 Hr 96 Hr
Temperature. C
HgO Control 0.16 0.55 1.88 4.03 7.46
15.73 48.11
20" 20-
20.5
209 20" 20= 20=
20 :
20.9 20.9 20.9 20.9 20.9 20.9 20.9 20.9
20.8 20.8 20.8 20.8 20.8
a
a
a
21.5 21.5 21.5 21.5 21.5
a
a
a
21.4 21.4 21.4 21.4 21.4
a
a
a
'Reading was not made due ';
mortality.
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I 5
Du Pont HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS, Pimephales promelas
TABLE 5
Observed Exposed
Mortality of
Fathead
Minnows,
Pimephales promelas,
to H-19,517 for 96 Hours in a Static, Unaerated Teest
Measured
Concentration (mg/L)
24 Hours
Cumulative Mo rtality in Percerit'1
48 Hours
72 Hours
96 Hours
Water Control
O.lf
0.55 1.88 4.03 7.46 15.78 48.11
0 0 0 0 0
100
.
100 100
0 0 0 0
20 100 100 100
0 0 0 0
20 100 100 100
0 0 0 0
20 100 100 100
s'mere were ten fish per test concentration at test start.
16
Du Font HLR 592-92
STATIC, ACUTE, 96-HOUR LC50 OF H-19,517 TO FATHEAD MINNOWS. Pimephales promelas
TABLE 6 Observed Effects foofr H96-1H9.o5u1r7s oinn aFaStthateiac,d UMnianenroawtesd, PTeimstephales p'omelas,
Measured Concentrations
(mg/L)
Water Control 0.16 0.55 1.88 4.03
7.46 15.78 48.11
Number of Fish With Effect
24 Hours
48 Hours
72 Hours
0/10 0/10 0/10 0/10 13/10
1b/10
C
C
C
0/10 0/10 0/10 0/10
Z^'Q
C C C
0/10 0/10 0/10 0/10 3^8
C C C
96 Hours
0/10 0/10 0/10 0/10 3^8
C C C
''Lying on bottom of the test vessels. "Partial loss of equilibrium.
Total mortality.
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17
Du Font HLR 592-92
TSOTAFTAITCH, EAACDUTMEI,N9N6O-HWOSU. RP'LmCe5p0haOleFsHp-r1o9m,5e1la7s
TABLE 7
H-19,517 Concentrations Causing Fathead Minnows, Pimephafes
Mortality
to
50%
(LC50)
of
promelas, at Specific Time Intervals
Time
LC50
(rng/L3)
95% Confidence Limits (mg/L3)
Slope
Y-intercept
24 Hours
5.48
4,24
7.09
N/Ab
N/A
48 Hours 4.72 3.55 6.02 N/A N/A
72 Hours 4.72 3.55 6.02 N/A N/A
96 Hours
4.72
3.55
6.02
N/A
N/A
-'Based upc" measured concentrations at 96 hours.
^ot applicable.
angle technique
Slope and Y-intercepr not calculated by the mo. ng average
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18
Du Pom HLR 592-92
STATIC, ACUTE. 96-HOUR LC50 OF H-19,51" TO FATHEAD MINNOWS. Pimephales promel&s
APPENDIX 1 PROTOCOL AMENDMENTS and SOP DEVIATIONS PROTOCOL AMENDMENT NUMBER ONE
a. It is not necessary to weijh fish and determine test chamber lose ng prior to
test start.
b. Stability and solubility of the test substance in dilution water anc jnder storage conditions will not be determined.
"' c. Samples will be taken from each replicate at test start and end
analysis of the test substance concentration.
chemical
d. At test end surviving fish will be sacrificed according to aquatic 5 DP AQ029P-002.
e. Fish will be starved for 24 hours prior to test start. PROTOCOL AMENDMENT NUMBER TWO
a. The test substance concentrations were changed to facilitate the >veighing
process and will be: 0.409, 1.02, 2.59, 6.41, 16.0, 40.0, and 10C ig/L
SOP DEVIATIONS
a. Temperature of water bath was not recorded. b. Temperatures and times to equilibria at individual test containers .vere not
recorded.
c Maintenance was not performed on balance LI 642 when calibre: an was out of Standard Operating Procedure range.
d. Which pH calibration buffers were used was not recorded durinc :H meter calibration on 7-25-92, 7-26-92, 7-27-92, and 8-5-92 through 8^-92.
e. iSntusdoymeDitreescttosrolnuotitonno. tified on 8-9-92 of low dissolved oxygen cc";entration
CompanySanitized.
Does not contain TSCA CBS
19