This guide describes how to install the NSI-1 dynamic nanospray probe (see Figure 1) onto a TSQ™ Series or LTQ™ Series mass spectrometer and provides general operation guidelines.

Figure 1. NSI-1 dynamic nanospray probe

The NSI source for the dynamic nanospray mode consists of the nanospray translation flange with an attached grounding union holder, the dynamic nanospray probe (NSI-1), and the nanospray base (NSI-1).

Dynamic Nanospray Probe (NSI-1) Installation Guide

Note Thermo Fisher Scientific has discontinued the NSI-2 dynamic nanospray probe.

Contents

• Required Parts

• Ion Max Adapter Installation

• Nanospray Translation Flange Assembly

• Emitter Installation

• Probe Installation

• NSI Source Installation

• Fused-Silica Transfer Line Installation

• Emitter Alignment

• Operation Guidelines

• Trademarks

Emitter

Snap-in nut

MicroTee mounting block

High-voltage pin lead (A) to liquid junction

High-voltage pin lead (B) to coated tip

© 2010 Thermo Fisher Scientific Inc.All rights reserved.

Revision A 97055-97100

RequiredParts

To install the NSI-1 dynamic nanospray probe, you must have the following parts: OPTON-20050 and OPTON-97017.

Table 1 lists the parts in OPTON-20050. To install the NSI-1 dynamic probe, you need the Ion Max™ adapter and the nanospray translation flange.

Table 2 lists the parts in OPTON-97017. This kit contains the NSI-1 probe with the nanospray base and cover, the grounding union holder, and the grounding union.

Ion MaxAdapter

Installation

The NSI-1 source mounts onto the Ion Max adapter.

To install the adapter hardware for the nanospray mode1. Place the mass spectrometer in standby mode. 2. Unplug the high-voltage connector from the ion source.

Table 1. Nanospray Interface Ion Max Kit OPTON-20050

Part description

Ion Max adapter

Nanospray translation flange

NSI-2 base (for the static nanospray probe)

1 in. deep adapter ring (not required)

2 in. deep adapter ring (not required)

Table 2. Dynamic nanospray probe with mount (NSI-1) and accessory kit (OPTON 97017)

Description Part number

Nanospray probe assembly (NSI-1) 97044-60280

Grounding union holder N/A

Socket head cap screws, 6-32 thread, 1/4 in. length, stainless steel (used to secure the grounding union holder to the nanospray translation flange)

00419-63204

Nanospray Accessory Kit 97044-62166

High-voltage emitter assembly 97044-60290S

MicroFerrule fitting, for 0.025 in. OD tubing, black PEEK, (Upchurch P/N F-172) 00101-18212

MicroFerrule tubing sleeve, 0.015 in. ID × 0.025 in. OD × 1.0 in. length, green PEEK (Upchurch P/N F-185x)

00101-50060

PicoTip™, glass (included in kit, but not used with the dynamic probe) N/A

Adaptor nut, snap-in N/A

Socket head cap screw, 2-56 thread, 3/16 in. length, stainless steel 00405-63202

Emitter clip N/A

Contact, PicoTip N/A

MicroTight™ zero dead volume union, 0.010 in. thru hole, 10-32 flat bottom ports 00101-18182

Fused silica tubing, 50 μm ID × 360 μm OD, 1 m length 00106-10503

Dynamic nanospray emitters, 30 μm ID tip, metal coated, 12.0 cm length, box of 2 00301-57008

Ferrule, 0.016 in. ID, natural PEEK (for use with fused-silica transfer line) 00101-18120

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3. Install the Ion Max adapter on your mass spectrometer as follows:a. If you have not already done so, remove the Ion Max ion source as described in the Ion Max and Ion Max-S

API Source Hardware Manual. b. Carefully align the two guide pin holes on the back of the Ion Max adapter (see Figure 2) with the guide pins

on the mass spectrometer (see Figure 3). Figure 2. Ion Max adapter, showing the guide pin holes and the alignment tab

Figure 3. Mass spectrometer, showing ion source guide pins

c. Ensure that the Ion Max adapter locking levers are open (pointing away from the mass spectrometer), and gently push the adapter onto the mass spectrometer.

d. Rotate the locking levers 90 degrees to lock the adapter onto the mass spectrometer.

Ion source guide pin holes (back view)

Alignment tab(front view)

Ion sourceguide pins

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NanosprayTranslation

FlangeAssembly

The original NSI-1 nanospray translation flange assembly consisted of the nanospray translation flange, the NSI-1 base, and the grounding union holder (see Figure 4). This assembly is no longer preassembled at the factory. OPTON-20050 contains the nanospray translation flange and OPTON-97017 contains the NSI-1 base and the grounding union holder.Figure 4. NSI-1 nanospray translation flange assembly parts

To assemble the nanospray translation flange assembly1. Slide the NSI-1 base onto the nanospray translation flange (see Figure 5).

Figure 5. Installing the NSI-1 base

Tools needed

• 7/64 in. L-hex wrench

• #1 Phillips screw driver

Note In the following procedure, the photographs show the conversion of an NSI-2 source to an NSI-1 source.

Nanospray translation flange

Grounding union holder

NSI-1 base with probe

Nanospray translation flange

NSI-1 base

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2. Using a #2 Phillips screwdriver, tighten the screw that secures the NSI-1 base to the flange (see Figure 6).Figure 6. Tightening the Phillips head screw

3. Using two socket head cap screws (6-32 thread, 1/4 in. length) and an L-hex wrench, secure the NSI-1 grounding union holder to the nanospray translation flange (see Figure 7). Figure 7. Installing the grounding union holder

Nanospray translation flange

Screw that secures the NSI-1 base to the nanospray translation flange

Grounding union holder

L-hex wrench inserted into the head of one of the screws

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EmitterInstallation

Figure 8 shows an exploded view of the NSI-1 dynamic nanospray probe assembly.

Figure 8. NSI-1 dynamic nanospray probe, fused silica emitter, and fused silica liquid transfer line (exploded view)

To install a fused-silica emitter1. Using a felt tip marker, mark the emitter 10.5 cm from its tip. 2. Slide a green MicroTight sleeve (P/N 00101-50060), followed by the snap-in nut (threaded end first), over the

blunt end of the emitter.3. Slide a MicroFerrule (P/N 00101-18212) over the green MicroTight sleeve.4. Cut the emitter at the mark you made in step 1. 5. Rinse the cut end of the emitter with an LC-MS grade solvent (such as 100% methanol or 100% acetonitrile).

Then slide the sleeve back until it is flush with the blunt end of the emitter.6. Insert the blunt end of the sleeved emitter into the emitter port of the MicroTee fitting (see Figure 9).

CAUTION Avoid touching the emitter tip, as it is sharp enough to puncture the skin.

IMPORTANT Avoid touching any hard surface with the emitter tip, as it is easily damaged or broken.

Tools needed

• 5/64 in. Allen key

• Extra-sharp, stainless steel tweezers

Snap-in tip mount Fused-silica emitter Snap-in nut

MicroTee mounting block and in-union high voltage contact

Fused-silica liquid transfer line

Fingertight II nut, red (P/N 00101-18195)and MicroFerrule (P/N 00101-18212)

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Figure 9. MicroTee fitting, sleeved emitter, MicroFerrule, and snap-in nut

7. With the sleeved emitter held firmly against the bottom of the port, screw the snap-in nut onto the MicroTee until it is hand-tight.

8. Using a 5/64 in. L-hex wrench, loosen the 2-56 socket head screw (see Figure 10) at the end of the snap-in tip mount by half a turn.Figure 10. Socket head screw and clip at the end of the snap-in mount

9. Holding the clip up with extra-sharp tweezers, slide the emitter capillary under the clip and into the groove of the snap-in mount.

10. Ensure that the emitter tip projects about 4 mm beyond the end of the snap-in mount. 11. Hand tighten the emitter clip socket screw. Do not over tighten the screw.

Emitter port

MicroFerrule(Upchurch P/N F-172)

Snap-in end of snap-in nut

MicroTight sleeve

Emitter

Internally threaded end of the snap-in nut

Socket head screw

Clip

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ProbeInstallation

To install the NSI-1 probe onto the NSI -1 base1. Slide the NSI-1 dynamic probe assembly into the NSI -1 base until it snaps into place (see Figure 11).

Figure 11. NSI-1 dynamic nanospray probe seated in NSI-1 base

2. Depending on whether you are applying high voltage to a liquid junction or an emitter tip, do the following:

• To apply voltage to a liquid junction, connect the high voltage socket lead on the NSI-1 base to the high voltage pin lead (A) coming from the MicroTee high voltage port.

• To apply voltage directly to a coated emitter tip, connect the high voltage socket lead on the NSI-1 base to the high voltage pin lead (B) coming from the NSI-1 dynamic probe.

3. Slide the probe cover (see Figure 12) over the NSI-1 base. Figure 12. Probe cover (front view)

4. To secure the safety cover to the NSI-1 base, screw the internally-threaded thumbscrew onto the externally-threaded nut protruding from the back of the NSI-1 base (see Figure 13 and Figure 15).

High voltage pin lead (B)

High voltagesocket lead

High voltage pin lead (A)

NSI -1 base

Probe cover

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Figure 13. NSI-1 base with safety cover (back view)

NSI SourceInstallation

To install the NSI source1. Turn the Z-position knob counterclockwise until the XYZ stage is fully retracted (see Figure 14).

Figure 14. NSI source

2. Install the NSI source onto the Ion Max adapter as follows:a. Align the alignment slot on the nanospray translation flange (see Figure 15) with the alignment tab on the

Ion Max adapter (see Figure 2 on page 3), and then slide the flange onto the adapter.

Safety cover

Externally-threaded nut

Internally-threadedThumbscrew

Z position adjustment knob

Flange retainer bolts

High voltage connector

XYZ stage

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Figure 15. Alignment slot on the bottom of the nanospray translation flange

b. To secure the flange to the adapter, hand tighten the two retainer bolts (see Figure 14 on page 9). c. Connect the mass spectrometer’s high voltage cable to the high voltage cable connector on the NSI-1 base.

Secure the cable by turning the locking ring clockwise.

Fused-SilicaTransfer Line

Installation

To connect a fused-silica transfer line from the grounding union to the probe1. Using a fused-silica tubing cutter, cut a 12 in. (30 cm) length of fused-silica capillary tubing (50 μm ID). 2. Slide a MicroTight nut, a green MicroTight tubing sleeve (P/N 00101-50060) and a MicroFerrule

(P/N 00101-18212) onto one end of the fused-silica transfer line. Adjust the sleeve so that it is flush with the end of the fused-silica transfer line.

3. Insert the sleeved fused-silica transfer line into the transfer line port of the MicroTee (see Figure 16).Figure 16. Fused-silica connection to the MicroTee

4. With the sleeved tubing and MicroFerrule held firmly against the bottom of the MicroTee port, tighten the MicroTight nut.

Alignment slot

Thumbscrew

Fused-silica transfer line

MicroTight nut

MicroFerrule

MicroTightsleeve

MicroTee

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5. Using a Fingertight II nut (Upchurch P/N F-200) and a 0.016 in. ID ferrule (P/N 00101-18120) or a ferrule designed for 360 μm OD tubing (Upchurch P/N F-151), connect the other end of the fused-silica transfer line to the grounding union (see Figure 17).Figure 17. Fused-silica connection to the MicroTee

EmitterAlignment

Align the emitter tip with the center of the ion transfer tube.

To align the emitter tip1. Use the X and Y adjustment knobs on the XYZ stage to align the emitter tip with the center of the ion transfer

tube.

Position the emitter no more than 1 mm from the center of the ion transfer tube to ensure spray quality.2. Using the Z adjustment knob, position the emitter approximately 2 mm away from the ion transfer tube.

OperationGuidelines

Table 3 through Table 6 display the NSI method parameters for operation on an LTQ Series or a TSQ Series mass spectrometer. As you progress to smaller ID columns (micro-capillary to nano-capillary), the suggested flow rate, emitter tip ID, and capillary temperature decrease.

Fused-silica transfer line connection to the grounding union

IMPORTANT If you are transferring a nanospray method developed on an LTQ or LTQ XL™ mass spectrometer to an LTQ Velos™ mass spectrometer or from a TSQ Quantum™ mass spectrometer to a TSQ Vantage™ mass spectrometer, raise the capillary temperature (for the ion transfer tube heater) by 50 °C. The minimum capillary temperature for the LTQ Velos or TSQ Vantage mass spectrometer in the NSI mode is 250 °C.

IMPORTANT When you are developing a nanospray method for the LTQ Velos or TSQ Vantage mass spectrometer, use a minimum capillary temperature of 250 °C. Setting the capillary temperature below 250 °C can result in contamination of the ion source interface and more frequent maintenance of the ion transfer tube.

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Trademarks TSQ, TSQ Vantage, LTQ Velos, and Ion Max are trademarks; and LTQ and TSQ Quantum are registered trademarks of Thermo Fisher Scientific Inc. in the United States.

MicroTight and Upchurch Scientific are registered trademarks of IDEX Health & Science LLC in the United States.

PicoTip is a registered trademark of New Objective, Inc. in the United States.

Note The values listed in Table 3 through Table 6 can vary significantly from emitter to emitter. Refer to these values as guidelines, not specifications.

The tip ID sizes are based on PicoTips information provided by New Objective, Inc., Cambridge, Massachusetts. Go to the New Objective Web site (http://www.newobjective.com)or its catalogs for more information on the various tip sizes and types.

The emitter position refers to the distance of the emitter tip from the surface of the ion transfer tube.

Table 3. LC/NSI/MS parameters for an LTQ or LTQ XL mass spectrometer

Column size Flow rate Tip ID Emitter position Capillary temperature

50 μm <100 to 300 nL/min 5 to 15 μm approx. 1 mm 100 to 180 °C

75 μm 150 to 800 nL/min 10 to 20 μm 1 to 2 mm 130 to 200 °C

180 μm 0.3 to 1.0 μL/min 20 to 30 μm 2 to 3 mm 150 to 250 °C

320 μm 1 to 5 μL/min 30 to 75 μm 3 to 5 mm 150 to 250 °C

Table 4. LC/NSI/MS parameters for an LTQ Velos mass spectrometer

Column size Flow rate Tip ID Emitter position Capillary temperature

50 μm <100 to 300 nL/min 5 to 15 μm approx. 1 mm minimum 250 °C

75 μm 150 to 800 nL/min 10 to 20 μm 1 to 2 mm minimum 250 °C

180 μm 0.3 to 1.0 μL/min 20 to 30 μm 2 to 3 mm 250 ° to 300 °C

320 μm 1 to 5 μL/min 30 to 75 μm 3 to 5 mm 250 ° to 300 °C

Table 5. LC/NSI/MS parameters for the TSQ Quantum family of mass spectrometers

Column size Flow rate Tip ID Emitter position Capillary temperature

50 μm <100 to 300 nL/min 5 to 15 μm approx. 1 mm 225 to 275 °C

75 μm 150 to 800 nL/min 10 to 20 μm 1 to 2 mm 225 to 275 °C

180 μm 0.3 to 1.0 μL/min 20 to 30 μm 2 to 3 mm 225 to 275 °C

320 μm 1 to 5 μL/min 30 to 75 μm 3 to 5 mm 225 to 300 °C

Table 6. LC/NSI/MS parameters for the TSQ Vantage mass spectrometer

Column size Flow rate Tip ID Emitter position Capillary temperature

50 μm <100 to 300 nL/min 5 to 15 μm approx. 1 mm 250 to 325 °C

75 μm 150 to 800 nL/min 10 to 20 μm 1 to 2 mm 250 to 325 °C

180 μm 0.3 to 1.0 μL/min 20 to 30 μm 2 to 3 mm 250 to 325 °C

320 μm 1 to 5 μL/min 30 to 75 μm 3 to 5 mm 250 to 350 °C

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