Structures for Lossless Ion Manipulations (SLIM ) traveling wave … · 2006. 2. 17. · SLIM SUPER TW IM-MS platform enhances resolution in ion mobility separations of lipid species,

Structures for Lossless Ion Manipulations (SLIM) traveling wave SUPER high resolution

IM-MS for lipidomics

Biological Sciences Division, Pacific Northwest National Laboratory

R. Wojcik, I. K. Webb, Y. M. Ibrahim, J. E. Kyle, K. J. Bloodsworth, S. Garimella, A. Hamid, E. S. Baker, R. D. Smith

LIPIDOMICS – complexity due to structural diversityLipid Maps database: >40,000 entries owing to progress in:

- Front end chromatographic separations- High resolution mass spectrometry- Fragmentation and ion-molecule reaction techniques for structural elucidation

2Shevchenko et al, 2010, Nature Reviews Molecular Cell Biology, 11, 593-598

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LIPIDOMICS – ongoing analytical challenges

Han et al, 2016, Progress in Lipid Research, 61, 83–108J.P. Koelmel et al, 2017, Biochimica et Biophysica Acta, 2017, in pressHankock et al, 2017, Analytical Biochemistry, 524, 45-55

• Improper lipid annotations

• Isobaric/isotopic interferences

• No universal method for structural elucidation

• Low peak capacity; low throughput of front-end separations

IM-MS for lipid separation and structural elucidation

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• Ions move in an electric field through a drift path and interact with a buffer gas • Fast separations, precise drift times• Distinct collisional cross section (CCS) trend lines for lipid classes • Separation of isomeric/isobaric lipids feasible

Kliman et al, 2011, Biochim. Biophys. Acta, 1811, 935–945 ; Kyle et al, 2016, Analyst, 141, 1649–1659

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Challenges for IM-MS lipid analyses

Kyle et al, 2016, Analyst, 141, 1649–1659

• Limited peak capacity• Insufficient resolution of isomers/isobars• Co-elution even with front-end LC

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Resolution in IM-MS

• Resolution between ions depends on differences in their mobilities

• Selectivity – type of gas, dopants

• Physical parameters – temperature, pressure

• Instrumental factors – path length L and E profile across L

• E profile differs among IM MS techniques

• Resolution ~ 𝐿𝐿

7

High V

Low V

DC guard DC guard

DC guard DC guard

RF RF RF RF

RF RF RF RF

Ion confinement for IM-MS in Structures for Lossless Ion Manipulations (SLIM)

y

x

8

y z

x

High VLow V

Traveling Waves (TW) in SLIM for ion transport in IM-MS

TW

TW

TW

xz

TW A

mpl

itude

(V)

Hamid et al, 2015, Anal. Chem., 87, 11301−11308

SLIM TW Serpentine Ultra-long Path with Extended Routing (SUPER) High Resolution IM-MS

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• Lossless transmission - straight paths and 90° turns

• Serpentine path for TW IM-MS

Multi-fold resolution enhancement vs DT IM-MS

• Multi-pass SLIM SUPER IM with expandable path length

Resolution increases with square root of path length

Hamid et al, 2015, Anal. Chem., 87, 11301−11308Deng et al, 2016, Anal. Chem., 88 (18), pp 8957–8964Wojcik et al, 2017, Int. J. Mol. Sci., 18, 183Deng et al, 2017, Anal. Chem.,89 (8), pp 4628–4634

TW SLIM SUPER IM-MS platform

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+ 1.3 m = ~ 16 m - full mobility range

SUPER long path ~ 15 m x number of passes + 1.3 m - selected mobility range

pass

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1. PC16:1(9Z)/16:1(9Z) 2. PC16:1(9E)/16:1(9E)

phosphocholine fatty acid cis/trans isomerswith 16 meter SLIM IM-MS

1 2

DT IM-MS 16 m SLIM TW IM-MS

400 410 420 4300

1

Nor

mal

ized

inte

nsity

Drift time (ms)

1

2

34 36 38 400

1

Nor

mal

ized

Inte

nsity

Drift time (ms)

(M+H)+

Phosphocholine fatty acid double bond positional isomers with 16 meter SLIM IM-MS

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1. PC18:1(6Z)/18:1(6Z) 2. PC18:1(9Z)/18:1(9Z)

1 2

16 m SLIM TW IM-MSDT IM-MS

36 38 40 420

1

Nor

mal

ized

inte

nsity

Drift time (ms)460 480 500

0

1

Drift time (ms)

Nor

mal

ized

inte

nsity

1

2

(M+H)+

Ganglioside (head group) isomerswith 16 meter SLIM IM-MS

13

1,2

(M-2H)2-1: GD1b (36:1) 2: GD1a (36:1)

DT IM-MS 16 m SLIM TW IM-MS

1,2 - mixture

36 40 440

1

Nor

mal

ized

inte

nsity

Drift time (ms)300 320 340 3600

1

Nor

mal

ized

inte

nsity

Drift time (ms)

1 3 - mixture

SLIM IM-MS – brain polar lipid extractNegative ion mode

m/z

m/z

1.3 meter SLIM 16 meter SLIM

Drift time (ms) Drift time (ms)

SLIM IM-MS – brain polar lipid extractganglioside isomers (Gd1a, Gd1b)

m/z

Drift time (ms)

m/z

Drift time (ms)

((GD1b (36/2) ((GD1a (36/2)

1.3 meter SLIM 16 meter SLIM

(M-2H)2-

Drift time (ms)

m/z

Drift time (ms)

SLIM IM-MS – brain polar lipid extract, glycerophospholipids

m/z

1.3 m SLIM 16 m SLIM

PC(16:0/20:3)_A/B/CPC(18:1/18:2)(M-CH3)-

SLIM IM-MS – brain polar lipid extract, glycerophospholipids

Drift time (ms)

m/z

PG(16:0/18:1) ?PA(18:0/22:6) ?

Drift time (ms)

m/z

Drift time (ms)

1.3 meter SLIM 16 meter SLIM

1.3 m SLIM 16 m SLIM

Multi-pass TW-SLIM SUPER IM-MS of lipid isomers

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PC (16:1(9Z)/16:1(9Z) – 275.3 Å2PC (16:1(9E)/16:1(9E) – 276.7 Å2

0 50 100 150 200 250 300

1.0

1.5

2.0

2.5

3.0

3.5re

solu

tion

path length (meters)

PresenterPresentation Notes

Summary

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• SLIM SUPER TW IM-MS platform enhances resolution in ion mobility separations of lipid species, isomers

• SLIM SUPER IM-MS has broad applicability for separation and structural characterization of lipids in mixtures

Future directions• Ultrasensitive and ultrahigh resolution SUPER SLIM (R.D. Smith, WP 346)• Compression Ratio Ion Mobility Programming (CRIMP) (S. Garimella, WP 349)• CRIMP for greater sensitivity and dynamic range of SLIM SUPER IM-MS

(L. Deng, ThOG 3:30 pm)• 3 dimensional (multi-level) SLIM to increase path lengths and peak capacities for

full mobility range analyses (A. Hamid, ThP 400)

Y.M. Ibrahim et al., 2017, Anal. Chem., 89 (3), 1972–1977

Acknowledgements

National Institute of General Medical SciencesBiomedical Technology Research Resource at PNNL

DOE Office of Biological and Environmental Research

PNNL SLIM and IM-MS team

Lipid team: Jennifer E. Kyle, Kent J. Bloodsworth

Structures for Lossless Ion Manipulations (SLIM) traveling wave SUPER high resolution �IM-MS for lipidomicsLIPIDOMICS – complexity due to structural diversityLIPIDOMICS – ongoing analytical challengesIM-MS for lipid separation and structural elucidationChallenges for IM-MS lipid analysesResolution in IM-MS Ion confinement for IM-MS in �Structures for Lossless Ion Manipulations (SLIM)Traveling Waves (TW) in SLIM for ion transport in IM-MS SLIM TW Serpentine Ultra-long Path with Extended Routing (SUPER) High Resolution IM-MSTW SLIM SUPER IM-MS platform�phosphocholine fatty acid cis/trans isomers�with 16 meter SLIM IM-MS �Phosphocholine fatty acid double bond positional isomers with 16 meter SLIM IM-MS Ganglioside (head group) isomers�with 16 meter SLIM IM-MS SLIM IM-MS – brain polar lipid extractSLIM IM-MS – brain polar lipid extract�ganglioside isomers (Gd1a, Gd1b)SLIM IM-MS – brain polar lipid extract, �glycerophospholipidsSLIM IM-MS – brain polar lipid extract, �glycerophospholipidsMulti-pass TW-SLIM SUPER IM-MS of lipid isomersSummaryAcknowledgements