Document By1JN5pR5qbonzG1rqoB3dJvJ
M A Z 6 - O S S *!
TOXICITY TO AQUATIC PLANTS
TEST SUBSTANCE________________________________________________
Identity: A mixture containing perfluorooctanesulfonate, which may also be referred to as PFOS, FC-95, or as a component of FC-603EF. (1Octanesulfonic acid) (CAS # 2795-39-3).
Remarks: The 3M production lot number is not noted. The test sample is FC-603EF. Current information indicates it is a mixture of 1.30% PFOS, 10% diethylene glycol butyl ether, 54.15% water, 22.5% ethylene glycol, 2.6% propanesultone foamer, 5% Alkylether sulfate, 0.1% N-(3Chloroallyl)hexaminium chloride, 0.9% Xanthan gum, 0.9% starch, 1.0% triethanolamine, and 0.05% tolyl triazole.
The following summary applies a mixture with incompletely characterized concentrations of impurities. Data may not accurately reflect toxicity of the fluorochemical component of the test sample.
METHOD:________________________________________________________
Method: OECD Guideline 201 Type: Acute static GLP: No Year completed: 1996 Species: Selenastrum capricornutum Source: In-house culture. Originally from ATCC, Rockville, MD Element basis: Algal cell counts (cells/mL) and average specific growth rates. Exposure period: 96 hours Statistical Methods: IC50 values determined by the Trimmed SpearmanKarber Method and Toxstat software package, Version 3.5. Analytical monitoring: pH and temperature.
Test Conditions:
Algal Nutrient Medium: Per OECD method. This nutrient medium provided all mineral nutrients essential for algal growth and also served as the diluent for all algal operations. The pH of this synthetic algal medium was 9.06.
Stock and test solution preparation: A 1000 mg/L primary stock solution was prepared by diluting 150 mg of test substance in 150 mL of standard algal nutrient medium. Aliquots were then added directly to dilution water in test vessels to create test solutions.
Exposure vessels: 125 mL borosilicate-glass Erlenmeyer flasks containing 50 mL of test solution and capped with foam stoppers.
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Agitation: Shaken continuously at 100 rpm. Number of replicates: 3 for exposure concentrations, 6 for blank control. Initial cell loading: 1.0 x 104 cells/mL (1.2 x 104 actual) Number of concentrations: five plus a blank control Lighting: Continuous 400 ft-candles using cool-white fluorescent lamps. Water Chemistry:
pH range (0-96 hours): 7.48 - 9.06 Test temperature range (0-96 hours): 24.2 - 25.8 C
RESULTS______________________________________________ ___
Nominal concentrations: Blank control, 31.25, 62.5,125, 250, and 500 mg/L
Algal Growth Response EC5o values
Exposure (Contact)
Hours 24 48 72 96
Cell-Count mg/L (95% C.l.) 139 (1 17-165 ) 153 (1 24-189 ) 188 (1 7 3 -2 0 4 ) 141 ( 1 2 3 - 1 6 2 )
Average Specific Growth Rate
mg/L (95% C.l.) 171 (1 4 6 -1 9 9 ) 290 (254 - 330) 275 (252 - 300) 294 (265 - 326)
Element values are based on nominal concentrations.
Reversibility of Growth Inhibition: Aliquots of the 500 mg/L test solution were diluted with algal medium and cultured for 7 days. Based on growth observed in the recovery phase, the effect on algal growth was found to be algistatic.
Remarks: Testing was conducted on the mixture as described in the Test Substance Remarks field. The values reported apply to that mixture and not the fluorochemical proportion alone.
C m Q L U S lQ N S ____________________________________________________________
The test substance exhibits a 96-hour ECso cell count value of 141 (123 - 162 mg/L) and a 96-hour ErC50 growth rate value of 294 (265 - 326) mg/L.
The 96-hour No Observed Effect Concentration (NOEC) is 31.25 mg/L for cell count and 125 mg/L for growth rate. This test substance was determined to be algistatic.
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133
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DATA QUALITY Reliability: Klimisch ranking = 2. This study meets the criteria for quality testing. However, sample purity was not properly characterized and the study lacks analytical confirmation of the amount of fluorochemical proportion in the solution. R E F E R E N C E S ____________________________________________________ This study was conducted by AScI Corporation, AScl-Duluth Environmental Testing Division, Duluth, MN at the request of the 3M Company, Lab Request number P 2 5 8 1 ,1996.
OTHER______________________________________ _____________
Last changed: 6/26/00.
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b e s t c o p y a v a il a b l e
AScI Corporaiion/AScl-Duluth E m ironnicuut Tesimi* Division
AScI Siuv ID# 5010-068
STUDY TITLE
GROWTH INHIBITION OF FC-603EF. LIGHT WATHR(TM) ATC 3%, AR-AFFF
TO GREEN ALGA (Selenastrum capricomuium)
DATA STANDARD OECD GUIDELINE 201
STUDY COMPLETED November 22, 1996
TESTING FACILITY
AScI Corporation/AScI-Duluth Environmental Testing Division
4444 Airpark Boulevard Duluth, MN 55811
Tel. No. (218) 722-4040 Fax No. (218) 722-2592
'V va-?-
y K- M STUDY IDENTIFICATION NUMBERS
..................... ... ' AScI Study ID# 5010-068 w:
"
pfe;3M Company Lab Request# P2581
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1.0 GENERAL INFORMATION
ASci CorjxiraUoiv'AScI- Duluth Environmental Testing Division
AScI Study ID# 5010 068
Study Title
Data Standard Sponsor
Sponsor's Representative
Sponsor's Lab Request# Testing Facility
Principal Investigator Project Director Testing Facility Director Definitive Test Dates Data Validation and Report Review
Growth Inhibition of FC-603EF, Light Water (TM) ATC 3%, AR-AFFF to Green Alga (Selenasirum capricornumm).
OECD Guideline 201.
3M Environmental Technology and Services. Building 2-3E-09, 935 Bush Avenue. St. Paul, MN 55133; TeLNo. (612) 778-7452.
Susan Beach, 3M Environmental Technology and Services, Building 2-3E-09. 935 Bush Avenue, St. Paul, MN 55133; Tel No. (612) 778-7452.
P2581.
AScI Corporation/AScl-Duluth Environmental Testing Division, 4444 Airpark Boulevard. Duluth, MN 558i 1; Tel. No. (218) 722-4040.
Todd McGuire Joe Amato
Donald Mount
November 4-8, 1996.
Alan Mozol
Report Signature^
Retention of Raw Data and Final Report Location of Raw Data and Final Report
Two years from the date study is completed.
AScI Corporation/AScl-Duluth Environmental Testing Division, 4444 Airpark Boulevard. Duluth, MN 55811; Tel. No. (218) 722-4040. |
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AScI StuJv ID* 5010 008
2.0 OBJECTIVE(S)
To determine the 4-d median effect concentration (ECjJ. the 4-d no observable effect concentration (NOEC), and algicidal/algistatic effect of the test substance. FC-603EF. Light Water(TM) ATC 3%. AR-AFFF for green alga (Selenastrum capricomutum) under static test conditions.
This study was conducted according to OECD Guideline 201 (OECD 1993).
Test Substance Test Organisms
Algal Medium
3.0 TEST METHODS
Properties: Amber-colored liquid, specific gravity ca. 1.06 (water = 1). stable, and completely soluble in water.
Test concentrations: 0 (algal medium control). 31.23. 62.5. 25. 250. and 500 mg/L. Source: ATCC 22662
Inoculum preparation: Algal stock was subcultured in OECD medium for 4 days before test initiation.
Use in test: Stock culture was diluted and spiked into test solutions to obtain an initial cell density of 1.0 X 10* cells per ml (1.2 X 104actual concentration). Preparation: Deionized water was spiked with nutrient stocks (including Na2EDTA) to within OECD recommended concentrations. The most recent chemical analysis for deionized water is in Raw Data Package.
Characteristics: At test time, the medium had a pH of 9.06. Sterilized via autoclave prior to use in the test.
Exposure Chambers
Incubation
Observations
Endpoint Calculations
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Type: Sterilized 125-ml borosilicate-glass Erlenmeyer flasks sealed with foam stoppers.
Use in test: 50 ml of algal medium or test solution/chamber. Six control replicates and three replicates per test substance concentration were employed. During the test, kept stoppered, except when experimental observations were made. Solutions were not aerated during the exposures.
Duration: 4 days.
Daily photoperiod: Continuous at 400 ft-candles using cool-white fluorescent lamps. Test chambers held on shaker table set at 100 rpm.
Temperature: 25.0 2C.
Biological: Daily cell counts using direct microscopic enumeration via Improved Neubauer hemocytometer.
Water chemistry-: (1) At test initiation and termination- pH. conductivity, and temperature, (2) daily- temperature. All determinations were made according to APHA methods (1995).
1-, 2-, 3-, and 4-d EC5o's. and 4-d NOEC based on both cell counts and average specific growth rates (p). Algicidal/algistatic determination was | made post toxicity test termination.
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4.0 RESULTS
Test Substance Toxicity (Pertinent data are in Tables 1, 2, and 3)
Test Conditions
Cell Couni 1-d ECV1: 139 mg/L (117-165). P 1-d ECw: 171 mg/L (146-199).
Cell Counr 2-d E C 153 mg/L (124-189). fx 2-d ECm: 290 mg/L (254-330).
Cell Count 3-d CW: 188 mg/L (173-204). M3-d ECS0: 275 mg/L (252-300).
Cell Count 4-d EC.,: 141 mg/L (123-162). H 4-d ECW: 294 mg/L (265-326).
Cell Count 4-d NOEC: 31.25 mg/L. p 4-d NOEC: 125 mg L.
Effect of test substance was determined to be algistatic based on recovery of cells exposed to 500 mg/L concentration. Temperature ( C'): 24.2-25.8.
Lii>lit intensity ft-cdls: 400.
pH: 7.48-9.06.
Specific conductivity (pmhos/cnt): 137-152.
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5.0 TEST QA/QC
QA/QC Criteria
Algal counts in control must increase by at least a factor of 16 during the first 3-d of the exposure.
Test duration must be 4-d.
Data
During the first 3-d of the test control algal counts increased by a factor of approximately 39.
The test duration was 4-d.
6.0 REFERENCES
American Public Health Association (APHA). 1995. Standard Methods for the Examination of Water and Wastewater. APHA, Washington, D.C.
Organization for Economic Cooperation and Development (OECD). 1993. OECD Guidelines for Testing of Chemicals. OECD Publication Information Center. Washington, D.C.
Hamilton, M.A., R.C. Russo, and R.V. Thurston, 1977. Trimmed Spearman-Karber Method for Estimating Median Lethal Concentrations in Toxicity Bioassays. Environ. Sci. Technol. 11(7): 714-719; Correction 12 (4): 417 (1978).
West, Inc. 1996. Toxstat Software Package Version 3.5.
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TABLE 1. ECjo'S, 95% Confidence Intervals, and 4-d NOEC'S from Algal Growth Inhibition Test with FC-603EF Light Water(TM) ATC 3%, AR-AFFF
Endpoint Cell Count
Time t-d ECm 2-d ECm 3-d EC,, 4-d EC,, 4-d NOEC
Data*
(mg/L) 139 153 188 " 141 31.25
95 `i Confidence Interval (mg/L)
117-165 124-189 173-204 123-162 Not Applicable
Endpoint
Average Specific Growth Rate (/t)
Time l-d EC,, 2-d EC,, 3-d EC,, 4-d EC,, 4-d NOEC
Data*
(mg/L) 171 290 275 294 125
95 ci Confidence Interval (mg L)
146-199 254-330 252-300 265-326 Not Applicable
Values Based on Nominal Test Concentrations
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TABLE 2. Cells per ml and Percentage Difference from Mean Control Value for Selenasintm capricornutum Exposed to FC-603EF Light Water (TM), ATC 5%, AR-AFFF
Test Cone. (mg/L)
Control
31.25
62.5
125
250
`s v
500
Replicate
A B C* D E F Mean
A B C Mean
A B C Mean
A B C Mean
A B
c
Mean
A
B
C Mean
1d
8.5 (-) 8.0 (-)
- (-) 5.5 (-) 6.0 (-) 9.5 (-) 7.50 (-)
8.5 (13)7.0 (-7) 8.0 (7) 7.83 (4)
10 (33) 6.5 (-13) 5.5 (-27) 7.33 (-2)
3.5 (-53) 1.0 (-87) 4.0 (-47) 2.83 (-62)
3.0 (-60) 2.5 (-67) ; 2.5 (-67) 2.67 (-64)
v
2.0 (-73) 0 (-100) . 3.0 (-60) v. 1.67 (-78)
Cells/ml (10*)
2d 3 d
24 (-) 25 (-)
- (-1 28 (-) 37 (-, 27 (-) 27.7 (-)
49 (-) 48 (-)
- <-) 39 <-) 55 (-) 43 (-) 46.4 (-1
17 (-39) 19 (-33) 22 (-21) 19.2 (-31)
19 (-31) 17 (-40) 22 (-22) 19.0 (-31)
20 (-30) 18 (-37) 15 (-48) 17.2 (-38)
52(12) 46 (-2) 41 (-13) 46.0 1-1
45 (-4) 45 (-4) 44 (-5) 44.3 (-4)
43 (-8) 39 (-16) 3S (-18) 39.8 (-14)
8.5 (-69) 5.5 (-80) 10 (-64) 8.00 (-71).
2.5 (-91) 1.5 (-95) 3.0 (-89) 2.33 (-92)
12 (-75) 10 (-78) 9.5 (-80) . . 10.3 (-78)
2.0 (-96) 1.5 (-97) 1.5 (-97) 1.67 (-96)
4J
6t)(-| 62 (-) - (-) 52 (-) 91 (-) 79 (-)
68.5 (-)
65 (-5) 60 (-13) 56 ( IS, 60.2 (-12)
49 (-29) 52 <-24) 54 (21) 51.7 (-25)
49 (-29) 46 (-34) 46 (-34) 46.7 (-32)
14 (-80) 20 (-72) 14 (-80) 15.7 (-77)
;
2.0 (-97) 0.5 (-99) 4.0 (-94) 2.17 (-97)
Control Replicate C Values Omitted as an Outlier
f f\{vr -.
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~Jkr' '
k Parenthetic Values = % Difference from Control Mean Value
* 0-d Mean Algal Count > -1.2 X 10* ' 'jV # # ? ,;.1.-
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TABLE 3.
Average Specific Growth Rates (|t) and Percentage Difference from Mean Control
Value for Selenastrum capricomutum Exposed to FC-603EF Light Water(TM) ARAFFF
Tcit Cone. (m g/L) Control
31.25 6 2 .5 125 250 500
Replicate
A B O D E F Mean
A B C Mean
A B C Mean
A B C Mean
A B C Mean
A B C Mean
1d
1 .9 5 8 <-) 1 . 8 9 7 <-)
- () 1 5 2 2 <-) 1.609 (-) 2.069 (-) 1 833 (-)
1.958 (7)1.76 4 (-4) 1 897 <4; 1 8 7 6 |2>
2 .12 0 (16) 1.689 (-8) 1.522 (-17) 1.8 10 (-1)
1.0 70 (-42) 0.182 (-100) 1.204 (-34) 0.859 (-53)
0 .9 16 (-50) 0 .73 4 (-60) 0 .73 4 (-60) 0 .79 9 (-56)
0 .5 11 (-72) 0 (-100)
0 .9 16 (-50) 0.329 (-82)
u - In Cell per ml L - In Celh/ml L.
' 7 t.-io . :
A ve ra g e Specific G ro w th Rate t/ir
2d
1 .4 8 7 (-) 1.5 0 8 (-)
- (-> 1 566 (-) I 707 i i 1.5 4 7 1-, 1 5 70 (-)
1.325 (-161 1.3 68 t 13) 1 454 i-7i 1.385 (-12)
1 381 (-12) 1.3 11 1-17] 1 443 t Si 1.381 (-121
1.394 (-11) 1.3 40 (1 5 ) 1 246 (-2!) 1.330 (-15)
0 .9 7 9 1-38) 0 .76 1 ( 52) 1.0 6 0 (-32) 0 .9 49 (-40)
0 .3 6 7 (-77) 0 .1 1 2 (-93) 0.45 8 (-71) 0.3 32 (-79)
3J
1 233 (-) 1 2 3 0 <->
-I I 1 156 ( ) I 2H . , 1 189 i ) 1 2 1 8 ()
1 256 .3) 1 21 2 . 1 i 1 173 ; 4. i :i5 o.
I 204 , i , 1 204 i 1 j 1 201 . 1. 1 203 i i .
1 IS9 ( 2 ) 1 160 . 5j 1 152 i-5) t . 167 i-4)
0 753 * 3S) 0 70 7 (-42) 0 690 (-43) 0 .718 (-41)
0 .1 7 0 ( 86) 0 .0 74 (-94) 0 .0 74 (-94) 0 110 (-91)
4d
0 978 ( ) 0 984 ( )
-1) 0 940 ) 1 082 i i I 045 ( I 1.011 ( )
0 998 i 1) 0 l) 7 6 i 3 1 0 96! ) 5. 0 . 3
0 9 2 7 I-S) 0 942 1-7) 0 952 i 6, 0 941 ( 7 )
0 9 2 7 (-8) 0 909 i 10) 0 90) (10) 0 915 I 9)
0 61-1 t-3^i 0 697 (31) 0.605 (-40) 0.6 42 (-36)
.
0 128 ( 87) 0 219 (-10 0 ) 0 301 (-70) 0 148 (-85)
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