HESI Probe User Guide Version C
Contents
1. 2009 Thermo Fisher Scientific Inc All rights reserved The following are registered trademarks in the United States and other countries Teflon and Tefzel are registered trademarks of E I du Pont de Nemours amp Co KEL F is a registered trademark of 3M Corporation PEEK is a trademark of Victrex plc Ton Max is a trademark of Thermo Fisher Scientific Inc in the United States Xcalibur is a registered trademark of Thermo Fisher Scientific Inc in the United States All other trademarks are the property of Thermo Fisher Scientific Inc and its subsidiaries Thermo Fisher Scientific Inc provides this document to its customers with a product purchase to use in the product operation This document is copyright protected and any reproduction of the whole or any part of this document is strictly prohibited except with the written authorization 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 free and assumes no responsibility and will not be liable for any errors omissions damage or loss that might result from any use of this document even if the information in the document is followed pro2. 3000 2500 200 350 250 to 300 35 30 3000 2500 500 380 300 to 400 60 50 3000 2500 1000 400 350 to 450 75 60 3000 2500 x Always optimize the tube lens voltage whenever you change the temperature of the ion transfer tube aK Compound dependent KK a ax m y For an LTQ Series mass spectrometer controlled from LTQ 2 50 or lower the minimum auxiliary gas flow setting is 5 sek _ Negative ion mode 4 H ESI Probe User Guide Thermo Scientific Functional Description Thermo Scientific The H ESI probe shown in Figure 3 produces charged aerosol droplets that contain sample ions The H ESI probe accommodates liquid flows of 1 pL min to 1 mL min without splitting The H ESI probe includes a fused silica sample tube needle nozzle manifold vaporizer and heat exchanger Sample and solvent enter the H ESI probe through the sample tube The sample tube is a short section of 0 1 mm ID fused silica tubing that extends through the H ESI probe and into the H ESI needle to near the end of the H ESI needle A large negative or positive voltage is applied to the H ESI needle typically 3 to 5 kV which sprays the sample solution into a fine mist of charged droplets The H ESI nozzle directs the flow of sheath gas and auxiliary gas at the droplets The H ESI manifold houses the H ESI nozzle and needle and includes the sheath gas and auxiliary gas plumbing The sheath gas plumbing delivers dry nitrogen gas to spray the sample s
3. G guide pin location figure 6 H heated auxiliary gas description 5 H ESI change ion source tip 12 process figure 2 process discussed 1 rules for optimizing 3 H ESI needle high voltage connector location figure 6 replacing 14 voltages 5 H ESI probe disassembling 14 exploded view figure 15 figure 1210 flushing 13 functional description 5 installed figure 8 12 installing 9 maintenance overview 13 PEEK safety sleeve 16 reassembling 16 removing 7 replaceable parts 19 sample tube description of 5 sample tube installing 16 H ESI sample tube trimming 14 H ESI vaporizer avoiding burns CAUTION 13 cable socket location figure 6 Ion Max figure 8 9 12 ion source interface figure 1 H ESI Probe User Guide 21 Index M 22 maintenance H ESI probe overview 13 needle seal replacing 14 needle replacing 14 sample transfer line flushing 13 sample tube flushing 13 sample tube installing 16 sample tube trimming 14 needle seal location figure 15 reinstalling 16 removing 14 needle H ESI location figure 15 reinstalling 16 removing 14 P part numbers for replaceable parts 19 PEEK safety sleeve CAUTION 16 installing 16 installing figure 17 replaceable parts for the H ESI probe 19 S safety precautions for handling the HESI II probe viii sample inlet location of figure 6 sample transfer line flushing 13 sample tube flushing 13 installing 16 installing figure
4. 18 repositioning note 18 trimming 14 sheath gas description 5 flow rates 5 sheath liquid description of 6 solvents ordering 19 T tuning frequency note 3 H ESI Probe User Guide V voltages applied to the H ESI needle 5 W WEEE compliance iii Thermo Scientific
5. ESI Probe When operating the mass spectrometer in the heated electrospray mode do not touch the heated surfaces of the H ESI 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 H ESI probe and the API source housing 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 viii H ESI Probe User Guide Thermo Scientific Preface Contacting Us There are several ways to contact Thermo Fisher Scientific for the information you need To contact Technical Support Phone 800 532 4752 Fax 561 688 8736 E mail us techsupport analyze thermofisher com Knowledge base www thermokb com Find software updates and utilitie
6. Sample Tube and PEEK Safety Sleeve 18 H ESI Probe User Guide 11 Slide a brown Fingertight fitting P N 00101 18081 and brown ferrule P N 00101 18119 wide end first over the PEEK safety sleeve 12 Connect the PEEK safety sleeve and the ferrule to the stainless steel ZDV fitting by tightening the brown Fingertight fitting Ensure that the Fingertight fitting is securely tightened around the PEEK safety sleeve otherwise the sample stream might enter between the sample tube and the PEEK safety sleeve Ensure the sample tube is held tightly in the grounded fitting by gently pulling the sample tube from the exit end of the H ESI needle 13 Use a fused silica cutting tool to cut the sample tube at the H ESI needle so that only 2 5 cm 1 in remains protruding from the exit end of the H ESI needle 14 From the sample inlet pull the PEEK safety sleeve backwards so that the exit end of the sample tube is recessed just inside the H ESI needle by approximately 0 5 mm between 0 and 1 mm See Figure 11 15 Tighten the red Fingertight fitting securely to hold the PEEK safety sleeve and sample tube in place Note The sample tube might move forward when you tighten the sample inlet fitting Ensure that the sample tube is retracted into the H ESI needle approximately 0 5 mm between 0 and 1 mm If necessary loosen the fitting and reposition the sample tube Figure 11 Installing the fused silica sample tube H ESI prob
7. 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 H ESI Probe User Guide 1 1 Introduction 2 H ESI Probe User Guide In H ESI ions are produced and analyzed as follows 1 2 6 The sample solution enters the H ESI needle which receives a high voltage The H ESI needle sprays the sample solution into a fine mist of droplets that are electrically charged at their surface The electrical charge density at the surface of the droplets increases as solvent evaporates from the droplets In H ESI heated auxiliary gas aids solvent evaporation 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 Electrostatic repulsion ejects sample ions from the very small highly charged droplets into the gas phase The sample ions enter the mass spectrometer and are analyzed Figure 2 shows the steps in the formation of ions from highly charged droplets Figure 2 H E
8. safety kit discussed below You could receive an electrical shock if the fused silica capillary tube breaks during H ESI operation Therefore for your safety and in compliance with international safety standards you must cover the fused silica capillary tube with the PEEK safety sleeve P N 00301 22806 and associated PEEK ferrules P N 00101 18119 provided in the Safety Sleeve Kit P N 70005 62015 before you operate the instrument Installation instructions P N 70005 97009 are included in the kit Operation of the instrument without the safety sleeve impairs the safety protection provided by the instrument and thus could lead to serious injury 1 Remove the H ESI probe from the Ion Max source by following the procedure in the topic Removing the H ESI Probe on page 7 Thermo Scientific 4 Maintenance Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve 2 Use a fused silica cutting tool to cut a 30 cm 12 in piece of 0 1 mm ID x 0 19 mm OD fused silica tubing sample tube P N 00106 10499 Ensure that you cut the ends of the fused silica tubing squarely 3 Insert the sample tube through the exit end of the H ESI needle and into the H ESI probe 4 Push the sample tube through the H ESI probe until approximately 3 5 cm 1 5 in is left protruding from the exit end of the H ESI needle The remaining length of sample tube should exit the H ESI probe sample inlet 5 Slide the 10 32 x 1 4 28 Kel F fitting a
9. two 6 32 x 1 4 in socket screws Remove the end cover from the H ESI manifold Thermo Scientific 4 Maintenance Replacing the H ES Needle and Needle Seal 6 Unscrew and remove the sheath liquid calibrant fitting 7 Remove the liquid junction union needle needle seal and needle sheath from the H ESI manifold as a unit See Figure 9 8 While grasping the liquid junction union with a 1 2 in open end wrench use a 3 8 in open end wrench to unscrew the needle sheath from the liquid junction union 9 Remove the H ESI needle and needle seal from the needle sheath 10 Remove the needle seal from the H ESI needle 11 Inspect the Teflon needle seal P N 97055 20271 If the needle seal is deformed replace it 12 Inspect the 26 gauge H ESI needle P N 97055 20273 If the H ESI needle is damaged replace it If you replace the needle you should also replace the needle seal 13 Reinstall the H ESI probe as described in the topic Installing the H ESI Probe on page 9 Figure 9 H ESI probe exploded view Needle seal H ESI I Liquid junction union j a needle High voltage connector receptacle a 6 32 x 1 5 in socket screws Needle sheath H ESI manifold 10 32 x 1 4 in HPLC adaptor fitting Es 10 32 x 1 4 in HPLC adaptor fitting Thermo Scientific H ESI Probe User Guide 15 4 Maintenance Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve 16 To
10. 8 kV cable H ESI vaporizer cable H ESI needle Sheath gas line Auxiliary gas line blue fitting green fitting Sample inlet red fitting PEEK safety sleeve Grounding union Probe locking knob The H ESI source is now properly installed on the mass spectrometer Tip Before you analyze samples with the H ESI source you must change to H ESI source mode in Quantum Tune Master by choosing Setup gt Change Ion Source gt HESI 12 H ESI Probe User Guide Thermo Scientific Maintenance The H ESI probe requires minimum maintenance If the fused silica sample tube is plugged or broken you must replace it Although you can trim or replace the sample tube without disassembling the H ESI probe to replace the H ESI needle or needle seal you must partially disassemble the H ESI probe Tip For best results flush the H ESI probe at the end of each working day using a 50 50 LCMS grade methanol distilled water solution from the LC through the H ESI probe Wear clean gloves when you handle H ESI probe components CAUTION AVOID BURNS At operating temperatures the H ESI vaporizer can severely burn you The H ESI 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 H ESI probe Contents e Flushing the Sample Transfer Line Sample Tube and H ESI Probe e Trimming the H ESI Sample Tube e Replacing th
11. SI process in the positive ion polarity mode H ESI probe H ESI needle cone lon transfer capillary D ion skimmer Charged Solvent Ejected droplet evaporates positive ions from droplet Thermo Scientific Thermo Scientific 1 Introduction You can use the H ESI mode 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 Use a positively charged needle to analyze positive ions and a negatively charged needle to analyze negative ions as the ejection of sample ions from droplets is facilitated if the ionic charge and surface charge of the droplet have the same polarity 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 Larg
12. Unlock the probe locking ring by turning the probe locking knob counterclockwise Carefully pull the probe straight back in the port in the housing until it meets with the slot in the ESI interlock block The guide pin on the probe manifold prevents you from twisting the probe until the pin is aligned with the slot in the ESI interlock block Once the probe is all the way back and aligned with the slot turn the probe 45 degrees counterclockwise to free the probe from the alignment notch Be careful not to break the fused silica sample tube or PEEK safety sleeve 10 Pull the probe straight out to remove it from the ion source housing 11 Store the H ESI probe in its original shipping container 8 H ESI Probe User Guide Thermo Scientific 3 Removing and Installing the H ESI Probe Installing the H ESI Probe Installing the H ESI Probe Note To remove an APCI or ESI probe refer to the Jon Max and Ion Max S API Source Hardware Manual To install the H ESI probe 1 Remove the H ESI probe from its storage container Inspect and clean it if necessary Note If your H ESI probe does not already have a sample tube fused silica capillary and safety sleeve attached you must follow the procedure for installing a sample tube and PEEK safety sleeve in the topic Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve on page 16 2 Ensure that the probe locking ring is opened to its widest position See Figu
13. ample inlet H ESI needle Grounding union holder Sheath liquid calibrant inlet To enhance desolvation or to improve the stability of the electrospray you can infuse sheath liquid directly from the syringe pump into the H ESI probe via the sheath liquid calibrant inlet labeled CAL You can also infuse internal calibrants into the H ESI probe The angle of the H ESI 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 refer to the Ion Max and Ion Max S API Source Hardware Manual 6 H ESI Probe User Guide Thermo Scientific Removing and Installing the H ESI Probe This chapter describes how to remove and install the H ESI probe from the Ion Max source housing You must remove the H ESI probe to change ionization modes and to perform maintenance on the probe You do not need to remove the Ion Max source housing to change probes Note To remove or install an APCI or ESI probe refer to the lon Max and Ion Max S API Source Hardware Manual Removing the H ESI Probe CAUTION AVOID BURNS At operating temperatures the H ESI vaporizer can severely burn you The H ESI vapo
14. and tune the parameters for your H ESI experiment H ESI Probe User Guide 3 1 Introduction Table 1 shows initial H ESI settings for different liquid flow rates 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 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 e For LTQ 2 5 0 or lower the allowable range for the auxiliary gas flow is 5 to 60 units e 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 H ESI settings for the H ESI probe Liquid Ion anster fubs H ESI vaporizer SUNS Auxiliarygasflow Spray voltage Wt capillary temperature C pressure arbitrary units V pL min temperature C psi 5 240 Off to 50 5 0
15. daptor P N 00101 18080 over the sample tube see Figure 10 and tighten the fitting onto the H ESI probe sample inlet Figure 10 Fused silica sample tube and safety sleeve assembly H ESI probe Fingertight fitting Grounding union P N 00101 18195 stainless steel ZVD fitti 10 32 x 1 4 28 fitting Fingertight fitting eng P N 00101 18080 P N 00101 18081 QUI pes NEN C Ber Sample tube fused silica capillary Ferrule Ferrule P N 00106 10499 P N 00101 18119 P N 00101 18119 PEEK safety sleeve P N 00301 22806 Grounding union holder 6 Slide the precut 25 cm 10 in 0 009 in ID x 0 024 in OD PEEK safety sleeve P N 00301 22806 over the sample tube 7 Slide the 0 027 in ID PEEK ferrule P N 00101 18119 narrow end first over the PEEK safety sleeve and to the 10 32 x 1 4 28 Kel F fitting adaptor 8 Slide the red Fingertight fitting P N 00101 18195 onto the PEEK safety sleeve and into the H ESI probe sample inlet labeled Sample Tighten the fitting slightly but not completely 9 Push the PEEK safety sleeve over the sample tube until it stops against the Teflon needle seal inside the H ESI probe 10 Pull the sample tube from the H ESI needle end until the sample tube is flush with the precut square end of the PEEK safety sleeve Thermo Scientific H ESI Probe User Guide 17 4 Maintenance Installing a New Fused Silica
16. e H ESI Needle and Needle Seal e Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve Flushing the Sample Transfer Line Sample Tube and H ESI Probe Thermo Scientific For best results flush the sample transfer line sample tube and H ESI 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 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 your mass spectrometer s hardware manual H ESI Probe User Guide 13 4 Maintenance Trimming the H ESI Sample Tube 14 Trimming the H ESI Sample Tube Operating your instrument with acetonitrile in the mobile phase can cause the polyimide coating on the fused silica sample tube to elongate If the polyimide coating has elongated past the end of the H ESI needle you must cut and reposition the end of the sample tube Le A WW N To cut and reposition the end of the sample tube approximately 0 5 mm between 0 and 1 mm inside the end of the H ESI needle Remove the H ESI probe from the Ion Max source by following the procedure in the topic Removing the H ESI Probe on page 7 Loosen the sample inlet fitting Gently pull back on the sample tube to free it from the fitt
17. e Sample tube fused silica capillary H ESI needle Sample tube Thermo Scientific Replaceable Parts Thermo Scientific This chapter contains part numbers for replaceable and consumable parts for the H ESI probe To ensure proper results in servicing the H ESI probe order only the parts listed or their equivalent This chapter contains part numbers for the following items Probe H ESL 5 2 5 5 be Oia eed Se wd cas Oa Rie dede 97055 60140 Needle metal 26 gauge H ESI 40402400000 ee a 97055 20273 T C 004 0 anne dt nate had ain dee anna ae anaes 97055 20271 Tubing fused silica 0 10 mm ID x 0 19 mmOD 00106 10499 Fitting grounding union ZV 1 4 28 sa 002 iivaraee is 00109 00304 Ferrule 0 008 in ID KEL E HPLC 00101 18075 Fitting plug 1 4 28 TEFZEL HPLC u 00101 18075 Ferrule 0 012 in ID KEL K HPLC 00101 18116 Salety Sleeve Kit en aens essen 70005 62015 Ferrule 0 027 in ID PEEK HPLC 00101 18119 Safety sleeve 0 009 in ID x 024 in OD x 10 in length natural PEEK 3 640050 04 ei ee alah due 00301 22806 Instructions safety sk son ben aden Re PROS RUES 70005 97009 Fitting Fingertight Upchutchts sous se sue iteiabuutehatuens 00101 18195 Fitting Fingertight HPLC 10 32 PEEK zus 00101 18081 Fitting adaptor 10 32 x 1 4 28 KEL F anne 00101 18080 For information on ordering LCMS grad
18. e droplets with high surface tension low volatility strong ion solvation low surface charge and high conductivity prevent good electrospray 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 e Keep salts out of the solvent system e Use organic aqueous solvent systems and volatile acids and bases e Optimize the pH of the solvent system Note You do not need to tune and calibrate the mass spectrometer as part of your daily routine Calibration parameters are instrument parameters that affect the mass accuracy and resolution Tune parameters are instrument parameters that affect the intensity of the ion signal You must tune and calibrate the mass spectrometer that is optimize the tune and calibration parameters approximately once a quarter Refer to the Automatic Tuning and Calibrating in the H ESI MS Mode chapter in your instrument s getting started manual for a procedure to tune and calibrate your mass spectrometer You must optimize the tune parameters or change the Tune Method whenever you change the type of experiment Refer to the Optimizing the Mass Spectrometer with Your Compound in H ESI MS Mode chapter in your instrument s getting started manual for a procedure to optimize
19. e solvents from Thermo Fisher Scientific go to www FisherLCMS com H ESI Probe User Guide 19 20 5 Replaceable Parts Figure 12 Consumables for the H ESI probe 7 Needle seal P N 97055 20271 H ESI needle P N 97055 20273 Figure 13 PEEK safety sleeve part numbers Fingertight fitting Fingertight fitting Grounding union P N 00101 18195 P N 00101 18081 stainless steel ZVD fitting EIN HQ NT De O Ferrule Ferrule P N 00101 18119 P N 00101 18119 PEEK safety sleeve P N 00301 22806 H ESI Probe User Guide Thermo Scientific Index A APCI probe removing note 7 auxiliary gas flow rate settings 4 5 C calibration frequency note 3 CAUTIONS avoiding an electric shock viii avoiding burns H ESI vaporizer 13 electrical shock PEEK safety sleeve 16 cleaning procedures H ESI probe flushing 13 sample transfer line flushing 13 sample tube flushing 13 compliance WEEE iii daily maintenance H ESI probe flushing 13 sample transfer line flushing 13 sample tube flushing 13 E electrospray rules for good electrospray 3 ESI probe removing note 7 F figures H ESI probe 1 6 H ESI probe exploded view 15 H ESI process 2 installing PEEK safety sleeve 17 installing sample tube 18 ion source interface 1 fused silica sample tube installing 16 installing figure 17 18 trimming 14 Thermo Scientific
20. er die Verwerter und weitere Hinweise die n tzlich sind um die Produkte zu identifizieren die unter diese ROHS Anweisung fallen finden sie unter www thermo com WEEEROHS Conformite DEEE Ce produit doit tre conforme a la directive europ enne 2002 96 EC des D chets d Equipements Electriques et Electroniques DEEE Il est marqu par le symbole suivant Thermo Fisher Scientific s est associ avec une ou plusieurs compagnies de recyclage dans chaque tat membre de l union europ enne et ce produit devrait tre collect ou recycl par celles ci Davantage d informations sur la conformit de Thermo Fisher Scientific ces directives les recycleurs dans votre pays et les informations sur les produits Thermo Fisher Scientific qui peuvent aider la d tection des substances sujettes la directive RoHS sont disponibles sur www thermo com WEEEROHS Contents Thermo Scientific Chapter 1 Chapter 2 Chapter 3 Chapter 4 Chapter 5 it ee vii Safety and Special Notices ner da a Ode oos vii Precautions for Handling the H ESI Probe zu 0 0uuu00s0ua00u 000006 viii Contacting Us ns sus eher ao Goa haw a ix lntrod ction de SS bin dared ee 1 Functional Description 5 Removing and Installing the H ESI Probe 7 Removing the H ESI Probe as san sel 7 salle the H ESI Probe ct hand nasser here 9 Maintenance nina 13 Flushing the Sample Trans
21. fer Line Sample Tube and H ESI Probe 13 Trimming the H ESI Sample Tube us nenn noes 14 Replacing the H EST Needle and Needle Seal 00 2 14 Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve 16 Replaceable Parts 19 Indek rennen 21 H ESI Probe User Guide v Preface This H ESI Probe User Guide provides you with information on using the heated electrospray ionization H ESI probe It also provides procedures for installing and maintaining the H ESI probe CAUTION Read and understand the various precautionary notes signs and symbols contained inside this manual pertaining to the safe use and operation of this product before using the device 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 H ESI Probe User Guide vii Preface Precautions for Handling the H
22. ing Push the sample tube forward so that it extends beyond the end of the H ESI needle Use a fused silica cutting tool to cut off a small length of sample tube Ensure that you cut the end of the sample tube squarely Pull the sample tube backwards until the exit end of the sample tube is recessed just inside the H ESI needle by approximately 0 5 mm between 0 and 1 mm Tighten the sample inlet fitting securely to hold the sample tube Note The sample tube might move forward when you tighten the sample inlet fitting Ensure that the sample tube is retracted into the H ESI needle approximately 0 5 mm between 0 and 1 mm If necessary loosen the fitting and reposition the sample tube Reinstall the H ESI probe as described in the topic Installing the H ESI Probe on page 9 Replacing the H ESI Needle and Needle Seal You must replace the H ESI needle if it is damaged You need to replace the needle seal if the sheath gas is leaking at the needle seal needle interface If you replace the needle you should also replace the needle seal H ESI Probe User Guide To replace the needle and or needle seal see Figure 9 Remove the H ESI probe from the Ion Max source by following the procedure in the topic Removing the H ESI Probe on page 7 Unscrew the sample inlet fitting Remove the sample tube and sample inlet fitting from the H ESI probe Use a 7 64 in hex wrench or ball driver to remove the
23. olution out the nozzle The H ESI probe has inlets for the introduction of sample solution sheath gas auxiliary gas and sheath liquid or calibrant into the H ESI probe The sheath gas is the inner coaxial nitrogen gas that sprays nebulizes the sample solution into a fine mist as it exits the sample tube Typical sheath gas flow rates for H ESI are 5 to 20 units for sample flow rates of less than 10 pL min and 50 to 75 units for sample flow rates greater than 400 pL min When you tune the mass spectrometer for best results adjust the sheath gas flow rate until the ion signal is stable The heated auxiliary gas is the outer coaxial nitrogen gas that assists the sheath gas in the desolvation of sample solutions The auxiliary gas heats as it spirals through a vaporizer The vaporizer is thermally insulated from the sample tube to prevent direct heating of the sample solution You can control the temperature of the vaporizer in the Xcalibur data system The temperature range is from room temperature to 600 C For recommended operating temperatures and gas flow settings see Table 1 on page 4 H ESI Probe User Guide 5 2 Functional Description Figure 3 H ESI probe H ESI vaporizer cable socket Guide pin Pee Sheath gas inlet S AN lt Aux gas inlet A lt J H ESI nozzle H ESI needle high voltage connector receptacle H ESI manifold low Probe depth markers H ESI vaporizer vaporizer shield S
24. perly This document is not part of any sales contract between Thermo Fisher Scientific Inc and a purchaser This document shall in no way govern or modify any Terms and Conditions of Sale which Terms and Conditions of Sale shall govern all conflicting information between the two documents Release history Revision A June 2005 Revision B March 2008 Revision C April 2009 For Research Use Only Not for use in diagnostic procedures WEEE Compliance This product is required to comply with the European Union s Waste Electrical amp Electronic Equipment WEEE Directive 2002 96 EC It is marked with the following symbol Thermo Fisher Scientific has contracted with one or more recycling or disposal companies in each European Union EU Member State and these companies should dispose of or recycle this product See www thermo com WEEERoHS for further information on Thermo Fisher Scientific s compliance with these Directives and the recyclers in your country WEEE Konformitat Dieses Produkt muss die EU Waste Electrical amp Electronic Equipment WEEE Richtlinie 2002 96 EC erf llen 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 entsorgt werden kann Mehr Information ber die Einhaltung dieser Anweisungen durch Thermo Fisher Scientific b
25. re 5 Figure 5 lon Max ion source housing without H ESI probe ELLE H ESI vaporizer cable 8 kV cable ESI interlock socket ESI interlock block m ae Auxiliary gas fitting ESI grounding union holder green Probe locking knob Probe port Probe locking ring Sheath gas fitting blue Thermo Scientific H ESI Probe User Guide 9 3 Removing and Installing the H ESI Probe Installing the H ESI Probe 3 Insert the H ESI probe into the port in the ion source housing Make sure to align the guide pin on the probe body see Figure 6 at a minus 45 angle from the ESI interlock block see Figure 5 Figure 6 H ESI probe front view H ESI vaporizer cable socket Guide pin Sheath gas inlet S Aux gas inlet A H ESI manifold Wy f Probe depth markers 4 Push the probe into the port until the guide pin meets with the locking ring on the ion source housing See Figure 5 5 Turn the probe 45 degrees clockwise and align the guide pin with the slot in the ESI interlock block you might have to pull the probe towards you slightly to properly align the pin with the notch Once you have turned the probe far enough to align the pin with the alignment notch at the rear of the port push the probe straight in until the guide pin stops at the bottom of the alignment notch 6 Lock the probe in place by turning the probe locking knob clockwise See Figure 5 7 Ensure that the grounding union stainless steel ZDV fitting i
26. reassemble the H ESI probe see Figure 9 1 Insert the entrance end of the H ESI needle into the needle seal 2 Reassemble the needle sheath H ESI needle and seal and liquid union junction unit as follows a Seat the H ESI needle and needle seal in the needle sheath b Thread the needle sheath into the liquid junction union Slightly wet the needle sheath threads with HPLC grade methanol for lubrication c With a 3 8 in wrench gently tighten the needle sheath until it is a little more than fingertight Do not overtighten the needle sheath 3 Insert the needle sheath H ESI needle and seal and liquid union junction unit into the H ESI manifold 4 Insert the sheath liquid calibrant adaptor into the H ESI manifold and tighten until fingertight 5 Position the end cover on the H ESI manifold 6 Insert the two 6 32 x 1 4 in socket screws into the H ESI probe and tighten them with a 7 64 in hex wrench or ball driver 7 Carefully insert the fused silica sample tube into the H ESI probe 8 Tighten the sample inlet fitting 9 Reinstall the H ESI probe as described in the topic Installing the H ESI Probe on page 9 Installing a New Fused Silica Sample Tube and PEEK Safety Sleeve H ESI Probe User Guide RL La To install a new sample tube and PEEK safety sleeve CAUTION AVOID ELECTRICAL SHOCK When you are operating your instrument in the H ESI mode you could receive an electrical shock unless you install the
27. rizer 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 H ESI probe Thermo Scientific To remove the H ESI probe Place your mass spectrometer in standby Disconnect the sample transfer tubing LC line from the grounding union stainless steel ZDV fitting Disconnect the 8 kV cable from the H ESI needle high voltage receptacle as follows see Figure 4 a Unlock the cable by twisting the locking ring counterclockwise b Unplug the 8 kV cable from the H ESI needle high voltage receptacle Unplug the H ESI vaporizer cable from the cable socket on the H ESI probe See Figure 4 Connect the H ESI vaporizer cable to the ESI interlock socket on the ion source housing See Figure 5 Disconnect the auxiliary gas fitting green from the auxiliary gas inlet A on the probe manifold See Figure 4 Disconnect the sheath gas fitting blue from the sheath gas inlet S on the probe manifold H ESI Probe User Guide 7 3 Removing and Installing the H ESI Probe Removing the H ESI Probe Figure 4 H ESI probe installed on the lon Max housing H ESI vaporizer cable Auxiliary gas line green fitting Sample inlet red fitting Grounding union 8 kV cable H ESI needle Sheath gas line blue fitting PEEK safety sleeve Probe locking knob So R F ge i
28. s seated in the grounding union holder on the H ESI probe See Figure 7 CAUTION To avoid electric shock ensure that the grounding union is seated in the grounding union holder on the H ESI probe If the grounding union is not seated and you touch the fused silica solvent line while the system is in operation you might receive an electric shock 10 H ESI Probe User Guide Thermo Scientific 10 11 12 13 Thermo Scientific 3 Removing and Installing the H ESI Probe Installing the H ESI Probe Figure 7 H ESI probe rear view H ESI needle high voltage connector receptacle Sample inlet Grounding union holder Sheath liquid calibrant inlet Connect the sheath gas fitting blue to the sheath gas inlet S on the probe manifold See Figure 6 Connect the auxiliary gas fitting green to the auxiliary gas inlet A on the probe manifold See Figure 6 Unplug the H ESI vaporizer cable from the ESI interlock socket Connect the H ESI vaporizer cable to the vaporizer cable socket on the H ESI probe See Figure 6 Connect the 8 kV cable to the H ESI needle high voltage receptacle on the H ESI probe See Figure 7 Tighten the locking ring on the 8 kV connector Connect the sample transfer tubing LC line to the grounding union See Figure 8 H ESI Probe User Guide 11 3 Removing and Installing the H ESI Probe Installing the H ESI Probe Figure 8 H ESI probe installed in the lon Max housing
29. s to download at mssupport thermo com To contact Customer Service for ordering information Phone 800 532 4752 Fax 561 688 8731 E mail us customer support analyze thermofisher com Web site www thermo com ms To copy manuals from the Internet Go to mssupport thermo com and click Customer Manuals in the left margin of the window To suggest changes to documentation or to Help e Complete a brief survey about this document by clicking the link below Thank you in advance for your help SURVEY e Send an e mail message to the Technical Publications Editor at techpubs lcms thermofisher com Thermo Scientific H ESI Probe User Guide ix Introduction Thermo Scientific 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 H ESI can be used to analyze any polar compound that makes a preformed ion in solution The term preformed ion can include adduct ions For example polyethylene glycols can be analyzed from a solution containing ammonium acetate because of adduct formation between the NH ions in the solution and oxygen atoms in the polymer This chapter describes the principles of H ESI and the H ESI probe See Figure 1 The H ESI probe is housed in the Ion Max ion source housing Figure 1 lon source interface left and H ESI probe right With H ESI a mass spectrometer can analyze a range of
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