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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
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
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
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.
Less light scattering shows increased droplet desolvation and high ion production .
Agilent Jet Stream In ActionObserving Thermal Focusing
6/2/2009
Relentless Innovation
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
Page 22 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
Page 25
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
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
Relentless Innovation
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
Relentless Innovation
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