Document Yjmapg15BeYGjB0ppz3MQe4mO
BACK TO MAIN
Study Title
Characterization Study EtFOSE-OH, Test Control Reference #SE-035 PHASE: SOLUBILITYOF ETFOSE-OHINWATER, METHANOLA,ND ACETONE
Data Requirement
40 CFR 160.105(b)
Author
Mark E. Ellefson
Phase Completion Date
Date of signing
Performing Laboratory
3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue
St. Paul, MN 55106
Project Identification
3M Environmental Laboratory Study # FACT-TCR-004 (LIMS #E00-1715) Centre Analytical Laboratories Study # 023-023
Total Number of Pages
258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
~~~
~
~
~~~
GLP COMPLIANCSETATEMENT
Study Title: Characterization Study EtFOSE-OH, Test Control Reference #SE-035, Phase: Solubility of EtFOSE-OH in Water, Acetone, and Methanol.
Study Identification Number: FACT-TCR004, Centre Analytical Laboratories Study # 023-023
This phase of the study was conducted in compliance with Environmental Protection Agency (EPA) Good Laboratory Practice (GLP) Standards 40 CFR 160 with the exceptions in the bulleted list below.
Exceptions to GLP compliance:
0 The electronic data systems in use have not been validated and there is not an electronic audit trail of corrections currently available (40 CFR 160.130 (e)). Authenticated hardcopies of chromatograms and associated documents will be considered as the original raw data.
0 The substance used for solubility testing was depleted and another lot was used for solubility in water. The purity of this second lot was verified only after its use in the study.
0 The labels on containers of test substances initially did not show expiration date or storage conditions.
/
William K. Reagen, Testing Facility Management
Dde
0J-/3o/U/
Date
LlMS #E00-1715,Page 2 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
QUALITY ASSURANCSETATEMENT
Study Title: Characterization Study ETFOSE-OH, Test Control Reference #SE-035 PHASE: Solubility of EtFOSE-OH in Water, Methanol, and Acetone
Study Identification Number: FACT-TCR004, Centre Analytical Laboratories Study W 023-023
This phase of the study has been inspected by the 3M Environmental Laboratory Quality Assurance Unit (QAU) as indicated in the following table. The findings were reported to the study director and laboratory management.
I 09/08/00
02/0527/01
I-05117-18/01
I Solubility Screen Test
Data Draft Phase Report
02128/01 05/23/01
02128/O1 05/23/01
<& d w /
Date
QAb Reprdsentative
LlMS #E00-1715, Page 3 of 258
BACK TO MAIN
3 M Environmental Laboratory Report No. FACT-TCR-004
TABLEOF CONTENTS
GLP Compliance Statement............................................................................................................... 2 Quality Assurance Statement............................................................................................................. 3 Study Information ............................................................................................................................... 6 Summary ............................................................................................................................................. 7 Purpose .............................................................................................................................................. 7 Test Substance .................................................................................................................................. 8 Reference Substance......................................................................................................................... 8 Test System ....................................................................................................................................... 9 Method Summaries ............................................................................................................................ 9 Preparatory Methods.......................................................................................................................... 10 Sample Collection and Analysis......................................................................................................... 12 Analytical Method ............................................................................................................................... 16 Analytical Results............................................................................................................................... 17 Data Summary and Discussion.......................................................................................................... 18 Statistical Methods ............................................................................................................................. 22 Statement of Conclusion.................................................................................................................... 22 References......................................................................................................................................... 22 List of Attachments............................................................................................................................. 22
.
LlMS #E00.1715. Page 4 of 258
BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
LISTOF TABLES
Table l a. Summary Table of Solubility of ETFOSE-OH in Water...................................................... 7 Table Ib. Summary Table of Solubility of EtFOSE-OH in Methanol and Acetone............................ 7 Table 2. Characterization of the Test Substance and Analytical Reference Substance................... 8 Table 3. Characterization of the Analytical Reference Substance .................................................... 8 Table 4a. Description of test systems used in Solubility Screen Test .............................................. 9 Table 4b. Description of test system used in Solubility Test. Column Elution Method ..................... 9 Table 5. Summary of Screen Test method Sample Preparation...................................................... 10 Table 6. Column Elution Method Preparation for Study (FACT-TCR004). ETFOSE-OH TCR-
00017-98 at 22 rpm and 11rpm.......................................................................................... 12 Table 7a. Sample Collection. Preparation. and Analysis for Study FACT-TCR004 EtFOSE-OH 22
rpm flow rate........................................................................................................................ 13 Table 7b. Sample Collection. Preparation. and Analysis for Study FACT-TCROO4 EtFOSE-OH 11
rpm flow rate........................................................................................................................ 15
Table 8. Calibration curve ranges used for the quantification of EtFOSE-OH 11 and 22 rpm data..17
Table 9a. Individual EtFOSE-OH sample concentrations for 22 rpm study...................................... 19 Table 9b. EtFOSE-OH sample concentrations by timepoint for 11 rpm study.................................. 20 Table 10a. Summary Table of Solubility of EtFOSE-OH in Water..................................................... 21 Table 10b. Summary Table of Solubility of EtFOSE-OH SE-035 in Methanol and Acetone............. 21
.
LlMS #E00.1715. Page 5 of 258
BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
STUDYINFORMATION
Study Director
John Flaherty Centre Analytical Laboratory 3048 Research Drive State College, PA 16801
Analytical Chemistry Laboratories Solubility Testing
Certificate of Analysis
3M Environmental Technology and Safety Services (ET&SS) 3M Environmental Laboratory Bldg. 2-3E-09 935 Bush Avenue St. Paul, MN 55106
Centre Analytical Laboratory 3048 Research Drive State College, PA 16801 John Flaherty, Study Director
Dr. William K. Reagen, Labomtory Manager Mark E. Ellefson, Principal Investigator Kristin L. Terrell, Analytical Chemist Mark L. Anderson, Analytical Chemist Cindy M. Carlson, Analytical Chemist
Sponsor
3M Environmental Laboratory 935 Bush Avenue Building 2-3E-09 St. Paul, MN 55106
Dr. William K. Reagen
Experimental Dates
Experimental Start Date: 11 September 2000 Experimental Termination Date: 09 January 2001
-
Archives
Reserve samples of the reference standards will be maintained by the 3M Environmental Laboratory. All study data for this phase and a copy of the phase report are archived by the 3M Environmental Laboratory.
LlMS #E00-1715, Page 6 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
SUMMARY
Solubility of EtFOSE-OH in water is a significant parameter because the spatial and temporal movement (mobility) of a substance is largely determined by its solubility in water. Water soluble substances readily gain access to humans and other living organisms; furthermore the knowledge of the solubility in water is a prerequisitefor testing biologicaldegradation, bioaccumulation, and other tests. Solubility of EtFOSE-OH in methanol and acetone is a significant parameter for
general laboratory knowledge in the preparation and analysis of EtFOSE-OH. The approximate
solubility of EtFOSE-OH in water was determined by performing a solubility screen test (ETS-8170.0), followed by a quantitative solubility determination via the Column Elution Method (ETS-8171.O) and HPLC/MS analysis (ETS-8-155.0). The approximate solubility of EtFOSE-OH in acetone and methanol was determined by performing a solubility screen test.
Results are presented in table l a and 1b:
Table la. Summary Table of Solubility of ETFOSE-OH inWater.
22 rprn study
11 rprn study
I Finalvalue I
159
10,6%
5 timepoints
I42
6,4%
5 timepoints
151
I
11 % RPD
I 2consecutlveruns I
The solubility of ETFOSE-OH TCR-00017-98 in water was determinedto be 151 n g h L (corrected for purity) at 19.5-21 "C.
Table 1b. Summaty Table of Solubility of EtFOSE-OH in Methanol and Acetone.
I
Methanol
I
5 10%
I
Acetone
> 10%
The solubility of EtFOSE-OH was determined to be greater than 10% (masslvolume) in methanol and acetone at 22.5"C.
PURPOSE
The purpose of this phase of the study was to determine the solubility of EtFOSE-OH in water, methanol, and acetone as part of the characterizationof this test, control, and reference substance.
LlMS #E00-1715, Page 7 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
TESTSUBSTANCE
Table 2. Characterization of the Test Substance and Analytical Reference Substance
3M Laboratory Identification Number Source
Expiration Date Storage Conditions Physical Description
Purity
SE-031
Wildlife
International
11-26-2001
Frozen
Yellow solid (sandy) 97.7%
SE-035
Puritiedfrom SE-031 at Pace, Tier II
08-17-2001
Frozen
White Solid
99.9%
TCR-00017-98
Purifiedfrom SE-031 at Tier 2 Facility
Not Available
Frozen
White powder
99.9% (unofficially)
EtFOSE-OHTCR-0017-98 was used for the column elution method instead of SE-035 because there was limited supply of SE-035 required to complete the entire study. Since EtFOSE-OH TCR-0017-98 was purifiedfrom the same source (SE-031) and method as SE-035, it was used as a suitable replacementfor the quantitativedeterminationof the EtFOSE-OH solubility. The purity of the reference substance, SE-035, has been determined to be 99.9%. All reported values have been corrected for purity.
REFERENCSEUBSTANCE
Table 3. Characterization of the Analytical Reference Substance
sour&
Expiration Date
Storage Conditions
3M Laboratory
- Identification Number
Physical Description
Purity
SynQuest Labs
Not Available
Frozen
TCR-00017-047 Lot #Q7530
White Powder
SE-035 Putified from SE-031 at Pace,Tier I1 08-17-2001
Frozen
W i e Solid 99.9%
LlMS #E00-1715, Page 8 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
TESTSYSTEM
Table 41.Description of test systems used in Solubility Screen Test
I source
Expiration Date Storaae Conditions
I
Millipore Svstem
Not applicable
Ambient
2005 Ambient
2005 Ambient
I PuriWGrade
I ASTM TvDe I I HPLC Grade
I HPLCGrade
I
Table 4b. Description of test system used in Solubility Test Column Elution Method
Source Storage Conditions
Description
Purity/ Grade
Dionex Ambient
I
4x35 mm Hollow Column Body, Frits
PEEK material
Millipore System Ambient
None ASTM Type I
Not available Ambient
Glass Wool Neat
I Source
Storage Conditions
Description
Purity/ Grade
ColePamer
Ambient
MasterFlex US Pump
Equipped with 7
I
casettes Model No. 7524-40
I
I
I
Cole-Pamer Ambient
MasterFlex Microcassette
equipped tubing
Model No. 7519-65
Solva
Ambient
Peristaltic Pump Tubes,
Inner diameter=
I I 1.14 mm
P.v.c C/eltow)
I
I
METHODSUMMARIES
The solubility determination of EtFOSE-OH in water, methanol, and acetone was performed
according to United States Environmental Protection Agency OPPTS 830.7840, "Water Solubility: Column Elution Method; Shake Flask Method" and OECD 105, "Water Solubility" Guidelines, as implemented by 3M Laboratory in methods ETS-8-170.0 "Solubility Screen-Test:Approximate
Solubility Determinationof a Test Substance in Various Solvents; and " ETS-8-171.O "Column Elution Method; Solubility Determinationof a Test Substance in Various Solvents". All analyses were performed according to the method ETS-8-155.0 "Analysis of PerfluorooctaneSulfonate or
Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/Mass Spectrometry".
LlMS #E00-1715, Page 9 of 258
BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
The Solubility Screen Test Method was used to determine the solubility range of EtFOSE-OH in water. The screen test indicated a solubility of EtFOSE-OH in water of less than 10 pglmL at 22.5 "C. Therefore the Column Elution Method was used for the quantitative determination of EtFOSE-OH solubility. Detailed descriptions of the Solubility Screen Test Method and Column Elution Method used in this study are located in Attachment A.
PREPARATOMREYTHODS
0 ETS-8-170.0 "Solubility Screen Test: Approximate Solubility Determination of a Test Substance in Various Solvents." This method is a prerequisite to the shake flask and column elution methods and gives an estimate of the solubility point of the test substance in the solvent of choice.
EtFOSE-OH SE-035 in water: After preparing solutions of water and EtFOSE-OH up to a concentration level that was approximately 5,000 pglmL, there were still particulates visible. Therefore 12.2 mg of EtFOSE-OH were added to 100 mL of water for a concentration of approximately 100 pg/mL. There was still particulate present, therefore 10.1 mg was added to 1 L of water for a theoretical concentration of 10 pg/mL EtFOSEOH. There was still particulate remaining in the flask after vortex mixing, sonicating, and allowing to sit for an extended amount of time. This concentration level indicated that the column elution method should be used to determine the solubility of EtFOSE-OH.
0 EtFOSE-OH SE-035 in methanol: Solvent was added stepwise to a total volume of 1 mL when it was observed that the particulates noticed in the solution may be dust particles. Due to the small volume it was difficult to tell with any degree of accuracy what the particulateswere, therefore a larger volume was prepared for ease of observations. Approximately 170 mg EtFOSE-OH was added to a 4 mL screw top vial and 1.7 mL methanol was added to it. This provided a final concentration of 2 10%. Observations made it clear that the white powder of EtFOSE-OH had gone into solution and was no longer observed. Therefore it was concluded that the solubility of EtFOSE-OH was estimated to be greater than 10% (mass/volume) in methanol.
0 EtFOSE-OH SE-035in acetone: Following addition of 0.1 mL to 10.08 mg EtFOSE-OH,
it was observed that the EtFOSE-OH had dissolved. The dissolution observation marked the end of the solubility screen test and it was concluded that the estimated solubility of EtFOSE-OH in acetone was greater than 10% (mass/volume) in acetone.
Table 5. Summaty of Screen Test method Sample Preparation.
Approximately 10 mg test substance was weighed into tared glass screwcapped vial.
0.1 mL of test solvent (water, methanol or acetone) was added to glass screwcapped vial.
Following the addition of test solvent, the vial was capped, shaken voltexed and then sonicated.
911 1/00 911 1/00 911 1/00
LlMS #EOO-I 715, Page 10 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
4
Followingsonication,the samplewas checkedvisually for undissolved 9/11/oo
particles of the test substance. In acetone, all of the test substance
appeared to be dissolved, and the solubility was then greater than 10%
(massholume) - no additional solubility testing required. 170 mg
EtFOSE-OH was dissolved in 1.7 mL methanol, vortex-mixed, sonicated
and it appeared all substance had dissolved-solubility is greater than
10% at and no further testing is required. A portion of the test substance
remained undissolved in water, therefore further dilutions were
necessary.
5
Up to 2 mL additional water were added and the vial was vortexed and 9/11/00
sonicated. Undissolved particles were still obsetved at this dilution level,
therefore 12.24 mg were added to 100 mL, vortex-mixed, sonicated.
Undissolvedparticlesstill remained,therefore 10.10 mgwere addedto
ILwater, and aftervortex-mixing and sonicatingthe flask,therewere
still particles after 24 hours. Solubilty is< 10 pg/mL and must be determined via the column elution method.
0 ETS-8-171.0 "Column Elution Method: Solubility Determination of a Test Substance in Various Solvents." Approximately 100 mg glass wool was weighed into a tared 15 mL
plastic centrifuge tube. EtFOSE-OHTCR-00017-98 was weighed into a tared weigh boat for a final amount of approximately 100 mg. The EtFOSE-OH was then added to the centrifuge tube containing the glass wool by tapping the weigh boat and scraping particulate into the centrifuge tube. Acetone was then added to the tubes so that the glass wool was completely submerged (approximately 10 mL acetone). Five centrifuge tubes were preparedwith EtFOSE-OH, and two additional solvent blank tubes were prepared with glass wool only. The centrifuge tubes were then placed on a nitrogen blow-down apparatus to evaporate the acetone. The tubes were checked periodically to check for the progressionof the evaporating solvent, and the glass wool was continually pushed below the surface of the solvent so that it was always submerged during the evaporation process. Following the evaporation of solvent, the glass wool was then removed from the centrifuge tube and weighed in a tared weigh boat.
For glass wool with EtFOSE-OH absorbed onto it, the weight of the EtFOSE-OH was
determined by difference of the glass wool weight before and after the absorption process. The glass wool was then put into the 4 mm X 35 mm PEEK column bodies which were taped
vertically to a 1L Nalgene bottle for support. Tubing was affixed to the bottom of the column which was fed through the peristaltic pump and into the bottom of a 15 mL centrifuge tube. The tubing affixed to the bottom of the column was the "inlet" end of the column, where water from the centrifuge tube could flow in. The 15 mL centrifuge tube was filled with
approximately 13 mL Milli-Q" water. The cap to the centrifuge tube had two small holes drilled into it so as to allow passage of the "inlet" and "outlet" tubing to pass in and out of the centrifuge tube. The pump was turned on for each column just enough to fill the barrel with water. After each column was filled with water, the columns were capped and allowed to sit for approximately 2 hours. After allowing the columns to set and equilibrate, the capped top end was replaced with "outlet" tubing which fed into the top of a second EMPTY 15 mL centrifuge tube. The pump was turned on and the columns were flushed for approximately 10 minutes, in which time approximately 8 mL were diverted to secondary centrifuge tubes. The primary centrifuge tube which held the water supply was replenished to a total volume of approximately 13 mL, and the "outlet" tubing was placed into the centrifuge tube just above the surface of the water. The columns with test subtance and method blank columns were
placed in a temperature-controlledincubator set at approximately20 OC. The peristaltic pump was then turned on to allow for recirculation of the water in the centrifuge tubes. The pump
LlMS #E00-1715, Page 11 of 258
BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
flow rate was set and "Time 0" commenced at the time of the peristaltic pump turn on. See
Table 6 documenting the column preparation:
Table 6. Column Elution Method Preparationfor Study (FACT-TCROM), ETFOSE-OHTCR00017-98 at 22 rprn and 11 rpm.
4-Approximately 100mg glass wool was weighed into a tared centrifuge tube.
Sample Preparation- Approximately 100 mg was weighed into a tared weigh boat and then transferred to the centrifuge tube containing glass wool.
3
Method Blank Preparation- started at step number one skipped
step nurnber 2 to step number 4.
4
Approximately 10 mL acetone was added to the centrifuge tubes
and any unsubmerged glass wool was forced below the surface of
- the solvent.
5
The centrifuge tubes were placed on a nitrogen blowdown
apparatus to evaporate the solvent. The tubes were monitored
and as solvent evaporated, the glass wool was continually pushed
below the surface of the acetone to allow for maximal contact with
the solution.
Removed glass wool with care and weighed in a tared weigh boat.
I Calculated weiaht difference from steDl and 6.
7
Filled centrifuge tubes with approximately 13 mL Milli-QTMwater.
8 I Placed glass wool into the emptied column bodies.
9
Fitted tubing to the columns, through the pump and into the
centrifuge tubes. Filled columns with water, capped, and allowed to
equilibrate for approximately 2 hours.
10 Hooked columns up with tubing, put together with the water reservoir (Le. centrifuge tube) and pump. Flushed columns, replenished water reservoir and set up the system to recirculate.
I l a Putthe entire set up in the incubator set to 20 OC. The peristaltic
pump was set to 740 pL/min (22 rpm) to commence flow.
I l b Put the entire set up inthe incubator set to 20 OC. The peristaltic pumpwas set to 175 pUmin (11 rpm) to commenceflow.
7 5 2 7
7
17
I 7
17
7
1O-OW-- 11-07-00
10-0500 11-07-00
104500
t10-0500
11-07-001 11-13-00
11-07-001 11-13-00
*11-13-00
I Io-06-oo I 11-13-oo
mme07 applicable
SAMPLE COLLECTION AND ANALYSIS
At each sampling interval, 100 pL sample was aliquoted in triplicate from the recirculatingsolution
of each of the seven columns into labeled glass autovials containing 500 pL methanol. Vials were capped and shaken to thoroughly mix sample. Autovials were stored at 3-6 OC until analysis in
cooler R-4. On the dates designated in tables 7a and 7b internal standard was added to the autovial prior to analysis via HPLC/MS according to analytical method ETS-8-155.0.
LlMS #EOO-1715, Page 12 of 258
BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
Note: Samples were named according to the time of sample collection and preparation. However, some data points were incorrectly calcuated and the sample ID does not accurately reflect the actual time that the sample was collected. The following table shows the incorrect timepoints as read on the sample preparation worksheets, summaries, and chromatograms, and the "corrected timepoint" is the actual time of collection and preparation (Note not a// timepoints were calculated incorrectly. The table only includes the incorrect timepoints.):
The following table describes the sample collection and preparation regimen:
Table 7a. Sample Collection, Preparation, and Analysis for Study FACT-TCR004 EtFOSE-OH 22 rpm flow
wrate
1
At time 0 hours samples were
7 columns set up to the
10-06-00
placed in a temperature controlled peristaltic pump, 2 blanks and
incubator ID# 1-16 set to 20 `C.
5 containingtest substance.
100 pL aliquots of the column's
supematant (water containing dissolved ETFOSE-OH)were collected and dispensed into
autovials containing 500 pL
methanol.
3 replicates per test system& , per
*a) 1rnL aliquot of supematant was
collected into a 2 mL centrifuge
tube and centrifuged. 100 pL
supematant was then dispensed
into
an
autovial
containing500 pL methanol.
I
I
LlMS #EOO-1715, Page 13 of 258
- t BACK TO MAIN
3M Environmental Laboratory
Report No. FACT-TCR-004
Internal Standard was added to the 600 pL of sample in the autovial.
IS solution information:
Solution "a" = THPFOS oooO1-28-01, 16 pL, 10 pg/mL; and PFBS oooO1-3Cb03,16 pL, 15 pg/mL added
Solution "b"= THPFOS oooO1-21-01 (22.2 pg/mL), 11pL added
I Day 1,28 hours, solution "a"
Day 2,49 hours, solution"a" Day 3,72 hours, solution"a" Day 4,96 hours, solution"a" Day 5, I19 hours, solution "a" Day 6,146 hours, solution "a" Day 7,165 hours, solution "a" Day 7, 166 hours, solution "a" Day 7,167 hours, solution "a" Dav 7.168 hours. solution "a" Day 7,169hours, solution "a" Day 10,238 hours, solution "a" Day 11,264 hours, solution "a" Day 12,288 hours, solution "a" Dav 13.302 hours. solution "a" Day I4,326hours, solution"a" Dav 27.649 hours. solution "b" Day27,649hours,solution "b" (centrifuged sample)
10-1240 10-13-00
10-16-00
~
10-27-00
, a1-02-01
II Day 1,28 hours
Sampleswere analyzedvia
Day 2,49 hours
HPLC/ES/MSon HP 1100 LClMSD D~~3,72 hours
"Hilary."
Day 4,96 hours
I Day5 119 hours
Day 6,146 hours
Day 7,165 hours
1 Day 7,166 hours
Day 7,167 hours
Day 7,168hours
I Day 7,169 hours I Dav 10,238 hours I Day 11,264 hours I Dav 12.288 hours
Day 13,314 hours
Day 14,338 hours
Day 27,649 hours
Day 27,649 hours (centrifuged)
II 10-1740, sequence h001017.s
10-17-00, sequence hOOl017.s
10-19-00, sequence hOOlOl9.s
10-19-00, sequence hOOl019.s
I 10-22-00, sequence hWl022.s
1 10-22-00, sequence hOOl022.s
I 10-2400, sequence h001024.s I 10-2400, sequence h001024.s
10-24-00. sequence h001024.s 10-2400, sequence hOOlO24.s
I 10-24-00,sequence hOO1024.s I 10-2400, sequence h001024.s I 10-2740, sequence h001027.s I 10-2800, sequence h001028.s
10-2840, sequence h001028.s
10-2840, sequence h001028.s
01-0240, sequence h010102.s
01-02-00, sequence h010102.s
Table 7b. Sample Collection, Preparation, and Analysis for Study FACT-TCROW EtFOSE-OH 11 rpm flow rate
LlMS #E00-1715, Page 14 of 258
BACK TO MAIN
3M Environmental Laboratory Report NO.FACT-TCR-004
1
11-13-00
- incubator ID# 1-16 set to 20 OC.
5 containing test substance.
2
100 pl- aliquots of the column's
Day 2,39 hours
11-15-00
supernatant (water containing dissolved ETFOSE-OH)were collected and dispensed into
autovials containing 500 pL
Day 3,63 hours Day 4,87.5 hours Day 4,88.5 hours
11-16-00
methanol.
Dav 4.89.5 hours
I Dav 4.90.5 hours
11-17-OO
I Day 4.91.5 hours
I Day 4,92.5 hours
I Day 4,93 hours
~~
~
Day 5,116 hours
11-lS-00
-replicates per column, per day: Day 6,140 hours
11-1400
Day 7,157 hours
11-20-00
Day 8,181 hours
11-21-00
~
~
Day 9,205 hours
11-22-00
3
Internal Standardwas added to the Day 2,39 hours
600 pL of sample in theautovial.
Day 3,63 hours
11-17-00 11-17-00
IS InfOm\atiOn: THPFOSPFBS ID#O0003-140,16.5pL
Dav 4.87.5 hours
<
.
~ a 4y, 88.5 hours
Da.v 4..89.5 hours
Day 4,90.5 hours
11-17-00 11-17-00 11-17-00 11-17-00
Day 4,915 hours
11-22-00
Day 4,92.5 hours
11-17-00
Day 4,93 hours
11-22-00
Day 5,116 hours
11-20-00
Day 6,140 hours
11-20-00
Day 7,157 hours
11-22-00
Day 8,181 hours
11-22-00
Day 9,205 hours
I1-22-00
Day 2,39 hours
5
Sampleswere analyzedvia
Day 3,63 hours
HPLCESAASOn HP IIO0 LCAASD ' ~ a 4y,87.5 hours
"Hillary."
Day 4,88.5 hours
I Day 4,89.5 hours I Day 4,90.5 hours 1 Day 4,915 hours
1 Dav 4.92.5 hours
I 11-22-00, hOO1122.~
11-22-00, hOOl122.s 11-24-00, hOOll24.s
11-24-00, h001124.s
I 11-2400, hOOl124.s
I
I 11-26-00, hOOl126.s
I
I 11-27-00, hOOl127.s
I
I 11-27-00. hOOl127.s
I
Day 6,140 hours Day 7,157 hours Day 8,181 hours Day 9,205 hours
11-2900, hOO11 2 9 . ~ .
I1-2400, hOOl129.s 12-01-00, h001201.s 12-01-00, hOO1201.s
LlMS #E00-1715, Page 15 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
The pH of each column as well as the temperature of the incubator was recorded at each sampling interval. The pH was consistently approximately 5.5 for both 22 and 11 rpm studies. The temperature range of the apparatus (and therefore water temperature of the sample)
throughout the 22 rpm study was 19.5-21.OOC. The temperature range of the water during the saturation process for the 1Irpm study was 20.0-21.O"C.
ANALYTICAML ETHOD
0 Method presented in Attachment A.
ETS-8-155.0 "Analysis of Perfluorooctane Sulfonate or Other Fluorochemicals in Waste Stream or Water Extracts Using HPLC-Electrospray/MassSpectrometry". Diluted samples (Le. diluted supernatants)were analyzed using HPLC/ES/MSin the negative ion mode. EtFOSE-OH levels were evaluated versus calibration standards ranging in concentration from 2.507-501.4 ng/mL. Internal Standard quantification was used to best fit the data. Target ions were 630 m/z (acetate adduct EtFOSE-OH-test substance), 427 m/z (deprotonated THPFOS-internalstandard), and 299 m/z (deprotonated PFBS-qualitativeanalysis only, not used to report data).
Analytical Equipment
Liquid Chromatograph: Hewlett-Packard@Series 1100 Liquid Chromatograph system
Analytical column: Keystone@Betasil" Cie 2x50mm, 5pm particlesize
Column temperature: Ambient
Cycle Time: I O .0 minutes
Flow rate: 300pUmin
Injection volume: 1OpL
Mobile phase components:
Solvent A: 2.0 mM ammonium acetate in water
Solvent B: Methanol
Solvent Gradient: Time
%B
0.00
15 Yo
0.50
15 %
0.80
60 %
3.50
100 %
6.50
100 %
7.00
15 %
9.75
15 %
10.00
stop
Mass Spectrometer: Hewlett-Packard" Series 1100 API/Mass Spectrometer Detector
Software: HP ChemStation" 8.03
FragmentorVoltage: rdz 630=55 V; m/z 427=100 V; m/z 299= 100 V
.
Capillary Voltage: 3500 V
Gain = 2.0 EMV
Mode: Electrospray Negative
Gas Temperature: 350 OC
Drying Gas: 10.0 L /min.
LlMS #E00-1715, Page 16 of 258
BACK TO MAIN
3M Environmental Laboratory Report NO. FACT-TCR-004
Nebulizer Pressure: 25 psig Analysis Type: Single Ion Monitoring (SIM)
ANALYTICALRESULTS
Data quality objectives outlined in the 3M Environmental Laboratory method were met (see Appendix A).
Regressions. Quadratic curve fit was applied to calibration standards and sample data to
- improve quantitationover the concentration range appropriate to the data. All calibration
curves had least-squarefits of 0.990 or greater (R2values ranged from 0.9913 0.9998 for 22 rpm flow, and 0.9971-0.9993 for 11 rpm flow).
Calibration Standards.All calibrationcriteria were met. It was not necessary to use
exponential or quadratic fits of the data, as the concentration range was linear. The curves
were examined closely, particularly for accuracy of quantitation at the low and high ends of the curve. It was necessary to use an abbreviated portion of the curve to better fit the data range and provide better accuracy for the data range quantified. The calibration range used for the majority of samples to quantify the data was 5.01 ng/mL to 401.Ing/rnL, exceptions to this calibration range are listed below in table 8:
Table 8. Calibrationcurve ranges usedfor the quantificationof EtFOSE-OH 11and 22 rprn data.
I h1017a2.b
h1017bl .b
h 10 1715. b
I 2.5250.7 ng/mL
2.5401.1 ng/mL
5.01-250.7 ng/mL
I
22 rpm study samples
h i024& b
10-401.1 ng/rnL
hi122bl.b
5.01-501.4 nolrnL
~- ~~~~
11 mrn studv sarnDles
~-
0 Limit of Quantitation(LOQ). The LOQ is equal to the lowest standard in the calibration
curve. The LOQ for the majority of sample batches was 5.01 ng EtFOSE-OH /mL, with the exceptions to this in table 8 (in bold print) above.
Sample Replicates. Sample replicates must meet the 30% precision criteria to be reported. Samples were excluded by one of two scenarios: 1) suspected outlier value was excluded via Q-Test (statistical analysis) or 2) the %RSD for the measurement was greater than 30%, therefore signifying that the data was too scattered to draw a conclusion. It was necessary to exclude the following samples to meet the data quality objective of 30% or less precision:
0 22 rpm data. Samples 100600-CEM-EtF-146.2.1,100600-CEM-EtF-146.2.2, 100600CEM-EtF-146.2.3 (Day 6 (146 hrs), column 2); 100600-CEM-EtF-146.4.3;100600-CEMEtF-167.1.I;100600-CEM-EtF-167.3.3; 100600-CEM-EtF-167.5.3; 100600-CEM-EtF238.3.3, 100600-CEM-EtF-238.4.2; 100600-CEM-EtF-264.2.3; and 100600-CEM-EtF288.5.3 were excluded from the reported data using Dixon's Q-Test with greater than 80% confidence. Samples 100600-CEM-EtF-166.5.1, 100600-CEM-EtF-166.5.2, 100600-CEM-EtF-166.5.3(Day 7 (167 hrs), Column 5); 100600-CEM-EtF-302.all (all samples for Day 13); and 100600-CEM-EtF-326.alI (all samples for Day 14) were
LlMS #EOO-i715, Page 17 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
excluded because the data was too scattered and did not meet data quality objectives for precision (<30% RSD).
11 rpm data. Samples 110700-CEM-EtF-63.4.1, 110700-CEM-EtF-87.4.2, 110700-
CEM-EtF-88.5.2, 110700-CEM-EtF-89.2.alI (column 2, repsl,2,and 3), 110700-CEM-EtF89.4.3, 110700-CEM-EtF-90.5.3, 110700-CEM-EtF-92.5.1,and 110700-CEM-EtF-116.5.2 were excluded from the reported data using Dixon's Q-Test with greater than 80% confidence. Samples 110700-CEM-EtF-87.all (all samples for Day 4, 87hrs); 110700CEM-EtF-90.4.1, 110700-CEM-EtF-90.4.2, and 110700-CEM-EtF-90.4.3(Day 4 (90.5 hrs), Column 4); were excluded because the data was too scattered and did not meet data quality objectives for precision (<30% RSD).
0 Continuing Calibration Verification. For quantitative determinations, a mid-level matrix calibration check was analyzed every five samples to monitor instrumental drift, with a limit of +5I% of the target concentrations.
Blanks.
Method blanks (Milli-Q water taken through the entire sample preparation, dilution, and analysis process) provided a measure of laboratory contamination. Acceptable values for the blanks were less than 50% of the limit of quantitation (LOQ).
0 Solvent blanks (aliquots of methanol) provided a measure of reagent and instrument contamination. Acceptable values for the blanks were less than 50% of the limit of quantitation (LOQ).
Specificity (according to OPPTS 830.7840): The solubility determinationas stated in the EPA Guidelines follows. This method should only be applied to:
0 Pure substance. The official purity is to be determined, however comparison results between SE-035 (purity of 99.9%) and TCR-0017-98 from this study indicate that within the limits of experimental error, there is no distinct difference between the two lots. Therefore, for the purposes of this study, the purity of TCR-00017-98 is 99.9%.
0 Substances that are stable in water. Hydrolytic studies have been conducted at the 3M Environmental Laboratory showing stability.
DATASUMMARY AND DISCUSSION
Tables 9a and 9b summarizes individual sample data. Representativechromatogramsare
-
presented in Attachment C. The table displays EtFOSE-OH concentrations for individual
injections for the 22 and 11 rpm studies. Also included are the average concentrations for each
column triplicate, standard deviation, and % relative standard deviation. Sample calculations may
be found in Attachment E.
For the 22 rpm study, samples were collected once daily up through day 6. It was postulated that the solution had reached a plateau in concentration by day 6, and the multiple sampling (i.e. collection of 5 or more consecutive sample sets) was therefore performed on Day 7. Samples were continually taken once daily after Day 7 until analytical confirmation could be made that the solubility point had been reached by Day 7. By evaluating the data, results indicate that the
solution became saturated by Day 2, and the 5 consecutive timepoints on day 7 confirmed that
the solubility of EtFOSE-OH had been reached, and could be reported within 30% variability as required.
LlMS #E00-1715, Page 18 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
By evaluating the data presented in table 9a, it may be noticed that for timepoints 238, 264, 288,
302, and 326 hours the concentrationis almost double that of the rest of the data. It was
discovered that this disparity in concentrationswas due to a sample preparationerror. These
timepoints were therefore plotted as diamonds to denote outliers of the saturation trend. The 96
hour timepoint is also plotted as a diamond symbol because it is also an outlier to the saturation trend. By averaging evaluating all data points prior to 238 hours plus the 649 hour timepoint, the 96 hour timepoint may be excluded via Dixon's Q-Test.
Table 9a. EtFOSE-OHSample Concentrationsby timepointfor 22 rpm study.
.. I
I Day 2 (49 Hours)
Da-v 3 (,72 Hours) I
Day 4 (96 Hours)
Day 5 (119 Hours) Day 6 (146 Hours) Day 7 (165 Hours) Day 7 (166 Hours)
Day 7 (167 Hours)
Day 7 (168 Hours) Day 7 (169 Hours) Day 10 (238 Hours) Day 11 (264 Hours)
Day 12 (288 Hours) Day 13 (302 Hours) Day 14 (326 Hours)
F Day 21 (649 Hours) j
151 155 209 133 150 179 152 157 155 154 340 296 359
:;:499
--,
14,9% 25. 16%
53, 25%
11,8% 24, 16%
35, 20%
21, 14% 43, 27% 28, 18%
30,19%
61, 18% 74, 25%
89, 25%
314, 63% 139, 37% 31, 18%
5 columns. 3 replicates per column= 15 samples 5 columns. 3 replicatesper column= 15 samples
I
5 columns, 3 replicatesper column= 15 samples
1 5 columns, 3 replicatesper column= 15samples
3 columns, 3 reps per column+ Icolumn, 2 repsper column=lI samples
5 columns, 3 replicatesper column= 15 samples 4 columns, 3 replicatesper column= 12samples 3 columns. 3 replicates per column + 2 col, 2 reps per column= 13 samples 5 columns, 3 replicatespercolumn= 15 samples 5 columns, 3 replicates per column= 15 samples
3 col, 3 replicates per column + 2 col, 2 reps per coI= 13 samples 4 columns, 3 replicates per column + 1 col, 2 reps per coI= 14 samples
3 columns. 3 replicates per column + 1 column 2 reps per coI= 11 samples
Dataexcludeddue to pooroverall precision...data too scattered. Data excluded due to poor overall precision...data too scattered.
5 columns, 3 replicates per column= 15 samples
LlMS #E00-1715, Page 19 of 258
_BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
7
I 1
FACT-TCR004, EtFOSE-OH Solubility, 22 rprn Data Summary
400
350
300
250
200
150
100
50
0
0
100
200
300
400
500
600
70C
Hours
Since the saturation point of the water solution occurred after only 2 days in the 22 rpm study, the multiple sampling point for the 11 rpm study was performed at the 4 day point, to ensure that the solution had sufficient time to saturate. On the fourth day of the 11 rpm experiment, six consecutive samples were collected, and then samples were also collected for four more days after to document the behavior of the solution. As the results indicate, saturation did occur by day four for the 11 rpm study, and the final data reported for the 11 rpm solubility point was based on the average of day four's results.
Table 9b. EffOSE-OHsample concentrations by timepoint for 11 tpm study.
Highlighted samples denotti excluded or unacceptable data.
LlMS #E00-1715, Page 20 Of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
FACT-TCR004, EtFOSE-OH Solubility, 11 rpm Data
Summary
,
50
100
150
200
25(
Hours
The final solubility of EtFOSE-OH TCR-00017-98 in water and EtFOSE-OH SE-035 in methanol and acetone is summarized in tables 10a and lob:
Table loa. Summary Table of Solubility of EffOSE-OHinWater.
11 rpmstudy Final Value
I I I 159
10,6%
~
~~~~
5 timepoints
~
142
6,4%
5 timepoints
151
11% RPD
2 consecutive runs
The solubility of ETFOSE-OH TCR-00017-98 in water was determined to be 151 ng/rnL (corrected
for purity) 19.5-21OC.
Table lob. Summary Table of Solubility of EffOSE-OHSE-035 in Methanoland Acetone.
Methanol Acetone
> 10% > 10%
The solubility of EtFOSE-OH SE-035 was determined to be greater than 10% (mass/volume) in methanol and acetone at 22.5 OC.
Attachment B contains data summary tables.
LlMS #E00-1715, Page 21 of 258
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
STATISTICAMLETHODS
Statistical methods were limited to calculating means and standard deviations. Data that did not meet acceptance criteria as described in the OPPTS and OECD guidelines was excluded
following statistical examination using Dixon's Q-test. Refer to Attachment E for applicable
equations.
STATEMENT OF CONCLUSION Under the conditions of the present study, the solubility of EtFOSE-OH TCR-00017-98 in ASTM Type I water is 151 nglmL 19.5-21 "C. The solubility of EtFOSE-OH SE-035 was determined to be greater than 10% (masslvolume) in methanol and acetone 22.5 "C.
REFERENCES
1. Fate, Transport and Transformation Test Guidelines Office of Prevention, Pesticides and Toxic Substances (OPPTS) 830.7840 Water Solubility: Column Elution Method; Shake Flask Method. EPA 712-C-96-041, August 1996.
2. OECD 105: Water Solubility. Adopted 27July, 1995. 3. Harris, Daniel C. Statistics. QuantitativeChemical Analysis. 4th ed.; W.H. Freemanand
Company: New York, 1982; p 70-71.
4. Natrella, Mary Gibbons. The Treatment of Outliers. ExperimentalStatistics, National Bureau
of Standards Handbook 91; U.S. Government Printing Office: Washington, D.C., 1963; Pages 17-3, and T-27.
LISTOF ATTACHMENTS
0 Attachment A: Extraction and Analytical Methods 0 Attachment B: Data Summary Tables
Attachment C: Sample Chromatograms 0 Attachment D: Deviations from the Protocol 0 Attachment E: Sample Calculations
LlMS #E00-1715, Page 22 of 258
SIGNATURPEAGE
BACK TO MAIN
3M Environmental Laboratory Report No. FACT-TCR-004
We certify that this reportis a true and complete representationof the data for this phase of the study:
Mark E. Ellefson Principal Investigator
0&s5/7
William K. Reagen Laboratory Manager
Date
UJ7//0/
Date
LIMS #E00-1715,Page 23 of 258