L.RamosGrupodeQuímicaAmbiental

Dpto.AnálisisInstrumentalyQuímicaAmbiental

e-mail:l.ramos@iqog.csic.es

¿QUÉPUEDEHACERLAQUÍMICAPORELMEDIOAMBIENTE?

Elpapeldelaquímicaanalítica

VICurso deDivulgación.LosAvancesdelaQuímicaysuImpactoenlaSociedad

LaOMSrelaciona13millonesdemuertesenelmundoconfactoresambientales

Lacontaminación esunproceso globalquenoentiendedefronteras

Losnuevosusoscreannuevasfuentespotencialesdecontaminantes

WhyChemistry?Throughouthumanhistory,chemistryhashelpedussurvive.Turningstonesintotools,makingfire,creatingceramics,andpreservingfoodweresomeoftheearliestbenefitsofchemistry

Herramientasparalaproteccióndelmedioambienteyelhombre

Legislación

Monitorización

Evaluación

Contaminantes Orgánicos Persistentes (COPs)PersistentOrganicPollutants(POPs)

• Adoptedon22May2001;enteredintoforceon17May2004

• Compoundsthatoncereleasedintheenvironment,• Remainforaexceptionallylongtime:persistent• Accumulateinthefattytissueoflivingorganismsincludinghumans:lipophilic

• Widelydistributedthroughouttheenvironment:long-rangetransport

• Toxic tobothhumansandwildlife

DieldrinAldrin Chlordane DDT

Endrin Heptachlor HCBMirex

ToxaphenePCDD PCDFPCB

The“dirtydozen”

Otherchlorinatedcompounds

Otherbrominatedcompounds

26chemicalsorgroupsofchemicalsareregulatedasPOPsundertheStockholmConvention

Dataanalysisand

Interpretation- Unambiguousidentification- Quantitation

InstrumentalAnalysis

Chromatographicseparation- Detection

SamplePreparation- Extraction- Purification- Fractionation

Environmentaltraceanalysis:POPsChallenges–>Theanalyticalprocedure

• Lowconcentrations• Complexmatrices• Manyclasses

• Numerousfamilies:notseparatedonasinglecolumn;analysison2phases

• Lowconcentrations

Instrumentalanalysis:GasChromatography(GC)

J.Sep.Sci.27(2004)595

Analysis ofPCBs byGC-µECD

JCA1152(2007)254

The risk ofinterference increases asthe complexity ofthematrix increases andasthe initial sample size decreases

…andasthe sample preparation procedure is simplified

GC-MS(SIM) canaddtherequiredselectivity,but…

…theninformation regardingotheranalytesislost

IsGC´GC necessary?

GC´GC:theconcept

GC´GC:theconcept

MatchbetweenseparatorandsampledimensionalityinGC´GC

GC´GC schematicdiagram

Injector

1D 2DModulator

Detector

1tR

PM

Y

X

2tRY2tRX

X+Y

0

1

2

3

4

5

6

7

8

Injector

1D 2DModulator

Detector

1tR

PM

Y

X

2tRY2tRX

X+Y

0

1

2

3

4

5

6

7

8

Standard (10-60m,0.25mm,0.5µm)

Non-polar (DB-1,DB-5)

1D FastGC(1-2m,0.10-0.18mm,0.10-0.18µm)

Polarorshapeselective(DB-1701,Rtx-PCB)

2D

?

GC´GC schematicdiagram

Injector

1D 2DModulator

Detector

1tR

PM

Y

X

2tRY2tRX

X+Y

0

1

2

3

4

5

6

7

8

Injector

1D 2DModulator

Detector

1tR

PM

Y

X

2tRY2tRX

X+Y

0

1

2

3

4

5

6

7

8

PM

2tRX 2tRY

2tRX

2tRX2tRX

2tRX

2tRX

2tRY

2tRY2tRY

2tRY

2tRY

1 2 3 4 5 6 7

Inte

nsity

1tR, 2tR

PM

2tRX 2tRY

2tRX

2tRX2tRX

2tRX

2tRX

2tRY

2tRY2tRY

2tRY

2tRY

1 2 3 4 5 6 7

Inte

nsity

1tR, 2tR

Rawdatasignalasrecordedbythedetectorthroughtheentireseparationprocess

1tR

2tR

Intensity

YX

XY

1tRX1tRY

2tRY2tRX

7654321

1tR

2tR

1tR

2tR

Intensity

YX

XYY

1tRX1tRY

2tRY2tRX

7654321

1tR

2tR

Reconstructionofthetwo-dimensionalcontour-plotfromthecollectedhigh-speedsecondarychromatograms.Similarsignalintensitiesareconnectedbycontourlines

TypicalGC´GCset-up

Quad-jetdual-stagecryogenicmodulator

2Dovenwiththecoiled2Dcolumn

Peakmodulation

• Originalpeakis“chopped”into3-5peaks

• Sensitivityenhancementoccursthroughfocusing

• 2Dpeaksareonly200-400ms wide

• Needdetectorcapableofdefiningpeaks

- Hundredsofspectra/sec

1545

GC´GC features

• Higherpeakcapacity:morechromatographicpeaksperspace• Eliminatestheneedforsecondcolumnconfirmation

- (Allow)multipleclassesinthesamerun- (May)eliminateneedforextractfractionation

• Fast analysis- Requiresfastdetector:FID,µECD,ToF MS

• Providesstructuredchromatogramsforexcellentselectivity• Improvedsensitivity

Structuredchromatograms

HP-1´ HT-8

JCA958(2002)203

GC´GC−ToF MS

Twocomplementaryoptionstoachieveseparationofco-elutingpeaks

GCxGC ToF MS

Co -elution

Peak capacity Deconvolution

Separation

Analyticalresolution

Chromatographicresolution

GCxGC

Co -elution

Peak capacity Deconvolution

Separation

Analyticalresolution

Chromatographicresolution

AnalyticalinformationfromGC´GC−ToF MS

Targeted

Specificanalytes-Coelutions-Matrixeffect

Quantification(trace,ultra-trace

level)

Untargeted

Screening-Multi-classanalysis-Non-targetknowns

Identification- Unknownknowns

- Unknowns

TargetanalysisChallengingseparations:Pesticideanalysis

Rtx-1´ Rtx-200

JCA1186(2008)202

BaA,MW,228

Chr,MW,228

CCP,MW,226

TargetanalysisChallengingseparations:EUPAHsinsediments

ABC393(2009)1697

DB17-MS

DB5-MS

Optimad-6

TargetanalysisChallengingseparations:EUPAHsinsediments

1

2

3

5

4

7

6 8

9

1011

12

13

14

15

16 DB-5´ BPX-50

BaA (2),Chr(4);MW,228CCP(3);MW,226BbF (6),BjF (7),BkF(8);MW,252IcdP (10);MW,276BghiP (11);278

Untargetedanalysis:ScreeningMulti-classanalysis:Organohalogenated compounds

0 20 40 60 80 100 120 140 1600

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title0 20 40 60 80 100 120 140 160

0

1

2

3

4

5

6

Y Axis

Title

X axis title

PCBsPCDD/FsOCPsPBDEsPCNsCTTsPCTs

HT-8 xBPX-50

1tR (min)

2 tR(m

in)

JCA1186(2008)312

Roof-tilesubclasses:Tentativeidentificationofother PBDEs

Untargetedanalysis:ScreeningMulti-classanalysis:OBsinbluefin tuna:PBDEs

JCA1218(2011)6995

1

23 4

Deca-CB

Nona-CBsOcta-CBs

Tri-BHD

Tetra-BHD

Untargetedanalysis: Identification:UnknownsNaturallyproducedOBsinbluefin tuna

JCA1218(2011)6995

5

Polybrominated hexahydroxanthene derivates (PBHDs)

• Newtri- andtetra-BHDderivates tentativelyassigned

200 250 300 350 400 450 500

100

200

300

400

500

600

700

800

900

1000 305 225

387

466

331

Caliper - sample "0510092:1", 3258 , 4.481 sec , sec to 3258 , 4.481 sec , sec - 3258 , 4.150 sec , sec to 3258 , 4.196 sec , sec

*newpossibletri-BHDisomer

**tri-BHDstandard

Untargetedanalysis:UnknownknownsNewlyidentifiedtri- andtetra-BHDsintuna

Newlyidentifiedtri- andtetra-BHDsintunamuscle

Newlyidentifiedtri- andtetra-BHDisomersfurtherconfirmedby GC-QqQ MS

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

MRM546->385MRM546->387

MRM544->385

A

B

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

MRM546->385MRM546->387

MRM544->385

tetra-BHD

MRM546->385MRM546->387

MRM544->385

A

B

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

MRM546->385MRM546->387

MRM544->385

A

B

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

tri-BHD

MRM466->387MRM466->385MRM468->387

tetra-BHD

MRM546->385MRM546->387

MRM544->385

tetra-BHD

MRM546->385MRM546->387

MRM544->385

A

B

EST48(2014)9591

Untargetedanalysis:UnknownsNewmixedhalogenatedcarbazole

J.Escobar-ArnanzIQOG-CSIC

X.OrtizM.Pena-AbaurreaE.ReinerEnvironment Canada

ASSESSMENTOFTHEORGANOHALOGENATEDBURDENINBIOLOGICALSAMPLESBY

NON-TARGETGC´GC−TOFMS

HT-8 xBPX-50

Untargetedanalysis:UnknownsOrganohalogenatedcompoundsintunamuscle

HT-8 xBPX-50

>5000peaks!

Scripts:Isotopeclustersearch

Number of Cl-

1 2 3 4 5 6

Num

ber o

f Br-

1

2

3

4

Scripts

HT-8xBPX-50

Automaticdatafilteringbyclassificationsandscripts:Tunamuscle

ü 354filteredbythescriptfunction• 269Cl/Brcompounds>50%identifiedbylibrary(similarity>750)

70%

27%

3.2%

Percentdistribution

Cl

Br

Cl-Br

CHARACTERIZATIONOFAROMATICORGANICCOMPOUNDSGENERATEDINANUNCONTROLLED

TIRELANDFILLFIREBYGC´GC−TOFMS

J.Escobar-ArnanzIQOG-CSIC,Madrid

G.BlancoMNCN-CSIC,Madrid

S.MekniE.EljarratD.Barceló

IDAEA-CSIC,Barcelona

Uncontrolledtirefire- FromMay14th toJune2nd,

2016(5dayslatterinMadridarea)

- Only15,000Toftiresremainedunburnt

Usedtires:stockpiledorillegallydumped

Seseña´s dump- ThelargestoneinSpainandoneofthelargestinEU- Usedsince2002- Declaredillegalon2015- Extension,117,000m2

- 70,000-90,000Toftires

Characterizationofaromaticorganiccompoundsinsoils:Uncontrolledcombustionofatirelandfill

• MainfocusonVOCsandPAHs(target)• Steeretal.(1995):PCDD/Fs(target)• Wangetal.(2007):GC-qMS;165PAHs

- EPAPAHsand4-7ringsPAHs>>alkyl-PAHs- S-,O- andN-PAHs

• Downard etal.(2015):GC-qMS- 4-5ringsPAHs- N- andO-PAHs

Seseña´s fire(target)• Nadaletal.(2016):GC-HRMS,LC-FD

- PCBs- PCDD/Fs- EPAPAHs- Selectedtraceelements

Generaloverview:Organohalogenatedcompounds

HT-8xBPX-50

SP-1

Scripts

PAHsandrelatedcompounds:EPAPAHs

74

78

0

4

8

12

16

20

24 SP-1SP-2SP-3SP-6

Normalizedabundances

(Normalizedbyabundanceof13C-BDE28)

2-3fusedrings

4-5fusedrings 6-7fusedrings

Reconstructed2Dbubbleplot

• Normalizedabundances

• SP-5-6:percentcontributionreferred tothatinSP-4

1 2

3

48

5 67

9

10

0

2

4

6

800 1200 1600 2000 2400 2800 3200 3600

2 t R(s

)

0

2

4

6

800 1200 1600 2000 2400 2800 3200 3600

2 t R(s

)

0

2

4

6

800 1200 1600 2000 2400 2800 3200 3600

2 t R(s

)

0246

800 1200 1600 2000 2400 2800 3200 3600

2 t R(s

)

1tR (s)

SP-4(92comp)

SP-5(87comp)

SP-6(46comp)

SP-7(34comp)

Othernon-regulatedPAHs

• 16additionalnon-substitutedPAHs:ü Mostofthem,4to5fusedrings

• 29alkylatedPAHs:ü Soils:mostnumerousclasses:alkyl-

phenanthrene/anthracene(4methyland7dimethyl)ü Ash:methyl-chrysene/benzo[a]anthracene(only

minorcontributorsinsoils)

• 34heterocyclicPAHs

Non-regulatedPAHsandalkylatedPAHs

m/z

242

B

60 70 80 90 100

500

1000

95 76 87 99 91

Caliper - sample "18071604:1-SENA 4-conc", 2892 , 5.200 sec , sec to 2892 , 5.200 sec , sec - 600 , 0.000 sec , sec to 600 , 0.000 sec , sec

50 100 150 200 250 300 350 400

500

1000 242

120 87 163

Peak True - sample "18071604:1-SENA 4-conc", peak 2878, at 2892 , 5.200 sec , sec

50 100 150 200 250 300 350 400

200

400

600

800

1000 167 86

56

108 252

Library Hit - similarity 970, "Chrysene, 1-methyl-"

Comp.#11260 70 80 90 100

500

1000

77 87 81

Caliper - sample "18071604:1-SENA 4-conc", 2892 , 5.360 sec , sec to 2892 , 5.360 sec , sec - 600 , 0.000 sec , sec to 600 , 0.000 sec , sec

50 100 150 200 250 300 350 400

500

1000 254

126

226 150

Peak True - sample "18071604:1-SENA 4-conc", peak 2879, at 2892 , 5.360 sec , sec

50 100 150 200 250 300 350 400

200

400

600

800

1000 139

338 111

75 319 169

Library Hit - similarity 978, "2,2'-Binaphthalene"

Comp.#113

m/z

254

Ash SP-4

Me-chrysene/benzo[a]anthracene

Bynaphthalene

HeterocyclicPAHs

Compoundsidentifiedinsamples

S-PACs;15%

O-PACs;8%

N-PACs;5%

Other;72%

ü 20S-PACs• DBT(/naphtho-thiophene)• 5Me-DBT/naphtho-thiophene• 5di-Me-DBT/naphtho-thiophene• Phenyl-benzothiazole• Benzo[b]naphtho[2,1-d]thiophen

ü 9O-PACs• 9-H-fluoren-9-one• 9,10-anthracenedione• 2-Me-9,10-antracenedione

ü 5-N-PACs• Noazaarenes• Carbazole

S-PACs(20comps)

SP-4

SP-5

SP-6

SP-7

Regulatedandemerginghalogenatedcompounds

üNoPCBs*üNoPCDD/Fs(LOD!)*

üSomeOCPs(p,p´-DDE,HCB,…;apparentlynotrelatedtothefireeven)

üMinoramountsofPBDEsandhalogenatednorbornenesüSeveralOPFRs

ü4non-identifiedhalogen-containingcompounds

* AgreewithNadaletal.Environ.Intern.97(2016)37

OPFRs

Ø s-PLE;LC-QqQMS 3.Resultsasng/gsample

3 Giulivo etal.J.Chromatogr.A1474(2016)71

Analyte SP-1 SP-2 SP-3 SP-4 SP-5 SP-6 SP-7 AshTCEP 5.02 5.66 9.41 8.15 24.7 nq 0.10 7.53TPPO 0.63 0.81 0.94 0.02 0.16 1.13 nd 0.57TCPP 1.67 12.4 3.82 5.39 15.0 6.83 1.99 4.97TDCPP nd nd nd nd 0.09 0.34 0.01 ndTPHP 0.58 0.06 nd nd 0.05 2.28 1.35 ndTBP 4.00 nd 6.19 nd 24.1 3.71 6.70 0.58DCP 0.98 9.50 1.51 nd nd 1.83 nd 3.61TBOEP 2.00 1.15 1.58 nd 3.06 1.35 nd 13.7TCP 4.74 57.7 8.06 nd nd nd nd 61.1EHDP 2.04 5.84 nd 154 1.29 86.1 26.7 474IDPP nd nd nq nd nd nd nd ndIPPP 0.35 0.33 14.1 nd 3.62 nd 12.8 ndTHP 1.69 nd nd nd nd nd nd nqTEHP nq nq nd 3 3.38 nd nd 0.85ΣOPFRs 23.7 93.4 45.6 170 75.5 104 49.6 567

FINALREMARKS

TheEnvironmentalAnalyticalChemistry:

• Evaluatestheefficiencyoflawimplementationthroughmonitoringprograms

• Contributestoearlydetectionofemergingenvironmentalissues

• GC×GC–ToF MSisapowerfulinstrumentaltechnique

• Adapttotheproblemathand• Identificationofnon-targetedandunknowncompounds

Laquímica analítica protegeelmedio ambiente

L.Ramosl.ramos@iqog.csic.es