SCIEX CESI 8000 Plus User Manual
  1. Manuals
  2. Brands
  3. SCIEX Manuals
  4. Control Unit
  5. CESI 8000 Plus
  6. User manual

SCIEX CESI 8000 Plus User Manual

High performance separation-esi module
Hide thumbs Also See for CESI 8000 Plus:
Table of Contents

Advertisement

Quick Links

Download this manual
CESI 8000 Plus High Performance
Separation-ESI Module
User Guide
RUO-IDV-05-3897-E
December 2023

Advertisement

Table of Contents
loading
Need help?

Need help?

Do you have a question about the CESI 8000 Plus and is the answer not in the manual?

Questions and answers

Related Manuals for SCIEX CESI 8000 Plus

Summary of Contents for SCIEX CESI 8000 Plus

  • Page 1 CESI 8000 Plus High Performance Separation-ESI Module User Guide RUO-IDV-05-3897-E December 2023...
  • Page 2 SCIEX warranties are limited to those express warranties provided at the time of sale or license of its products and are the sole and exclusive representations, warranties, and obligations of SCIEX. SCIEX...

  • Page 3: Table Of Contents

    Syringe Pump........................22 High-Voltage Power Supply....................22 LED Indicators........................23 Cartridge and Sample Cover Interlocks................23 OptiMS Cartridge....................... 25 Mass Spectrometer Adapters.....................28 Required Reagents and Materials...................28 Customer-Supplied Equipment and Supplies..............29 User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 3/247...
  • Page 4 Manually Calibrate in Product Ion Mode................89 7 Run Workflow..........................94 Methods for the Silica Surface OptiMS Cartridge..............94 Initial Conditions Tab......................95 Time Program Tab......................95 Do a System Suitability Test....................97 CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 4/247...
  • Page 5 Conductive Liquid Capillary Contamination................151 Condition the Conductive Liquid Capillary............... 152 Autocalibration Troubleshooting....................153 No Electrospray Between CESI 8000 Plus System and Mass Spectrometer....... 155 Electrospray Is Detected When ESI Voltage Is Zero.............156 Electrospray Is Unstable When Voltage Is Applied............... 157 Droplet at Capillary Tip Disappeared But No Mass Spectrometer Signal Is Detected..158...
  • Page 6 A 32 Karat Software......................... 170 System Administration......................170 Controller and System Start-Up.................... 170 Controller and Network Login...................170 License Key........................171 Start the CESI 8000 Plus System..................171 Open the 32 Karat Software.....................171 Systems and Projects...................... 173 Online and Offline Modes....................173 Open the Direct Control Window...................174 Create a Toolbar Icon for the Direct Control Window............175...
  • Page 7 (Optional) PDA Detector Specifications................238 E Glossary of Symbols......................239 F Glossary of Warnings......................245 Contact Us..........................246 Customer Training.........................246 Online Learning Center......................246 Purchase Supplies and Reagents..................246 SCIEX Support........................246 Cybersecurity........................246 Documentation........................246 User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 7/247...

  • Page 8: Operational Precautions And Limitations

    WARNING! Warning identifies an action that can cause personal injury if precautions are not obeyed. CAUTION: Caution identifies an operation that can cause damage to the system or corruption or loss of data if precautions are not obeyed. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 9: Regulatory Compliance

    Equipment Directive 2012/19/EU, as implemented in EN 40519. Refer to the section: Waste Electrical and Electronic Equipment. • Packaging and Packaging Waste (PPW): Packaging and Packaging Waste Directive 94/62/EC User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 9/247...

  • Page 10: United States

    Removal of the covers is not required for routine maintenance, inspection, or adjustment. For repairs that require removal of the covers, contact a SCIEX field service employee (FSE). • Obey the required electrical safe work practices.

  • Page 11: Protective Earth Conductor

    For the health and safety precautions that must be obeyed for a chemical, refer to the safety data sheet (SDS). For storage information, refer to the certificate of analysis. To find a SCIEX SDS or certificate of analysis, go to sciex.com/tech-regulatory. • Always wear assigned personal protective equipment, including powder-free gloves, protective eyewear, and a laboratory coat.

  • Page 12: System Safe Fluids

    CAUTION: Potential System Damage. Do not use any other fluid until confirmation is received from SCIEX that it does not cause a hazard. This is not an exhaustive list. Any substance supplied with the CESI 8000 Plus system, or referenced in the documentation for the system, can safely be used with the system.

  • Page 13: Electromagnetic Environment

    This equipment generates, uses, and can radiate radio frequency energy and, if not installed and used in accordance with the operator's manual, can cause harmful interference to radio communications. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 13/247...

  • Page 14: Decommissioning And Disposal

    When the system is removed from service, obey national and local environmental regulations to divide and recycle different materials. Note: SCIEX will not accept any system returns without a completed Decontamination Form. Contact an FSE to get a copy of the form.

  • Page 15: Qualified Personnel

    Installation Qualification to help the customer become familiar with system operation, cleaning, and basic maintenance. If a system under warranty is serviced by personnel who are not authorized by SCIEX, then SCIEX is not responsible to repair any damage caused by the servicing.

  • Page 16: Performance Specifications

    WARNING! Personal Injury Hazard. Use SCIEX-recommended parts only. The use of parts that are not recommended by SCIEX or the use of parts for any purpose other than their intended purpose can put the user at risk of harm or have a negative effect on system performance.

  • Page 17: Introduction

    CE in an ultra low-flow format, which enhances the sensitivity of mass spectrometry while reducing ion suppression. In the CESI 8000 Plus system, sample components are separated by an electrical field that is applied between the inlet vial and the OptiMS sprayer. Conductive fluid is automatically delivered from a system vial to complete the CE circuit.
  • Page 18 Introduction Figure 2-1 CESI 8000 Plus System Conceptual Overview Item Description Cartridge Separation capillary Conductive liquid capillary Sprayer housing Adapter on the mass spectrometer Power supply for the mass spectrometer Current monitor for the CESI 8000 Plus system High-voltage power supply...

  • Page 19: Hardware Overview

    Introduction The following optional detectors are also available from SCIEX for use with the CESI 8000 Plus High Performance Separation-ESI Module: • An ultraviolet (UV) detector that allows for six different bandpass filters • A photo diode array (PDA) detector that is in the 190 nm to 600 nm range •...

  • Page 20: Sample-Handling System

    Introduction Figure 2-2 CESI 8000 Plus System Item Description Cartridge installed Access panel Sprayer High-voltage input connection from the mass spectrometer High-voltage output connection to the adapter on the mass spectrometer Holster for the sprayer Sample-Handling System WARNING! Personal Injury Hazard. When the sample cover is opened, wear protective eyewear, a lab coat, and gloves.
  • Page 21 CAUTION: Potential System Damage. To decrease the risk of breakage and expelled particles, use only SCIEX vials that are designed for the CESI 8000 Plus system. Examine every vial for damage before use. Do not use any vial that shows cracks or other damage.

  • Page 22: Syringe Pump

    48-vial sample tray Syringe Pump The CESI 8000 Plus system uses an internal pump mechanism to supply 0.1 psi to 25 psi to do pressure injections or low-pressure mobilizations. The pump can apply a maximum of 100 psi to move the fluids through the capillary. Pressure can also be applied to both ends of the cartridge (both capillaries) at the same time, so that both background electrolyte and conductive fluid can be supplied to the cartridge at the same time.

  • Page 23: Led Indicators

    The hinged doors of the CESI 8000 Plus system have interlock sensors that prevent unsafe access to the inside of the system. The outer door is the sample cover. The inner door is the cartridge cover.
  • Page 24 • If high voltage is on, then it turns off. • The pump that supplies the capillary coolant turns off. • The detector filter wheel moves to the closed position. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 25: Optims Cartridge

    The inlet ends of the capillaries are protected by sheaths that retract when the cartridge is installed in the CESI 8000 Plus system. When the cartridge is not installed in the system, locking mechanisms prevent the protective sheaths from retracting.
  • Page 26 Inlet end of separation capillary Outlet end of cartridge body The ESI needle, located in the sprayer tip, closes the circuit between the CESI 8000 Plus system and the mass spectrometer, allowing electrospray to occur. CESI 8000 Plus High Performance Separation-ESI...
  • Page 27 CESI-MS conditions. The sprayer locks into an adapter specifically designed to fit the NanoSpray ion source on the mass spectrometer. Figure 2-10 Adapter for the NanoSpray Ion Source User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 27/247...

  • Page 28: Mass Spectrometer Adapters

    • OptiMS Thermo MS Adapter for NanoSpray II, NanoSpray Flex, and NanoSpray Flex NG Ion Sources • OptiMS Bruker MS Adapter Instructions for the SCIEX adapter follow. For the others, read the Installation Guide for the mass spectrometer adapter in use. Required Reagents and Materials...

  • Page 29: Customer-Supplied Equipment And Supplies

    Introduction Table 2-2 Additional Supplies from SCIEX (continued) Material Part Number Auxiliary I/O Cable Assembly 5032115 GPIB Connector Extender with Green I/O Connector Kit A78960 CESI Vial (100 Pack) B11648 CESI Vial Cap (100 Pack) B24699 PCR Microvial (100 Pack)

  • Page 30: Set Up The Sytem

    CESI 8000 Plus system when the separation voltage is on. Install the Cartridge Before a cartridge can be installed, the interface plate must be installed on the CESI 8000 Plus system. Refer to the section: Remove and Install the Interface Plate.
  • Page 31 Set Up the Sytem Figure 3-2 Insertion Bar Open the access panel. Figure 3-3 Access Panel Put the sprayer housing through the side panel, and then put the sprayer in the holster. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 31/247...
  • Page 32 To install the cartridge body, put the ends of the cartridge on the aligning blocks of the interface plate at a 45-degree angle, and lightly push the cartridge in. Figure 3-5 Aligning Blocks CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 33 Figure 3-6 T Slider Item Description Align the T slider in the groove User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 33/247...
  • Page 34 Figure 3-7 Capillaries Below the Interface Block Item Description Capillaries can be seen below the interface block within the ejectors Put the coolant and capillary tubing through the notched arm. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 34/247...

  • Page 35: Connect The Gpib-Usb Cable

    The GPIB-USB cable connects the controller to the CESI 8000 Plus system. Make sure that the CESI 8000 Plus system controller (computer, monitor, keyboard, and mouse) is assembled. Make sure that both the CESI 8000 Plus system and the controller are connected to electrical power. User Guide...
  • Page 36 Connect the GPIB-USB cable to the port labeled “Controller” on the CESI 8000 Plus system. If required, then use a spacer. Connect the other end of the GPIB-USB cable to a USB port on the CESI 8000 Plus system controller.

  • Page 37: Connect An Auxiliary I/O Cable

    Set Up the Sytem Connect an Auxiliary I/O Cable The auxiliary input/output (I/O) cable connects the CESI 8000 Plus system to the mass spectrometer. Note: To use an Ethernet cable, refer to the section: Connect an Ethernet Cable. Install the white/black wire in pin position 9.

  • Page 38: Configure The Hardware Profile

    Set Up the Sytem Figure 3-12 Change MS Type Option Make sure that the Connect to AB SCIEX MS check box is cleared. Figure 3-13 Change MS Type Dialog Note: The IP address in the dialog should be the one that was preconfigured for the mass spectrometer.
  • Page 39 Figure 3-14 Navigation Bar Note: The acquisition computer on the mass spectrometer uses the Analyst TF software. For information, refer to the document: Analyst Software Advanced User Guide. Click New Profile. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 39/247...
  • Page 40 In the Profile Name field, type a name, and then click OK. Figure 3-16 Create New Hardware Profile Dialog Click Add Device. Select Mass Spectrometer TripleTOF 6600, and then click OK. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 41 In the Device Type list, select Software Application, and then click OK. Figure 3-18 Device Type: Software Application Click Add Device again. In the Device Type list, select Autosampler, and then click OK. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 41/247...
  • Page 42 10. To set up the mass spectrometer, do this: In the Create New Hardware Profile dialog, select Mass Spectrometer TripleTOF 6600, and then click Setup Device. Open the Communication tab. Accept the default settings, and click OK. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 42/247...
  • Page 43 11. To set up the software application, do this: In the Create New Hardware Profile dialog, select Software Application, and then click Setup Device. Make sure that the Beckman CE Driver is selected, and then click OK. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 43/247...
  • Page 44 12. To set up the autosampler, do this: In the Create New Hardware Profile dialog, select AutoSampler Agilent, and then click Setup Device. Open the Communication tab, and then click Advanced. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 45 The Hardware Configuration Editor dialog opens. 14. Select the profile that was just created, and then click Activate Profile. A green check mark shows next to the activated profile. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 45/247...

  • Page 46: Install The Adapter

    During the installation of the sprayer assembly, the glass tip of the sprayer is exposed. If the ion source stage is too close to the inlet on the mass spectrometer, then damage to the sprayer tip can occur. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 47 Figure 3-26 Orientation of the Adapter Before Installation Item Description High-voltage output cable Hook Adapter Ion source Make sure that the position of the adapter is tight, and then tighten the thumbscrew. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 47/247...
  • Page 48 During the installation of the sprayer assembly, if the ion source stage is too close to the inlet on the mass spectrometer, then damage to the sprayer tip can occur. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 49 Do not let the sprayer tip touch any surfaces. Figure 3-29 Install the Sprayer Assembly Item Description Sprayer assembly Unlock position Turn the sprayer assembly counterclockwise to lock it in the adapter. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 49/247...
  • Page 50 Set Up the Sytem Figure 3-30 Lock the Sprayer Assembly in the Adapter Item Description Sprayer in the Unlock position Sprayer in the Lock position CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 50/247...

  • Page 51: Connect The High-Voltage Cables

    Put the CESI 8000 Plus system approximately 4 inches (10 cm) away from the ion source stage. On the right side of the CESI 8000 Plus system, open the access door to show the high- voltage connections panel. Figure 3-31 High-Voltage Connections Panel...

  • Page 52: Align The System With The Mass Spectrometer

    If required, then use the coarse Z-axis adjustment knob to move the ion source stage and adapter fully away from the curtain gas plate. Make sure that the capillary will not touch the mass spectrometer when the stage moves forward. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 53 Ion source stage Note: Make sure that there is a cover on the source drain assembly on the mass spectrometer. The source drain assembly must be covered fully to operate correctly. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 53/247...
  • Page 54 If the ion source stage is too close to the inlet on the mass spectrometer, then damage to the sprayer tip can occur. Make sure that the CESI 8000 Plus system is approximately 4 inches (10 cm) away from the ion source stage so that the coolant tubes can be extended fully.
  • Page 55 Coolant tubes Ion source stage Mobile cart At the same time, align the sprayer tip with the height indicator on the CESI 8000 Plus system and the center of the inlet on the mass spectrometer. User Guide CESI 8000 Plus High Performance Separation-ESI...
  • Page 56 Use the Up and Down buttons on the front of the mobile cart to adjust the height of the cart until the height indicator on the CESI 8000 Plus system aligns with the sprayer tip. WARNING! Personal Injury Hazard. Remove consumables from the mobile cart before the height is adjusted.
  • Page 57 CESI 8000 Plus High Performance Separation-ESI Module User Guide. When the CESI 8000 Plus system is in visual alignment with the mass spectrometer, step on the locking lever of each caster to lock each wheel of the mobile cart in position.
  • Page 58 If the sprayer tip is too close to the inlet, then the accidental suction of rinse solution into the mass spectrometer can occur and cause damage. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 59 Figure 3-38 XYZ-Axes Adjustment Knobs Item Description Height indicator on the CESI 8000 Plus system Fine Z-axis adjustment knob (movement toward the curtain gas plate) Coarse Z-axis adjustment knob (movement toward the curtain gas plate) Y-axis adjustment knob (vertical movement)

  • Page 60: Prepare The Reagents

    Only prepare new 20% HAc solution when the 200 mM LE buffer, pH 4.0 is prepared. In a 100 mL glass bottle, add 80 mL of CE Grade Water. Inside a fume hood, add 20 mL of HAc to the bottle. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 61: Prepare The 200 Mm Le Buffer, Ph 4.0

    In a 0.5 mL centrifuge tube, add 50 µL of PepCalMix and 50 µL of 10% HAc solution. Mix the solution in a vortex mixer for 10 seconds. Put the sample vial in the inlet sample tray in position A1. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 61/247...

  • Page 62: Prepare The Peptide Test Sample

    Mix the solution in a vortex mixer for 3 seconds. Keep the sample at 2 °C to 8 °C. At the end of the day, discard the solution that was not used. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 63: Prepare The Trays

    Prepare the Trays Prepare the Trays Use only CESI vials and caps in the CESI 8000 Plus system buffer and sample trays. It is important for each buffer and sample to be in the location identified in the tray layout. The buffer and sample locations are related to the method in the 32 Karat software.

  • Page 64: Fill A Cesi Vial

    Note: When filling the vials, do not make air bubbles. If there are air bubbles in the vial, then spin the vial in a centrifuge for a few seconds to remove the bubbles. CESI 8000 Plus High Performance Separation-ESI User Guide...

  • Page 65: Fill A Microvial

    Note: When filling the vials, do not make air bubbles. If there are air bubbles in the vial, then spin the vial in a centrifuge for a few seconds to remove the bubbles. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module...

  • Page 66: Fill A Nanovial

    Put the sample vial in the inlet sample tray at position S1:A1. Note: Make sure that the tab on the outside of the nanoVial points to the front of the sample tray. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 67: Prepare The Buffer Trays

    Note: When filling the vials, do not make air bubbles. If there are air bubbles in the vial, then spin the vial in a centrifuge for a few seconds to remove the bubbles. Prepare the Buffer Trays Use the following table to prepare the required reagents. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 67/247...

  • Page 68: Prepare The Sample Tray

    Use the following figure to put each reagent vial in the correct position in the buffer inlet and outlet trays. Note: This layout is the same for all five methods in the CESI 8000 Plus software. Figure 5-5 Buffer Tray Layout Prepare the Sample Tray Sample can evaporate over time.
  • Page 69 Prepare the Trays Figure 5-6 Sample Tray Layout User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 69/247...

  • Page 70: Prepare The System

    Falcon tube. If there is more than 10 mL in the tube, then the liquid can splash onto the metal components of the sprayer and cause damage. Carefully immerse the sprayer tip in the Falcon tube. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 71 11. Install the sprayer in the adapter on the mass spectrometer. The cartridge is ready to set a stable spray. 12. Remove the Falcon tube from the holster, and discard the contents. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 71/247...

  • Page 72: Establish A Stable Spray

    Rinse Direction icon (a blowing face identified by a red arrow in the following figure). The reverse direction refers to rinsing the conductive liquid capillary, and the blowing face shows on the right side of the window. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 73 Figure 6-5 Droplet at the End of the Stainless Steel Needle When the rinse is complete, to fill the separation capillary with BGE, do this: User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 73/247...
  • Page 74 Click OK. When the separation capillary is filled with BGE, a droplet will show at the end of the sprayer tip. Figure 6-7 Droplet at the End of the Sprayer Tip CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 75: Optimize The Esi Voltage

    3 μA. If the BGE used is 50 mM AmAc buffer, pH 3.0, then the electrical current should be 0.5 µA to 1 µA. In the Analyst TF software, in the Navigation bar, under Tune and Calibrate, double-click Manual Tuning. On the Source/Gas tab, do this: User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 75/247...
  • Page 76 In the TOF Masses (Da) Max field, type 2000. In the Period Duration field, type 10. Click Start. If a non-SCIEX mass spectrometer is in use, then do this on the mass spectrometer acquisition computer: Set the ESI voltage to zero.
  • Page 77 In the Period Duration field, type 5. 17. Click Acquire, and then type a file name (for example, Baseline). 18. If a non-SCIEX mass spectrometer is in use, then do this on the mass spectrometer acquisition computer: Set the ESI voltage to 1.0 kV and continue to acquire data.
  • Page 78 Fine-Tune the Position of the Sprayer Tip. 20. If a non-SCIEX mass spectrometer is in use, then do this: Monitor the spray stability for 20 minutes to 30 minutes to make sure that the baseline fluctuation is ≤ 40%. If the baseline fluctuation is > 40%, then condition the capillaries. Refer to the section: Condition the Capillaries.

  • Page 79: Fine-Tune The Position Of The Sprayer Tip

    In the 32 Karat software, use the Direct Control window to apply 20 kV voltage. • Make sure that the cartridge does not spray when the ESI voltage is 0 V during mass spectrometer scanning. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 79/247...

  • Page 80: Manual Calibration

    Autocalibration. For more information about calibration, refer to the document: AB SCIEX Mass Calibration Tutorial. To open the document, click Start Menu > All Programs > AB SCIEX > Analyst TF 1.7 > Hardware and Software Guides. Prepare for Manual Calibration In a sample vial, add 90 μL of PepCalMix and 10% HAc solution to make a 1:1 solution.

  • Page 81: Create A Reference Table

    Make sure that the capillary inlet is in position BI:A1 and the outlet is in position BO:A1. Click OK. Create a Reference Table In the Analyst TF software, in the Navigation bar, click Acquire. Click Tools > Settings > Tuning Options, and then click Reference. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 81/247...
  • Page 82 Figure 6-15 New Reference Table in the Reference Table Editor Window In the Name field, type a name that identifies the calibration solution (in this example, PepCalMix). Make sure that Positive is clicked. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 83 TVESLFPEEAETPGSAVR 964. 97741 In the Reference Ions for MS/MS Calibration table, type the following values. Table 6-2 Reference Ions for MS/MS Calibration Fragment Name Fragment m/z (Da) 185.09207 348.15540 User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 83/247...
  • Page 84 Figure 6-16 Tuning Options Dialog 11. Make sure that the name of the new reference table (in this example, PepCalMix) shows in the Positive Reference field. 12. Click OK. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 84/247...

  • Page 85: Manually Calibrate In Tof Ms Mode

    In the TOF Masses (Da) Min field, type 350. In the TOF Masses (Da) Max field, type 1000. In the Period Duration field, type 1000. Open the Advanced MS tab. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 85/247...
  • Page 86 Figure 6-19 TIC and Mass Spectrum Windows To open windows that show the TIC and mass spectrum for the data file that was acquired, right-click the TIC window, and then click Open File. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 87 10. To delete the mass spectrum pane, click the pane, and then on the toolbar, click the Delete icon. 11. To get an average mass spectrum, click and drag the cursor to highlight the TIC window, and then double-click the TIC window. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 87/247...
  • Page 88 Prepare the System Figure 6-21 Average Mass Spectrum Pane 12. Right-click the mass spectrum pane, and then click Re-Calibrate TOF. Figure 6-22 TOF Calibration Window CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 88/247...

  • Page 89: Manually Calibrate In Product Ion Mode

    23. Close the TIC and mass spectrum panes. Manually Calibrate in Product Ion Mode In the Analyst TF software, in the Navigation bar, under Tune and Calibrate, double-click Manual Tuning. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 89/247...
  • Page 90 In the TOF Masses (Da) Max field, type 1500. Make sure that High Sensitivity is clicked. Click Start. Windows open to show the total ion chromatogram (TIC) and mass spectrum generated during the product ion calibration. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 90/247...
  • Page 91 TOF-MS and MS/MS calibration. Note: The spectrum that shows cannot be used for calibration. Click the mass spectrum window, and then the toolbar, click the Delete icon. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 91/247...
  • Page 92 To get an average mass spectrum, click and drag the cursor to highlight the TIC window, and then double-click the TIC window. Figure 6-26 Re-Calibrate TOF Right-click the mass spectrum window, and then click Re-Calibrate TOF. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 93 15. To apply this calibration to all samples in the file, click Entire File. 16. Click Save. 17. Click OK twice. 18. To close the TOF Calibration window, click Close. 19. Close the TIC and mass spectrum windows. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 93/247...

  • Page 94: Run Workflow

    To use the CESI 8000 Plus system with the mass spectrometer, first set up a sequence in the Analyst TF software and then in the 32 Karat software. The mass spectrometer must be set up before the CESI 8000 Plus system sends the start signal, so that the mass spectrometer can start data acquisition.

  • Page 95: Initial Conditions Tab

    All of the methods for the silica surface OptiMS cartridge use the initial conditions in the following figure. Figure 7-1 Initial Conditions Tab Time Program Tab Each of the methods for the silica surface OptiMS cartridge uses a different time program. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 95/247...
  • Page 96 Figure 7-3 Time Program for the Separation Method Note: For the system suitability test, do not use a separation voltage that is > 20 kV. Figure 7-4 Time Program for the Autocalibration Method CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 97: Do A System Suitability Test

    MS and MS/MS levels, do a system suitability test. In the Analyst TF software, in the Navigation bar, under Acquire, double-click Build Acquisition Method. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 97/247...
  • Page 98 • The value for Duration is at least 1.5 minutes shorter than the duration set in the method in the 32 Karat software. In the Acquisition method pane, click TOF MS (+). CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 99 Figure 7-9 TOF MS (+) Acquisition Method Window: MS Tab Click Edit Parameters. Figure 7-10 TOF MS(+) Parameter Settings Dialog: Source/Gas Tab Type the values that show in the example. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 99/247...
  • Page 100 Figure 7-11 TOF MS(+) Parameter Settings Dialog: Compound Tab Open the Advanced MS tab, and then type values for the other parameters. Figure 7-12 TOF MS(+) Acquisition Method Window: Advanced MS Tab CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 101 In the Acquisition method pane, click Product Ion (+). Figure 7-13 Product Ion Acquisition Method Window: MS Tab Click Edit Parameters. Figure 7-14 Product Ion Parameter Settings Dialog: Source/Gas Tab User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 101/247...

  • Page 102: Create A Sequence In The 32 Karat Software

    Figure 7-15 Product Ion Parameter Settings Dialog: Compound Tab 11. Save the method file with the name CESI_MS_Sciex_PepCalMix_45min. Create a Sequence in the 32 Karat Software In the 32 Karat software, click File > Sequence > Sequence Wizard. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 103 Figure 7-16 Sequence Wizard - Method Page To the right of the Method field, click , and then select the CESI-MS separation method. Click Next. Figure 7-17 Sequence Wizard - Unknowns Page User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 103/247...
  • Page 104 Sample ID field. When the sequence is run, the parameter is added to the data file name automatically. Note: The Open File option is used to reprocess data files, not to acquire data. Select the applicable options. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 105 Note: If the Advance check box is selected for the inlet, outlet, or both vials, then the values for the vial positions are filled down automatically in the Sample Inject Inlet/Outlet columns in the sequence table. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 105/247...

  • Page 106: Create A Batch In The Analyst Tf Software

    In the Analyst TF software, in the Navigation bar, under Acquire, double-click Build Acquisition batch. The Batch Editor window opens. Click Add Set. A new set called SET1 is added. Click Add Samples. The Add Sample dialog opens. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 106/247...
  • Page 107 In the Sub Folder field, type the data file name, or click Browse and then find the applicable data file. In the New samples Number field, type 10. Figure 7-22 Add Sample Dialog Click OK. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 107/247...
  • Page 108 11. Click Method Editor, and then find the applicable method file (in this example, CESI_MS_Sciex_PepCalMix_45min). 12. Click File > Save as, and then type a name. 13. In the Batch Editor window, open the Calibrate tab. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 109 Table 7-2 Reference Ions for TOF MS Calibration Compound Name Precursor m/z (Da) CE for DP for MS/MS MS/MS AETSELHTSLK 408.55010 GAYVEVTAK 473.26020 IGNEQGVSR 485.25302 LVGTPAEER 491.26559 LDSTSIPVAK 519.79969 AGLIVAEGVTK 533.32333 User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 109/247...
  • Page 110 ALENDIGVPSDATVK 768.90340 AVYFYAPQIPLYANK 883.47380 TVESLFPEEAETPGSAVR 964. 97741 Table 7-3 Reference Ions for MS/MS Calibration Fragment Name Fragment m/z (Da) 185.09207 348.15540 560.30788 661.35555 758.91050 799.44398 856.46544 957.51312 1070.59719 1169.66560 CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 110/247...

  • Page 111: Start The Batch In The Analyst Tf Software

    The Queue Manager window opens to show that there are 12 runs (10 PepCalMix separations and 2 autocalibrations) in the queue. The hourglass symbol in column 1 shows that the queue is in Standby mode. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 111/247...

  • Page 112: Load The Trays

    Put the PepCalMix test sample for autocalibration in position SI:A1 of the sample inlet tray. Put the PepCalMix test sample for the system suitability test in position SI:A2 of the sample inlet tray. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 113: Start The Sequence In The 32 Karat Software

    Put the inlet tray in the inlet carrier (left) and the outlet tray in the outlet carrier (right), and then close the front cover. Start the Sequence in the 32 Karat Software On the toolbar, click the Sequence Run icon User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 113/247...
  • Page 114 Acquired. To see the status of the first autocalibration run, monitor the PepCalMix_Installation sequence. The first autocalibration run in the following figure is being acquired. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 115: Analyze The Data From The Pepcalmix Test Sample

    In the Analyst TF software, open the PepCalMix wiff file. The following figure shows the total ion chromatograms (TICs) for the separation of PepCalMix on three different bare fused-silica (BFS) cartridges. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 115/247...
  • Page 116 739.35 m/z and 768.90 m/z of the PepCalMix test sample run. The difference in migration time for peaks 485.25 m/z and 491.25 m/z is 0.08 minutes. The difference in migration time for peaks 739.35 m/z and 768.90 m/z is 0.25 minutes. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 117: About Autocalibration

    Note: Do manual calibration before autocalibration is done. If the reference ions are outside of the 100 ppm tolerance for peak identification, then autocalibration will not be successful. Note: The following procedures are for the SCIEX 5600 and 6600 systems. For other SCIEX mass spectrometers or mass spectrometers from other manufacturers, use the manufacturer's recommendations.

  • Page 118: Create The Ms Acquisition Method For Autocalibration

    Create the MS Acquisition Method for Autocalibration Use this procedure when PepCalMix is the sample solution. In the Analyst TF software, in the Navigation bar, under Acquire, double-click Build Acquisition Method. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 119 In the Curtain Gas (CUR) field, type 10. In the IonSpray Voltage Floating (ISVF) field, type 1500. The ISVF parameter sets the ESI voltage. In the Interface Heater Temperature (IHT) field, type 50.0. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 119/247...
  • Page 120 Run Workflow Figure 7-35 TOF MS (+) Parameter Settings Dialog: Source/Gas Tab Open the Compound tab, and then do this: CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 120/247...
  • Page 121 In the Collision Energy (CE) field, type 10. In the Acquisition method pane, click Product Ion (+) 758.91. Figure 7-37 Acquisition method Pane: Product Ion On the MS tab, click Edit Parameters. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 121/247...
  • Page 122 In the IonSpray Voltage Floating (ISVF) field, type 1600. The ISVF parameter sets the ESI voltage. In the Interface Heater Temperature (IHT) field, type 50.0. Open the Compound tab, and then do this: CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 123: Add Autocalibration To The Sequence

    Click to highlight the first run, right-click the highlighted run, and then click Insert Line. At the bottom of the Method column, click the green arrow icon and then click CESI-MS Auto-calibration_ABSciex.met. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 123/247...
  • Page 124 Make sure that the File paths for Method and Data show the correct folder to save the data, and then click OK. Note: If required, then create a folder. Click File > Sequence > Save As. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 125: Analyze The Autocalibration Data

    To extract the mass spectra from the TIC, highlight the region and then double-click the region. Extract one mass spectrum between 2 minutes to 2.5 minutes (MS before calibration) and another between 4 minutes to 4.5 minutes (MS after calibration). User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 125/247...
  • Page 126 MS/MS after calibration To get a list of ions in the mass spectrum, right-click the mass spectrum and then click List data. Do this for both MS before calibration and MS after calibration. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 127 Figure 7-43 Obtaining List Data To show all the reference ions used for calibration, in each data list table, open the Calibration Peak List tab. Figure 7-44 Reference Ions User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 127/247...
  • Page 128 To make sure that the correct reference table has been selected, right-click the reference table, click the PepCalMix_CE-MS_Calibration Ref reference table, and then click Use as reference. Figure 7-45 Reference Table Verification for MS Before Calibration Figure 7-46 MS After Calibration Analysis CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 128/247...
  • Page 129 In the Calibration peak list table, if the value in the Mass shift (ppm) field is > 100 ppm, then do manual calibration. Refer to the section: Manual Calibration. Make sure that the threshold for peak detection is 1% in the spectrum. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 129/247...
  • Page 130 In the Navigation bar, under Tune and Calibrate, click Tools > Settings > Appearance Options > Other Graph. In the Default Threshold for the Spectrum field, type 1. For more autocalibration troubleshooting, refer to the section: Autocalibration Troubleshooting. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 130/247...

  • Page 131: Shutdown And Disconnection

    Shutdown and Disconnection Shutdown and Disconnection Remove the Cartridge If the CESI 8000 Plus system will be shut down and the power turned off, then remove the cartridge. Make sure that the ESI voltage on the mass spectrometer is off.
  • Page 132 Let the ion source cool for at least 30 minutes before the sprayer is removed from the adapter on the mass spectrometer. Figure 8-3 Unlock the Sprayer from the Cartridge Item Description Sprayer in the Unlock position CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 132/247...
  • Page 133 11. Put the tubing with the sprayer end through the access panel. Figure 8-5 Sprayer and Tubing Through the Access Panel 12. Remove the coolant tubing from the notched arm. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 133/247...
  • Page 134 Note: Drops of liquid coolant that might fall from the cartridge ends are normal and do not cause damage to the hardware. 14. Refer to the section: Store the Cartridge. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 134/247...

  • Page 135: Store The Cartridge

    Mass Spectrometer Disconnect the CESI 8000 Plus system from the mass spectrometer when: • The mass spectrometer will be used without the CESI 8000 Plus system. • The mass spectrometer or the CESI 8000 Plus system will be moved. Make sure that the cartridge has been removed.
  • Page 136 3-31. Adjust the height of the mobile cart to the lowest position. Disconnect the power cables from the CESI 8000 Plus system and the mobile cart. To unlock the wheels on the mobile cart, lift the lever at each caster.
  • Page 137 Shutdown and Disconnection Figure 8-9 Loosen the Thumbscrew User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 137/247...

  • Page 138: Maintenance

    WARNING! Electrical Shock Hazard. To prevent the risk of electrical shock or injury, do not do maintenance or repair procedures that are not included in this manual. Contact a SCIEX field service employee (FSE) for maintenance service and support. WARNING! Electrical Shock Hazard. Do not try to disable any of the system interlocks or safety mechanisms.
  • Page 139 To install the interface plate, do this: Hold the handle on the top of the interface plate and install it. Make sure that the interface plate is flush with the mounting surface of the CESI 8000 Plus system. User Guide...
  • Page 140 Maintenance Figure 9-2 Install the Interface Plate Tighten both thumbscrews. Figure 9-3 Tighten the Thumbscrews CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 140/247...

  • Page 141: Interface Block, Electrodes, And Insertion Levers

    Interface Block, Electrodes, and Insertion Levers Figure 9-4 Interface Block, Electrodes, and Insertion Levers Item Description Interface block Electrodes Insertion levers Replace the Insertion Levers Required Materials • Insertion Lever Interface Parts Kit User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 141/247...

  • Page 142: Replace The Electrodes

    Make sure that the nub at the end of the electrode tool handle goes into the notch on the interface block. Pry carefully with the electrode tool to remove the electrode from the interface block. Remove the electrode from the electrode tool. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 143 Push up to install the electrode in the interface block with a click. To remove the electrode tool, pull straight back. Install the insertion levers. Refer to the section: Replace the Insertion Levers. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 143/247...

  • Page 144: Clean The Electrodes, Insertion Levers, And Interface Block

    Remove the insertion levers. Refer to the section: Replace the Insertion Levers. Immerse both insertion levers in a beaker with at least 150 mL of CE Grade Water. Remove the electrodes. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 144/247...

  • Page 145: Add Capillary Cartridge Coolant

    CAUTION: Potential System Damage. To prevent damage, do not use the plunger in the coolant fill tool when coolant is added. Gravity supplies sufficient force to pull the coolant into the system. Open the sample cover. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 145/247...
  • Page 146 Figure 9-8 Coolant Fill Tool Connected to the Coolant Fill Port Fill the syringe with 120 mL of coolant. Make sure that the CESI 8000 Plus system is on and a cartridge is installed. Slowly add coolant until the fill indicator is between the yellow lines in the coolant sight glass.

  • Page 147: Replace The Quad Rings

    Open the cartridge cover. Loosen the two thumbscrews on the insertion bar, and then lift the insertion bar. Turn off the power to the CESI 8000 Plus system. Remove the interface plate. Refer to the section: Remove and Install the Interface Plate.

  • Page 148: Replace The Fuses

    • Replacement fuses (Qty: 2) WARNING! Fire Hazard. Make sure to replace the fuses with the correct type and rating for continued protection against the risk of fire and incorrect system operation. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 149 Disconnect the power cable from the AC power outlet. Use a flat-bladed screwdriver to remove the fuse block. Figure 9-11 Fuse Block Item Description Fuse block Air conditioning vents External connections panel Replace the fuses. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 149/247...

  • Page 150: Set The Home Position For The Mobile Cart

    If the legs do not operate correctly, then do this: Disconnect the power cable. Wait 10 seconds. Connect the power cable. Do step again. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 150/247...

  • Page 151: Troubleshooting

    If the baseline mass spectrum shows a series of peaks with an m/z difference of 98, then the conductive liquid capillary might be contaminated with phosphate. To remove phosphate contamination, condition the conductive liquid capillary. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 151/247...

  • Page 152: Condition The Conductive Liquid Capillary

    Use the following figure to put each vial in the correct position in the buffer inlet and outlet trays. Note: This method uses reverse rinses. Only one vial is required in the inlet buffer tray, at position A1. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 153: Autocalibration Troubleshooting

    Prepare a fresh PepCalMix sample for autocalibration. In the TOF MS calibration data, find the missing ion, and then delete that ion from the reference table. Run the sequence again. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 153/247...
  • Page 154 The mass shift of the In the autocalibration data reference ions is not correct. file, if the mass shift of the reference ions is > 100 ppm, then do manual calibration. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 154/247...

  • Page 155: No Electrospray Between Cesi 8000 Plus System And Mass Spectrometer

    Troubleshooting No Electrospray Between CESI 8000 Plus System and Mass Spectrometer User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 155/247...

  • Page 156: Electrospray Is Detected When Esi Voltage Is Zero

    Troubleshooting Electrospray Is Detected When ESI Voltage Is Zero CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 156/247...

  • Page 157: Electrospray Is Unstable When Voltage Is Applied

    Troubleshooting Electrospray Is Unstable When Voltage Is Applied User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 157/247...

  • Page 158: Droplet At Capillary Tip Disappeared But No Mass Spectrometer Signal Is Detected

    Troubleshooting Droplet at Capillary Tip Disappeared But No Mass Spectrometer Signal Is Detected CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 158/247...

  • Page 159: Tic Signal Is Lower Than Usual And/Or Intensity Of Background Signal Fluctuates Too Much

    Troubleshooting TIC Signal Is Lower than Usual and/or Intensity of Background Signal Fluctuates Too Much User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 159/247...

  • Page 160: Electrical Current Is Unstable

    Troubleshooting Electrical Current Is Unstable CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 160/247...

  • Page 161: Analyte Signals Are Not Detected By Mass Spectrometer During Separation

    Troubleshooting Analyte Signals Are Not Detected by Mass Spectrometer During Separation User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 161/247...

  • Page 162: Some Analyte Peaks Are Missing Or Smaller Than Expected

    Troubleshooting Some Analyte Peaks Are Missing or Smaller than Expected CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 162/247...

  • Page 163: Current Gradually Decreases During Separation

    Troubleshooting Current Gradually Decreases During Separation User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 163/247...

  • Page 164: Iron-Acetate Cluster M/Z 537 Is Suppressing Sample Signals

    Troubleshooting Iron-Acetate Cluster m/z 537 Is Suppressing Sample Signals CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 164/247...

  • Page 165: High-Voltage System Cannot Deliver Required Electrical Current

    Troubleshooting High-Voltage System Cannot Deliver Required Electrical Current User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 165/247...

  • Page 166: Order Parts

    • Internet: For customers in the United States, Canada, United Kingdom, Belgium, Netherlands, France, Germany, and Switzerland, go to store.sciex.com Cartridges and Parts Note: Parts that have an asterisk (*) after the part number are available only from a SCIEX sales representative. Table 11-1 Cartridges, Capillaries, and Apertures Part Number...
  • Page 167 721126 LIF cartridge probe guide Table 11-2 MS Adapters Part Number Description B07363 OptiMS Adapter for SCIEX Nanospray III Source B07366 OptiMS Adapter for Thermo Nanospray II MS Source B86099 OptiMS Adapter Kit with Stage for Bruker MS B85397 OptiMS Adapter Kit with Stage for Waters MS...
  • Page 168 Deuterium lamp 144951* LIF 2-color upgrade kit A59494* Packaged laser module upgrade kit, 488 nm, single color 144094 PDA fiber-optic Y-cable B68372* Photodiode array detector upgrade 144093 UV/Vis fiber-optic cable CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 168/247...
  • Page 169 Order Parts Table 11-6 Other Parts Part Number Description A47775 Electrode A59525 Electrode tool A95348 Insertion lever interface parts kit B78399* Software reprocessing key User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 169/247...

  • Page 170: A 32 Karat Software

    The system administration feature is used to control access to the 32 Karat software and the CESI 8000 Plus software. System access can be set for a user, system, or project. When it is enabled, the system administration feature also keeps customized settings for each system.

  • Page 171: License Key

    Start the CESI 8000 Plus System Before the 32 Karat software can be used with the CESI 8000 Plus system, the system must complete an initialization process. Make sure that a cartridge is installed.
  • Page 172 32 Karat Software Figure A-1 CESI 8000 Plus Ready Window On the side menu, double-click the 32 Karat Software icon. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 172/247...

  • Page 173: Systems And Projects

    Click View > Hierarchy Pane. To show the available instruments in the right pane, click CESI 8000. Note: The Service Support option is for use by SCIEX field service employees (FSEs). A UV detector is required to use the Service Support option.

  • Page 174: Open The Direct Control Window

    The Direct Control window supplies manual control of the instrument and shows the status of instrument functions. To open the Direct Control window, click Control > Direct Control > View. Figure A-3 Direct Control Window CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 175: Create A Toolbar Icon For The Direct Control Window

    32 Karat Software Methods A method is a collection of steps that are done in a series by the CESI 8000 Plus system. A method has all of the information required to operate the instrument. Methods are created in the Instrument Setup window.

  • Page 176: Create Or Change A Method

    Use the Instrument Setup window to create a new method or run an instrument test sample in online mode. Method parameters are set on the Initial Conditions and Time Program tabs. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 177 Use the kV and μA fields to set limits for Voltage and Current. Because voltage and current are interrelated, the CESI 8000 Plus system limits both parameters when one limit is reached. For example, assume that the voltage limit is set to 30 kV and the current limit is set to 10 μA.
  • Page 178 Time Program Tab Use the Time Program tab to add the events that make up the method. Each event shows on a numbered line. The events are done in numbered order. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 179 Timed events must be in a group together. A group of timed events cannot be interrupted by an untimed event. Untimed events can only occur before a group of timed events. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module...
  • Page 180 For information about other event types, refer to the document: 32 Karat Software Help. Separate Event Use the Separate event to set the conditions for the separation. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 180/247...
  • Page 181 Set values for the selected Separation Type. Ramp Time, which is only applicable for electrical separations, sets the length of time for the voltage, current, or power to get to the value that is set. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 181/247...
  • Page 182 If the external adapter accessory is in use, then select to change the way the instrument manages the power supply. Rinse Event Use the Rinse event to clean the capillary and to load fresh buffer or other separation media. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 183 Inject Event Use the Inject event to supply sample to the capillary. Usually, the Inject event is untimed and occurs before the first Separate event. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 183/247...
  • Page 184 To change the vial positions after a specified number of cycles, select the Inlet, Outlet, or both check boxes, and then type the number of cycles to occur between changes. To see and select the tray positions graphically, click Trays. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 185: Single Run And Sequence Runs

    For single runs, a method must be manually started before every run. Single runs are useful for method development. The results of one run can suggest modifications to the method or other procedures. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 185/247...

  • Page 186: 32 Karat Software Sequences

    Note: Because a sequence table has more columns than can be shown at one time, the following Properties dialog shows the options selected for the example. Refer to the figure: Figure 7-21. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 187 Sample Inject Identifies the vial that supplies the sample for injection if the sample is in Outlet the outlet tray. In the CESI 8000 Plus system, the sample is always in the inlet sample tray. Sample ID Shows the name of the sample to be run.

  • Page 188: Sequence Validation

    Note: The instrument name must be a valid Windows folder name. For routine users to get access to an application in the CESI 8000 Plus software, the 32 Karat software project name must be the same as the 32 Karat software instrument name.
  • Page 189 Type a user name and password, and then click Save. Click OK. Click Tools > System Administration Wizard. Click User, and then click Next. Select the user name, and then click Next. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 189/247...
  • Page 190 For each of the projects to which the routine user needs access, add the Open Method, Open Sequence, Save Sequence, and Lock Instrument privileges. Note: Routine users must be given these privileges to get access to the CESI 8000 Plus software.
  • Page 191 In the Enterprise window of the 32 Karat software, right-click the instrument and then click Open. Type the user name and password, select the project, and then click Login. Click File > Sequence > Open. Click Sequence > Properties. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 191/247...
  • Page 192 Figure A-16 Method in the Sequence Run Table Click File > Sequence > Save. Do steps through for all transferred sequences. Close the instrument and the Enterprise windows. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 192/247...

  • Page 193: About The Auto-Increment Feature

    • To increment only the rinse vial at the outlet buffer tray, click Outlet. • To increment the rinse vials at the inlet and outlet buffer trays, click Inlet and Outlet. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module...
  • Page 194 Note: There are six rows in the buffer trays. Vials will not be incremented beyond row 6. Start the auto-increment with vial positions at row 1 to maximize the number of cycles. CESI 8000 Plus High Performance Separation-ESI User Guide...

  • Page 195: Set Auto-Increment For An Inject Event

    When the increment has been set, in the Increment Every field, type a value. In this example, only the inlet buffer tray will be incremented every 12 cycles, which is 12 repetitions of the same method. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 195/247...

  • Page 196: Set Auto-Increment For A Separate Event

    Start the auto-increment with vial positions at row 1 to maximize the number of cycles. Set Auto-Increment for a Separate Event In the 32 Karat software, go to the Direct Control window, and then double-click the Separate field. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...
  • Page 197 When the increment has been set, in the Increment Every field, type a value. In this example, both the inlet and outlet buffer trays will be incremented every 12 cycles, which is 12 repetitions of the same method. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 197/247...

  • Page 198: Auto-Increment In A Method

    In the following example, auto-increment is used in lines 4, 5, 7, and 8. The term In / Out vial inc 12 specifies that both the inlet and outlet vials will be incremented after 12 cycles. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 199: Review Vial Positions Before A Sequence Is Run

    Note: The CESI 8000 Plus software gives a preview of all trays before the start of a sequence. The Sequence Vials Report Preview is also used to identify vial collisions. To prevent a vial collision, change the identified vial position in the method and then save the method.
  • Page 200 Note: If the auto-increment feature is in use, then do not use different methods in a single sequence. If the sequence has different methods, then the CESI 8000 Plus system goes back to the initial set of vials specified at the start of each method.

  • Page 201: B Cesi 8000 Plus Software

    Refer to the section: License Key. If required, then the 32 Karat software can be opened from the side menu in the CESI 8000 Plus software. For information about the CESI 8000 Plus software, refer to the document: Help System.
  • Page 202 Login and Logout: Open the 32 Karat software. Note: If security is enabled for the 32 Karat software and an application and sequence or method are selected, then a login dialog opens. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 203 Description Lock and Unlock: Lock and unlock the CESI 8000 Plus software. This feature lets a user lock the controls in the CESI 8000 Plus software so that other users cannot change the controls. When the software is locked, other users can still get access to the 32 Karat software, and view, print, and export windows.

  • Page 204: Run An Application

    Note: Do not start a method or sequence run from the 32 Karat software and the CESI 8000 Plus software at the same time. The CESI 8000 Plus software will not let a user start a method or sequence run from the same connected system.
  • Page 205 When the login is successful, a connection with the system is made and the system status is updated in the top banner of the CESI 8000 Plus software. Real-time data for voltage and current shows in the Instrument Status and Direct Control pane.

  • Page 206: Samples/Vials Window

    Set Up Samples and Vials for Sequence. Set Up Samples and Vials for a Method If a method was selected in the Application window, then the method Samples/Vials window opens. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 206/247...
  • Page 207 In the Repetition field, select the number of repetitions for the method. Note: The CESI 8000 Plus software does a check for duplicate data file names. Data file names must use unique identifiers, such as the date and time or increment number.
  • Page 208 In the Samples/Vials window for a method, the Insert option can be used in the Sample ID and Data file fields to select the options to add to the file names. Figure B-6 Insert Sample ID Dialog Figure B-7 Insert Data File Dialog CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 209: Set Up Samples And Vials For A Sequence

    Click the value(s) to add to the sample ID or data file name, and then click OK. Set Up Samples and Vials for a Sequence If a sequence was selected in the Application window, then the sequence Samples/Vials window opens. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 209/247...
  • Page 210 Note: The data folder for the selected application shows as the default output data path. In the Output sequence path field, type a path, or click Browse and then select the path. CESI 8000 Plus High Performance Separation-ESI User Guide...
  • Page 211 When a sequence is run, the number of samples can be changed. Rows in the sequence table are hidden as the sample quantity decreases. When a description of the sequence is set, the User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module...
  • Page 212 B-10. Note: When the number of samples is set to 0, all rows are hidden except for rows with the Always type. Figure B-9 Sequence with All Nine Samples CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 212/247...
  • Page 213 CESI 8000 Plus Software Figure B-10 Sequence with No Samples (All Rows Hidden) User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 213/247...
  • Page 214 CESI 8000 Plus Software Figure B-11 Sequence with Five Samples CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 214/247...
  • Page 215 B-11) to four (refer to the figure: Figure B-12). To show the hidden rows that are not run, right-click the sequence, and then click Show Hidden Rows. Refer to the figures: Figure B-13 Figure B-14. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 215/247...
  • Page 216 CESI 8000 Plus Software Figure B-13 Sequence with Five Samples and Hidden Rows CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 216/247...
  • Page 217 If a range of rows is selected, then the options are applied to the selected cells. Right-click the Sample ID or Data File column, and then click Fill Down. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 217/247...
  • Page 218 Note: When the Fill Down option is selected in the Reps column, the value in the selected cell is copied into the subsequent rows. Figure B-16 Fill Down Sample ID Dialog CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 219: Display Options Menu

    Display Options. The Hide Legend, Show Tray Detail, and Show Vial Preview options are also available from the Show/Hide menu to the left of the tray view. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 219/247...
  • Page 220 The first five characters of the vial contents show in the tray view. We recommend that each of the vial content types be given a unique identifier and used consistently between methods. CESI 8000 Plus High Performance Separation-ESI User Guide Module...
  • Page 221 To change the color for the vial type, click the color next to the description and then click a new color. To change the colors in the legend to the default settings, click Reset. Hide Legend Hides the vial legend. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 221/247...
  • Page 222 Shows the full tray detail set by the tray view (Program Event or Vial Detail Contents). The vials can be shown as circles or rectangles. The tray detail view can be exported and printed. Figure B-19 Tray Detail View CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 222/247...

  • Page 223: Acquisition Window

    • If a sequence is active, then the Sequence Run table shows which row of the sequence is active. • The Current Run table shows which event in the method is active. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 223/247...

  • Page 224: During A Run

    During a Run During a run, the Running Acquisition window is empty. The system state is Scheduled Run. The countdown until the start of the scheduled run shows below the system state. CESI 8000 Plus High Performance Separation-ESI User Guide Module...

  • Page 225: Stop A Run

    To see the settings in the Application and Samples/Vials windows, click 1. Application, 2. Samples/Vials, Back, or Next. After a run has started, the settings cannot be changed. Only one sequence or method at a time can be started from the CESI 8000 Plus software. Stop a Run Note: A method or sequence that has been stopped cannot be started again.

  • Page 226: Complete A Run

    ID as text on top. The All Runs tab shows the sample ID in the key for the trace. Multiple Colors When selected, opens each trace in a different color. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 227: Describe Sequence Window

    For a routine user to change the number of samples used in a sequence, the sequence must be described in the CESI 8000 Plus software. Note: For a routine user to describe a sequence, the user must have the Sequence File Write privilege in the 32 Karat software.
  • Page 228 Figure B-24 Describe Sequence Window Without Types If a sequence is changed in the 32 Karat software after being described in the CESI 8000 Plus software, then the changes are detected and only the full sequence can be run. To change the sample quantity, the sequence must be described again in the CESI 8000 Plus software.

  • Page 229: Describe The Sequence

    CESI 8000 Plus Software Describe the Sequence In the CESI 8000 Plus software main window, click Figure B-25 Describe Sequence Window In the Application field, click the application. In the Sequence field, click the sequence to describe. If a prompt is shown, then type the user name and password.
  • Page 230 Note: If a user tries to change the text in the brackets, then the described sequence will be invalidated. The sequence must be described and saved again. Figure B-26 32 Karat Software Sequence Editor Click Finish. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E...

  • Page 231: C Connect An Ethernet Cable

    Connect an Ethernet Cable Connect an Ethernet Cable An Ethernet cable can be used to connect the CESI 8000 Plus system and the mass spectrometer. Turn on power to the CESI 8000 Plus system and controller. Connect one end of the Ethernet cable to the CESI 8000 Plus system controller and the other end to the acquisition computer on the mass spectrometer.
  • Page 232 Connect an Ethernet Cable Note: The RS-232 Service Only connector on the CESI 8000 Plus system is to be used by a SCIEX field service employee (FSE) only. Figure C-2 RS-232 Service Only Connector CESI 8000 Plus High Performance Separation-ESI...

  • Page 233: D System Specifications

    Temperature: 15 °C to 40 °C (59 °F to 104 °F), 15 °C to 30 °C (59 °F to 86 °F) recommended Maximum heat dissipation 400 W (1,024 BTU/hr) Pollution degree TTL: 2 Contact closures: 2 User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 233/247...

  • Page 234: Supplied Controller Configuration

    System Specifications Supplied Controller Configuration Note: Specifications are subject to change without notice. Note: SCIEX fully validates and supports the controllers supplied with the system. Only limited support is available for customer-supplied computers. Table D-2 Supplied Controller Configuration Item Details...

  • Page 235: Capillary Temperature Control Specifications

    Injection: 0.1 psi to 25 psi (pressure) or 0.1 psi to 5.0 psi (vacuum) Rinse: 0.1 psi to 100 psi (pressure) or 0.1 psi to 5.0 psi (vacuum) Stability ±0.3 psi at 25 psi ±1.0 psi at 100 psi User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 235/247...

  • Page 236: Detector Specifications

    Filter dimensions Diameter: 12.7 mm (0.5 inches) Thickness: 5 mm (0.2 inches) UV source 30 W pre-aligned deuterium lamp Wavelength accuracy ±2 nm Wavelength range 190 nm to 600 nm CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 236/247...

  • Page 237: (Optional) Lif Detector Specifications

    Laser output delivered to the capillary: 2.5 mW ±0.5 mW Laser wavelength: 488 nm nominal Wavelength range (for Excitation: 300 nm to 700 nm optics) Detection: 350 nm to 750 nm User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 237/247...

  • Page 238: (Optional) Pda Detector Specifications

    Scan collection frequency 0.5 Hz to 32 Hz UV source 30 W pre-aligned deuterium lamp UV source lifetime 1,000 hours Wavelength accuracy 2 nm Wavelength range 190 nm to 600 nm CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 238/247...

  • Page 239: E Glossary Of Symbols

    Alternating current Amperes (current) Asphyxiation Hazard Authorized representative in the European community Biohazard CE Marking of Conformity cCSAus mark. Indicates electrical safety certification for Canada and USA. Catalog number User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 239/247...
  • Page 240 Symbol Description Caution. Consult the instructions for information about a possible hazard. Note: In SCIEX documentation, this symbol identifies a personal injury hazard. China RoHS Caution Label. The electronic information product contains certain toxic or hazardous substances. The center number is the Environmentally Friendly Use Period (EFUP) date, and indicates the number of calendar years the product can be in operation.
  • Page 241 Hertz International safety symbol "Caution, risk of electric shock" (ISO 3864), also known as High Voltage symbol If the main cover must be removed, then contact a SCIEX representative to prevent electric shock. Hot Surface Hazard In Vitro Diagnostic Device...
  • Page 242 Relative humidity must not exceed 99%. Keep upright. Lacerate/Sever Hazard Laser Radiation Hazard Lifting Hazard Magnetic Hazard Manufacturer Moving Parts Hazard Pacemaker Hazard. No access to people with pacemakers. CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 242/247...
  • Page 243 Transport and store the system within the specified minimum (min) and maximum (max) levels of relative humidity, noncondensing. Transport and store the system within –30 °C to +45 °C. User Guide CESI 8000 Plus High Performance Separation-ESI RUO-IDV-05-3897-E Module 243/247...
  • Page 244 UKRP United Kingdom Responsible Person Volt Ampere (apparent power) Volts (voltage) WEEE. Do not dispose of equipment as unsorted municipal waste. Environmental Hazard Watts (power) yyyy-mm-dd Date of manufacture CESI 8000 Plus High Performance Separation-ESI User Guide Module RUO-IDV-05-3897-E 244/247...

  • Page 245: F Glossary Of Warnings

    Glossary of Warnings Glossary of Warnings Note: If any of the labels used to identify a component become detached, then contact a SCIEX field service employee (FSE). Label Translation (if applicable) FOR RESEARCH USE ONLY. NOT FOR USE FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.

  • Page 246: Contact Us

    SCIEX Now Learning Hub Purchase Supplies and Reagents Reorder SCIEX supplies and reagents online at store.sciex.com. To set up an order, use the account number, found on the quote, order confirmation, or shipping documents. Currently, customers in the United States, Canada, United Kingdom, Belgium, Netherlands, France, Germany, and Switzerland have access to the online store, but access will be extended to other countries in the future.
  • Page 247 Contact Us To find hardware product documentation, refer to the documentation that comes with the system or component. The latest versions of the documentation are available on the SCIEX website, at sciex.com/ customer-documents. Note: To request a free, printed version of this document, contact sciex.com/contact-us.

Table of Contents