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Du Pent BLR 395-91
Study Title
OPi STATIC. A^EJ9^H^1R LC50
TO FATHEAD HlHNOVS (Piaephales pgjftfelae)
D^ta Reguireaent
United States EnvironBental Protection Agency Environmental Effects Testing Guidelines Subdivision EG-3
; Author
Kevj|nN. Baer, PhD.
i
Study Completed On
September 4, 1991
i
Performing Laboratory
Haskell
I. E.
du Font de Honours and Company
Laboratoryjr for i'TTooi xicology and Industrial Elktoonn RRooaidd,, Post Office Box 50
Newacrk, Delaware 19714
Medicine
Laboratory Project ID Haskell Laboratory Report No. 395-91
Page 1 of 14
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^J'lPont HUl 395-91
QUALITY ASSURANCDEOCUHENTATIOM
STUDY;
96-HOUR LC50 OF FATHEAD MINNOVS
promelas)
AUDITS;
Items Audited
In-life Observations
Protocol, Records and Final Report
Audit Dates |5/15/91
i
J7/22,23/91
DATE FINDINGS REPORTED TO w|HAGEMEANMDT STUDY DIRECTOR; 7/23/91
Reported by:
Joseph C.
Senior Auditor
Quality Assurance
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'Date
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Good Laboratory Practice jSti^aBSl^'., ..:.;.':, --n/..'.
2
. . . . . . . . . Quality Assurance DoeuB-intattm
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3
..,...,. .......... Table of Contents ......,.......... 4
I. SuDoary
5
............................ II. General Information
6
...!.........,.......... III. Purpose
7
......... I.................... IV. Records
7
............................. V. Materials and Methods
8
A. Test Material
8
...................... B. Dilution Vater
8
........................ C. Animal Culture
8
....................... D. Test Methods
8
....................... E. ........................ F.
CShteamtiisctailcAal Mnethaodls ysis......................
9 9
..................... I. Results and Discussion
9
..................... VII. Conclusion
9
.
........................... VIII. Literature Cited
10
........................ IX. Tables
11
1. Chemical Characteristics of Haskell Laboratory Veil Water . . 11
............................. .... 2. Water Chemistry of the Dilution Water atJTest Starts.
12
3. Physical and Chemical Characteristics ofglHmf^Test
Solutions
.1
13
4. Observed Mortalit^^^Bathead Minnows (Pinephales promelas)
...... ................... Exposed toilHB--Rlfor 96 Hours in a Static,
Aerated Test ....' ................... 14
. co^nys.^-.^'"1"'"*^*'181
I,
not cuiteiy toxic t< j^lBteK
proaelas) in a'^S-hour, aerate-d".";
646, 1660, 1800, 3000, and 5000 concentrations vas greater than
itatl< tBic. I The 96-h
g/1., the h
'If^ . <>','
Du Font BLR 395-91
Binnovs (P^ephlcB
test concdntratloncit vexe
> based on nominal
concentration tested.
<"* y ^^??J V. TtoyJB^leF Technician
^^^^ ^
R. Scott --ria'-n
Tp"hnician !
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Robert V. Peterson Technician i
Study
Director!
Wallace E. Eessicks
| Technician ' y
^^^^^ Ke^inH. Ba|er, Ph.D.
Research To^icologist Environmental Sciences
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^ THIS REPORT IS APPROVED FOR ISSUE.
ji^ ^.y ^L
Kevin N. :-'a|er^ Ph.D. /study Din*a'tor
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Study Schedules
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Study IiiUat*lft I 8^e?lBeiHi|L.^!il Experimental TS " Study e@w|i|;<8tiny
Notebooks!
t 5/14/91
tei 5/15/91 ion Date: 5/19/91 s 9/4/91
Distributions
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'Company
V. HATBRIALS AHD HBIBOD5
Du Pent HLR 395-91
A. Teat Material
Source, eoaposition, and purity of th t-eKt
General Inforoation. The appropriate quantity dispensed into 4-L of dilution water (see r
concentration. Stability and solubility e
not determined.
I
B. Dilution Vater
|
described under
|vas measured and
each test in dilution water vere
Vater (Table 1) originated froa the Haskell Laboratory well.
C. Animal Culture
|
Fathead minnows (Pimephales broaelas) were reared in continuously flowing veil vater at Haskell Laboratory^Adult fish resided in 38 L aquaria at approximately 258C and were fed frozen brine shrimp (Artemia sp., Golden Gate Brand, Korden Division, Novaiek, jinc., Hayward, CA) generally twice daily, once on Saturday, and not on Sundays. | Eggs were deposited on polyvinyl chloride tiles coated with sand. Tiles vijth eggs were aerated and incubated at approximately 25C in a water bach. Larvae were reared in 22 L aquaria at approximately 22C and were fed newly hatched Artenia generally twice daily on weekdays and daily on weekends. jJuveniles were held in a 22 L aquaria at approxiaately 22 C and were fed newly hatched Arteaja and/or frozen brine shrimp generally twice daily on weekdays and once on Saturday and Sundays. No treatment of the fish for-diseases was required during the holding period. Sickness, injury, and abnormality were not observed. Animals were identified by labels on aquaria and test vesjsels.
D. Test Methods [1]
Five test concentrations and i water control were used. Nominal test concentrations were 648, 1080, 1800, 3000, and 5000 mg/L. The fathead minnows ranged from 1.5 to 2.4 cm in staniard length (mean 1.9 cm) and 0.029 to 0.16 g in weight (mean 0.092 g). The fathead minnows were 85-86 days old at test start.
Glass aquaria (5-L) containing 4 L of test solution (approximately 12-cm liquid depth) were employed. TWO! replicates of each test concentration with ten fish per relicate were used. Fish were added to the test concentrations using random numbers and were not| fed for 48 hours prior to the test nor during the test* Mean loading for controls at test conclusion was 0.23 g/L. Test
solutions were held between 20.7 tmd 21.6BC (mean 21.2C) and were unaerated, until at approximately 72 hours wnen low dissolved oxygen levels necessitated the use of gentle aeration (approximately 120 bubbles/minute). A photoperiod
of 16 hours light (215 Lux) versu^ 8 hours darkness was employed.
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Du Font BLR 395-91
Dissolved oxygen (TSI Model
Springs, OH), pB (Corning N^H.
;nstrumnfi Yellw fie instruRents, Medfield,
MA), and temperature (Aiieri^ii^'i
:erial8 registered WBrcury
stea therBoaeter) were lieasu'lNI control at the beeleming of the.
itions and the water
i of fish) and daily
thereafter* Total alfciallnity
il Company, Lovelcnd, CO),
EDTA hardness (Hach kit, Bach
and, CO), and conductivity
(YSI Model 33, Yellow Springs I:nstnwent, Yellow Sprlnga, 08) of the dilution
water vere measured at the beg: ng of the tesit. Total organic carbon (Model
700 01 TOC analyzer) was iieasuc^d in the dilution water at 48 hours.
B. Cheaical Analysis
Chemical analysis of the te^t material was not conducted.
P. Statistical Methods
!
.
The 96-hour LC50 p--i its associated confidence interval could not be
calculated because mortality
not observed at any test concentration.
VI. JffiSULTSAND DISCUSSION
i
A 96-hour rangeflnding stud;r was conducted using the following
concentrations; water control, : .0, 10, 100, and 1000 ng/L. observed at any test concentrat: on.
Mortality was not
Nominal^HPHH^lconcenttationswere 649, 1080, 1800, 3000, end 5000
ng/L. All test concentrations (excluding control) were cloudy throughout the
testing period.
All chemical and physical parameters (Tables 2 and 3) were within expected ranges, except dissolved oxygen;in replicate A of the 5000 mg/L test concentration at 72 hours. The(dissolved oxygen value (5.3 mg/L) dropped slightly below 60X of the theoretical saturation value of 9.0 mg/L at 21C. As
a result, all test concentrations were gently aerated beginning at 72 hours
(approximately 120 bubbles/minute). This apparent decrease in dissolved oxygen
was not considered biologically'significant based on the lack of observed mortality. No mortality or sublethal effects were seen in any test concentration (Table 4). The 9^-hour LC50 based on nominal concentrations was greater than 5000 mg/L, the highest concentration tested.
^1.1. CONCLUSION
Q^BIHHQwas not acutely toxic to juvenile, fathead minnows (Pimephales
promelas) in a 96-hour aerated, static, test. The 96-hour LC50 based on nominal concentrations was greater than 5000 mg/L, the highest concentration tested.
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Chemical Characterint
Du Pent BLR 395-91
l-H
^
^osiW Veil Water*
Parameter
Concentration
Feraceter
Concentration
BOD2, ag/L
COD, mg/L
DOC3, ig/L
TOC, Cg/L
Kjeldahl N, og/L
Aminonia N, g/L Turbidity, HTU
Phenolics, mg/L Color, Co/Pt Solids
total suspended,
Aluminum, nsg/L
Antimony, mg/L
Arsenic, mg/L Beryllium, mg/L Boron5, mg/L
Cadmium, mg/L
Calcium, mg/L Chloride6, mg/L
Chromium, mg/L
Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron6, mg/L Fluorlde6, ag/L
<2 13
1.5 <0.5
<00..02|,5
<1.0. <0.050
<5 !
mg/L
<2
<0.130 <0.050 <0.0l0 <0.0)1 <o.ozo <0.002 25.0!
6.3!
<o.qio <o.ozo <0.010
<O.OZ5
0.053 <0.1
Leads. ift/L
Magnesium, mg/L
MBAS/LAS Bg/L
Mercury, ag/L
m^wii ilg/L
ttrtt<, Bg/L Nitrate6, mg/L
Phosphate6, mg/L
PotassiuD, mg/L Seieniua, og/L
Silver, ag/L
Sodiin, mg/L
Sulfate6, rog/L Sulfide, mg/L Thallium, mg/L Zinc8, mg/L Volatile priority pollutants Acid extractable priority pollutants Base/neutral
priority pollutants
Pesticides/PCBs
Organophosphate
pesticides7, ug/L
<0.005
3.'i <0.05
<0.00020 <Q^2Q <0.1
1.6 <0.1
1.8
<0.005 <0.310
8.1 4.8
<0.05 <0.01 <0.020
ND8
ND8
ND'
MD
<0.50
^Date of analyses 17 October 1990 unless indicated otherwise, analyses
performed at Environmental Testing and Certification Corporation, Edison, New Jersey; ^Date of analyses 13 December 1990, analyses analysis performed at Environmental Testing and Certilfication Corporation, Edison, New Jersey; 'Mean value; Date of analyses 17 Hay 1991, performed at Du Font Engineering; Test Center, Newark, Delaware; ^eldw the Published Method Detection Limit (BHDL);
^ate of analyses 9 November 1990, analyses performed at Du Font Chemicals,
Jackson Laboratory, Deepwater, iHew Jersey; ''Date of analyses 23 October 1990, analyses performed at Hazleton^Laboratories, Inc., Madison, wi; SNone detected.
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Physical and
Du Pent BLR 395-91 of
Tiae (Hours)
Water Control
A" &
648 A" 5"
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^M Noninal Gonc^ntrations
1080 A" B-
1800 A* *
3000 A" B"
5bu0
A B
Dissolved oxygen (ng/L)
Start
24 48 72 96
8.8 8.8 7.9 7.7 7.7 7.7 7.6 7.5 8.2 8.0
8.8 8.81 7.7 7.4! 7.2 7.3; 7.2 7.3| 7.7 7.6;
8.8 8.8 7.4 7.6 7.1 6.9 6.9 6.8 7.3 7.2
8.8 8.8 7.2 7.7 6.4 7.1 6.7 6.8 7.6 6.9
8.7 8.7 7.6 7.5 6.6 6.4 5.9 6.2 6.3 6.7
8.5 8.6 7.8 7.7 6.7 6.0 5.3 5.5 6.1 6.7
pH
Start
24 48 72 96
7.3 7.3 7.1 7.2 7.2 7.4 8.1 8.1 8.3 8.3
7.3 7.3| 7.3 7.4 7.2 7.3 7.3 7.3 7.4 7.4 7.4 7.4 8.2 8.1 8.1 8.1 8.2 8.2 8.1 8.1
7.4 7.4 7.3 7.3 7.4 7.5 8.0 8.1 8.1 8.0
7.4 7.4 7.3 7.3 7.5 7.5 8.1 8.0 8.0 8.0
7.3 7.4 7.3 7.3 7.5 7.5 7.9 7.9 8.0 7.9
A and B refer to replicates ^ith ten fish each.
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D<* Pottt aLR 39S-91
Observed
m w ^ i . . %:^ 1 Mortality of Pa vWi^^^^f '^rWv^W^f^ 'w^ffEpy^wSB ^^^^^^PBBHIHiB^B
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for 96 Romp n ^..Sttl^L'^alfeN^esf
Concentrations (Bg/L)
Percent Cirulative Observed Mortality"
24 Hours
: 48 Hours
*
72 Hjpurs
96 Hours
Ab
B"
Ab
Bb
A*
Bt
Ab
Bb
Water Control
00 00 00 648
00 00 00 1080
00 00 00 lib
1800
IBly
00^00 00 3000
0000 00 ,00 0000 5000
All fish appeared normal.
<>A and B refer to replicates! vith ten fish each.
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