Document e76NMpd2rEKy7xoqBXOKD9m3E
A R226 oo2y
Attachments to April 21, 2000 letter to C. Auer from W. Weppner (Tissue Extraction and
Analysis Methods)
1
3M Environmental Laboratory Method. Extraction ofPotassium
`PUesrifnlguoHrPoLoCc-tEanleescutlrfoosnparteayo/rMOatshseSrpFelcutorroomecthreym.icaMletChoomdpoNuunmbdesrf:roEmTSS-e8-r4u.m1.forAAdnoapltyisoisn
Date: 03/01/99.
2.
3M Environmental Laboratory Method. Analysis of Potassium Perfluorooctanesulfonate
or Other Fluorochemicals in Serum Extracts Using'HPLC-Electrospray/Mass
Spectrometry. Method Number: ETS-8-5.1. Adoption Date: 03/01/99.
3. 3M Environmental Laboratory Method. ExtractionofPotassium `Perfluorooctanesulfonate or Other Fluorochemical Compounds from Liver for Analysis Using HPLC-Electrospray/Mass Spectrometry. Method Number: ETS-8-6.0 Adoption
Date: 07/22/99.
4.
3M Environmental Laboratory Method. Analysisof PotassiumPerfluorooctanesulfonate
o`rSpOetcthreormeFtlruyo.roMcheetmhiocdalNsuimnbLeirv:erETESx-t8r-ac7t.s0.UsiAndgo'pHtPiLoCn-EDlaetcet:r0o7s/p22r/a9y9/.Mass
s. 3M Environmental Laboratory Method. Extraction ofPotassium
Perfluorooctanesulfonate or Other Fluorochemical Compounds from Urine for Analysis Using HPLC-Electrospray/Mass Spectrometry. `Method Number: ETS-8-96.0. Adoption Date: 07/28/99.
6.
3M Environmental Laboratory Method. Analysis of PotassiumPerfluorooctanesulfonate
or Other Fluorochemicals in Urine Extracts Using HPLC-Electrospray/Mass
Spectrometry. Method Number: ETS-8-97.0. Adoption Date: 07/28/99.
000997
3M E NVIRONMENTAL LABORATORY
METHOD
EXTRACTION OF POTASSIUM PERFLUOROOCTANESULFONATE OR OTHER FLUOROCHEMICAL COMPOUNDS FROM SERUM FOR ANALYSIS USING HPLC-
ELECTROSPRAY/MASS SPECTROMETRY
Method Number: ETS-8-4.1
`Adoption Date: 03/01/99
Author: Lisa Clemen, Glenn Langenburg
Approved By:
AI
Laboratory Manager
V;ideo fo
Group Leader
Revision Date: 4/2 7199
"for tss
Date
4/20/79
Date
a A oss `Technical Reviewer
4/26; 6 Date
1.0 SCOPE AND APPLICATION
11 Scope: This method is for the extractionofpotassium perfluorooctanesulfonate (PFOS) or other fluorochemical compounds from serum.
12 Applicable compounds: Fluorochemical surfactants or other fluorinated compounds.
13 he valdsion reper Matrices: Rabbit, rat, bovine, monkey, and human serum or other fluids as designated in
wortass
Eacieprospanarrm Sem
00095o8g Lot10
2.0 SUMMARY OF METHOD
2.1 T(hPiFsOSm)etohroodthdeerscfrliuboersotchheempircoacledsuurrfeacftoarnetxstrfarcotminsgeproutma,sosriuomthpeerrfflluuiodrs,ooucstianngcsaunlfioonnate
pfaliuroirnogchreemaigceanltsawnedrmeetehxytlr-atceterdt:-bPuFtyOlS,ethPeFrO(SVAI,BEP)F.OISnAtAhi,s EmettFhOoSdE,-sOeHv,enPFOSEA,
M56, and the sample
surrogate standard and the analyte ion
(see pair
3.0 Definitions). is partitioned into
An ion MBE.
pairing reagent is added The MIBE extract is
to
rmeLmoovfemdetahnadnoplu,t othnetno failntietrreodgtehnreovuagphoraa3tocrcupnltaisltidcrys.yrEiancgehaetxttarcachtedistoreaco0n.s2tuitmutendylionn1.0
filter into glass autovials.
22
These sample methods.
extracts
are
analyzed
following
method
ETS-8-5.1
or
other
appropriate
3.0 DerivrTIONS 31 PFOS: perfluorooctanesulfonate (anionofpotassium salt) CF,SO, 32 PFOSA: perfluorooctane sulfonylamide C,F,,SO;NH,
33 PFOSAA: perfluorooctane sulfonylamido (ethylacetate C,F,,SO,N(CH,CH;)CH,CO, 34 E{FOSE-OH: 2(N-ethylperfluorooctane sulfonamido)-ethyl alcohol
CJF,,SO,N(CH,CH,)CH,CH,0H
3.5 PFOSEA: perfluorooctane sulfonyl ethylamide C,F,,SO;N(CH,CH,)H 3.6 M556: CF,SON(H)(CH,COOH) 37 Surrogate standard: 1H-1H-2H-2H perfluorooctane sulfonic acid
4.0 WARNINGS AND CAUTIONS 41 Health and safety warnings
4.1.1 Uhasnedluinnigvearnsialmaplretciasusutei,onwsh,iecshpemcaiayllcyonltaabionraptaotrhyocgoeantss., goggles, and gloves when
5.0 INTERFERENCES 5.1 There are no interferences known at this time.
6.0 EQUIPMENT. 61 Tachceepftoalblloew.ing equipment is used while performing this method. Equivalent equipment is
6.1.1 Vortex mixer, VWR, Vortex Genie 2 6.12 Centrifuge, Mistral 1000 or IEC 6.13 Shaker, Eberbach or VWR
ExtracoEftTiPSFo8On4Sf1romSerum 000999 Page 20614
6.1.4 Nitrogen evaporator, Organomation 6.1.5 Balance (+0.100)
7.0 SUPPLIES AND MATERIALS
71 Gloves 72 Eppendorf or disposable pipettes 7.3 Nalgene bottles, capableof holding 250 mL and 1 L 7.4 Volumetric flasks, glass, type A 75 1.CHEM vials, glass, 40 mL glass 76 Centrifuge tubes, polypropylene, 15 mL 7.7 Labels 7.8 Oxford Dispenser --3.0to 10.0mL 79 Syringes, capable of measuring 5 uL to 50 pL 7.10 Graduated pipettes 7.41 Syringes, disposable plastic, 3 cc 7.42 Syringe filters, nylon, 0.2 pum, 25 mm 743 Timer 7.14 Crimp cap autovials and caps 7.15 Crimpers Note: Prior to using glasswareandbottles, rinse 3 times with methanol and 3 times with
Milli-QTM water. Rinse syringes a minimum of9 times with methanol, 3 rinses from 3 separate vials.
8.0 REAGENTS AND STANDARDS 81 bTeyMpielli[-rQeTMagenwtagtreardeanwdatmear,yMbielplr-ovQiTMdeodrbeqyuiavMailelnlti,-QalTl OwaCtePrluusseTMdsiynsttheismmethod should 82 Sodium hydroxide (NaOH), J.T Baker or equivalent 83 Tetrabutylammonium hydrogen sulfate(TBA), Kodak or equivalent 84 Sodium carbonate (N&,CO;), J.T. Baker or equivalent 85 Sodium bicarbonate (NaHCO), J.T. Baker or equivalent 86 Methyl-T-Butyl Ether, Omnisolv, glass distilled or HPLC grade 87 Methanol, Omnisolv, glass distilled or HPLC grade 88 Serum or blood, frozen from supplier 89 Fluorochemical standards
89.1 PFOS (3M Specialty Chemical Division), molecular weight = 538 8.9.2 PFOSA (3M Specialty Chemical Division), molecular weight = 499
ExtractioEnoTfSP8F4O.S1from Serum
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001000
893 PFOSAA (3M Specialty Chemical Division), molecular weight = 585 89.4 E(FOSE-OH (3M Specialty Chemical Division), molecular weight = 570 89.5 PFOSEA (3M Specialty Chemical Division), molecular weight = 527 8.9.6 M556 (3M Specialty Chemical Division), molecular weight = 557 8.9.7 Surrogate standard: 4-H, perfluorooctane sulfonic acid (1-H, 1-H, 2-H, 2-H
CF,,SO,H) molecular weight = 428 89.8 Other fluorochemicals, as appropriate 810 Reagent preparation NOTE: When preparing larger volumes than listed in reagent, standard, or surrogate
preparation, adjust accordingly. 8.10.1 10 N sodium hydroxide (NaOH): Weigh approximately 200 g NaOH. Pour into a
1000 mL beaker containing 500 mL Milli-QTM water, mix until all solids are dissolved. Store ina | L Nalgene bottle. 810.2 1 N sodium hydroxide (NaOH): Dilute 10 N NaOH 1:10. Measure 10 mL of 10 N NaOH solution into a 100 mL volumetric flask anddilute to volume using
Milli-QTM water. Store in a 125 mL Nalgene bottle.
8.103 0o.f5TMBAteitnrtoabaut1yLlavmomlounmieturmihcycdornotgaeinnisnuglf5a0t0e (mTLBAM)i:llWi-eQiTMghwaaptperr.oxAidmjautsetlyto1p6H9 g 10 using approximately 44 to 54 mL of 10 N NaOH (While adding the lastmL of NaOH, add slowly because the pH changes abruptly). Dilute to volume with Milli-QTM water. Store ina 1 L Nalgene bottle. 8.10.3.1 TBA requires a check prior to each use to ensure pH = 10. Adjust as needed using | N'NaOH solution.
8.10.4 0.25 M sodium carbonate/sodium bicarbonate buffer (Na,CO/NaHCO,): Weigh approximately 26.5 g of sodium carbonate (N2,CO) and 21.0 gofsodium bicarbonate (NGHCO,) into a 1 L volumetric flask and bring to volume with MilliQTM water. Store ina 1 L Nalgene bottle.
811 Standards preparation 8.11.1 Prepare PFOS standardsforthestandard curve. 8.11.2 Prepare other fluorochemical standards, as appropriate. Multicomponent fluorochemical standards are acceptable (for example, one working standard solution containing 1.00 ppm PFOS, 1.02 ppm PFOSA, 0.987 ppm PFOSAA, and 1.10 ppm EtFOSE-OH.) 8.11.3 Weigh approximately 100 mg ofPFOS into a 100 mL volumetric flask and record the actual weight. 8.11.4 Bring to volume with methanolfor a stock standard of approximately 1000 ppm (ug/ml). 8.115 Dilute the stock solution with methanol foar working standard 1 solution of approximately 50 ppm. 8.11.6 Dilute working standard 1 with methanol foar working standard 2 solution of approx. 5.0 ppm.
ExtractioEnoTfSPFSO4S1from Serum
Pagedo1f4
001001
8.11.7 Dilute working standard 1 with methanol for a working standard 3 solution of approx. 0.50 ppm.
812 Surrogate stock standard preparation 8.12.1 CW,eFi,g,hSOa,ppHroinxtiomaat5e0lym5L0-v6o0lummgetorifcsufrlraoskgaatnedstraencodradrdth1e-Ha,c1t-uaHl,w2e-iHg,ht2.-H, 8.12.2 Bring to volume with methanol for a surrogate stockofapproximately 1000-1200 ppm. 8.12.3 sPtroecpkarteo a s1u0rrmogLatveolwuomrektirnigc sftlaanskdaarndd. bTrrianngsftoervaoplpurmoexiwmiatthemleyt1hamnLoloffosruarrogate working standard of 100 ppm. Record the actual volume transferred.
9.0 SAMPLE HANDLING 9.1 All samples are received frozen and must be kept frozen until the extraction is performed. 92 Allow samples to thaw to room temperature prior to extraction.
10.0 QuaLITY ConTROL 10.1 Solvent Blanks, Method blanks and matrix blanks
10.1.1 An aliquot of 1.0mLmethanol is used as a solvent blank. 10.1.2 Extract two 1.0 mL aliquots of Milli-QTM water following this procedure and use
as method blanks, 10.1.3 mExattrraicxtbtlwanoks1..0SmeLe 1a1l.i1q.u4o.tsofthe serum following this procedure and use as 102 Matrix spikes 10.2.1 Prepare and analyze matrix spike and matrix spike duplicate samples to determine
the accuracyofthe extraction. 10.2.2 Prepare each spike using a sample chosen by the analyst, usually the control
`matrix received with each sample set. 10.2.3 `EAxdpdeicttioendalcosnpcieknetsramtaiyonbsewiilnlclfualdledinatnhdemmiady-rfaalnlgienotfhtehleowin-irtaianlgceoalfibtrhaetiionnitciuarlve.
calibration curve, 10.2.4 Pmrienpiarmeuomnoef 2matmraitxrisxpiskpeikaensdpmeartbraitxchs.pike duplicate per 40 samples, with a 103 Continuing calibration checks 103.1 Prepare continuing calibration check samples to ensure the accuracy of the initial
calibration curve. 10.3.2 ePrxeapmaprlee,,iatfaa msianmipmleums,eto=ne34c,onftoiurnucihnegcckhseacrkepperrepgarroeudpaonfd1e0xtsraamcptleeds.. For 10.3.3 Prepare each continuing calibration check from the same matrix used to prepare
the initial curve,
Extraction oEfTPSF8O4S.1from Serum
Page Sof 14
001002
10.3.4 The expected concentrations will fall within the mid-rangeofthe initial calibration curve. Additional spikes may be included that fall in the low-range of the initial calibration curve. This is necessaryifthe analyst must quantitate using only the low end of the calibration curve (for example, 5 ppb ~ 100 ppb, rather than 5 ppb -- 1000 ppb).
11.0 CALIBRATION AND STANDARDIZATION 111 Prepare matrix calibration standards
11.1.1 Transfer | mLof serum to a 15 mL centrifuge tube. 11.1.2 Ifmost sample volumes are less than 1.0 mL, extract standards with matrix
volumes equal to the sample volumes. Do not extract less than 0.50 mL of `matrix. Record each sample volume on the extraction sheet. 11.1.3 While preparing atotaloftwenty aliquots in 15 mL centrifuge tubes, mix or shake between aliquots. 11.1.4 Two 1mLaliquots, or other appropriate volume, serveas matrix blanks. Typically use the standard concentrations and spiking amounts listed in Table 1, af the endofthis section, to spike, in duplicate, two standard curves, for a total of eighteen standards, two matrix blanks, and two method blanks. 11.15RerafnegrestoanvdaltihdeatLiionnearerpCoarltiEbTraSt-i8o-n4R.a0n&geE(TLS-C8R-)5.fo0r-Vca-l1i,brwahtiiconhcluirsvtesst.he working 11.1.6 UstsaendAatrtdsa.chSmeeentSeDctaisoann13ai.d0 tion ccaallccuullaattienagcttuhaelccoonncceennttrraattiioonnssoofftPheFOwoSrkiinng. calibration standards. 112 To each standard, blank, or continuing check, add appropriate amountofsurrogate working standard for the concentration to fall within the calibration curve range 5 ppb = 1000 ppb. 113 Extract spiked matrix standards following 12.6-12.16ofthis method. Use these standards to establish each initial curve on the mass spectrometer.
ExtractioEnoTfSPF8O4S1from Serum
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001003
Table 1
Approximate spiking amounts for standards and spikes
Using 1.0 mL of matrix
Working standard
nL
"Approx. final conc. of
(approx. conc.)
analyte in matrix
TT Bank |
0.500 ppm |10[ 0005ppm
05..50000ppppmm
00..002150 ppppmm
55.0000 ppppmm. [10| 50.010500ppomm
S00ppm [5| 0250pm
5S00.00ppppmm. [1 [5 1| 0 0057000pmpmm
Sooppm | 20| 1.00pm |
12.0 PROCED!
12.1 Obtain frozen samples and allow to thaw at room temperature orin a lukewarm
waterbath.
122 Vortex mix for 15 seconds, then transfer 1.0 mL or other appropriate volume to a 15 mL. polypropylene centrifuge tube.
12.3 Return unused samples to freezer after extraction amounts have been removed.
124 Record the initial volume on the extraction worksheet.
125 Label the tube with the study number, sample ID, date and analyst initials. See attached `worksheet for documenting the remaining steps.
12.6 Spike all samples, including blanks and standards, ready for extraction with surrogate standard as described in 11.2.
12.7
Spike each matrix with
1 in that section, for the
the appropriate amountofstandard as described in 11.1,
calibration curve standards. Also prepare matrix spikes
or Table
and
continuing calibration standards.
12.8 Vortex mix the standard curve samples, matrix spike samples, and continuing calibration
samples for 15 seconds.
12.9 Check to ensure the 0.5 M TBA reagent is at pH 10. Ifnot, adjust accordingly.
12.10 To each sample, add 1 mL 0.5 M TBA and 2 mL of 0.25M sodium carbonate/sodium bicarbonate buffer.
12.11 Using an Oxford Dispenser, add 5 mL methyl-fert-butyl ether. 12.12 Cap each sample and put on the shaker at a settingof 300 rpm, for 20 minutes.
12.13 Centrifuge for 20 to 25 minutes at a setting of 3500 rpm, or until layersare well
separated.
Extraction oEfTPSR8O4S1from Serum
Page Tof1s 001004
12.14 Labela fresh 1S mL centrifuge tube with the same information as in 12.5. 12.15 Remove 4.0 mL of the organic layer to this clean 15 mL centrifuge tube. 12.16 Put cach sample on the analytical nitrogen evaporator until dry, approximately 1 to 2
hours. 12.17 Add 1.0 mL of methanol to each centrifuge tube using a graduated pipette. 1218 Vortex mix for 30 seconds. 12.19 sAytrtiancghe.a 0F.i2ltuerminntyolaon1.m5emshLfgillatesrstaoutao3vicacl soyrrlinogwe-vaonldutmreanasufteorvtihael swahmepnlenetcoetshsiasry. 12.20 `Lmaabterilx,thfeinaaultosovlivaelnwt,itehxttrhaectsitoundydantue,mbaenrd,aannaliymsatl(sn)upmebrefroramnidnggetnhdeere,xtsraacmtpilone.timepoint, 12.21 Cap and store extractsat room temperature or at approximately 4 C until analysis. 12.22 Complete the extraction worksheet, attached to this document, and tape in the study
notebook or include in study binder, as appropriate.
13.0 DATA ANALYSIS AND CALCULATIONS 13.1 Calculations
13.1.1 Calculate actual concentrations of PFOS, or other applicable fluorochemical, in calibration standards using the following equation: TLmLofosftsatnadnadradr+d xmcLoonfcesnutrrraotgiaotneosftsatnadnadradrd+ (inuigti/almLm)atrix volume (mL) =
Final Concentration (ng/mL) of PFOS in matrix
14.0 METHOD PERFORMANCE. 141 TfohrespmeectihfiocdMdeDtLectainodn lliimmiitto(fqMuDaLnt)itisatainaolnyt(eLOanQd)mvaatlruiexs s(pseeceifAict.tacRehfmeernttosMBDaLndreCp)ort 142 The following quality control samples are extracted with cach batch of samples to
evaluate the quality of the extraction and analysis. 14.2.1 Method blanks and matrix blanks. 14.2.2 Matrix spike and matrix spike duplicate samples to determine accuracy and
precisionofthe extraction. 14.2.3 Cinointtiailncuailnigbrcaatliiobnractuirovne.check samples to determine the continued accuracyof the 143 Refer to section 14 ofETS-8-5.1 for method performance criteria.
15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 hSiagmhplBeTwUasctoentiasindeirssp,osaendd iunsebdioghlaazsasrdpicpoentttaeiwnaesrst,e fisladmimspaobsleedsionlvbernotkweansgtleasiss cdoinstpaoisneedrsin
located in the laboratory. ExtarcactioEnoTfSP0F8Os4S1from Serum 001005 Page8of 14
16.0 Recorps 16.1 Complete the extraction worksheet attached to this method, and tape in the study
notebook or include in the 3-ring study binder, as appropriate
17.0 Artacuments
17.1 Attachment A, Extraction worksheet
0000000000000
172 Attachment B, MDL/LOQ values and summary
17.3 Attachment C, Calibration standard concentration worksheet
18.0 REFERENCES 18.1 The validation report associated with this method is ETS-8-4.0 & 5.0-V-1. 182 FACT-M-3.1, "Analysisof Serum or Other Fluid Extracts for Fluorochemicals using
HPLC-Electrospray Mass Spectrometry"
19.0 AFFECTED DOCUMENTS
19.1 ETS-8:5.1, "Analysisof Serum or Other Fluid Extracts for Fluorochemicals using 'HPLC-Electrospray Mass Spectrometry"
20.0 Revisions Revision NumbI er Section 12.21 Changed toReinacsloundeFsoarmpRleevisstioroangeat room temperature.
Section 12.13 Added theshakerspeed. Section 12.17 Final volume is 1.0 mL; not adjusted for initial volumes less than 1.0 mL.
Revision 04D/a02t/e99
ExtractionEofTSPF8O4S1from Sem 001.006 Page9or ie
Extraction Worksheet ETS-8-4.1
SMtaurdyi,
aSpiprrooxg.ue Spdpm|| apFpOrN0o5Exp.m| prFOoN5xpp.m | apprFo.C5M0 ppm| Comments
WBioxD#a. y
scual ppm|"acnal ppm| scwal ppm | #acual ppm
DatcSpiked/Anslyst
-- e wsv r po o r r ]
r wo [r |rp r ]
rrr
r rr r Hr S r r Sr Y 1|
r rr r rr rr
rT
r rr r rr r1
r rr rr r
r rr rr r
r rr rr r
r rr r rr r T
r rr r rr r r
r rr rr r
-- r rr r r r r
rrrr
eee ee
Ce
el
omsiee
[PeteTmLoroSsNezcOyOBMNCO bate
--ser saw
|
(DipemesnLotmeyiibueher
moa
vow
ET
A |
"Cont Cal Verifications wed same mat a for sd curve.
Atachment A
ExtroEfTaPSF8cO.S4t1foimSoennn. 001.007 Page 100114
MDL/LOQ values for rabbit serum
Compound
MDL
(ppb)
|
|
LOQ|
(ppb) |
Linear Calibration Range (LCR)
Approximate concentrations to be
used
for
preparing
the
[Pros [T7555[sSotpanbd-artdoCoaolipbryation Curve
|
[EFOSEOR| 114 |362 [spbtoooppo
|
Ta ee wi Soin. ont and a sr, Srey IGT vere wl Gaol determined. Two curves in each ofthese matrices were extracted and analyzed with the rabbit serum
curves to determine equivalence. Responses in the rat, bovine, monkey, and human were equivalent to the rabbit responses, therefore,their MDL and LOQ will be the same values as determined in rabbit
Please see LOQ Summary and MDL study in ETS-8-4.0 & 5.0-V-1 for further information.
Aachen: NDLILOQ Samy TUN.E.ss.a DA
Page thot1s
001003
a | ETSI
(ppb) (ng/mL)
(ppb)
Fores| omon | wsom |w|m sons|
ra | wean |ww| sem
am | mam
www
[oreor][oPoPn [|owTRnE |-- | wows| Compound: PFOSA
(ppb)(ng/mL)
(ppb)
Full Range
(ng/mL)
0993-976 | 493-976 [wo|sews|
[[ ie| rm e[osmo n|| mwmnn [|wors|| `Compound: PFOSAA
| | Em Rabbit Serum| ofstandards curve
Range
Range
ite
| worn[wom (ppb) (ng/mL)
(ppb) (ng/mL)
|aw
|oe|
ee |gn | mi | en| |
mts eTsehs nn 002003--
`Compound: EtFOSE-OH
Prepared range LCR from % Recovery
RSD
Fans
(ppb) (ng/mL)
was (ppb)
(ng/mL)
ams
[ieee| oe |ws| ow|e
Prepared range|LCRfrom | %Recovery
RabbitSerum ofstandards
curve
Range
(ppb) (ng/mL)
(ppb) (ng/mL)
RSD
Range
woe | ores
[oT
Compound: M56
Prepared range LCR from % Recovery
RSD
RabbitSerum of standards
curve
Range
Range
[Rime
[oon [ovo (ppb) (ng/mL)
(ppb) (ng/mL)
|
or |ms|
[wreo|on me[[oowm||owmon[|amweosn]|
001010
Ion Pair Standard Curves -- Fluids
SAnaParmelppyltdeaetm)e(a:st)r:ix: TMaertgheotdarneavliystoen(9::
EFqiSuntaialnpsdmaerondltnuvnmaubemnebdrne:TrN:t: Blank Muididencfer:
FC mix std approx. 0.500 ppm:
FSFCuCrmrmioigxxatsstetddsaatdppaprpopxpr.ox55r..00.00o1p0p0pp.mp::m:
Actual conceonfsttanrdaardstinithoe FCnmsix
SdomLmc Ssicmobm sSugconme Sieugml m Sgiemone vSgonom||spAmht edmFiaiLlvol
[[00350000|| 00550077 ||003.5232 | |0os0o1r ||05sa2l1 || oossoorr || o0o01z00[|LO10I2S5 ||
[[530000| 50so77 || 533322 | | ssoorr||ss2a1i|| ssooir_|| o00o0r5 | TtoOsO||
[Seo | soy [500[Sox
| |
532 ssz
| |
sor|sai | soi|sal|
sor s01
|0020 [TO | |0005 [TOW |
[soo soa |_s2 | sor| sar| sor | ooo| ios|
[S00 | soi| [Soo[son|
332 532
| sod| sai | | soa | sat |
sor| sor |
00s TO0X | 00x | Tow]
oe CalculatedrcooncaentraPtRiOonSsAoKf staEnWdOaSrEd~s iPnFtOhSeEsAamplNe mSatrixSaag AT
om : .
re Sam el
[4 I 9 [50J 0 | s2 | 49 1soiTsp[10| | o05|
[[i2oas8 | saosoi| |2e3| | e a8| |aS020 [|BslFa|| Falcone
[Co 376e [98m 9 | Toioe |a97e8 [ |m99r3[[Tooox | |
ngmi 500
[aEn E [so | sa 0J
| es | sJ o [sn |
7
C0
Validated ranges- approximate concentrations
[[RaSbebim | 5P0F0O-1S0_0___5P0F0O-1S0A0_0 |_P0F6O-S10A0A0_|_E5W0O0-S1E00O0M|_F50F0O-S10E0A0 | 50W0i-s1s000||
=|
[Miokey&Plasma| Estimatesonly.Usevaluesformbon.
|
tachment C: on Pai Sandard Curves ExtractionoEfPrEsGSaanfom Serum ofrros
Page dre 001011