HESI-II Probe User Guide

HESI-II Probe User Guide
HESI-II Probe
User Guide
70005-97000 Revision C
April 2009
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C
Contents
Preface . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Safety and Special Notices . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii
Precautions for Handling the HESI-II Probe . . . . . . . . . . . . . . . . . . . . . . . . . .viii
Contacting Us . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .ix
Chapter 1
Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1
Chapter 2
Functional Description. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .5
Chapter 3
Removing and Installing the HESI-II Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9
Removing the HESI-II Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Installing the HESI-II Probe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
Adjusting the Probe Position . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17
Chapter 4
Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .19
Flushing the Sample Transfer Line, Sample Tube, and HESI-II Probe . . . . . . . 19
Replacing the Needle Insert. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20
Chapter 5
Replaceable Parts. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .23
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .25
Thermo Scientific
HESI-II Probe User Guide
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P
Preface
This HESI-II Probe User Guide provides you with information on using the
heated-electrospray ionization probe. It also provides procedures for installing and
maintaining the HESI-II probe.
Safety and Special Notices
Make sure you follow the precautionary statements presented in this guide. The safety and
other special notices appear in boxes.
Safety and special notices include the following:
CAUTION Highlights hazards to humans, property, or the environment. Each CAUTION
notice is accompanied by an appropriate CAUTION symbol.
IMPORTANT Highlights information necessary to prevent damage to software, loss of
data, or invalid test results; or might contain information that is critical for optimal
performance of the system.
Note Highlights information of general interest.
Tip Highlights helpful information that can make a task easier.
Thermo Scientific
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Preface
Precautions for Handling the HESI-II Probe
When operating the mass spectrometer in the heated-electrospray mode, do not touch the
heated surfaces of the HESI-II probe or the Ion Max API source housing. Touching the
heated surfaces of the probe or the source housing can cause burns when the probe is
operating at typical temperatures of 350 to 450 °C.
CAUTION AVOID BURNS. At typical operating temperatures (350 to 450 °C), the
HESI-II probe can severely burn you. Before removing the probe from the API source
housing, allow the probe to cool to room temperature (for approximately 20 minutes)
before you touch it.
When operating the mass spectrometer in the heated-electrospray mode, if you observe liquid
leaking from the sample inlet, place the mass spectrometer in Standby mode before you
tighten the fitting to eliminate the leak.
CAUTION AVOID ELECTRIC SHOCK. Do not tighten the probe sample inlet fitting
to eliminate a liquid leak while the mass spectrometer is in operation. If you touch liquid
leaking from the probe sample inlet while the mass spectrometer is in operation, you
might receive an electric shock.
A
S
Avoid touching liquid leaking from
the sample inlet while the mass
spectrometer is in operation.
General caution
Hot surface caution
Hot surface caution
viii
HESI-II Probe User Guide
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Preface
Contacting Us
There are several ways to contact Thermo Fisher Scientific for the information you need.
Y To contact Technical Support
Phone
800-532-4752
Fax
561-688-8736
E-mail
[email protected]
Knowledge base
www.thermokb.com
Find software updates and utilities to download at mssupport.thermo.com.
Y To contact Customer Service for ordering information
Phone
800-532-4752
Fax
561-688-8731
E-mail
[email protected]
Web site
www.thermo.com/ms
Y To copy manuals from the Internet
Go to mssupport.thermo.com and click Customer Manuals in the left margin of the
window.
Y To suggest changes to documentation or to Help
• Complete a brief survey about this document by clicking the link below. Thank you
in advance for your help.
• Send an e-mail message to the Technical Publications Editor at
[email protected]
Thermo Scientific
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1
Introduction
Heated-electrospray ionization (H-ESI) transforms ions in solution into ions in the gas phase
by using electrospray ionization (ESI) in combination with heated auxiliary gas. You can use
H-ESI to analyze any polar compound that makes a preformed ion in solution.
Basic compounds (for example amines) can form a protonated molecule [M + H]+, and acidic
compounds (for example sulphonic acids) can form a deprotonated molecule [M – H]-. In the
positive ion polarity mode, the protonated molecule produces a peak at an m/z value of
M + 1, where M equals the mass of the original molecule. In the negative ion polarity mode,
the deprotonated molecule produces a peak at an m/z value of M – 1, where M equals the
mass of the original molecule.
Because solution chemistry is the primary factor affecting mass spectra in ESI, other common
adducts include sodium ions (Na+), yielding an m/z value of [M + 23]+; potassium ions (K+),
yielding an m/z value of [M + 39]+; and ammonium ions (NH4+), yielding an m/z value of
[M + 18]+.
This chapter describes the principles of the heated-electrospray ionization technique through
use of the HESI-II probe (see Figure 1) and a Thermo Scientific mass spectrometer.
Figure 1.
Thermo Scientific
HESI-II probe
HESI-II Probe User Guide
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Introduction
With H-ESI, a mass spectrometer can analyze a range of molecular weights greater than
100 000 u, due to multiple charging. H-ESI is especially useful for the mass analysis of polar
compounds, which include biological polymers (for example, proteins, peptides,
glycoproteins, and nucleotides); pharmaceuticals and their metabolites; and industrial
polymers (for example, polyethylene glycols).
In H-ESI, ions are produced and introduced into the mass spectrometer as follows:
1. The sample solution enters the ESI needle, which receives a high voltage.
2. The ESI needle sprays the sample solution into a fine mist of droplets that are electrically
charged at their surface.
3. The electrical charge density at the surface of the droplets increases as solvent evaporates
from the droplets. In heated-electrospray ionization, heated auxiliary gas aids solvent
evaporation.
4. The electrical charge density at the surface of the droplets increases to a critical point
known as the Rayleigh stability limit. At this critical point, the droplets divide into
smaller droplets because the electrostatic repulsion is greater than the surface tension. The
process is repeated many times to form very small droplets.
5. Electrostatic repulsion ejects sample ions from the very small, highly charged droplets
into the gas phase.
6. The sample ions enter the mass spectrometer through the ion transfer tube.
Figure 2 shows the steps in the formation of ions from highly charged droplets.
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Figure 2.
Introduction
H-ESI process in the positive ion polarity mode
Ion transfer
tube
HESI-II probe
nozzle
–
–
+ + +
– +– +
+– – – +
+ – –+ +
+ +
Charged
droplet
++
+–
––
–
+ +–
+
+ –+
Solvent
evaporates
from droplet
+
+
Ejected
positive ions
You can use the HESI-II probe in either positive or negative ion polarity mode. The ion
polarity mode of choice is determined by the polarity of the preformed ions in solution:
Acidic molecules form negative ions in solution, and basic molecules form positive ions.
Because the ejection of sample ions from droplets is facilitated when the ionic charge and
surface charge of the droplet have the same polarity, use the positive ion polarity mode to
analyze positive ions and the negative ion polarity mode to analyze negative ions.
Sample ions can carry a single charge or multiple charges. The number of charges carried by
the sample ion depends on the structure of the analyte of interest and the carrier solvent.
(In H-ESI, the buffer and the buffer strength both have a noticeable effect on sensitivity, so it
is important to choose these variables correctly.) In the case of higher molecular weight
proteins or peptides, the resulting mass spectrum consists typically of a series of peaks
corresponding to a distribution of multiply charged analyte ions.
Droplet size, surface charge, liquid surface tension, solvent volatility, and ion solvation
strength are factors that affect the H-ESI process. Large droplets with high surface tension,
low volatility, strong ion solvation, low surface charge, and high conductivity prevent good
electrospray.
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Introduction
Organic solvents, such as methanol, acetonitrile, and isopropyl alcohol, are superior to water
for H-ESI. Volatile acids and bases are good, but salts above 10 mM concentration and strong
acids and bases are extremely detrimental.
Follow these rules for achieving a good electrospray:
• Keep salts out of the solvent system.
• Use organic/aqueous solvent systems and volatile acids and bases.
• Optimize the pH of the solvent system.
Table 1 shows initial heated electrospray settings for different liquid flow rates for a 50 percent
aqueous solution. These initial settings provide a starting point for optimizing system
performance. The optimal settings for your application depend on the compounds of interest,
the solvent matrix, and the chromatographic conditions. For information on optimizing these
settings, refer to the getting started guide for your mass spectrometer.
Tip For best results, avoid operating the HESI-II probe at elevated temperatures without
solvent flow from the LC system or the syringe pump. Allowing the HESI-II probe to run
dry at elevated temperatures can cause blockage of the replaceable metal needle (see
“Replacing the Needle Insert” on page 20).
For an LTQ Series mass spectrometer, the allowable range for the auxiliary gas flow depends
on the version of the instrument control software and the vaporizer temperature setting:
• For LTQ 2.5.0 or lower, the allowable range for the auxiliary gas flow is 5 to 60 units.
• For LTQ 2.5.5 or higher, the allowable range for the auxiliary gas flow is 0 to 60 units for
vaporizer temperatures up to 100.00 °C. For vaporizer temperatures above 100.00 °C, the
minimum auxiliary gas flow is 5 units.
For a TSQ Series mass spectrometer, you can set the auxiliary gas flow from 0 to 60 units.
Tip For best results, set the auxiliary gas flow to a minimum of 5 units for vaporizer
temperatures above 100.00 °C.
Table 1. Initial heated electrospray settings
*
4
Liquid
flow rate
(μL/min)
Ion transfer
tube
temperature
(°C)
H-ESI
vaporizer
temperature
(°C)
Sheath gas
pressure
(psi)
Auxiliary gas
flow
(arbitrary units)
Spray voltage
(V)
Typical
nitrogen gas
consumption
(L/min)
5
240
Off to 50
5
0
+3000 (-2500)*
<1
200
350
250 to 350
35
10
+3000 (-2500)
8
500
380
300 to 500
60
20
+3000 (-2500)
13
1000
400
500
75
20
+3000 (-2500)
17
Negative ion mode
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Functional Description
The HESI-II probe produces charged aerosol droplets that contain sample ions. The HESI-II
probe accommodates liquid flows of 1 μL/min to 1 mL/min without splitting.
The removable components of the HESI-II probe are the end cover that is secured to the
probe body with socket head screws and the needle insert that screws into the probe body
(see Figure 3). The end cover includes the high voltage feedthrough and the sample inlet port.
The external components of the probe body include the grounding union holder, vaporizer
cable connector socket, probe sleeve with depth markers, and probe nozzle (see Figure 3 and
Figure 4).
Figure 3.
HESI-II probe removable components
Probe body
End cover
Needle insert
Probe sleeve with
depth markers
Socket head screws
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Functional Description
Figure 4.
HESI-II probe views
Sheath gas inlet (S)
Probe nozzle
ESI needle
Aux gas inlet (A)
1.5 mm
Vaporizer cable
connector socket
Metal needle
High voltage socket for the
8 kV cable connector
Sample inlet
Grounding
union holder
Sample and solvent enter and exit the HESI-II probe through the needle insert, which
protrudes from the sample inlet port at the back of the probe and from the nozzle at the front
of the probe (see Figure 4). The needle insert (see Figure 3 on page 5) includes an adjustable
union, a needle guide fitting and ferrule, an ESI needle, and a metal needle. The adjustable
union is used to adjust the protrusion of the needle insert tip from the probe nozzle (see
Figure 4).
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Functional Description
A large negative or positive voltage is applied to the ESI needle (typically ±3 to ±5 kV), which
sprays the sample solution into a fine mist of charged droplets. The probe nozzle directs the
flow of sheath gas and auxiliary gas at the droplets. The probe body houses the probe nozzle
and needle insert, and includes the sheath gas and auxiliary gas plumbing.
The probe body has inlets for the sheath and auxiliary gases. The sheath gas is the inner
coaxial nitrogen gas that sprays (nebulizes) the sample solution into a fine mist as it exits the
metal needle sample tube (see Figure 5). The heated auxiliary gas is the outer coaxial nitrogen
gas that assists the sheath gas in the desolvation of sample solutions (see Figure 6). The
auxiliary gas heats as it passes through a vaporizer. The vaporizer is thermally insulated from
the sample tube to prevent direct heating of the sample solution. You can control the
vaporizer temperature from the Xcalibur™ data system. The temperature range is from
ambient room temperature to 600 °C. For recommended operating temperatures and gas flow
settings, see Table 1 on page 4.
Figure 5.
Sheath gas plumbing
Sheath gas fitting (blue)
in gas inlet (S)
Sheath
gas
ESI needle
(outer needle)
Metal needle
(inner needle)
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Functional Description
Figure 6.
Auxiliary gas plumbing
Auxiliary gas fitting (green)
in gas inlet (A)
Auxiliary gas
ESI needle
(outer needle)
Metal needle
(inner needle)
The angle of the HESI-II probe is fixed at approximately 60 degrees. To help optimize the
spray stability, use adjustment screws on the Ion Max source housing to make small changes to
probe position. The fixed angle, off-axis spraying affords long-term signal stability
(robustness) for most solutions that contain non-volatile matrix components, mobile phase
buffers, or ion-pairing reagents. For information on adjusting the probe position, see
“Adjusting the Probe Position” on page 17.
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Removing and Installing the HESI-II Probe
This chapter describes how to set up the API source with the HESI-II probe. You must
remove the HESI-II probe to perform maintenance on the probe.
Note To install the APCI probe, refer to the Ion Max and Ion Max-S API Source Hardware
Manual.
Contents
• Removing the HESI-II Probe
• Installing the HESI-II Probe
• Adjusting the Probe Position
Removing the HESI-II Probe
CAUTION AVOID BURNS. At typical operating temperatures (350 to 450 °C), the
HESI-II probe can severely burn you. Before removing the probe from the API source
housing, allow the probe to cool to room temperature (for approximately 20 minutes)
before you touch it.
Y To remove the HESI-II probe from the API source housing
1. Place the mass spectrometer in Standby mode, and allow the probe to cool to room
temperature.
Wait approximately 20 minutes for the probe to reach room temperature when it has
been operating at high temperature. If the mass spectrometer is connected to an LC
system, leave the solvent flow from the LC pump on while the probe is cooling to room
temperature.
2. If the mass spectrometer is connected to an LC system, turn off the solvent flow from the
LC pump, and then disconnect the tubing from the left side of the HESI-II probe
grounding union (see Figure 7 on page 10).
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Removing and Installing the HESI-II Probe
Removing the HESI-II Probe
Figure 7.
Front view of the connections to the HESI-II probe
Sheath gas line
8 kV cable
Vaporizer cable
Auxiliary gas line
To LC outlet
Connection between
grounding union and
sample inlet
Probe locking knob
3. Disconnect the 8 kV cable from the HESI-II probe high voltage receptacle as follows (see
Figure 8 and Figure 9):
a. Unlock the cable by twisting the locking ring counterclockwise.
b. Unplug the 8 kV cable from the HESI-II probe high voltage socket.
Figure 8.
Enlarged view of the 8 kV cable connector
Locking ring
High voltage socket
(for the 8 kV cable
connector)
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Removing and Installing the HESI-II Probe
Removing the HESI-II Probe
4. Unplug the vaporizer cable from the HESI-II probe vaporizer cable socket (see Figure 9
and Figure 10).
Figure 9.
Top view of the API source housing and the HESI-II probe
Vaporizer cable
(connected to
the probe)
8 kV cable
(disconnected)
A
S
Probe vaporizer
cable socket
5. Disconnect the auxiliary gas fitting (green) from the auxiliary gas inlet (A) on the probe
(see Figure 10).
6. Disconnect the sheath gas fitting (blue) from the sheath gas inlet (S) on the probe (see
Figure 10).
Figure 10. Top view showing the sheath and auxiliary gas lines connected to the probe
Vaporizer cable
(disconnected)
8 kV cable
(disconnected)
A
S
Sheath gas
line
Thermo Scientific
Auxiliary gas
line
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Removing and Installing the HESI-II Probe
Removing the HESI-II Probe
7. Connect the vaporizer cable to the interlock socket on the interlock block. To align the
socket pins, align the red dot on the vaporizer cable connector with the red dot on the
interlock socket (see Figure 11).
Figure 11. View of the left side of the API source housing
Vaporizer cable
Vaporizer cable
socket
Sheath gas inlet
Red dot on the
vaporizer cable
Interlock socket
8. Unlock the probe locking ring by turning the probe locking knob (see Figure 7 on
page 10) counterclockwise.
9. Remove the probe from the port in the API source housing as follows:
a. Slowly pull the probe out of the port until you feel the resistance caused by the probe
guide pin meeting the interlock block.
b. Turn the probe counterclockwise until the guide pin is free of the interlock block.
c. When the guide pin is free of the interlock block, pull the probe out of the port.
10. Store the HESI-II probe in its original shipping container.
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Removing and Installing the HESI-II Probe
Installing the HESI-II Probe
Installing the HESI-II Probe
Note To remove an APCI or ESI probe, refer to the Ion Max and Ion Max-S API Source
Hardware Manual.
Y To install the HESI-II probe
1. Remove the HESI-II probe from its storage container. Inspect and clean it if necessary.
Figure 12 shows the locations of the probe guide pin, vaporizer cable connector socket,
sheath and auxiliary gas inlets, 8 kV cable connector socket, sample inlet, and grounding
union holder.
Figure 12. HESI-II probe views
Vaporizer cable
connector socket
Guide pin
Auxiliary gas
inlet
8 kV cable
connector
socket
Sheath gas
inlet
Sample inlet
Grounding union holder
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Removing and Installing the HESI-II Probe
Installing the HESI-II Probe
2. Turn the probe locking knob counterclockwise until the probe locking ring is opened to
its widest position (see Figure 13).
Figure 13. Ion Max API source housing without a probe
Interlock socket
Probe interlock block
Probe locking knob
Probe locking ring
Probe port
3. Seat the probe in the probe port in the API source housing as follows:
a. Holding the probe with the nozzle facing downward and the guide pin facing toward
the left, slowly insert the probe into the port until the guide pin meets the locking
ring on the API source housing (see Figure 14).
Figure 14. View of the guide pin touching the locking ring
Slot in the left side of
the interlock block
Guide pin
Locking ring
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Removing and Installing the HESI-II Probe
Installing the HESI-II Probe
b. Pull the probe slightly upward until the guide pin is level with the slot on the left side
of the interlock block. Then turn the probe clockwise until the guide pin meets
resistance from the interlock block (see Figure 15).
Figure 15. View of the guide pin inserted into the slot
Guide pin inserted into the
slot in the left side of the
interlock block
A
C B
D
A, B, C, and D depth
markers
c. Push the probe further downward into the port to the appropriate depth indicated by
the A, B, C, and D depth markers on the probe.
In the H-ESI mode, insert the probe to a depth of B, C, or D. For high solvent flow
rates, adjust the probe depth so that the nozzle is farther away from the ion interface
(depth C or D). Conversely, for low solvent flow rates, adjust the probe depth so that
the nozzle is closer to the ion interface (depth B or C).
4. Lock the probe in place by turning the probe locking knob (see Figure 13 on page 14)
clockwise until you feel resistance.
5. Connect the nitrogen gas lines to the HESI-II probe as follows (see Figure 10 on
page 11):
a. Connect the sheath gas fitting (blue) to the sheath gas inlet (S).
b. Connect the auxiliary gas fitting (green) to the auxiliary gas inlet (A).
6. Unplug the vaporizer cable connector from the interlock socket and connect it to the
vaporizer cable connector socket on the HESI-II probe. To align the connector pins with
the socket, align the red dot on the connector with the red dot on the socket. Figure 11
on page 12 shows the red alignment dots on the connector and the socket.
7. Connect the 8 kV cable connector to the 8 kV cable connector socket on the HESI-II
probe. Tighten the locking ring (see Figure 8 on page 10) on the 8 kV cable connector.
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Removing and Installing the HESI-II Probe
Installing the HESI-II Probe
8. Ensure that the grounding union (stainless steel ZDV fitting) is seated in the grounding
union holder on the HESI-II probe.
9. Using two fingertight fittings, connect a short length of red PEEK tubing to the right side
of the grounding union and to the probe sample inlet.
10. Using two fingertight fittings, connect a length of red PEEK tubing to the LC outlet and
to the left side of the grounding union.
Figure 16 shows the HESI-II probe installed in the Ion Max API source.
Figure 16. HESI-II probe installed in the Ion Max API source housing
Vaporizer cable
Sheath gas line
(blue fitting)
Auxiliary gas line
(green fitting)
A
S
8 kV cable
Sample inlet
To the LC system
Grounding union holder with a
stainless steel grounding union
Tip Before analyzing samples, change the ion source mode in TSQ EZ Tune by choosing
Setup > Change Ion Source > HESI.
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Removing and Installing the HESI-II Probe
Adjusting the Probe Position
Adjusting the Probe Position
To maximize sensitivity, you can adjust the probe depth. If you have the Ion Max API source
housing, you can also adjust the side-to-side and front-to-back probe position by a few
millimeters.
For best results, follow the guidelines in Table 2 as a starting point. These guidelines are based
on the liquid flow rate of your application.
Table 2. Probe position guidelines
*
Liquid flow rate range
(μL/min)
Front-to-back
position
(micrometer setting)
Probe depth
(probe depth line)
Side-to-side
position
(+1 to -1 marks)
1 to 50
1.75*
B
0
50 to 2000
1.75
C
0
Fully turned to the right
Y To adjust the probe position
• Using the micrometer on the front of the Ion Max source housing, adjust the
front-to-back probe position (see Figure 17).
• Using the knurled nut on the left side and the +1 to -1 markers on the top front of the Ion
Max source housing, adjust the side-to-side probe position (see Figure 17).
• Using the A, B, C, and D markers on the probe as a guide, adjust the probe depth (see
Figure 15 on page 15).
Thermo Scientific
HESI-II Probe User Guide
17
3
Removing and Installing the HESI-II Probe
Adjusting the Probe Position
Figure 17. Top view of the Ion Max source housing with a HESI-II probe
Side-to-side
A
S
Knurled nut
Front-to-back
CAUTION - HOT SURFACE
1
1
+
0
0 1
Side-to-side
position set to 0
Micrometer
set to 1.75
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HESI-II Probe User Guide
Thermo Scientific
4
Maintenance
The HESI-II probe requires minimum maintenance. If the metal needle sample tube is
plugged, replace the needle insert. Replacing the needle insert requires a partial disassembly of
the probe.
Tip For best results, flush the HESI-II probe at the end of each working day, using a
50:50 LCMS-grade methanol/distilled water solution from the LC pump through the
HESI-II probe. To order LCMS grade solvents from Thermo Fisher Scientific, go to
www.FisherLCMS.com
IMPORTANT For best results, avoid operating the HESI-II probe at elevated temperatures
without solvent flow. Allowing the HESI-II probe to run dry at elevated temperatures can
cause blockage of the replaceable metal needle.
IMPORTANT For best results, wear clean gloves when you handle HESI-II probe
components.
CAUTION AVOID BURNS. At operating temperatures, the vaporizer can severely burn
you. The vaporizer typically operates between 350 and 450 °C. Always allow the heated
vaporizer to cool to room temperature (for approximately 20 min) before you remove or
touch the HESI-II probe.
Contents
• Flushing the Sample Transfer Line, Sample Tube, and HESI-II Probe
• Replacing the Needle Insert
Flushing the Sample Transfer Line, Sample Tube, and HESI-II Probe
For best results, flush the sample transfer line, sample tube, and HESI-II probe for 15 minutes
at the end of each working day (or more often if you suspect they are contaminated). Use a
50:50 methanol/distilled water solution from the LC system through the API source.
After 15 minutes, turn off the flow of liquid from the LC to the API source, but keep the API
source on (including the sheath gas and auxiliary gas) for an additional 5 minutes. Refer to the
daily operations chapter in the hardware manual for your mass spectrometer.
Thermo Scientific
HESI-II Probe User Guide
19
4
Maintenance
Replacing the Needle Insert
Replacing the Needle Insert
If the metal needle is plugged, you can replace the needle insert. The following procedure
describes how to replace the needle insert.
The needle insert is assembled at the factory and consists of an adjustable union, a needle
guide fitting, a ferrule, an O-ring, an ESI needle, and a metal needle (see Figure 18). The
ferrule is swaged onto the ESI needle. Factory adjusted, the metal needle protrudes 0.5 mm
from the end of the ESI needle (see Figure 19).
IMPORTANT Because the protrusion of the metal needle from the ESI needle is
factory-adjusted, do not disassemble the needle insert.
Figure 18. Exploded view of the needle insert
Metal needle
ESI needle
Needle guide fitting
and ferrule
Adjustable
union
Figure 19. Metal needle insert assembly with an enlarged view of the stainless steel needle tip
ESI needle
Metal
needle
20
HESI-II Probe User Guide
0.5 mm
Thermo Scientific
4 Maintenance
Replacing the Needle Insert
To support flow rates from 1 to 2000 μL/min, Thermo Fisher Scientific provides two needle
inserts for the HESI-II probe. The difference between the two inserts is the size of the metal
needle and supporting ferrule (Table 3).
Table 3. Needle inserts
Description
Metal needle
Ferrule
Flow rate range
32-gauge needle insert,
HESI-II probe
0.004 in. ID
0.009 in. OD
0.4 mm thru-hole
5 to 2000 μL/min
34-gauge needle insert,
HESI-II probe
0.003 in. ID
0.007 in. OD
0.2 mm thru-hole
1 to 10 μL/min
Replacing the needle insert requires a 3 mm (7/64 in.) hex wrench or ball driver.
Y To replace the needle insert
1. Remove the HESI-II probe from the Ion Max API source (see “Removing the HESI-II
Probe” on page 9).
2. Unscrew the fingertight fitting from the sample inlet port (see Figure 16 on page 16).
3. Remove the needle insert from the probe as follows (see Figure 20):
a. Using a 3 mm (7/64 in.) hex wrench or ball driver, remove the two M4×35 mm
length, socket head cap screws.
b. Pull the end cover off of the probe.
c. Unscrew the needle insert, and then pull it out of the probe body.
Figure 20. Exploded view of the HESI-II probe
Metal needle insert
End
cover
M4 ×35 mm length
socket head cap
screws (2×)
Thermo Scientific
Probe body
HESI-II Probe User Guide
21
4
Maintenance
Replacing the Needle Insert
4. Insert a new needle insert into the probe body.
5. Hand tighten the adjustable union fitting (see Figure 18 on page 20) until the needle
insert tip protrudes from the probe nozzle by 1.5 mm (see Figure 21).
Figure 21. Enlarged view of the probe nozzle
ESI needle
1.5 mm
Metal
needle
6. Position the end cover on the probe body.
7. Insert the two M4×35 mm length, socket head cap screws into the end cover, and then
tighten them with a 3 mm (7/64 in.) hex wrench or ball driver.
8. Reinstall the HESI-II probe (see “Installing the HESI-II Probe” on page 13).
22
HESI-II Probe User Guide
Thermo Scientific
5
Replaceable Parts
Use the following part numbers when you need to order the replaceable and consumable parts
for the HESI-II probe.
You can order these parts directly from Thermo Fisher Scientific.
HESI-II Probe Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . OPTON 20037
High-flow needle insert assembly . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . OPTON-53010
Low-flow needle insert assembly. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . OPTON-53011
The HESI-II Probe Kit contains the HESI-II probe with a factory-installed high-flow
needle insert. The high-flow metal needle insert is suitable for flow rates equal to or greater
than 5 μL/min. For low-flow rate applications, order the low-flow needle insert assembly. For
more information on the needle inserts, see Table 3 on page 21.
You can order these parts from Upchurch Scientific; however, these part numbers are subject
to change.
Fitting, fingertight, one-piece, for 1/16 in. OD high pressure tubing . . . . . . . . . . . . . F-120
Fitting, fingertight with one wing, two-piece,
red PEEK, for 1/16 in. OD high pressure tubing . . . . . . . . . . . . . . . . . . . . . . . . . F-200
Union, stainless steel, for 1/16 in. OD high pressure tubing . . . . . . . . . . . . . . . . . . . . U-435
Thermo Scientific
HESI-II Probe User Guide
23
I
Index
A
auxiliary gas flow
allowable range 4
plumbing diagram 8
B
blockage in the metal needle, frequent 19
C
CAUTIONS
avoiding an electric shock viii
avoiding burns, HESI-II probe vaporizer 19
cleaning procedure 19
compliance, WEEE iii
H-ESI
process, discussed 1
rules for good electrospray 4
HESI-II probe
figure 13
flushing 19
functional description 5
installing 13
photograph 1
removing 9
replaceable parts 23
vaporizer, avoiding burns (CAUTION) 19
L
locking ring, 8 kV cable 10
D
M
depth markers on probe body 15
E
maintenance
flushing the sample tube and transfer line 19
replacing the metal needle insert 20
micrometer, Ion Max 18
electrospray, rules for good electrospray 4
ESI needle, voltages 7
N
F
figures
auxiliary gas plumbing 8
ESI needle and metal needle protrusion 6
H-ESI process 3
HESI-II probe photograph 1
removable components of the HESI-II probe 5
sheath gas plumbing 7
front-to-back probe position 17
H
heated auxiliary gas, description 7
Thermo Scientific
needle insert, flow rate range 21
P
probe position, adjusting 17
R
red alignment markers 12
replaceable parts 23
S
safety precautions for handling the HESI-II probe viii
sheath gas plumbing diagram 7
side-to-side probe position 17
HESI-II Probe User Guide
25
Index: V
V
voltages, ESI needle 7
W
WEEE compliance iii
26
HESI-II Probe User Guide
Thermo Scientific
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