development of the worldthe world s’smost most lc/ms ... · 5 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2...
TRANSCRIPT
![Page 1: Development of the Worldthe World s’sMost Most LC/MS ... · 5 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4 Acquisition Time (min)Time (min) ... Photo of new 6490 taken from 1 meter](https://reader031.vdocuments.us/reader031/viewer/2022011802/5b16065d7f8b9a961e8c95f1/html5/thumbnails/1.jpg)
Development ofthe World’s Mostthe World s Most
SensitiveTriple Quad MS
LC/MS Divisionand Agilent LabsAgilent TechnologiesTriple Quad MS Santa Clara, CA
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New 6490 - World’s Most Sensitive LC/QQQ
iFunnel TechnologyAgilent Jet StreamHexabore capillaryDual-stage ion funnel
Collision Cell – hexapoleTapered structure for increased ion acceptanceReduction of background noise
Improved Quad Drive ElectronicsImproved Quad Drive ElectronicsHigher RF power capabilityQuad drive frequency increased to 1.4 MHz
2
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NEW IF UNNE L T ECHNOLOGYiFunnel Technology captures 10X more ions
Agilent Jet Stream Hexabore Capillary Dual Ion Funnel
• Thermal confinement of ESI plume
• Efficient desolvation to
• 6 capillary inlets• Samples 6X more ion rich
gas from the source
• Removes the gas but captures the ions
• Removes neutral noisecreate gas phase ions
• Creates an ion rich zone
gas from the source• Captures the majority of
the gas from the source region
• Removes neutral noise• Extends turbo pump life
region
3
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Agilent Jet Stream Ion Generation
Gas Dynamics View
N b li iNebulizing gas
S h d h h
Enhanced efficiency nebulizer
Nozzle voltage
Super-heated sheath gas
The super-heated sheath gas
g
Heated drying gas
e supe eated s eat gascollimates the nebulizer spray and creates a dramatically “brighter source” Resistive sampling
capillaryPatent Pending
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Agilent’s Hexabore Atmospheric Sampling…. The Most Sensitive…. By Far.
6 Bore Capillary
y
• Nearly 6 x the amount of atmospheric gas sampledp g pcompared to a single capillary systemANDAND
• 5 - 10x more ions into the high vacuum regiong g
5
But how do we handle all the extra gas molecules?
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Need for Greater Ion Sampling Capacity
• Many ions formed in the collimated Agilent Jet Stream plume are still not captured by the MS system
• Ions sampled by a single capillary with a 600 µm i.d.
• Maximum ion generation occurs across a 3-5 mm horizontali t th t f th lregion at the center of the plume.
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Hexabore Sampling Capillary
• Hexabore sampling capillary increases ion sampling efficiency about 6Xion sampling efficiency about 6X compared to single bore
Ion sampling gains are mostIon sampling gains are most dramatic with the combination of both Agilent Jet Stream technology with the hexabore sampling p gcapillary
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Hi h L
Agilent’s Two Stage Ion Funnel Can Handle the Gas Load
High Pressure Stage 1
Low Pressure Stage 2
ExistingRF IonGuide
St 1Stage 2
Stage 18-12 Torr
1-3 Torr
Stage 1 offset breaks up the high pressure gas exiting the Hexabore Capillary
8
Stage 1 offset breaks up the high pressure gas exiting the Hexabore Capillary
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How Does an Ion Funnel Work?RF DriveThe RF Voltage
focuses the ions to the center.
UnfocusedIons and GasEnter
FocusedIons Out
The DC Voltage accelerates theaccelerates the ions to the exit. DC Drive
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How is an Ion Funnel Constructed?Gold Plated
Previously, with many metal plates…. Rim
l i i l i i l d d… resulting in a large capacitive load and relatively large RF power electronics.
The 6490 design uses printed circuit board technology with only a small conductive rim resulting in a reduced capacitance load. This makes +/- ion switching possibleThis makes +/ ion switching possible.
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Improved Ion Sampling Efficiencies with iFunnel
Agilent Jet Stream positive ion
Agilent Jet Stream negative ion
14x11x
10x7x5x
7x
7x7x 6490
10x
5x
302 10341222
922622
322
118 6460
200 400 600 800 1000 1200 1400
113
200 400 600 800 1000 1200 1400m/zm/z
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6490 with iFunnel vs 6460 in positive ion mode
1 pg Reserpine (7X)
103
1 pg Alprazolam (6X)
3x103
3
6490
x103
6490
2
16460
A i iti Ti ( i )0 0.1 0.2 0.3 0.4 0.5 0.6
Acquisition Time (min)0 0.2 0.4 0.6 0.8 1.0 1.2
6460
Acquisition Time (min)
12
Acquisition Time (min)
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New 6490 Increases Signal but also Reduces Noise
6X gain in signal shown in previous slide translates into 10X gain in signal-to-noise for alprazolam relative to the 6460 as shown below
6460S/N = 1,157:1 where noise = 1xRMS
6490S/N = 11,640:1 where noise = 1xRMSwhere noise = 1xRMS where noise 1xRMS
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6490 with iFunnel vs 6460 in negative ion mode
1 pg Chloramphenicol (8X) 1 pg Acid red 4 (9X)
3x103
3x103
T it
Relative Increase in
36490
3
0.91
1.1 6490
Tritonsignal of Triton vs. 6460
6 x
2
0.60.70.8
6460 BB1: CAT CC
1
0.30.40.5
CAT CConly
00 5 1 1 5 2
6460
00.10.2
0 2 0 4 0 6 0 8 1
6460
0.5 1 1.5 2Acquisition Time (min)
0.2 0.4 0.6 0.8 1Acquisition Time (min)
14
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New iFunnel Technology RevolutionizesAtmospheric Samplingp p g
The combined result of all three elements of iFunnel Technology is aThe combined result of all three elements of iFunnel Technology is a dramatic gain in sensitivity.
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Novel Curved and Tapered Collision Cell DesignEfficient Ion Transmission over wide mass rangeReduces transmission of ionizer generated noiseReduces transmission of ionizer generated noise
RF & DCVoltages Set
Confinement and Axial Acceleration
Larger Smaller (3.5mm)
(5 mm)
EntranceAperture
Accepts more
( )
ExitApertureFocusesI i t Accepts more
IonsIons intoMS2
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New 6490 LC Triple with iFunnel Technology “Unrivaled Quantitative Excellence”Unrivaled Quantitative Excellence
1290 Infinity LC/6490 Triple : Clearly Better Together
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Agilent Triple Quads: 2006 – 2010Relentless increase in signal-to-noise specsRelentless increase in signal to noise specs
6490
8000
9000
10000
rpin
e
4000
5000
6000
7000
1 pg
rese
r
1000
2000
3000
4000
64106430
6460
SNR
1
0
2006 2007 2008 2009 2010
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New 6490 Triple installation Spec: S/N > 10,000:1 so the 6490 should be able to detect 1 fg ?so the 6490 should be able to detect 1 fg ?
1 pg reserpine on-columnX103
1 pg reserpine on column S/N > 22,605:1
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6490 Triple - Attogram Sensitivity for Reserpine1 f f i i j t d l1 fg of reserpine injected on-column
S/N scales as expected
NO gaming the noise spec
Acquisition Time [min]
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6490 Achieves Zeptomolar Detection Limits
200 zeptomoles
+ MRM (455.3 -> 164.9)1
x10
p
verapamilInjected on-column
4.5
4.4
4 2
4.3
Acquisition Time (min)0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
4.2
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Unprecedented Six logs of Linearity with 6490 100 Attogram to 100 Picogram arapmil on col mn
Y = 65283 x – 0.390412R2 = 0 99761000000
10000000
100 Attogram to 100 Picogram varapmil on-column
R = 0.9976
nse 100000
1000000
ea R
espo
n
1000
10000
Are
10
100
Femtograms
10.1 1 10 100 1000 10000 100000
e tog a s
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50 pg/mL Fluticasone in plasma with 6460
25 Femtogram on-columnArea RSD = 23.7% (n=6)Mean accuracy = 104
Fluticasone propionate
*1.534 min.1x10
5.2
5.3
OHCH3
OS
F
O
O
CH3
Fluticasone propionate
4.8
4.9
5
5.1
O
CH3
F
F H
CH3O
4.4
4.5
4.6
4.7
MRM unit / unit resolution (0 7 amu / 0 7 amu)
Acquisition Time (min)0.7 0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4
4.2
4.3 (0.7 amu / 0.7 amu)
Acquisition Time (min)
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5 pg/mL Fluticasone in plasma with new 64902.5 Femtogram on-column
Area RSD = 8.7% (n=5)Mean accuracy = 104
Fl ti i t1x10
5.2
5.3
1.765 min.
CH
OHCH3
OS
F
O
CHO
CH3 MRM enhanced / unit resolution (0 4 amu / 0 7 amu)
Fluticasone propionate 1x10
5 1
5.2
5.3
4 7
4.8
4.9
5
5.1
O
CH3
F
F H
CH3(0.4 amu / 0.7 amu)
4 7
4.8
4.9
5
5.1
4 3
4.4
4.5
4.6
4.7
4 3
4.4
4.5
4.6
4.7
Acquisition Time [min]
0.8 0.9 1 1.1 1.2 1.3 1.4 1.5 1.6 1.7 1.8 1.9 2 2.1 2.2 2.3 2.4 2.5
4.2
4.3
4.2
4.3
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q [ ]
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The Value of Enhanced Resolution on 6490Fluticasone propionate (inhalant) in human plasma 2 5 fg on-column)
1
Fluticasone propionate (inhalant) in human plasma 2.5 fg on column)
1
Enhanced Resolution gives increased selectivity for analytes in complex matrices1
x10
7Q1 Unit (0.7 amu)Q2 Unit (0.7 amu)
1x10
7
analytes in complex matrices
6Q1 Enhanced (0.4 amu)Q2 Unit (0.7 amu)6
55
0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4
Acquisition Time (min)0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 1.8 2 2.2 2.4
Acquisition Time (min)
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Ultra Trace Levels of Pesticides in Potable Water
Atraton S/N =: 819
Triazine herbicides analyzed at 50 parts per quadrillion
6490 T i l Q dAtraton S/N : 819
Simazine S/N = 18
6490 Triple Quad with on-line enrichment
Simetryn S/N = 9
Secbumeton S/N = 51
Prometon S/N = 40
Atrazine S/N = 28
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Peptide MRM on 6460 vs new 6490 Triple Quad
LVNEVTEFAK
1x10
4.5
1 femtomole on 6460
LVNEVTEFAK575.5 937.5
4.3
4.4
4.2
110 1x10
4.5
100 attomole on 6490
4.3
4.4
4.2
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Triggered MRM – What Problem Does tMRM Solve?
T t d Q t C d Id tifi ti
Data dependent product ion scans leaves gaps in MRMi f d th ti i t
Targeted Quant + Compound Identification
Gap in MRM trace, numerically filled in
• mix of raw and synthetic points• higher RSDs and higher LOQs
numerically filled in.Typically > 100 msec
DDADDA –scan switch to PIS
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tMRM for up to 8 Confirmatory Ion Transitions- when primary compound MRM triggers confirmatory MRMswhen primary compound MRM triggers confirmatory MRMs
Plot counts versus m/z for product ions
Faster compound ID at higher sensitivity with tMRM
Plot counts versus m/z for product ions “Composite MS/MS spectra for compound b”
Product %MRMIon Scan Sensitivity
ts
yTime
QTRAP 5500 ~ 100 msec ~ 10%
Vantage ~ 150 msec ~ 2%
coun
t Vantage ~ 150 msec ~ 2%Xevo ~ 150 msec ~ 2%
6490 ~ 30 msec 100%
100 200 300
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The 6490 has Zeptomole Capabilitywhat is a zeptomole?p
Photo of new 6490 taken from 1 meter distanceHow far away is 1021 meters?
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The Milky Way Galaxy as seen from Hubble1021 meters from the earth is at the edge of the known universe10 meters from the earth is at the edge of the known universe
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The New 6490 Triple Quad System
New era in sensitivity – Zeptomole
New era in linearity – Six decades of linearityNew era in linearity Six decades of linearity
New iFunnel Technology
Improving compound identity tMRMsImproving compound identity - tMRMs
“Delivers Unprecedented Quantitative Excellence”