Document 1gjoE8nEqw8eQrN0X0mNadzGK
AR 226 - 1204
CQonPte DCometCfoCn SAeSacd Seences
Analytical Report
Willan. Raiford
L Wilion Bucton ~~
CRT
CRP 713
nBaarbCarriasStinaLaPrisveenn CER2P28702
LMaormiaH.nnLeeuMnagrsi 3scArlloennzTearnnen,
E3a203z TaRnCe E1340/11266
DATE: 16 January 2003
E3 R2.3 sc 38
v=
E2 55 a
Ly
TO: George Senkler SITE: Fluoroproducts, Chestnut Run 711
FROM: Mary A. Kaiser and Michael J. Michalczyk
--_--
Determination of Perfluorooctansic Acid from the Surface of Commercial
Frying Pans
Abstract
A method
was
developed
to
determine
if
perfluorooctanoic
acid
(PFOA),
a
sfluurofraocpeoolfymceormmpeorlycmiearlizfartyiinogn
aid,
pans
mciogahttebdewiptrhesaenftluoinroapnodlyemxetrr.a
ct
A
ed from the
modified US
FDA method was used for extraction and an LC/MS/MS method used for identification and quantitative analysis. No PFOA was found in the water extract
from the fluoropolymer surface to the limit of quantitation in water, 50 ng/L (ppt).
from two 500-mL portions of water.
Request
"A
request
wos
made
to
determine
if
any
quantifiable
PFOA
might
be
found
in
extract water from a fluoropolymer surface. A US Food and Drug Administration
t(hFeDAf)rymientghpoand. was modified fo extract PFOA, if it were present, on the surface of
Fluoropolymer dispersions are used in the commercial processes for coating
metal. These products frequently use a fluoropolymer polymerization aid, PFOA,
during their manufacture. Recent studies' have suggested that some
UPIND The miracles of science
Page 1
fluoropolymer polymerization aids might persist in the environment and might
elxevheilbsitofsothmeesteoxmiactoelroigailcsalmiefgfhetcbtes
in animals.
present in
hAunmoatnhebrloroedpsoerrt?umi.ndSiicnacteedPtFhOaAt
low
is
unseecdesfsoarsyusfpoednedtaenrdmienmeulisfiPfyFOsAomies sptoilllypmreerssednutraifngtetrhetihremcaonautfiancgtaunrde,baitkiinsg
processes and thus might be a vehicle for human exposure.
Experimental Section Ten frying pans,
five
8-inch
Farberware
Elite
Cusine
uncoated
stainless
sotfepelolayntdetfriavfeluBo-rioncehthAynloenleon(PTCFlEas)siacndhaPrFdA-a(nbordainzideedd aalsuDmuiPnounmtcoAautteodgrwaipthha)bwleerned
pmoudricfhiaesdedasfrnootmead. loTcahlebpeadnsanwderbaetfhirssptecwiaaslthyedstaorse.recAonmmFeDnAdepdrobcyedtuhree' was
mthaennufraicntsuerdewritwhitthharpeper5o0ximmaLteolfyp5or0timoLnsofof0.r5in%se
Palmolive detergent solution (deioinzed) water and dried
and with
a Sonora towel. A custom-made glass #19 female ground glass joint attached
lid in
(9" the
diameter center of
x 0.125" thickness) with the glass (see Figure 1)
a
and a water
condenser and dried.
were The
washed pan was
with the detergent placed on a Vulcan
solution, rinsed with model 38L gas stove
deionized and deionized
water was added fo and condenser were
approximately placed on the
0.635 cm (0.25 inches) from the pan, and two 201-g weights were
top. The added fo
lid the
lid
twoateenrsuarnedacognodoednsseealr. waTshheeactoenddeunnsteilrtwhaeswcaotoelredrewfiltuhxweedtcoicnet.inuTohueslfyryiinnsgidpeanthweith
ccoonntdiennuseerfo(rFi3g0urmein2u).tesT.heThheeanttwhaesatpupranreadtudsowwansarnedmroevfelduxfwraosm atlhleowbeudrnteor and
allowed to The
cool cooled
water
was
then
placed
in
each
of
two
500-mL
wide-mouth
polypropylene bottles. and the pan was rinsed
The with
lid and three
condenser were rinsed with 50-mL portions of deionized
deionized water, water. All rinse
water wTahse apdrdoecdedfuoreeawcahsorfetpheeattewdofpoorlytphreorpyelmeanienbdoetrloefs.the frying pans. When
testing had completed, the within an insulated cooler.
polypropylene bottles Ice was placed on top
were placed inside of the bottles and
a plastic the liner
liner
folded and taped custody tape and
fSocoptrcehve8n9t8l1eatkapaeg.e.ThTehecocooloelrerwawsassetnhtenfoseEaxlyegdewniRtehscehaaricnh-of-
(rSetcaeitveeCdololnegiec,e PafA)EvxiyagAeinrRbeorsneearEcxhprreecsesivfionrg,oveexranmiignhetd,delloivgegreyd. inA,llansdamsptloersedweirne.
locked
storage Samples
until were
transfer treated
to the Exygen Research analyst. according to Exygen Research method
01M-008-046
entitled "Method of Analysis for Determination of Ammonium Perfluorooctanoate
Page 2 POND The miracles of science:
5
(mAePtFhoOd),iwnaWtaetrers"amipslseusedweorneJtarneuaatreyd1w1,it2h00220.04(SLeoefAspopdeinudmixthLi)osuUlsfiantge tshoilsution, smoeltihda-npohlasien ewxattrear,cttehdenwietlhutaeCd-w1i8thre1v0er0s%e-mpehtahsaenocla,rtarniddgteh,eellauttteedr wpiotrhti4on0% collected for analysis by LC/MS/MS. The treatment results in an eight-fold concentration prior fo analysis.
Results and Discussion "All water from the
fluoropolymer-coated
frying
pans
showed
no
quantifiable
PdeFtOeActiporens(enLtO.D)Woifthapaprloimxiitmaofteqluyan1t0itpaptiopner(LalOiQqu)oto,f
50 this
ppt and a limit of number corresponds
to
100 ng/420.42 cm? LO or 20 ng/42042 cm? LOD.
result
One frying pan with no coating (Table T) in one instance and a
(just a stainless nondetect (with
steel surface) gave a positive an approximate 10 pp limit
oafnaldyettieccatlioanp,pa1rLatwuast,eers)peicniaanlloythfelruoirnosptoalncyem.eriScinpcaertPs,FObAackisgeraosuinldy aledvseolrsbceadnobneo
problematic. PFOA can be Chromatographic systems.
found in analytical solvents The background levels are
as well usually
os transfer lines in around 10 ppt. The
UfiSveEtPiAmeGsuitdheelibnlea*nkrelcevoemlmsehnodusldtbhaetraenpyormteeadsausreumnednettsecotbetdai(nUe)dbtehcaatusaerethleess than
quantity reported background. Due
cannot reliably to background
lbeevedlissacnrdimtihneatpeodssbiebitlwietyenfotrhecasrarmypolveearnodr
the
loabbsoerravteodrythcartostsh-ecobnatcakmginraotuinodn,letvhele, lciamritryofovqeura,ntoirfliacabtoiroantowrayscsoenttaamfin5a0tippotn. We
sometimes exceeds the 50-ppt level.
References 1. Guide fo
the
Safe
Handling
of
Fluoropolymer
Dispersions,
The
Society
of
the
2.
Plastics Industry, Inc. 2001p. KJ. Hansen, L. A. Clemen, M. E.
7. Ellefson,
and
H.
O.
Johnson,
Environmental
3.
Science US FDA
aPnrodcTeedcuhrneo,losgeye,r3e5f,er7e6n6c-e77201C(F2R01071)5..300(d)
fable2
condition
of
use
8.
4 "PDraotgaraVmalRioduattiionne SAtnaalnydtaircdalOSpeerrvaitciensg",PUroSceEdPuAreRefgoironCoIn,traActtheLnasb,orGeaatrogriya,
Revision 2.1, July 1997.
Notebook Numbers 104373-1 10 -15.
@UPOND The miracles of science"
Page 3
Control Terms to be included PFOA, APFO, frying pans, extraction, LC/MS/MS, US FDA
sampleTable I AnalPyantiIc>al Results 33776666003312 SS5e#t1., 12"aaliiqquuoott 76606-15542, 1" liquot 333777666666000290112 S SS5#S 53, 2# #21%%aal2 3lliiiqqq, u,uueottt 6Sr6e112221 sSse, 12%aailqiuquoott 3S"r7s76666161515.612 SSOL5H#Sw5,a,te12r7%caaollniitqqruuoooltt 61100003305 5555 ##11. I2sntdlliqquor 1100006605 CCoooatteedd ##11,, I2sntdolailgiquueott 110000995 CCooaatteedd ##22,. I2sntdaclliiqquuoatt
"4100112205 CCoooatteedd ##33., I2sntddailqiuqouott 6G1l00115306 5555 ##22,, 12nsdldiqiuootr 61111003305 5555 ##33,, I2sntddlqiquuoor 46115006300 C5o5at#e4d,#I4st,dIistgoaltiquot 6C1l50o3t5 Scosst#e4d, #25n,d dsitquaotir i41e5t09s0 C5o5at#e5,d #I5st,aolridquooltiuot 6G1i5t0z9o5 5c5om#e5,d#2nidsatliaqiuogtur GGiiss11z5s0 C5o5ot#eLd #Is6t,anlidquoatliquot Gi1s5i1s650 O5F5 w#a2t,er2ncdonatlrioqluot 411160 DLT0wQa=te5r0cponptt=rolS0ng/L
Lob+ c107pt
PFOA Peak area
(ng/L)
nd
eri7g9i2nal
3wd9 s71a
n9ndd0 9e680z90
nwdd a5183
nnnddd
a4215
n0d 13o7030
nndd 1258984
11dd 8101064
nndd 10a3it9
nnee a5687
2W3o asmeees
nwdo 3s0e9ss
w nd ma 39s
nwdo 2a5s3e8
nend 426120
1Wdo warm
a21m8s4
ndn=e rat 1658
detected
Peck aren
lab r2ep6licate 570468 6m5895e%6 a738 356358458 11307794 17593 1825306 1p0e36 750424 3o95s6 3s9214 232m4 209928 24356009 33614498 2200502 7s
Page 4 @UPOND The miracles of science s
a in Figure 1 Glass Lid for Frying Pan
i
Be
2
o--
GUPIND The miracles of science:
Page 5
4
Figure 2 Experimental Set-up
a Jo ae
a Fo
L a e
EL al = = PREEEEE
ae
ii
:
Ll
BE
x
=7
2
CE
Appendix 1, Exygen APFO Method
TE
Method of Analysis for the Determination of Ammonium Perfluorooctanoate (APFO) in Water
AUTHORS
ohn Flaherty and Karen Risha
DATEISSUED
January 10, 2002
Page
@UPIND The miractes of science"
7
sponsor
Dupont Corporation
PO BOX 80302 Wilmington, DE 19880-0302
PERFORMING LABORATORY Exygen Research
3058 Research Drive
State College, PA 16801
METHOD NUMBER 01M-008-046
Page 7
GUPIND The miracles of science:
g
MANAGEMENT APPROVAL
John Flaherty
Date
Laboratory Manager
Exygen Research
PRriecshiadredntA. Grazzini, Ph.D.
Date
Exygen Research
Mary A. Kaiser, Ph.D.
Date
Sponsor Representative.
Dupont Corporation
TABLE OF CONTENTS
BL CHEMICALS wns MANAGEMENT APPROVAL .............. wesersone rss sansa
TABLE OF CONTENTS... -- -- assesses
LIST OF FIGURES...
--------
seniors cere arses
1. SUMMARY ......... escent
eter wosaersrrs roensn
2. EXPERIMENTAL COMPOUNDS............. erent rss
3. CHEMICALS AND SUPPLIES rr
-- nse
essnssseseen arse nesses ans nse ssssns esses anaes
3.2. STANDARDS S--_--
esssssss sree
srs sss
3.3. EQUIPMENT AND SUPPLIES......c.ovrvsvrorrsrree evseressrsss aera
34. SOLUTIONS...
-- -- sae
verse sems aasees
3.5. PREPARATION OF STANDARDS AND FORTIFICATION SOLUTIONS......c0oovn
35.1. Stock Solution....... rsrren
assess reser sess
3.5.2. Fortification Solutions essere sree nan esses ssesesssaesssns
35.3. Calibration Standards. rseerson rss ssn
354. Controlling Standard Bids................ conan
essere
4. METHOD...
--
-- ---- rss
nn
Page 8
WAND The miracles of science:
q
--
AL FLOW DIAGRAM...
4.2. SAMPLE PROCESSING. -- rope _
ES
--
4.3. SAMPLE PREPARATION.........
ns
toe
-----
wersesseeressanan
48 EXTRACTION. css 45.
QUANTITATION.
45.1. LC/MS/MS
_-
System
esses
and Operating Conditions
cwerestrsrressseeeesss
(Electrospray).
452. Example MS Operating Parameters...
4.5.3. Calibration Curve Procedures...... -- essstssreess sae wo
46. ACCEPTANCE CRITERIA... 4.54. Sample Analysis........ _
--
on ore ono
4.7. MASS CALIBRATIONCRITERIA .......
eee sss
4.8. PERFORMANCE CRITERIA. -- crop
esses
4.9. METHOD DETECTION LIMIT DETERMINATION. ........ esses
4.10. TIME REQUIRED FOR ANALYSIS ..............
ers
5. CALCULATIONS.......... essere: wercens
evssssaeress
ans
sass
eeer eases
ssa
ra
seeseses
7. REFERENCES... 1. SUMMARY
remssnsns sms
-
EE
------
T(hAiPFr)epianrtwadteert.ails a method of analysis for residues of ammonium perflusraoctaneate
APFO forms perfluorooctancic acid (PFOA) in solution. PFOA is extracted from water
uaussciicnnoggmpCslyeislseshcoetldeiddbrypehalacisqteuiiodenxctmhroranocimttaiotoronign(rgSaPp(ESh)RyM!c)/a.tratrnidRdeegsmeismd.uaesssQuosafpntePicFftiOrcAoamtefitoorunynod(faLPrCeF/OMmASat/hiMseSm)ataincaallylsiys ocfontvheirstemdetahnodd.reported os APFO. The chemical formula of APRO is given in Section 3 Tm5he0etnhgpo/rdLopw(ahasirectdhs-lgpiievrte-stoarfidleqlquoaunna)ttietfaortreicoAonPvO(er.LyOa:cctohrdeilnogwetsotEfPoArtgiufiidcaetliionness)pfeocriftiheidsbmyetthheo is QWhuaeatneterixftairncadacttpiirooonnciepsrospceeedrdfutroherr,moeudaglhlustsiahnmegpelcxeatslriab(crianttciliouondnipnsrgtoacsnetddaaunrrddeasr,dpsir,deepnctoainrcteardlolfisno, fsfialomtrpeilrfeeisd.catt|yipouenrsTienrgs.) aTirnhejeeccmtoeentdchedonudtrridanetgteetdchtebiyocnhrlofimamaictttoo(rgMrDoafLp)heiiicgshrtau:pnptrihosexrieeqmfuaoitrveeal,lyent6theffoosm12a50l0lnesgntg./sLdtea(pncedonandcriednng(t2ro5anterhsafe6t).0 i1n3s6trAupmpeenntduisxe8d. for analysis. This value is based on procedures found in 40 CF Part
2
Experimental compounds
Page 9
@UPOND
The miracies of science
10
The structure of ammonium perfluorooctanaate i given below.
CF3(CF2)sCO0" NHsTM
CFheomical Nome = Molecarweight =
4A3m1monium Perfloraoctanoate
celFeF ieFleo
PrF eLtFe
0" NH,"
aNsostaey:. The PFOA (anion) (formula weight 413) s present inslution and i used for the
33_7__cCHhEeMmiIcCaAlLsS ANDsuppLTES
Chemical Methanol (720M)
Grade Hoc
Sores Thaker
Coraing IT50932
WAamtmeornium Acetate RTeygpeenTt Sigimna-hAoludsreich A5:30
TSyopdeiuLmwTohtioesu=lfeloetcetrical
resiRsetaivgietny,t mnimam IoTf
baker 16.67 Wi/am
a7
. 25C.
from
a
2.2 StonderLdasbcanco Waterpro workstation
Standard
Lot Number purity (8) Source
Paecndtadecafliorooctanoc 0831600 96 Arch chem
Perfivorcactanoic Acid 14D 9 Florochem Us
3 Eaipment ondSuppe ly ee ------------eemerm----re
Balance, analytical (Esqpuilpamenlteast 000073)
Fr Mettler
VViasciupuremppvuamcpium manifold
SuBpueclhco
(pSoerptPo#kWVAacTb0c3c6(715g5))1C16 cartridges
Waters
5150 mmLi cdiissppoassaabbll ppooblpyrprooppyylleennee cceennttrriiffuuggee tfuubbeess
wwwee
Disposableglassmicropipets (50-1004. 100-200 Drummond (VWR)
2w5ymm diameter glass fibre acrodise 0.45 1 (cat
Gelman
Page 10
QUPOND The miracles of science
1
sGlaoss2s3A)pets nd volumetric flasks
various suppliers
(HSpytoparentrd#c-ac8robn4e4d0rdo1rpo7-p4i-0ni0ngu)gauradrdcoclrutmrnd(a4emhml)der
Keystone.
Keystone
H1P2L5CmpuLmOpP(EnLCaIrOrAoDw)mouth boties 2 mL clear HPLC val it ca# 561.3400)
SNhailngoednzeu He
Standard lab equipment (graded cylinders, rious suppliers
dLiCs/poMsSa/blMeStaunbdesHePtLcC) ystems -_
e4c5sedicetsimcornsi4bes5d in
Ncoatneb:e sEhqouwinlteontprmoadtuecreiaslasfimsafayctboerysurbesstitteuted fo these specified in this mth if hey
N1oteIsn order to ovoid contamination, (rues, piper, ete)
the
use
of
disposable
obware
is
highly
recommended
2. PHTPFLE aourtoPsTaFmEplelirmneudsctonnattainbeers05or. equipment, including PTFE-ined HPLC vil fo the
3. @rTeyHfpyeeprrIeerwdca1at0erraqsuus"afeiddltdceuorrleiudnmgtnytpuheseiTnsgwaamaptlrHe"P,LaCnhdeprusmetpaa,nfdtoaserrddenesxcStrrhiasbcetrdieopmnorsstheocutlidonbe4.2f.iltTehriesdwtahrreonugh
4.
5.
ULTtsCe/isMdSnies/pcMoesSsasbbalereyfomtrioecrucoshepe.icpkeCtteshtearisopllivopetenntsusmT(bomeeartlshiaqhnuaoevle)sbtfeaoenrndatfrhodeunspdarue1rsieobnencsuen1osoftmcaaobnklteeamcfidonorarnvtaosi.boyn
standards and sample fortifications
3.4. Solutions
S35o:0l8u6tionmofMbyamamofonanicmotemonaoicfmet2a5tetcoaenmdradkeeistshiesno2gutmiinMo1namomfiastryipuempLrwaeocpteaetrrae.tde Dsailbktyeiotnhwewee5ei6dghmfionirg mobile phase A
(2) 40%wimtehthfainor!edis ympadee bwyatmeerasuring 400 i. methanol ond adjusting the volume 10 11
(3) 250pmeg/mTLuastoedrium thiosulfate solution is made by dissoing 25 g in 100 mi. filtered
oNroetpea:reTdhaesalfoonrgeamsentthieornaedtiexooafsmphleessoavrenptro1voidseadutfeoraqtuiiodsanocnee, malatmemranantciev,e volumes my be
3.5. Preparation of StandardsandFortification Solutions
Anciytical standards ar use for hree porposes
Page 11
QUPIND The miractes of science:
12
1. 2.
CaLaarlbeioburrsaaettdoiroony ScCtoaanlnridobalrrdeSspti-hkeTehrseess-peoTsnhtseaesneodfafrhodrestiafdieectaeptcirtoeonpsraruaersdeeipnrfeitplhatereeradednoTyryapceoncewnattreartiaonnsd
5. MrceoacrtorrveiesxrpyoS.npdiLikanebgso-rtaoTthoterhsyeecLfoonOrttQoilfiascnpaditikoe1ns0sxaarLerpOprreepapanardreedadrmbeyfusispeeidnr1g0Tiydnpetethemreemrifncieeedamsaamyphleees
uotsecdonToceenltruaattioensheranegfifnegctforfomth5e0s0am1p0l/eLmuaptr1i0x 5o0n0o,n0l0y0tncga/LrecoMvaerryi.n spikes aes
sTrhaepoarbtsioolnusteofvoslluamtees1o0fstoheensttaanedarmadisntmaaiynedb.e varied by he arly as longa the caret
vSPotrlaeunpmdaeartrerdiac(scftoloar3scrkk5.e.cs1to.eTludhteSifotosnrotoocpfckukr1is0stoo0yll)uuutta/iinonodnnL(diionlfu1tP2e5F-fOmoiA.1bL0y0OPwmeEiLgbhowiitntighesom)uet8th1a03nmeg)beomfsotaonrdoeypdteinmcsas.l aretferigoefrparteopraratat2ioCn fo 6C and is stable for a maximum period of 6 months from the
a.
1Pr0epmalreof3.tf5oh.re2t.i1f0i0cFaoptrigto/niLfs.itcasanttdoiacokrndsooSlfoult1iuo0tniuo1g0n/.sLa.10(01-0m0L0. wp 10 volume with methanol
nvo/lmum)etroifc
PlFaOkA
obynacriinrge
5. hPreepa1r0e.g/fmoLtisfoclauttiioonntsot1an0d0amrkd.owfit0h.1mget/hman.ol(1i0n0anvgl/ummLe)tbnyciltiang 100 mL of
EC.xampPhlreee:p0a.rt1ewgoa/hfmouLrntdisrfoielcdtaitmoiioncnr1so0lt1iat0ne0drasmrLdo.fowfitth0h.e0m11e0tgh/nagmn/o.lmL(n1s0canusgt/iemoLhn)esbpyneiditcithnog 1400.0mmiL.. ooff
water is equivalent to a 0.05 ppb (50 ng/L) fortification.
waSthtio2crheil1ml0ef6oiCrttiiffoincreaacteimosensxasirtynaunfmdoapmreadrkiseoodlnuoetfiwo3nsstmaionnndatahrrsdesffrurisoigmnegrsahttehred(saitneoc1ok2f5o-ptrmieLopnLar.OaPtENioobntoteotllfeesrn) ha additonal concentrations my be prepared f necessoy
PLyEp/eMrSca/rMbS_dcra3ol5pb.ir3nat.igouCnaarsldtiobnrcdaaorttrirdoisdngaeSr.teapnrTdehapearrdecsdaliinbrtatyipoen through the extraction procedure. identical To samples.
Tswtaantdearrdis.taevde The extracted
ptrhorcoeusgshesa rancoree
may be usd for period of wo weeks when sored refrigerated a 3 10 0)
nTeheedefdollwing is o typical example: additonal concentrations may be prepared os
Page 12
QUPIRD he miracles of science:
13
Concentration Fortificati
of Fortification on Volume
(Snogl/utmiLo)n
wt)
Volume of Fortified
(m1) Control Sample
Final
Concentration of Calibration
St(anngd/aLr)d
Calibration
Standard ID (example)
0
0
40
0
Xcmmddyy-0
10
10
100
200
40
40
25
Xemmddyy-1
50
Xemmddyy-2
10100
140000
"400
215000
XXecmmmmddddyyyy--43
110000
02000
a400
1500000 XXcemmmmddddyyyy--65
* ecTxohntecreaencxttrteartdaiccotnae,ldbdrcouotenicooenntstrthaaentdicaoornndc.oefnttrhaeticoanliobfratthioenssttaannddaarrddrisi. enqguatlhteoe8xxtriatcstiinointia(lSPE), XC =
spArteazpneadrraoerdtsshteeaxnstdtraaarncddtaerdsd.oslutaTinhodnisi(snrtoeaatngdieannrtcdlbuilsdorueakssd)edpmaurostnltoyffbtoeheaoccrcaeetpsbasrraestdhieownirctehaugeecnatcsh sseeetd oouf
S6toCr,eupaltlo ewxotrwaecetekds cafbation standards in 15-mL. polypropylene tubes at 2C fo
35.4
CPorneptarroellaingsepSatraantdeasrtdacBkiassolution Standard from a different source
boyf
w1e0ig0hign/gmoLut
f1r0omm
ga
odfifafnedryetnitcaPlFsOtAanndenadt
(voclourmreetcrtiecd flfaosrk. puTrhiteystoonclk)soalnudtiodni(tne 1f2o5-1m0L0LOmLP.E wbiatlhesm)ethan1o0lbeinstaor1ed00m.om
rdeaftreiogferparteopraraatti2onC,to 6C and is stablefor a maximum period of & months fram the.
lmPrLueproaferetwhaiethf1o0rmt0eitfghiac/antoLilonsstFtorcaaknmdsaotrlhduattoifosno1lut0toigoa/n,m1L0p0.r-em(pL1a.0r0ev0oalfnugom/ermtLrfi)iccoafftliPaosFnkOasAntdabnydbarairdnddgionofgp 011.00 4megt/hmaLno(l10i0n nvgol/uLmetbryicdfillatsikng 100 mL of the LO 4a/mL. solution 1a 100 mt. with
Psytrpaeenprdaacrraedrsb,a i2e5A0rdedndg1/wL0a0teexArt.raanocdfteptdrhoecceasl1is0b0trahtnriogou/ngmhLsttafhnodrteaixrftdircawacittitiohonnespaarclohuctesdoeuntre1o,0fi4ed0xetnrfmaosceteo1df samples
"
Npooitnet:foTrhwiasrsd,tandard is referred toas the "check standard" from this
raTnehaselpyozcnehsde.ecfkrTsohtmeatnchdheaerc2dk50sshtnoagun/ldLdaerbxdetrsiohncoctuleudddegcdiavleoifraaltreieaossntposontnseecnedwoiwrtidhtsihnineat1chh5%ssoeorff osfheeoxvterraacgtes
Page 13
POND The miracles of science
- 14
4. Method 4.1. Flow Diagram
The flow diagram of the methodi given below, followed by a detailed description of each
step.
MethodFlow Dicgram
(fortify samples designateMdeoassucarlei4br0amtiLonosftasnadmaprldesand fresh fortifications)
Cu Solid-phaTse Extraction Fil Volume (5 mL)
+
Lerms/ms
4.2. Sample Processing
saNlohloaswdeadmipstloeo cbpoermopaclleesotsweilenydg ttihosaewnq,eueuldineeardiadtefeodr. 0owfartoreoormomtsaemtmpeplmeeprsea.rtautrueHr.oew.eAvleSraa,mmppfllreeoszsesnmtuossraetmdpblereestfrhimegusessrtautgbeyed 5mlisxesdfiabnrde aasncyrovdiissicle0.s4o5liudfsilrteemrobveefdorbeybefiilntgersianmgplfehde fsoarmpelxetraschtriooingh a 25m cre, rietcaorldesdtupdriieosrsohoewxttrhaacttihoen meth is not effected by the pH of the sample. Sample pH is
Cafi5olnttweraroitlnegrytpyuep.seed wwfaaottrere,rquutlshierrdoauftgohirnatghheyapnpedrrecpwaaarrsbahtiginuoganrotdfhecsaarSmtPprEliedssceaaruntsdriincdgagleaisb,HrPamtLuiCosntpusmbtpea.ndparBroedpfsaorraeesduwsebely.l mrwaeattseh.hantohTleh,egnfuoalrsltdoawrcetadrtcbrayildl=gceti2wn5igtmhtLh~eo2ff5itlmytlpereHePdILCtwyagptreeardIeuswaiacntegteortnhieetlruiHalPet,eLChferponumm~pt2ha5etmc=trt2orfimHdLoPe/L.mCiar1ei1ow orfectoympemeLnudaetedr.thaAt ftlhoewcrartteriodfge2 fboe3wamsLh/emdi.na1s rdeesccormimbeenddsebdave, afte filtering = 2 iene 4.3. Somple Preparation a. EKcoanncothwrnoblcaot(ncmcheetnohtforsdaatbmilpoalnneskstuoesxivtnegrroifcfiytletpderro(ectdyepdtiiycrpaaelllyrew2ca0otoveerr)lyeasfnsd)ofmswuhsoetrbeianatcgclehundtecaotnlteraoslts ofnoertrfeeangenmtn
Page 14 --_
BORD The miracles of science:
1S
5. EFioeclhd sbalmanpklsedmousntotbneeeexdtrtoacbteedexitnrraecptleidcaitneruenplleicsastet.he sample is marked asa field blank
. Each sample extracted (except for field blanks) must also be separately
fortified at a known concentration and
verify recovery. Matrix spikes should
carried through
range from 500
the procedure to
ng/L to 500,000
ng/L.
level
If the sample residue is
at which the sample was
found to be significantly higher than the
initially fortified, the sample should be
fortified at a higher concentration (up to 500,000 ng/L) and re-
extracted.
4.4. Extraction
1 nMeeeadseudr,ere4p0lamcelIodfasnadmmpilxe wienlflo, 50 mL polypropylene centrifuge tubes (fortify as N0woe0ltdle2:to00dFe,o-rLac1otaipfvaawt2ae5tc0ehrmlogsr/ainmmep.,lesbsoeadfniourdmeanhfyoirstswiuafltifceaarttiesoasnmoplluteisosnuosptehcete4d0tmoLcosnatmapilnecahlnorimnei,
2. CondfiitlitoenretdhteyGpesTSPwEafcearrt(r2idgdersop(/1s9ec,)6,mLD)obnytpalstincgol1u0mnmirunmedtrhy.aral fllowed by 5 Ncootluem:n Ftoorrutnhdeefyololfowainnygtsitmepes, maintain a ~1drop/sec flow rare. Do not allow the
43.. LWaosdhswaimtphle~o5nncLo.n4di0t%iomneetdhaCnyolSPinEwcaatretrr.idgDei.scoDridscaalradtee.lute. 5. lute with 5 mL 100% metharal. Callect 5 mL of ehate info graduated 15 mL 6. pAonlaylpyrzoepyslaemnpelecsenutsriinfguegleetcutbreossp(rfaiynaLlCv/oMlSum/eMS=,5 mL). NVOoTLE=:40SaAMLPL>ESFIANREALCVOoLN.CE=NT5RAnLT.ED 8 A FACTOR OF EZGHT DURING THE EXTRACTION: INITIAL
45.1. LC/MS/MS System and Operating Conditions (Electrospray)
MInatsesrfSapceec:: ElMeicctrroomsapsrsaQyu(aMtitcrroomUaistsi)na (Wiramass)
Computer: HCaOrMvaPrAdQinPfruosfieosnsipounmapl W(oHrakrsvtaartdiIonnstArPu2me0n0ts). for tuning
Software: Windows NT. MassLynx 3.5
Hoc HewlettHPPacQkueartdP(uHmP)pSeries 1100
H1P VAuatcousuammpDleegrosser
Naotttea:cheAd
4on-xli1n0e
rm hyHpPerCcoalrubmndrOovpe-inn after the purge valve
gaunodrdbecfaorrtreidtghee
(sKaemypstloenei.njpeacrtotr#po8r4t40f1o7t-r4a0p0)aniys
residue contaminants that moy be in the mobil phase and/or HPLC system.
CHoPlLuCmnCoTleummp:eratGuree:neC3o5s(TCionses Chromatography)2,. mm x 50 rm, 4
Page 15 ee
POND The miracles of science" 16
MIonjbeiclteioPnhaVsoelu(mAe):: Mobile Phase 8).
15 4 2MemthManAamlmonium Acetate in Type I water
Time %A
%B Flow Rate
00
60
40
(mL/min)
03
04
60
40
03
10 10
90
03
70 10
90
75
0
100
03 03
20
0
100
04
95
60
40
135
60
40
04 04
140 60
40
03
150
60
40
03
dpTietfrffmeoarremynatnbceem.annuefcCaeocslstuaumrrnyesrstwoi(tKhaedyjdsuitsfotfneertehnBetetodHsiPimLleCnCsisgornaesdtic(e)engt.coiu2nl1dorxbdee3r0u)sfeaodn,dopprtaloismvoiizdceeodlieunqmsutnirvuafmlreeonnmtt
chromatography is obtained.
Lonsmonitored:
Andlyte Mode Transition ApReptrenotxiionmaTtimee
PFOA
Negative
Monitored 413-369
~5 min.
eTthc.e rDertiefnttiionnrteitmeenstimoanytviamreys. (ounpafo+da2y t1o%)daiys abcacsiesp,tadbelpeewnidtihnignonanthaenablyatticchalofrunm,oboisleopnhgasaes
bfehgeindnriinfgtacnodnteinndueosf tthheroaunaglhyttichael erunnt.ire analysis and the standards are included at the
4No5t2e:. AnElfxearmnpatlieveMLSC/OMpSe/rMaStisnygstPeamrmaamyebteeurssed once demanstrated fo be equivalent.
lTihkeelyfovlalroywifngroMmSionpsetrrautmienngtvfaoluiensstarruemepnrto.vidTedheasseavnaleuxeasmpmlaeyonallys.o cAhcatnuagle vaavleuresfwiimlel meovsetn
gwrheeanteasntalsyeznisnitgivsiatym.pleFsoronextahmeples,amheexicnpsotrlueme1ntv.oltaTgheemvaaylubees sarfe fcoha2ngVe:dhofoweovpetri.miazseHifmoer
gtoimees aons,etheixsaapnoalleyz1edvowilltlagbee dmoacyumbeentinecdrewaistehdeaorchddeactraeasseer.d. The exact values used each
coTofhnetPamFianOisAnsg(sa4pt0ec%1t0rmoyemlt/ehitmaeinnro,lisuastniudnnge6da0nf%oir2nftuhmseioaannmaplmuyotmnepi)buymviiaancfaeutsTiant*geaian~tfo00..a22 SgmtL/r/memaLimnsotffalnomdwoabrriadltees.olpuhtTaihsoeen
analyte is initially tune for the parent ion and then funed for the product ion. Once she
Page 16 ee
The miracles of science: "1
tihnesntruusmeedntduirsitnugnerdo,utitnheeanoapltyisniis.zed parameters are saved as a tune file, Ths un file is
Analyre
PFOA
Dwell (5)
02
Collision Energy (ev) cone(v)
10
10
Source
Capillary
Hexapole 1 Aperture 1
Hexapole 2 Source Block Temp. Desolvation Temp.
Analyzer
LM Res 1 HM Res 1
Energy 1
Entrance
Exit
LM Res 2
IHEMneRregsy
2
2
Multiplier
Gas Flows
Cone Gas Desolvation
set
3.0kV
ov ov
ov 100C 300C
set
14ov "ov
1v
2V 2v
14ov
12400vv
700 V
ser
~130 L/hr ~750 L/hr
Pressures 453.
Read back
GasCell
~3.0e-3 mbar
Calibration Curve Procedures
a. Ippnrrjeoepccaetrsest)eh,de iisnnatommeatthareliiLxquCo(/trMa(nSbg/eitMngwSee(fntrhoe1m0fittnhoael2c5loon4wce)esntotrfaleetviaeoclnhssetoxaftnrtdahacertsdeedfcoaclailtbihrbeartaithoiinognhsesssattnadnodleravdresdl will be equivalent to 8x the. concentration of the imitial standard due 0 the.
b. UicnsocenlcuedlnietndreaartiniosntthadenudraicnragdlcSucPluEa)rt.vioens offortqhueanctailtiabtrioant.ion Tchuervez.eroLcianleiabrrastitoanndsatradndcaurrdveiss nooret gcaelnierraattieodn fsotranedaacrhdacnoanlcyetentbryatliionneaursirneggrMeasssisoLnynuxsin3.g3 1/(xorweeqiugihvtailnegnto)fspoefawkaarreeasyvesrtseums Any calibration standard found to be a statistical outlier by using the Huge Errar Test,
Page 17
CWPOND The miracles of science
1%
ee
oumfamytbheebretooetfaxlcelxuutmdrebadcetrferdoofcmoexlttbhraeatcictoaenldcsistatataninoddnaarroddfsshtnhaeetccmraaloiybroaetieonxcciutrveed, muHsohwemvrero,xctehnedt5ot0a%l . TT2o0hk9ee8n5c)ot.roreaIldfajtucisaotlnbircnaostetifrofuinmcerienestnutolpt(esRr)afatlfiloona.uctaaslnibdcreattthihosnsetacmnuidratvressd,shgoeernnehraeaptpeerdloepmvruaiasrttteebsettepo2sf09es9et2e(op3e7
should be reanalyzed
a. coInntjercotl.tehte. snomtetahleiqLuCot/M(Sbe/tMwSeesnyst1e0m.to 25 1) of each standard, sample, recovery,
. SLSttOaaGnnddaalrerdvdesslacoorrrrpbereselpapowan)rdeimdnugbsyftosbpaeitkilinengcalftuidfeeidreeiodnrfamynopreeneclcwyotanitcceoerlnrasanetdtipo(rnnooceteeeslstishneg(tssttamhrteiincgoowittathcretehrse
Asansmepetlnetfsio)rlelosweetdobfyexetxrtaracctteeddcaclailbirbartaitoinonstsatnadnadradrsdsshionlfrdsbpeerisnefdecstpedparoxthiemtbeegienvniensg 5of
te1nh0teisreaenmpdsleeotfs o(af10seaaxmctpcrloaeucntsteetd.focraIlnibsireatctiohonend scstaesaten,dofaerxedtxsrtarmcaotcyetdebdecasnlatnerdaetriddo)n,astasAnhsdeoobnreassirmtaueisonsrgubemencd,hooonn
fTIihnrjesetctacnotdnhcelenoctshreiacntkjieoscnttaionondfairnedaacsnhacmepslwaeimtpsheltlefaocthifseitcaotfioenx/tcroancttesal is determined from the
sbsartmaapcnlkdeeastrdorceugsripvvoeen,saebrsaessoefpdocntsnheehwreietshpiiednaukethafroesuatnadonfdinsaraedcahccahurrsvleaymtrpealnegeTsehre. stIofndnoercdesrsearsyp.onsdeistheowtlhse
5f.. TEFoxhrtetriatfocittcaaeltdihsoonalmdrpielncgeosvfemirmuiesetsbebftelwisentegonrweisdtahmripenlfe7ri0cglfeloreac1tt3eid0o7nbeaatnewdecaeonnnls2yi:dsCesr15emdu6sactCcnuepntttaebsxlceeeieds14 days
h. wCFioherilrcdehssppaeomanpkdlsiensagrieanndawelhtyietcechtreeeidtteahnettrtiohnneoicpmoeraerkeswsipolornbpdeeiarnkgesapnloaelrsysttaheesadrneNtetOnhte(iroMontDsLemraeerceshtdaeetdtaecestealdmemeatiTetohne
the LOQ and
quantifiable).
greater
than
or
equal
fo
the
MOL.
wil
be.
reported
os
NG
(ror
C5To0hlel0eganen,ga/lPLyAsiYosfanpPdeFrbOfaAortmieneddweddlrulirnikwniagntgetrhw.aetetmrae.pt(hwfoadrtoedam,evoaenlldopmsgetrnroteceairmnycwlsaurdteee)dr,f(oerotliefcirceadtsionlsocaaltly.10S0taonrde
4T.h6e1..f_olACochwcirneogmpacttraiotngecrriaeammCmurusistttebsrehiomawf fpoaeknsoufreatdhaeugphrteesrenacne oaff P3F69OAam froma parent of 413 d(aim3su6c.9rimaTimhnuea)te41r3ebpeartewmseeenpntasrleitnntheeacrolraorsnesdspobofrnadncscahr1eb0do.nthfedoirPemxFisOdeA.ofanPiTFohnO.iAs:whsirslahenetrsehifieoorndeo.wuopugoldth.nieofrs present wuld be included m he calulrion
Page 18
QUADRD The miractes of science"
19
2. ccMooentttahaiionndessbPalFannOkAtshemaufnslttevrieoltssocgfornemtauatsietnrPbtFehOraAen-a5otx0tlrnegvoe/tlLes,deTahetnear ntehwanbtlhaenkLOsaGm.pleIfmaubrlabnek
3. sRKaenmcopowlvneersvailesusehso.oufldTcofbnet.rcoorlnet-srepoxilktesaspcitakenedd.amlastAroniuyxtssmipaditekretishxemasucspcteepbteoaubbtlseeitdmweiees7n,07-h01-e310e3%n0t%srheoofustethreboiefr
4. eATevnsaytl,ucmaotleybdrbabteyiotenhxesctlanauandldeyadsrtfdfrofoomduenttdheer1m0cialnbceeuliaaftrsieto.anetxiotsftriahccaetliaocanulliisebrrwaatbriyornavncituenedg.theHoHwusgeer,Erhroer
teoxtcaelednu2m0b%erofoftheexttrotaacltendumcbaelriborafteioxntrsatcatneddarsdtsantdahrtdscojuelcdtebde.exuded mast pon
5. osT2a0eh.me9pT8loc5eko)ser.rnse1l1ha0atciadaodinjlbubcesrotaertfeiifonainscntieyrezsnutmeaedt(nRet)fafapolrerocauaitlsoiibndraeottinohdnestcehuermvsietstasngdeeanrendrsatoaepdprhmouepsitraetbleeevs2at0ne.tp9ss9e2mracohf
6. rrTeehsseppoornnessseepoffnarslolems ftorhuaetms2id5te0hetnhgce/hsLee.cckamlsiitbsraa,ntdiaaornpdpsrtosaphnrodiuaaltrdedbcseotnewcpiestnhtimrnuast+tio1nb.5e%tIooffketnthheetsotaavsneudreapcgeee
airnnesdstpirofunmwseaenrtrisaonspttieelrdla,toiunotenswiadsnetdahctkeh,e+fsor1et5i%ofifctashtaeimnopnlseatsnadnsdehaxrotdurdapcrbetepeadrreacatanlilioybnzrsaetdsi.honouTsFdkttbiheoenrssetvsahenemdvenidsd
7.
oReetpernetpiaorned.times between standards and. 2 % within on andytcal rn. If retention
siommeploesftmuesxtceerdts
thdirsifmtinmewriethitnhoann
ealyicl run then he se must be reralysed
SmIponescostrsrdpeoermce1tuoesrivneugrsiiafnnygimanafsuNssailor/neRspboul/umCtpsiosnt,eunai1n0cgaalsobflrltaoitwoinoa,ntechToerfcek1T0iusnsiipnmegirnfs.oormoAennd MwieSsemkcluoysmeondratohmiee8m0enses tM1hS3e8Z0:foal8mlou4w9iin,sg1pe13r2fp,oera1mk7es2d8oa,ren3bf2oo2tu7hn,dM4wS7iL2th6ai,nnd6+2M20S..252,.a7m7T2h.oe4f,cha9l2ei2br5r,atkin1oo0n7w25n.2mc.oosn1ss2ei2ds2e.1ornendbdaocrc13e7p5M.tS0aTblean1e5 aannyotohfertchaelimbroastsieosn acrheecokutpseirdfeofrhmeed+ 02 amu, then he instrument il oe recaioened ang
c4To.h8me.mefonlcPleoewmriefnngotrtomwfoaoncacrlieytseiCrsiraiwtmheuresitausbineg his method
paenrifnosrtmreudm
eonstatoiosnyssote
mu
p
suhitaatbi
lhiatsy
ntaetst,cebnefuoered
Tfhoe
R(F5ui0nrsngCa/rLi)tseitrnaimonndierxd asnodloutbiaoninaansigLnaCl/tMoSo/iMeS ractoiroroefsptohnedi4n1g3 --to369thseraneisttiomnaotfeda lLoOoQk
5in1strcuommepnatroepderfaotiangrpeaagreanmtetbelrask(o Iinctrehaisecrsihteerjieocntcioannvtolubmee.meft.spapprtampirsineen) d chon
Page 19
APOND The miracies of science
K0
TRountahesehtigohfessttacnodnacrednstroaftifoinveleovrelmofroebceonicnecnlturdaetdioinn tlehveelsa,nalfyrsiasm. ofGoernebrealtoew athsealLiOoGr,aiuopn ilcenuatrsevtres0fp.oe9rr8sP5eFdfOorA PaFnOdAo.btOanicane ltihnesarcrrietgerreisosnioin mweitt,h saacmopelfefsicmiaenytboef adneatleyrzmeidnaittiohn s(tRa)rodfraast
dB4ee.tf9eo.rrmeinMasetaimtophnleosshdocuDalendtbebeectpiaenroafnloyrLzmieedmd.itoDnTehtoenerpirmnoisctneraduutmreienotf,nolalowMeedthcoond bDeetfeocutnidonin L4i0miCtFR(MPOoLr)t
136 ona
qAupapretnedrilxy
bB.asiTshe
MDL
should
then
be
performed
on
each
instrument
used
for
anelyss
p44r.os1ce0et.duorfTei2im4neaspRapmerpqolxueisimra(et6edlsytfao2nr-d3aArnhdoasulraysnsdbiys18onseamppelresosn).caTnhebe.LtCo/kMenS/tMhSrouangahlytshies (esxrtarnadcotridoen and 18 samples) will ake opproxinately 7 hours.
5. Calculations
a. UtsheeEqsutaatnidoanrd1 1c0uCravlecul(aitneetahre raemgareusnstioonf. PpFaOrAamfeotuenrds)(ingnegn/eLr.abteadsedbyan tpheeakMaarss) uLsiyng software program. Equation 1
PFOA found (ng/L) =(Peakarea-oipnteercept) x DF
DF = factor by which the final volume was diluted, ifnecessary.
b. Fcoarlcsualmapteletshefopretirfcieendtwrietchovkenryo.wn amounts of PFOA prier- to extraction, use Equation 2 fo
`Equatio2n:
Recovery (1%) =(total analyte found - analyte foundi controlY/anlyte added Jx 100
T Lota anaT lyfound voa1y)eadsenadyotewohu)nd in control gr] Note: Subtroct analyte found in contro (g/L) from analyte found (g/L), if applicable.
b. wUesieghEtquraattiioonof3AtPoFOca/lPcuFlOaAte the amount of APFO found (in ng/L) using the maleculor
Page 20
WROD The miracles ofscience:
Equatio3n:
#70 found(r9/L) = PFOA FoMu MWWAAn PPFFOO((d 33D1))
ha
i)
MW PFOA (414)
Niontneg:/mcLalicnusltaetiaodnosfcogu/lLd swhoenbepeprefrofromirnmgedhein panoaleoirs.ng/mL. if concentrations were assigned
daFreoetrercdetepetoder)cttiendgSapamutprlpteshseesin,cowfrhirieecsdhpospnadmeipcnlkgesaarinnaelwydhteietcehrcetetieetdhnetrai.fonntohfpmeeCaokrswrieolsrpbpoeendaikrnsegpolearsntseldythtaoesnrtNehGfe,eMi(DorLnr time that are less than the LOQ and greater thano equal 1o the MOL ill ow reparted a
NQ (not quantifiable).
.
Safety
The analyst should read the material safety data sheets for all
psrteacnaduatridosnsanwdherneahgaenndtlsinbgesftoarnedpaerrdfsoarnmdinrgeatgheinstmse,thiondcl.udiUnsg ewournkiivnegrsianl fume hoods and wearing laboratory coats, safety glasses, and gloves.
Z REFERENCES
hE7l2lei6s,.A7nD3a.A1l.y;siMsarotfinF,luJo.rWi.n;teMudiArc,iDd.sCi.n EnMvaibruornym,enSt.aAl. WDaetveerloapmmpelnets,ofAanl'FhNeMmR 2M0e0t0h,o5d2for MPeorofdlyu.orAin-ateKduaSnu,rfWaLcCt:antMasrtininS,urTfWac:e MWaaitrerD.SaCm.plMeasubryyT,wS.A.ndeDpeetenrdmeinntatAinolnyroifcal 2001. 73, 220.2206 Techniques: Liquid Chromatography/Tandem Mass Spectrometry and "F NMR. Anal. Chem.
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QUPOND The miracles of science
a