Desorption Electrospray Ionization (DESI)

Resolution Sensitivity Time ApplicationSamp. Prep Ambient

AP-MALDI +++ +++ +++ small molec No Yes

DESI +++ +++ ++ small molec No Yes

NIMS ++++ ++++ +++ small molec/prot Yes No

MALDI +++ ++++ +++ small molec/prot Yes No

SIMS +++++ ++++ +++ atoms/small molec Yes No

•AP-MALDI, Atmospheric pressure MALDI•DESI, Desorption electrospray ionization•NIMS, Nanostructure-initiator mass spectrometry•MALDI, Matrix assisted laser desorption/ionization•SIMS, Secondary-ion mass spectrometry

Evolução da Espectrometria de MassasDESI (2004) & The Orbitrap (2005)

Imaging Desorption ElectrosprayIonization (DESI) Mass Spectrometry

HV power supplySolvent

N2

α

β

Nebulizercapillary

Spraycapillary

Gas jet

Spray

Surface Sample

Desorbed ions

Mass spectrometerinlet

Solvent

N2

α

β

Nebulizercapillary

Spraycapillary

Gas jet

Spray

Surface Sample

Desorbed ions

Mass spectrometerinlet

Takats et. al. Science 2004; 306:471.

x, y -moving stage

DESI emitter

Nitrogen inlet

Extended ion transfer tube

y x

360o rotationalstage

9:1 MeOH:H2O (v/v)

1:1 MeOH:H2O (v/v)

1:9 MeOH:H2O (v/v)

Flow-rate and Solvent Effects 1s 1s 1s 1s 1s 200µm/s 10s

d1= 2mmα= 56o HV= 5kVN2= 100 psi Flow: 0.7L/minSpray Cap.: 50/150µm (ID/OD) Neb. Cap.: 250/350µm (ID/OD)

(α

d1

TeeJet Water Sensitive Paperby Spraying Systems Co., Wheaton, IL

4.0

2.0

1.0

0.5

4.0

2.0

1.0

0.5

4.0

2.0

1.0

0.5

Flow-rate (µL/min)

~218µm 21.4 SD

tumor non-tumortumor non-tumor

m/z 885 m/z 788

m/z 448m/z 448m/z 303 m/z 498

m/z 862

a

fed

cb

Selected molecular ion images

H & E stained liver tissue

Wiseman, Poulitaival, Caprioli, Cooks, unpublished.

0

5

10

15

20

25

150 350 550 750 950

m/z

Inte

nsity

-1

0

1

2

3

4

5

6

7

8

9

10

TumorNon-tumor

0

5

10

15

20

25

400 450 500 550 600

m/z

Inten

sity

-1

0

1

2

3

4

5

6

7

8

9

10

TumorNon-tumor

448.3464.3

498.3514.3

Liver adenocarcinoma

Physical Information(based on imaging DESI)

Chemical Information

Analysis of Latent Fingerprints by Imaging DesorptionElectrospray Ionization (DESI) - Mass Spectrometry.

ID

200 400m/z

0

20

40

60

80

100

Rel

ativ

e A

bund

ance

245.17

173.17

311.25

Drugs? Explosives?

IMAGING DESI MS

Map of ∆9-THC (m/z 313) on Latent Fingerprint on Glass

100 150 200 250 300 350 400 450 500 550 600

m/z

0

10

20

30

40

50

60

70

80

90

100

Rel

ativ

e A

bund

ance

400.9

313.3

255.3372.9

325.3

283.3 345.3

356.7227.3157.2 173.2123.2 241.3

297.2 381.0199.3 271.0

-10 µL of .5µg/µL (∆9-THC)-Smear using a pipette tip-Dry 3 min-Blot on glass-Area ~ 300 mm2

-Concentration: ~16ng/mm2

Full Scan (-)

(α

d1

d2)β

d1= 2mm d2= 1mm α = 58o β ~ 10o

HV= 5kV N2= 100 psi

Solvent= MeOH:0.1% NH4OH (9:1 v/v)Flow= 1.5µL/min

-10 µL of .5µg/µL (∆9-THC)-Smear using a pipette tip-Dry 3 min-Blot on glass-Area ~ 300 mm2

-Concentration: ~16ng/mm2

d1= 2mm d2= 1mm α = 58o β ~ 10o

HV= 5kV N2= 100 psi

Solvent= MeOH:0.1% NH4OH (9:1 v/v)Flow= 1.5µL/min

(α

d1

d2)β

Map of ∆9-THC (m/z 313) on Latent Fingerprint on Glass

2mm 2mm

BA

F

CDE

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500

m/z

Rela

tive

Inte

nsity

1- Palmiticacid (C16:0), m/ z2552- cis-Hexadec-6-enoic acid (C16:1), m/ z 2533- Stearicacid (C18:0), m/ z 2833- cis-Octadec-8-enoic acid (C18:1), m/ z 2815- Pentadecanoicacid (C15:0), m/ z 2416- Cocaine , m/ z 3048-∆9-THC, m/ z 3119,10- RDX-Cl35/37, m/ z257,259 9

5 7

8

2

1

7

10

B

A

F

C

D

E

240 250 260 270 280 290 300 310 320

m/z

Rela

tive

Inte

nsity

34

Ion Mode (+) Ion Mode (-)

2

A CB

D E Fm/z 253 m/z 313m/z 304

m/z 304m/z 257 m/z 253

glass

paper tape

paperglass

tape

EASI Source

A) position adjustment nods (3-D variation)

B) nebulizinggas tubing (N2)

C) LCD camera

D) sonic spray device

E) light source

F) cone orifice

G) stainless steel sonic spray capillary

H) glass sample tip

I) fused silica capillary

J) syringe connector

EASI-MS

DESI-MS

Ultrafast EASI-MS Biodiesel Typification

EASI-MS “Cork-Typification” of Wine

Merlot

Cabernet Sauvignon

Carmenére

Chiral Resolution by CSP-Free MS ??

Ion Mobility Mass Spectrometry

David Clemmer

The Ultimate case : Chiralchromatography by MS??

Gas-Phase chiral Cromatography !!!

ChiralChiral Separation By Ion Mobility SpectrometrySeparation By Ion Mobility SpectrometryHerbert H. Hill JrHerbert H. Hill Jr11., ., PrabhaPrabha DwivediDwivedi11, and , and ChingChing WuWu2 2

11 Department of Chemistry & Center for Multiphase Environmental RDepartment of Chemistry & Center for Multiphase Environmental Research, Washington State University, esearch, Washington State University, Pullman, WA 99164Pullman, WA 99164

22 ExcellimsExcellims Corporation; 6 Westside Drive; Acton, MA 01720Corporation; 6 Westside Drive; Acton, MA 01720

Chiral Gas !! R/S 2-Butanol

Department of Chemistry

Hill Research Group Ion Mobility

Spectrometry

1 4 0

1 2 0

1 0 0

8 0

6 0

4 0

2 0

0

-2 0

IMS

Res

pons

e

3 53 02 52 01 5D r i f t T im e ( m s )

1 0 0

8 0

6 0

4 0

2 0

0

-2 0

IMS

Res

pons

e

3 53 02 52 01 5D r i f t T im e (m s )

Mixture of S- and R-Atenolol with drift times of 24.66ms and 25.06ms respectively

S- and R-Atenolol with drift times of 24.61ms and 25.04ms respectively

(R)-(+)-AtenololO

NH2

O(R)

HOHN

(S)-(-)-AtenololO

NH2

O(S)

HOHN

Clowers BH, Hill HH Jr. Influence of cation adduction on the separation characteristics of flavonoid diglycosides isomers using dual-gate

ion mobility-quadrupole ion trap mass spectrometry. Journal of Mass Spectrometry 2006; 41: 339–351

100 Pesticidas em uma única analise

Dichlorvoscis-Mevinphostrans-MevinphosEthoprophosChlorprophamCadusafosPhorateα-HCHHexachlorobenzeneDimethoateCarbofuranβ-HCHDimethipinQuintozeneLindaneDiazinoneChlorothalonilPentachloroanilinePhosphamidonChlorpyriphos-methylVinclozolinParathion-methyl AlachlorCarbarylHeptachlor

Pirimiphos-methylMPCPSMalathionChlorpyriphosAldrinFenthionParathionFenpropimorfTriadimefonOxychlordaneFipronilHeptachlor epoxideChlorfenvinphosPhenthoateQuinalphosTriadimenolMethidathionα-ChlordaneTetrachlorvinphosEndosulfan αγ-ChlordaneProthiofosProfenofosp,p'-DDE

Dieldrino,p'-TDE Cinerin IEndrinNitrofenEndosulfan βFensulfothionp,p'-TDE o,p'-DDT EthionJasmolin ITriazophosCarbophenothionEndosulfan sulphatep,p'-DDTPropargitePiperonylbutoxidePyrethrin IPhosmetBromopropylateFenpropathrinTetradifonPhosalonePhenothrin

Azinphos-methylCinerin-IIλ-CyhalothrinAzinphos-ethylAcrinathrinJasmolin IIcis-Permethrintrans-PermethrinProchlorazCyfluthrin ICyfluthrin IICyfluthrin IIICyfluthrin IVCypermethrin ICypermethrin IICypermethrin IIIFlucythrinate ICypermethrin IVEtofenproxFlucythrinate IIFenvalerate IFluvalinateFenvalerate IIDeltamethrin

• MRM de 14 canais

Total Ion Chromatogram (TIC)

Dic

hlor

v os

Del

tam

e thr

i n

100 pesticide residues84 MRM transitions10ms dwell time

Deltamethrinr2 = 0.998

Pirimiphos-methylr2 = 0.999

Linearity (10-1000pg)

Compound name: DeltamethrinCorrelation coefficient: r = 0.998918, r^2 = 0.997838Calibration curve: 14.0112 * x + 125.819Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

pg/µl0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500

Res

pons

e

0

1000

2000

3000

4000

5000

6000

7000

Tetradifonr2 = 0.999

Compound name: Pirimiphos-methylCorrelation coefficient: r = 0.999511, r^2 = 0.999022Calibration curve: 20.2691 * x + -27.0132Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

pg/µl0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500

Res

pons

e

0

1000

2000

3000

4000

5000

6000

7000

8000

9000

10000

Dichlorvosr2 = 0.998

Compound name: TetradifonCorrelation coefficient: r = 0.999367, r^2 = 0.998735Calibration curve: 6.93326 * x + 21.1316Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

pg/µl0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500

Res

pons

e

0

500

1000

1500

2000

2500

3000

3500

Compound name: DichlorvosCorrelation coefficient: r = 0.998799, r^2 = 0.997600Calibration curve: 12.7295 * x + -2.86235Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None

pg/µl0 25 50 75 100 125 150 175 200 225 250 275 300 325 350 375 400 425 450 475 500

Res

pons

e

0

500

1000

1500

2000

2500

3000

3500

4000

4500

5000

5500

6000

6.000 8.000 10.000 12.000 14.000 16.000 18.000 20.000 22.000 24.000 26.000 28.000 30.000 32.000rt0

100

%

22.4962026

10.204552

9.903516

8.761379

11.321686 19.260

163812.689850

15.0491133 17.451

1421

21.7791940

22.7632058

25.2652358

25.4402379

Scan EI+ TIC

2.00e7RT

Scan

G310704

Selectivity in Baby Food

6.000 8.000 10.000 12.000 14.000 16.000 18.000 20.000 22.000 24.000 26.000 28.000 30.000 32.000rt0

100

%

23.8412355

17.2951570

17.2361563

15.5101356

15.0761304

16.3021451

21.1732035

18.9131764

18.8631758

20.4221945

19.7301862

22.5322198

25.9512608

24.2252401

25.7262581

28.8612957

28.6692934

28.4942913

26.2592645

26.5512680

29.0192976

29.4613029

Scan EI+ TIC

2.00e7RT

Scan

C300708

Strawberry 10mg matrix

Baby Food 1mg matrix

Petroleoma-MS

N

NH

NHN

N

NN

N

RuCl

N

N

NN

RuCl

N

NN

N

RuCl

N

NN

N

RuCl

N

N

NN

+

+

+

+V) a,b

Ink analysis.

Ifa, Gumaelius, Eberlin, Manicke, Cooks. Analyst 2007 ;132: 461.

Seleção doMicrorganismo

Isolamento doMicrorganismo

Ensaio deAntagonismo

Sistema deCLAE/EM

Monitoramento deMetabólitos Bioativos

Derreplicação on-line

Ensaio de Antibiose

Ensaio deBioautografia

Quimiotaxonomiaem CCD

TLC/EASI/ESI

Espectro de massas

Screening AntimicobacterianoTuberculose

637 638 639 640 641 642 643

639.1547

638.1765

640.1635

641.1729

O

OOH OH

CO2MeO

O OHOH

CO2Me

C32H30O14

a)

b)

289

290

247

248

289

247

Bioautografia – TLC-MALDI/ToF

Teste de bioautografia bactéria Rodococcus equi e fungo Cladosporium cladosporióidespara frações simplificadas do extrato bruto do jatobá

250 300 350 400 450 500 550 600 650 700 750 800 850 900m/z0

100

%

2.26e3749.2499

669.1895

479.2776

393.3551 553.1537

781.2461

811.2388

1

2

34

5

6

1''2''

3'' 4''

1'

2'3'

4'

5' 6'

7'

8'

9'

10'11'

R3

R5

OH

OH O

O

R2

O

HO OH

H

R4

R1

OO

-O OHCHO

OO

OH

OH

[M-H]-

-O OHCHO

OO

O OOH

OH

-O OHCHO

OCH3OH-

- 172 -32

m/z 479 m/z 307 m/z 275

m/z200 300 400 500 600 700 800 900

%

0

100 1.16e5479

465

435 493615 762

Espectrometria de Massas Seqüencial

HO

O O

O

OH

OH OH O

OH

OAc

AcO

Isolamento Guiado por Bioensaios

Acetilcolinesterase (AChE)

1 2 3 4 5 6 7 8 9 10 11

ABCDEFGHIJKL

21 14 08 22 18 38 3104

29

2843

01

36

2533

4647

3745

23

32

24

Rolfsi

07

5150

13

051441

15 184048 54

39

09

35

1017 203853

3B

Atividade Fitotóxica