PO-CON1533E

Structural analysis of isomeric chemicalsby using high resolution MS/MS onCurved Field Re�ectron incorporatedwith a novel focusing technology

ASMS 2015 ThP 273

Shuuichi Nakaya1; Keisuke Shima1; Yuzo Yamazaki1

1Shimadzu Corporation, Kyoto, JAPAN.

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Structural analysis of isomeric chemicals by using highresolution MS/MS on Curved Field Re�ectron incorporated with a novel focusing technology

Overview

IntroductionWhile resolution of MS has been greatly improved recently, no matter how high resolution of MS is achieved, MS/MS is essential to conduct analysis of isomeric chemicals, and the analysis is thought to be still signi�cant task given to mass spectrometry. To date, we applied PSD and high energy CID in curved �eld re�ectron to the analysis, since intensities of

fragment ions in the re�ectron were susceptible to isomeric forms. Recently, we incorporated a new focusing technology into the curved �eld re�ectron, which enabled to obtain 10,000 resolution of MS/MS. We will apply the new curved �eld re�ectron toward ambiguous assignment and differentiation of isomeric chemicals.

Isomeric oligosaccharides : Lacto-N-neotetraose(LNnT) Galβ1-3GlcNAcβ1-3Galβ1-4Glc

Lacto-N-tetraose(LNT) Galβ1-4GlcNAcβ1-3Galβ1-4Glc

Isomeric prostaglandin : Prostaglandin E2(PGE2) and prostaglandin D2(PGD2)

Matrix : 10 mg/mL of DHB in methanol.

Cation : Li, Na, Cs, Ba.

Instrument : MALDI-7090 (Shimadzu/Kratos)

Measurement : High-energy CID-MS/MS in positive ion mode.

Collision gas : helium

Collision energy : 20 keV (laboratory frame of reference).

• Precise assignment of isomeric chemicals is conducted with a new MALDI-TOF/TOF MS.• Isomeric carbohydrates and prostaglandins are differentiated successfully.• Choice of cation is signi�cant to obtain informative MS/MS spectrum.

Materials

MALDI-tandem TOFMS

Methods

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Structural analysis of isomeric chemicals by using highresolution MS/MS on Curved Field Re�ectron incorporated with a novel focusing technology

Fig.1 Inside view of MALDI-7090

MS/MS resolution is improved with correcting the spatial distribution of ions in ASDF cell. This is achieved by applying a pulsed electrostatic �eld at the point, at which the precursor and fragments are velocity focussed but are spatially resolved.

Principle of Axial Spatial Distribution Focusing (ASDF)

Vacuum MALDIion source

Lineardetector

Curved �eldre�ectron

Re�ectrondetector

ASDF cellIon gate

CID cell

Fig.2 Axial Spatial Distribution Focusing

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Structural analysis of isomeric chemicals by using highresolution MS/MS on Curved Field Re�ectron incorporated with a novel focusing technology

Fig.3 MS/MS of LNT and LNnT

Fig.4 fragment ions of LNT and LNnT

Fig.5 Effect of metal cations in MS/MS (PGE2)

Results

LNnT

LNT

[M+Na]+

[M+Na]+

O

O

O

O

O

O

O

OH

NHAc

OH OH

OH

HOH2C

HO

HO

HO

HOH2C HO

HOH2C HOH2C

HO

O

HO

O

O

O

O

O

OH NHAc

OH OH

OHHOH 2C

HO

HO

O

HOH2C HO

HOH2C HOH2C

HO

LNnT LNT

185.0420

B1

568.1488 Y3

388.1214

B2

365.1054 Y2

550.1742

B3

203.0526 Y1

568.1848 C3

406.1320 C2

203.0526 C1

208.0580 BZ

185.0420

B1

568.1488 Y3

388.1214

B2

365.1054 Y2

550.1742

B3

203.0526 Y1

568.1848 C3

406.1320 C2

203.0526 C1

0,2 X3

305.0843 0,2 A2

448.1425

3,5 A1 1,4 X2

331.1000

3,5 A1 Y2

259.0788

[M+Cs]+ [M+Li]+ [M+Ba-H]+

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Structural analysis of isomeric chemicals by using highresolution MS/MS on Curved Field Re�ectron incorporated with a novel focusing technology

Fig.6 MS/MS of PGD2 and PGE2

PGE2

PGD2

[M+Ba-H]+

[M+Ba-H]+

PGE2

PGD2

318.9923

Structural analysis of isomeric chemicals by using highresolution MS/MS on Curved Field Re�ectron incorporated with a novel focusing technology

For Research Use Only. Not for use in diagnostic procedures.The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice.

© Shimadzu Corporation, 2015

First Edition: May, 2015

www.shimadzu.com/an/

Fig.7 Conceivable pathways for fragmentations of PGD2 and PGE2, including reported one in ref(1).

Conclusions• High MS/MS resolution achieved by ASDF enables to perform precise assignment in the analysis of isomeric chemicals. • Differentiation of linkage formations between Gal and GlcNAc was conducted successfully by MS/MS of sodium

adducted carbohydrates.• Charge remote fragmentation of bariated PGD2/PGE2 was applied to a differentiation of the isomeric chemicals.• Choice of cation was signi�cant to obtain interpretable fragmentation as well as FAB-tandem MS analysis.

References(1) Zirrolli, J.A., et al; J Am Soc Mass Spectrom., 1990, 1(4), 325-35.(2) Yamagaki, T., et al; J Mass Spectrom., 2006, 41(4),454-62.

342.9906 318.9906

208.9174

154.9069

363.0168

345.0063

PGD2 [M+Ba-H]+ C20H31BaO5 Exact Mass: 489.1218

PGE2 [M+Ba-H]+ C20H31BaO5 Exact Mass: 489.1218

+H

-H

262.9644 -H

+H

-H2O

417.0274

399.0168

-H

-H2O

Or,

304.9761

401.0325

389.0325 +H

Common fragment ions

401.0325

389.0325 +H