breaking the femtogram sensitivity barrier with agilent jet

Breaking the Femtogram Breaking the Femtogram Sensitivity Barrier with Sensitivity Barrier with Agilent Jet Stream TechnologyAgilent Jet Stream Technology

TanayTanay WaingankarWaingankarMarketing Program Manager Marketing Program Manager -- LSSULSSU

What all do we need from an “ideal” Mass Analyser ….

� Wide Mass Range

� High Sensitivity

� High Resolution

� High Mass Accuracy

� True MS/MS capabilities

� Wide Linear Dynamic range

� Multiple Charge separation capabilities

� Polarity switching capabilities (Rapid or Slow)

� Complimentary Ionizations (ESI / APCI / APPI / MALD I )

� Quantitative analysis (MRM / PIS / NLS)

� Qualitative analysis (Multiple MS/MS capabilities)

All mass spectrometers sort ions based on their mass-to-charge ratios (m/z) in a vacuum

Common analyzer types:

� Quadrupole

�Single Quadrupole

�Triple Quadrupole

� Ion trap

� Time of flight

� Magnetic and electric sector

� Fourier transform ion cyclotron resonance

� Hybrid of any of the above mass analyzer

Mass Analyzers

LC- MS Components� Sample introduction

Direct Infusion, FIA, HPLC� Sample ionization

Nebulizer System� Ion transfer to high vacuum region

API Interface� Ion mass-to-charge filtering

Mass Analyzer� Ion detection

Detector� Data acquisition and analysis

Data System

API Inlet Analytical Domains

ElectroSpray Inlet (Ion Evaporation)

Electrospray (Pneumatically Assisted Electrospray)

Patented spray nebulizerPatented spray nebulizer

Patented orthogonal sprayPatented orthogonal spray

�� No adjustmentsNo adjustments�� Grounded spray needle for LC & CEGrounded spray needle for LC & CE�� Tolerant to nonvolatile ComponentsTolerant to nonvolatile Components

First and Best Orthogonal API Source Design

High Capacity Drying gas stream

Versatile sources

HPLC inlet

Nebulizer

Nebulizer gas inlet

Analyte IonsAnalyte Ions

WasteWasteWasteWasteConvenient drain port

Improved spectral quality & sensitivityClean capillary & ion optics

Orthogonal Source conceptOrthogonal Source conceptOrthogonal Source conceptOrthogonal Source conceptOrthogonal Source conceptOrthogonal Source conceptOrthogonal Source conceptOrthogonal Source concept-------- first first first first first first first first introduced by Agilent.introduced by Agilent.introduced by Agilent.introduced by Agilent.introduced by Agilent.introduced by Agilent.introduced by Agilent.introduced by Agilent.

Agilent Sprayer Design

Spray Droplet Size

Agilent LC/MSD

Some competitive designs

Signal

Our patented sprayer design leads to smaller

droplets and better sensitivity

LC/MS Ion Sources•Widest choice of sourcesfrom any manufacturer

– Electrospray (ESI)– Nnanospray– APCI– APPI– CE-TOF– PDF-MALDI

•Interchangeable between MS platforms

•Multimode Source

– ESI and APCI combined in one source

– Simultaneous operation•HPLC-Chip/MS

– Nanospray made ease– Reproducible results– Sample processing on chip

Ionize Anything!

Heated Nitrogen Drying gas protects Ion optics

Easy on labile compounds

Declustring

Droplet evaporation

Collision Induced Dissociation

Keeps non-ionizedspecies out of the analyzer

Heated nitrogen drying gas Interface

VACUUM

ATMOSPHERE

Different Ion Sampling Approaches

Broad Mass Range Transmission and Transmission Efficiency

0

0.2

0.4

0.6

0.8

1

1.2

0 500 1000 1500

Tra

nsm

issi

on .

Mass (m/z)

Mass Range TransmissionQuadrupole Hexapole Octopole

RF Ion Guide

Ion beam diverges behind Skimmer

RF Ion Guide preventsloss of ions andFocuses them intoMass analyzer

Ion transmissionEfficiency over widemass range

Design Of Various Mass Analyzer

Agilent Jet Stream Ion Generation

Nozzle voltage

MS inlet

Resistive samplingcapillary

Nebulizing gas

Super-heated sheath gas

Heated drying gas

LC sample inlet

The collimated thermal containment zone creates a dramatically “brighter source”

No ion left behind…

WHAT DOES JET STREM TECHNOLOGY PROVIDE A USER…

This technology � Enhances nebulization and desolvation � Reduces ion dispersion. �Delivers many more ions to mass analyzer.�Reduces number of neutral solvent clustures.

The result is higher signals with lower RSD at good limit of detection.

Agilent Jet Stream In ActionObserving Thermal Focusing

6/2/2009

Relentless Innovation

Less light scattering shows increased droplet desolvation and high ion production .

Agilent Jet Stream In ActionObserving Thermal Focusing

6/2/2009


Agilent systems are utilized worldwide to meet tough food testing regulatory standards for a broad range of compounds, including:

Range of CompoundsPesticidesAcrylamideAnti-Bacterial DrugsAnti-Viral Nucleoside CompoundsN-NitrosaminesBeta-AgonistsNitrofurane AntibioticsChloramphenicol and its MetabolitesSudan Food DyesPerfluorinated Organic Acids

Month ##, 200XPage 22

Agilent 6410 ESI vs 6460 Agilent ESI Jetstream

Negative ions~10x enhanced

6410ESI

Agilent Jet stream

6410ESI

Agilent Jet stream

Clorsulon50 ppb

Chloramphenicol50 ppb

For the treatment and control of internal parasites

Antimicrobial antibiotic

Agilent Jet Stream PerformanceSignal Enhancement – 6460 QQQ

PesticideOptimal

Temp [°C]Signal Gain

Alar 380 5.60

Acephate 380 6.30

Oxamyl 250 5.10

Carbendazim 380 7.20

Thiabendazole 380 7.20

Carbaryl 380 6.60

Imazalil 380 8.10

Methidathion 250 5.00

Chlorpyrifos-methyl

100 6.40

Pirimiphos-methyl 380 5.50

Chlorpyrifos 380 4.00

Trans-permethrin 250 2.50

Average 5.8x

12 Pesticides

Relative increase in signal of Agilent Jet Stream vs ESI = 5.8x

6x10

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1 2 3 4 5 6 7 8 9 10 11

alar

acephate

oxamyl

carbendazim

thiabendazolecarbaryl

imazalilmethidathion

Chlorpyrifos-methylPirimiphos-methylChlorpyrifos

trans-permethrin

Same InjectionSame Injection: : StdsStds in Solvent in Solvent --1uL injection @ 50ppt level (50 1uL injection @ 50ppt level (50 fgfg onon--column)column)

JetstreJetstreamamResultResultss

3x10

0.10.2

0.30.4

0.5

0.60.7

0.80.9

1+ TIC MRM (** -> **) 50ppt-r001.d 1 1 2 2 3 3 4 4 5 5

Counts vs. Acquisition Time (min)0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5 6 6.5 7 7.5 8 8.5 9 9.5 10 10.5 11 11.5 12 12.5 13 13.5 14 14.5 15

Agilent 6460 Agilent 6460 LC/MS/MSLC/MS/MS

Simazine

Prometon

Atrazine

Ametryn

Terbutryn

PrometrynPromazine

Agilent 6460 QQQTrace analysis of Triazine pesticides

Agilent 6460 QQQ PerformanceShattering the Femtogram Barrier – 500 Attograms

1.E+00

1.E+01

1.E+02

1.E+03

1.E+04

1.E+05

1.E+06

1.E+07

0 1 10 100 1000 10000 100000

Amount, fg

Pea

k A

rea,

cou

nts

R2 = 0.999

femtogram barrier

Five Decades of Linearity6460 triple quad with Agilent Jet Stream technology exhibits outstanding performance with 5 decades of linearity from sub-femtogram to 100 picograms of ve rapamil injected on-column.

Verapamil

1.8 µm C18, 2.1 x 50mm0.4 mL/min

500 attogram

Breakthrough Sensitivity500 Attograms verapamil on-column

Note: 3 x RMS

5 pg injections of Alprazolam in human plasma extr act

0

50000

100000

150000

200000

250000

300000

0 50 100 150 200 250 300 350 400 450 500

Injection Number

Are

a R

espo

nse

0.0

0.2

0.4

0.6

0.8

1.0

1.2

1.4

Rel

ativ

e R

espo

nse

Agilent Jet Stream PerformanceRuggedness & Reproducibility – 6460 QQQ

External Standard = 3.76% RSD

Internal Standard = 1.45 % RSD

500 Injections of Alprazolam in Spiked Human Plasma Extract, ~ 10hrs.

5x10

00.050.10.150.20.250.30.350.40.450.50.550.60.650.70.750.80.850.90.9511.051.11.151.21.251.31.351.41.451.51.551.61.65

Abundance vs. Acquisition Time (min)123456789101112131415161718192021222324252627282930313233

+ MRM (282.0 -> 212.0) mix100_100pg_5May.d 1 1 2 25x10

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

1

1.1

1.2

1.3

1.4

1.5

1.6

Abundance vs. Acquisition Time (min)1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33

1 1 2 2

Quantification of Multiple Targets

Issue: when trying to measure many targets in one analysis, the reduced time per MRM impacts sensitivity

Solution: Scheduled MRMs

6/2/2009


Scheduled MRMs

User inputs retention time and a time window for each target

System creates a table indicating which MRMs are needed at any given time

Dwell times adjusted to get even spaced points across signal for better integration results (constant cycle time). Can change every few seconds

6/2/2009


THANK YOU

Dr N Bhuvaneswaran – Application [email protected]

Nibin Varghese - [email protected][email protected] Free No: 1800113037

breaking the femtogram sensitivity barrier with agilent jet

Documents