california tof/qtof users meetings: jim lau, agilent ms ... · nebulizer pressure drying gas...

California TOF/QTOF

Users Meetings:

MS Settings

Jim Lau, Agilent

Technologies.

Agilent Users Meeting

September 17th, 2008

Data Acquisition Overview

How to Improve Performance

• Optimize Source Parameters:

– Drying Gas Temperature, Gas Flow, and Neb - LC Condition and

Compound Dependent

– Fragmenter Voltage - Compound Dependent

• Optimize Ion Transmission in over the Mass Range

– QTOF For Low Mass Analytes < 200 Change Quad Amu Gain to 100

– TOF/QTOF Fragmentor

• Review How to Calibrate/Tune Q(TOF)

• Optimize Acquisition – Scan the Proper Mass Range

– Saturation and Acquisition Mode

– Set the Proper Acquisition Rate for Chromatographic Peaks

Agilent Users Meeting

September 17th, 2008

Optimization of ESI Source Parameters non-Ion

Funnel TOF/QTOF

VCap – Entrance of the Capillary, 2000 to 4500V / 5000VMAb

(3500V) Pos, (3000V) Neg

Dependent on compound and MW Concentrated samples (µg/µL) lower VCap

Drying Gas - Dependent on LC flow rate 10-13 L/min

Gas Temp - Start at high temperature work down-usually 275-325C

Skimmer - Voltage on skimmer and lens after capillary 65V

Fragmentor- Not Same as QQQ/Quad, Compound Dependent

Vary fragmentor/scan (30V steps) 100V to 240V

FIA with Injection Program

Sheath Gas- Dependent on LC flow rate 10-12 L/min

Shields Up!

Dual ESI Source Schematic

Vcap

Fragmentor

Nebulizer Pressure

Drying Gas Temperature and Flow

Drying Gas Flow high water needs higher flow if too low, spikes in spectra from droplets, dirty cap Drying Gas Temperature higher for low vapor pressure solvents Usually 275 - 350C Vcap optimize with FIA (2000-6000) start with 3000 V in negative mode, look for high chamber current or

blue glow (indicates corona): reduce Vcap if this happens

Electrospray Spray Chamber Settings

HPLC

Flow Rate

(µ/min)

Nebulizer

Pressure

(psi)

Drying Gas

Flow

(l/min)

Drying Gas

Temp

(C)

1 – 10 10 – 15 7 150 - 250

10 – 50 15 – 20 8 - 10 150 - 250

50 – 200 20 – 40 10 - 12 200 - 325

200 – 500 25 – 45 12 325

500 – 1000 50 – 60 12 - 13 350

Vcap

Corona

current

Nebulizer

Pressure

Fragmentor

Drying gas

Temperature

and Flow

Heater

APCI Spray Chamber Settings

HPLC

Flow Rate

(µl/min)

Nebulizer

Pressure

(psi)

Drying Gas

Flow

(l/min)

Drying Gas

Temp

(oC)

200 – 500 30-40 5 - 8 300-350

500 - 1000 40-60 8*-13* 350

APCI

Vap Temp

(oC)

Corona Current

(nA)

Capillary

Voltage

(V)

300 – 500 4000 (pos) & 20,000 (neg) 3500

* Use enough dry gas, but don’t remove reagent

Overview of the Multimode Source

Capillary

HPLC inlet

Nebulizer

Drying gas

Corona needle

ESI Zone

APCI Zone

Thermal container

To convert a method developed with any other source, all that is needed is to go to method and run control and change the Method spray chamber to MM-ES+APCI. Spray chamber values will be the same as for APCI

IR

Multimode Spray Chamber Settings

Mode Nebulizer

Pressure

(psi)

Drying Gas

Flow

(l/min)

Drying Gas

Temp

(oC)

APCI 30-60 5 - 8 300-350

ESI 30-60 8-13 325

APCI/ESI

Vap Temp

(oC)

Corona Current

APCI

(nA)

Capillary

Voltage

(V)

150(ESI)

250(APCI)

4000 (pos) & 20,000 (neg) 3500

Capillary

HPLC inlet

Nebulizer

Drying gas

Corona needle

ESI Zone

APCI Zone

Thermal container

Agilent Jetstream Technology (AJT) source

Non-ion funnel small molecule

The super-heated sheath gas collimates the nebulizer spray and presents more ions to the MS inlet.

*Nozzle voltage

Resistive sampling capillary exit: FragV

Nebulizing gas: pressure

*Sheath gas: flow and temperature

Drying gas:

flow and temperature

Resistive sampling capillary entrance: Capillary V

12

*New parameters unique to AJT source

Parameter

Starting Conditions

Agilent Jetstream

Pos/Neg

Nebulizer pressure 35-45 psi

Drying gas flow 10-12 LPM

Drying gas temp 275°C/325°C

Sheath gas flow 10-12 LPM

Capillary voltage 3500V/3000V

Nozzle voltage (AJT) 0V/1500V

Pos - low/Neg - higher

Fragmentor voltage

(cpd-dependent)

100-175V QTOF/TOF

Drying Gas Experiment on Agilent Jet Stream

Source 5l/min


Source 8l/min


Source 10l/min


Source at 12l/min

Drying Gas Experiment on Agilent Jet Stream Source

12l/min Note: a Higher Vcap increases High m/z response


Non-ion funnel intact protein


*Nozzle voltage

Resistive sampling capillary exit: FragV



Drying gas:



18


Parameter

Starting Conditions

Agilent Jetstream

Pos/Neg

Nebulizer pressure 55 psi

Drying gas flow 10 LPM

Drying gas temp 350°C


Capillary voltage 4500-5500V

Nozzle voltage (AJT) 2000V

Fragmentor voltage

(cpd-dependent)

225-400V QTOF, TOF

Note that “quad amu” will need to be increased 100-250amu with old quad drive in quad tuning parameters for QTOF


Ion funnel intact protein


*Nozzle voltage

Resistive sampling capillary exit:



Drying gas:



19


Parameter

Starting Conditions

Agilent Jetstream

Pos/Neg

Nebulizer pressure 55 psi

Drying gas flow 15LPM

Drying gas temp 250°C

Sheath gas flow 12 LPM

Sheath gas temp 250-300

Capillary voltage 4500-5500V

Nozzle voltage (AJT) 2000V

Fragmentor voltage

(cpd-dependent)

NA

Note that “quad amu” will need to be increased 100-250amu with old quad drive

in quad tuning parameters for QTOF


Ion funnel small molecule


*Nozzle voltage

Resistive sampling capillary exit:



Drying gas:



20


Parameter

Starting Conditions

Agilent Jetstream

Pos/Neg


Drying gas flow 15-17LPM

Drying gas temp 200-250°C


Sheath gas temp 200-400°C


Nozzle voltage (AJT) 0-500V/1500-2000V

Fragmentor voltage

(cpd-dependent)

NA

APCI source

Ion funnel small molecule

21

Parameter

Starting Conditions

Agilent Jetstream

Pos/Neg


Drying gas flow 14-15LPM

Drying gas temp 150-200°C

Vap Temp 350-400°C

Sheath gas temp NA


Chamber current 35 µa

Fragmentor voltage

(cpd-dependent)

NA

Vcap

Corona

current

Nebulizer

Pressure

Fragmentor

Drying gas

Temperature

and Flow

Heater

Ion Funnel Settings for Different Applications

6550 Tune (default)

6550 Tune

Peptide/Protein

6550Tune very

small molecule

(m/z <120) and

fragile adduct

6550 Generic

small molecule

6490 Tune

(default)

6490 Tune

generic small

molecule

6490 Tune Fragile

Compound and

Adduct

Funnel DC 50 50 30 50 15 15 15

HP Funnel Voltage Drop 150 150 100 150 180 180 130

HP Rf Voltage 200 200 90 150 200 200 150

LP Funnel Voltage Drop 100 100 50 100 100 100 90

LP Rf Voltage 100 100 40 60 110 110 40

Fragmenter to 175 Fragmentor to 325

Quad amu 100 Quad amu 120

These are suggested starting funnel parameters and will need optimization. Settings can be very analyte specific. They can be used “as is” should a user need to find out very quickly if it is worth the time to optimize the parameters further. These parameters may not be the same as listed from other sources that have been optimized for a particular analyte or class of analytes.

california tof/qtof users meetings: jim lau, agilent ms ... · nebulizer pressure drying gas...

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