Development of
the World’s
World s Most
Sensitive
Triple Quad MS
LC/MS Division
and Agilent Labs
Agilent Technologies
Santa Clara, CA
New 6490 - World’s Most Sensitive LC/QQQ
iFunnel Technology
¾ Agilent Jet Stream
¾ Hexabore capillary
¾ Dual-stage ion funnel
Collision Cell – hexapole
¾ Tapered structure for increased ion acceptance
¾ Reduction of background noise
Improved Quad Drive Electronics
¾ Higher RF power capability
¾ Quad drive frequency increased to 1.4 MHz
2
NiFunnel
E W I F UTechnology
N N E L T E C Hcaptures
N O L O G Y 10X more ions
Agilent Jet Stream
• Thermal confinement
of ESI plume
• Efficient desolvation to
create gas phase ions
Hexabore Capillary
Dual Ion Funnel
• 6 capillary inlets
• Removes the gas but
captures the ions
• Samples 6X more ion rich
gas from the source
• Captures the majority of
• Creates an ion rich zone
the gas from the source
region
3
• Removes neutral noise
• Extends turbo pump life
Agilent Jet Stream Ion Generation
Gas Dynamics View
Enhanced efficiency nebulizer
N b li i gas
Nebulizing
S
Super-heated
h
d sheath
h h gas
Nozzle voltage
g
Heated drying gas
The
e supe
super-heated
eated s
sheath
eat gas
collimates the nebulizer spray and
creates a dramatically “brighter source”
Patent Pending
Resistive sampling
capillary
Agilent’s Hexabore Atmospheric Sampling….
The Most Sensitive…. By
y Far.
6 Bore Capillary
• Nearly 6 x the amount of
p
gas
g sampled
p
atmospheric
compared to a single
capillary system
AND
• 5 - 10x more ions into
g vacuum region
g
the high
But how do we handle all the extra gas molecules?
5
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 horizontal
region
i att th
the center
t off the
th plume.
l
Hexabore Sampling Capillary
• Hexabore sampling capillary increases
ion sampling efficiency about 6X
compared to single bore
Ion sampling gains are most
dramatic with the combination of
both Agilent Jet Stream technology
with the hexabore sampling
p g
capillary
Agilent’s Two Stage Ion Funnel Can Handle the Gas Load
High Hi
h
Pressure Stage 1
LLow Pressure Stage 2
Existing
RF Ion
Guide
Stage 1
St
8-12 Torr
Stage 2
1-3 Torr
Stage 1 offset breaks up the high pressure gas exiting the Hexabore Capillary
8
How Does an Ion Funnel Work?
The RF Voltage
focuses the ions
to the center.
RF Drive
Unfocused
Ions and Gas
Enter
The DC Voltage
accelerates the
ions to the exit.
Focused
Ions Out
DC Drive
How is an Ion Funnel Constructed?
Previously, with many metal plates….
… resulting
l i iin a llarge capacitive
i i lload
d and
d
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 possible.
possible
Gold Plated Rim
Improved Ion Sampling Efficiencies with iFunnel
Agilent Jet Stream
positive ion
Agilent Jet Stream
negative ion
11x
7x
7x
7x
6490
5x
14x
10x
7x
5x
10x
622
118
6460
922
322
302
1034
1222
113
200
400
600 800 1000 1200 1400
m/z
200 400 600 800 1000 1200 1400
m/z
6490 with iFunnel vs 6460 in positive ion mode
1 pg Reserpine (7X)
1 pg Alprazolam (6X)
x10
103
x103
6490
6490
3
2
1
6460
6460
0
0.1
0.2
0.3
0.4
0.5
0.6
A
Acquisition
i iti Time
Ti
(min)
( i )
12
0
0.2
0.4
0.6
0.8
1.0
1.2
Acquisition Time (min)
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
6460
S/N = 1,157:1
6490
S/N = 11,640:1
where noise = 1xRMS
where noise = 1xRMS
6490 with iFunnel vs 6460 in negative ion mode
1 pg Chloramphenicol (8X)
x103
x103
3
2
6490
Relative
Increase in
signal of
Triton vs. 6460
T it
Triton
6x
1
6460
0
14
1 pg Acid red 4 (9X)
BB1:
CAT CC
only
6460
0.5
0
5
1
15
1.5
2
Acquisition Time (min)
3
1.1
1
0.9
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
6490
6460
0.2
0
2 0.4
0 4 0.6
0 6 0.8
08 1
Acquisition Time (min)
New iFunnel Technology Revolutionizes
Atmospheric
p
Sampling
p g
The combined result of all three elements of iFunnel Technology is a
dramatic gain in sensitivity.
Novel Curved and Tapered Collision Cell Design
Efficient Ion Transmission over wide mass range
Reduces transmission of ionizer generated noise
RF & DC
Voltages Set
Confinement and
Axial Acceleration
Smaller ((3.5mm) )
Exit
Aperture
Focuses
I
Ions into
i t
MS2
Larger (5 mm) Entrance
Aperture
Accepts more
Accepts more Ions
New 6490 LC Triple with iFunnel Technology
“Unrivaled
Unrivaled Quantitative Excellence”
Excellence
1290 Infinity LC/6490 Triple
:
Clearly Better Together
Agilent Triple Quads: 2006 – 2010
Relentless increase in signal-to-noise
signal to noise specs
6490
SNR 1 pg reserrpine
10000
9000
8000
7000
6000
5000
4000
6460
3000
2000
1000
6430
6410
0
2006
2007
2008
2009
2010
New 6490 Triple installation Spec: S/N > 10,000:1
so the 6490 should be able to detect 1 fg ?
X103
1 pg reserpine on-column
on column
S/N > 22,605:1
6490 Triple - Attogram Sensitivity for Reserpine
1 ffg off reserpine
i iinjected
j t d on-column
l
S/N scales as expected
NO gaming the noise spec
Acquisition Time [min]
6490 Achieves Zeptomolar Detection Limits
+ MRM (455.3 -> 164.9)
1
x10
200 zeptomoles
p
4.5
verapamil
Injected on-column
4.4
4.3
42
4.2
0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
Acquisition Time (min)
Unprecedented Six logs of Linearity with 6490
arapmil on
col mn
100 Attogram to 100 Picogram varapmil
on-column
10000000
Y = 65283 x – 0.390412
R2 = 0.9976
0 9976
Are
ea Respon
nse
1000000
100000
10000
1000
100
10
1
0.1
1
10
100
Femtograms
e tog a s
1000
10000
100000
50 pg/mL Fluticasone in plasma with 6460
25 Femtogram on-column
Area RSD = 23.7% (n=6)
Mean accuracy = 104
Fluticasone propionate
Fluticasone propionate F
x10 1
*1.534 min.
5.3
5.2
HO
O
CH3
O
5
F
CH3
O
CH3
CH3
5.1
4.9
S
H
O
F
4.8
4.7
4.6
4.5
MRM unit / unit resolution
(0.7
0.7
(0 7 amu / 0
7 amu)
4.4
4.3
4.2
0.7
0.8
0.9
1
1.1
1.2
1.3
1.4
1.5
1.6
1.7
1.8
Acquisition Time (min)
1.9
2
2.1
2.2
2.3
2.4
5 pg/mL Fluticasone in plasma with new 6490
2.5 Femtogram on-column
Area RSD = 8.7% (n=5)
Mean accuracy = 104
Fl
ti
i
t
Fluticasone propionate x10 1
x10 1
5.3
5.3
5.2
5.2
51
5.1
5.1
5
5
4.9
4.9
4.8
4.8
4.7
47
4.7
47
4.6
4.6
4.5
4.5
4.4
4.4
43
4.3
43
4.3
4.2
4.2
1.765 min.
F
HO
S
O
CH3
O
O
CH3
CH3
F
CH3
MRM enhanced / unit resolution
(0 4 amu / 0.7
(0.4
0 7 amu)
H
O
F
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
Acquisition
q
Time [[min]]
Page 24
The Value of Enhanced Resolution on 6490
Fluticasone propionate (inhalant) in human plasma 2
5 fg on-column)
2.5
on column)
1
x10
Enhanced Resolution gives increased selectivity for
analytes in complex matrices
Q1 Unit (0.7 amu)
Q2 Unit (0.7 amu)
7
Q1 Enhanced (0.4 amu)
Q2 Unit (0.7 amu)
6
5
0.2
0.4
0.6
0.8
1
1.2
1.4
1.6
1.8
Acquisition Time (min)
2
2.2
2.4
Ultra Trace Levels of Pesticides in Potable Water
Triazine herbicides analyzed at 50 parts per quadrillion
Atraton S/N =:: 819
6490 T
Triple
i l Q
Quad
d
with on-line enrichment
Simazine S/N = 18
Simetryn S/N = 9
Secbumeton S/N = 51
Prometon S/N = 40
Atrazine S/N = 28
Peptide MRM on 6460 vs new 6490 Triple Quad
x10
1
1 femtomole on 6460
4.5
LVNEVTEFAK
575.5 Æ 937.5
4.4
4.3
4.2
x10
10
4.5
4.4
4.3
4.2
1
100 attomole on 6490
Triggered MRM – What Problem Does tMRM Solve?
T
t d Quant
Q
t + Compound
C
d Identification
Id tifi ti
Targeted
Data dependent product ion scans leaves gaps in MRM
• mix
i off raw and
d synthetic
th ti points
i t
• higher RSDs and higher LOQs
Gap in MRM trace,
numerically filled in
in.
Typically > 100 msec
DDA –scan
switch to PIS
tMRM for up to 8 Confirmatory Ion Transitions
- when primary compound MRM triggers confirmatory MRMs
Faster compound ID at higher sensitivity with tMRM
Plot counts versus m/z for product ions
“Composite MS/MS spectra for compound b”
countts
QTRAP
5500
Vantage
Xevo
6490
100
200
300
Product
Ion Scan
Time
%MRM
y
Sensitivity
~ 100 msec
~ 10%
~ 150 msec
~ 150 msec
~ 2%
~ 2%
~ 30 msec
100%
The 6490 has Zeptomole Capability
what is a zeptomole?
p
Photo of new 6490 taken from 1 meter distance
How far away is 1021 meters?
The Milky Way Galaxy as seen from Hubble
1021 meters from the earth is at the edge of the known universe
Page 31
The New 6490 Triple Quad System
‰ New era in sensitivity – Zeptomole
‰ New era in linearity – Six decades of linearity
‰ New iFunnel Technology
‰ Improving compound identity - tMRMs
“Delivers Unprecedented Quantitative Excellence”