Transcript LCMS Overview Jan 2013

Agilent LC/MS
Overview
Nathan Miller
LC/MS Product Specialist
Agilent’s > 40 year heritage in mass spectrometry
1971, 5930A
ICP-MS 1994 - 2011
GC/MS 1971 - 2011 2008, 7000A
2005, 5975A
1994, 4500 Series
2011
7200A
1976,
5992A
2012 8800 QQQ
2000, 7500 Series
LC/MS 1980 - 2011
Direct Liquid Injection
1980, 5985 SQ
ESI on 5989
MS Engine
LC/QQQ
2003
1994, 2000
2006
Thermospray, 5987
LC/MSD TOF
Particle Beam, 5988
LC/MSD, LC/MSD Ion Trap
LC/QTOF
Over 37,000 GC/MS, 4,500 ICP-MS and 6,500 LC/MS systems through June 2012
Agilent (and previously, as HP) has sold more quadrupole MS systems than any other mfr.
Agilent High Performance Infinity Series LC:
Best in class “Front End” systems for MS
Preparative LC
Standard LC
Ultra High Pressure
LC
Capillary &
Nano LC
High Sensitivity
HPLC-Chip/MS
The industry’s most comprehensive LC portfolio to configure an
integrated LC/MS system
MassHunter Workstation
One software for all Agilent Mass Specs
Minimize the learning and optimize the use of software in your
lab across different mass spec instrument platforms
• Control of Agilent 6000 Series QQQ, TOF and Q-TOF
• Control of most 1260 and 1290 HPLC modules, 7100 CE
• Control for GC QQQ and Q-TOF
• Data Processing for QQQ, TOF, Q-TOF, SQ*) and GC Ion Trap*)
• Control & data processing for ICP/MS
LC/MS
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GC/MS
ICP-MS
conversion to
*) after
MassHunter format
LCMS Overview Jan 2013
7/6/2015
Agilent 6000 Series LC/MS portfolio
High Resolution
6200 series TOF
6500 series Q-TOF
Quantitative
6100 series Quad Low Resolution 6400 series QQQ
Tandem Stage
Single Stage
Qualitative
Single Quad LC/MS – Proven Solutions
Single Quad
Control Software
ChemStation
6120B – 2,000 m/z
6130B – 3,000 m/z
Easy Access
G2725AA
Single Quad
Data Analysis
Software
MassHunter
Data Translator
Analytical Studio
Reviewer, G3772AA
6150B – 1,400 m/z
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LCMS Overview Jan 2013
7/6/2015
Single Quad Supports Wide Range of Ion Sources
seven Agilent ion sources and third party sources
MultiMode
HPLC Chip
Plus Direct Sampling
Sources from Ion Sense:
APCI
APPI
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nanoESI
ESI
AJS
LCMS Overview Jan 2013
7/6/2015
Agilent Jet Stream
Gas Dynamics View
Nebulizing gas
Enhanced efficiency nebulizer
Super-heated sheath gas
Nozzle voltage
Heated drying gas
The super-heated sheath gas collimates
the nebulizer spray and creates a
dramatically “brighter source”
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Resistive sampling
capillary
LCMS Overview Jan 2013
7/6/2015
iFunnel Technology
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LCMS Overview Jan 2013
7/6/2015
iFunnel Technology Revolutionizes Ion Sampling
Agilent Jet Stream
Hexabore Capillary
• Thermal confinement of •
ESI plume
•
• Efficient desolvation to
create gas phase ions
6 capillary inlets
Samples ~10 times
more ion rich gas from the
source
Dual Ion Funnel
• Removes the gas but
captures the ions
• Removes neutral noise
• Extends turbo pump life
• Creates an ion rich zone • Captures the majority of
the gas from the source
region
Captures up to 10 x more ions
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LCMS Overview Jan 2013
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6230B Accurate Mass TOF LC/MS
True Hi-Def TOF Technology:
NO PERFORMANCE COMPROMISE!
•
Unmatched Mass Accuracy
< 1 ppm MS
•
>22,000 Resolution
•
105 Dynamic Range
•
Up to 30 spectra/second
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LCMS Overview Jan 2013
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Formulas for TOF performance
922.0092
Resolving power =
(m/z)/w1/2
W1/2 = 0.022
1000/0.025 = 40,000
923.0121
922/0.022 = 41,909
921.5 922.0 922.5 923.0 923.5 924.0 924.5
m/z, amu
+TOF MS: 5.474 to 5.547 min from steroid2TOFap42.wiff Agilent, subtracted (4.696 to ...
1.10e5
1.00e5
9.00e4
In te n s ity , c o u n ts
 actual- measured
6
Mass accuracy 
 10
actual


289.2159
1.16e5
Mass accuracy ( rel. mass error)
Max. 1.2e5 counts.
Epitestosteron
e
8.00e4
7.00e4
Resolving
power: 6890
6.00e4
(1000.000-1000.001)/1000)x106=1 ppm
5.00e4
4.00e4
271.2051
3.00e4
2.00e4
Therefore: 1 ppm @ 1000 m/z is 0.001Da
(1 mDa)
1.00e4
0.00
120
actual
289.2162
meas
289.2159
-1.06 ppm
140
160
180
200
220
269.1893
287.1997
240
260
m/z, amu
280
300
320
340
360
380
400
Rapid Analysis of Pharmaceutical Compounds
TOF - MS with Peak Capacity of 40 Cycle in 39 seconds
7.0e5
6.5e5
6.0e5
5.5e5
Metoprolol
Verapamil
0.36s
0.42s
5.0e5
4.5e5
Intensity, cps
4.0e5
Atenolol
3.5e5
Beclomethasonedipropionate
3.0e5
0.34s
2.5e5
0.36s
Primidone
2.0e5
0.36s
1.5e5
1.0e5
5.0e4
0.0
0.05
0.10
0.15
0.20
0.25
0.30
0.35
Time, min
Page
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14
0.40
0.45
0.50
0.55
0.60
6000 Overview Diploma Training 2006
Agilent Restricted
LCMS Overview
Jan 2013
7/6/2015
A big leap forward in 2006
6100 Series SQ
Collision cell incorporates axial
acceleration for high speed MS/MS
analysis
Quad Mass Filter (Q1)
Octopole 1
Lens 1 and 2
Rough
Pump
Turbo
1
Turbo
1
10KV
Detector
6500 Series Q-TOF
6400 Series QQQ
Turbo
1
Quad Mass Filter (Q1)
Quad Mass Filter (Q3)
Octopole 1
Lens 1 and 2
Rough
Over 15 Years in LC/MS Pump
atmospheric sampling
and patented orthogonal
geometry - Result in
industry leading sensitivity
and robustness
Turbo
1
10KV
Detector
Collision
Cell
Turbo
1
Turbo
1
Quad Mass Filter (Q1)
Octopole 2
Octopole 1
Lens 1 and 2
Rough
Pump
Turbo
1
Turbo
1
Collision Cell
Turbo
1
DC Quad
Ion Pulser
Turbo
2
Performance
Agilent 6400 Triple Quadrupole Product Line
A New Agilent LC
Triple Quad
Launched
Every Year
For last
4 years
6490
Triple
Quad
6460 6460
TripleTriple
QuadQuad
6430
Triple
Quad
6420
Triple
Quad
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LCMS Overview Jan 2013
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What is MRM? (Multiple Reaction Monitoring)
Ions
Spectrum with
background
ions (from ESI)
Q1 lets only
target ion 210
pass through
210
Collision cell
breaks ion 210
apart
210
222
268
165
170
210
250
158
280
290
191 210
190 210
150
170
190
210
Q3 monitors only
characteristic
fragments 158
and 191 from ion
210 for quant and
qual.
158
160
191
190
no chemical background
For Research Use Only.
Single quads vs. Triple quads in complex matrix
SIM m/z 306
m/z 306 → m/z 261
MS-MS
Triple quads enjoy the benefit of filtering out
the noise as observed in SIM
Triggered MRM (tMRM) Acquisition:
Simultaneous Quantitation and Confirmation
Triple Quad MRM
Acquisition
Data
Dependent
MRM
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Dynamic
MRM
Triggered
MRM
LCMS Overview Jan 2013
7/6/2015
Triggered MRM (tMRM)
Fast, Sensitive Compound Confirmation to Prevent False Positives
Triggered cycle (above threshold)
Additional MRMs
used
for compound
confirmation
Threshold
Primary cycle (below threshold)
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LCMS Overview Jan 2013
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tMRM Product Ion Spectrum
A Re-Constructed Product Ion Spectrum that is Library Searchable
x103
8
7
6
5
4
3
2
1
0
x103
8
7
6
5
4
3
2
1
0
x103
8
7
6
5
4
3
2
1
0
334 > 76
76.0
80
334 > 132
132.1
100
140
91.1
80
100
334 > 145
x103
8
7
6
5
4
3
2
1
0
100
334 > 105
x103
8
7
6
5
4
3
2
1
0
145.0
140
180
105.1
80
100
120
147.0
100
140
145.0
76.0
91.1 105.1
x103
8
7
6
5
4
3
2
1
0
334 > 147
x103
8
7
6
5
4
3
2
1
0
180
334 > 117
x103
8
7
6
5
4
3
2
1
0
117.0
80
x103
8
7
6
5
4
3
2
1
0
100
120
334 > 119
119.0
80
100
334 > 171
171.1
80
100
120
140
160
120
140
180
Tebufenpyrad
132.1
171.1
100
120
tMRM Product Ion
Spectrum
117.0
80
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334 > 91
x103
8
7
6
5
4
3
2
1
0
160
180
334.2
200
220
240
260
280
300
320
340
LCMS Overview Jan 2013
7/6/2015
6500 QTOF Product Line
Excellent Value and Performance
6530 QTOF LC/MS
 Agilent JetStream for greatest sensitivity
 Robust, easy to use
 Wide dynamic range for qual/quan applications
6540 UHD QTOF LC/MS
 Sub 1-ppm mass accuracy for highest confidence
compound identification
 Versatile system with highest resolution
 Fast scan speed to match UHPLC separations
6550 iFunnel QTOF LC/MS




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Agilent iFunnel for highest sensitivity
Perfect for most demanding applications
Wide dynamic range for qual/quan applications
Fastest scan speed to match UHPLC separations
LCMS Overview Jan 2013
7/6/2015
New 6550 iFunnel QTOF
10X Sensitivity Gain Enables Applications
Sensitivity
• Dramatically improved quantitative capabilities
• New Qual/Quan Workflows
• Superior metabolite and protein detection
• Non-targeted compound screening
Comprehensive Performance Enhancements
• Mass Resolution >40,000
6550 iFunnel Q-TOF
LC/MS System
• 50 spectra /sec MS and 33 spectra/sec MS/MS
• 5 orders of linear dynamic range
• <1 ppm MS mass accuracy; <2 ppm MS/MS
• Unrivalled sensitivity
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ASMS
2010
LCMS Overview
Jan 2013
7/6/2015
Low Femtogram Detection Level
MS
EIC
Page 24
25 fg Reserpine
Resolution
~ 31,500
S/N = 49 (RMS)
Mass error
-0.1 ppm
LCMS Overview Jan 2013
7/6/2015
G3212A - GC APCI Interface
•Increases MS
flexibility by enabling
both GC and LC
compatibility
•Analysis of wider
range of compounds
•Proven QTOF LC/MS
platform
•Proven 7890 GC
performance
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LCMS Overview Jan 2013
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GC Transfer line / APCI Source Design
Corona needle assembly
Capillary MS inlet
Bayonet lock coupling
Transfer line heat shield
Heated transfer line
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LCMS Overview Jan 2013
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Simplify Your Startup
Method
Databases
and
Libraries
LC
Column
Page 27
Standards
LC/MS
Application
Kits
On Site
Training
LCMS Overview Jan 2013
7/6/2015
RapidFire® Mass Spectrometry
Fast autosampler & SPE system
• Replaces LC in LC-MS
• Integrated, automated, micro-scale solid-phase
extraction
• Integrates with standard ESI MS instruments
• Compatible with most biological matrices
• cycle time: 5–10s/sample
The RapidFire System:
• Fits existing workflows
• Low operational costs
• Improved data quality
• Strong correlation with traditional methods
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LCMS Overview Jan 2013
7/6/2015
RapidFire™ Mass Spectrometry (RF-MS)
Microfluidic Sample
Clean-up System
Agilent 6410
QqQ Mass Spectrometer
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96 or 384 well
Plate Handler
Monitor to track
MSMS data
Monitor
to track
RapidFire System
LCMS Overview Jan 2013
7/6/2015
THANK YOU!
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LCMS Overview Jan 2013
7/6/2015