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Study TitU
STATIC ACUTE 48-HOUR EC50 OF
DAPHNIA MAGNA
Data Requirgment
United State? Environmentfti Protection Agency Bnvironaontal Effects Test Guidelines Subdivision EG-1
Author Kevin N. Baer, Ph.D.
Study Completed On
16 July 1990
Haskell
Performing Laboratory
i
E. I. du Pont de Nemours and Company, Inc.
Laboratory ior Toxicology and Industrial Medicine Elkton Road. Post Office Box 50
Newark, Delaware 19714
Laboratory Project ID
i
Haskell Laboratory Report No. 349-90
Page 1 of 13
onspany Sanitized. Does not contain TSCA CB8
Du Font HLR 349-90
'ATBffiNT
This study vas conducted
to U)(iId^St|i^es Environmental Protection
Agency's Good Laboratory
Regulati^fr <40 CFR 792) for TSCA. Areas of
non-coapllance are docuoentejd in the^tudyirecords. No deviations existed
that significantly affected |the validity of^fbe ^tudy.
Sponsor: Submitter:
E. I . du Font de Honours and Co., xnc. Chein|icals and PigBents Department
Study
Director:
/^^k /U. ^LtiLA
Kevtii N Baer, Ph.D. Rese arch Toxicologlst Environmental Toxicology and
Information Services
S.'J^e.^O
Day/Mo/Yr
Company SanRized. Does not contain TSCA CBI
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Du Font BLR 349-90
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QUAIJ^YASSUB^g^E9(^EATION
-UDY;
H 18,267
STATIC ACUTE 48-HOUR EC50 OF|
AUDITS: Items Audited
Protocol, Conduct Records, Report
Audit Dates 5/24/90
7/5-6/90
FTO DAPHNIA HAGNA
J ^ l SHORT-TERM AUDIT REPORT NUMBER:
DATE FINDINGS REPORTED TO MANAGEMENT AND
STUDY
DIRECTOR:-7/6/90
[
Reported by: l^^--^/ <s^^--
j Villlam J. Lynam
Quality Assurance Auditor
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Date
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Du Font BLR 349-90
Page
Good Laboratory Practice St imnent
2
................. Quality Assurance Docunenta :lon
3
................... Table of Contents
.j
4
..................... .... I. Purpose
5
...... ...... .1.............. II. RInetcroorddusction/Summary 55
.........I;....................................... General Information ..!
6
..................... III. Materials and Methods
7
A. Test Material
7
.!..................... B. Dilution Water
7
........................ C. Animal Culture
7
....................... D. Test Methods
7
....................... E. Chemical Analysis
8
............................................. IV. Results and Discussion
8
'
..................... V. Conclusion
8
........................... VI. Literature Cited
8
.....l.f..|...|................................................................................. .... fB9 ......... VII.
Tables Table 1.
Table 2. Table 3. Table 4. Table 5.
\.
Chemical Characteristics of Haskell Laboratory Well
Water
1.
Water Chemistry of the Water Control at Test Start .
Dissolved Oxygen Concentration of
pH of
Observedlmmo^ility of Daphnia magna Exposed to
for 48 Hours in a Static, Unaerated Test
9
9 . 10
11 12
13
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Du Font HLR 349-90
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The objective vas to Wf^an the (icuiMs'^-^^etty
aagna
neonates
less
than
24
..
hours
old."
: ";
of
fl9
to
Daphnia
II. iiiiitopocrfon/;
_____fexhibited ho acute toxleity t^ Pa^inia mapna neonates (less than
24 hours old) In an unaeratrd, static 48 hour ^st. Nominal concentrations
were 1311, 1638, 2048, 2560 3206, and 5000Bg/L. The 48-hour EC50 could not be calculated because 5000 iig/L, the highest concentration tested, failed to
cause immobilization of the daphnids.
!
BEOOBDS
All data and the finaljreport will be maintained either at Haskell Laboratory for Toxicology and Industrial Medicine (Newark, Delaware) or be
archived at the Du Font Records Management Center (Vllnington, Delaware).
^ L ^ / L ^ Technician: Charles :-. Hall Environmrntal Toxicology and
Informat -on Services
T^r ^ 7
9o
Day/Mo/Year
M^^/u' ^L^ Study Director: _/[W^ /U' Kevin N.IBaer, Ph.D. Researchj Toxicologiat
Snvis-onintental Toxicology and Information Services
^7 S)^c ^0
Day/Mo/Year
THIS REPORT IS APPROVED ^OR ISSUE.
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III.! MATERIALS AND METHODS Du Font HLR 349-90
A. Test Material
Source, composition and! purity ofH^^^^are described under General
Information. The anpropriate quantity
for each concentration was
ot^Up weighed and dissolved in 1 liter of filtered fish tank water (see below) and
and then distributed into four 0.25 liter containers in the water bath.
Stability and solubility
in dilution water were not determined.
offBI '
B. Dilution Vbter
Water (Table 1) originated from the Baskell Laboratory well and flowed through aquaria containing fathead minnows (Pimephales promelas) prior to use in these daphnid tests.
C. t^sol Culture
Parent Daphnia magna were reared at Baskell Laboratory in 2-L pyrex beakers (21 per beaker) which contained 1.8 L of filtered dilution vater held at approximately 20C. Daphnids were fed 30 ng/L (dry weight) of a mixture of frozen Purina (same as Buckshire) trout chow and Fleischnann's yeast three times weekly during ttie water renewal process. Neonates were collected from 15 day old parent daphnids for use in these tests. Sickness, injury or abnormality was not present. Animals were identified by labels on the beakers
or test vessels.
D. Test Methods [1]
Six test concentrations and a water control were prepared using filtered fish tank water. Nominal test concentrations were 1311, 1638, 2048, 2560,
3200 and 5000 mg/L.
Pyrex beakers (250 roL; 6.1-cm diameter) containing 200 mL of test solution (6.8-cm depth) were used. Each replicate (four per concentration)
contained five daphnids (<0.1 g daphnld/L). Daphnids were added to the water control and to test beakers beginning with the lowest concentration. Random
assignment was not used to avoid contamination of test solutions of low concentration from the transfer pipette when daphnids were released under the
water surface. Daphnids were not fed during the test. Test solutions were held between 19.9 and 20.1C (mean IQ.O^C) and were unaerated. A photoperiod of 16 hours light (506 to 592 Lux) versus 8 hours darkness was employed.
,,.,.-------
W?SSSt^S:y^
^^?y| Dissolved oxygen
replicates of every f^il^c8"^
beginning and at the ^emt^f 'jfl conductivity of the dilution?!! Tenperatures were taken yith'-i calibrated mercury therapDeterj
B. Chexical analysis
No chemical analyses of
Ou Font HLR 349-90
iter controls and in all
lids vere added at the
iity, EDTA hardness and the beginning of tlie teat*
Testing Materials
test solutions yejce perforBed.
. RESULTS AND DJSCVSSION
An earlier rangefinding
reported in this study.
vas conducted but the results will not be
TT.ns solutions of fl^----1exhibited a Bllky white color. All chemical
and physical parameters (Tables 2, 3 and 4) were within expected ranges. The 48-Hr EC50 could not be calculated because 5000 mg/L, the highest concentration tested, failed to cause iBBiobilizatlon of the daphnia (Table 5).
V. COSCLUSIOH
'exhibited no acute toxiclty in this static, unaerated 48-hour Daphnfa magna test. The EC50 u&s greater than 5000 og/L, the highest
concentration tested.
VI. LITERATURE CITED
1.
United States EnvlrpnBental Protection Agency, Office of Toxic
Substances, Guidelines for Tes (ting Chemicals, EG-1, Washington, DC., August
1982.
Company Sanitized. Does not contain TSCA CBi
Du Font HLR 349-90
TABLE 1
Chemical Character! jstics of Bsskell Laboratory Veil Vateri
Paraaeter
Concentration
Paraaeter
Concentration
BOD, mg/L
COD, mg/L
DOC, og/L
TOC2, Bg/L Kjeldahl N, mg/L Aomonia N, og/L Organic N, mg/L T--bldifcy, MTU
Phenolics, mg/L Color, Co/Pt Solids
total suspended,
Aluminum, mg/L Antimony, mg/L Arsenic, mg/L
Beryllium, mg/L
Boron, mg/L Cadmium, mg/L
Calcium, og/L .Chloride3, mg/L Chromium, mg/L
Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron3, mg/L
mg/L
5
<10
<lr0 0.8 <0.2
<0j,2
<0,2 <lg.O <0.05
<5
<4
<0,100 <0,060 <0.010 <0.001 <0,021 <0p002 25,4 6.3 <0.010 <0.020 <0,010 <0.025 0.131
Pluorlde', mg/L
Lead, mg/L Magnesium, &. 'L MBAS/LAS, mg/L
Mercury, mg/L Hickel, mg/L
Nitrite3, mg/L Nitrate3, rog/L
Phosphate3, fflg/L Potassium, fflg/L
Selenium, mg/L
Silver, ng/L
Sodium, mg/L
Sulfate3, mg/L Sulfide, mg/L Zinc, mg/L
Volatile priority pollutants Acid extractuble priority pollutants
Base/neutral
priority pollutants
Pestlcides/PCBs
Organophospha t e
pesticides^, ug/L
<0.l
<0.075 3.9
<0.05 <0.00020 <0.020 <0.1
1.7 <0.1
1.8 <0.005 <0.010 8.1 5.0 0.11 <0.020
MDS
HD6
ND6 MD<
<0.50
lDate of analyses 24 October 1989 unless Indicated otherwise. 2 Date of analyses 5 December 1989. 3 Date of analyses 11 December 1989. ^Date of analyses 26 September 1989. SNone detected except methylene chloride vas belov the published Method Detection Limit (BMDL).
None detected.
noic^8'"^5'CP^
>niied. Does
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TABLt ;?:
Water Chenist]ry of the Water Control at Test Start
Total Alkalinity
(ng/L as CaCO,)
EDTA flardness* (Bg/fc as CaCOia)
COt.ductivity1' (Vinhos/cm)
76
77
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Concentration (Bg/L)
0 Hours
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8.8 8.8 8.8 8.8
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8.8 8.8 8.8 8.8
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S.8 8.8 8.8 8.8
8.8 8.8 8.8 8.8
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2 Five daphnids in each rjeplicate.
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7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
lall
1638 2048
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
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2560
--/
3200
i
7.7
7.7 7.7
7.7 7.7 7.7- 7.7
7.7 7.7 7.7 7.7 7.7 7.7 7.7 7.7
5000
7.7 7.7 7.7 7.7
7.7 7.7 7.7 7.7
corning Model 610A, C<irning Scientific Instruments, Medfield, HA. 2 Each replicate contali ied 5 daphnids.
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^ercent Cumulative Observed Immobility
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48 Hours
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Nominal
r.
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Concentration (mg/L)
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0
0 0 0 1311
0
0
0
0000 OjO '
0 0 0 ' 1638
0000 OJOOO f
2048
:
0!000 0000 1 2560
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o!o00 0000 ;
3200
0000 0000 j i]: i
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5000
0,000 0000
1 *
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^ach replicate contairi ed five daphnids.
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