high lc qtof ms for the of unknown contaminants · pdf file28.09.2016 · high...
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HighResolutionLCQTOFMSfortheScreeningofUnknown
ContaminantsinBiologicalandEnvironmentalSamples
M.Wang,E.Parry,E.Houtz,S.Salihovic,S.Anand,Petreas,M.,ParkJS.
BiomonitoringBranch,ECLDepartmentofToxicSubstanceControl
9/28/16
WhyQTOFScreening?Beyondthetargetedanalytes list
Agilent6550LCQTOF
The Agilent 6550 iFunnel Q-TOFsupports the Agilent Jet Stream ESI and the Dual Agilent Jet Stream ESI.
Massaccuracy:subppmlevel
Highresolution
Nolossofdataacrossentiremonitoredm/zrange(1001700,smallmolecule)
PresentationOutline
SystemandProjectsOverview TargetSuspectScreening>NontargetSuspectScreening> SemiUnknownapproach
ProjectsSummary(biologicalandenvironmentalsamples)
1.SFOwastewaterscreening2.Catserumscreening3.FoamProject
FBF(findbyformula)algorithm
MFE(molecularfeatureextraction)algorithm
1.TargetSuspectScreening:SampleswithinterestedTargetedChemicalFamily
2.NonTargetSuspectScreening:notlimitedtooneparticularlyfamilybutexpandtobroader
nontargetcontaminants
3.SemiUnknownApproach:beyondcurrentdatabases
ProjectsOverview SFOwastewater:historicalusageandcurrentannualtesting
ofAFFF(aqueousfirefightingfoam)atSFOresultinhighconcentrationofPFASsinwastewater.Whatarethoseprecursors/intermediatesintheWWinthetreatmentplantsbeforedischargetobay?TargetSuspectScreening
Catserumexposome screening:apilotprojecttosetupbiomonitoring/serumsamplemethodontheQTOF.Convenientsamplesof5HTcatsand5Normalcatsserum.compoundsprofilescomparisonbetweenHTandNormalcats.NonTargetSuspect+SemiUnknown
Participateinthesaferconsumerproductprojects(SB1019labeling):helpconfirmifthereisnopresenceofOPFRs,aswellasexpandidentificationofotherFRspresentinfabric/foams:intheprocessofsummarizeintoastandardmethod.TargetSuspectScreening
Library1. organophosphateflameretardants
(OPFR)library:922. ExpandedPFASlibrary:2443. InHouseLCMSlibrary(using
consumerproductchemicalsindex,combinewithourinhousechemicalsanduptodateliteratures):~2500
4. EPATox21library(DSSTox_ToxCastRelease_20151019):~9000
5. EOAlibrary(Environmentalorganicacids,CollaborationProjectwithUCSF):744 3.GoldSmith et.al.FoodChem.Toxicol.,2014,269279
4.Richardet.al.,Chem.Res.Toxicol.,2016,12251251
AspinoffprojectfromourLCMS/MSmethodsofPFASanalysis
0
3
6
9
12
11/19/2015 11/24/2015 11/29/2015 12/4/2015 12/9/2015 12/14/2015
Concentrationg
/L
6:2FtS
PFOS
TimelineofSampling(2015)
9
11/19FirstAFFFAddition
11/25Endof1stRoundofAFFF
12/1MoreAFFF
12/3EndofAFFF
11/1911/20
11/2311/25 11/30 12/2 12/6 12/9 12/14
12/1512/3
ErikaHoutzetal,2016ACSpresentation,SanDiego
SamplecollectedAnalyzedbyLCMS/MSforPFASSuiteAnalyzedbyTOPAssayAnalyzedbyLCQTOFMS
TotalPFASlevel(TOPassay)muchhigherthantotalconcentrationoftargetedPFASmeasuredinthewastewater
TargetedPFAS
Total TOPassay
QTOF:ExpandedPFASlibrary
workflow Full Scan TIC, triplicate data set
Inspect Data Quality, RT shift, peak intensity
MPP for compounds Grouping and Alignment,
and blank substraction
FBF (Find By Formula using appropriate parameters), search against PFAS library
Recursive Qual Analysis1. Generate Compound list2. Inspect Chromatography3. Inspect Mass spectrum4. review overall score
Further Identification and confirmation1. Literature search2. Standard comparison3. MS/MS spectra
Adatabasewascompiled,whichcontainedthemassesandelementalformulasforallpotentialAFFFfluorochemicals identifiedinpatents.
105compoundsidentifiedinAFFFsamples:C3C15
Accuratemass
Isotopicpatternmatch
WelldefinedChromatographyPeak
12/3/15CompositeInfluent
12/3/15CompositeEffluent
KatieC.HardingMarjanovic et.al.,Environ.Sci.Technol.2015,49,76667674
SFOWWScreening
Expandtheanalytes listbeyondtheoriginalLCMS/MSmethod:foundprecursors,intermediates,andnewreplacementPFASs:ADONA,EtherPFHxS,KetonePFOS
Helpconfirmationofthesourcesandtransformationpathways
WorkingonCompareWWprofilesfromdifferentsources:MPPanalysis
Previously,wehaveusedLCMS/MSserummethodtoanalyzed12PFAScompoundsincatserumsamples.
PilotprojectforbiologicalsampleQTOFscreening
NormalCatsControl(5)
HTCatsCase(5)
Questions Asked For This Study
1.Exposome ofCatSerum:NonTargetSuspectScreening:Beyondthe12commonPFCAs;
Highproductionvolume(HPV)chemicals,pharmaceuticals,pesticides,polyandperfluoroalkyl substances(PFAS)andflameretardants
2. Aretheredetectableprofiledifferencesbetweenthecontrolandcasegroups?
SerumPrep:asimpleproteinprecipitationmethod
MeOH
Vortex
Centrifuge
Evaporation
Reconstitutionandinstrumentanalysis
the MPP (Mass Profiler Professional) non targeted workflow
Full Scan TIC, triplicate injections
MFE (Molecular Feature Extraction) with appropriate parameters
Export to MPP, Grouping and Alignment: Features aligned across all data sets
Apply Quality Control Filters, Blank subtraction, Recursive analysis
PeakfilterMasstoleranceRTtoleranceIonspecies
Export for Identification: exact mass, isotope pattern, retention time1. Search against databases2. Unknowns: elemental
formula generation
Profiles Comparison MPP1. Group as control and case2. Comparison using volcanic
plot, p 2
Identify features that are up or down regulated between case and control groups
Further Identification and confirmation1. Search Chemspider/Pubmed and/or Merck Index
(search by formula, by accurate mass) 2. MS/MS spectra
Exposome Profile
CatExposome:All10samplesFeature:Mass@RT
~1300afteralignment
Cpd Label Name RT Formula Score Mass488 Cpd48benzoicacid 2.105 C7H6O2 99.14 122.036794 Cpd94allylcaproate 3.449 C9H16O2 99.03 202.121347 Cpd471octen3ol 2.593 C8H16O 99.02 174.125857 Cpd571octen3ol 2.88 C8H16O 98.94 174.1261366 Cpd362,2,6,6tetramethyl4methylideneheptane 6.813 C12H24 98.49 214.192766 Cpd66tertbutylhydroquinone 3.046 C10H14O2 98.48 166.0993763 Cpd761,2benzisothiazol3(2H)one 7.567 C7H5NOS 98.3 151.009235 Cpd352{[2,2dimethyl3(oxiran2ylmethoxy)propoxy]met 2.353 C11H20O4 98.27 216.1364302 Cpd30decene 6.194 C10H20 98.27 186.1623490 Cpd49perfluoro1butanesulfonate 4.028 C4HF9O3S 98.24 299.9508116 Cpd1 ID0273from:Env.ScienceTechnol.,2013,47(10), 3.822 C11H18O 98.21 166.1359713 Cpd7 1,2benzisothiazol3(2H)one 7.201 C7H5NOS 98.18 151.00947 Cpd7 allylcaproate 0.724 C9H16O2 98.16 202.1205
583 Cpd58dibutyladipate 4.585 C14H26O4 98.13 258.1835166 Cpd16dibutyladipate 4.511 C14H26O4 98.06 258.1832493 Cpd491,1,2,2,3,3,4,4,5,5,6,6,6Tridecafluoro1hexanesulfo 5.249 C6HF13O3S 98.06 399.94331266 Cpd1 Lufenuron 6.954 C17H8Cl2F8N2O3 97.98 509.9781130 Cpd13ethenyldecanoate 3.95 C12H22O2 97.97 244.1679139 Cpd13ethenyldecanoate 4.054 C12H22O2 97.96 244.168293 Cpd292,6,8Trimethyl4nonanoneID#204 6.104 C12H24O 97.96 244.20487 Cpd87diisopropyladipate 3.332 C12H22O4 97.89 230.1518771 Cpd77ID0270from:Env.ScienceTechnol.,2013,47(10), 8.28 C35H70O2 97.86 568.5426128 Cpd1 1,2benzisothiazol3(2H)one 3.906 C7H5NOS 97.85 151.00934 Cpd4 butyl2methylprop2enoate 0.579 C8H14O2 97.73 188.1052
191 Cpd19dibutyladipate 4.968 C14H26O4 97.69 258.1834423 Cpd4 DL12hydroxystearicacid 7.205 C18H36O3 97.64 300.2669470 Cpd47palmiticacid 7.794 C16H32O2 97.44 256.2404228 Cpd2 2,2,4trimethyl1[(2methylpropanoyl)oxy]pentan3 5.43 C16H30O4 97.41 286.214867 Cpd67isoeugenol 3.045 C10H12O2 97.4 210.0892288 Cpd28decene 6.079 C10H20 97.15 186.1616213 Cpd2 17alphaEstradiol 5.216 C18H24O2 97.05 318.183734 Cpd731H,1H,2H,2Hperfluorooctanesulfonate(6:2FTS) 5.618 C8H5F13O3S 97.04 427.9748
PFBS
PFHxS
6:2FTS
Foodpreservativeandadditives
Presentinonlyonesample
HTvs.Normal:Profilescomparison
Venndiagram,HTvs.Normal
Normal HT
Volcanoplot:UpvsDownRegulation
UpregulatedentitiesDownregulatedentities
5vs5:verylimitedsamplesize
Profilecomparison:HTvs.Normal
IDBrowserUpregulated,Score>80
Name Formula Mass RT Score commentDuloxetine C18H19NOS 343.1241 5.617 96.76 depressiondrugPFOSA C8H2F17NO2S 498.953 6.568 96.73 PFASPFOSAA C10H4F17NO4S 556.9582 6.236 95.59 PFASCatechol C6H6O2 110.0368 0.905 87.47 phenol/pesticideLleucine C6H13NO2 131.0947 0.619 86.9 dietarysupplement
acetaminophen C8H9NO2 151.0634 1.458 86.75 pharmaceuticaltexanol C12H24O3 216.1722 5.738 86.44 coalescentforlatexpaints
myristicacid C14H28O2 228.2093 7.208 85.83 commonfattyacid
acetophenone C8H8O 120.0569 5.286 85.56commonfragrantketoneused
infragance(S)hydroprene C17H30O2 266.2239 7.274 85.37 insecticide
dodecylmethacrylate C16H30O2 254.2236 7.644 82.19anesterthatmightbeusedin
PCP
Profilecomparison:HTvs.Normal
IDBrowserUpregulated,Score>80
Name Formula Mass RT Score commentDuloxetine C18H19NOS 343.1241 5.617 96.76 depressiondrugPFOSA C8H2F17NO2S 498.953 6.568 96.73 PFASPFOSAA C10H4F17NO4S 556.9582 6.236 95.59 PFASCatechol C6H6O2 110.0368 0.905 87.47 phenol/pesticideLleucine C6H13NO2 131.0947 0.619 86.9 dietarysupplement
acetaminophen C8H9NO2 151.0634 1.458 86.75 pharmaceuticaltexanol C12H24O3 216.1722 5.738 86.44 coalescentforlatexpaints
myristicacid C14H28O2 228.2093 7.208 85.83 commonfattyacid
acetophenone C8H8O 120.0569 5.286 85.56commonfragrantketoneused
infragance(S)hydroprene C17H30O2 266.2239 7.274 85.37 insecticide
dodecylmethacrylate C16H30O2 254.2236 7.644 82.19anesterthatmightbeusedin
PCP