Development of

the World’s
World s Most LC/MS Division
Sensitive and Agilent Labs
Agilent Technologies
Triple Quad MS 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

Hexabore Capillary Dual Ion Funnel

• Thermal confinement • 6 capillary inlets • Removes the gas but

of ESI plume captures the ions
• Samples 6X more ion rich
• Efficient desolvation to gas from the source • Removes neutral noise
create gas phase ions
• Captures the majority of • Extends turbo pump life
• Creates an ion rich zone the gas from the source
region

3
Agilent Jet Stream Ion Generation

Gas Dynamics View

N b li i gas
Nebulizing
Enhanced efficiency nebulizer

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” Resistive sampling
capillary
Patent Pending
Agilent’s Hexabore Atmospheric Sampling….
The Most Sensitive…. By
y Far.
6 Bore
Capillary

• Nearly 6 x the amount of

p
atmospheric gas
g sampled
p
compared to a single
capillary system
AND

• 5 - 10x more ions into

g vacuum region
the high g

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 LLow
Pressure Pressure
Stage 1 Stage 2

Existing
RF Ion
Guide

Stage 2
Stage 1
St 1-3 Torr
8-12 Torr

Stage 1 offset breaks up the high pressure gas exiting the Hexabore Capillary

8
How Does an Ion Funnel Work?
The RF Voltage RF Drive
focuses the ions
to the center.

Unfocused
Ions and Gas Focused
Enter Ions Out

The DC Voltage
accelerates the
ions to the exit. DC Drive
How is an Ion Funnel Constructed?
Gold Plated
Previously, with many metal plates…. Rim

… 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
Improved Ion Sampling Efficiencies with iFunnel

Agilent Jet Stream Agilent Jet Stream

positive ion negative ion

7x 11x
7x
6490 14x
7x
7x 5x 10x
5x
10x

118 622 922 6460

322 302 1034
1222

113

200 400 600 800 1000 1200 1400 200 400 600 800 1000 1200 1400
m/z m/z
 6490 with iFunnel vs 6460 in positive ion mode

1 pg Reserpine (7X) 1 pg Alprazolam (6X)

103
x10 x103
6490
6490

1
6460
6460

0 0.1 0.2 0.3 0.4 0.5 0.6 0 0.2 0.4 0.6 0.8 1.0 1.2
A
Acquisition
i iti Time
Ti (min)
( i ) Acquisition Time (min)

12
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 6490
S/N = 1,157:1 S/N = 11,640:1
where noise = 1xRMS where noise = 1xRMS
 6490 with iFunnel vs 6460 in negative ion mode

1 pg Chloramphenicol (8X) 1 pg Acid red 4 (9X)

x103 x103
3
3
6490 1.1 6490
1
Relative
Increase in 0.9
2 signal of T it
Triton
0.8
Triton vs. 6460
6x 0.7
0.6
0.5
1 0.4
6460 BB1: 0.3
CAT CC
only
6460 0.2
6460
0.1
0 0
0.5
0 5 1 15
1.5 2 0.2
0 2 0.4
0 4 0.60 6 0.8
08 1
Acquisition Time (min) Acquisition Time (min)

14
 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)) Larger
(5 mm)
Exit Entrance
Aperture
Focuses Aperture
I
Ions iinto
t Accepts more
MS2 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
10000
9000
SNR 1 pg reserrpine

8000
7000
6000
5000
4000
6460
3000
2000
6430
6410
1000
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
100 Attogram to 100 Picogram varapmil
arapmil on col mn
on-column
10000000
Y = 65283 x – 0.390412
1000000 R2 = 0.9976
0 9976

100000
nse
ea Respon

10000

1000
Are

100

10

1
0.1 1 10 100 1000 10000 100000

Femtograms
e tog a s
 50 pg/mL Fluticasone in plasma with 6460

25 Femtogram on-column
Area RSD = 23.7% (n=6)
Mean accuracy = 104
Fluticasone propionate
F
x10 1 *1.534 min.
S O
5.3 CH3
CH3 O
5.2 HO
O
CH3 CH3
5.1
5 F H

4.9 O
F
4.8
4.7
4.6
4.5
4.4 MRM unit / unit resolution
4.3 (0.7 0.7
(0 7 amu / 0 7 amu)
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 1.9 2 2.1 2.2 2.3 2.4

Acquisition Time (min)

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
Fluticasone i t
propionate
1.765 min.
x10 1
F
x10 1
5.3 5.3 S O CH3
5.2 5.2 HO
CH3 O MRM enhanced / unit resolution
51
5.1 5.1 CH3
O
CH3 (0 4 amu / 0.7
(0.4 0 7 amu)
5 5 F H
4.9 4.9 O

4.8 4.8 F

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

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)

Enhanced Resolution gives increased selectivity for

1 analytes in complex matrices
x10

Q1 Unit (0.7 amu)

7
Q2 Unit (0.7 amu)

Q1 Enhanced (0.4 amu)

6
Q2 Unit (0.7 amu)

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)

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 QQuad
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
1
x10 1 femtomole on 6460
4.5
LVNEVTEFAK
4.4 575.5 Æ 937.5

4.3

4.2

1
x10
10
100 attomole on 6490
4.5

4.4

4.3

4.2
Triggered MRM – What Problem Does tMRM Solve?

T t d Quant
Targeted Q t + Compound
C d Identification
Id tifi ti

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”
Product %MRM
Ion Scan y
Sensitivity
Time
QTRAP
~ 100 msec ~ 10%
5500
countts

Vantage ~ 150 msec ~ 2%

Xevo ~ 150 msec ~ 2%
6490 ~ 30 msec 100%

100 200 300

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”