T-WaveTM: From innovative technology to · ©2011 Waters Corporation 2 Our Mission Develop a Keen Understanding of our customers’ needs Work on Purposeful Innovations to solve analytical

©2011 Waters Corporation 1

T-WaveTM: From innovative technology to purposeful innovation

Ronan O’Malley Senior Manager

MS Product Management


Our Mission

Develop a Keen Understanding of our customers’ needs

Work on Purposeful Innovations to solve analytical challenges and remove bottlenecks

Deliver Meaningful Impact to our customers’ businesses and peoples’ lives


Tandem Quad and Time of Flight LC/MS & MS/MS


ACQUITY UPLC

Our high-performance LC/MS strategy starts with ACQUITY UPLC.

http://www.waters.com/WatersDivision/ContentD.asp?watersit=JDRS-6EMRYQ�


Speed, Sensitivity, Resolution

HPLC-MS/MS

%

Time 2.0 2.5 3.0 1.5 0.5 1.0 0

UPLC-MS/MS


UPLC – The next Challenge

Urine sample drug metabolite study 2x150mm 1.7µm ACQUITY BEH C18

Chinese Ginseng Extract 2x150mm 1.7µm ACQUITY BEH C18


2003 – Introduction of T-Wave


Stacked Ring Ion Guides - Waters’ Implementation

RF Voltages - ~ 400 V pk-pk max

RF Frequency – ~ 1 to 3 MHz range

Pressure - <10-4 to 3 mBar

Length - 10 - 25 cm

RF (-)

RF (+) Electrode Spacing 1.5 mm (centre-to-

centre)

Ion Exit

Aperture Diameter 5.0 mm

Electrode Thickness 0.5 mm

Ion Entry


T-Wave Potential Energy Surface Pot

x y, z,( )

Radial Position r (mm)

Axial Position z (mm)

Effective Potential V* (V)

15

10

5

0

10

20

30

2 1 0

-1 -2


T-Wave Potential Energy Surface

Pot

x y, z,( )

Radial Position r (mm)

Axial Position z (mm)

Effective Potential V* (V)

15

10

5

0

10

20

30

2 1 0

-1 -2

10V Pulse


To reduce ion residence time in gas-filled ion guides, facilitating fast switching experiments

— Launched 2003

o Quattro Premier

Travelling Wave Ion Guide

Printed Circuit Boards

Gas In

Ring Electrodes

Ion TransmissionAperture

EndPlate

SidePlate

©2009 Waters Corporation | COMPANY CONFIDENTIAL

Time1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75

%

0

100

1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75

%

0

100

1.30 1.35 1.40 1.45 1.50 1.55 1.60 1.65 1.70 1.75

%

0

100 6727

7275

7492

High MRM Data Acquisition Rates that could keep up with UPLC

Optimal MS/MS performance

no matter how rapidly you need to acquire data – UPLC compatible…

100ms dwell

10ms dwell

1ms dwell

T-Wave Collision Cell


ScanWave Mode Ion Accumulation

DC Barrier RF Barrier (m/z dependent)

Storage Region Scanwave

To Scanning Quad

Potential Energy

Low m/z Ion

High m/z Ion

Intermediate m/z Ion


ScanWave Mode Ion Transfer

Travelling Wave

Potential Energy

RF Barrier

Low m/z Ion

High m/z Ion



ScanWave Mode Ion Transfer

Potential Energy

DC Barrier RF Barrier

Storage Region

Scanwave

Low m/z Ion

High m/z Ion



ScanWave Mode Ion Ejection

Travelling Wave

Ion Current

Time

Potential Energy

DC Barrier RF Barrier

Low m/z Ion

High m/z Ion




RF Barrier

Travelling Wave

Ion Current

Time

Potential Energy

DC Barrier

Low m/z Ion

High m/z Ion




RF Barrier

Travelling Wave

Ion Current

Time

Potential Energy

DC Barrier

Low m/z Ion

High m/z Ion



ScanWave – High Sensitivity Full Scan MS/MS on a TQ

Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75 3.00

%

0

100S/N:RMS=60.14

m/z50 100 150 200 250 300 350 400 450 500 550 600 650

%0

100195

174

105148

236 397365 448

ScanWave enhanced

Product ion spectrum

UPLC/MS/MS

50fg Reserpine

on column

ScanWave enhanced

Product ion scanning

chromatogram

(m/z 195)


Rapid MS to MRM Switching


m/z100 200 300 400 500 600 700 800 900 1000

%

0

100

m/z100 200 300 400 500 600 700 800 900 1000

%

0

100253.3

123.0

293.3

397.3441.4

619.6485.5

329.3

214.1

100.1149.3 279.3

251.1

415.3

363.4

546.4468.6

600.7

Time0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40

%

0

100

0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40

%

9

0.20 0.40 0.60 0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40

%

2

Targeted compound MRM positive ESI

Fluticasone in plasma

Matrix Profile MS full scan negative ESI

Matrix Profile MS full scan positive ESI

Plasma matrix Spectral information

Qual

itat

ive

Quan

tita

tive


Ele

ctri

c Fi

eld

Diffuse Ion Cloud

Maximising signal

Minimising noise

Wide Ion Tunnel Conjoined to Narrow Ion Tunnel

Off-Axis design


Ring Electrodes Opposite phase of RF applied to adjacent plates

Ele

ctri

c Fi

eld

Diffuse Ion Cloud

Ions In

Ions Out

Compact Ion Cloud

15mm

5mm

~11mm

~120mm

High pressure ion guide design Conjoined Ion Guide


Ions + Gas From API

Source

Rough Pump

Gas

Ions

Differential Aperture

Conjoined Ion Guides

Ion Trajectories

Additional design benefit : Robustness


Conjoined + Second Ion Guide Hardware


Conjoined + Second Ion Guide Hardware

T-Wave


Time0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40

%

0

100

Time0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40

%

0

Xevo TQ

Xevo TQ-S Enhanced sensitivity 30X increase in peak area 30X increase in signal:noise

An increase in sensitivity? Rel

ativ

e io

n a

bundan

ce

Prostaglandin (Plasma)

UPLC/MRM, ESI -


Time0.50 0.60 0.70 0.80 0.90 1.00 1.10 1.20 1.30 1.40

%

0

100

Time0.50 0.60 0.70 0.80 0.90 1.00 1.10 1.20 1.30 1.40

%

0

Xevo TQ

Xevo TQ-S Enhanced sensitivity 129X increase in peak area 25X increase in signal:noise

An increase in sensitivity? Rel

ativ

e io

n a

bundan

ce

Desompressin (Peptide)

UPLC/MRM, ESI +


An increase in sensitivity?

Fenuron ESI+ 30 7Metamitron ESI+ 32 15Acephate ESI+ 27 7Chlortoluron ESI+ 27 8Aldicarb ESI+ 27 6Demeton S Methyl ESI+ 26 9Phoxim ESI+ 64 19Kresoxim Methyl ESI+ 64 4Azinphos Methyl ESI+ 42 6Azoxystrobin ESI+ 45 4Dimethoate ESI+ 23 10Acetamiprid ESI+ 30 28Fluticasone ESI+ 30 3Formoterol ESI+ 39 4Nefadazone ESI+ 28 3Desmopressin ESI+ 129 25Salmeterol ESI+ 41 8Alprazolam ESI+ 21 13Reserpine ESI+ 25 5Ibuprofen ESI- 13 16Prostaglandin E2 ESI- 30 37

Mean Difference 38 11

Relative Peak Area

Relative S:N

Compound NameIonisation

Mode


At the limit Xevo TQ-S

6 replicates

RSD<20%

Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75

%

0

1000.83

Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75

%

0

100

UPLC/MRM of Verapamil

(solvent standard) 400 μL/min UPLC

2fg on column

Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75

%

0

100

0.83

Solvent

Blank

0.1fg on column

220 Zeptomoles on column

(~130,000 molecules)


Time0.50 0.55 0.60 0.65 0.70 0.75 0.80 0.85 0.90 0.95 1.00 1.05 1.10 1.15 1.20 1.25

%

0

100

455 > 165 (Verapamil)2.66e7

0.83

Verapamil, 0.5pg/µL spiked

into supernatant from 2:1 ACN:Plasma protein

precipitation.

10µL injections

ACQUITY BDH 2.1x 50

600µL/min

Plasma injection 1

Plasma injection 2000

Plasma injection 4000

>4000 on column injections

RSD of peak areas < 5%

Assay Robustness –UPLC/MRM ESI + Xevo TQ-S


m/z900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 2100 2200 2300 2400 2500 2600 2700

%

0

100

m/z900 1000 1100 1200 1300 1400 1500 1600 1700 1800 1900 2000 2100 2200 2300 2400 2500 2600 2700

%

0

100

1303.2

1234.7

1173.0

1117.2

1066.51233.8 1302.3

1379.8

1310.3

1954.3

1804.01466.0

1387.2

1675.31563.6

2131.8

ESI Positive Ion Sensitivity –Synapt G2-S

24,000 mw Protein

Synapt G2-S

Synapt G2

Synapt G2

m/z1286 1288 1290 1292 1294 1296 1298 1300 1302 1304 1306 1308 1310 1312 1314 1316 1318 1320 1322 1324

%

0

100 1303.2

1302.31310.2

Synapt G2

Synapt G2-S >10X increase

in signal


UPLC/MSE

Fragment and Molecular ion Information at UPLC rates for every peak in the Chromatogram


m/z100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460

%

0

100Ver0319_002 1179 (8.845) 2: TOF MS ES+

4.15e5

105.0699 177.0913133.0646

165.0916

150.0680

455.2909

303.2072

166.0948

260.1651

185.0778 239.0966261.1644

304.2098

305.2119

456.2942

457.2968

-0.1 mDa

-0.1 mDa

-0.0.mDa

-0.1mDa -0.0 mDa

-0.5mDa

m/z mDa ppm455.2910 -0.10 -0.22303.2073 -0.10 -0.33260.1651 0.00 0.00177.0916 -0.30 -1.69165.0916 0.00 0.00150.0681 -0.10 -0.67105.0704 -0.50 -4.76

UPLC MSE data for verapamil in rat bile , obtained at 10 spectra per second. The excellent exact mass performance combined with MassFragment software allow fragment ions to be assigned structures automatically and compounds to be characterized quickly with high confidence.

UPLC/MSE: Confirm the Identity and Structure of Compounds


SYNAPT G2-S Geometry


TRIWAVE

©2011 Waters Corporation 41 Drift Time

m/z

Travelling Wave Ion Mobility Separation


Maximizing Separation Ion Mobility Separations

Bile Salts

Metabolites


IMS Off

IMS On

Drift time-enhanced precursor-product ion association increases specificity

With Drift Time Alignment 35 product ions tentatively

associated with one precursor

86% ↓ in distraction

Increased search specificity

Without Drift Time Alignment 254 product ions tentatively

associated with one precursor


0

200

400

600

800

1000

1200

1400

1600

1800

2000

1D 2D-3 Fraction 2D-5 Fraction

Prot

eins

LC Method

C.elegans: increased protein IDs with increase in peak capacity (LC+IMS)

1D=1ug; 2D(3)=1.5ug; 2D(5)=2.5ug

<2h <4h <6h

MSE

HDMSE

MSE

HDMSE

80%

280%


ASAP

Atmospheric pressure Sample Analysis Probe

Direct sample analysis — Fast (<1 minute)

— No sample prep

— No chromatography

— Solids and liquids


Scan20 40 60 80 100 120 140 160 180 200

%

0

10069

75

1,2- dinitrobenzene

Drift Time (Bins)

ASAP ion mobility separation 1,2- and 1,3-dinitrobenzene

O-

O-

OO

N+

N+ O

-

O-

O

O

N+

N+

1,3- dinitrobenzene


Unlocking the full power of HDMS ….ASAP analysis of isomers


Unlocking the full power of HDMS ….ASAP analysis of isomers


20

40

60

80

100

120

140

160

180

100 200 300 400 500 600 700 800 900 100040

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

Driftime plot of crude oil components:

ASAP Analysis of Crude Oil


40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

DBE=0DBE=1DBE=6DBE=7DBE=8DBE=9DBE=10DBE=11DBE=12DBE=2DBE=3DBE=4DBE=5

40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

DBE=0

DBE=1

DBE=10

m/z 400.3772 83.66 bins

m/z 400.3767 75.55 bins

-H2

+CH2


Series characterisation



40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

C13C14C15C16C17C18C19C20C21C22C23C24C25C26C27C28C29C30C31C32C33C34C35C36C37C38C39C40

C22 (DBE 0) 77.38 bins 140.88 Å²

C36 175 Å²

C14 100 Å²

C22 (DBE 10) 62.71 bins 122.80 Å²

Carbon number characterisation


Evolution of Travelling Wave Ion Guide Technology

Travelling Wave Collision Cell

Travelling Wave Ion Guide

ScanWave


A Clearer Picture of a complex world


Acknowledgements

Jérémie Ponthus Maíra Fasciotti Priscila Lalli

Mike McCullagh

Eleanor Riches

…and the MS Research Team


Thank You For Your Attention


Analysis of Crude Oil SAR Fraction

IFP uses FTICR-MS to analyse crude oils & their fractions

Collaboration: samples provided by IFP and comparative analyses made with FTICR-MS & SYNAPT HDMS

SAR fraction: — Saturated 44.5%

— Aromatics 36.3%

— Resins 16.4%

Sample: from Nigerian Egina oil field — Vacuum residue

— Resins fraction

— Most polar fraction

— Particularly nitrogen-rich oil

Samples: with thanks to Jérémie Ponthus, IFP


IFP FTICR-MS Analysis

Typical data treatment: Kendrick plots using their in-house “KendrickInside” software

exact Kendrick mass = IUPAC mass x (14/14.01565)

Kendrick mass defect = (nominal Kendrick mass - exact Kendrick mass)

# C : Alkylation D

oubl

e B

ond

Equ

ival

ent

Aro

mat

icity

• E. Kendrick, Anal. Chem., 1963, 35, 2146-2154 • C.A. Hughey, C.L. Hendrickson, R.P. Rodgers, A.G. Marshall, Anal. Chem., 2001, 73, 4676-4681


Waters Analysis of Egina Resins

ASAP ESI



Oil fraction mobilogram: an organised area


288 -20 N1

302 -20 N1

316 -20 N1

288 -6 N1

302 -6 N1

316 -6 N1


Lines in the mobilogram relate to series in a Kendrick plot



Kendrick plot of the Synapt G2 HDMS data

Extraction of a nitrogen containing series (family 1)


20

40

60

80

100

120

140

160

180

100 200 300 400 500 600 700 800 900 100040

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

Excel plot of the DriftScope family 1



40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

DBE=0DBE=1DBE=6DBE=7DBE=8DBE=9DBE=10DBE=11DBE=12DBE=2DBE=3DBE=4DBE=5

40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

DBE=0

DBE=1

DBE=10

m/z 400.3772 83.66 bins

m/z 400.3767 75.55 bins

-H2

+CH2


Excel plot of the DriftScope family 1: series characterisation



40

50

60

70

80

90

100

110

120

130

220 270 320 370 420 470 520 570

C13C14C15C16C17C18C19C20C21C22C23C24C25C26C27C28C29C30C31C32C33C34C35C36C37C38C39C40

C22 (DBE 0) 77.38 bins 140.88 Å²

C36 175 Å²

C14 100 Å²

C22 (DBE 10) 62.71 bins 122.80 Å²

Excel plot of the DriftScope family 1: C number characterisation

Documents

T-WaveTM: From innovative technology to · ©2011 Waters Corporation 2 Our Mission Develop a Keen Understanding of our customers’ needs Work on Purposeful Innovations to solve analytical