Document 9JRJDN6bexQzbowpx5n1Q6m8D
AR226-2975
THAOE SSCHET
SlUilyJ]!l STATIC, ACUTE, 48-HOUR EC50 OF
H-19,519 TO Daphnia magna S^aSaJBBsmssaaBi
Orsanisation for Economic Co-Operation and Development (OECD) Guideline for Testing Chemicals: 202
Author Keith B. Pierson, Ph.D. Study Completed On
Ju(y1, 1992 Performing Laboratory E. I. du Port de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine Elkton Road, Post Office Box 50 Newark, Delaware 19714
Laboratory Project tD Haskell Laboratory Report No. 460-92
Page 1 of 16
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Do Font MM 460-92
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to EPA PIFRA and TSCA Good Laboratory Practice
Standards (40 CFR 160 and 40 CPR 792), OECD Principlesof Good Laboratory
iar- ' (0(81)30(^0^). Annex J2). ai3d MAPF Japan Good Laboratory PnacUce
Star.
(59 NohSan No. 3850). Solubility and stability of the test substance in
dilution dvater and under storage conditions were not determined. Concentrations of ihe
test substance in dilution water were not measured. Protocol amendments and/or SOP
deviations appear in Appendix 8. 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 Nemoure and Company
Sponsor: STUDY DIRECTOR:
Du Pont Chemicals E. I. du Port de Nerno. rs and Company
^wfL &. Pf^fsU
Keith B. Pierson, Ph.D. Research Toxicotogist
Environmental Sciences
WX 8.3. 7tw&
Mo/Day/Year
3 .Company Sanitized. Poss not contain TSCA CBI
Du Font HLR 460-92
TABLE OF CONTENTS
Page Specific Country Requirements--...........................................................................................2
Goc^l^graoryP{actfcSjatemeiit..................-..........................................................,.........3 Tabte of Contsnts..................................."..............--.......--.--..--...............--..............................4 Genera! hfcrniayon.........................................................................................^......................5
oUniIu3iyM&w*>* *" '*>** *B^
Quairty Assurance Documentation...........................................................................................?
i. Purpose...........................................................-......--......................................................8
11 ReOTrete...........................................................................-........-.-.........-.....................8
lit. Materials and Methods...................................................................................................8
A. Test Substance ..........................................................................................................S
B. Dilution Water............................,..........-............-.........-...---.................................8
C.
Animal
Culture
8
--.....--.......--.--......--....--........................----..--.-.........
D. Test Methods ....--...............--.....--.....------........--... ............................8
E. Chemical Analysis--..........,,.............................-...--.--.....
....................9
JV. Results and Discussion...............,........................................-.............-- .....................9
V. Conclusion............--.......--....................,,..--..--.....................--............................10
V(. Vtl.
VIH,
Literature Cited.........-..--......................................----.....----.------....-...10
Tables.--...........................--.......................................................................................11 1. Chemical Characteristics of Haskeli Laboratory Well Water--....................... 11
2. Water Chemistry of the Dilution Water at Test Start .....--.......-......................12 3. Physical and Chemical Characteristics of H-19,519 Test Sotutions.............13 4. Observed Immobility of Daphnia magna Exposed to H-19,519 for
48 hours in a Static, UnaeratedTest....--- .--..,,..........................--................15 AI pPpreontdoiccoelsA..m...e..n..d..m...e.-n..t.s..--an..d...S...O..P....D..e-.v..i.a.t..i.o..n..s............--...-.....--.......--.......--....................--...--.......................1166
4
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GENEBAl.
Do Pent HLR 460-92
H-19,519
Contaminants;
CA$ RggistixNumfasc
Saaosfln
.Study Initiated/Compteted: laiJfeLStudM
iaitiajg^CfimelfiteA
Du Pont Chemicals E. S. du Pont de Nemours and Company WHmington, Delaware 19898
6-8-92/7-1-92
6-10-92/6-12-92
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Du Font HLR 450-92
STATIC, ACUTE. 48-HOUR EC50 OF H-19,519 TO Daphnia magna
SUMMARY
H-19,519 was not toxic to Daphnia magna neonates (less than 24 hours old) in an unaerated, static, 48-hourtest. Nominal soncentratSons were 15.6, 31.3.62.5,125, 250,500 and 1000 mg/L. The 48-hour EC50 could not be calculated because no immobilizes occurred at the highest concentration tested.
Work By:
^L^A^S<^^ Robert W. Peterson, B.A.
Aquatic Associate
^ / ^ / ^ MoyDay^ear
JeUffr^ey^T.f.TTu^r^n-JeSr.M._o_/D_ ayC/Y/^ezAagr
Aquatic Associate
THIS flEPORT SSAPPROVED FOR ISSUE.
^jf.4.P7^eis^
Keith B. Pierson, Ph.D. Research Toxicoiogist and Study Director Environmental Sciences
/ Tfltt-V WSL Mo/Day/Year
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Du Font HLR 460-92
QUALITY ASSURANCE DOCUMENTATION (H-19519)
DNgS of Inspection:
Conduct-6/10/92 Records, Report(s) - 6/24,25/92
nndinqs reported to:
Study Director-6/29/92 Management - 7/1/92
Reported by:
^JLmJ^AJ&I^S^LbnjLT
Kimberty B. B^bner
Quality Assurance Auditor
^f//^ Date
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Du Font HLR 460-92
I. PURPOSE
The goal of this study was to assess the acute toxidty of H-19,519 to Daphnia magna neonates less than 24 hours old.
II. RECORDS
Alt data and the final report will be maintained either at Haskell Laboratory for Toxicology and industrial Medicine (Newark, Delaware) or be archived at the Du Font Records Management Center (Wilmington, Delaware).
HI. MATERIALS AND METHODS
A. Test Substance The test substance is describsd under General Information. Each 1100 mL solution was prepared by weighing the appropriate quantity of H-19,519, adding it to dilution
water (see below) and stirring for 60 minutes. Ei-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 of the test substance through volatilization, i ne stability and soiubiiity of H-19,519 in dilution water (see below) and water storage conditions were not determined.
B. Dilution Water Water (Table 1) originated from the Haskell Laboratory well and flowed through
aquaria containing fathead minnows, Pimsphales promelas. prior to use in this daphnid 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 autoclaved filtered dilutjojLwaler hejd.aLapproximately^Q^C. Daphnids were fed two algal species, Ankistrodesmus falcatus and Selenastrum capricomutum. at a rate df 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
gwpa^Sania^D^^^^"18^081
Du Font HLR 460-92
yc Screw-top Erienmeyer flasks (262 mL; 8-cm maximum diameter; 12.5-cm soiution depth;
and 13-cm total height) closed with a valve assembly (Miniert) was used. Each flask
contained approximately 262 mL of test solution and was filled to the top to minimize
head space and thus limit loss of test substance from volatility. Four replicates were
used for each test concentration; each repiicate contained five daphnids (1 daphnid/40
mL) for a total erf 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
(mean 2Q.3C) and were unaerated. Daphnids were not fed during the test.
A photoperiod of 16 hours Sight(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 me 16 hour tight interval. Observations were made daily.
Dissolved oxygen (YSlModel 54A, yellow Springs Instruments, YellowSprings, OH), pH (Coming Model 610A, Coming Scientific Instruments, Medfield, MA). and temperature (registered Am-rican 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, and. i ali replicates of the water centre! 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 would not fit through the neck of the Ertenmeyer flasks. Total alkalinity (Hach kit, Hach Chemical Company, Loveland, CO), EDTA hardness (Hach kit, Hach Chemical Company, Loveland, CO), and conductivity (YSE model 33, Yellow Springs Instruments, Yeliow 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 durino the 48-hour test.
E. Chemical Analysis Concentrations of H-19,519 in test solutions were not measured.
IV. RESULTS AND DISCUSSION
h 48-hour rar.geflnding 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 all 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 immobilitsss
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Du Font HLR 460-92
ooccccuurrrreedd a(Tt athbeleh4ig).heTsht eco4n8c-ehnoturartEioCn5t0esctoeudl.d not be calculated b"et^cauussce no iimmmmooboimlitiiieess
V. CONCLUSION
H-19,519 was not toxto to Daphnia magna neonates (!ess than 24 hours old) in an
unaerated, static, 48-hour test.
'
VI. LITERATURE CITED 1. TOersgtainnigsaCthioenmfiocralEsc: onom20ic2.Co4-OAppreilra1t9io8n4.and Development (OECD). Guideline for
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Du Pont HLR 460-92
STATIC. ACUTE, 4B-HOUREC50 OF H-19,519 TO Daphnia magna
TABLE 1
Chemical Characteristics of Haskeii Laboratory Well Water1
Parameter
Concentration
Parameter
Concentration
BOD, mg/L COD. mg/L QOC.mg/L TOC2. mg/L Total Kjeldah! N, mg/L Ammonia N, mg/L Turbidity. NTU
Phenolics, mg/L Color, Co/Pt
Solids
Total 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 iron3, mg/L
Fluoride3
<2 <10
2.8 <0.5 <0.2 <0.05
1.6 <0.050 <5
<4 <0.100 <0.060 <Q.01Q
<O.Q01
<0.002 24.5
6.2 <0.010 <Q,020 <0.010 <0.025
0.025
<0.1
Lead, mg/L Magnesium. mo/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
Sutflde. mg/L
Thallium, mg/L Zinc4, mg/L Volatile priority
poilutants
Acid extractabie priority pollutants Base/neutral
priority pollutants
Pestjcides/PCBs
Organophosphate pesticides5, ug/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 <0.020
ND
ND6
ND ND6
<0.50
i0at of sample collection 30 December 1991 unless indicated otherwise, analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey;
^ate of sample collection 15 January 1992, performed at Du Pont Engineering Test Center, Newark, Delaware; ^ate of analyses 13 January 1992, analyses performed at Du Pont
Chemicals, Jackson Laboratory, Deepwater, New Jersey; ^Below the Published Method Detection Limit (BMDL); SAnalyses performed at Hazieton Laboratories, inc., Madison, Wl;
^one detected.
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Du Font HLR 460-92
STATIC. ACUTE, 48-HOUR SQ59 OF N-19,519 TO Daphnia magus
TABLE 2 Water Chemistry of the Dilution Water at Test Start
Total Alkalinity' (mg/L as CaCOa)
85
EDTA Hardness (mg/L as CaCOg)
85
Conductivity
((imhostem)
180
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Du Font HLR 460-92
STATIC, ACUTE, 48-HOUR EC50 OF H-19.519 TO Daphnia magna TABLE 3
Physical and Chemica! Characteristics of H-19,519 Test Solutions
---Nominal-.
Concentrations (mg/L)
Start
--- ----_.-._--,,--.-_--. --
M
B9
C8
Da
48 Hours
-.-----------_.,,---.._..-_._
Aa
Ba
C3
Da
Dissatved Oxvpen. mg/L
HgO Control
8.1
b
15.6
8.1
b
31.3 82.5 125 250 500
1000
8.0
b
8.1
b
8.1
b
8.0
b
8.0
b
8.0
b
DH
HgO Control
7.2
b
15.6
7.2
b
31.3
7.3
h
62.5 125 250 500
1000
7.3
b
7.3
t>
7.3
b
7.3
b
7.3
&
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
b
^
b
b
b
b
b
b
b
b
b
b
b
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
"A, B, C, and D represent replicate test vessels containing five daphnids each at test
start.
mad^ee. last paragraph of D. Test Methods for explanation as to why a single reading was
S3 company Sanitized. Does not contajn TSCA CBl
Du Pont HLR 460-92
STATtG. ACUTE, 48-HOUR C50 OF H-19,519 TO Daphnia magna TABLE 3 Continued
Physical and Chemical Characteristics of H-19,519 Test Solutions
Start
Nominal
Concentrations
(mg/L)
A
B3
C3
DS
48 Hours
AS
B9
C8
Qa
Temperature. C
HzOControi
15.6
31.3 62.5 125 25Q
500 1000
20.1 20.1 20.1 20.1 20.1 20.0 20.1 20.1
b
b
b
b
b
b
b
b
b
bob b
b
b
bob b
b
b
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
"A, B, C, and D represent replicate test vessels containing five daphnids each at test .
sxart.
^See last paragraph of D. Test Methods for explanation as to why a single reading was
nade.
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Du Font HLR 460-92
STATIC. ACUTE, 48-WOUR EC50 OF H-19,519 TO Daphnia magna
TABLE 4
Observed Immobility of Daphnia magna Exposed to H-19,518 for 48 Hours in a Static, Unaerated Test
Observed Immobility in Percent
24 Hours
Nominal
Concentrations
(mg/L)
A3
Ba
C
D^
48 Hours
A"
B"
Ca
Da
Water Control
0
0
0
0
15.6
31.3 'v^
62.5 125
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
250
0
0
0
0
500
0
0
0
0
1000
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
0
^A. B, C and D represent replicate test vessels containing five daphnids each at test start
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Du Font HLR 460-92
c
STATIC, ACOTE, 48-HOUR EC50 OF H-19,519 TO Daphniamagna
APPENDIX I
PPOTOCOL AMENDMENTS AND SOP DEVIATIONS
PROTOCOL AMENDMENT NUMBER ONE
SOHp^JfB*m*iU ^ -
a. The definition of immobilization on page 8 of
not be used.
In iieu thereof immobilization will be defined as the inability toswnqat least two body
lengths in any one direction) within 15 seconds after gentle prodding with a glass rod.
b. Oaphnids will be distributed to test chambers using random numbers. No more than three daphnids witi 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.
8. Surviving daphnids at test end will be sacrificed according to SOI
f. Solubility and stability studies of the test substance in dilution water will not be performed.
_^^^^^^ g. En SOFBHH------^on page four the sentence "a test is not acceptable if more
than_1Q%.cBJlQft3^SfflBi(SBorshow abnormal behavior in either control" is ambiguous. The sentence should read "a test is not acceptable 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.
j. 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 wiii be assessed by a continuously-recording probe which wil! 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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