2
Agenda 1.Introducción
2.Resumen de tecnologías disponibles en LCMS y GCMS
3.Tecnología i-funnel, revolucionamos el muestreo de iones para alcanzar máxima
sensibilidad
4.Análisis target con LCQQQ
5.Análisis de desconocidos con sistemas de tiempo de vuelo
Introducción:Necesidades y limitaciones Como de crítica es la preparación de la muestra?
Tiempo, dinero
Lots of effort:
Full scan MS/MS SIM where
appropriate
The goal is to get reliable data
Or to get to the Mountain top
Agilent GC/MS More Choices – Better Solutions
240 IT 7890 GC
7000 TQ 7890 GC
5975C SQ 7890 GC
220 IT 7890 GC
5975E SQ 7820 GC
5975T LTM SQ
7200 Q-TOF 7890 GC
Tecnologías GCMS
Page 11
“…sólamente somos capaces de detectar
1 de cada 103 a 105 iones
generados por electrospray (ESI) a presión
atmosférica con los diseños instrumentales
actuales.”
(Page, Smith, et al; JASMS, 2007, 18, 1582-1590)
Tecnología i-Funnel
Agilent Jet Stream
• Confinamiento térmico
de la pluma de ESI
• Desolvatación eficaz
para crear iones en fase
gas
• Genera una zona
enriquecida de iones
N E W I F U N N E L T E C H N O L O G Y
Hexabore Capillary
• 6 capilares de entrada
• Las muestras son 10X
más concentradas a partir
de la fuente
• Captura la mayoría dels
gas que proviene de la
región de la fuente ESI
Dual Ion Funnel
• Elimina el gas pero
captura los iones
• Elimina el ruido de
neutros
• Prolonga la vida de la
bomba turbomolecular
La tecnología iFunnel captura 10X más iones
Page 12
Page 13
Nuevo Triple cuadrupolo 6490 “The highest sensitivity triple quadrupole on the market”
JP Morgan Healthcare Conf. 2011
Sensibilidad de Zeptomotles
• microdosing
• dried blood spots
• hormone analysis
• personal care products in water
• pesticides – dilute and shoot
Dynamic Range of Six Orders Linearity eliminates serial dilution of samples
analyze trace level impurities in one run
Triggered MRM (tMRM)
library searching – NIST data base fit scores
create your own data base for searches
6550 iFunnel Q-TOF
LC/MS System
ASMS 2010
Sensitivity
• Dramatically improved quantitative capabilities
• New Qual/Quan Workflows
• Superior metabolite and protein detection
• Non-targeted compound screening
Comprehensive Performance Enhancements
• Mass Resolution >40,000
• 50 spectra /sec MS and 33 spectra/sec MS/MS
• 5 orders of linear dynamic range
• <1 ppm MS mass accuracy; <2 ppm MS/MS
• Unrivalled sensitivity
Nuevo 6550 iFunnel QTOF
10X Sensitivity Gain Enables Applications
Compound Detection Below 10 ppb
Pepper Pepper Orange Orange
Compound 6530 6550 6530 6550
Chlorfluazuron
Cymoxanil
Diuron
Fluazifop-butyl
Fluroxypr
Propaquizafop
Quizalofop-ethyl
Rotenone
Bromacil
Fenproximate
Tribenuron
Aldoxicarb
85% of all pesticides successfully
detected by 6530 QTOF in
tomato, pepper, leek, orange
matrices
Majority of Low Responders
Detected by 6550
New Agilent LC Triple Quad Every Year
Since 2007
6460
Triple
Quad
6430
Triple
Quad
6420
Triple
Quad
6490
Triple
Quad
6460
Triple
Quad
Pe
rfo
rma
nc
e
Análisis target con LCQQQ
Upgradeability…
Infinity LC
6420
Easy Choice – Mid-range markets
Routine Trace Analyses
6430
6460
Perf
orm
an
ce
Routine Quantitation
iFunnel
Technology
Ultra-trace & fast
Analyses
6490
Dynamic MRM Powerful Method for Multi-Analyte Analysis
• MRM con ventanas de tiempo de retención
• Menos MRMs concurrentes
• Dwell Times mayores
• Menores LODs
• Menores RSDs
• Fixed Cycle, Variable Dwell:
Constant Cycle Time Assures
Accurate Quantitation
Counts vs. Acquisition Time (min)
x105
1
2
3
4
5
1 2 3 4 5 6 7 8 9 10 11 12 13 14
Segmento 1 Segmento 2
Segmento 3
Segmento 4
Análisis por segmentos vs. MRM dinámico
MRM Dinámico: No hay segmentos de tiempo La lista de MRM se confecciona de modo dinámico basándose en el RT de los compuestos de interés
Menor tiempo de ciclo/Mejor definición de pico
Page 20
Agilent 1290 Infinity LC + 6460A Triple Quad with Agilent Jet Stream Technology
~600 Dynamic MRM Transitions
9 replicates analyzed at 200 pg level
Mean area %RSD = 3.2, Mean height %RSD= 3.6
Application: Analysis of 300 Pesticides in 15 min.
The combination of UPLC-QQQ with Dynamic MRM acquisition results in high sensitivity,
quantitative data and very fast analysis times
152.1
77.0
105.0 219.0
m/z
60 100 140 180 220
209.1
165.1
triggered MRM Spectra
Triggered MRM (tMRM) – a Data Dependent Scan Data dependent execution of additional, confirmatory MRMs
2-Ethylanthraquinone
Triggered MRM in MassHunter Acquisition
Threshold
Additional MRMs used
for compound confirmation
Primary cycle (below threshold)
Triggered cycle (above threshold)
tMRM Product Ion Spectrum on any 6400 Triple A Re-Constructed Product Ion Spectrum that is Library Searchable
x103
8
7
6
5
4
3
2
1
0 80 100 120 140 160 180 200 220 240 260 280 300 320 340
334.2 171.1
145.0
132.1
117.0
105.1 91.1 76.0
tMRM Product Ion Spectrum
Tebufenpyrad
x103 8
7
6
5
4
3
2
1
0 80
76.0
x103 8
7
6
5
4
3
2
1
0 80 100
91.1
x103 8
7
6
5
4
3
2
1
0 80 100 120
105.1
x103 8
7
6
5
4
3
2
1
0 80 100 120
117.0
x103 8
7
6
5
4
3
2
1
0 100 140
132.1
x103 8
7
6
5
4
3
2
1
0 80 100 120
119.0
x103 8
7
6
5
4
3
2
1
0 100 140 180
145.0 x103
8
7
6
5
4
3
2
1
0 100 140 180
147.0
x103 8
7
6
5
4
3
2
1
0 80 100 120 140 160 180
171.1
334 > 76 334 > 91 334 > 105 334 > 117 334 > 119
334 > 132 334 > 145 334 > 147 334 > 171
Acquisition Time (min)
1 1.2 1.4 1.6 1.8 2 2.2 2.4 2.6 2.8 3 3.2 3.4 3.6 3.8 4 4.2
Amphetamine
Metham-
phetamine
Trazodone
PCP
Norver-
apamil
Verapamil
Methadone
Proadifen
Benzoyl-
ecgonine
Cocaine
Forensics Mixture with 6420 and tMRM Method
Triggered MRM actuated
on red peak
@ 5ppb level:
RSD=4.9% (N=3)
Triggered MRM for Compound Confirmation
6420 tMRM: Benzoylecgonine, 25 pg on column
Mass-to-Charge (m/z)
100 150 200 250 300 -1.2
-1
-0.8
-0.6
-0.4
-0.2
0
0.2
0.4 0.6
0.8
1
168.1
168.1
77.0
77.0 105.0
105.0
290.1
Ref. Library Match Score = 95.1
Peak Product
Ion Spectra
Library
Reference
Product Ion
Spectra
Top Related