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Functional Description

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HESI-II Probe User Guide

Thermo Scientific

Figure 6.

Auxiliary gas plumbing

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”

Auxiliary gas fitting (green)
in gas inlet (A)

ESI needle

(outer needle)

Auxiliary gas

Metal needle
(inner needle)

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17
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Summary of Contents for HESI-II Probe

Page 1: ...HESI II Probe User Guide 70005 97000 Revision C April 2009 ...

Page 2: ...ion of Thermo Fisher Scientific Inc The contents of this document are subject to change without notice All technical information in this document is for reference purposes only System configurations and specifications in this document supersede all previous information received by the purchaser Thermo Fisher Scientific Inc makes no representations that this document is complete accurate or error f...

Page 3: ... Directives and the recyclers in your country WEEE Konformität Dieses Produkt muss die EU Waste Electrical Electronic Equipment WEEE Richtlinie 2002 96 EC erfüllen Das Produkt ist durch folgendes Symbol gekennzeichnet Thermo Fisher Scientific hat Vereinbarungen mit Verwertungs Entsorgungsfirmen in allen EU Mitgliedsstaaten getroffen damit dieses Produkt durch diese Firmen wiederverwertet oder ents...

Page 4: ...ieurs compagnies de recyclage dans chaque état membre de l union européenne et ce produit devrait être collecté ou recyclé par celles ci Davantage d informations sur la conformité de Thermo Fisher Scientific à ces directives les recycleurs dans votre pays et les informations sur les produits Thermo Fisher Scientific qui peuvent aider la détection des substances sujettes à la directive RoHS sont di...

Page 5: ...ntroduction 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 Contents ...

Page 6: ......

Page 7: ...ide 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 o...

Page 8: ...Standby mode before you tighten the fitting to eliminate the leak 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 CAUTION AVOID ELECTRIC SHOCK Do not tighten the probe sample inlet fitting to el...

Page 9: ...ssupport 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 techpubs lcms thermofisher com Phone 800 532 4752 Fax 561 688 8736 E mail us techsupport analyze therm...

Page 10: ......

Page 11: ...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 v...

Page 12: ...ist 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 criti...

Page 13: ...arge 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 ...

Page 14: ...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 a...

Page 15: ...ecured 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 prob...

Page 16: ...nt 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 Sheath gas inlet S Aux gas inlet A High voltage socket for the 8 kV cable connector Sample inlet Grounding union holder...

Page 17: ...lizes 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 ...

Page 18: ...x 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 Auxiliary gas fitting green in gas inlet A ESI needle outer nee...

Page 19: ...nected 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 Note To install the APCI probe refer to the Ion Max and Ion Max S API Source ...

Page 20: ...e 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 Sheath gas line Vaporizer cable To LC outlet 8 kV cable Auxiliary gas line Connection between grounding union and sample inlet Probe locking knob Locking ring High vol...

Page 21: ...I probe 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 A S Vaporizer cable connected to the probe 8 kV cable disconnected Probe vaporizer cable socket A S Sheath g...

Page 22: ...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 i...

Page 23: ...ssary 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 Note To remove an APCI or ESI probe refer to the Ion Max and Ion Max S API Source Hardware Manual Vaporizer cable connector socket Guide pin Sheath gas inlet Auxiliary gas inlet ...

Page 24: ... 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 Probe locking knob Probe locking ring Probe port Probe in...

Page 25: ...oser 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 va...

Page 26: ...ample 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 Tip Before analyzing samples change the ion source mode in TSQ EZ Tune by choosing Setup Change Ion Source HESI A S Aux...

Page 27: ...your application 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 d...

Page 28: ... Probe Position 18 HESI II Probe User Guide Thermo Scientific Figure 17 Top view of the Ion Max source housing with a HESI II probe A S CAUTION HOT SURFACE 1 1 0 1 0 Micrometer set to 1 75 Side to side position set to 0 Side to side Knurled nut Front to back ...

Page 29: ...in the hardware manual for your mass spectrometer 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 wi...

Page 30: ... 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 Figure 18 Exploded view of the needle insert Figure 19 Metal needle insert assembly with an enlarged view of the stainless steel needle tip IMPORTANT Because the protrusion of the metal needle from the ESI n...

Page 31: ...ngertight 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 Table 3 Ne...

Page 32: ... the needle insert tip protrudes from the probe nozzle by 1 5 mm see Figure 21 Figure 21 Enlarged view of the probe nozzle 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 ESI needle Metal ...

Page 33: ...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 fingertig...

Page 34: ......

Page 35: ...nts of the HESI II probe 5 sheath gas plumbing 7 front to back probe position 17 H heated auxiliary gas description 7 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 M maintenance flushing the sample tube an...

Page 36: ...26 HESI II Probe User Guide Thermo Scientific Index V V voltages ESI needle 7 W WEEE compliance iii ...

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