Document DvMaE7zaLJ9v0v3bMp15pbXEo
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ACTIVATED SLUDGE RESPIRATION INHIBITION TEST
TEST SUBSTANCE
Identity: Perfluorooctanesulfonate, Lithium salt; may also be referred to as PFOS Li salt, FC-94, or FC-94-X. (1-Octanesulfonic acid, 1,1,2,2,3,3,4,4,5,5,6,6,7,7,8,8,8-heptadecafluoro-, lithium salt, CAS #29457-72-5)
Remarks: The test sample is a mixture of the test substance in water (approximately 24.5% test substance and 75.5% water). No calculations were made to adjust for the actual concentration of the test substance in the test sample.
METHOD
Method: OECD 209 Test type: Static acute GLP: No Year Completed: 1994 Analytical monitoring: pH and DO concentrations. Statistical methods: Not performed Test organism source: Activated sludge mixed liquor collected from the Metropolitan W astewater Treatment Plant, St. Paul, Minnesota.
Test conditions Dilution water: Millipore Milli-Q water Synthetic Sewage: Per OECD method Reference and test solution preparation: A stock solution of the reference substance, 3,5-dichlorophenol, was prepared per the OECD method. The pH of the reference solution was measured to be 7.4. The test solutions were created by direct addition of the test sample: final concentration of 1000 mg/L. Initial pH of the two 1000 mg/L test solutions was 7.1 and 7.2. Test vessels: One liter glass fleakers. Number of concentrations: 1 (done in duplicate) plus 1 reference control and 2 Blank controls Temperature: 14.7-16.8C Total Suspended Solids and pH for sludge on day of testing: Total Suspended Solids = 4.9 g/L (mixed liquor). Final test solution mixed liquor suspended solids = 1.92 g/L. pH of the activated sludge mixed liquor gave an initial reading of 6.4 and a final reading of 6.7. Element Basis: Respiration inhibition as determined by oxygen consumption.
001554
Method Remarks: Dissolved oxygen in the test solutions was quite low during the 30-minute exposure portion of the test. The technician realized the test reactors were not being sufficiently aerated after the 30-minute readings and adjusted the air flow. Because the 3-hour data is the most commonly used data, it was decided not to re-test due to this occurrence.
Test temperatures were not controlled during the test. The respiration rates were normalized to 20C for the calculations.
RESULTS
Nominal concentrations: Two Blank controls, one reference substance (10 mg/L 3,5-DCP), two 1000 mg/L test material solutions.
Analytical Methodology: Analysis of DO concentrations in all test solutions were performed using a dissolved oxygen meter.
Control response: Satisfactory
Remarks: Values reported are for the test sample. No calculations were made to adjust for the concentration of the test substance in the test sample.
CONCLUSIONS
The test sample containing 24.5% Perfluorooctanesulfonate, Lithium salt exhibited a maximum inhibitory effect of 35% upon respiration at a nominal test substance concentration of 1000 mg/L at 3 hours exposure. The 3-hour EC50 (respiration inhibition) is >1000 mg/L.
Submitter: 3M Company, Environmental Laboratory, P.O. Box 33331, St. Paul, Minnesota, 55133
DATA QUALITY
____________
Reliability: Klimisch ranking 2. This study lacks information on the sample purity, and the test was conducted at temperatures below those recommended by the method. Oxygen concentrations also were too low to allow an adequate measurement of oxygen uptake rate following the 30minute exposure portion. These 30-minute results were considered invalid for the study. It was concluded that, since the 3-hour results show acceptable toxicity, the 30-minute results almost certainly would have shown the same.
REFERENCES
001555
This study was conducted by the 3M Company, Environmental Laboratory, Lab Request number M1018, 2/22/94. O T H E R ________________________________________________ Last changed: 5/3/00
001556
I
EVALUATION OF FC-94-X BY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209
Results
The inhibitory effect of FC-94-X, the lithium salt of perfluorooctanesulfonate, on activated sludge respiration was evaluated using OECD Test # 209. After calculated temperature corrections, measurements following a 3-hours exposure to FX-94-X at 1000 mg/L, showed activated sludge oxygen uptake rates were inhibited by 34%. The reference substance, 3,5-dichlorophenol, at 10 mg/L caused an inhibition of 56%.
Discussion
Though this study had deficits, the analyst considers that the results are sufficient to assess the potential for wastewater treatment system inhibition by FC-94-X.
One study deficit was that oxygen concentrations were too low to allow an adequate measurement of oxygen uptake rate following the 30-minute exposure portion of the OECD #209 study. These 30-minute results are thus invalid and should be ignored. The 30-minute results are not critical to the usefulness of the study, however, because inhibition is almost always greater following 3-hours of exposure. If the 3-hour results show acceptable toxicity, the 30-minute results almost certainly would have shown the same.
A second deficit is that temperature control was not used. Measured test temperatures were lower than that recommended by the method by as much as 5.5 C. However, all tests were conducted within 1.5 degrees of each other. Also, prior to comparison of the oxygen uptake rates, calculations were made to adjust the rates to those likely to have occurred at 20 C. The adjustment used was the rule of thumb that metabolic rates of mesophilic microbial populations typically double for each 10 C increase in temperature.
July 22, 1998
001557
JNTERNAL CRRESP0NDENCE/3M
TO: ERIC A. REINER FROM: SUSAN A. BEACH SUBJECT: Review of OECD 209 and BOD/COD Test Results for FC-94-X DATE: July 8,1998 I have reviewed the results from the test of FC-94-X on the inhibition of activated sludge respiration Inhibition (OECD 209) and have a few comments. 1. Although the dissolved oxygen in the test solutions was quite low during the 30-mlnute exposure portion of the
test, it was adequate after 3-hours exposure. The technician realized the test reactors were not being sufficiently aerated after the 30-minute readings, and adjusted the air flow. Because the 3-hour data is the most commonly used data, it was decided not to re-test due to this occurrence. 2. At the time this test was run, the-laboratory did not have a procedure in place for controlling the temperature during exposure. The temperature of the test reactors fluctuated with the temperature in the hood. Because this was tested in February in Minnesota, it is not surprising that the test temperature was so low. The respiration rates were normalized to 20C for the calculations. 3. The software used to produce the data reduction sheets did not have an easy way to depict exponents, so the equation for the temperature-adjusted respiration rates (Ra) is difficult to understand. It is as follows:
Ra = R adjusted to 20C = __ R
Where 0 = 1.072 x = T (observed) - 20C
4. Even though the 30-minute data is in question and the test was conducted at lower than normal temperatures, the data are useful for the interpretation of the toxicity of FC-94-X to activated sludge. An average of 34% inhibition was seen at 1000 mg/L. We generally do not consider inhibition less than or equal to 15% to be significant when using this procedure due to the jnherent variability of the method.
As for the BOD/COD testing, we have been unable to locate the data at this time.
Please see me if you have an questions.
Reaards.
Susan A. Beach
001558
ENVIRONMENTAL LABORATORY ACTIVATED SLUDGE MIXED UQUOR
LR-M iO tS
Date 3 1 3 3 j W
Analyst &MXJ P a g e _ L of j_
Type T e st: OECD ACTIVATED SLUDGE RESPIRATION INHIBITION, # 209
ACTIVATED SLUDGE MIXED LIQUOR
Date Obtained cl\<&\clLl
Date Used
Mixed Liquor Suspended Solids (MLSS) M i g/L
MLSS In final test solution 1^
g/L
Initial pH
(1st test soln. prepared)
Comments ____________________________ _______
Adjusted to '
g/L
Final pH
(last soin, prepared)
TEST SUBSTANCE
Sample Description frCj~c\Li --X
' _____________________________________
Initial pH of highest cone, tested (lO C C f^ J ^ -) *7. <2
Adjusted to pH
Comm ents _____________________ ___________________________________________ ^______
REFERENCE SUBSTANCE
Sample Description * 5 j
H ^ ) \ C V A L n ( T ^ f N A p L ----------------------------------
Initial pH of stock solution ( ^>Q O mg/L) = "71*-]
Adjusted to pH
Comments _______________ _________________________________________________
EXPOSURE CONPTTIONS
Test Duration
3 \Aoa>E-S______________________________________________
Test Temperature Range:
14.C
Comments _________________________________________________________________
.................
001559
ENVIRONMENTAL LABORATORY
lr JA ttA S
OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 TEST SOLUTIONS PREPARATION
Test Substance:
-- R U --y _______________________
Sample Description: `"fogCt VPu^n
Analyst:
*
Date: c p j ^ J
________
Test Cone. mg added
ma/L
per 500 mL
W C kJW L -
mL M o i1) water per 500 mL
aw
mL syn. sewage feed oer 500 mL
Jb
mL microbial inoculum oer 500 mL
Total Volume mL
______2 C \ ________ ______ S M
/O O P ____S D Q ____
a y j ______ ________ L L _______ ______ 2 m _______ _______s r o c s
IC O O
5 O
_____ m ______
lb
<500
J * Ti&t. htfay fi
C oaJ C i 7o >t Soin u /u
iiOiTtriL.
_____ ______________________________ _____________ i : ___________________________________
100
/e r r )
____________ ~7i \
(1) Miilipore Milli-QR high purity water
001560
ENVIRONMENTAL LABO RATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209
RAW DATA
^
T e st S u b sta n ce : P C " 0 ! H - X ______ '
____________________________________
Source of Activated Sludge Mixed Liquor: ll/Jv.- L()aS>TR
yci S p fltuL
Date Received:
Date Used:
A?V
Analyst: ' T u O
30-MINUTE EXPOSURE
Test Substance
Concentration (/^c ) 0.5
1.0
DISSOLVED OXYGEN CONCENTRATIONS, mg/L
TIME-MINUT>C*O 1.5 2.0 2.5 3.0 3.5 4.0 4.5
5.0
Temp
VcaMc CC
EK CONTROL 1
WOO /CO EK CONTROLH
7 0 fc/i
(,.Q
s.H 5 .0 4 ,7 - 4 .5 7 .0
(S
!> i , r
I <? 1 , 0 1 0 . O -S --
n.C ? o s ' -
! 1-
----
1
1
----
----
-- H U IT
-- Ik ,5
6 .0 2 1 .
(* S 12:2- S',7 .C> S -7 S'.! IS".
3-HOUR EXPOSURE
DISSOLVED OXYGEN CONCENTRATIONS, mgrt-
Test Substance Concentration
TIME MINUTE3
Temp
) 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0 5.5
c
EKCONTROL1
/r-c /cct>
EKCONTROLII
7 ,e
1
IC.G
'1
f.3
'll y 7 / L,1 k , r 6 3 L,o S .7 s W s'. I
O.'J G.O 5 e! 5 7 5 ^ _ 5 i3 5 . \ 1
5s \ 7,7 7.7- 7,r 7.3 7.1 3fl
1
1
<7,7 7.7" 7,r
5 7 3 5 " 3 5
/< /,? /k ,0 i5 i
77 i L _ L? L?,S"" G?<5 [a>0 5.7 .7 6 . 3
17,5
pH of activated sludge mixed liquor: Initial
pH of highest concentration tested:
Initial
Test tem perature: Id. 7 - K ^ .g 0^ .
______ .5
Final (>,*7 Final t,A
001561
ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 CALCULATION OF RESULTS
Test Substance: FC-94-x
Analyst: RJW
Ln M1018
Date: 2/22/9.4
^ 4o < r4 .
sD
H
30-Mlnute Ex oosure m g/L 0.5 1.0
BK1 7.0 6.9
Dissolved Oxygen Concentrations, mg/
1.5 2.0 2.5 3 .0 3 .5 4 .0 4.5 ,5 .0 5.5 Temp, ` C
WBSBBBM
l l l l t l WSBBBBBt
6.4 6.0 5.7 5.4 5.0 4.7 4.3 4.0 3.6
16.6
R (D 0.7079
Ra (2) % Inh. (3) i! p ; |i |
0.8966 6
1 0 0 0 .0 2.3 1.7 1.5 1.2 1.0 0.8 0.5
16.8 0.4743 0.5925
31
1 0 0 0 .0 1.2 0.9 0.6 0.5
16.8 0.4000 0.4997
41
BK2 8.0 7.7 % cliff in controls (4):
7.4 10.1
7.1
6.8 6.5 6.2 5.9 5.6 EC50 (95% Cl): > 1000 mo/L
5.4
5.1
15.2
0.5806 0.8106 O RaCm (5): 0 .8 5 3 6
m g/L 0.5 1.0 1.5 2 .0 2.5 3 .0 3.5 4 .0 4 .5 5 .0 5.5 Temp, *C R (1> Ra (2) % Inh. (3)
1II1M I
P ^ il
iP ljP llll
BK1 7.9 7.6 7.4 7.1 6.8 6.5 6.3 6.0 5 .7 5.4 5.1
14.7
0.5588 0.8078 6
10 0 0 .0 6.6 6.3 6.0 5.9 5 .7 5.5 5 .3 5.1 4.9 4 .7 4.5
16.0 0.3915 0.5170
35
1 0 0 0 .0 5.3 5.1 4.9 4.7 4.5 4 .3 4.1 3.9 3.7 3.5 3.3
15.9 0.4000 0.5319
33
BK 2 7.4 7.1 6.9 6.5 6.2 6 .0 5 .7 5.4 5 .2 4 .9 4 .6
% cliff in controls (4): 2 .3
EC 50 (95% Cl): > 1000 mo/L
14.9
0.5539 0.7897
_____
RaCm (5): 0 .7 9 8 7
(1) FU Oxygen uptake rate (mg 02/L-m ln) (2) Ra R adjusted to 20' C= [R/e.Ex] (3) % Inhibition [(RaCm)-RaS/(RaCm)]100 (4) % difference In controls: [IRaBk1-RaBk2l/RaCm]100 (Two controls must be within 15% of each other.) (5) Mean adjusted R, controls [RaBk1+RaBk2]/2
0 1.072 RaCm mean adjusted R, controls Bks blank control. S * sample Tobs= sample temp x Tobs-20*C
ENVIRNOMENTAL LABORATORY
lr lotsa- mim
^ \0 F 3
OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 TEST SOLUTIONS PREPARATION
Test Substance: S '- \CM
^ V \l/\C L 'H o W l /)C __ u b S T T W c < r
Sample Description:
L ric x A L e t aJO 0% oL\D~DWl_________________________ _
Analyst: ``T R S lO _________
Date:
________
Stock Solution Preparation: T )'< ,y > W d
A O O E ft -- AT)TM?P> T V L o re P
ac>\ \ m l -
r r v j____________________________ _
^ m < rvS r 3>fS~- T bC P \/Q
\Q m i \ ,CTnJ A JaO H
3 0 fW T m n uJqhM .
fO V A ^S oi/ ^ TM nyLO vsE- i)N )h g & &. v tz g i^ fr o n n L
P P T7
-Vt> \.Q L i TF f^___lOSTH M q U > \T g ft_______________________________
y v \ * 7>H
Test Cone. mL added mL MOO ) water mL syn. sewage
ma/L
Der 500 mL
per 500 mL
feed oer 500 mL
'BJCCa t v 'l --
____ ___________ ______ Z k ________
mL microbial inoculum Der 500 mL
200
Total Volume mL
SOO
/
ID ____ 2 2 ! i______
/k?
<200 '
TOO
(1) Millipore Milli-Q^ high purity water
001563
ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209
RAW DATA
3 0 3
Test S ubstance : 3 ^ 5 " ^ \o - P r .p W y v .I ,, `'ReSe/PnCG--
________
Source o f Activated Sludge Mixed Licuor: AULtnIjL W f l / W r I /r a 'W v iT ' P I <WT~ ST,
s//aJ
Date Received:k l i i l l
Date Usad: ^ I
'^ Y S L
30-MINUTE EXPOSURE
Test Substance
Concentration falL. ) 0 .5
1.0
BK CONTROL 1
ib
1,0 T. b 1KM
1
DISSOLVED OXYGEN CONCENTRATIONS, mg/L
TIME MINUT c a 1.5 2.0 2.5 3.0 3.5 4.0 4.5 5.0
Temp 5.5 C
U ,0 r . T 5.H .0 - 4 , 7
2,1 1 7 , 7 7 ^
4 .3 M.0 3 .0 Ib.b
7 5 . 7.0
jyy-
EK CONTROL II
1 1 1
? , 0 7 , 7 71A 7 . | 6 . * ( o C?. 3
1 1
", G? . i L 5 ,1 IS.SL
3-HOUR EXPOSURE
DISSOLVED OXYGEN CONCENTRATIONS, mg/L
Test Substance
TIME - MINUTES
Temp
Concentration { Afy/c ) 0 .5 1 .0 1 .5 2.0 2 .5 I 3.0 3 .5 4 .0 4 .5 5.0 5.5
C
EK CONTROL 1 IO
7,1 7C? 7.H 7,!
o . S (jfO 5 . 7 - S W < T / /V .7 -
8.3. S ,\ 3 .0 7,1 7,? r7.T i . r 7 W 7,3 7 . 3 7.0 i4f.r
1
1
EK CONTROL II
7*7 7 , )
pH of activated sludge mixed liquor: pH of highest concentration tested: Test tem perature: N . S'- k 0
i- i . a
Initial (/H Initial
& /
*4 5 .3
Final. L,
Final Ls><f
001564
001565
ENVIRONMENTAL LABORATORY OECD ACTIVATED SLUDGE RESPIRATION INHIBITION TEST #209 CALCULATION OF RESULTS
Test Substance: 3.5-Dichlorophenol Reference Substance
Analyst: RJW
30-Mlnute Ex sosure
Dissolved Oxygen Concentrations, mg/l
mg/L 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 , 5.0 5.5
fc l
iiili
' 's i iMlli
BK1 7.0 6.9 6.4 6 .0 5 .7 5.4 5 .0 4 .7 4 .3 4 .0 3 .6
10.0 8.6 8.4 8.3 8.1 7.9 7.7 7.5 7.3 7.2 7.0 6.8
LH Midi A MlUfl
Date: 2 /2 2 /9 4
"P j 3 oP 3
Temp,*C R, O) R a (2) % Inh. (3)
16.6
0.7079 0.8966
14.7
0.3636 0.5257
38
BK2 8.0 7.7 7.4 7.1 6.8 6.5 6.2 5.9 5.6 5.4 5.1
% dift in controls (4): 10.1
EC50 (95% Cl): > 10 mo/L
15.2
o0 .5 8 0 6 0 .8 1 0 6
RaCm (5): 0 .8 5 3 6
m ^/L 0.5 1.0 1.5 2.0 2.5 3 .0 3.5 4 .0 4 .5 5 .0 5.5 Temp, *C
liyi||l
iM H I
m ik
W M i B i l
R (1) Ra (2) % Inh. (3) II I m ;
BK1 7.9 7 .6 7.4 7.1 6.8 6.5 6.3 6.0 5 .7 5.4 5.1
14.7
0.5588 0.8078 6
10.0 8.2 8.1 8.0 7.9 7 .8 7.7 7.5 7.4 7 .3 7.2 7.0
14.5
0.2412 0.3536
56
BK 2 7.4 7.1 6.9 6.5 6.2 6.0 5.7 5.4 5.2 4 .9 4 .6
% diff in controls (4): 2 .3
EC 50 (95% Cl): < 10 mo/L
14.9
O___0.5539 0.7897
RaCm (5): 0 .7 9 8 7
(1) R Oxygen uptake rate (mg 02/L-m ln) (2) Ra R adjusted to 2 0 * 0 [R/e.Ex] (3) % Inhibition [(RaCm)-RaS/(RaCm)]100 (4) % dRterence in controls: [IRaBk1-RaBk2l/RaCm]100 (Two controls must be within 15% o l each other.) (5) Mean adjusted R. controls [RaBk1+RaBk2]/2
e - 1.072 RaCm mean adjusted R, controls Bk= blank control, S sample Tobs sample temp x - Tobs-20*C
