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Du Font HLR 457-91
Study Title
STATIC, ACUTE, 48- AND 64-HOUR EC50 O^II^JUr10 Paphnia aagna
Data Requirement Organisation tor Economic Co-Operation and DevelopfBent (OECD)
GuiHeifne for' festTng ChemicaTsY 2'02'
Author Kevin N. Baer, Ph.D.
Study Completed On June 26, 1991
Performing Laboratory
Haskell
E. I. dii Font de Nemours and Company
Laboratory for Tovicolc-gy and Industrial Elk ton Road, Post Office Box 50 Newark, Delaware 197U
"sdlcins
Laboratory Project ID Haskell Laboratory Report No. 457-91
Page 1 of 15
Company SanU-aed.1^
-.tgTSCACa
mS PACT USBVID FOR SHCIFIC COURT
Du Font BLR 457-91
1
Du Font BLR 457-91
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to United Stales Environmental Protection Agency Good Laboratory Practice Regulations for FIFRA (40 CFR 160). Areas of
non-compliance are documented in the study records. No deviations existed that
significantly affected the validity of the study.
Submitter: Sponsor:
E. I. du Pont de Nemours and Company
Du Pont Chemicals
Study Director:
//i'^
Kevin N.
B/a/e.r,';^P^h.-D.,
Research Toxicologist
Environmental Sciences
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Company Sanitized. Does ncBc ariaisTSCAC
Du Font HLR 457-91
QUALITY ASSURANCE DOCUMENTATION
STATIC, ACUTE, 48- AND 64-HOUR EC50 OP STUDY: H 19,011
TO Daphnia Hagna
AUDITS:
Items Audited Protocol Conduct Kecords, Final Report
Audit Dates 6/19/91 6/19/91 6/25/91
DATE FINDINGS REPORTED TO MANAGEMENT AND STUDY DIRECTOR; 6/25/91
Reported by:
FJ rvjj^l_^t/-/^' t~ir<'-bn fj'
Kimberyy B. Brebnei
Quality Assurance Auditor
''^l.-a. 7' Date
Company Sanitized. Does not contain TSCA Cffi
4 -
Du Font HLR 457-91
TABLE OF CONTBfrS
Page
.................................................................................................................................................................................................................................................................................................................................................................................................................................................................................. ^p
Specific Country Requirements Good Laboratory Practice Statement Quality Assurance Documentation Table of Contents
I. Summary ri. General Information
til. Purpose
IV. Records
. ....... 73.
4 5 6 7 9 9
V. Materials and Methods
10
A. Test Material
10
B. Dilution Water
10
C. Animal Culture
10
D. Test Methods
10
E. Ch'-n'ical Analysis
11
F. Statistical Analysis
11
VI. Results and Discussion
11
VII. Conclusion
11
VIII. Literature Cited
11
IX. Tables
12
Table 1. Chemical Characteristics of Haskell Laboratory Veil
Water
12
Table 2. Table 3.
Water Chemistry of the Water Control at Test Start . . J3
Physical and Chemical Characteristics of nBMHBBMHQ
Test Solutions
U
Table 4. ^Observed ImmobjJ.ity of Dap^nia magna Exposed to
.................... ^HHHHBHO01' 24, 48, and 64 Hours in a Static,
.................... 'Unaerated Test
15
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Du Font HLR 457-91
f^k------1 I. SUMMARY
L BIJ||^^^^B|vas moderately toxic to Daphnia magna neonates (less than
24 hoursoTd^i^aSunaerated, static, 48-hour test. Nominal test concentrations vere 0.31, 0.63, 1.3, 2.5, 5.0, and 10 mg/L. The 48-hour EC50 vas 8.3 with a 95Z fidiclal interval of 5.7 to 17 og/L. Due to absence of 100Z immobility in the high test concentration at 48 hours, the test vas extended to
64 hours. As a result, immobility increased in all test concentrations and the
64-hour EC50 was 0.27 mg/L.
Vork By: fi^ ^-/^A fa\^/'7/^ V. Troy Baxter/ Aoua.t ic Techni c\an
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Study
Director:
^^ //,5'Vi //-
Kevin N. Baer, Ph.D. Research Toxicologist Environmental Sciences
(dJz^l^i
Hoo//DDav/Yr
THIS REPORT IS APPROVED BOR ISSUE.
^M^ N ^gu2^
Ke^in N. Baer, Ph.D.
Study Director
^/2^/<7/
yo/^af/fr
^ Company Sanitized. Does not contain TSCA CM
Material Tested: Medical Research Number! Haskell Number: CAS Registry Number;
II. GENERAL INFOBMATLON
Du Font HLR 457-91
Other Codes:
Synonyms: Composition:
Purity; Submitier's Notebook Number:
Sponsor: Submitted By:
Study Schedule:
Notebooks:
Du Font Chemicals
W)6 E. I. du Pont de Nemours and Company
Uilmington, Delaware
Du Font Chemicals
E. I. du Pont de Neroours and Company
yilmington, Delaware 19803
Study Initiation Dates 6-17-91
Experimental Start Date: 6-18-91 Experimental Termination Date: 6-21-91 Study Completion Date: b-26-91
c^.s^.oo-.------sc.c.
Haskell Laboratory Cost Code: Distribution:
Du Font HLR 457-91
Company Sanitized. Does not contain TSCA CB1
III. PURPOSE
Du Font HLR 457-91
The goal of this study was to assess the acute toxicity of to Daphnia magna neonates less than 24 hours old.
IV. RECORDS
All data and the final report will be maintained either at Haskell
Laboratory for Toxicology and Industrial Medicii.e (Newark, Delaware) or be archived at the Du Font Records Management Center (Vilmington, Delaware).
^.
, ) Company Sanitized. Does no? confa'n TSCA C8!
A. Test Material
V. H1TKRIALS AND KETHODS
Du Font HLR 457-91
Source and composition of the test substance are described under General Information. A 10 mg/L stock solution was prepared in dilution water (see below) and used immediately. Stability and solubility of the test substance in
dilution water were not determined.
B. Dilution Water
Water (Table 1) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows, (Pimephales promelas), orior to use in this daphnid test. C- Animal Culture
Parent Daphnia magna were rearp'j at Haskell Laboratory in 2-L pyrex
beakers (25 per beaker) which contained 1.8 L of filtered dilution water held
at approximately 20C. Daphnids were fed 25 mg/L (dry weight) of a mixture of frozen Purina trout chow and Fleischmann's yeast three tines weekly during the water renewal process. Neonates were collected from approximately 29 day old parent daphnids for use in this test. Sickness, injury, and abnormality were not present. Daphnia magna were identified by labels on the beakers and test vessels.
D. Test Methods (I]
Six test concent rat ions arid a dilution water control were used. test concentrations were 0.31, 0.63, 1.3, 2.5, 5.0, and 10 mg/L.
Nominal
Pyrex beakers (250 mL;. 7-cn diameter) containing 200 mL of test solution (6.8-cm depth) were used. Each replicate (four per concentration) contained five daphnids (1 daphnid/40 mL). Daphnids were added to the control and test beakers using random numbers. Test solutions were held between 20.0 and 20.6C (mean 20.3C) and were uhaerated. Daphnids were not fed during this test. A photoperiod of 16 hours light versus 8 hours darkness was employed.
Dissolved oxygen (YSI Model 54A, Yellow Springs Instruments, Yellow
Springs, OH) and pH (Corning Model 610A, Corning Scientific Instruments,
Medfield, MA) were measured in one replicate of the water control and all test
concentrations before (*u.phnids were added at the beginning, and at the end of
the test (64 hours). Total alkalinity (Hach kit, Rach 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 before daphnids were added at the beginning of the test. Temperature "as measured using a registered American Society of Testing Materials mercury stem thermometer.
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K. Cheaical Analysis
Du Pont HLR 457-91
Chemical analysis of the test substance vas not conducted.
P. Statistical Analysis
The 48- and 64-hour EC50 was calculated using a probit method |2],
VI. RESULTS AND DISCUSSION
A rangefinding test vas conducted using the following concentrations; 0.0 (conM-ol), 0.1, 1.0, 10, and 100 mg/L. Immobilities of 20, 0, 20, 100, and 100%, respectively, were observed.
All chemical and physical parameters in the definitive test (Tables 2 and 3) vere within expected ranges.
The observed immobility at 24, 48, and 64 hours is presented in Table 4.
Daphnids were considered immobile if they weie unable to swim within 15 seconds
after gentle prodding with a glass rod. All daphnids in the high test concentration (10 mg/L) were lying on the bottom of the test vessels at 48 hours. After testing these daphnids for immobility, 602 were considered to be
immobilized. The remaining daphnids were clearly effected but were observed to briefly swim in the water column after gentle prodding and then return to the bottom of the test vessels. All mobile daphnids in the control" and lower test
concentrations appsared normal, i.e., actively swimming in the water column.
Due to the lack of 100% immobilization in the high test concentration at 48 hours, the study was extended to 64 hours (overnight). As a result, daphnids
in all test concentrations (excluding controls) exhibited immobility ranging
from 40%.in the low test concentration (0.31 mg/L) to 95% in the high test
concentration (10 mg/L). Again, all control daphnids appeared normal. The
48-hour EC50 was 8.3 mg/L with a 95% fiducial interval of 5.7 to 17 mg/L. The 64-hour EC50 was 0.27 with an undefined lower 95% fiducial limit and 1.1 mg/L
uppct 95% fiducial limit.
VII. CONCLUSION
k^^^HBHmBBRIRj1'83 moderately toxic to Daphnia magna neonates
(less t nT.Snours old) in an unaerated, static, 48-hour test. The 48-hour F.C50 was 8.3 mg/L, based on nominal concentrations and immobility.
VIII. LITERATURE CITED
1. Organisation for Economic Co-Operation and Development (OECD). t.uideline for Testing Chemicals: 202. 4 April 1984.
2. Finney, D. J. Press. 333 p.
1971.
Probit Analysis.
3rd Ed., Cambridge University
11
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TABLE 1
Du Pont HLR 457-91
Chemical Characteristjcsof Haskell_La>>oratory Veil Water1
Parameter
Concentration
Parameter
Concentration
BOD2, mg/L
<2
Lead5, ng/L
<0.005
COD, mg/L DOC3, mg/L
13
1.5
Magnesium, mg/L MBAS/LAS, ng/L
3.5 <0.05
TOC4, mg/L
Kjeldah'l N, mg/L
<0.5
O.Z
Mercury, Bg/L
Nickel, ffig/L
<0.00020 <0.026
Ammonia N, mg/L
Turbidity, NTU
Phenolics, mg/L
<0.05
<1.0 <0.050
Nitrite6, mg/L Nitrate6, mg/L
Phosphate6, ng/L
<0.1 1.6
<0.1
Color, Co/Pt
<5
Solids
Potassium, mg/L Selenium, ng/L
1.8 <0.005
total suspended, mg/L <2
Aluminum, mg/L
<0.100
Silver, nig/L
Sodium, mg/L
<0.010 8.1
Antimony, mg/L
<0.060
Sulfate6, mg/L
4.8
Arsenic, mg/L
<0.010
Sulfide, mg/L
<0.05
Beryllium, mg/L
<0.001
Thallium, mg/L
<0.01
^
Boron5, mg/L
<0.020
Zinc5, mg/L
<0.020
Cadmium, mg/L Calcium, mg/L
<0.002 25.0
Volatile priority
pollutants
ND"
Chloride6, mg/L
Chromium, mg/L
6.3 <0.010
Acid cxtractable priority pollutants ND"
Cobalt, mg/L
<0.020
Base/neutral
Copper, mg/L Cyanide, mg/L
<0.0in <0.02L
priority pollutants ND
Pesticides/PCBs
ND8
Iron6 mg/L
0.053
Organophosphate
Fluoride6, mg/L
<0.1
pesticides7, ug/L
<0.50
lDate of analyses 17 October 1990 unless indicated otherwise, analyses
performed at Environmental Testing and Certification Corporation, Edison, New Jersey; ^ate of analyses 13 December 1990, analyses analysis performed at Environmental testing and Certification Corporation, Edison, New Jersey; 'Muan value; "Date of analyses 24 July 1990, performed at Du Pont Engineering Test Center, Newark, Delaware; ^eloy the Published Method Detection Limit (BMDL);
'Date of analyses 9 November 1990, analyses performed at Du Pont Chemicals,
Jackson Laboratory, Deepwater, New Jersey; ^ate of analyses 23 October 1990, analyses performed a'' Hazleton Laboratories, Inc., Madison, VI; 'None detected.
I) 12
Company s^.-."-"0'""*""TM"81
Du Font HLR 457-91
TABLE 2
Water Chemistry pf the Water Control at Test Start
Total Alkalinity
(mg/L as CaC03)
82
EDTA Hardness (mg/L as CaCOy)
75
Conductivity
(limhos/cn)
170
13
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^
,,
^
^
^
^
TABLE 3
Du Font HLR 457-91
Physical and Cheatcaj. Characteristics of Fest Solutions
Nominal Concentrations (og/L)
Timfc
(Hours) Water Control 0.31 0.63 1.3 2.5 5.0 10
Dissolved Oxygen (mg/L)
0
8.8
8.7
8.7
8.7 8.7 8.7 8.7
64
8.7
8.6
8.6
8.6 8.5 8.4 8.4
pH
0
7.6
7.6
7.6
7.7 7.7 7.7 7.7
64
7.6
7.7
7.8
7.9 8.0 8.0 8.0
14
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TABLE 4
Observed :Immobiility
for 2>^, 48,> and
WOn ~ of Daplrinia magna
64JHo,ii
^ ^ E?cposed toHB
Lc, Un.ie rated
Per(:ent Cuinulativ e Obs<irved :[nmobil ity
A
Nominal
Concentrationsi (g/L)
24 1Bours
A
B*
C*
D*
48 lours A- B- C* D"
64 1Hours A* B* C" D"
Water Control
0
0
0
0
0
0
0
0
0
0
0
0
0.31
0
0
0
0
0
0
0
0
20 60 60 20
*
i
0.63
0
0
0
0
0
0 20
0
100 60 60 80
1.3
0
0
3
2.5
VI
i\i
^
5.0
1
10
0
0
0
0
0
0 20
0
0 20 20 20
20 20 40 40
0
0
0
0
0 20 20 20 20 40 20 40 401' 60t> 60" 80"
80'' 60 100 60" 60*' 100 100 100 20i 60 100 100 80t> 100 100 100
N"
Rl
a.
0
A, B, C and D represent replicate test vessels containing five daphnids each.
0
1)
i(^
3
"All "mobile'' daphnids vere lying on the bottom of the test vessels and vere obi Ie on
'-I
after gentle prodding with a glass rod (see Discussi(in for further cooaents)
3
:
i
-1 a
0 5
0
S3
w