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CESI 8000 Plus High Performance Separation-ESI Module User Manual RUO-IDV-05-3897-B February 2019...
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This document is provided to customers who have purchased SCIEX equipment to use in the operation of such SCIEX equipment. This document is copyright protected and any reproduction of this document or any part of this document is strictly prohibited, except as SCIEX may authorize in writing.
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Élimination des appareils contenant des composants au mercure, 39 Étiquette marque CE, 39 Étiquette marque MRC, 40 Étiquette marque CSA, 40 CHAPTER 4: System Overview, 41 About This Manual, 41 CESI 8000 Plus High Performance Separation-ESI Module User Manual RUO-IDV-05-3897-B | B11949AF...
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CESI 8000 Plus System: Installing OptiMS Cartridge, 53 OptiMS Cartridge Installation, 53 Thermo Scientific Mass Spectrometer: Integration with the CESI 8000 Plus System, 57 Physical Integration with the CESI 8000 Plus System, 57 OptiMS Cartridge Sprayer Tip Installation into a Thermo...
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Dialog, 118 Insert Option, 120 Sequence Dialog, 121 Sequence Table Rows, 123 Fill Down Option, 127 Display Options, 128 Vial Legend, 129 Tray Detail View, 129 Vial Preview, 130 CESI 8000 Plus High Performance Separation-ESI Module User Manual RUO-IDV-05-3897-B | B11949AF...
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Create a Sequence in the Analyst Software, 173 ® Start a Sequence in the Analyst Software, 176 Load Sample and Reagents on the CESI 8000 Plus System, 177 Sample Tray Setup, 177 Create a Sequence in the CESI 8000 Software, 179...
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Analysis, 214 CHAPTER 10: Shutdown Procedures and Decoupling Instruments, 217 Shutdown and Decoupling Overview, 217 Cartridge Removal and the CESI 8000 Plus System Shutdown, 217 Cartridge Removal and Instrument Shutdown, 217 Decoupling the CESI 8000 Plus System, 222 Decoupling from the Thermo Scientific Mass...
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APPENDIX B: Nominal Network Configuration, 235 Overview, 235 Switching from the CESI 8000 Plus System to Different Mass Spectrometer Systems, 236 Switching to the CESI 8000 Plus System from Different Mass Spectrometer Systems, 236 Switching from a SCIEX Mass Spectrometer to Another Mass...
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Troubleshooting and Flow Diagrams, 349 Sample Preparation Considerations for Analysis with CESI- MS, 350 Properties of a Method or Sequence Cannot be Modified or Saved, 350 Blocked Sprayer Tip, 351 CESI 8000 Plus High Performance Separation-ESI Module User Manual RUO-IDV-05-3897-B | B11949AF...
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Contents Possible Cause, 351 Possible Solution, 351 Electrospray Cannot Be Established Between the CESI 8000 Plus System and Mass Spectrometer, 353 Electrospray is Detected at Zero Ion Spray (IS) Voltage while Applying CE Voltage, 354 Unstable Electrospray while Applying CE Voltage and IS...
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Contents CESI 8000 Plus High Performance Separation-ESI Module User Manual RUO-IDV-05-3897-B | B11949AF...
CHAPTER 1 Safety, Notices and Labels Overview A description of the symbols and labels that are used on the SCIEX CESI 8000 Plus High Performance Separation-ESI Module, or that are shown in this manual, can be found in this section.
Important adds benefit to the performance of a piece of equipment or to a process. NOTE Used to call attention to notable information that should be followed during installation, use, or servicing of this equipment. CESI 8000 Plus High Performance Separation-ESI Module User Manual 14 / 376 RUO-IDV-05-3897-B | B11949AF...
• Acknowledge and act upon instrument alarms and error messages. • Keep away from moving parts. • Report any broken parts to your SCIEX representative. • Use the proper tools when troubleshooting. User Manual CESI 8000 Plus High Performance Separation-ESI Module...
• This equipment is used in a manner other than specified. Operate the instrument as instructed in the product manuals. • You introduce software that is not authorized by SCIEX into your computer. Only operate your system’s computer with software authorized by SCIEX.
• Always wear assigned personal protective equipment, including powder-free neoprene or nitrile gloves, safety glasses, and a laboratory coat. • Follow required electrical safe work practices. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 17 / 376...
This symbol calls attention to important information to read, or is accompanied by another symbol indicating a particular safety hazard. The information is located either on the label with the symbol or in the CESI 8000 Plus documentation. Sharp Object Label This symbol indicates that there are sharp objects, and the operator should use care when accessing this area.
A label cautioning users when moving the cart or adjusting the height of the cart can be found on the CESI 8000 Plus mobile cart. The CESI 8000 instrument is situated on the mobile cart. The OptiMS cartridge spray tip on the CESI 8000 instrument has to be in very close proximity to a mass spectrometer when the system is used.
WEEE (waste, electrical, and electronic equipment). To safely dispose of this equipment, contact a local Customer Service office for complimentary equipment pick- up and recycling. CESI 8000 Plus High Performance Separation-ESI Module User Manual 20 / 376...
The CSA symbol on the CESI 8000 instrument indicates that the instrument has been certified by a Nationally Recognized Testing laboratory (NRTL) to applicable Laboratory Equipment Safety Standards for the United States and Canada. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 21 / 376...
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Safety, Notices and Labels CESI 8000 Plus High Performance Separation-ESI Module User Manual 22 / 376 RUO-IDV-05-3897-B | B11949AF...
合 21 CFR 1040 的适用要求。激光束 不可见。 CLASS 1 LASER PRODUCT THIS PRODUCT CONFORMS TO APPLICABLE REQUIREMENTS OF 21 CFR 1040 AT THE DATE OF MANUFACTURE. MANUFACTURED: 726024-C A016350L EPS CESI 8000 Plus High Performance Separation-ESI Module User Manual 26 / 376 RUO-IDV-05-3897-B | B11949AF...
Présentation Dans cette section, il est possible de trouver une description des symboles et des étiquettes utilisés sur le Module de séparations hautes performances SCIEX CESI 8000 Plus, ou qui sont indiqués dans ce manuel. N’essayez pas d’exécuter une procédure avant d'avoir lu attentivement toutes les instructions.
NOTE Sert à attirer l’attention sur des informations importantes dont il convient de tenir compte lors de l’installation, l’utilisation ou l'entretien de cet équipement. CESI 8000 Plus High Performance Separation-ESI Module User Manual 32 / 376 RUO-IDV-05-3897-B | B11949AF...
• Cet équipement est utilisé d’une manière autre que celle spécifiée Utilisez cet instrument conformément aux instructions des manuels du produit. • Vous installez un logiciel non autorisé par SCIEX sur votre ordinateur. N’utilisez l’ordinateur de votre système qu’avec des logiciels agréés par SCIEX.
Sécurité, Consignes et étiquettes CAUTION Si vous avez acheté ce produit ailleurs que chez SCIEX ou un distributeur SCIEX autorisé, et s’il ne fait pas l’objet d’un contrat de maintenance SCIEX, SCIEX ne peut garantir que le produit a bénéficié des toutes dernières révisions techniques obligatoires ou que vous recevrez les bulletins d’information les...
FABRICATION » se situe à proximité de l'étiquette signalétique. Le faisceau laser n'est pas visible. CLASS 1 LASER PRODUCT THIS PRODUCT CONFORMS TO APPLICABLE REQUIREMENTS OF 21 CFR 1040 AT THE DATE OF MANUFACTURE. MANUFACTURED: 726024-C User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 35 / 376...
Symbole « Haute tension, risque de choc électrique » Ce symbole indique la présence de haute tension et d'un risque de choc électrique, et l'opérateur doit faire preuve de précautions lors de l'accès à cette zone. CESI 8000 Plus High Performance Separation-ESI Module User Manual 36 / 376...
Ce symbole attire l'attention sur des informations importantes à lire, ou est accompagné d'un autre symbole indiquant un danger particulier pour la sécurité. Les informations se trouvent soit sur l'étiquette avec le symbole ou dans la documentation du CESI 8000 Plus. Étiquette Objet tranchant Ce symbole indique la présence d'objets tranchants et l'opération doit faire preuve de...
être indiqués par ces étiquettes et symboles et sont précisés dans les Mises en garde ou les Avertissements dans les procédures décrivant la tâche. CESI 8000 Plus High Performance Separation-ESI Module User Manual 38 / 376...
Marque CE - La marque « CE » indique qu’un produit a été évalué avant d’être placé sur le marché et qu’il a été jugé comme répondant aux exigences de l’Union européenne en matière de sécurité, de santé et/ou de protection de l’environnement. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 39 / 376...
Nationally Recognized Testing laboratory (NRTL - laboratoire national d’essai reconnu) aux Normes sur la sécurité de l’équipement de laboratoire pour les États-Unis et le Canada. CESI 8000 Plus High Performance Separation-ESI Module User Manual 40 / 376 RUO-IDV-05-3897-B | B11949AF...
• A photo diode array (PDA) detector that is in the 190 nm to 600 nm range. • A laser-induced fluorescence (LIF) detector that permits high-sensitivity analysis of labeled molecular species. For instructions on using the UV, PDA, or LIF detector, go to the SCIEX web site at www.sciex.com and then click Products >...
CE in an ultra low-flow format that enhances the sensitivity of mass spectrometry while reducing ion suppression. Within the CESI 8000 Plus system, sample components are separated by an electrical field that is applied between the inlet vial and the OptiMS sprayer (Figure 4.1).
System Overview This manual will describe the major components of the CESI 8000 Plus system with OptiMS technology, and common procedures such as installation, integration with mass specs, as well as the more commonly used software procedures. Appendices describe care and maintenance of CESI 8000 Plus components, and corrective actions for specific problems that may occur.
High voltage (HV) from the mass spectrometer connects to the current monitor circuit on the CESI 8000 Plus system through the input HV cable. An output cable delivers the voltage from the outlet side of the current monitor circuit to the OptiMS capillary, required for the electrospray.
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Wear protective eyewear, lab coat, and gloves when opening the sample cover. The CESI-MS vials are pressurized during rinse and separation-with-pressure events. To reduce the risk of breakage and expelled particles, use only SCIEX vials designed for the CESI 8000 Plus system (PN B11648). Inspect every vial for damage before use.
Syringe Pump, Power Supply, LEDs and Interlock Syringe Pump The CESI 8000 Plus system can generate pressures with an internal pump mechanism. This pump can supply 0.1 psi to 25 psi to perform pressure injections or low-pressure mobilizations. The pump can apply a maximum of 100 psi to move the fluids through the capillary. The pressure can...
System Overview Cartridge and Sample Cover Interlocks The hinged doors of the CESI 8000 Plus system have interlock sensors that prevent unsafe access to the inside of the instrument. The outer door is called the sample cover; the second inner door is called the cartridge cover.
4.8). These capillaries are shrouded at the inlet ends by protective covers which automatically retract when the OptiMS cartridge is installed in the CESI 8000 Plus system. The spray end (5) is made of PEEK, which is a high performance plastic material that has been tested and proven to be inert under CESI-MS conditions.
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OptiMS cartridge is installed in the CESI 8000 Plus system. The locking mechanism will not retract if the OptiMS cartridge is not installed on the CESI 8000 Plus system.
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System Overview • OptiMS adapter for SCIEX mass spectrometer (Figure 4.10) Figure 4.10 Adapter for SCIEX Nanospray® III Ion Source 1. HV Input Cable 2. HV Output Cable • OptiMS adapter for Thermo Scientific mass spectrometer (Figure 4.11) Figure 4.11 Adapter for Thermo Scientific NanoSpray II ion source 1.
Integrating the CESI 8000 Plus System with a Mass Spectrometer CESI-MS Integration Procedures This chapter discusses the integration of the CESI 8000 Plus system with the following mass spectrometers: • Thermo Scientific LTQ mass spectrometer series • Thermo Scientific Q Exactive mass spectrometer ®...
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WARNING To prevent a shock hazard, do not touch the mass spectrometer adapter when connected to the CESI 8000 Plus system and the separation voltage is on. CESI 8000 Plus High Performance Separation-ESI Module User Manual 52 / 376...
Integrating the CESI 8000 Plus System with a Mass Spectrometer CESI 8000 Plus System: Installing OptiMS Cartridge The OptiMS interface plate must be installed onto a CESI 8000 Plus system before an OptiMS cartridge can be installed. Refer to Removing and Re-installing OptiMS Interface Plate...
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Integrating the CESI 8000 Plus System with a Mass Spectrometer To install the cartridge body, sit the cartridge’s ends on the aligning blocks of the OptiMS interface plate at a 45 degree angle, and lightly push it in (Figure 5.3).
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.4 Pivot Cartridge Upwards and Align onto Interface Plate 1. Align “T” Slider into the Groove Figure 5.5 Slide Cartridge All the Way Down 1. Capillaries (appear beneath interface block and within the ejectors)
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Route coolant and capillary tubing through the notched arm (Figure 5.6). Figure 5.6 Routing Coolant and Capillary Tubing Bring the lock down bar to the front of the cartridge and secure cartridge in place by...
Ensure that both the CESI 8000 Plus system and controller are connected to electrical power. Connect the GPIB-USB cable to the connector labeled “Controller” on the CESI 8000 Plus system. Plug the other end of the GPIB-USB cable to a USB port on the CESI 8000 Plus controller (Figure 5.8).
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Connect the relay cable to the relay input (labeled I/O) on the CESI 8000 Plus system. Connect the other end of the relay cable to the Thermo Scientific mass spectrometer relay...
To prevent a shock hazard, ensure that the MS ESI voltage is set to zero and idle. The CESI 8000 Plus system separation voltage should be off. Prior to installing the NanoSpray II ion source, ensure the 1-inch adapter ring is not installed (Figure 5.11).
Integrating the CESI 8000 Plus System with a Mass Spectrometer Additionally, ensure the ion sweep cone has been removed from the mass spectrometer (Figure 5.12). CAUTION The ion sweep cone may be hot. Care must be taken when manipulating it.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer WARNING It is critical to perform Step 2 prior to installation of the adapter. During the installation of the OptiMS spray assembly, the glass tip of the OptiMS sprayer will be exposed. If the ion source stage is too close to the mass spectrometer inlet, then damage to the tip may result.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.15 Adapter for Thermo Scientific Mass Spectrometer in Place 1. Thumbscrew 2. Adapter (in place) Remove protective sleeve from OptiMS sprayer’s tip (Figure 5.16). IMPORTANT Save the protective sleeve. It will be needed if the OptiMS cartridge has to be removed and/ or stored for later use.
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WARNING To prevent a shock hazard, ensure that the MS ESI voltage is set to zero and idle. The CESI 8000 Plus system separation voltage should be off. Figure 5.18 HV Cable to Mass Spectrometer Input Cable (Cable-to-Cable) Connection User Manual...
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Integrating the CESI 8000 Plus System with a Mass Spectrometer To expose the HV connectors, slide open the door located on the lower-left corner of the right-side panel of the CESI 8000 Plus system (Figure 5.19). Connect the black ground banana plug from the cable to the matching black receptacle on the panel.
Integrating the CESI 8000 Plus System with a Mass Spectrometer WARNING To prevent a shock hazard, install the HV Input and Output cables and then check the connections at both ends before turning on the CESI 8000 Plus system. Figure 5.20 High Voltage Panel Connections 1.
Aligning the CESI 8000 Plus System with a Thermo Scientific Mass Spectrometer The level of the buffer vials in the CESI 8000 Plus system must be at the same level as the OptiMS cartridge sprayer tip. This will prevent hydrodynamic (siphoning) flow within the capillaries, which may affect CE-MS separation performance.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer If necessary, use the knob shown in Figure 5.23 to retract the ion source stage all the way back so that the sprayer tip does not collide with the mass spectrometer when moving the stage.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer WARNING It is critical to perform Step 1 prior to moving the CESI 8000 Plus cart towards the stage. This will avoid any potential damage to the tip of the OptiMS sprayer.
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Always press the height adjustments buttons one at a time, never simultaneously. IMPORTANT If the CESI 8000 Plus cart will not power to its lowest height, or if it becomes stuck, then the cart must be re-programmed. Refer to the...
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.26 Cart Height Adjustment Buttons 1. Cart Up Button 2. Cart Down Button Use the mass spectrometer alignment knobs to make fine adjustments to align the OptiMS sprayer tip to the center of the source inlet on the mass spectrometer (Figure 5.27).
Integrating the CESI 8000 Plus System with a Mass Spectrometer When the CESI 8000 Plus system is visually aligned to the mass spectrometer, lock the wheels on the CESI 8000 Plus cart by stepping on the locking lever at each caster (Figure 5.28).
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Connect the GPIB-USB cable to the connector labeled “Controller” on the CESI 8000 Plus system. Use spacer if needed. Plug the other end of the GPIB-USB cable to a USB port on the...
To prevent a shock hazard, ensure that the MS ESI voltage is set to zero and idle. The CESI 8000 Plus separation voltage should be off. To install the mass spectrometer spray end of the OptiMS cartridge into the SCIEX Nanospray® III source, perform the following steps: Install the SCIEX Nanospray®...
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.32 Pulling Back the SCIEX Assembly WARNING It is critical to perform Step 2 prior to installation of the adapter. During the installation of the OptiMS spray assembly, the glass tip of the OptiMS sprayer will be exposed.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.33 Orientation of the Adapter Prior to Installation onto the Ion Source 1. High Voltage (HV) Output Cable ® 3. SCIEX Nanospray III Adapter 2. Hook 4. SCIEX Nanospray ®...
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Remove protective sleeve from the OptiMS sprayer tip (Figure 5.35). IMPORTANT Save the protective sleeve. It will be needed if the OptiMS cartridge has to be removed and/ or stored for later use.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer The OptiMS has a locking mechanism which keeps the OptiMS Spray Assembly securely attached to the adapter. Twist the OptiMS sprayer assembly counter clockwise (Figure 5.37) to lock it into place within the adapter.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.38 CESI 8000 Plus System HV Panel WARNING To prevent a shock hazard, ensure that the MS ESI voltage is set to zero and idle. The CESI 8000 Plus separation voltage should be off.
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Connect the red banana ground plug into the matching red receptacle on the panel. Connect the HV cable from the mass spectrometer ion source into the input connection on the CESI 8000 Plus HV panel, and twist 90 degrees clockwise to lock it in place (Figure 5.39 Figure 5.40).
Integrating the CESI 8000 Plus System with a Mass Spectrometer Establishing Communication Between the CESI 8000 Plus Controller and the SCIEX Mass Spectrometer The CESI 8000 Plus controller (PC) communicates with the SCIEX mass spectrometer using an Ethernet cable (Figure 5.30).
5600 Mass Spectrometer The level of the buffer vials in the CESI 8000 Plus system must be at the same level as the OptiMS cartridge sprayer tip. This will prevent hydrodynamic (siphoning) flow within the capillaries, which may affect CE-MS separation performance. This leveling is done by aligning the height indicator line on the CESI 8000 Plus system with the OptiMS sprayer tip placed at the mass spectrometer adapter.
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Integrating the CESI 8000 Plus System with a Mass Spectrometer Figure 5.43 Move Stage All the Way Back 1. Coarse Z-axis Adjustment Knob 2. Stage IMPORTANT Make sure that the Source Drain assembly on the mass spectrometer is covered. The...
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If the ion source stage is too close to the mass spectrometer inlet, then damage to the tip may result. Move the CESI 8000 Plus cart close, approximately 4 inches (10 cm.) to the ion source stage so that the OptiMS coolant tubes can be fully extended (Figure 5.45).
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Always remove consumable before adjusting the CESI 8000 Plus cart height. Use the up and down height-adjustment buttons on the front of the cart to adjust the height of the cart until the height indicator on the CESI 8000 Plus system is aligned to the OptiMS sprayer tip (Figure 5.47).
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Always press the height adjustments buttons one at a time, never simultaneously. IMPORTANT If the table top of the CESI 8000 Plus cart will not power to its lowest height, or if it becomes stuck, then the cart must be re-programmed. Refer to the...
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When the CESI 8000 Plus system is in visual alignment to the mass spectrometer, lock the wheels on the CESI 8000 Plus cart. This is done by stepping on the locking lever at each caster to lock each wheel in place (Figure 5.49).
The 32 Karat software is a full-featured chromatography and capillary electrophoresis software package that provides control of the CESI 8000 Plus system. For the CESI 8000 Plus user, there is no electrophoretic data collected by the 32 Karat software. Only voltage, electrical current, pressure, and power data traces can be collected.
If the instrument is installed on a network, the user name and password will be supplied by the network administrator. 32 Karat software functionality is different when run on a network CESI 8000 Plus High Performance Separation-ESI Module User Manual...
Service Support selection. Instrument Start Up The software can only control the CESI 8000 Plus system after it has completed its startup initialization process. Before starting the CESI 8000 Plus system: • Make sure the cartridge is installed.
An instrument is opened online by double-clicking the instrument icon. Open Offline To open an instrument offline, right-click the instrument icon and select CESI 8000 Plus High Performance Separation-ESI Module User Manual 90 / 376 RUO-IDV-05-3897-B | B11949AF...
Using the Direct Control Window to Load the Cartridge and Samples Direct Control Control can be selected from the menu (Figure 6.5). Figure 6.5 Control Menu for Direct Control User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 91 / 376...
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(Figure 6.8). Figure 6.8 Direct Control Window Load Select to move the trays to the front of the sample compartment. CESI 8000 Plus High Performance Separation-ESI Module User Manual 92 / 376 RUO-IDV-05-3897-B | B11949AF...
® ® communicate with the Analyst TF software used by the controller of the SCIEX TripleTOF 5600 mass spectrometer. If AAO is being used, the Direct Control window will display an icon that indicates the mass spectrometer status of the AAO interface (Figure 6.9).
® For AAO configurations (such as for the SCIEX TripleTOF 5600 mass spectrometer), the relay event will echo back to the software to send the start acquisition command through the AAO interface.
CESI 8000 Software Online Help. Instrument Setup — Initial Conditions Initial Conditions tab is used to set instrument parameters at the start of a method, before the separation begins. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 95 / 376...
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Wait for external trigger a run. These are selected by selecting the appropriate box. If is selected, the CESI 8000 Plus system becomes a slave device and does not start until an external signal is Wait for temperature received. The two options assure that the system has reached the correct operating temperature before beginning a run.
Some events do not have a time option, others can be timed or untimed. Collection of voltage and current data begins with the first User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 97 / 376...
Figure 6.12 Time Program Event List The following dialogs and field definitions are the ones that should be more commonly used by the CESI 8000 Plus user. For information on more advanced event types, refer to the CESI 8000 Software Online Help.
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Figure 6.14 Tray Selection Dialog User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 99 / 376...
The inject event is used to deliver a precisely measured amount of sample into the capillary (Figure 6.16). This step is always untimed, and usually precedes the first separation step. CESI 8000 Plus High Performance Separation-ESI Module User Manual 100 / 376...
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The Allow Override box must be checked in the Sequence Table area (in Figure 6.16) to give the sequence parameters priority over the method parameters. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 101 / 376...
For example, type: TestMethod_111518 Click Save C:\32Karat\Projects\Default\Methods The default path at installation is: Your system administrator may have assigned you to a different default path. CESI 8000 Plus High Performance Separation-ESI Module User Manual 102 / 376 RUO-IDV-05-3897-B | B11949AF...
Sequence mode is used to append multiple methods together as a sequence. The CESI 8000 Plus system must be online to run samples and acquire data. If you are not online, “offline” appears in the title bar of the instrument window. Close any offline windows before proceeding.
, select For Acquisition . Amount Values are not used in this example. Next Click . The Sequence Wizard — Unknowns page displays (Figure 6.21). CESI 8000 Plus High Performance Separation-ESI Module User Manual 104 / 376 RUO-IDV-05-3897-B | B11949AF...
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Data Path Click the green folder icon, and select , then navigate to the folder. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 105 / 376...
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Number of Unknown Runs in Sequence fields. Next Click . The Sequence Wizard — Vials page displays (Figure 6.24). Figure 6.24 Sequence Wizard — Vials Page CESI 8000 Plus High Performance Separation-ESI Module User Manual 106 / 376 RUO-IDV-05-3897-B | B11949AF...
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Figure 6.25 Sequence Wizard — Reports Page This page (Figure 6.25) allows you to have reports generated automatically at the end of the sequence run. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 107 / 376...
1, rep 2, etc., appended to the file name. Sample Inject Inlet — Indicates the vial the sample is being injected from if sample is located in the inlet tray. CESI 8000 Plus High Performance Separation-ESI Module User Manual 108 / 376...
32 Karat Software Overview Sample Inject Outlet — Indicates the vial the sample is being injected from if sample is located in the outlet tray. In the CESI 8000 Plus system the sample is always located in the inlet sample tray.
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32 Karat Software Overview CESI 8000 Plus High Performance Separation-ESI Module User Manual 110 / 376 RUO-IDV-05-3897-B | B11949AF...
32 Karat software, the CESI 8000 software has been developed with the routine user in mind. The CESI 8000 software steps the instrument operator through the tasks of running methods and sequences on the CESI 8000 Plus system. Routine users often receive Standard Operating Procedures detailing separation methods, sequences, and report templates developed in the 32 Karat software by method developers.
CESI 8000 Software Launching the CESI 8000 Software Launch the CESI 8000 software by double-clicking the CESI 8000 Plus icon on the Windows desktop. The Ready window appears as shown in Figure 7.1. The upper right buttons activate the major functions. Hover the mouse over the buttons to obtain a balloon tip regarding function.
Figure 7.2 Login Dialog NOTE If 32 Karat software security is configured to use the Domain controller, then the system will also prompt for the Domain name. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 113 / 376...
When the CESI 8000 software is locked, other users will still be able to access other programs (such as the 32 Karat software) on the system. About Dialog To view the software versions for the CESI 8000 Plus, 32 Karat, and the instrument firmware Copy Version versions, select .
• Application Selection • Samples and Vials • Run Acquisition Application Selection The first step of Run Application is to select an application (Figure 7.3). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 115 / 376...
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Instead of using the 32 Karat button in this instance, use the “Show 32 Karat” button at the center bottom of the dialog. CESI 8000 Plus High Performance Separation-ESI Module User Manual 116 / 376...
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(bring the trays into the Home or Load position). Click Next or select to prepare the samples and vials for the selected application, see Samples and Vials. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 117 / 376...
The tray detail for the method is shown. To set up a sequence, see Sequence Dialog. Figure 7.7 Method Setup Window CESI 8000 Plus High Performance Separation-ESI Module User Manual 118 / 376 RUO-IDV-05-3897-B | B11949AF...
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A dialog will appear to confirm that the samples are loaded. Verify that they are loaded, and Yes - run now select to start the run, see Acquisition. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 119 / 376...
Figure 7.9 Insert Sample ID Dialog Figure 7.10 Insert Data File Dialog Options for the Sample ID (Figure 7.9) or Data File (Figure 7.10) dialogs include: CESI 8000 Plus High Performance Separation-ESI Module User Manual 120 / 376 RUO-IDV-05-3897-B | B11949AF...
Describe Sequence dialog by selecting . If the sequence is modified and saved in the Describe Sequence dialog, select Reload Sequence to reload the described sequence. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 121 / 376...
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Describe Sequence. NOTE When a particular run is selected, the vials used in the run are highlighted in the tray view. CESI 8000 Plus High Performance Separation-ESI Module User Manual 122 / 376 RUO-IDV-05-3897-B | B11949AF...
— Never hidden and always run, regardless of the number of samples (Figure 7.13). NOTE When the number of samples is set to 0, all rows are hidden except for rows set to Always. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 123 / 376...
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CESI 8000 Software Figure 7.12 Sequence with All 9 Samples Figure 7.13 Sequence with No Samples (All Rows Hidden) CESI 8000 Plus High Performance Separation-ESI Module User Manual 124 / 376 RUO-IDV-05-3897-B | B11949AF...
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7.14) to four (Figure 7.15). Hidden rows that are not run can be shown by right-clicking the sequence and selecting Show Hidden Rows (Figure 7.16 Figure 7.17). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 125 / 376...
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Figure 7.17 Sequence with Four Samples and Hidden Rows For more information on setting row types, see Step 6 of Describe Sequence. For more information on the number of samples see Sequence Dialog. CESI 8000 Plus High Performance Separation-ESI Module User Manual 126 / 376 RUO-IDV-05-3897-B | B11949AF...
— (Data File only) The current Sample ID for the Data file. Sample ID • — The current user logged into the application. User Name User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 127 / 376...
Vial Legend. NOTE The vial legend can be shown or hidden from the Display Options menu or by clicking the button highlighted in Figure 7.21. CESI 8000 Plus High Performance Separation-ESI Module User Manual 128 / 376 RUO-IDV-05-3897-B | B11949AF...
Vial Contents). From this dialog, the user can view, export, or print the current trays. The vials can be viewed as circles or rectangles. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 129 / 376...
Cycle number is the number of times that the specific method has been run. This window is used to show tray collisions (marked in red). Figure 7.23 Sequence Vial Preview CESI 8000 Plus High Performance Separation-ESI Module User Manual 130 / 376...
The left-hand panel has three tables to show different levels of detail about a method or sequence. The Run Queue table is used to verify that the run initiated in CESI 8000 Plus system is in the 32 Karat software Run Queue. If a sequence is running, the Sequence Run table will show which row of the sequence is currently running.
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Start Now to cancel the waiting and start the run. Click Stop to abort the scheduled run, and the system state will switch to Run Aborted CESI 8000 Plus High Performance Separation-ESI Module User Manual 132 / 376 RUO-IDV-05-3897-B | B11949AF...
Mass spectrometer interface is Offline or Disconnected. Mass spectrometer is on Standby, Stopping, Paused, Unknown or AAO driver connection failed. Mass spectrometer is Ready. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 133 / 376...
(select to help distinguish between the traces). • — Changes the line thickness of the trace. Thin Lines CESI 8000 Plus High Performance Separation-ESI Module User Manual 134 / 376 RUO-IDV-05-3897-B | B11949AF...
Passwords to ensure system security. The CESI 8000 software is optimized for use with a user- login enabled system. Sequence Transfer Define an Instrument for the Routine User Start the 32 Karat software and login as an Administrator. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 135 / 376...
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Define the Project for the Instrument Tools > System Administration Wizard Select In the System Administration wizard, select Project and then select Next CESI 8000 Plus High Performance Separation-ESI Module User Manual 136 / 376 RUO-IDV-05-3897-B | B11949AF...
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Enter the User Name and Password, select Select Select Tools > System Administration Wizard User Next Select and select Select the User Name and select Next User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 137 / 376...
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Select the Domain and then enter the User Name. Select Check Names . Add the name to the Selected Users by selecting Next Select CESI 8000 Plus High Performance Separation-ESI Module User Manual 138 / 376 RUO-IDV-05-3897-B | B11949AF...
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32 Karat project for routine use on existing systems. As a result, methods may already be in the project folder. Transfer the Sequences Copy the sequences into the project’s Sequence folder. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 139 / 376...
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Ensure that the methods are selected from the corresponding method folder for this project (Figure in the Sequence Run Table 7.29). Verify that any file paths in the Method column are correct. CESI 8000 Plus High Performance Separation-ESI Module User Manual 140 / 376 RUO-IDV-05-3897-B | B11949AF...
The sequence will need to be re-described (using Describe Sequence) in order to vary the sample quantity. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 141 / 376...
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This example is for an instrument named QC001 and a 32 Karat project named QC001. Figure 7.31 Describe Sequence Start Dialog Application In the field, select the application. In the Sequence field, select the sequence to describe. CESI 8000 Plus High Performance Separation-ESI Module User Manual 142 / 376 RUO-IDV-05-3897-B | B11949AF...
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Show hidden rows . For more information on how rows are hidden based upon the number of samples see Sequence Table Rows. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 143 / 376...
• Program Event — Changes the view of the trays to show the action performed on the vial (for example, separation, rinse, sample inject, etc.). CESI 8000 Plus High Performance Separation-ESI Module User Manual 144 / 376 RUO-IDV-05-3897-B | B11949AF...
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Show Vial Preview — Opens a table to preview the methods and vials used in the method/ sequence and shows tray collisions, see Vial Preview. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 145 / 376...
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CESI 8000 Software CESI 8000 Plus High Performance Separation-ESI Module User Manual 146 / 376 RUO-IDV-05-3897-B | B11949AF...
Stock Reagents and Other Consumables Required • Preparation of Reagent Solutions and Test Sample • Procedure for Installation of Adapter and OptiMS Cartridge • Methods for the CESI 8000 Plus System • Capillary Conditioning • Establishing a Stable Spray and Determining Optimum ESI Voltage •...
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System The workflow is as follows: Before you start, ensure you have all the required reagents properly prepared. Description and preparation protocols can be found in the next section.
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Table 8.2 Materials (Continued) Consumables Vendor Part Number 50 mL volumetric flask 20 mL glass bottle 100 mL glass bottle pH meter Table top mini centrifuge...
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Preparation of 10% (v/v) HAc—Background Electrolyte (BGE) NOTE This solution must be prepared fresh every day. IMPORTANT Sonicate in a water bath for 30 minutes prior to use.
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Adjust the pH of the solution to 4.0 by adding aliquots of freshly prepared 20% HAc. Transfer the solution into a clean 100 mL volumetric flask.
CESI 8000 Plus methods described in this chapter. NOTE Use only CESI-MS vials and CESI-MS caps in the CESI 8000 Plus system buffer and sample trays. Table 8.4 provides the vial content, quantity, and volume for capillary conditioning beta- galactosidase auto-calibration, manual calibration, and system test methods.
The sample tray setup is described in this section. It is important to follow the specific location for each sample as they match the CESI 8000 Plus methods described in this chapter. The beta-galactosidase sample used for the auto-calibration and the beta-galactosidase sample...
For installation instructions for the adapter and the OptiMS cartridge, refer to CHAPTER 5, Integrating the CESI 8000 Plus System with a Mass Spectrometer within this manual. Prior to inserting the OptiMS sprayer into the adapter, ensure the positioning stage is aligned as...
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.4 Pull back Positioning Stage from the SCIEX Mass Spectrometer For additional information, refer to the OptiMS Cartridge Sprayer Tip Installation into the SCIEX Nanospray® III Source...
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The CESI-MS Separation (beta-galactosidase) is a method used to inject and separate a beta- galactosidase control sample by CESI-MS. This method is used to ensure both the CESI 8000 Plus ®...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System The time program for all 5 methods differ in a variety of steps; Figure 8.6 through Figure 8.8 show how each time program is set. Figure 8.6 shows the time program settings for the CESI-MS Capillary Conditioning method.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.7 shows the time program settings for the CESI-MS Auto-calibration method to be used in a batch run requiring auto-calibration. Figure 8.7 CESI-MS Auto-calibration_ABSciex Method Settings...
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System The CESI-MS Cleaning method used at the end of every sequence is shown in Figure 8.10. Refer APPENDIX F for detailed information. Figure 8.10 CESI-MS Cleaning Method Settings...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System IMPORTANT If creating a method from scratch, the following applies: 1. Synchronization Mode is set to No Sync. 2. Ensure that the Beckman CE Driver is present on the left pane under Acquisition Method.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Go to Advanced MS tab to enter other parameters (Figure 8.14). Figure 8.14 Parameter Settings for Advanced Mass Spectrometer Tab Click Product Ion (+) to enter the mass spectrometer parameters for Product Ion scan (Figure 8.15).
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Click Edit Parameters , and a dialog for Source/Gas Compound will appear. Enter parameters in both tabs (Figure 8.16). Figure 8.16 Parameter Settings for Source/Gas and Compound tabs of Product Ion Scan Save the method as CESI_MS_AB_Sciex_Betagal_45min.dam...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Capillary conditioning is a procedure that must be performed when using an OptiMS Silica Surface Cartridge for the first time, and when using a cartridge that has been stored for a period longer than 3 days.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Click the icon to open 32 Karat software. Direct Control In 32 Karat software, click the icon on the toolbar (Figure 8.19). Figure 8.19 Direct Control Icon on the Toolbar...
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.21 Single Run Acquisition Dialog The CESI 8000 software will start the series of rinses and at the completion of the initial capillary conditioning method the OptiMS capillary cartridge is ready for the next step, establishing a stable spray.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System ® on the SCIEX TripleTOF 5600 mass spectrometer, refer to the Aligning the CESI 8000 Plus System ® with the SCIEX TripleTOF 5600 Mass Spectrometer section within CHAPTER 5 of this manual.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.24 Droplet at the Stainless Steel Needle End To fill the separation capillary with background electrolyte (10% Acetic acid) follow these two easy steps: Go to the Direct Control window, click...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.26 Droplet at the Tip of the Sprayer WARNING As droplets form at the end of the sprayer tip, ensure that are not aspirated into the mass spectrometer inlet. Do not position the sprayer tip closer than 2 mm from the curtain gas plate.
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However, CE-MS runs use a curtain gas value of 5. It has been set (factory setting) to 5 when the ® TripleTOF 5600 mass spectrometer is interfaced with CESI 8000 Plus mass spectrometer. User Manual CESI 8000 Plus High Performance Separation-ESI Module...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System From the Scan type list, select TOF MS , set the Accumulation time to 500 ms , and set the TOF 1500. 5 min Masses (Da) to Min.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System ® In the Analyst TF software, go to the Advance MS tab and check the box. Go back to the Accumulation time 1 (sec) Duration 5.00 min...
Running CESI-MS ® This section provides a tutorial on the use of the CESI 8000 Plus system with the SCIEX TripleTOF 5600 mass spectrometer. The tutorial uses a tryptic digestion of beta-galactoside as the sample. The beta-galactosidase peptide digest is a standard mixture commonly used to evaluate mass spectrometer performance and it will be used here to evaluate the performance of the capillary separation as well.
This section provides step by step instructions on how to build a sequence of 10 runs of beta- ® galactosidase performance test on the TripleTOF 5600 mass spectrometer with auto-calibration every 5 runs in tandem with the CESI 8000 Plus system. In the navigation bar, under Acquire , double-click...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.33 Add Sample Dialog Click to close the Add Sample dialog and the Batch Editor opens again with all the Vial Position information entered. The...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.35 Batch Editor Window — Calibrate Tab View Click to ensure that the correct reference table is selected. There are 8 reference ions used for mass spectrometer calibration and the peptide at 729.3 m/z is selected for MS/MS calibration.
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System ® Start a Sequence in the Analyst TF Software Click Submit to submit the batch. This will submit the 10 beta-galactosidase separation runs with auto-calibration (Figure 8.37).
The sample tray setup is described in this section. It is important to follow the specific location for each sample as they match the CESI 8000 Plus methods described in this chapter. The beta- galactosidase sample used for the auto-calibration and the beta-galactosidase sample used for...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System IMPORTANT Samples are likely to evaporate over time. It is recommended to use a minimum of 50 μL of sample for a sequence with a duration of approximately 24 hours. If the duration of the sequence is over 50 hours, a minimum volume of 80 μL is strongly recommended.
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Create a Sequence in the CESI 8000 Software In the CESI 8000 software, go to Menu > Sequence > Sequence Wizard Sequence Wizard — Method Method...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Fill in the fields for the Sample ID Data path , and Data file . By clicking on the blue arrow next to the Data file field, a series of options come up to increment the file name. In this example,...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.43 Sequence Table IMPORTANT The autocal mix is always on sample inlet tray position A1, and in this example the sample is on position A2. Ensure that the sample position is designated properly in the sample table before submitting the sequence to run.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.45 Sample Sequence Example NOTE Auto-calibration data file names must be different. In the toolbar, go to Sequence > Properties . In the Sequence Properties dialog (Figure 8.46)
® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Start a Sequence in the CESI 8000 Software To run the sequence, click the Run Sequence icon and the Run Sequence dialog will appear Start (Figure 8.47).
20 kV. Automatic Rinse Feature The CESI 8000 Plus system makes use of an “Automatic Rinse” feature. In the event of a hardware failure during the rinse steps as well as the injection step, either during a method run or sequence run, the instrument will (without user intervention) rinse the separation capillary for 3 minutes at 50 psi from the first separation event vial position.
(in 100 mM LE separated using 10% HAc as background electrolyte), is shown in Figure 8.49. * Ref. LC/MS Peptide/Protein Mass Standards Kit for Tuning of the AB SCIEX ESI Instrument Protocol. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.49 Total Ion Chromatogram (TIC) Create the extracted ion chromatogram (XIC) for peaks 729.3652 m/z and 714.8469, both with a width of 0.05. IMPORTANT The peak range 729.3652 m/z and 714.8469 m/z may need to be adjusted based on the mass spectrometer mass calibration.
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System Figure 8.50 Extracted Ion Chromatogram (XIC) and Mass Spectra for a Beta-Galactosidase Run User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 187 / 376...
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® Using a SCIEX TripleTOF 5600 Mass Spectrometer with the CESI 8000 Plus System CESI 8000 Plus High Performance Separation-ESI Module User Manual 188 / 376 RUO-IDV-05-3897-B | B11949AF...
Procedure for Installation of Adapter and OptiMS Cartridge • Capillary Conditioning • Establishing a Stable Spray and Determining Optimum ESI Voltage • Tuning • Running CESI-MS The workflow is as follows: User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 189 / 376...
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Before you start, ensure you have all the required reagents properly prepared. Description and preparation protocols can be found in the next section. Stock Reagents and Other Consumables Required Table 9.1 Stock Reagents...
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Preparation of Reagent Solutions and Test Sample Table 9.3 Required Reagents Reagents Concentration 10% Acetic Acid (v/v) 10% HAc (v/v) Background electrolyte and conductive liquid 20% Acetic Acid (v/v)
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Preparation of 20% (v/v) Acetic Acid (HAc) — For LE Buffer pH Adjustment NOTE Prepare fresh and discard after use. Do not stock this solution. Add 80.0 mL of DDI water into a clean 100 mL glass bottle.
A1, as shown Figure 9.1. Samples to be analyzed can be placed in other positions; however, these positions must be specified in the CESI 8000 Plus methods or manually specified in the test sequence. User Manual CESI 8000 Plus High Performance Separation-ESI Module...
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System IMPORTANT Samples are likely to evaporate over time. It is recommended to use a minimum of 50 μL of sample for a sequence with a duration of approximately 24 hours. If the duration of the sequence is over 50 hours, a minimum volume of 80 μL is strongly recommended.
CESI 8000 Plus methods described in this chapter. NOTE Use only CESI-MS vials and CESI-MS caps in the CESI 8000 Plus system buffer and sample trays. Table 9.4 provides the vial content, quantity and volume needed for capillary conditioning, and system performance test methods.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Table 9.4 Solution/Reagent Requirements Vial Content Quantity Volume DDI water 2 vials 1.5 mL 0.1 N NaOH 1 vial 1.5 mL 0.1 N HCl 1 vial 1.5 mL...
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Prepare inlet and outlet buffer trays as shown in Figure 9.1. Then set the trays aside. Open the CESI 8000 software. Click the icon to open the 32 Karat software.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Figure 9.6 shows the dialog for the single run. Under Run information field, confirm the method path and name are correct for capillary conditioning. It is not necessary to enter the sample ID nor data file name as no data file will be generated.
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Prior to establishing a stable spray, it is important the sprayer tip position is optimized relative to the mass spectrometer inlet. For detailed information on how to adjust and align the sprayer...
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System As the conductive liquid capillary fills with background electrolyte, a droplet is observed at the end of the stainless steel needle. After the rinse step completes, proceed to the next step to fill the separation capillary.
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Figure 9.10 Voltage Settings The CESI 8000 software electrical current should be around 2 mA to 3 mA. While the CESI 8000 Plus separation voltage is on, apply ESI voltage in increments of 100 V starting at 500 V. Monitor the Total Ion Count in the mass spectrometer software until ionization is detected.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Once the OptiMS sprayer position has been optimized based on the stability of the TIC, the minimum ESI voltage is determined. Increase the ESI spray voltage by approximately 200 V. This is your optimal ESI voltage.
CESI 8000 software. This sequence of events is very important because the mass spectrometer must be set up first so that it is waiting for the CESI 8000 Plus system to send the start signal, which will cause the mass spectrometer to User Manual...
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System start data acquisition. IMPORTANT Prior to attempting to run the beta-galactosidase sample, make sure the procedures for establishing a stable spray (refer to Establishing a Stable Spray and Determining Optimum ESI Voltage) and tuning (refer to Tuning) have been followed and successfully completed.
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Figure 9.12 First Segment of a Method Used to Run Beta-Galactosidase Digest Test Sample The tune method for the segment shown in Figure 9.13 is set with optimized ESI voltage for a stable spray.
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Figure 9.13 Second Segment of a Method Used to Run Beta-Galactosidase Digest Test Sample In the scan event settings, as illustrated in Figure 9.14, set the Data type...
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Create a Sequence with the Xcalibur Software The newly created beta-galactosidase mass spectrometer method should be programmed into a sequence. The sequence can be queued to start. The mass spectrometer will not begin to run the method until it receives a relay start trigger from the CESI 8000 software.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Click , and the sequence table dialog will appear as shown in Figure 9.16. Start a Sequence with the Xcalibur Software Figure 9.16 Sequence Setup-Home Page Highlight row #1 corresponding to the sample to run, and then click the toolbar icon.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Status tab, which is also on the left pane, will show the status of the sequence. As the sequence is downloaded and queued, the status will change from...
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Take the two buffer trays and place the vials with the reagents in the location as shown on Figure 9.17. Figure 9.17 Buffer Trays Inlet and Outlet for Beta-Galactosidase Separation Method Dispense the beta-galactosidase test sample into a micro vial, and place it inside of a CESI- MS vial.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Create a Sequence in the CESI 8000 Software In the CESI 8000 Plus system, the sequence to run beta-galactosidase will be performed using a method with initial conditions and time program as illustrated in Figure 9.19...
20 kV. Automatic Rinse Feature The CESI 8000 Plus system makes use of an “Automatic Rinse” feature. If there is a hardware failure during the rinse steps and also the injection step, either during a method run or sequence run, the instrument will (without user intervention) rinse the separation capillary for 3 minutes at 50 psi from the first separation event vial position.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Start a Sequence in the CESI 8000 Software Prior to starting a sequence, make sure that all reagents and sample required to run the experiment are in their appropriate place.
Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Click Start Beta-Galactosidase Analysis This section provides an example of beta-galactosidase analysis. From all 150 peptide fragments of beta-galactosidase detected, the 729 and 714 m/z will be used to evaluate the system performance.
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System Figure 9.24 Example of a Typical Beta-Galactosidase Total Ion Chromatogram Change the view from TIC (total ion chromatogram) to Base Peak Chromatogram. Configure the Base Peak Chromatogram in order to view the following two peaks: 714 and 729.
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Using a Thermo Scientific Mass Spectrometer with the CESI 8000 Plus System IMPORTANT The peak range 714.5 to 714.7 and 729.2 to 729.4 may need to be adjusted based on the mass spectrometer mass calibration. Figure 9.25 Identification and Clear Separation of the Beta-Galactosidase 714 and 729 m/z...
Cartridge Removal and Instrument Shutdown To remove the OptiMS cartridge from the CESI 8000 Plus system, make sure that the MS ESI voltage is off and then perform the following steps: If the cartridge is not going to be used for a short period of time (up to three days), then run the capillary cleaning method for short term storage.
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Figure 10.1 Load Button on Direct Control Window Open the cartridge cover (Figure 4.7). The CESI 8000 Plus coolant pump will start up and expel the coolant from the cartridge coolant lines (Figure 10.2). This will take about 30 seconds. Wait for the pump to turn off before removing the cartridge.
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To retract the stage for a Thermo Scientific mass spectrometer, refer to Aligning the CESI 8000 Plus System with a Thermo Scientific Mass Spectrometer CHAPTER To retract the stage for a SCIEX mass spectrometer, refer to Aligning the CESI 8000 Plus System with the ® SCIEX TripleTOF...
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Figure 10.5 OptiMS Capillary Sprayer Removal Insert the protective sleeve onto the OptiMS sprayer to protect it from breakage (Figure 10.6). Figure 10.6 Placing OptiMS Plastic Sleeve on Sprayer Tip CESI 8000 Plus High Performance Separation-ESI Module User Manual 220 / 376 RUO-IDV-05-3897-B | B11949AF...
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Pass the tubing with sprayer end through the access panel, and then remove coolant tubing from the notched arm (Figure 10.7 Figure 10.8). Figure 10.7 Putting Tubing/Sprayer through Access Panel Figure 10.8 Removing Coolant Tubing from Notch User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 221 / 376...
Decoupling the CESI 8000 Plus System If the mass spectrometer is to be used for a period of time without the CESI 8000 Plus system, or if one or both instruments are to be moved, then it is best to decouple the CESI 8000 Plus system from the mass spectrometer.
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Shutdown Procedures and Decoupling Instruments Shut down the CESI 8000 Plus system, and then unplug the power cord for the CESI 8000 Plus system and CESI 8000 Plus cart. Decouple both the mass spectrometer adapter’s high-voltage cables from the output and input...
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Shutdown Procedures and Decoupling Instruments IMPORTANT Always lower the CESI 8000 Plus cart to its lowest setting before moving the CESI 8000 Plus system. Figure 10.11 Unlocking CESI 8000 Plus Mobile Cart Wheels Use the thumbscrew on the Thermo Scientific mass spectrometer to loosen the adapter. Then remove the...
Cartridge Removal and Instrument Shutdown section earlier in this chapter. Shut down the CESI 8000 Plus system, and then unplug the power cord for the CESI 8000 Plus system and CESI 8000 Plus cart. User Manual...
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Disconnect the black and red ground banana plugs from the CESI 8000 Plus system panel. Unlock the CESI 8000 Plus cart wheels by lifting the lever at each caster upwards with your foot (Figure 10.15). Lower the cart to its lowest setting. Then slowly move the CE instrument away from the mass spectrometer.
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Shutdown Procedures and Decoupling Instruments Figure 10.15 Unlocking CESI 8000 Plus Mobile Cart Wheels Loosen the mass spectrometer adapter locking screw and then slide the adapter all the way off of the stage (Figure 10.16). Put the mass spectrometer adapter in a safe place.
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Shutdown Procedures and Decoupling Instruments CESI 8000 Plus High Performance Separation-ESI Module User Manual 228 / 376 RUO-IDV-05-3897-B | B11949AF...
Temperature: 15 °C to 40 °C (15 °C to 30° C recommended) Maximum Heat Dissipation 400 W (1024 BTU/hour) Pollution Degree TTL: 2 Contact closures: 2 User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 229 / 376...
• 2 serial ports • 2 Ethernet ports • 8 USB ports NOTE SCIEX fully validates and supports the controllers supplied with the system. Only limited support is available for customer-supplied computers. NOTE Specifications are subject to change without notice.
±0.3 psi at 25 psi ±1.0 psi at 100 psi Pressure Direction Applied at inlet or outlet for all pressure functions, rinses, and injections User-settable in software User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 231 / 376...
Output 2 not used Output 3 depends on what is programmed: Current signal when voltage is programmed Voltage signal when current or power is programmed CESI 8000 Plus High Performance Separation-ESI Module User Manual 232 / 376 RUO-IDV-05-3897-B | B11949AF...
UV Source 30 W pre-aligned deuterium lamp Scan Collection Frequency 0.5 Hz to 32 Hz Detector 256 element diode array Bandwith 6 nm minimum (absorbance averaging) User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 233 / 376...
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AU/V values can be set in the software. Output 3 depends on what is programmed: Current signal when voltage is programmed Voltage signal when current or power is programmed CESI 8000 Plus High Performance Separation-ESI Module User Manual 234 / 376 RUO-IDV-05-3897-B | B11949AF...
5600 mass spectrometer acquisition computer. One port can be used for another system (such as a SCIEX LC system), and one port can be dedicated to a CESI 8000 Plus system, and the third port is connected to the Internet.
(PC) used by the CESI 8000 Plus system. When reconnecting back to the CESI 8000 Plus system, you will want to reconnect to exactly the same network connector on the mass spectrometer acquisition computer that was used before switching.
Spectrometer Close the 32 Karat software on the CESI 8000 Plus controller. Disconnect the crossover network cable connecting the CESI 8000 Plus system to the mass spectrometer acquisition computer. Connect the relay/trigger cable from the CESI 8000 Plus system to the target mass spectrometer.
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B.4). Figure B.4 Change MS Type Dialog Click Change The CESI 8000 Plus system and the 32 Karat software are now properly configured for a non- SCIEX mass spectrometer. CESI 8000 Plus High Performance Separation-ESI Module User Manual 238 / 376...
Close the 32 Karat software on the CESI 8000 Plus controller. Disconnect the relay/trigger cable from the CESI 8000 Plus system. Reconnect the crossover network cable from the CESI 8000 Plus system to the SCIEX mass spectrometer acquisition computer. Verify the correct network settings by performing the steps in the...
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Switching to the CESI 8000 Plus System from Different Mass Spectrometer Systems section above within this appendix. Click Change The CESI 8000 Plus system and the 32 Karat software are now properly configured for a SCIEX mass spectrometer. CESI 8000 Plus High Performance Separation-ESI Module User Manual...
One port should be dedicated to another system (such as a SCIEX LC system), and one port should be dedicated to CESI 8000 Plus system. The third port connection is dedicated to the Internet. In the CESI 8000 Plus controller (PC), there are one (or more) network connections available.
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Nominal Network Configuration acquisition computer, and this connection will not change. That is, once connected, the network cable need never be removed from the CESI 8000 Plus controller. • The mass spectrometer acquisition computer has three network ports. • The CESI 8000 Plus controller has two available network ports.
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Leave the Internet Protocol TCP/IP Properties as shown (default) for normal Internet connection. Different IT environment can have different settings. Work with the local IT group to define those settings. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 243 / 376...
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Interface metric check box and enter (Figure B.13). Figure B.13 Advanced TCP/IP Settings 8. Close all windows and go back to the Network Connections page. CESI 8000 Plus High Performance Separation-ESI Module User Manual 244 / 376 RUO-IDV-05-3897-B | B11949AF...
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Figure B.14 Local Area Connection 2 10. Right-click the renamed network connection ( CESI 8000 ) and select Properties (Figure B.15). Figure B.15 CESI 8000 Plus Network Connection Properties 11. Highlight the Internet Protocol (TCP/IP) Properties and click (Figure B.16).
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Default gateway : Leave it blank. • Preferred DNS server : Do not change; leave it blank. Figure B.17 Internet Protocol (TCP/IP) Properties 14. Select Advanced CESI 8000 Plus High Performance Separation-ESI Module User Manual 246 / 376 RUO-IDV-05-3897-B | B11949AF...
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16. Click Network Connections and exit to the window. 17. Right-click the CESI 8000 network connection again and select Properties (Figure B.19). Figure B.19 Network Connections User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 247 / 376...
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20. Change Value from Auto Negotiation 100Mbps/Full Duplex (Figure B.21). Figure B.21 Link Speed and Duplex Settings 21. Click and exit all windows. CESI 8000 Plus High Performance Separation-ESI Module User Manual 248 / 376 RUO-IDV-05-3897-B | B11949AF...
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B.22). Figure B.22 Control Panel 2. Switch the Windows view to Classic View , if not already set (Figure B.23). Figure B.23 Selecting Classic View User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 249 / 376...
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8. Following the steps above, modify the network interface metric to Network Connection (or rename to other device interface name). CESI 8000 Plus High Performance Separation-ESI Module User Manual 250 / 376 RUO-IDV-05-3897-B | B11949AF...
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Figure B.26 Network and Sharing Center NOTE There will be more than one connection on the above window if the system has more than one network port installed. Locate an available network connection to set up the CESI 8000 Plus connection. 2. Click...
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Turn off file and printer sharing (Figure B.28). Figure B.28 Update Advanced Sharing Settings 4. Click Change adapter settings in the left pane (Figure B.29). Figure B.29 Change Adapter Settings CESI 8000 Plus High Performance Separation-ESI Module User Manual 252 / 376 RUO-IDV-05-3897-B | B11949AF...
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B.30). Figure B.30 Ethernet Properties 6. Highlight Internet Protocol Version 4 (TCP/IPv4) Properties and click (Figure B.31). Figure B.31 Internet Protocol Version 4 (TCP/IPv4) Properties User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 253 / 376...
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: Do not change; leave it blank. • Preferred DNS server : Do not change; leave it blank. Figure B.33 DNS Server Address 9. Click Advanced CESI 8000 Plus High Performance Separation-ESI Module User Manual 254 / 376 RUO-IDV-05-3897-B | B11949AF...
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13. Click Configure , which is located just below and to the right of Connect using (Figure B.35). Figure B.35 Configuring the Network User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 255 / 376...
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Panel Control Items > Windows Firewall > Customize Settings (Figure B.37). Select Turn off Windows Firewall in both Private and Public network settings. Click Figure B.37 Firewall Settings CESI 8000 Plus High Performance Separation-ESI Module User Manual 256 / 376 RUO-IDV-05-3897-B | B11949AF...
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Figure B.39 Network and Sharing Center NOTE There will be more than one connection at the above window if the system has more than one network port installed. Locate an available network connection to set up the CESI 8000 Plus connection. User Manual...
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Local Area Connection and select Properties (Figure B.41). Figure B.41 Local Area Connection Properties 7. Highlight Internet Protocol Version 4 (TCP/IPv4) and click Properties (Figure B.42). CESI 8000 Plus High Performance Separation-ESI Module User Manual 258 / 376 RUO-IDV-05-3897-B | B11949AF...
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Figure B.42 CESI 8000 (Local Area Connection) Properties Dialog 8. Click Use the following IP address (Figure B.43). Figure B.43 Internet Protocol Version 4 (TCP/IPv4) Properties User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 259 / 376...
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13. Right-click CESI 8000 and select Properties 14. Click Configure , which is located just below and to the right of Connect using (Figure B.45). CESI 8000 Plus High Performance Separation-ESI Module User Manual 260 / 376 RUO-IDV-05-3897-B | B11949AF...
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B.46). Figure B.46 Value List 16. Exit all network configuration windows. 17. Perform the steps in the “Network Sharing Setting for the Crossover Connection” section. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 261 / 376...
Navigate to the Network and Sharing Center and select options (Figure B.47). Figure B.47 Network and Sharing Center Select Change advanced sharing settings (Figure B.48). Figure B.48 Homegroup Options CESI 8000 Plus High Performance Separation-ESI Module User Manual 262 / 376 RUO-IDV-05-3897-B | B11949AF...
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B.49). Leave everything else as the default selection. Figure B.49 Advanced Sharing Settings Save changes Select to save all changes (Figure B.50). Figure B.50 Saving Changes User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 263 / 376...
Select the Connect to AB SCIEX MS check box, and verify/set the IP address 192.168.1.1 Select Change (Figure B.52). Figure B.52 Changing the MS Type CESI 8000 Plus High Performance Separation-ESI Module User Manual 264 / 376 RUO-IDV-05-3897-B | B11949AF...
Before installing the patch, open the hardware manager and deactivate all profiles. Make sure the ® Analyst TF software is closed before installing the patch. Locate the patch download file on the CESI 8000 Plus Sharepoint site. Unzip the file onto the desktop. .msi Double-click the...
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Nominal Network Configuration Figure B.55 License Agreement Figure B.56 Program Installation Figure B.57 Completion Wizard CESI 8000 Plus High Performance Separation-ESI Module User Manual 266 / 376 RUO-IDV-05-3897-B | B11949AF...
B.60, waiting for the wizard shown in Figure B.62 to complete, Finish and then selecting Figure B.64. Figure B.59 BCI CE-MS Driver — InstallShield Wizard User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 267 / 376...
— File location: C:\Program Files\BCI CE-MS Driver NOTE Refer to Figure B.63 for an example of a network configuration file. Figure B.63 Network Configuration File — Example User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 269 / 376...
To configure the Mass Spectrometer Hardware Profile ® Open the Hardware Configuration editor in the Analyst TF software (Figure B.64). Figure B.64 Hardware Configuration Editor CESI 8000 Plus High Performance Separation-ESI Module User Manual 270 / 376 RUO-IDV-05-3897-B | B11949AF...
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New Profile (Figure B.65). Figure B.65 Creating a New Profile Profile Name Name the profile by entering a name, such as “CESI 8000 Plus profile”, into the field (Figure B.66), and then selecting Figure B.66 Naming the Profile Add Device...
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Click Add Device Click again. Select Software Application from the Device Type menu (Figure B.68). Figure B.68 Device Type Menu in the Available Devices Dialog CESI 8000 Plus High Performance Separation-ESI Module User Manual 272 / 376 RUO-IDV-05-3897-B | B11949AF...
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Set up the device: a. From the Create New Hardware Profile window, click the mass spectrometer and select Setup Device (Figure B.70). Figure B.70 Added Devices User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 273 / 376...
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Figure B.72 Software Application Settings Make sure the BCI CE Driver is selected, and click Create New Hardware Profile From the window, click the Autosampler. a. Select Setup Device CESI 8000 Plus High Performance Separation-ESI Module User Manual 274 / 376 RUO-IDV-05-3897-B | B11949AF...
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Close all settings windows and navigate back to the dialog. Verify that the final hardware configuration profile appears (Figure B.74). Figure B.74 Final Hardware Configuration Profile User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 275 / 376...
(Figure B.75). Figure B.75 Hardware Configuration Editor Dialog Restart the SCIEX acquisition computer. Once the profile has been activated, the right corner of the Analyst window should appear as shown in Figure B.76.
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Figure B.78 Enter Password Dialog Adjust Curtain Gas Settings Set the curtain gas to in the window and click (Figure B.79). Figure B.79 Adjust Curtain Gas Setting Window User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 277 / 376...
Repeat the process until the read back value (see Curtain Gas Verification) is between Curtain Gas Verification To verify and adjust the curtain gas: Power on the CESI 8000 Plus system. ® In the Analyst TF software, double-click Manual Tuning...
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(Figure B.82). NOTE The Curtain Gas CUR field cannot be changed while the CESI 8000 Plus hardware profile is active. Figure B.82 Checking the Curtain Gas Readback Value User Manual CESI 8000 Plus High Performance Separation-ESI Module...
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Nominal Network Configuration NOTE As long as the user is running the CESI 8000 Plus hardware profile, the instrument pulls its curtain gas value from the curtain gas patch, rather than the SW input. Therefore, the Curtain ® TF software will not reflect the curtain gas setting of 5, nor can it...
TripleTOF 5600 Mass Spectrometer Manual and Auto-Calibration Methods Overview This appendix outlines the methods for manual and automatic calibration of the SCIEX ® TripleTOF 5600 mass spectrometer while performing CE-MS runs. When performing CESI-MS runs, the mass spectrometer can be calibrated periodically by infusing a calibrant solution through the OptiMS capillary and using it to automatically calibrate the mass spectrometer.
> > (Figure C.1). The Tuning Options dialog will open (Figure C.2). Figure C.1 Create Reference File Figure C.2 Tuning Options Dialog 1. Reference Button CESI 8000 Plus High Performance Separation-ESI Module User Manual 282 / 376 RUO-IDV-05-3897-B | B11949AF...
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C.5. Ensure that the radio button for Positive (“Positive Ion” mode) is checked and the Retention time tolerance +/- is set to 30 000 seconds. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 283 / 376...
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+/- 30 seconds. to open the Tuning Options window containing the newly created reference table Click (Figure C.6). Figure C.6 Tuning Options Dialog CESI 8000 Plus High Performance Separation-ESI Module User Manual 284 / 376 RUO-IDV-05-3897-B | B11949AF...
20 minutes by clicking Voltage . Ensure that the capillary inlet is in the BI:A1 position and the outlet is in the BO:A1 position (Figure C.8). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 285 / 376...
Under Period , set the Duration minute. Figure C.9 shows the MS tab in the Tune Method Editor dialog after setting up all the parameters. CESI 8000 Plus High Performance Separation-ESI Module User Manual 286 / 376 RUO-IDV-05-3897-B | B11949AF...
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(spray voltage) and adjust the sprayer position until the spray is stable. Start Mass Click , a TIC window will be acquired in approximately 1 minute. The Spectrum windows are shown in Figure C.11. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 287 / 376...
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C.12. Figure C.12 Opening a File from the TIC Window Pane of the data file that was acquired will open as shown in Figure C.13. CESI 8000 Plus High Performance Separation-ESI Module User Manual 288 / 376 RUO-IDV-05-3897-B | B11949AF...
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(Figure C.14). Figure C.14 Deleting MS Data Pane After deleting the pane, the will look as shown in Figure C.15. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 289 / 376...
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If the reference file B-Gal CE-MS Calibration Ref is not found, then refer to Create a Reference Table for CE-MS Calibration for the procedure on how to create this file. CESI 8000 Plus High Performance Separation-ESI Module User Manual 290 / 376 RUO-IDV-05-3897-B | B11949AF...
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Entire file In the area, click the icon to apply this calibration to all samples in the file. Click Save Figure C.18 TOC Calibration Toolbar User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 291 / 376...
Collision Energy Spread High Sensitivity tab, set . Ensure that Tune mode is selected. Figure C.20 shows the window after setting up all the parameters. CESI 8000 Plus High Performance Separation-ESI Module User Manual 292 / 376 RUO-IDV-05-3897-B | B11949AF...
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Product Ion calibration are shown in the window (Figure C.21). Figure C.21 TIC and Mass Spectrum Windows Open File Right-click the pane and click (Figure C.22). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 293 / 376...
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To open the average MS, click and drag to highlight the pane. Double-click it to obtain an average mass spectrum. Right-click the mass spectrum and click Re-calibrate TOF (Figure C.24). CESI 8000 Plus High Performance Separation-ESI Module User Manual 294 / 376 RUO-IDV-05-3897-B | B11949AF...
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, which is the same as the one used before for TOF Tolerance mass spectrometer calibration. Set the Da. The Product ion masses will appear in the table (Figure C.25). Figure C.25 TOF Calibration Window User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 295 / 376...
This method uses BGE vials that are to be placed in the A4 position of both the inlet and the outlet buffer trays. Figure C.26 Time Program CESI 8000 Plus High Performance Separation-ESI Module User Manual 296 / 376 RUO-IDV-05-3897-B | B11949AF...
In the pane, click In the tab, click Edit Parameters (Figure C.28). This will open a new window (Figure C.29) containing 2 tabs: Source/Gas Compound User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 297 / 376...
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Product Ion (+) 729.4 . In the tab, click Edit Parameters Source/Gas Compound to set up the parameters under for the MS/MS experiment (Figure C.31). CESI 8000 Plus High Performance Separation-ESI Module User Manual 298 / 376 RUO-IDV-05-3897-B | B11949AF...
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1. Product Ion (+) 729.4 Figure C.32 MS Tab and Parameters Settings Dialog Figure C.33 Acquisition Method and the Beckman CE Driver 1. Beckman CE Driver User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 299 / 376...
This section provides step-by-step instructions on how to build a sequence of 10 runs of beta- ® galactosidase performance test on theTripleTOF 5600 mass spectrometer with auto-calibration every 5 runs in tandem using the CESI 8000 Plus system. In the navigation bar, under Acquire, double-click Build Acquisition batch CESI 8000 Plus High Performance Separation-ESI Module...
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, browse to the applicable method. In this example choose . Refer to CESI_MS_AB_Sciex_Beta-Gal_45min CHAPTER Methods for the CESI 8000 Plus System for steps to create this method. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 301 / 376...
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“CE infusion” mode, all the reference ions are present at the same time. The tolerance is set to +/- 30 seconds. Reference tables are shown in Figure C.38. CESI 8000 Plus High Performance Separation-ESI Module User Manual 302 / 376 RUO-IDV-05-3897-B | B11949AF...
Start a Sequence in the Analyst TF Software Submit Click on the right to submit the batch. This will submit the 10 beta-galactosidase separation runs with auto-calibration (Figure C.39). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 303 / 376...
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At this point the TripleTOF 5600 mass spectrometer is waiting for the trigger signal from the CESI 8000 software to start data acquisition. CESI 8000 Plus High Performance Separation-ESI Module User Manual 304 / 376 RUO-IDV-05-3897-B | B11949AF...
A1 and A2 respectively. Samples to be analyzed can be placed in other positions; however, these positions must be specified in the CESI 8000 Plus methods or manually specified in the sequence table.
In the Method field, click the yellow folder to browse to the CESI method for the separation of beta-galactosidase (Figure 8.8), as shown in Figure C.43. CESI 8000 Plus High Performance Separation-ESI Module User Manual 306 / 376 RUO-IDV-05-3897-B | B11949AF...
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Data file name (Figure C.44). Figure C.44 Setting up the Sequence Wizard — Unknowns Page User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 307 / 376...
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To edit the sequence table in the CESI 8000 Insert Line software: highlight the first run, right-click the highlighted run, and click (Figure C.46). CESI 8000 Plus High Performance Separation-ESI Module User Manual 308 / 376 RUO-IDV-05-3897-B | B11949AF...
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. In the Sequence Properties dialog (Figure C.48), ensure that under File paths, the data is being saved on a desired folder. A folder can also be created. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 309 / 376...
Start a Sequence in the CESI 8000 Software To run this sequence, click the Run Sequence icon and the Run Sequence dialog opens. Click Start to initiate the sequence. CESI 8000 Plus High Performance Separation-ESI Module User Manual 310 / 376 RUO-IDV-05-3897-B | B11949AF...
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Once the CE method starts, the status in the MS Queue Server will change from PreRun NOTE ® When the trigger signal is sent from the CESI 8000 Plus system to the TripleTOF 5600 mass spectrometer, the Queue Server will change from PreRun to Acquiring. User Manual...
20 kV. Automatic Rinse Feature The CESI 8000 Plus system makes use of an “Automatic Rinse” feature. If there is a hardware failure during the rinse steps and also the injection step, either during a method run or sequence run, the instrument will (without user intervention) rinse the separation capillary for 3 minutes at 50 psi from the first separation event vial position.
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To get the list of ions present in the mass spectrum, right-click the mass spectrum and select List data MS before calibration . This is shown for the Figure C.52. Figure C.52 Obtaining List Data User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 313 / 376...
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Use as reference . This is the same reference table previously used for auto-calibration. This is shown in Figure C.54 for the MS before calibration CESI 8000 Plus High Performance Separation-ESI Module User Manual 314 / 376 RUO-IDV-05-3897-B | B11949AF...
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1. Mass Spectrometer Before Calibration 2. Data Table From Mass Spectrometer Before Calibration 3. Mass Spectrometer After Calibration 4. Data Table From Mass Spectrometer After Calibration User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 315 / 376...
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For evaluating the MS/MS mass accuracy, repeat steps MS/MS before calibration MS/MS after calibration For the MS/MS experiment, the mass shift after calibration is lower than it was before calibration (Figure C.57). CESI 8000 Plus High Performance Separation-ESI Module User Manual 316 / 376 RUO-IDV-05-3897-B | B11949AF...
Refer to the AB SCIEX Mass Calibration Tutorial found under Start Menu > All Programs > AB SCIEX > Analyst TF 1.7 > Hardware and Software Guides for more information. Successful auto-calibration runs have a green check mark. An open red circle with a diagonal line through it indicates that the sample was acquired but the calibration failed due to one or more of the reference ions failing to meet the calibration criteria.
If the mass shift is greater than or equal to 80% of the reference ions is over 100 ppm, manual calibration has to be performed. Additional troubleshooting can be found in Auto-calibration Troubleshooting Table. CESI 8000 Plus High Performance Separation-ESI Module User Manual 318 / 376 RUO-IDV-05-3897-B | B11949AF...
2. If manual calibration doesn’t help, instrument optimization has experiment. the MS/MS is less than to be performed. 100 ppm for ≥ 80% of the reference ions. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 319 / 376...
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® TripleTOF 5600 Mass Spectrometer Manual and Auto-Calibration Methods CESI 8000 Plus High Performance Separation-ESI Module User Manual 320 / 376 RUO-IDV-05-3897-B | B11949AF...
• Check both Inlet Outlet to increment the rinse vials at inlet and outlet buffer trays. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 321 / 376...
To set an injection step with auto vial increment, open the method on 32 Karat software and then open the injection step. The auto vial increment section is indicated on the figure below by a red square. CESI 8000 Plus High Performance Separation-ESI Module User Manual 322 / 376...
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12 repetitions of the same method. The table below summarizes which vial position will be used at each cycle during the above injection step. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 323 / 376...
To set a separation step with use auto vial increment, open the method on 32 Karat software and then open the separation step. The auto vial increment section is indicated on the figure below by a red square. CESI 8000 Plus High Performance Separation-ESI Module User Manual 324 / 376...
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12 cycles, which is equivalent to 12 repetitions of the same method. The table below summarizes which buffer vial positions will be used at each cycle during the above separation step. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 325 / 376...
6 Out vial inc 3 means that only the outlet vial will be incremented cycles. In another example, after 3 cycles. Figure D.8 Time Program CESI 8000 Plus High Performance Separation-ESI Module User Manual 326 / 376 RUO-IDV-05-3897-B | B11949AF...
In other words, the cycle count will re-start at 1. • If the sequence has different methods, the CESI 8000 Plus system will revert to the initial set of specified at the start of each method. Therefore, do not combine different methods if the auto vial increment feature is been used.
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Auto Vial Increment Feature CESI 8000 Plus High Performance Separation-ESI Module User Manual 328 / 376 RUO-IDV-05-3897-B | B11949AF...
Figure E.1 Profile of Phosphate Contamination NOTE During this method, the sprayer must rest in the holster and remain immersed in no more than 10 mL of DDI water. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 329 / 376...
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Conductive Liquid Capillary Conditioning To clean up phosphate contamination, set up the CESI 8000 Plus method (Figure E.2). Figure E.2 CLC Conditioning Method IMPORTANT This method uses reverse rinses. Only one vial is required in the buffer inlet tray, at position A1.
Replacing Fuses IMPORTANT To achieve optimal instrument performance, consistent user maintenance must be performed regularly on the CESI 8000 Plus system. A yearly planned maintenance by a SCIEX FSE is recommended. User Manual CESI 8000 Plus High Performance Separation-ESI Module...
Figure F.1 CESI-MS Vials 1. CESI-MS Vial Cap 2. Maximum Fill Level 3. CESI-MS Vials Put the cap on the vial and press it into position. CESI 8000 Plus High Performance Separation-ESI Module User Manual 332 / 376 RUO-IDV-05-3897-B | B11949AF...
4. Micro Vial inside CESI-MS Vial Place the micro vial in the CESI-MS vial. Put the cap on the CESI-MS vial. Put the capped vial in the tray. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 333 / 376...
CESI 8000 Plus system. WARNING Maintenance or repair procedures not specifically described in this manual present a risk of electrical shock or injury. Refer additional servicing to SCIEX Technical Support. WARNING Do not attempt to defeat any of the instrument interlocks or safety mechanisms.
Prepare the reagents listed above for the capillary shutdown by placing the reagent vials on the inlet and outlet buffer trays as indicated in Figure F.5. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 335 / 376...
If there is a spill, then it may become necessay to remove the OptiMS interface plate to clean the CESI 8000 Plus system. To remove and then re-install the OptiMS interface plate into a CESI 8000 Plus system, perform the following steps: Removing OptiMS Interface Plate Remove the OptiMS cartridge.
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Re-installing OptiMS Interface Plate Hold the handle of the OptiMS interface plate and insert it as shown in Figure F.8. Figure F.8 OptiMS Interface Plate Insertion User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 337 / 376...
Loosen the two thumbscrews and lift the cartridge lock down bar. Remove the OptiMS cartridge from the interface block. If the trays are still in the way, remove them for access to both opening levers. CESI 8000 Plus High Performance Separation-ESI Module User Manual 338 / 376...
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With your fingers on the sides of the lever, press the lever up into the interface block until it snaps into place. Repeat for the second lever. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 339 / 376...
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Pry gently with the electrode tool to remove the electrode from the interface block. e. Remove the electrode from the electrode tool. Repeat for second electrode. Install an electrode into the interface block. CESI 8000 Plus High Performance Separation-ESI Module User Manual 340 / 376...
Perform steps under Remove the electrodes one at a time as shown in Figure F.11. Immerse both electrodes in the DDI water with the opening levers. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 341 / 376...
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Lower the clamp bar and tighten the two thumbscrews. Close the cartridge cover. IMPORTANT To avoid corrosion of the ion source, frequently clean the dry chemical waste that accumulates beneath the sprayer. CESI 8000 Plus High Performance Separation-ESI Module User Manual 342 / 376 RUO-IDV-05-3897-B | B11949AF...
The instrument power should be on with the OptiMS cartridge installed. Slowly fill the coolant supply until the fill indicator is within the yellow lines in the coolant sight glass (Figure F.15). User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 343 / 376...
Using Vials and Hardware Maintenance Figure F.15 Coolant Sight Glass Location on the CESI 8000 Plus System Remove the coolant fill adapter and close the sample cover. The coolant fill adapter comes with a plunger. When adding coolant, the plunger should not be used.
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F.16). Use the tweezers that were supplied with the instrument (or your thumbnail and forefinger) to remove the quad rings. Figure F.16 Location of the Quad Rings on the CESI 8000 Plus System Install the new quad ring in the quad ring recess of the interface block.
Make sure to replace the fuses with the correct type and rating for continued protection against risk of fire and/or improper instrument operation. CAUTION If the fuses continue to blow after being replaced, contact a SCIEX FSE for additional assistance. Items required: •...
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8.0 A Slow Blow; ¼ inch (2 ea.) 6.3 A Time Delay; 20 mm (2 ea.) Reinstall the fuse block and reconnect the power cable. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 347 / 376...
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Using Vials and Hardware Maintenance CESI 8000 Plus High Performance Separation-ESI Module User Manual 348 / 376 RUO-IDV-05-3897-B | B11949AF...
IMPORTANT For any maintenance not covered in this manual, contact SCIEX Technical Support. Troubleshooting and Flow Diagrams The following is a list of possible problems when using the CESI 8000 Plus system with a mass spectrometer: NOTE Whenever the problem involves positioning the sprayer tip, make certain that the droplets fall freely from the tip and the sprayer tip does not touch the mass spectrometer inlet.
Instead of using the 32 Karat software button in this instance, use the Show 32 Karat button at the center bottom of the user interface dialog. CESI 8000 Plus High Performance Separation-ESI Module User Manual 350 / 376 RUO-IDV-05-3897-B | B11949AF...
Very gently submerge the sprayer end of the cartridge in the 50 mL tube (Figure G.2). NOTE When inserting sprayer end into tube, try to not touch the sides of the tube. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 351 / 376...
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Figure G.3 Wipe Clean Sprayer End with Dry Paper Wipe Reinsert the sprayer back into the mass spectrometer adapter. Remove the 50 mL tube from holster and empty contents. CESI 8000 Plus High Performance Separation-ESI Module User Manual 352 / 376...
Troubleshooting and Mobile Cart Homing Procedure Electrospray Cannot Be Established Between the CESI 8000 Plus System and Mass Spectrometer User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 353 / 376...
Troubleshooting and Mobile Cart Homing Procedure Electrospray is Detected at Zero Ion Spray (IS) Voltage while Applying CE Voltage CESI 8000 Plus High Performance Separation-ESI Module User Manual 354 / 376 RUO-IDV-05-3897-B | B11949AF...
Troubleshooting and Mobile Cart Homing Procedure Unstable Electrospray while Applying CE Voltage and IS Voltage User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 355 / 376...
Troubleshooting and Mobile Cart Homing Procedure Droplet at Capillary Tip Disappeared Indicating that Electrospray has Started, but No Mass Spectrometer Signal is Detected CESI 8000 Plus High Performance Separation-ESI Module User Manual 356 / 376 RUO-IDV-05-3897-B | B11949AF...
Troubleshooting and Mobile Cart Homing Procedure Signal of Total Ion Current (TIC) is Lower Than Usual and/or Background Signal Intensity Fluctuates More Than 40% User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 357 / 376...
Troubleshooting and Mobile Cart Homing Procedure Unstable CE Current CESI 8000 Plus High Performance Separation-ESI Module User Manual 358 / 376 RUO-IDV-05-3897-B | B11949AF...
Troubleshooting and Mobile Cart Homing Procedure No Analyte Signals Detected by Mass Spectrometer During a CE-MS Separation User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 359 / 376...
Troubleshooting and Mobile Cart Homing Procedure Some Analyte Peaks are Missing or Smaller Than Expected CESI 8000 Plus High Performance Separation-ESI Module User Manual 360 / 376 RUO-IDV-05-3897-B | B11949AF...
Troubleshooting and Mobile Cart Homing Procedure CE Current Gradually Decreases During CE-MS Separation User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 361 / 376...
Troubleshooting and Mobile Cart Homing Procedure The Iron-Acetate Cluster m/z 537 is Suppressing the Sample Signals CESI 8000 Plus High Performance Separation-ESI Module User Manual 362 / 376 RUO-IDV-05-3897-B | B11949AF...
Troubleshooting and Mobile Cart Homing Procedure High Voltage System Unable to Deliver the Required Current User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 363 / 376...
Figure G.4 Thermo Scientific Mass Spectrometer Sprayer Tip Distance and ESI Voltage Correlation Perform the following steps to optimize the sprayer tip position after placing OptiMS cartridge on CESI 8000 Plus system, the OptiMS sprayer on mass spectrometer adapter, and before performing a CESI-MS separation: Fill both OptiMS separation capillary and CLC with background electrolyte.
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Repeat this procedure: • After installing an OptiMS cartridge. • If signal CESI separation voltage is changed. • If a different background electrolyte is used. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 365 / 376...
Figure G.5 SCIEX Mass Spectrometer Sprayer Tip Distance and ESI Voltage Correlation Perform the following steps to optimize the sprayer tip position after placing OptiMS cartridge on the CESI 8000 Plus system, the OptiMS sprayer on mass spectrometer adapter, and before performing a CESI-MS separation: Repeat this procedure: Fill both OptiMS separation capillary and CLC with background electrolyte.
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Repeat this procedure: • After installing an OptiMS cartridge. • If signal CESI separation voltage is changed. • If a different background electrolyte is used. User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 367 / 376...
If the legs fail to function properly, unplug the power, wait 10 seconds, plug the power back in and repeat the above step. The above procedure must be performed anytime the power module is replaced. If this does not resolve the problem, then contact SCIEX Technical Support. CESI 8000 Plus High Performance Separation-ESI Module User Manual...
SCIEXUniversity SCIEX Support SCIEX and its representatives maintain a staff of fully-trained service and technical specialists located throughout the world. They can answer questions about the system or any technical issues that might arise. For more information, visit the SCIEX website at sciex.com...
Documentation for the hardware products can be found on the Customer Reference DVD that comes with the system or component. For the latest versions of the documentation, visit the SCIEX website at sciex.com. NOTE To request a free, printed version of this document, contact sciex.com/contact-us.
Conductive Liquid Capillary Conditioning, Beta-Galactosidase Stock Solution, 151, Configure Beta-Galactosidase Test Sample, 151, Application, Blocked Sprayer Tip, Configure Instrument, Buffer Tray Setup, 152, Consignes de sécurité avertissement, User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 371 / 376...
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Loading the Cartridge and Samples, Étiquette de mise en garde chariot mobile, Lock, Étiquette de mise en garde RoHS (Chine), Login, Étiquette de recyclage (WEEE), Logout, Étiquette marque CE, CESI 8000 Plus High Performance Separation-ESI Module User Manual 372 / 376 RUO-IDV-05-3897-B | B11949AF...
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Optional, Run Acquisition, Options Run Acquisition window, Graph, Run Application, Output Data File Name, 118, Run Complete, Output Data Path, 118, Run Number, Run Queue, User Manual CESI 8000 Plus High Performance Separation-ESI Module RUO-IDV-05-3897-B | B11949AF 373 / 376...
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