Dynamic Nanospray Probe (NSI-1) Installation Guide
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1 Dynamic Nanospray Probe (NSI-1) Installation Guide 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. 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 Figure 1. NSI-1 dynamic nanospray probe MicroTee mounting block Snap-in nut Emitter High-voltage pin lead (A) to liquid junction High-voltage pin lead (B) to coated tip 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). Revision A Thermo Fisher Scientific Inc. All rights reserved.
2 Required Parts To install the NSI-1 dynamic nanospray probe, you must have the following parts: OPTON and OPTON Table 1 lists the parts in OPTON To install the NSI-1 dynamic probe, you need the Ion Max adapter and the nanospray translation flange. Table 1. Nanospray Interface Ion Max Kit OPTON 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 lists the parts in OPTON This kit contains the NSI-1 probe with the nanospray base and cover, the grounding union holder, and the grounding union. Table 2. Dynamic nanospray probe with mount (NSI-1) and accessory kit (OPTON 97017) Description Part number Nanospray probe assembly (NSI-1) 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) Nanospray Accessory Kit High-voltage emitter assembly S MicroFerrule fitting, for in. OD tubing, black PEEK, (Upchurch P/N F-172) MicroFerrule tubing sleeve, in. ID in. OD 1.0 in. length, green PEEK (Upchurch P/N F-185x) 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 Emitter clip N/A Contact, PicoTip N/A MicroTight zero dead volume union, in. thru hole, flat bottom ports Fused silica tubing, 50 μm ID 360 μm OD, 1 m length Dynamic nanospray emitters, 30 μm ID tip, metal coated, 12.0 cm length, box of Ferrule, in. ID, natural PEEK (for use with fused-silica transfer line) Ion Max Adapter Installation The NSI-1 source mounts onto the Ion Max adapter. To install the adapter hardware for the nanospray mode 1. Place the mass spectrometer in standby mode. 2. Unplug the high-voltage connector from the ion source. 2
3 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 Ion source guide pin holes (back view) Figure 3. Mass spectrometer, showing ion source guide pins Alignment tab (front view) 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. 3
4 Nanospray Translation Flange Assembly 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 contains the nanospray translation flange and OPTON contains the NSI-1 base and the grounding union holder. Figure 4. NSI-1 nanospray translation flange assembly parts NSI-1 base with probe Grounding union holder Nanospray translation flange 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. To assemble the nanospray translation flange assembly 1. Slide the NSI-1 base onto the nanospray translation flange (see Figure 5). Figure 5. Installing the NSI-1 base Nanospray translation flange NSI-1 base 4
5 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 Nanospray translation flange Screw that secures the NSI-1 base to the nanospray translation flange 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 L-hex wrench inserted into the head of one of the screws Grounding union holder 5
6 Emitter Installation Figure 8 shows an exploded view of the NSI-1 dynamic nanospray probe assembly. 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. Figure 8. NSI-1 dynamic nanospray probe, fused silica emitter, and fused silica liquid transfer line (exploded view) Snap-in tip mount Fused-silica emitter Snap-in nut Fused-silica liquid transfer line MicroTee mounting block and in-union high voltage contact Fingertight II nut, red (P/N ) and MicroFerrule (P/N ) Tools needed 5/64 in. Allen key Extra-sharp, stainless steel tweezers To install a fused-silica emitter 1. Using a felt tip marker, mark the emitter 10.5 cm from its tip. 2. Slide a green MicroTight sleeve (P/N ), followed by the snap-in nut (threaded end first), over the blunt end of the emitter. 3. Slide a MicroFerrule (P/N ) over the green MicroTight sleeve. 4. Cut the emitter at the mark you made in step 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). 6
7 Figure 9. MicroTee fitting, sleeved emitter, MicroFerrule, and snap-in nut Internally threaded end of the snap-in nut MicroFerrule (Upchurch P/N F-172) Emitter port Emitter Snap-in end of snap-in nut MicroTight sleeve 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 Clip Socket head screw 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. 7
8 Probe Installation To install the NSI-1 probe onto the NSI -1 base 1. 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 NSI -1 base High voltage pin lead (A) High voltage pin lead (B) High voltage socket lead 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) Probe cover 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). 8
9 Figure 13. NSI-1 base with safety cover (back view) Safety cover Internally-threaded Thumbscrew Externally-threaded nut NSI Source Installation To install the NSI source 1. Turn the Z-position knob counterclockwise until the XYZ stage is fully retracted (see Figure 14). Figure 14. NSI source Flange retainer bolts High voltage connector Z position adjustment knob XYZ stage 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. 9
10 Figure 15. Alignment slot on the bottom of the nanospray translation flange Thumbscrew Alignment slot 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-Silica Transfer Line Installation To connect a fused-silica transfer line from the grounding union to the probe 1. 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 ) and a MicroFerrule (P/N ) 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 MicroTight sleeve Fused-silica transfer line MicroTee MicroFerrule MicroTight nut 4. With the sleeved tubing and MicroFerrule held firmly against the bottom of the MicroTee port, tighten the MicroTight nut. 10
11 5. Using a Fingertight II nut (Upchurch P/N F-200) and a in. ID ferrule (P/N ) 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 Fused-silica transfer line connection to the grounding union Emitter Alignment Operation Guidelines Align the emitter tip with the center of the ion transfer tube. To align the emitter tip 1. 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. 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. 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. 11
12 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 ( 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 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. 12
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