Document jNVnDx3XpK1VvZpXQZzp3KjDZ
AR226-2975
TRACE SECRET Study Title
STATIC, ACUTE, 48-HOUR EC50 OF H-19,519 TO Daphnia magria SafoBgauireroent
Organisation for Economic Co-Operation and Development (OECD) Guideline for Testing Chemicals: 202
Author Keith B. Pierson, Ph.D. Study Completed On
July 1, 1992 Performing Laboratory E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, Post Office Box 50 Newark, Delaware 19714
Laboratory Project ID Haskell Laboratory Report No. 460-92
Page I o f 16
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THIS PAGE RESERVED FOR SPECIFIC COUNTRY REQUIREUPMTS
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Do Pont HLR 460-92
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to EPA FIFRA and TSCA Good Laboratory Practice
Standards (40 C FR 160 and 40 CFR 792), OECD Principles o f Good Laboratory
rac*' ' (C(81)30(Fina!), Annex 2), and MAFF Japan Good Laboratory P rance
Star.
(59 NohSan No. 3850). Solubility and stability of the test substance in
dilution water and under storage conditions were not determined. Concentrations of the
tost substance in dilution water were not measured. 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.
Submitter: E. I. du Pont de Nemours and Company
Sponsor: STUDY DIRECTOR:
Du Pont Chemicals E. I. du Pont de Nemo, rs and Company
Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences
Z3. ftm Z iW X Mo/Day/Year
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Do Pont H LR 460-92
TABLE OF CONTENTS SpedSc Country Requiremens
Page ---------- 2
Good Lsboratory Practica Statement... laoie o?uontents.__ Genera! Information..
....------.3 ...........A .............5
Summary.
'ft
Quality Assurance Documentation........... L Purpose_____ IL Records.----------
.....7 ..............8 -----------8
Ilf. Materials and Methods ....
A. Test Substance......... B. Dilution W ater............
t e t t s t t a u <
0. Animal Culture............
D. Test Methods.............
3 E. Chemical Analysis..... IV. Results and Discussion ....
------- 8
.............8 8 ................
.8c o c a s c . e . a
.......... 9
-------- 9
V. Conclusion.......................
* s e s e e :,,,....,,,10
Vi. Literature Cited...................
........ 10
VII. Tables...................................................................................................... ....... 11
1. Chemical Characteristics of Haskell Laboratory Weil Water.. 2. Water Chemistry of the Dilution Water at Test Start.....
........ ...............1112
3- Physical and Chemical Characteristics of H-19,519 Test Solutions..." ...... 13 4. Observed Immobility of Daphnia magna Exposed to H-19,519 for
48 hours in Static, Unaeraied Test.......................... ..... ..................... ......15
VIII. Appendices ........................................................................................ .-- 16 i. Protocol Amendments and SOP Deviations................. ....................... ..... 16
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Du Pont HLR 460-92
Sponsor: Blugi/Joifiated/Com pfeted:
Du Pon Chemicals E. i. du Pont de Nemours and Company Wilmington, Delaware 19898
6-8-92 / 7-1-92
6-10-92/6-12-92
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Du Pont H LR 460-92
STATIC, ACUTE, 48-HOUR EC50 OF H-19,519 TO Daphnia magna
SUMMARY
H-19,519 was not toxic to Daphnia magna neonates (Jess than 24 hours old) in an unaerated, static, 48-hour te s t. Nominal concentrations were 15.6, 31.3,62.5,125, 250,500 and 1000 mg/L. The 48-hour EC50 could not be calculated because no immobilities occurred at the highest concentration tested.
Work By: Robert W. Peterson, B.A. Aquatic Associate
JeffreyT. Turner Aquatic Assodate
C/s.2, h a Mo/Day/Year
THIS REPORT IS APPROVED FOR ISSUE.
'X L $ . P ttlS aJ
Keith B. Pierson, Ph.D. Research Toxicologist and Study Director Environmental Sciences
/ Tt.y / f fa . Mo/Day/Year
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Du Pont HLR 460-92
QUALITY ASSURANCE DOCUMENTATION (H-19519)
Oates of inspection:
C o n d u ct-6 /1 0 /9 2 Records, Report(s) - 6/24,25/92
Findings reported to:
Study Director - 6/29/92 Management - 7/1/92
Reported by:
th n U x A I jJ A - fy 'lJ jrL ts ' Kimberly B. Ewebner Quality Assurance Auditor
n h l9 ^ Date
w-
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I. PURPOSE
The goal of this study was to assess the acute toxicity of H-19,519 to Daphnia magna neonates less than 24 hours old.
II. RECORDS
AH rfata and the final report m il be maintained either at Haskell Laboratory for Toxicology and Industrial Medicine (Newark, Delaware) or be archived at the Du Pont Records Management Center (Wilmington, Delaware).
III. MATERIALS AND METHODS
A . Test Substance
The test substance is described under Genera! information. Each 1100 mL solution was prepared by weighing the appropriate quantity of H-19,519, adding it to cfilution water (see below) and stirring for 60 minutes. Ec.ch solution was allowed to come to equilibrium test temperature (see below). The solution was then apportioned among the four replicate test chambers (262 mL per beaker) using a graduated cylinder. Each of the above solutions was kept in a sealed container with minimal head space to avoid loss o f the test substance through volatilization, i ne stability and solubility of H-19,519 in dilution water (see below) and water storage conditions were not determ ined.
B . Dilution W ater
W ater (Table 1) originated from the Haskell Laboratory well and flowed through aquaria containing fathead minnows, Pimephales promelas, prior to use in this daphnid test.
C . Anim al 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 autoclaved filtered diiytion water held at approximately 2QC. Daphnids were fed two alga! species, Ankistrodesmus falcatus and Selenastrum capricomutum. at a rate of 75,000 cells/mL of each species three times weekly during the water renewal process. Neonates (less than 24 hours old) were collected from 19-35 day old parent daphnids for use in this test. Sickness, injury, and abnormality were not observed. Daphnia magna were identified by labels on the beakers and test vessels.
D . Test Methods [1]
Seven test concentrations and a water control were used. Nominal, test concentrations were 15.6, 31.3, 62.5,125, 250, 500 and 1000 mg/L.
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Du Pont HLR 460-92
Screw-top Erienmeyer flasks (262 mL; 8-cm maximum diameter; 12.5-cm solution depth; and 13-cm total height) closed with a valve assembly (Miniert) was used. Each flask contained ^proxim ateiy 262 mL of test solution and was filled to the top to minimize head spat and thus lim it loss of test substance from volatility. Four replicates were used to r each test concentration; each replicate contained five daphnids (1 daphnid/40 mL) fo r a total o f 20 daphnids per test concentration. Daphnids were added to the control and test beakers using random numbers. Test solutions were held between 20.1 and 20.5C (mean 20.3C) and were unaerated. Daphnids were not fed during the te s t. A photoperiod o f 16 hours light (538 to 592 Lux) versus 8 hours darkness was employed with approximately 25 minutes of transitional light (10.8 to 16.1 Lux) preceding and following tee 16 hour light interval. Observations were made daily.
Dissolved[ oxygen. (YSiMode! 54A, Yellow Springs Instrum ent, Yellow Springs, OH), pH (Coming Model 61OA, Coming Scientific instrum ent, Medfieid, MA), and temperature (registered American Society of Testing Materials mercury thermometer) were measured at test start in each 1100 mL test solution and the water control after they reached test temperature equilibrium, an d. i ali replicates of the water control and all test concentrations at the end of the test. Measurements of individual replicate concentrations could not be made without loss of potentially volatile test substance components and because the pH and dissolved oxygen probes wouid not fit through the neck of the Erienmeyer flasks. 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 before daphnids were added at the beginning of the test. A continuously-recording thermometer was used in a dummy control to check for temperature variation during the 48-hour test.
E. C hem ical A n a lysis
Concentrations of H-19,519 in test solutions were not measured.
IV. RESULTS AND DISCUSSION
k 48-hour rar.gefincfing study was conducted in sealed test chambers. Test concentrations were 1,10,100, and 1,000 mg/L and a water control; observed immobilities were 0 ,0 , 0 ,0 , and 0%, respectively. Undissolved test substance In all test concentrations suggested that the test substance was not entirely in solution.
The definitive study was conducted using nominal concentrations of 15.6, 31.3, 62.5, 125,250, 500, and 1,000 mg/L. The test chambers were sealed to prevent loss of potentially volatile elements of the test substance. Again undissolved test substance and surface films in ail test concentrations suggested that the test substance was not entirely in solution. All chemical and physical parameters (Tables 2 and 3) were within expected ranges.
Immobilization was defined as the inability to swim at least two body lengths in any one direction within 15 seconds after gentle prodding with a glass rod. No immobilities
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S S S t o t f e m iS "*toCateula8dbecause",mmoM,ies
V. CO NCLUSION
^SSSiStSSSSST^ma9naneonates(|9SS" 24hou,s*
VI. LITERATURE CITED
1. Organisation for Economie Co-Operation and Development (OECD). Guideline for
Tesirng Chemicals: 202. 4 April 1984.
6 ror
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Du Pont HLR 460-92
STATIC, ACUTE.48-HOUR EC50 OF H-19,519 TO D aphnia m agna
TABLE 1
Chemical Characteristics of Haskeli Laboratory Weil Water1
Parameter
Concentration
Parameter
Concentration
BOD, mg/L
DOC, mg/L TOC2, mg/L Total Kjeidah! N. mg/L Ammonia N, mg/L Turbidity, NTU Phenolic, mg/L Color, Co/Pt S o lid s Total suspended, mg/L Aluminum, mg/L Antimony, mg/L Arsenic, mg/L Beiyljium, mg/L Cadmium, mg/L Calcium, mg/L Chloride3- mg/L Chromium, mg/L C obalt mg/L Copper, mg/L Cyanide, mg/L Iron3, mg/L Fluoride3
<2 <10
2.8 <0.5 <0.2 <0.05 1.6 <0.050 <5
<4 <0.100 <0.060 <0.010 <0.001 <0.002 24.5 6.2 <0.010 <0.020 <0.010 <0.025 0.025 <0.1
Lead, mg/L Maanesium. ma/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 Thallium, mg/L Zinc4, mg/L Volatile priority pollutants Acid extractable priority pollutants Base/neutral
priority pollutants P e sticid ts/P C B s OrganopKosphate pesticides5, gg/L
<0.075 3,3
<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 <0.020
ND5
ND
ND ND
<0.50
1Daie of sample collection 30 December 1991 unless indicated otherwise, analyses
performed at Environmental Testing and Certification Corporation, Edison, New Jersey;
2Date of sample collection 15 January 1992, performed at Du Pont Engineering Test Center Newark, Delaware; 3Date of analyses 13 January 1992, analyses performed at Du Pont ' Chemicals, Jackson Laboratory, Deepwater, New Jersey; 4Be!ow the Published Method
Detection Limit (BMDL); 5Analyses performed at Hazleton Laboratories, ine., Madison, Wl; None detected.
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STATIC* ACUTEi 48-HOUR MG50 OF H-19,S9 TO Daphnia m $ii
TABLE 2 Water Chemistry of the Dilution Water at Test Start
Total Alkalinity {mg/L as CaCOa)
85
EDTA Hardness (mg/L as CaCOs)
85
Conductivity (pmhos/em)
180
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STATIC, ACUTE. 48-HOUR EC50 OF H-19,519 TO D aphnia m agna TABLE 3
Physical and Chemical Characteristics of H-18,519 Test Solutions
-- Mamina!-....
Concentrations (mg/L)
Aa
Start ga Ca
D
48 Hours Aa B Ca Da
Dissolved Oxygen. mo/L
HgO Control
15.6 31.3 62.5 125 250 500
1000
8.1
8.1 8.0 8.1 8.1 8.0 8.0 8.0
b b b b
b b b b
H2O Control 7.2 b
15.6 7.2 b
31.3
7.3 b
62.5
7.3 b
125 7.3 b
250 7.3 b
500 7.3 b
1000
7.3 b
bb bb bb bb bb bb bb bb
bb bb bb bb bb bb bb bb
8.1 8.1 8.2 8.1 8.2 8.2 8.2 8.3 8.2 8.2 8.2 8.2 8.3 8.3 8.3 8.3 8.3 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.4 8.3 8.3 8.3 8.3 8.3 8.4 8.3 7.2 7.2 7.2 7.3 7.3 7.3 7.3 7.3 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.5 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4 7.4
stattA' B' C* and 0 represent reP|icate test vessels containing five daphnids each at test
bSee last paragraph of D. made.
Test Methods for explanation as to why a single reading was
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STATIC. ACUTE, 48-HOUR EC5G O F H-19,519 TO D aphniam agna TABLE 3 C ontinued
Physical and Chemical Characteristics of H-19.519 Test Solutions
Nominal Concentrations
(mg/L)
Aa
Start .____________
Ba Ca Da
48 Hours __________ _ Aa Ba Ca
Oa
Temperature, c
H2O Control 20.1 b b b
15.6 20.1 b b b
31.3 20.1 b b b
62.5
20.1 b b b
125 20.1 b fa b
256
20.0
b
b
b
500 20.1 b b b
1000
20.1 b b b
20.5
20.5 20.4 20.3 20.5 20.3 20.3 20.4
20.5
20.5 20.4 20.3 20.4 20.3 20.3 20.4
20.5
20.5 20.4 20.3 20.4 20.3 20.3 20.3
20.5 20.5 20.4 20.3 20.4 20.3 20.3 20.4
aA, B, C, and D represent replicate test vessels containing five daphnids each at test . start.
bSee last paragraph of D. Test Methods for explanation as to why a single reading was made.
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Du Pont HLR 460-92
STATIC, ACUTE, 48-HOUR EC50 OF H-19,519 TO Daphnia magna
TABLE 4
Observed Immobility of Daphnia magna Exposed to H-19,519 for 48 Hours in a Static, Unaerated Test
Observed Immobility in Percent
Nominal Concentrations
(mg/L)
Aa
24 Hours Ba Ca
D*
48 Hours Aa Ba Ca Da
Water Control 0 0 0 0
15.6 0 0 0 0
3,1-3
0000
62.5
0000
125 0 0 0 0
250 0 0 0 0
5Q0 0 0 0 0
1000
0000
0000
0000 0000
0000 0000 0 O0 0 0000 0000
aA, B, C and D represent replicate test vessels containing five daphnids each at test start
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STATIC, ACUTE, 48-HOUR EC50 OF H-19,519 TO D aphnia magna
APPENDIX I
PROTOCOL AMENDMENTS AND SOP DEVIATIONS
PROTOCOL AMENDMENT NUMBER ONE
a. The definition of immobilization on page 8 of S O f j | ^ B H | L i i not be used In lieu thereof immobilization m il be defined as the inability to sw m ^at least two body lengths in any one direction) within 15 seconds after gentle prodding with a glass rod.
b. Daphnids w ill be distributed to test chambers using random numbers. No more than three daphnids will be placed into one chamber per random number.
c. A registered (not calibrated) American Society of Testing Materials mercury thermometer w ill be used to take temperatures of test solutions.
d. Analytical measurements of test solution concentrations will not be made.
Surviving daphnids at test end wilt be sacrificed according to S(
f. Solubility and stability studies of the test substance in dilution water w ill not be performed.
Q
S- !n s c P B B B B K p n page four the sentence "a test is not acceptable if more than l 0% 6Tjne_aaprmias die or show abnormal behavior in either control" is ambiguous. The sentence should read "a test is not acceoiable if more than 10% of the total daphnids in all four replicate controls die or show abnormal behavior"
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.
i. Temperatures will be tten 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.
SOP DEVIATIONS None.
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