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Du Font HLR 448-91
Study Title
STATIC, ACUTE, 96-HOUR LC50
TO FATHEAD HINNOVS (PiBephOaPTeJs_"_p_ro_n_e_1
Data Requirement
Organisation for Economic Co-Operation and Development (OECD) Guideline for Tesiing Chemicals: 203
Author Keith B. Pierson, Ph.D.
Study Completed On 2 July 1991
Performing Laboratory
Haskell
E. I. du Font de Nemours and Company
Laboratory for Toxicology and Industrial Elkton Road, Post Office Box 50
Newark, Delaware 19714
Medicine
Laboratory Project ID Haskell Laboratory Report No. 448-91
Page 1 of 16
^p^s^.^o-----"'""15""1
Du Pont RLR 448-91
GOOD LABORATORY PRACTICE STATEMENT
This study was conducted according to United States Environmental Protection Agency Good Laboratory Practice Regulations for FIFRA (40 CPR 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.
Submit: ter:
Sponsor:
E. I. du Font de Nemours and Company
Du Pont Chemicals
Study Director:
\ij4L ^. //'gUffgo>-J
K_e_i_th__B.__Pi_e/rson, Ph.D.
Research Toxicologis';
Environmental Sciences
2</ Twe Wl
Day/Mo/Year
D
Du Font HLR 448-91
QUALITY ASSURANCE DOCUMENTATION
i^VmWWfp0 STUDY: \^--------l--------yS|jTATIC. ACUTE ,--96-HOUR LC50 OF
illt 19,011 "
FATHEAD MINNOWS
1^ (Pxmephales fffomelas)
AUDITS:
Items Audited Protocol, conduct
Records, final report
Audit Dates 6/14/91
6/27,28/91
SHORT-TERM AUDIT REPORT NUMBER: P-1517
DATE FINDINGS REPORTED TO MANAGEMENT AN'.) STUDY DIRECTOR: 7/1/91
/^i^^-^ Reported by: '^/q^e,^
/aames Mackay II g
Quality Assurance Auditor
~7/^/9/
Date
Company Sarunzud. UQ^S not eyioam i SCA CBI
3 -
Du Font HLR 448-91
TABLE OP COOTEWTS
Page
Good Laboratory Practice Statement
2
................. Quality Assurance Documentation
3
................... Table of Contents
4
.......................... I. Summary
5
............................. II. General Information
6
....................... III. Purpose
8
............................. IV. Records
8
............................. V. ............................................................................................................................................................ VI. ..................... VII. ........................... VIII. ..................................................................................... IX.
Materials and Methods
8
A. Test Materials
8
B. Dilution Hater
8
C. Animal Culture
8
D. Test Methods
8
E. Chemical Analysis
9
F. Statistical Analysis
9
Results and Discussion
9
Conclusion
9
Literature Cited
10
Tables
11
1. Chemical Characteristics of Haskell Laboratory Well Water
11 .
.
2. Lengths and Weights of Control Fathead Minnows (Pimephales
promelas) at Test Conclusion
12
.^ o&----U||HHBi 3. Wator Chemistry of Baskell Laboratory Dilution Water at Test
Start
13 ._ '_-_......... ._ _, '
4. Physical and Chemical Characteristics
--------------^HaskeNllumber 19,011) TesTsolutions
14 .
.
.
.
.
Behavior of Fathead Minnovaa(Pimephales promelas) Exposed to
------------im^--------d Haskell Number 19,011) for 96
Sours in a Static, Unaerated Test
15
Mortality of Fathead MinnoH& (Pimephales prcmelas) Exposed to
. . . . . . . . . . . . . . f------------------------mHa ikell Number 19,011) for 96
. . . . . . . . . . . . . . lours in a Static, Unaerated Test
16
Company Sanitized. Does not contain TSCA CBS
4
Du Font HLR 448-91
I. SUMMARY
lfP------------las moderately toxic in a 96-hour, unaerated, static test using juvenile faffiead ninnovs (Piaephales proaelas). Noainal test concentrations were 7.8, 13, 22, 36, 60, and 100 ag/L. The 96-hour LC50 based on noalnal concentrations vas 19 ag/L with 95Z fiducial Halts of 15 to 25
mg/L.
^Uz^ vork by: C. L. Hall Technician
Z/jL^/
^(fe 4. ft'^^ ^ ^fi
R. V. Peterson Technician
W^^ ?/ Day/Mo/Year
^ Te ^f
Day/Ho/Year
Study
Director:
'\e^i. 4. ri4t^
Keith B. Pierson, Ph.D. Research Toxicologist Environmental Sciences
Jiif TM. ?1 Day/Ho/'Year
THIS REPORT IS APPROVED FOR ISSUE.
^i^ /&^ 6.
Keith B. Pierson, Ph.D. Study Director
1 -ju.if llll
Day/Mo/Year
Company Sanitized. Doss nol contain TSCA CBI
II.
Material Tested:
Medical Research Number:
GlW.r
'FORMATION
JUIH
|------1
Haskell Number: Haskell Laboratory Test Code:
19,011
T^^U
CAS Registry Number:
Du Font HLR 448-91
Other Codes: Synonyms: Composition;
Purity:
Sponsor: Test Requested By:
Study Schedule: Notebooks:
Not supplied by sponsor
Du Font Chemicals
E. I. du Font de Nemours and Compan:'
Vilmlngton, Delaware 19898
Font Chemicals
E. I. du Font de Nereours and
1007 Market Street Vilmington, Delavara 19898
Company
Study Initiation Date: 12 June 1991
Experimental Start Date: 14 June 1991 Experimental Termination Date; 18 June
Study Completion Date: 2 July 1991
1991
Company Sanitized Does sw&cavSasn JSCA CB|
Dictrlbutlodi
^
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CompanySanitized. Does not contafri TSCA CEti
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Du Font HLR 448-91
III. PURPOSE
The purpose of this study was to assess the acute toxiciry o: to juvenile fathead minnows (Piaephales pronelas).
IV. RECORDS
All data and the final report will be maintained either at Baskell Laboratory for Toxicology and Industrial Medicine (Newark, Delaware) or be archived at the Du Font Records Management Center (Vilnington, Delaware).
V. MATERIALS AND METHODS
\. Test Materials
__ Source and composition of the test
Information. Appropriate quantities of)
cribed under General 'ere weighed and
dissolved in dilution vateiWsee below )**H--the test
iria. Stability and
solubility offHBB------Jin dilut-on water were not determined.
B. Dilution Water
Water (Table 1) originated from the Haskell Laboratory well.
C. Animal Culture
. Fathead minnows (Pimephales promelas) were reared in continuously flowing
'well water at Haskell Laboratory. Adul. tish resided in 38 L aquaria at
approximately 25C and were fed frozen brine shrimp (Artemia sp., Golden Gate
Brand, Kordon Division, Novaiek, Inc., Hayward, CA) generally twice dally, once
on Saturday and not on Sundays. Eggs were deposited on polyvlnyl chloride
tiles coated with sand. Tiles with eggs were aerated and incubated at
approximately 25C in a water bath. Larvae were reared in 22 L aquaria at
approximately 22C and were fed newly hatched Artemia so. (Aquariun Products,
Glen Burnie, MD) generally twice daily on weekdays and daily on weekends.
Juveniles were held in 272-L circular, fiberglass tanks (49-cm depth; 84-cm diameter) at approximately 20"C and were fed newly hatched Artenia sp. and/or
frozen brine shrimp generally twice daily on weekdays and once on Saturday and
Sundays. Sickness, injury, and abnormality were not observed. identified by labels on z^uaria and test vessels.
Animals were
D. Test Methods [1]
Six test concentrations and a water control were used. concentrations were 7.8, 13, 22, 36, 60, and 100 ag/L. The 2) ranged from 1.0 to 1.6 cm in total length (mean 1.4 cm),
standard length (mean 1.2), and 0.0061 to 0.023 g in weight
The
.lead minnows were 38 days old at test start.
Nominal
control fish (Table 0.09 to 1.4 cm in (mean 0.015 g).
Company Sanitized. Does not contain TSCA c
Du Font HLR 448-91
Glass aquaria (23 [1) x 16 (v] x 14 en [h]( 5-L) containing 4 L of test
solution (12-cm depth) were employed. Ten fish were added to each replicate using random nurbers (one replicate per concentration; total 10 fish per test
concentration). Loading vas 0.038 g/L. Test solutions ranged 21.9C (mean 21.7C) and were unaerated. Fish were not fed 48 nor during the test. A photoperiod of 16 hours light (409-452 hours darkness was employed. Observations were Bade daily.
from ^1.4 to hours prior to lux) versus 8
Dissolved oxygen (YSI Model 54A, Yellov Springs Instruments, Yellov Springs, OB) and pH (Corning Model 610A, Corning Scientific Instruoents,
Medfield, Mw) were measured in the water control and in all test concentrations
before fish were added at the beginning of the test and daily thereafter. Total alkalinity (Bach Chemical Company, Loveland, CO), EDTA hardness (Bach Chemical Company, Loveland, CO), and conductivity (YSI Model 33, Tel-low Springs Instruments, Yellov Springs, OH) of the dilution water were measured at the
beginning of the test. Temperatures in the water control were taken daily with
an American Society of Testing Materials registered mercury thermometer.
B. Chemical Analysis
Samples for chemical analysis were not taken at the request of the study
sponsor.
F. Statistical Analysis
Median lethal concentrations and their associated 95X fiducial intervals were calculated using probit analysis [2].
VI. RESULTS AND DISCUSSION
A 96-hr rangefinder was conducted with five fish in 2-L test solution in a 2-L glass beaker at each test concentration. Total mortality occurred at 1000 and 100, 10% at 10, and Of at 1.0 and 0.1 mg/L, respectively.
Nominal concentrations in the definitive test were 7.8, 13, 22, 36, 60, and 100 mg/L. All chemical and physical parameters (Tables 3 and 4) ware within expected ranges. Behavioral observations are summarized in Table 5. The 24-, 48-, 72-, 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.
VII. CONCLUSION
Jfl^BB^BBIIMras moderately toxic in a 9^-hour, unaerated, static test
us rag juvenile faflrcad minnows (Pimephales promelas).
Company Sanitized. Does not contain TSCA CM
mi. Limuffon cmo
Du root flU 448-91
1. 2. 333 p.
OKCD Guideline for Testing of Ch--iclst 203, 4 April 1984. Pinney, D. J. 1971. Probit AnalylB. 3rd Bd. Cubridfe Unlv. Pru.
Company Sanitized. Does not contain TSCA CBI 10
TABLE 1
Du Font HLR 448-91
Chemical Characteristics of Haskell Laboratory Veil Water1
Parameter
Concentration
Paraneter
Concentration
BOD2, mg/L
COD, ng/L
DOC3, mg/L
TOC<, mg/L
Kjeldahl N, mg/L
Ammonia N, mg/L
Turbidity, NTU Phenolics, mg/L Color, Co/Pt Solids
total suspended,
Aluminum, mg/L
Antimony, mg/L Arsenic, mg/L Beryllium, mg/L Boron5, mg/L Cadmium, mg/L
Calcium, mg/L Chloride6, mg/L Chromium, mg/L
Cobalt, mg/L Copper, mg/L Cyanide, mg/L Iron6, mg/L Pluoride6, mg/L
<2
13
1.5 <0.5 0.2 <0.05 <1.0 <0.050
<5
mg/L
<2
<0.100 <0.060 <0.010 <0.001 <0.020 <0.002 25.0
6.3 <0.010
<0:020
<0.010 <0.025
0.053 <0.1
Lead5, mg/L
Magnesium, ng/L
MBAS/LAS, ng/L Mercury, ng/L Nickel, mg/L
Mitrite6, mg/L Nitrate6, mg/L
Phosphate6, mg/L
Potassium, og/L
Selenium, icg/L
Silver, mg/L
Sodium, mg/L
Sulfate6, mg/L Sulfide, mg/L
Thallium, mg/L Zinc5, mp/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
lDate of analyses 17 October 1990 unless indicated otherwise; analyses
performed at Environmental Testing and Certification Corporation, Edison, New
Jersey; ^ate of analyses 13 December 1990, analyses analysis performed at
Environmental Testing and Certification Corporation, Edison, New Jersey; 'Mean value; ^ate of analyses 24 July 1990, performed at Du Font Engineering Test Center, Newark, Delaware; 'Below the Published Method Detection Limit (BMDL); 6Date of analyses 9 November 1990, analyses performed at Du Font Chemicals, Jackson Laboratory, Deepwater, New Jersey; ''Date of analyses 23 October 1990, analyses performed at Hazelton Laboratories, Inc., Madison, VI; 'None detected.
Company Sanded. Does not contain TSCA CW.
11
Du Font HLR 448-91
TABLE 2
Lengths and Weights of Control Fathead Minnows (Pimephales proaelas) at Test Conclusion
Number
Total
Length (cm)
Standard Length (cm)
Vet Weight (g)
1
1..5
2
1.,3
3
1,,6
4
1.,6
5
1. 6
6
1. 4
7
1.,1
8
1. 0
9
1. 2
10
1. 3
1.,3
0.019
1.,2
0.012
1,,4
0.019
1.,4
0.023
1.,4
0.022
1.,2
0.015
0,,9
0.011
0.,9
0.0061
1,,0
0.0087
1,,1
0.011
Company Sanitized. Does not contain TSCA CBI
12
Du Font HLR 448-91
TABLE 3
Water Chemistry of Haskell Laboratory Dilution Water at Test Start
Total Alkalinity (ag/L as CaC03)
72
EDTA Hardness (mg/L as CaCO,)
83
Conductivity
(umhos/cm)
170
Gompany S a n i t i z e d . "Dooees nnnot contain TSCACBf
13
Du Font HLR 448-91
Noainal
Concen
trations (g/L)
Water Control
7.8
13
22
36
60
100
^ 1FABLE
Physica 1 and ChenjLeal Char acteri[sties Hiiskell Nu aber 1L9,011 ) T8STSolutii3ns
Time; (hours)
pH
Dis solve(1 Oxygen (ng /L)
0 24 48 72 96
0 24
48
72
96
0
-- 1
TCBperature ("C
24
48
72
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
21.4 21.7
M"
N
N
N
N
H
N
N
N
N
N
N
21.9
N
21.8
N
N
N
N
N
N
N
N
N
N
N
O Beans all fish had previously died; "N eans reading was not >ade.
Du Font HLR 448-91
TABLE 5
Behavior
afjFathead Minnows
'Haskell Number
(Pimephales
19,011) for
promelas) 96 Hours
i
t o f Exposed
n a Static^
naerated
Test
Nominal
Concentrations (mg/L)
Number Affected/Total Number Alive
24 Hours
48 Hours
72 Hours
96 Hours
Water Control 7.8
13.
22. 36. 60. 100.
0/10
1/W
6/10"
3/7-
1/1"
D D
0/10 1/10"
3/1011
2/lOb
3/7-
i/r
D
D
0/10 0/9 1/9
1/6-
D* D
D
0/10 0/9 0/8
6/6*
D D D
Lying on bottom; ''at the surface; cat least one fish was at surface, at least one fish svam 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; 'at least one fish was lying on bottom, at least one fish was hyperactive, at least one fish svam erratically, and at least one fish shoved
partial loss of "quilibrium; *all fish had previously died so behavior could not be observed; and hyperactive and shoved partial loss of equilibrium.
Company Sanitized. ""Dooees nnot contain TSCACBI
15
Du Pont HLR 448-91
TABLE 6
Mortality o^JFathead Minnows (Pimephales promelas) Exposed to 'Haskell Number 19,011) for 96 Hours In a Static'" "TJnaerated Test
Nominal
Concentrat-iIons (Bg/L)
Cumulative Mortality in Percent
24 Hours
48 Hours
72 Hours
9b Hours
H,0 Control 7.8
13. 22. 36. 60. 100.-
0
0
0
0
0
0
30
30
90
90
100 -
100
100
100
0
0
10
10
10
20
40 ,- 40
100
100
100
100
100
1.00
Company Sanitized. Does not contain TSCA CBj
16