Development ofthe World’s Mostthe World s Most

SensitiveTriple Quad MS

LC/MS Divisionand Agilent LabsAgilent TechnologiesTriple Quad MS Santa Clara, CA

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

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

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

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?

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.

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

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

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

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.

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

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)

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

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

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.

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

New 6490 LC Triple with iFunnel Technology “Unrivaled Quantitative Excellence”Unrivaled Quantitative Excellence

1290 Infinity LC/6490 Triple : Clearly Better Together

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

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

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]

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

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

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)

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

Page 24

q [ ]

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)

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

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

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

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

The 6490 has Zeptomole Capabilitywhat is a zeptomole?p

Photo of new 6490 taken from 1 meter distanceHow far away is 1021 meters?

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

Page 31

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”