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Du Pont HLR 448-91
Study Title STATIC, ACUTE, 96-HOUR LC50 OP
TO FATHEAD MINNOWS (Pimepha
Data Requirement Organisation for Economic Co-Operation and Development (OECD)
Guideline for Testing Chemicals: 203
Author Keith B. Pierson, Ph.D.
Study Completed On 2 July 1991
Performing Laboratory E. I. du Pont de Nemours and Company Haskell Laboratory for Toxicology and Industrial Medicine
Elfcton Road, Post Office Box 50 Newark, Delaware 19714
Laboratory Project ID Haskell Laboratory Report No. 448-91
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Du Pont HLR 448-91
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to United States Environmental Protection Agency Good Laboratory Practice Regulations for FIFRA (40 CFR 160) and/or TSCA (40 CFR 792). Areas of non-compliance are documented in the study records. No deviations existed that significantly affected the validity of the study.
Submitter: E. I. du Pont de Nemours and Company
Sponsor: Du Pont Chemicals
Study Director:
. P,
Keith B. Pierson, Ph.D.
Research Toxicologist
Environmental Sciences
2V T m /99/
Day/Mo/Year
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Du P o n t HLR 4 4 8 - 9 1
QUALITY ASSURANCE DOCUMENTATION
STUDY: AUDITS:
STATICi AUTE .96-HOUR LC50 OF
0 [TO FATHEAD MINNOWS Pimephales t? 'm l a s )
Items Audited Protocol, conduct Records, final report
Audit D a t es 6/14/91
6/27,28/91
SHORT-TERM AUDIT REPORT NUMBER: P-1517 D A T E FINDINGS REPORTED TO M A NAGEMENT AN') STUDY DIRECTOR: 7/1/91
Reported by:
7~
ames Mackay II ^ lity Assurance Auditor
Date
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Du Pont HLR 448-91
TABLE OP CONTENTS
Page Good Laboratory Practice Statement ................................ 2
Quality Assurance Documentation .................................... 3
Table of C o n t e n t s .....................................................
I . S u m m a r y ........................................................ ...
II. General Information ............................................ 5
III. P u r p o s e ........................................................ ...
IV. R e c o r d s ..................................
o
V. Materials and M e t h o d s ....................
g
A. Test M a t e r i a l s ............................................ ...
B. Dilution W a t e r ...............
8
C. Animal Culture ............................................ g
D. Test M e t h o d s ................................................ g
E. Chemical Analysis ........................................ 9
- P. Statistical Analysis ....................................... 9
VI. Results and Discussion ........................................
9
VII. C o n c l u s i o n ..............
9
VIII. Literature Cited .............................................. 10
IX. Tables ........................................ 1. Chemical Characteristics of Haskell Laboratory Well Water
1111
2. Lengths and Weights of Control Fathead Minnows (Pimephales
promelas) at Test Conclusion .............................. 12
3. Water Chemistry of Haskell Laboratory Dilution Water at Test
Start ..............................
13
Chemical Characteristics 0 ____
________
Haskell Number 19,011) Test solutions . . . . . 14
fathead Minnova(Pimephales promelas) Exposed to
__ Haskell Number 19,011) for 96
sours in a Static, Unaerated T e s t .......................... 15
6.Mortality of Fathead Minnojm (Pimephales promelas) Exposed to
_ Ha,kell Number 19,011) for 96
Sours in a Static, Unaerated T e s t .......................... 16
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Du Pont HLR 448-91
I- SUMMARY
on^nominal concentrations vas 19 * / L vith'W
= r ."
fiSc^l X i
Work by: Technician
313m . ?/
Day/Mo/Year
4. P / * c ^ &
R- V. Peterson Technician
u/p 1
2t Tute 9t
Day/Mo/Year
Study Director: r^u>H. 4. ^
Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences
T**e ?/
Day/Mo/Year
THIS REPORT IS APPROVED FOR ISSUE.
_ A &^6 $.
Keith B. Pierson, Ph.D. Study Director
w i y ,q9/
D^/Mo/Year
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II. CliNF.r FORMATION
Material Tested:
Medical Research Number:
Haskell Number: Haskell Laboratory Test Code:
19,011
CAS Registry Number:
Du Pont HLR 448-91
Other Codes: Synonyms: Composition:
None
Purity: Sponsor: Test Requested By:
Study Schedule:
Notebooks:
Du Pont Chemicals
E. I. du Pont de Nemours and Company
Vilmington, Delaware 19898
'
ini Pont Chemicals E. I. du Pont de Nemours and Company 1007 Market Street Vilmington, Delaware 19898
Study Initiation Date: 12 June 1991 Experimental Start Date: 14 June 1991 Experimental Termination Date: 18 June 1991 Study Completion Date: 2 July 1991
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Du Pont HLR 448-91 III. FURPOSB
8The purpose of this study was to assess the acute toxicity to juvenile fathead minnows (Pinephales pronelas).
IV. RECORDS
tne uu pont Records Management Center (Wilmington, Delaware).
Test Materials
V. MATERIALS AND METHODS
solu B. Dilution Vater
b lniailut-lon watier werte not ydetermisnteadb.illty and
Water (Table 1) originated from the Haskell Laboratory well.
C. Animal Culture
-vellFvaterdat1H a ^ n (^ phaIeS ^ elas> were reared in continuously flowing
ssSS
J
? .on Saturday and no o I ' s I n H ^ ' l ' Hayward* CA> generally twice daily, once
tiles coated with
Su"f?ys' .E*8 were deposited on polyvinyl chloride
approximately 25C " "a
*1
Juveniles were held ^ 57? r
?aily weekdays and daily on weekends,
froaen^rine shrimp1
"^ a n d ^ r e ' f e S ^ y '
Sundays. Sickness i n i u i ^ ^ d ^ r
0" weekdays * * once on Saturday and
identified b y ^ b e i s ^ ^ u a S a
bSerVed`
D. Test Methods [1J
Six test concentrations and a water control were used Mn.<n.i
standard
i T t o ' l 's13' ?2 ' 36>i6?' " d 100 Bg/L' Tk* Cntro1 flsh <Table t0 ln total len& th (mean 1.4 cm), 0.09 to 1.4 cm in
The 1 lead mi ^mean 1,2)> and 0.0061 to 0.023 g in weight (mean 0.015 g).
The ..lead minnows were 38 days old at test start.
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' Du Pont HLR 448-91
nw i aSS,i?Uarl!
111 x 16 M x 14 CB (hli 5-L) containing 4 L of test
i6pth/) Wer*
Ten s h were added to each replicate
using random numbers (one replicate per concentration; total 10 fish per test
7 ? dln5 Was -038 g/L- Test solutlons
from 21?4 to
,,,,9 .C <mean 21-7 C) and were unaerated. Fish vere not fed 48 hours prior to
h* tCSt' f photoperiod of 16 hours light (409-452 lux) versus 8
hours darkness was employed. Observations vere made daily.
S D r i ^ Sii?d T T , i YSI,Mdel 54A> Yellov sPrins Instruments, Yellow J),and P* (Corning Model 610A, Corning Scientific Instruments,
neaiield, Hn) were measured in the water control and in all test concentrations
Tof2ieafiSitW^ e ?ideJ *1
beginnlng of the test and daily thereafter.
alkalinity (Hach Chemical Company, Loveland, CO), EDTA hardness (Hach
Chemicai Company, Loveland, CO), and conductivity (YSI Model 33, Yellow Springs
Instruments, Yellov Springs, OH) of the dilution water were measured at the *
fc th? t6St; Te"Peratures *" the water control were taken daily with an American Society of Testing Materials registered mercury thermometer.
B. Chemical Analysis
sponsorPleS ^ chemical
ere not taken at the request of the study
F. Statistical Analysis
Median lethal concentrations and their associated 95* fiducial intervals
were calculated using probit analysis [2J.
VI. RESULTS AND DISCUSSION
A 96-hr rangefinder was conducted with five fish in 2-L test solution in a anH eaS \ 5 M t concentratIn > Total mortality occurred at 1000 and 100, 20* at 10, and 0* at 1.0 and 0.1 mg/L, respectively.
100 "al concentrBtions in the definitive test were 7.8, 13, 22, 36, 60, an,
100 mg/L. All chemical and physical parameters (Tables 3 and 4) were within
expected ranges. Behavioral observations are summarized in Table 5. The 24-,
48 ' ,. ! and 96_hour LC50s based on nominal concentrations and observed
mortalities (Table 6) were 26 (21-31), 26 (21-31), 20 (16-26), and 19 (15-25)
mg/L, respectively.
.
using juvenile fa
VII. CONCLUSION
moderately toxic in a 94-hour, unaerated, static test minnows (Pimephales promelas).
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viii. u n a u f f o u a m
Du P o n t 8L8 448-91
1. OECD Guideline for Testing of Cheaicslst 203, 4 April 1984.
2. Finney, D. J. 1971. Probit Analysis. 3rd Ed. Caabridge Univ. Press. 333 p.
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TABLE 1
Du Pont Blit 448-91
Chemical Characteristics of Haskell Laboratory Well Water1
Parameter
Concentration
BOD2, mg/L
<2
COD, mg/L
13
DOC3, mg/L TOC4 , mg/L
1.5 <0.5
Kjeldahl N, mg/L
0.2
Ammonia N, mg/L
<0.05
Turbidity, NTU
<1.0
Phenolics, mg/L Color, Co/Pt
<0.050 <5
Solids
total suspended, mg/L <2
Aluminum, mg/L Antimony, mg/L
<0.100
<0.060
Arsenic, mg/L
<0.010
Beryllium, mg/L Boron5, mg/L
<0.001 <0.020
Cadmium, mg/L Calcium, mg/L
<0.002
25.0
Chloride6, mg/L
6.3
Chromium, mg/L Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron6, mg/L
<0.010 <0.020 <0.010
<0.025 0.053
Fluoride6 , mg/L
<0.1
Parameter
Concentration
Lead5 , mg/L
Magnesium, mg/L
MBAS/LAS, mg/L
Mercury, mg/L
Nickel, mg/L
Nitrite6, mg/L
Nitrate6, mg/L
Phosphate6 , mg/L
Potassium, mg/L
Selenium, mg/L
Silver, mg/L
Sodium, mg/L
Sulfate6 , mg/L
Sulfide, mg/L
Thallium, mg/L
Zinc5 , mg/L
.
Volatile priority
pollutants
Acid extractable
priority pollutants
Base/neutral
priority pollutants Pesticides/PCBs
Organophosphate .
pesticides7 , ug/L
<0.005 3.5
<0.05
<0.00020 <0.020 <0.1
1.6 <0.1
1.8
<0.005
<0.010 8.1
4.8 <0.05
<0.01 <0.020
ND*
ND*
ND* ND*
<0.50
^ D a t e of analyses 17 October 1990 unless indicated otherwise; analyses performed at Environmental Testing and Certification Corporation, Edison, New Jersey; *Date of analyses 13 December 1990, analyses analysis performed at
nyironmenta! Testing and Certification Corporation, Edison, New Jersey; 3Hean
rnf5 S"16 ?f j*nflyses 24 July 199* performed at Du Pont Engineering Test
er Newark, Delaware; 5Belov the Published Method Detection Limit (BMDL);
ae f a"alyses 9 November 1990, analyses performed at Du Pont Chemicals,
Jackson Laboratory, Deepwater, New Jersey; 'Date of analyses 23 October 1990, analyses performed at Hazelton Laboratories, Inc., Madison, WI; None detected.
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l Du Pont HLR 448-91
TABLE 2
Lengths and Weights of Control Fathead Minnows (Pimephales pronelas) at Test Conclusion
Number
21
3 4 5 6 7
8
109
Total Length (cm)
1.5 1.3 1.6 1.6
1.6
11.. 14
1.0 1.2
1.3
Standard Length (cm)
11..23
1.4 1.4
11..24
0.9 0.9
11.. 10
Vet Weight (g)
0.019
0.012
0.019 0.023
0.022
0.015
0.011
0.0061 0.0087
0.011
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TABLE 3 Vater Chemistry of Haskell Laboratory Dilution Water at Test Start
Total Alkalinity (mg/L as CaCO})
72
EDTA Hardness (mg/L as CaC03)
Conductivity (limhos/cm)
83 -
170
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Du Pont HLR 448-91
TABLE 4
Physical and Chemical Characteristics of Haskell Number 19,011) Ti
Time (hours)
Nominal Concentrations (g/L)
Hater Control
7.8
13
22
36
60
100
pH Dissolved Oxygen (mg/L) ______________________ ___________ ________
0 24 48 72 96
0 24 48 72 96
7.5 7.2 7.7 7.8 7.9 7.5 7.3 7.7 7.8 7.9 7.5 7.4 7.7 7.8 7.9 7.5 7.5 7.7 7.8 7.9 7.5 7.5 7.8 7.8 D* 7.5 7.5 D D D 7.5 7.5 D D D
8.9 9.1 8.2 8.5 8.5 8.9 9.2 8.2 8.5 8.4 8.8 9.2 7.8 8.5 8.5 8.8 9.4 8.0 8.5 8.5 8.8 9.4 8.1 8.5 D 8.8 9.4 D D D 8.8 9.4 D D D
Temperature (C)
0 24 48 72 96
21.4 21.7 Nb N NN NN KN NN NN
21.9 21.8 21.7
N NN N NN N NN N NN N NN N NN
*D means all fish had previously died; bN means reading was not made.
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Du Pont BLR 448-91
TABLB 5
IBehavior of Fathead Minnows (Pimephales pronelas) Exposed tc__ Haskell Number 19,011) for 96 Hours in a Static^Tinaerated Test
Nominal Concentrations
(mg/L)
Number Affected/Total Number Alive
24 Hours
48 Hours
72 Hours
96 Hours
Vater Control 7.8
13.
22.
36. 60.
100.
0/10 1/10 6/10
3/7"
1/1
D D
0/10 1/10
3/10
2/10b
3/7
1/1
D D
0/10
0/9 1/9=
l/6d
D D D
0/10
0/9
0/8
6/6*
D D D
Lying on bottom; bat the surface; cat least one fish was at surface, at least one fish swam erratically, at least one fish was lying on bottom, at least one fish had partial loss of equilibrium, and at least one fish was hyperactive; dat least one fish was lying on bottom, at least one fish was hyperactive, at least one fish swam erratically, and at least one fish showed partial loss of "quilibrium; all fish had previously died so behavior could not be observed; and ^hyperactive and shoved partial loss of equilibrium.
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Du Pont BLR 448-91
TABLE 6
Mortality n Pathead Minnows (Pimephales promelas) Exposed tol __________ -------- [Haskell Number 19,011) for 96 Hours in a StaticTHJnaerated Test
Nominal Concentrations
(mg/L)
H20 Control 7.8
13.
22.
36. 60.
100.-
Cumulative Mortality in Percent
24 Hours
0 0 0
30 90
100 100
48 Hours
0 0 0
30 90
100 100
72 Hours
0 10 10
40
100 100 100
9b Hours
0 10 20
.. 40
100 100 1.00
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