Waters Innovation Seminar · Waters Innovation Seminar ... report and saves results to the console...
Transcript of Waters Innovation Seminar · Waters Innovation Seminar ... report and saves results to the console...
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Waters Innovation Waters Innovation Seminar
Mars 2013Mars 2013
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
Agenda
09:00-10:00
— LC/MS product update
10:00-10:30
— Coffee
10:30-11:30
— Lab Efficiency
11:30-12:30
— Lunch
©2011 Waters Corporation 2
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Eftermiddag
12:30-13:20
— Maximize your LC productivity with 2.5µm particles
13:20 Coffee
14:00
— UNIFI – The software that will change the industry standard
14:30-15:30
— Acquity UPC2
15:30-17:00
Aft W k
©2011 Waters Corporation 3
— After Work
Waters LC & MS FamilyWaters LC & MS Family
Per Per RagnebornRagneborn
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
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ACQUITY UPLCACQUITY UPLC
©2011 Waters Corporation 5
Introducing ACQUITY UPLC I-Class:Waters fifth UPLC-version
©2011 Waters Corporation 6
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Why I-Class?
Complex separation challenges require LC systems that are designed to maximize the benefits of sub 2µm particle columns.
Has minimized dispersion to enhance MS and UV performance
Lowest carryover complementing MS sensitivity and extending MS linear dynamic range
©2011 Waters Corporation 7
g y g
The system’s low dispersion and faster cycle-time allows complex separations to be accelerated without compromising chromatographic fidelity
Lower Dispersion I-Class vs Competitor
Competitive UHPLC System: 21μl extra column band spread
72.00
90.00
Average Peak Capacity at 5σ : 42
0.104
0.130
Average Peak Capacity at 5σ: 70
AU
0.00
18.00
36.00
54.00
Minutes0.07 0.14 0.21 0.28 0.35 0.42
Average Peak Capacity at 5σ : 42
©2011 Waters Corporation 8
AU
0.000
0.026
0.052
0.078
Minutes0.00 0.07 0.14 0.21 0.28 0.35 0.42
ACQUITY UPLC I-Class: 5μl extra column band spread
5
AU
0.002
mAU
2.00
ACQUITY UPLC I-Class System Vendor B UHPLC System
Improved Dispersion
0.000
Minutes0.00 2.00 4.00 6.00 8.00
0.00
Minutes0.00 2.00 4.00 6.00 8.00
©2011 Waters Corporation 9
Minutes0.40 0.50 0.60 0.70 0.80
Minutes0.40 0.50 0.60 0.70 0.80
Resolution and sensitivity is improved
Waters Xevo MS FamilyWaters Xevo MS Family
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
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Xevo TQD
The Xevo TQ Family The Xevo TQ Family ––Built on the Same FoundationsBuilt on the Same Foundations
SQ Detector 2
Xevo TQ-S
Xevo TQ MS
©2011 Waters Corporation 11
Frost and Sullivan Product Quality
Leadership Award 2011IBO Best new
product, ASMS 2008
Xevo G2Xevo G2--S (Q) TofS (Q) TofTime Of Flight MSTime Of Flight MS
©2011 Waters Corporation 12
XevoXevo G2G2--S S QTofQTofXevoXevo G2G2--S S TofTof
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Common Xevo features !
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
Enhancing Usability
ZsprayTM
— Robust performance in the face of complex biological samples
— Zspray, a proven geometry (many years experience & 1000’s in use daily).
Designed to be compatible with all your HPLC and UPLC needs
Innovative, Ergonomic
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
, gEngineering— Performance, usability &
serviceability — Tool free maintenance— Novel tool free source exchange— Integrated gases— Plug and play probes
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Optimizing Gas Flow Dynamics
Shape of chamber— Smooth, curved surface
— Reduces turbulence
E ll t b t bilit — Excellent beam stability
Size of chamber— Efficient desolvation
— Excellent linear response
Temperature of chamber— Thermally regulated
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
Tangential exhaust— Gas flows modeled to efficiently
remove non-ionized materials from source chamber
What is IntelliStart?
Continuous background system monitoring Integrated fluidics system for
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
Automated tools to enable quick system start-up
Automated LC/MS system performance checking
g ysample and calibrant delivery
Updated electronics to enable diagnostic functionality
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System Ready to Use
IntelliStart
In- built Fluidics
No extra syringe required
Automated delivery
©2011 Waters Corporation 17
Automated delivery of reference solutions
Automated flow switching
Calibration
Mass calibration
— IntelliStart automatically warns the user when the calibration needs checking
— Calibrant automatically infused from the integrated fluidics system
©2011 Waters Corporation 18
— Automated MS calibration, routinely checked for validity
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Calibration Check
©2011 Waters Corporation 19
LCMS System Check
Automated system performance checking
— Automatically creates a sample batch with multiple injections of a well characterised analyte
— Automatically checks
o Retention Time
o Peak Area
o Peak Width
o Signal / Noise
©2011 Waters Corporation 20
o Peak Area
o Peak Height
— Automatically generates a printed report and saves results to the console log
o Signal / Noise
o Exact Mass
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LC/MS Check Report
©2011 Waters Corporation 21
System Monitoring
Continuously monitors selected instrument and system parametersinstrument and system parameters
— Operate Status
— Gases
— Voltages
— Temperatures
V
— Valid tune
— Valid calibration
— Service date reached
— Fuses
I t l k
©2011 Waters Corporation 22
— Vacuum
— Communications
Where appropriate, checks are configurable
— e.g. set your own time limit for calibration and tune validity
— Interlocks
— Disk space
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SQD 2
©2011 Waters Corporation 23
SQD 2 PerformanceSQD 2 Performance
3000 m/z Acquisition Range
15,000 amu/sec scanning
3ms inter-scan delay
©2011 Waters Corporation 24
20ms pos/neg switching
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©2011 Waters Corporation 25
XevoXevo®® TQD TQD
Xevo TQDKey Features
Tandem quadrupole mass spectrometer
Small footprint :< 350 mm wide— < 350 mm wide
— Same size (width) as the SQD 2
Z-Spray ion sources— Highly robust, change modes in minutes
— ESI and ESCi are standard
— IonSabre II APCI, APPI, ASAP, APGC options
— Integrated fluidics
— Tool free routine maintenance
©2011 Waters Corporation 26
Tool free routine maintenance
2 to 2000 Da mass range
• T-Wave collision cell for Hi-Speed UPLC-MS/MS
Scan rate up to 10,000 Da per second
Polarity switching in 20 ms
3ms Dwell time
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TT--Wave collision cellWave collision cellEnsures UPLC compatibilityEnsures UPLC compatibility
Travelling Wave Ion Transport The effect of MRM acquisition rate on signal intensity
166 data points per second
©2011 Waters Corporation 27
Know More About Your SamplesKnow More About Your SamplesRADAR and PICSRADAR and PICS
RADAR - Unique simultaneous acquisition of MRM and full scan dataacquisition of MRM and full scan data
See what you cannot see in MRM alone
Method Assess risk Is the sample
©2011 Waters Corporation 28
development of matrix effects
pbackground changing?
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Matrix Profile
Simultaneous targeted quantification Simultaneous targeted quantification & full scan monitoring& full scan monitoring
%
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
214.1415.3
363.4
%
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
214.1415.3
363.4
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
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
Matrix ProfileMS full scan negative ESI
Matrix Profile MS full scan positive ESI Plasma matrix
Spectral information
Qual
itat
ive
e
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
m/z100 200 300 400 500 600 700 800 900 1000
0
100.1149.3 279.3
251.1
546.4468.6
600.7
m/z100 200 300 400 500 600 700 800 900 1000
0
100.1149.3 279.3
251.1
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
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
Targeted compoundMRM positive ESI
Fluticasonein plasma
Quan
tita
tive
PIC SCAN:
Information rich dataInformation rich dataPrecursor Ion Confirmation ScanPrecursor Ion Confirmation Scan
MRMAcquisition
MRM data is used as a specific trigger for the acquisition of a product ion spectrum.
©2011 Waters Corporation 30
PIC Scan Acquisition
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Xevo TQ SXevo TQ-S
Imagine sensitivity Previously Beyond Your Reach...
©2011 Waters Corporation 31
Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Xevo TQXevo TQ-S
Larger sampling orifice
©2011 Waters Corporation 32
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Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Off-Axis design
Narrow Ion TunnelConjoined to
Wide Ion Tunnel
©2011 Waters Corporation 33
Maximising signal
Minimising noise
Enhanced sensitivity
Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Reserpine (50fg) Reserpine (50fg) UPLC/MRM, ESI +UPLC/MRM, ESI +
%%
Enhanced sensitivity with StepWaveTM ion optics
>25X increase in peak area>5X increase in signal:noise
ion a
bundan
ce
©2011 Waters Corporation 34
Time1.80 1.90 2.00 2.10 2.20 2.30
0 Time1.80 1.90 2.00 2.10 2.20 2.30
0
Time1.80 1.90 2.00 2.10 2.20 2.30
%
1
Without StepWaveTM
ion optics
Rel
ativ
e
18
100100
Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Prostaglandin (Plasma)Prostaglandin (Plasma)UPLC/MRM, ESI UPLC/MRM, ESI --
%%
Enhanced sensitivity with StepWaveTM ion optics
30X increase in peak area30X increase in signal:noise
ion a
bundan
ce
©2011 Waters Corporation 35
Time0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
0 Time0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
0
Time0.80 1.00 1.20 1.40 1.60 1.80 2.00 2.20 2.40
%
0
Without StepWaveTM
ion opticsRel
ativ
e i
Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
UPLC/MRM of Verapamil(solvent standard) using a properly
conditioned ACQUITY 100100
220 Z t l *
Stepping up to the challengeStepping up to the challenge
1000.83
1000.83
100100
Q
2fg
%
0.83
%
0.83
0.1fg
220 Zeptomoles* on column
6 replicates
RSD<20%
©2011 Waters Corporation 36
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0 Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0 Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
%
0
Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
0 Time0.25 0.50 0.75 1.00 1.25 1.50 1.75 2.00 2.25 2.50 2.75
0
Solvent
Blank
* 10-21 moles
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Class leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Fenuron ESI+ 30 7
Relative Peak Area
Relative S:N
Compound NameIonisation
Mode
Fenuron ESI+ 30 7
Relative Peak Area
Relative S:N
Compound NameIonisation
Mode
Comparing Xevo TQ-S With Xevo TQ
Back to back comparisons made for a number of different UPLC/MRM assays
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 3F t l ESI+ 39 4
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 3F t l ESI+ 39 4
©2011 Waters Corporation 37
assays Formoterol 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
Formoterol 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
Assay RobustnessClass leading UPLC MS/MS performanceClass leading UPLC MS/MS performance
Compound name: OH ProgCorrelation coefficient: r = 0.999149, r̂ 2 = 0.998298Calibration curve: 3386.22 * x + 23.6767Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None
Compound name: OH ProgCorrelation coefficient: r = 0.999149, r̂ 2 = 0.998298Calibration curve: 3386.22 * x + 23.6767Response type: External Std, AreaCurve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None
Verapamil, 10pg/µL spiked into supernatant from 2:1 ACN:Plasma protein
precipitation.
1000 on column injections
RSD of peak areas < 5%
APCI+, Hydroxprogesterone
Triplicate injections of Standards
50fg to 5 000pg on column
Linearity of Response
©2011 Waters Corporation 38
Conc0 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
Res
pons
e
0
500000
1000000
1500000
2000000
2500000
3000000
Conc0 50 100 150 200 250 300 350 400 450 500 550 600 650 700 750 800 850 900 950 1000
Res
pons
e
0
500000
1000000
1500000
2000000
2500000
3000000RSD of peak areas < 5% 50fg to 5,000pg on column
5 orders of magnitude
Correlation coefficient>0.995
Deviation<15%
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Xe oXe o G2G2 S S TofTofXevoXevo G2G2--S S TofTof&&
XX G2G2 S S Qt fQt f
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
XevoXevo G2G2--S S QtofQtof
Introducing Xevo G2-S QTof
©2011 Waters Corporation 40
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ric
Fie
ld
StepWave Ion OpticsE
lect
r
Diffuse Ion
Cloud
Maximising signal
©2011 Waters Corporation 41
Maximising robustness
Xevo G2Xevo G2--S S QTofQTofSensitivity Sensitivity EnhancementsEnhancements
G2G2
G2-S
7x area counts10x signal intensity
17x S/N (RMS)
©2011 Waters Corporation 42
5 ng/mL Propranolol in Plasma with Ballistic
Gradient
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QuanTofQuanTof Technology, for Exact Mass Technology, for Exact Mass Quantitative PerformanceQuantitative Performance
Compound name: AlprazolamCorrelation coefficient: r = 0.999587, r^2 = 0.999174Calibration curve: 1.18828 * x + -0.0262545Response type: Internal Std ( Ref 1 ), Area * ( IS Conc. / IS Area )Curve type: Linear, Origin: Exclude, Weighting: 1/x, Axis trans: None
onse
300
400
500
600
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
Alprazolam spiked into plasma: Calibration curve showing 4 orders of linear dynamic range for quantitative studies.
Standard concentrations are between 0.05 and 500 ng/mL
ng/mL0 50 100 150 200 250 300 350 400 450 500
Res
p
0
100
200
300
InIn--Spectrum Dynamic Range (ESI)Spectrum Dynamic Range (ESI)
Results and Discussion
Introduce the analytical problem i.e. detection, quantification,
identification and characterization of small molecule form
1.5 ppm<0.1ppm
2 53 E6complex rat bile sample.
Run through the importance of high resolution and wide spectral
dynamic range for the analysis of complex mixtures
Introduce the importance of mass measurement accuracy and
stability.
Introduce the need for comprehensive MS/MS - MSE
OO
O
ON
N
Verapamil
O
O N
NNN
Caffeine
2.53 E6
>104
©2011 Waters Corporation 44
1.70 E2
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MSMSE E
Comprehensive
—Exact mass precursor & product ions
Data-independent – NOT Data dependent like DDA
—No selection of precursors
©2011 Waters Corporation 45
Fast
—UPLC compatible
All of the data, all of the time
QTofQTof MSMSEE mode acquisitionmode acquisition
©2011 Waters Corporation 46
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Resolution— 32.500 FWHM
No compromises !No compromises !
Mass accuracy— Better than 1 ppm RMS both in MS and MS/MS
— No sacrificing in sensitivity
32.500 FWHM— Full resolution maintained without loosing dynamic
range, mass accuracy or sensitivity
In spectrum dynamic range
©2009 Waters Corporation | COMPANY CONFIDENTIAL©2011 Waters Corporation
p y g— 5 orders of magnitude
— Without sacrificing resolution
Scan rates— 30 scans/sec
— No differences between MS or MS/MS
The next step in the evolution of
©2011 Waters Corporation 48
pthe SYNAPT G2 family
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SYNAPT G2-S…transforming your capacity for discovery, understanding and success
©2011 Waters Corporation 49
Integration of Integration of StepwaveStepwave
©2011 Waters Corporation 50
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Identification …limits of detection
SYNAPT G2-S vs SYNAPT G2
SYNAPT G2-S
©2011 Waters Corporation 51
SYNAPT G2
Quantification…LOQ, linearity and reproducibility
©2011 Waters Corporation 52
MS Mode s/n 1pg on column
On column LOQ (fg)
SYNAPT G2-S Resolution MSE >600 25 fg
SYNAPT G2 Resolution MSE >100 250 fg
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Increasing information content (HDMSIncreasing information content (HDMSEE))……High Definition UPLC/MSHigh Definition UPLC/MSEE analysisanalysis
m/z m/z
Q1 RF only
©2011 Waters Corporation 53
Drift time
Precursor ions separated by IMS
m/z
Precursor ionsDrift time
/
Precursor and products share same drift time
Separation of isomers at same retention time and exact mass.
* *
164.55Å 174.1Å
©2011 Waters Corporation 54
HDMS data shows two isomers present at R.T
3.25 mins
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UPLC/HDMSE
©2011 Waters Corporation 55
Ion mobility
Universal Source platform
Compatibility across the boardCompatibility across the board
ESIAPCIESCi™
APPIAPCI
APGC TRIZAIC UPLC®
nanoFlow™ESI
ASAP
©2011 Waters Corporation 56
Also compatible with 3rd party source designs:
Phytronix LDTD IonSense DART Prosolia DESI Advion NanoMate
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©2011 Waters Corporation 57