Document X76nwyVM1qvGJzn4xdba5xbvw
AR226-2976
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Study Trtle
STATIC. ACUTE. -<-HOyn^ OF
H-19,520 TO Dapfyniamagna naj^BsauiremfiDi
Organisation for Economic Co-Operation and Development (OECD) Guideline for Testing Chemicals: 202
Author Keith B. Piecsori. Ph.D. Study Completed On
July 10,1992
Performing Laboratory E. 1. du Port de Nemours and Company Haskeit Laboratory for Toxicology and Industrial Medicine
Elkton Road, Post Office Box 50 Newark, Delaware 19714
Laboratory Project ID Haskell Laboratory Report No. 437-92
Page 1 of 19
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THIS PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREMENTS
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E. ftemi^i^^is............................^.;...-...............,........................................^ R
Sia8^^XN^s..........................^,,.........^........,,..........................................19
IV. Results and Discussion .......................................-...;...,...........................................l 0
V. Conclusjon...................................-.............................,,....,................"^ VL Uterature Cited...........................................................,..................................................11
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1. Cfes.-nca{ Cbar-sefteiisii'es @fNas^ii
^^ei! Water.......................... 12
2. Water Chemistey of the 01(utj.n ^atr at ti&sB^lift.................^......................13
3. Physical fi^jl^iejaii^CAajaaeal^^
Test Solutions.............! 4
4. Observed lnlro6Nityof0^fiw^^i|!^?|^N8;to N-19,52 for
48 faouts in a^tatfc, UnaeBateitfTi^....^....^..,^........................................l6
5. H-IS.NO heentrattons Cau?iragIrnBisblfi^t 0% (EC50)
of Daphnia magnass. SoedficTimS lr^ewals.................................--................l 7
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Study Director - 6/25/92 Management 7/W/92
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Deborah A. \^fck Quafity AssLirance Auditor
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II. RECORDS
to Qaphnia magna
All data and the final report will be maintained either at Haskel! Laboratory for Toxicology and industrial Medicine (Newark, Delaware} or be archived at the Du Pont
Records Management Center (Wilmington, Delaware).
Hi. MATERIALS AND METHODS
A. Test Substance
The test substance is described under General Information. A stock solution (1100 mL) was prepared by weighing the approp iate quantity of hM 9,520, adding it to dilution water (see below) and stirring for 30 minutes. Individual test solutions (262 mL per flask) were then prepared from this stock in dilution water. Each of the above solutions was kept in a sealed container with minimal head space to avoid loss of the test substance through volatility. The stability and solubility of H-19,520 in dilution water (see below) and under storage conditions were not determined.
B. Dilution Water
Water (Table 1.) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows, Pimephales promelas, prior to use in this daphnfd test.
C. Animal Culture
Parent Daphnia magna were reared at Haskell Laboratory in 250 mL Pyrex beakers (1 per beaker) which contained 200 mL of filtered dilution water or autociaved fi.'sred dilution water held at approximately 20C. Daphnids were fed two alga! species, Ahkistrodesmug faiestos and Selgnsstrum capricomutum. at a rate of 75,000 cells/mL of each- spec-fes three times weekly daring the water renewal process. Neonates (less than
24 hours old) were collected from '14 day old parent daphnids for use In this test.
Sickness, injury, and abnormality were not observed. Daphnia magna were identified by iabeis on the beakers and test vessels.
D. Test Methods [1]
Eioht test concentrations and a water control were used. Nominal test concentrations
were 7.82,15.6, 31.3, 62.5,125.251. 500 and 1000 mg/L.
9 Sompany Sanitized. Does not contain TSCA CBI
would nm flttftteugfttN aecit FHiitSrtlBiN^irl
Chemical Company, tj@el^til, CD|,Sl0mfe%Agf|^^Hgpn(at, Hach Gftera Company, LoveSand, 0). and laaBdaiStiw^P^isSSa1^. Yeliow Springs
instruments, Yeilow Springs, OH) of the ditutson water were measured before
daphnids were a^wS at She beginning oftteeie^t. X Q^nltinuously-recording
thermometer was used! in a dummy conttisi 1checiklpr iemperature variatton during
the 48^our test. E. Chemical Analysis
Concentrations of H-19,520 in test solutians were not measured.
F. Statistical Analysis The 24- and 48-hour E050s and their associated 95% IRducial limits were caJeuiated by the probit method using the logarithm of the concentration [2j.
IV. RESULTS AND DfSCUSSSON A 48-hour rangefinding study was conducted in uncovered test chambers. Test concentrations were 0.1,1,10,100, and 1,000 mg/J- ahtfa water control; observed
immobilities were . 0.0.0,100, and 0%. irespactively. ^SobHedaphnids in the
100 mg^. concentration appeared sluggish. The definitive study was conducted using nominal concentrations of 7.82,15.6. 31.3, 62.5.125, 251,500, and 1,000 mg/L The test chambers were seated to prevent toss
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VI. LITERATURE CITED
1. TOersgtainnigsa^tEionnjf^erfiecf;Miom20ic2.Co4-AGppr^irla11ti9e8n4.and Devejepment (OECD). Quidetine for 2. SThAeS PInrsotNfttuPte nInie^.,w<^ea.ry,1N9C85. . SAS* IJse^sQuNe1: ^atistte. Version 5 Eclitton.
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ChsmscssJ Characteristics of Haskel! Laboratory Well Water1
Parameter
Concentration
Parameter
Concentration
BOD, mg/L COB. mQfL DOC. mg/L TOC2 mg/L
Total Kjeldahi N, mg/L Ammonia N. mg/L
TufN^ly. NTU Phenelics, mg/L
Color, Co/Pt
Soiids TotaS suspended, mg/L Aluminum, mg/L Antimony, mg/L
Arsenic, mg/L Beryllium, mg/L
Cadmium, mg/L
Calcium, mg/L Chloride3' mg/L Chromium, mg/L Cobalt, mg/L
Copper, mg/L Cyanide. mg/L srehS, mg/L
Ruoride3
<2 <10
2.8 <0.5 <0.2
<O.Q5
1.6 <Q.Q50 <5
<4 <0.100 <Q.060 <0.010 <0.001 <0.002 24.5 6.2 <0.010 <0.020 <0.010
<Q,Q25
0.025
<0.1
Lead, ni@/L Magnesium, mg/L MBAS/LAS, mg/L Mercury, mg/L Nickel. mg/L Nitrite3, mg/L & *. . * ---'
PRbsphat^3, mg/L
Potassium, mg/L Selenium, mg/L Stiver, mg/L Sodium, mg/L SuEfate3, Rig/L
u(fide, m5/L ThaHium, mg/L
2nc4, mg/L Volatile priority
pollutants Acid extractable priority poHutants
Base/neutral priority ppHutants
Pestieides/PCBs Organpphosphate pesticides5, ng/L
<0.075 3.8
<0.05 <0.00020 <0.020 <0.1
2.1 <0.1
1.8 <0.005 <0.010
8.0 4.8 <0.05 <0.01 <0.02Q
NDS
NDS
ND ND6
<0.50
^Date of sample collection 30 December 1991 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; ^Date of sample collection 15 January 1992, performed at Du Pont Engineering Test Center,
Newark, Delaware; ^ate of analyses 13 January 1992, analyses performed at Pu Pont
Chemicals, Jackson Laboratory, Deepwater, New Jersey; -^Below the Published Method Detection Limit (BMDL); analyses performed at Hazleton Laboratories, Inc., Madison, Wl;
^one detected.
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Conductivity (nmhosfcm)
180
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5tart
Nominai
Concentrations
N/L)
^r
B8
C
a
48 Hours
Aa
Ba
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HgO Control
8.8
b
7.82
8.6
b
15.6
8.5
b
31.3
8.6
b
62.5
8.6
b
125
8.6
b
251
8.6
b
500
8.6
b
1000
8.7
b
6H
HaO Control
7.3
7.82
7.3
' >
15.6
7.3
l i
31.3
7.3
* >
62.5
7.3
t '
125
7.4
t
251
7.4
b
500
7'4
b
1000
7.4
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
fe
b
1
b
1
b
b
b
tr
b
i i
b
fc
b
b
8.5
8.6
8.5
8.5
8.6
8.6
8.5
8.6
8.6
8.5
8.4
8.4
8.4
8.5
81.6
8.5
8.5
8.5
8.5
6M
8.5
8.5
8.4
8.5
8.6
8.5
8.5
8.5
8.5
8.4
8.4
8.5
8.4
8.4
8.5
8.4
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7,5
7.5
7.5
7.5
7.S
7.5
7.5
7.5
7.5
7.5
7.5
7^
7.S
7.5
7.5
7.S
7.S
7.5
7.5
7.5
7.S
7.5
7.5
7.5
7.5
7.5
7.5
7.5
7.5
"A, B, C, and D represent reph'eate test vessets containing five daphnicfs each at te$t start.
'See last paragraph of D. Test Methods for explanation as to why a single reading was made.
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TelWatyre.a)C
HgO Control
20.2
b
b
b
7.82
20.2
b
b
b
15.6
20.2
b
b
b
31.3
20.2
b
b
b
62.5
20.2
b
b
b
125
20.1
b
b
b
251 500
20.2
b
b
b
20.2
b
b
b
1000
20.2
b
b
b
19.9 19.9 19.8 19.8 19.9 19,8 19.9 19.9 19.9
19.9 19.8 19.8 19.8 19.9 19.8 19.9 19.9 19.9
19.9 19.8 19.8 19.9 19.8 19.8 19.9 19.9 19.9
19.9 19.8 19.8
19.9
is.ais.e
19.9 19.9
19.9
^ B, C, and D represent repiicate test vessels containing five daphnids eac:- attest
start.
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'See last paragraph of D. Test Methods for @xp!ar8gtJE?ans to why a sing! reading was
made.
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Water Contro!
0
0
0
0
7.82
0
0
0
0
15.@
0 0 0 Q. -
31.3
62.5 125 251
0
0
0
0
0
0
0
0
0
0
0
0
40
20
0
0
500
20
40
40
20
1000
SO
40
20
60
0
0
0
0
0
0
0
0
0
0
0
0
0
0
20
40
40
20
40
0
40
60
40
BO
80
40
60
80
80
60
80
100
80
80
60
"A, B, C and P represent replicate test vessels containing five daphnids each at test start
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Qa Font %LR 437-92
PROTOCOL AMENDMENT NUMBER ONE
|
a. Tiie deflnition of jmmobHjzation on page 8 of SOF'lplBHBH^
In lieu thereof immobilization wil! b9 defined as the inlbJIJtyto swim {at least two body
Sgngths in any one direction) within 15 seconels after ^entfe prodding with a giass rod.
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b. Daphnids will be distributed to test chambers using random numbers. No more
;
than two daphnids will be placed into one chamber per random number.
c. A registered (not calibrated) American Society of Testing Materials mercury
;
thermometer will be used to take temperatures of test solutions.
'
d. Analytical measurements of test solution concentrations will not be made.
Q. Surviving daphnids at test end wilt be sacrificed according to SOFJIHHI
t Soilubdityand stability studies of the test substance in dilution water will not b performed.
''-^l}-! g. inMHIHHHHHEp" ^acse four
thann l1o0%%^^Tftnhe daphnids diiee or show
tne sentence "a test is no! acceptable if abnormal behavior in either control" is
more
ambiguous. The sentence should read "a test is not acceptable if more than 10% of
the daphnids die or show abnormal behavior in any of the four replicate controls".
h. Beakers being acclimated prior to use in the definitive test will be covered by plastic-coated paper (plastic side down)-not glass as described in the SOP.
1. Temperatures will be taken at the start of the test before addition of daphnids and at the end of the test after the final immobilization readings are made. Temperature variation will be assessed by a continuously-recording probe which will be placed in a dummy control vessel sitting adjacent to one of the four water controls in the water
bath.
PROTOCOL AMENDMENT NUMBER TWO
a. Based on the initial Range-finding test, the concentration levels (Test Substance) wilt be:7.82,15.6, 31.3, 62.5,125. 251, 500,1000 mg/L to increase the probability that zero immobilities will occur at the lowest test concentration.
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6. ^me that vi^ter bath temperature was taRen was nol:doeumented in study records.
c. Pre'-^RdttlQii^g of t@gt vessels wsis net
l|;3n;Situdrsycords.
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