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Waters Prep 80q SFC System Manual

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Prep 80q SFC
System Guide for ChromScope v1.2.1
715004623 / Revision A
Copyright © Waters Corporation 2014
All rights reserved

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Summary of Contents for Waters Prep 80q SFC

  • Page 1 Prep 80q SFC System Guide for ChromScope v1.2.1 715004623 / Revision A Copyright © Waters Corporation 2014 All rights reserved...

  • Page 2: General Information

    General information Copyright notice © 2014 WATERS CORPORATION. PRINTED IN THE UNITED STATES OF AMERICA AND IN IRELAND. ALL RIGHTS RESERVED. THIS DOCUMENT OR PARTS THEREOF MAY NOT BE REPRODUCED IN ANY FORM WITHOUT THE WRITTEN PERMISSION OF THE PUBLISHER.

  • Page 3: Contacting Waters

    Contacting Waters Contact Waters with enhancement requests or technical questions regarding the use, transportation, removal, or disposal of any Waters product. You can reach us via the Internet, telephone, or conventional mail. Waters contact information Contacting Information medium Internet The Waters Web site includes contact information for Waters locations worldwide.

  • Page 4: High Voltage Hazard

    High voltage hazard Warning: To avoid electric shock, do not remove protective panels from system modules. The components within are not user-serviceable. Warning: To avoid nonlethal electric shock when the instrument is in Operate mode, avoid contact with the areas marked with the high-voltage warning symbol.

  • Page 5: Applicable Symbols

    Audience and purpose This online information system is for personnel who install, operate, and maintain the Prep 80q SFC system. It provides most of the information required to set up, operate, and maintain the system's hardware and software components. The only...

  • Page 6: Intended Use For A Prep Sfc System

    Waters 2535/2545/2555 Quaternary Gradient Module Operator's Guide. Intended use for a Prep SFC system Waters designed the Prep SFC system to serve as a high-pressure separator for quantitative recovery of purified products such as enantiomers, complex synthetic chemicals, and natural products, or for research. It is not intended for use in diagnostic or biologically hazardous applications.

  • Page 7: Table Of Contents

    Audience and purpose ..............v Intended use for a Prep SFC system..........vi ISM classification: ISM group 1 class A ..........vi Getting started with the Prep 80q SFC system ......15 Supercritical fluid chromatography.............15 1.1.1 Prep 80q SFC system flow ............. 15 Prep 80q SFC system components .............17...
  • Page 8 2.1.4 Lifting ..................28 Gas supply (liquid CO ) requirements............29 Solvent requirements................29 Laboratory ac power requirements.............30 2.4.1 System and component power requirements ........30 2.4.2 Plug types ................... 31 General environmental requirements............32 Temperature requirements ...............32 Vibration considerations ................32 Magnetic field considerations..............32 Radio emission considerations ..............32 2.10 Safety considerations ................33...
  • Page 9 3.4.2 Removing a project............... 44 Administration ................45 User management ...................45 4.1.1 Specifying security preferences ............45 4.1.2 Creating a user account..............46 4.1.3 Configuring a user account ............46 4.1.4 Removing a user account .............. 46 Specifying the security mode for the ChromScope workstation.......46 Method and sequence development...........
  • Page 10 System startup ..................83 6.2.1 Starting the system hardware ............84 6.2.2 Starting the ChromScope software and Waters Instrument Console..84 6.2.3 Confirming the integrity of column-tubing connections and the MV2 valve position ................85 6.2.4 Confirming that CO and co-solvent supplies are ready to use ... 86 6.2.5...
  • Page 11 8.2.2 Specifying peak-information options..........118 8.2.3 Specifying options for displaying run information ......120 8.2.4 Specifying graph display options........... 122 8.2.5 Setting annotation display options and orientation ......124 8.2.6 Specifying overlay options and axis offsets in overlay ...... 124 8.2.7 Setting display preferences for PDA options ........
  • Page 12 Submitting data-log files for batch processing ........... 154 Configuring auto report mail ..............157 10 System maintenance ............... 158 10.1 Maintenance schedule ................158 10.2 System shutdown preparation ..............160 10.2.1 Shutting down the system for brief periods ........160 10.2.2 Shutting down the system for long periods ........
  • Page 13 Safety advisories ..............187 Warning symbols................... 187 A.1.1 Specific warnings................ 188 Notices ....................190 Warnings that apply to all Waters instruments and devices ......190 Electrical symbols .................. 194 Handling symbols .................. 195 Specifications ................196 Prep 80q SFC system specifications............196 High pressure CO and co-solvent pumps specifications ......
  • Page 14 Collection-bottle cabinet specifications............198 July 28, 2014, 715004623 Rev. A Page xiv...

  • Page 15: Getting Started With The Prep 80Q Sfc System

    SFC is suitable for separating and isolating chiral and achiral chemical compounds. Bulk-scale purification systems, such as the Waters Prep 80q/200q/200BTq/350q SFC systems, are designed to serve as high-pressure separators for quantitative recovery of purified products such as enantiomers, complex synthetic chemicals, and natural products.
  • Page 16 Figure 1–1: Prep 80q SFC system flow diagram Sample prep injector Co-solvents Mixing tee P-50 Flow meter Dampener Prep column Cooling UV detector Liquid CO ABPR Seven low-pressure column separators to collect fractions exhaust and waste Fractions Waste P-50 pump, standard flow rate of 50 g/min...

  • Page 17: Prep 80Q Sfc System Components

    The Prep80q SFC system components are rack-mounted for placement on a laboratory benchtop, as shown in the representation of the base system below. Figure 1–2: Prep 80q SFC system components The Prep 80q SFC system, under ChromScope control, comprises these major components: July 28, 2014, 715004623 Rev. A...
  • Page 18 Represents the sample-injection module, which injects one sample at a time using a standard 2-mL loop and 5-mL syringe. The module is preset at the factory for modifier-stream injection. Nevertheless, a Waters service representative can switch the module to support combined-stream injection.
  • Page 19 Item Description Represents the P-50 high-pressure pump module, which pumps the liquefied carbon dioxide to the Prep 80q SFC system. The module's key features are these: • Dual pump heads with cooling chambers for removing the heat of compression, to keep the CO...
  • Page 20 Component Description Prep column The standard column supplied with the Prep 80q SFC system, a Viridis Silica column, 5 µm, 50 × 150 mm, is used to validate the system during installation and maintenance. July 28, 2014, 715004623 Rev. A...

  • Page 21: Safety Practices And Warnings

    Component Description Chiller unit and The Thermo Scientific Accel 500 Laboratory Chiller provides a cooling bath continuous supply of fluid at a constant temperature and components volume. The unit consists of an air-cooled refrigeration system, cooling heat exchanger (HE1), recirculating pump, polyethylene reservoir, and a microprocessor controller.

  • Page 22: Typical System Workflow

    To avoid creating a hazardous condition in which a collection bottle can explode, should it become pressurized, avoid these conditions: • Use of alternate-sized collection bottles without Waters' approval • Use of bottles that are not safety-coated • For the exhaust tubing, any size reductions, sharp bends, kinks, or other...

  • Page 23: Selecting And Running A Method-Screening Sequence

    1.4.1 Selecting and running a method-screening sequence To fine tune the purification chromatography on the system and store the optimal settings in a data acquisition file, you select and start a method-screening sequence, or scouting run. Afterwards, you can use the file's plots to easily calculate and specify the fraction-collection and injection-timing settings used to build the stacked- injection purification sequence.

  • Page 24: Starting And Monitoring A Run

    Tip: Typically, when specifying the method and sequence settings for a purification run, you quantitate the amounts of purified compound and co- solvent that you expect to recover during the run, to avoid overfilling the fraction collection vessels and losing valuable product. In the collection method for the purification run, specify the settings for directing how you will collect the fractions (detector signal threshold, slope threshold, or timed collection) and which collection vessels you will use to...

  • Page 25: Analyzing The Run's Data For Quality Control

    Prepare the system for the solvents, sample, and collection vessels required by the purification run. Load and start the purification sequence. From the ChromScope Main window and System Panel, monitor the run, as needed. Tips: • To extend the current run so that you can maintain system equilibration while performing maintenance tasks such as refilling solvents or emptying collection vessels, click Pause .
  • Page 26 Tip: On the basis of this information, you can evaluate, for purposes of quality control, the effectiveness of settings, instrumentation, and processes, and you can determine what changes, if any, are needed to achieve best results. July 28, 2014, 715004623 Rev. A Page 26...

  • Page 27: Component Connections

    Component connections 2.1 Installation and operation requirements You must plan well before moving the system to a different area, replacing its modules, or modifying your laboratory facility. 2.1.1 System and component dimensions and weights Minimum positioning and space requirements for the system appear in the following table.

  • Page 28: Additional Space Considerations

    System Width Depth Height Weight component Accel 500 chiller 22.7 cm 48.7 cm 60.3 cm 29 kg unit Requirement: The chiller unit, located on the floor next to the system, requires a minimum of 30.5 cm of clearance at the front and rear, for ventilation.

  • Page 29: Gas Supply (Liquid Co 2 ) Requirements

    • Assess the risk of injury. • Act to eliminate risk. • Plan the operation in advance and in conjunction with a Waters engineer. • Follow appropriate country or company regulations. • Do not, under any circumstance, jar, shake, strike, or subject to collision or vibration any instrument or device.

  • Page 30: Laboratory Ac Power Requirements

    • Waters recommends the use of a line conditioner or an uninterruptible ac power supply (UPS), for optimum long-term input-voltage stability. 2.4.1 System and component power requirements...

  • Page 31: Plug Types

    System Input voltage/ Fuse rating Rated power component frequency draw ABPR 100-240 Vac, 50/60 3A - 250V 220 VA 2489 UV Visible 100-240 Vac, 50/60 3.15 A 185 VA detector Six-zone 100-240 Vac, 50/60 1A - 250V 22 VA temperature controller module (CN6) Heat exchanger...

  • Page 32: General Environmental Requirements

    2.5 General environmental requirements Refer to these general environmental requirements when preparing the laboratory: • Waters recommends you install the system in an environmentally controlled laboratory in a draft-free location away from excessive amounts of dust. • Do not locate the system directly beneath air-conditioning or heating ventilation.

  • Page 33: Safety Considerations

    2.12 Plumbing connections A Waters technician equips and plumbs a system as part of its setup. All tubing connections between components are prefitted and labeled for their intended use.

  • Page 34: Fittings

    Waters service representative for assistance. Many fittings appear identical or similar. Use only the specified ferrule and gland nut for the fitting. If you are unsure of a particular type of fitting, contact your Waters service representative for assistance. 2.12.3 External CO...

  • Page 35: Co Pump Cooling Tubing

    Dip tube liquid 2.12.4 CO pump cooling tubing The CO pump requires external cooling for proper operation. The connection between the outlet of the cooling tubes that serve the pump heads and the inlet to the cooling heat exchanger is preconfigured. The cooling heat exchanger is plumbed to the chiller unit, as part of the initial system setup.

  • Page 36: Co Exhaust Connections

    2.12.5 CO exhaust connections You must plumb the flexible ducting from the low-pressure collection cabinet to a 10.2-cm vent port, for proper venting of the gaseous CO and solvent waste residue. A 20-L HDPE bottle and interconnecting tubing, provided with the system, contains liquid waste.

  • Page 37: Electrical Considerations

    If you move the system to an area where the line voltage differs from that in the original location, you must reconfigure the system for the new voltage. In such a case, contact your Waters Service Representative for information about changing the operating voltage.

  • Page 38: Inter-Module Signal Connections

    All necessary cables are provided along with the system, labeled for their intended use, and connected by a Waters service representative during the initial system setup. These connections are mapped to specific COM ports in the PC workstation, which are listed in the following table.

  • Page 39: Getting Started With Chromscope Software

    Requirement: When the software initially opens, you must provide these default settings: • Login: Administrator • Password: waters 3.2 Exiting from ChromScope software To exit from the ChromScope software: From the File menu, click Exit. July 28, 2014, 715004623 Rev. A...

  • Page 40: Chromscope Software Interface

    3.3 ChromScope software interface The ChromScope main window contains a view of the projects, system status, and configuration components, a system control panel, and the menus and controls necessary to manipulate those software elements and navigate the application. Nodes in folder view provide controls and options for the files within it Menu bar Toolbar System Status Panel shows each module's status at a glance...
  • Page 41 Sequence, instrument method, and collection-method panels for making on-the- fly (OTF) updates (when operating the ChromScope software in non-compliant mode) Real-time display of acquisition System message box lists system status and individual module status when you move the pointer over it File list view area displays list of files in project's subfolder node The ChromScope software provides simple work flows for developing methods and collecting fractions.

  • Page 42: Project Management

    Table 3–1: ChromScope key components and functions (continued) Component or Purpose function Method and Specify and save the parameter settings of system modules, sequence editors fraction collection components, run duration, and for governing chromatogram integration. Later, you can edit or modify the methods and sequences, as needed, load sequences into a queue, and run them to perform acquisitions or purify sample.

  • Page 43: Setting Up A New Project

    The table "ChromScope subfolder structure" describes all of the project's subfolders and the files they contain. Table 3–2: ChromScope subfolder structure Subfolder Description Sequence Files Contains sequences or the files that contain the options for injecting a sample or series of samples. Method Files Contains sub-nodes for Instrument Methods, Integration Methods, and Collection Methods.

  • Page 44: Removing A Project

    Rule: After you create and save your new project, you cannot change its subfolder-naming convention. The following table lists and describes the subfolder conventions that you can choose from. Table 3–3: Subfolder naming conventions Subfolder File organization naming convention Date Places all data-log files created in the project in a subfolder labeled with the date on which each file was created.

  • Page 45: Administration

    Administration 4.1 User management As a ChromScope system administrator, you can use the Account Management function to accomplish these tasks: • Control access to the ChromScope software functions by establishing and maintaining user accounts. • Enforce limits on what ChromScope components a user can see and do by assigning the user specific component rights.

  • Page 46: Creating A User Account

    When you clear the check box, the log-in screen is hidden, and audit trail functionality for generated logs or reports is disabled. 4.1.2 Creating a user account To create a user account: From the Tools menu, click Account Management > User Manager. In the User List area of the User Manager window, right-click, and then click Add User.
  • Page 47 compliant mode, the OTF functionality is unavailable, and the current method and sequence settings are read-only. In the non-compliant security mode, workstation users can also display, or hide, the OTF update panels, on the right-hand side of the System Panel Additionally, for convenience, they can specify preferences for choosing which update options are applied by default.

  • Page 48: Method And Sequence Development

    Method and sequence development 5.1 Methods for controlling system functions In the instrument method, you specify and save the settings that control the rate and composition of the mobile phase, system pressure, column temperature, detector wavelength, and other important run conditions. Later, you can edit the method, as needed, add it to a sequence, and use it to run acquisitions or purification runs.

  • Page 49: Viewing Or Editing An Instrument Method

    pressure. This capability is especially useful when cleaning a column or establishing a baseline. • Use the instrument method settings in concert with those specified under the System Configuration node, to protect system components from unsafe conditions, such as excessive pressure drop or column overtemperature. The Instrument Method editor in the ChromScope software has tabs on which you configure the individual components that comprise a system.
  • Page 50 On the General Method tab, enter a name for the method and, optionally, a brief description of the method's purpose. Enter the remaining general method settings referring to the table "General method settings". Important: You can access the Save button only from the General Method tab.
  • Page 51 Composition Programs the composition of the mobile phase by specifying the percentage of co-solvent for the Total Flow. For isocratic separations, the default condition, the Co-Solvent % setting defines the percentage of co-solvent for the duration of the method. For a composition gradient, it is the Co-Solvent Start % setting for the first stage of the gradient.
  • Page 52 Back Sets the system operating pressure, in bars, using the ABPR. pressure For isobaric separations, the default condition, the Total Back Pressure setting defines the operating back pressure for the duration of the method. Less common in SFC is the use of a pressure gradient.
  • Page 53 With the instrument method open, click the W2545QGM Pump tab. In the W2545QGM Pump tab, enter the settings for the co-solvent pump referring to the following table. Tip: For further details about configuring the 2545 QGM, refer to the Waters 2535/2545/2555 Quaternary Gradient Module Operator's Guide. Setting or Purpose...
  • Page 54 5.1.2.3 Configuring injection module parameters To configure the injection module parameters: With the instrument method open, click the Injector tab. On the Injector tab, specify values for the injection module referring to the table "Injector method settings". Table 5–2: Injector method settings Setting or Purpose control...
  • Page 55 Table 5–2: Injector method settings (continued) Setting or Purpose control Sample Save Specifies the volume for the connector tubing, which is Volume typically 0.1 mL. Tip: To avoid sample loss, if you change the factory- configured connector tubing to one of a different size, change the Sample Save Volume setting accordingly.

  • Page 56: Exporting An Instrument Method

    Table 5–3: 2489 UV/Visible detector method settings (continued) Setting or Purpose control Auto Zero on Optionally used to set the absorbance offset so that the Inject Start analog output reads 0 AU each time the detector receives an inject-start signal via a contact closure, Ethernet, or through the detector's front panel.

  • Page 57: Viewing Or Editing An Integration Method

    from the list, and choose whether to apply the method to all chromatograms in the file or just the currently displayed chromatogram. 5.2.1 Viewing or editing an integration method You view or edit an integration method using the Integration Method Editor. To view or edit an integration method: Click the Integration Method node, to display the currently stored integration methods.
  • Page 58 Table 5–4: Integration method settings (continued) Setting or Purpose control Time Window Specifies a portion of the run's duration for processing the Start and peak data acquired from the injection. The default values of Time Window "0" for the Time Window Start and Time Window Stop Stop functions allow the entire acquisition file, from inject start to the end of the run, to be processed.
  • Page 59 Table 5–4: Integration method settings (continued) Setting or Purpose control Threshold For threshold integration only, specifies the absorbance Start and settings used to start and stop integration of the peak. Threshold Tip: Use with Time Window Start and Time Window Stop Stop parameters to define a window when the software will start looking for the signal strength to start and stop integration.
  • Page 60 Table 5–4: Integration method settings (continued) Setting or Purpose control Peak Name Assigns names to peaks, for easier peak identification in a chromatogram and for use when determining concentration by peak name. Enter the peak names and their corresponding retention-time settings and offset settings in the Peak Name data grid.

  • Page 61: Exporting An Integration Method

    Table 5–4: Integration method settings (continued) Setting or Purpose control Smoothing For second-derivative integration only, improves the degree presentation and interpretation of a chromatogram by improving the signal-to-noise ratio, to prevent the software from identifying baseline noise as peaks. You can specify settings of none, 5, 7, and 9 through 31.

  • Page 62: Sequences For Acquiring Data And Collecting Fractions

    To export an integration method: Click the Integration node, to display the currently stored integration methods. Right-click the file that you want to export, and select Export To and the destination project from the list. Result: The integration method appears in the new project location. 5.3 Sequences for acquiring data and collecting fractions In a sequence, you specify and save the methods, run times, injection volumes, and...

  • Page 63: Viewing Or Editing A Sequence

    • You can define any number of rows in the table. • Required fields are denoted with an asterisk. When following the instructions in this topic, always observe Good Laboratory Practice (GLP) guidelines and your laboratory's standard operating procedures. 5.3.1 Viewing or editing a sequence You view or edit a sequence using the Sequence Editor.
  • Page 64 Table 5–5: Sequence settings Setting or Purpose control Injection # Identifies the number and order of the injection, which becomes a step in the running sequence. Run Dur. Specifies the duration of the run, in minutes, after injection. Inj. Delay Specifies a wait time, in minutes, before injecting the sample.

  • Page 65: Creating A Method-Screening Sequence

    Table 5–5: Sequence settings (continued) Setting or Purpose control Integration Used during a running sequence to perform a batch run-and- Method and report queue. After the execution of each step in the running Report sequence, you can automatically generate and print a report Template on the PC's default printer based on the integration and report formats defined in the Integration Method and Report...
  • Page 66 Rule: This setting is automatically selected when you build a method- screening sequence or stacked injection sequence using the Sequence wizard. Click Sequence Wizard > Method Screening, and then click Next. Enter the remaining sequence settings referring to the table at the end of this procedure.

  • Page 67: Building Purification Sequences

    Table 5–6: Method-screening sequence settings (continued) Setting or Purpose control Integration Used during a running sequence to perform a batch "run Method, Report and report" queue. Template, and After the execution of each step in the running sequence, or Print Options after the entire sequence, you can automatically generate and print a report on the PC's default printer according to the integration and report formats defined in the...
  • Page 68 Requirement: To create a stacked-injection sequence you need this information: • A reproducible chromatogram with known peaks and retention times, for data collection. • An understanding of how these parameters are used for the sample collection chromatogram: • Stacked cycle time, or the time from the front of the desired peak set elution to the end of the peak set.

  • Page 69: Building A Purification Sequence Using The Wizard

    5.3.5 Building a purification sequence using the wizard To build a purification sequence using the wizard: In the Folder/File view, for the project, navigate to the Sequence Files node. Right-click the Sequence Files node, and click Add new sequence and then Yes.
  • Page 70 Setting or Purpose control Instrument Specifies the instrument method used to control the Method system's run conditions for the injection. Click the list to view all of the instrument methods defined for the project. As you move the pointer over each method in the list, a brief description of the method appears.

  • Page 71: Exporting A Sequence

    Setting or Purpose control Integration Used during a running sequence to perform a batch "run and Method, Report report" queue. Template, and After the execution of each step in the running sequence, or Print Options the entire sequence, you can automatically generate and print a report on the PC's default printer according to the integration and report formats defined in the Integration Method and Report Template fields of the sequence.

  • Page 72: System Configuration And Startup

    System configuration and startup 6.1 System and module settings In the System Configuration panels, you define your preferences for shutting down the system in response to certain operating conditions and for configuring the individual module's alert thresholds and usage settings. You can also view a module's real-time device status for maintenance or diagnostic purposes.
  • Page 73 Table 6–1: System shutdown options (continued) Setting or Purpose control Stop system after Maintains the current run conditions (flows, temperature, x min. of inactivity and other conditions) after the last run ends or if you clear the sequence queue. Enter the desired time delay before the software halts the system.

  • Page 74: Configuring Default Update Settings

    Table 6–1: System shutdown options (continued) Setting or Purpose control Auto wash Injector Automatically triggers the injection module to wash the injection syringe and sample loop after the last run ends (or if you clear the sequence queue). Important: The injection wash returns through the sample line to the sample reservoir.
  • Page 75 To configure the default update settings: Click the Default Update Settings tab, to open the Update panel. In the Update panel, click the option that you want for each of the update categories. (See the table at the end of this procedure.) Note: These are the default options that are automatically selected for the appropriate category when you click Apply in the open current method or...
  • Page 76 Table 6–2: OTF update options (continued) Update Option Purpose category Sequence Apply changes For adding or deleting rows in the current to current sequence. sequence Tip: This setting is typically used for extending a running sequence or changing its injection volumes. Apply changes For adding or deleting rows in the current to all matching...

  • Page 77: Configuring Settings Or Viewing Device Status For The Co 2 Pump

    Attenuation Factor Typically set to 1. (For use by Waters service technicians only.) Proportional Constant Typically set to 3. (For use by Waters service technicians only.) Integral Constant Typically set to 4. (For use by Waters service technicians only.) Integral Max Typically set to 8500.

  • Page 78: Configuring Control Settings For The Injector

    Table 6–4: CO pump PID settings (continued) Setting or Control Purpose Derivative Constant Typically set to 2. (For use by Waters service technicians only.) 6.1.4 Configuring control settings for the injector To configure control settings for the injector: Click the Injector tab.

  • Page 79: Configuring Settings Or Viewing Device Status For The Heater Controller

    Cycle Time Typically set to 5. (For use by Waters service technicians only.) Dead Band Typically set to 0. (For use by Waters service technicians only.)
  • Page 80 Table 6–6: Heater controller settings - Zone 1 (continued) Setting or Purpose control Derivative Typically set to 1. (For use by Waters service technicians only.) Upper Integral B Typically set to 63. (For use by Waters service technicians only.) Lower Integral B Typically set to 63.

  • Page 81: Manually Igniting And Extinguishing The Detector Lamp

    Cycle Time Typically set to 5. (For use by Waters service technicians only.) Dead Band Typically set to 0. (For use by Waters service technicians only.)

  • Page 82: Configuring Settings Or Viewing Device Status For The Abpr

    Rule: When this alarm threshold is exceeded, ABPR operation stops. High pressure alarm Normally set to 350 bar. (For use by Waters service technicians only.) Rule: When this alarm threshold is exceeded, ABPR operation stops. July 28, 2014, 715004623 Rev. A...

  • Page 83: Configuring Settings For The Collection Controller

    Table 6–10: ABPR pressure regulator threshold and usage settings (continued) Setting or control Purpose Attenuation Factor Typically set to 1. (For use by Waters service technicians only.) Proportional Typically set to 300. (For use by Waters service Constant technicians only.) Integral Constant Typically set to 500.

  • Page 84: Starting The System Hardware

    After the system hardware is powered-on, you can log in to ChromScope software, to monitor components and set initial conditions. For additional device status for the 2489 UV/visible detector and 2545 QGM, you can launch the Waters Instrument Console for each of those modules.

  • Page 85: Confirming The Integrity Of Column-Tubing Connections And The Mv2 Valve Position

    6.2.3 Confirming the integrity of column-tubing connections and the MV2 valve position Each time you start the system, inspect the column-tubing fittings and MV2 valve position, to ensure the system is ready for operation. To confirm the integrity of column-tubing connections and MV2 valve position: Inspect the column tubing connections, to ensure that the column or unions are securely connected, leaving no open flows.

  • Page 86: Confirming That Co And Co-Solvent Supplies Are Ready To Use

    Quaternary Gradient Module Instrument Console software, and follow the instructions for priming the QGM and seal-wash pump in the Waters 2535/2545/2555 Quaternary Gradient Module Operator's Guide. 6.2.5 Warming the detector's lamp Before performing an injection, allow the detector's lamp to warm for a minimum of 30 minutes, to stabilize its optics.

  • Page 87: Priming The Injection Module

    If you must use vessels of a different size, before you install them, ensure that Waters approves the vessels for use, because some vessels can require safety modifications.

  • Page 88: Specifying The Initial Conditions For A Purification Run And Verifying Operational Readiness

    To confirm the collection vessels, exhaust lines, and waste container are ready for operation: Inspect the collection vessels for cracks, chips, or abrasions to the safety coating, replacing any of those found defective. Inspect the exhaust lines for kinks or obstructions. Requirement: To ensure the CO can exhaust properly, rinse and clean all...

  • Page 89: System Status

    To quickly verify whether the system can achieve and maintain a pressure set point of 100 bar: Start a method in which the total flow rate is 40 mL/min, and the ABPR set point is 100 bar. View the pressure indicator and plot in the ChromScope System Panel and Current Run tab.

  • Page 90: Module Status

    • In the System Panel, when you move the pointer over each of the module icons, the System Messages box displays additional, module-specific status and parameter details. • To scroll upwards or downwards through the status and system messages in the System Messages box, use the mouse's wheel.
  • Page 91 Table 6–11: P-50 high-pressure pump module status (continued) Status Indicator location Set Flow The Set Flow indicator in the System Panel's System Messages box for the Pump A CO module shows, in g/min, the specified flow-rate set point for CO Note: To calculate the set-flow value, in the active instrument method.
  • Page 92 Quaternary Gradient Module Instrument Console software. • For additional details about the module's operating state and LCD status panel, refer to the Waters 2535/2545/2555 Quaternary Gradient Module Operator's Guide. Set Flow The Set Flow indicator in the System Panel's System Messages box for the Pump B module shows the set point for the flow rate, in mL/min.
  • Page 93 Table 6–12: 2545 QGM status (continued) Status Indicator location A, B, C, The LCD display on the module's front panel, and the A, B, C, and D and D indicators in the System Panel's System Messages box for the Pump B module, show the percentage of solvents from as many as four solvent reservoirs that can make up the composition of the co-solvent.
  • Page 94 Status Indicator location Operating The Enabled LEDs, Z1 through Z6, on the module's front panel glow state steady green when the represented zone is actively controlling its designated heat exchanger module. These are the Enabled LED's other operational modes: • Flashing green during operation indicates the module is working to maintain the temperature set point, alternately activating and deactivating the heat exchanger, as needed.
  • Page 95 6.3.1.5 Determining the status of the 2489 UV/visible detector Table 6–13: Detector status indicators Status Indicator location Power/ When the detector is powered-on and working properly, its display operating panel is lit and displays the module's operating state and program state functions.
  • Page 96 Status Indicator location Operating The Enabled LED appears on the left-hand side of the module's state front panel, above the Power LED. When the module is powered-on and working properly, the Enabled LED glows steady green. These are the Enabled LED's other operational modes: •...
  • Page 97 Status Indicator location Needle The Needle Position indicator in the System Panel's System Position Messages box for the Back Pressure module shows the extent of the valve's open position. (6000 is fully opened; 0 is fully closed.) Important: When the ABPR is idle, from the System Configuration node's Pressure Regulator tab, you can use the "Set Needle Count"...

  • Page 98: Data Acquisition And Fraction Collection

    Data acquisition and fraction collection 7.1 Loading sequences Sequences and the instrument and collection methods that they reference control the conditions and sequence of steps that you follow when performing acquisitions and sample purifications. They also control system functions for maintenance or troubleshooting procedures.

  • Page 99: Starting And Stopping A Run

    To load sequences into the sequence queue: In the Folder/File view, under your project folder, click the Sequence Files node to display the sequence files that you want to load in preparation for running the system. For each sequence you want to load, right-click the sequence, and click Open. In the sequence editor, click Add to Queue.
  • Page 100 Table 7–1: System Control panel (continued) Control Function Temporarily halt a running sequence or instrument method. Pause Result: The current sequence's run time counter stops, and any running gradient is paused. Yet the current settings for temperature, pressure, and CO and co-solvent flows remain unaltered.
  • Page 101 Table 7–1: System Control panel (continued) Control Function Restart the Click to restart the current step in the sequence. Current Step Skip to the Click to skip to the next sequence in the queue. Next Sequence Tips: • During a method-screening run, when you find the optimum conditions and want to quit the sequence, if no other sequences are in the queue, click Skip to the Next Sequence instead of Pause...

  • Page 102: On-The-Fly Updates

    Table 7–2: System operation statuses (continued) Indicator Systems's operating state Stopped 7.3 On-the-fly updates Using the spontaneous, or on-the-fly (OTF), update function, you can make changes to runs in a sequence or queue by opening the current method or sequence; updating the settings;...
  • Page 103 Figure 7–1: Displaying or hiding the OTF update panels Figure 7–2: Example Default Update Settings When you apply updates to a run, sequence, or method, you can accept the default setting selected on the Default Update Settings panel or, if the Show these options on update check box is selected, select a different option before you click Apply.

  • Page 104: Updating A Run's Settings Directly From The Otf Update Panels

    7.3.1 Updating a run's settings directly from the OTF update panels From the settings on the OTF update panels, for the current injection of a non- stacked sequence, you can directly change the sequence or method settings for the run. Restriction: You cannot save updates to the settings on the OTF update panels.
  • Page 105 Table 7–3: OTF update panel – Sequence Settings (continued) Setting or Purpose control Set Collection Specifies the collection method required for directing fraction Method collection in a run. Click the list to view all of the collection methods defined for the project.
  • Page 106 Table 7–4: OTF update panel – Instrument Method Settings (continued) Setting or Purpose control Total Back For updating isobaric separations only, sets the system Pressure operating pressure, in bars, for the duration of a run. Rule: You cannot use the Total Back Pressure setting here to set the initial pressure of a gradient.

  • Page 107: Updating A Run's Settings From The Open Current Method Or Current Sequence

    Table 7–5: OTF update panel – Collection Method Settings (continued) Setting or Purpose control Collection Vial/ Specifies the fraction collection vial or vessel intended to Vessel collect the fraction. Use to change the collection vial or vessel at the next available peak for the duration of the run.
  • Page 108 To update settings from the current method or sequence: From the OTF update panel or the navigation tree, open the current instrument method, collection method, or sequence. Tip: The tabs for the current methods and sequence are highlighted in yellow. Result: The method or sequence opens in OTF update mode.
  • Page 109 Table 7–6: OTF update options Update Option Purpose category Apply changes For making updates that need only apply to to current run the current injection, or row, in a sequence. Rule: When the sequence includes additional rows, and the run-time counter for a current row expires, the settings defined for the next row in the sequence are reapplied.
  • Page 110 Table 7–6: OTF update options (continued) Update Option Purpose category Sequence Apply changes For adding or deleting rows in the current to current sequence. sequence Tip: This setting is typically used for extending a running sequence or changing its injection volumes. Apply changes For adding or deleting rows in the current to all matching...
  • Page 111 7.3.2.1 Example: extending or shortening a stacked-injection sequence By inserting new rows in a current sequence using the "Insert a row" function, you can quickly copy and add existing rows, to extend the running sequence. Figure 7–3: Adding or deleting rows within a stacked-injection sequence Tip: Changes to the current sequence or method are highlighted in yellow, for easy identification.
  • Page 112 Figure 7–4: Changing the current instrument method from an 8% method to a 6% method 7.3.2.3 Example: changing injection volumes in a stacked-injection sequence By increasing or decreasing the injection volume in all pending rows of the current sequence, you can quickly modify the amount of purified product that you collect. Figure 7–5: Increasing the injection volume in the current sequence from 0.5 mL to 1.0 mL Important:...

  • Page 113: Fraction Collection Optimization

    timing problem because of he reduced time between the stacked peak sets. Consequently, if you know beforehand that you might be spontaneously decreasing injection volumes, in the Sequence Wizard, increase the spacing setting before running the sequence. 7.4 Fraction collection optimization During a purification run, products separated from the dissolved sample matrix are carried to the selected fraction-collection flask by the CO and co-solvent.

  • Page 114: Add Additional Equilibration Time

    To plan for the next run: Configure your system shutdown options so that when the sequence queue empties, the system maintains its current run conditions and washes the injection module. Whenever possible, set up purification sequences so that you complete the run and empty the sequence queue before you must empty fraction bottles, refill or change solvents, and change to a new sample.

  • Page 115: Data Analysis

    Data analysis 8.1 Data-log files ChromScope automatically logs data about each run and stores it in data-log files (*.tta). These files appear in the Project's Data Files folder. If you specified a subfolder naming convention when setting up the project, the data-log files are organized by date, sample name, or user name.

  • Page 116: Setting A Preference For File-Open Mode

    Figure 8–1: Opened data-log file 8.1.2 Setting a preference for file-open mode You can select from three file-open modes (New Tab, Current Tab, or Current Tab Overlay) according to how you want to open and view data-log files. When you select a mode, all data-log files you open remain open in that mode until you select another mode.

  • Page 117: Display Preferences

    To set the file-open mode: From the File menu, click File Open Mode > New Tab, Current Tab, or Current Tab Overlay. Alternative: From the toolbar, click New Tab , Current Tab , or Current Tab Overlay Rules: • For New Tab and Current Tab, the information in the data grid pertains only to the single, opened data-log file.

  • Page 118: Specifying Zooming Options And Controls

    • FID signal • PDA settings (for PDA detector use only) 8.2.1 Specifying zooming options and controls The zooming values determine a displayed chart's viewable boundaries. To specify zooming options and controls: From the Edit menu, click Display Options. Alternative: In the toolbar, click Display Options In the Display Options window, Zooming box, select the option Use Global Zoom, to apply the zooming controls to all plots, or select Use Local Zoom...
  • Page 119 • The peak Information settings affect only the data grid; chart and reports remain unaffected. • If you open the data-log file in Current tab Overlay mode, in addition to the selected, peak-information boxes, the Plot Name box (for example Analog 1) appears.

  • Page 120: Specifying Options For Displaying Run Information

    Table 8–2: Peak information fields according to the file-open mode File open in Current tab mode or File open in Current tab Overlay mode New tab mode Peak Name PeakN Name Peak Number (not displayed) Peak Concentration PeakN Concentration Area/Weight Percent PeakN Area/Weight Percent Volume Percent Volume Percent...
  • Page 121 To display run options: From the Edit menu, click Display Options. Alternative: In the toolbar, click Display Options In the Display Options window, in the Run Information box, select the individual run-information fields that you want to appear in the data grid, or click Select All, to select all of the available run-information fields.

  • Page 122: Specifying Graph Display Options

    Table 8–3: Run information fields for Injection Info (continued) Run Information in Display Options Run Information in Injection Info window Column Temperature Column Temperature Column Name Column Column Port Column Port UV Detector Wavelength Wavelength PDA Starting Wavelength PDA Start Wavelength PDA Stopping Wavelength PDA Stop Wavelength Gradient Start Perc...
  • Page 123 To specify graph display options: From the Edit menu, click Display Options. Alternative: In the toolbar, click Display Options In the Display Options window, in the Graph Display box, select the graph components that you want to appear in the data-log file and its plots, or click Select All, to select all of the available graph components.

  • Page 124: Setting Annotation Display Options And Orientation

    8.2.5 Setting annotation display options and orientation Selected Annotations appear in both charts and reports. Tip: When you assign a FID signal, "Weight" and "Weight Percent" appear in place of "Area" and "Area Percent". To set annotations display options and orientation: From the Edit menu, click Display Options.

  • Page 125: Setting Display Preferences For Pda Options

    To specify overlay options and axis offsets in overlay: From the Edit menu, click Display Options. Alternative: In the toolbar, click Display Options In the Display Options window, in the Overlay Options box, select Actual Overlay, to display your data traces on the actual scale on the y axis, or select Normalized Overlay, to display them on a normalized scale.

  • Page 126: Working With Views

    To specify display preferences for PDA options: From the Edit menu, click Display Options. Alternative: In the toolbar, click Display Options In the Display Options window, PDA Options box, specify options for Wavelength, Time, PDA Time Offset, and 3D Graph Resolution. Click OK, to save your display settings.

  • Page 127: Opening A Single Plot In Expanded View

    8.3.1 Opening a single plot in expanded view To open a single plot in expanded view: From the open data file, select the tab of the plot that you want to expand. From the View menu, click Expand > Current Graph. Alternative: Double-click the file tab to view the expanded graph in a new window.
  • Page 128 Table 8–5: Expanded View toolbar controls Control Description Generates a Quick Report of the selected chromatogram or data-log file in PDF format Print quick report to PDF Generates a Custom Report, based on your Custom Report template, of the selected chromatogram or Print custom report data file in PDF format to PDF...

  • Page 129: Integration

    Table 8–5: Expanded View toolbar controls (continued) Control Description Displays a compressed view of a current Zoom Out chromatogram Selects an area of a current chromatogram to enlarge Zoom Box Resets the view to its original size Undo Zoom Displays the axis coordinate value at a current position Data Cursor Copies the current chromatogram to the Windows Copy to Clipboard...

  • Page 130: Using Auto Integration

    8.4.1 Using auto integration You can apply an integration method's settings to a single chromatogram or all chromatograms of an opened data-log file. To use the auto Integration function: Open the data-log file (in Current Tab or New Tab mode) for which you want to integrate peaks.
  • Page 131 Figure 8–2: Manual integration options Exception: You cannot manually integrate peaks on a file opened in Overlay mode. To manually integrate peaks: Open the data-log file (in Current Tab or New Tab mode) in which you want to integrate peaks. Select the tab of the plot that you want to integrate, and from the View menu, click Expand >...

  • Page 132: Calibration Curves

    8.4.2.1 Options for integrating peaks manually Table 8–6: Options for manually integrating peaks Option Choose Peak Select Choose Peak, and then click in the chart area to select both the start and end points for the peak. The peak is formed between the two selected points.

  • Page 133: Calibration Curve Editor

    Tip: You can simplify the calibration-curve building process by using the Sample List Calibration Curve wizard to easily create sample lists with multiple injections of a sample whose component concentrations vary. Concepts: • A level is data points composed of area, height, and concentration. •...
  • Page 134 Figure 8–3: Example calibration curve window 8.5.1.1 Calibration Curve window controls In the Calibration Curve window, you can do all of the tasks associated with building curves and performing statistical analysis. The following table describes the functions of all of the window's menus and controls. July 28, 2014, 715004623 Rev.
  • Page 135 Table 8–7: Working with the Calibration Curve window Window Menu item or Description component control File menu New Sample Prepares the window for building a new sample- curve collection. Open Sample Displays a list of available sample-curve collections. Close Sample Closes the currently active sample-curve collection.
  • Page 136 Table 8–7: Working with the Calibration Curve window (continued) Window Menu item or Description component control Toolbar Enlarges a portion of the calibration chart within Zoom In the entire chart area. (Click Zoom In repeatedly, to further enlarge the area.) Compresses a portion of the calibration chart Zoom Out area.

  • Page 137: Building Calibration Curves

    Table 8–7: Working with the Calibration Curve window (continued) Window Menu item or Description component control Window controls Add New Curve Opens the Name Curve window, to add a new curve to the sample curve collection. Remove Curve Opens a confirmation window used when removing the active curve from the chart, grid, and the Choose Curve list.
  • Page 138 Later, if necessary, you can edit, remove, or clear curves from the opened collection referring to the table "Calibration Curve window controls". Requirements: • You must have a minimum of three data points to build a valid concentration curve. • If you are adding data points by "Adding peaks to Curve", you must have a data- log file open that contains valid integrated peaks.

  • Page 139: Statistical Analysis

    To manually enter data points for area, height, and concentration, click Add Level Manually, specify the settings in Manual Level window according to the following table, and then click OK. Table 8–9: Add Level Manually window settings Setting How to set Area Enter a numeric area value in the text box.
  • Page 140 view statistics by selecting various calibration formulas that the software applies to the open curve. 8.5.3.1 Extrapolating for area, height, or concentration To extrapolate for area, height, or concentration: In the opened Calibration Curve window, select the curve you want from the Choose Curve list.
  • Page 141 • Quadratic (Ax^2 + Bx + C) • Segments View the statistical information in the Statistical Analysis area of the window. Tips: • Click Stats to view additional statistical information in a separate window. • Click Print Report to print a report listing sample name, curve name, calibration curve plot, and curve statistics, on the default printer.

  • Page 142: Peak Concentration

    8.6 Peak concentration After you build calibration curves for your samples or sample components, you can obtain the concentration of peaks in the chart on the basis of peak names or by using a calibration curve. 8.6.1 Determining concentration according to peak name You can assign names to peaks in the integration method or in the data grid.
  • Page 143 To determine concentration using a calibration curve: Open the data-log file (in Current Tab or New Tab mode). Choose the peaks manually or by means of the software's Auto Integration function, using an integration method. From the Tools menu, click Calibration Curve > Compare Peak to Curve. Alternative: In the opened data-log file's chromatogram, right-click the chart area, and select Compare Peak to Curve.

  • Page 144: Data Management And Reporting

    Data management and reporting 9.1 Data-log files 9.1.1 Importing data-log files You can import data-log files (*.tta) from a defined location into a project's Data Files folder (or subfolder). This operation is useful for importing scouting-run information used to set up the timing parameters for stacked-injection sequences. Rule: You can import data-log files to a subfolder only if a subfolder was created when setting up the project.

  • Page 145: Exporting A Set Of Data-Log Files

    chromatogram and reprocessing it. If the file to be exported includes both analog streams and PDA data, then a separate file is created for each analog channel, PDA total absorbance, and PDA single absorbance. To Export a currently opened data-log file: Open the data-log file.
  • Page 146 • Header on each page: Waters logo and data-file name, with complete path • Footer on each page: Waters Corporation • Chromatogram, or plot of detector signal recorded during the run • General Information: • Instrument method Name • Log Author •...

  • Page 147: Printing A Quick Report To A Pdf File

    • For data-log files opened in Current Tab Overlay mode, the file path and file name in the quick report's header is from the last file opened in overlay mode. • Results for peak weight and peak weight percent appear only when you assign an FID signal.

  • Page 148: Custom Report Templates

    9.3 Custom report templates You can define and save a set of custom report templates for each project. Each report template stores the information used to generate a custom report. This information includes the report's displayed information content, the order of this content, the header and footer layout, and page setup.

  • Page 149: Report Template Editor

    To attach report templates to the data file's current chromatogram or to all chromatograms: Open the data-log file. Tip: If you are attaching the template to a single chromatogram, display the chromatogram in the opened file. From the File menu, click Choose Report Template. Select the report template from the list of available templates.
  • Page 150 Figure 9–1: Example Report Template editor window The Report Template canvas is where you define the actual layout of the report template by dragging and dropping objects from the object navigation tree. You can then use the toolbar controls to manipulate these objects and define the page setup. The following table describes all of the various ways that you can manipulate objects on the report canvas.
  • Page 151 To remove an object Select the object and click Delete . To delete columns in a table, right-click the shaded region above the column header, and click Delete Column. Tip: If the table has only one column, to delete it, you must select it and click Delete To reposition Move the mouse pointer over the object's shaded region, and when...

  • Page 152: Custom Reports

    To format system You format system variables in two steps: variables Set the text format for the text box, as described above, and save your settings. Reopen the Properties window, and select the variables you need. 9.3.3.2 Adding headers and footers You can add footers and headers to your custom template, to display information at the top and bottom of every page on a custom report.

  • Page 153: Previewing A Custom Report

    Depending on what input boxes and components are defined for the report template used for the custom report, the report can comprise these main elements: • Header on each page • Plots: • Chromatogram, showing the recorded detector signal during the run •...

  • Page 154: Printing A Custom Report To A Printer

    To print a custom report to a PDF file: Open the data-log file. Click the tab to display the chromatogram that you want to capture in the report. From the File menu, click Print Custom Report To PDF, define the location in which to save the report in the PDF setting window, and click OK.
  • Page 155 • Specifying frequently used integration settings and report layout for the batch- processed data-log files. • Applying a particular batch-screening method to generate the data and reporting output. When you do so, all of the integration, reporting, and selected data- stream settings come from the selected screening method.
  • Page 156 Requirement: To effectively process single absorbance, you must set a single wavelength in the Batch Processing window. To submit data-log files for batch processing: From the Tools menu, click Batch Process. In the Batch Processing window, identify the location (that is, the system and project) containing the files that you want to process.

  • Page 157: Configuring Auto Report Mail

    Select PDF or Default printer for the batch report. Requirement: If you select PDF, you must enter the file location, and then click Save PDF at. Click Start. Results: • Batch reports of the processed data-log files are printed on a default printer or saved as PDF files.

  • Page 158: System Maintenance

    System maintenance 10.1 Maintenance schedule Refer to this table when planning maintenance operations for the Prep 80q SFC system. When performing service or maintenance tasks, always observe Good Laboratory Practice (GLP) guidelines and your laboratory's standard operating procedures. Table 10–1: Maintenance tasks for the Prep 80q SFC system...
  • Page 159 Table 10–1: Maintenance tasks for the Prep 80q SFC system (continued) Task Component Maintenance interval Service sample Injection module Clean the sample loop after each run. injection module Replace the injection needle or syringe when injection performance fails or degrades. Replace the buffer and sample tubing when injection performance fails or degrades.

  • Page 160: System Shutdown Preparation

    Table 10–1: Maintenance tasks for the Prep 80q SFC system (continued) Task Component Maintenance interval Replace or refill Customer supplied As needed. cylinders or bulk source 10.2 System shutdown preparation You must prepare a system for shutdown before actually shutting it down. To do so, you can program an instrument method, collection method, and sequence to set the conditions for cleaning and flushing the system.

  • Page 161: Shutting Down The System For Long Periods

    Leave the system pressurized. Shut down the chiller unit. 10.2.2 Shutting down the system for long periods Long-term shutdown is appropriate for shutting down the system for several days or longer. To perform a long-term system shutdown: Using a suitable cleaning solvent, remove and clean all fraction collection bottles and lines of residual product.

  • Page 162: Replacing The Co Inlet Filter

    • Dual pump heads with cooling chambers for removing the heat of compression from the pump heads, to keep the CO from converting to a gas as it is drawn into the pump • A rupture-disk safety device, to prevent high-pressure damage to pump components •...

  • Page 163: Replacing The P-50 High-Pressure Pump's Inlet And Outlet Check Valves

    To replace the inlet filter: Shut the valve to the CO supply source, and then open the bleeder valve, to completely depressurize the CO pump. Locate the CO inlet filter assembly where the CO supply is plumbed to the system. Figure 10–1: CO inlet filter assembly Inlet filter cover...
  • Page 164 The outlet check valves, atop the pump heads, allow flow out of the pump heads as the piston advances. As the piston retracts, the outlet check valves prevent system pressure from entering the pump heads. When a check valve fails, the pump's outflow decreases, causing the pump to run faster to maintain the specified flow rate.

  • Page 165: Replacing Piston Seals On The P-50 High-Pressure Pump

    Using a 1/2-inch wrench, tighten the check-valve nut approximately 120°. Reinstall the tubing. 10. If using the P-50 to pump liquid CO to the system, open the valve to the CO supply source, and inspect for leaks. 10.3.3 Replacing piston seals on the P-50 high-pressure pump The P-50 high-pressure pump's seals incorporate low-friction sapphire.
  • Page 166 Figure 10–3: Pump head assembly removed Sapphire piston Clean the sapphire piston using acetone and a lint-free cloth. Using the plastic, hooked tool from the service kit, remove the backup bearing and piston seal from the pump head. Figure 10–4: Piston spring seal and backup seal Using a cotton swab, clean the seal cavity using acetone.
  • Page 167 Figure 10–5: Pump head with installed spring seal Slide the backup bearing into the head until it is seats flush. Orient the head so that the two bleed holes on its side are visible when both heads are installed. Figure 10–6: Bleed holes on side of pump head 10.

  • Page 168: High-Pressure Pump

    13. Reinstall the tubing to the check-valve nuts and the manifold. 14. If using the P-50 pump to deliver liquid CO to the system, reinstall the cooling lines attached to the front of the pump heads. Open the valve to the CO supply source, and inspect for leaks.

  • Page 169: Replacing The Rupture Disk

    Clean any remaining pieces of O-ring from the head cavity. Install the replacement O-ring seal in the cavity, and then press the face plate into the head. Loosely install the 4 head nuts between the head and the pump chassis. Begin tightening the nuts, approximately 1 or 2 turns each, in a clockwise pattern, until all are tight.

  • Page 170: Co-Solvent Delivery Pump And Components Maintenance

    To replace a burst rupture disk: Using a 3/4-inch wrench, loosen and remove the disc retainer nut from the manifold. Remove the burst rupture disk. Clean the retainer nut threads and spray with a fluoropolymer dry lubricant. Install the replacement disk into the disc holder with the dome side facing inward.

  • Page 171: Prep Injection Module Maintenance

    CO flow before entering the injection module. For details on maintaining the 2545 QGM and co-solvents, refer to the Waters 2535/2545/2555 Quaternary Gradient Module Operator's Guide. 10.5 Prep injection module maintenance The sample prep injection module injects one sample at a time using a standard 2- mL loop and 5-mL syringe.

  • Page 172: Replacing The Injection Components

    10.5.1.1 Confirming that the loop is free of contamination Run a basic sequence that performs an injection of wash solvent. Monitor the detector plot, to establish whether contamination is present. If contamination is detected, repeat the steps above, to clean the sample loop and syringe.

  • Page 173: 2489 Uv/Visible Detector Maintenance

    10.6 2489 UV/visible detector maintenance The Waters 2489 UV/Visible detector is a 2-channel, ultraviolet/visible (UV/Vis) detector designed for SFC and HPLC applications. The detector operates under ChromScope control and includes a standalone console, for initial detector setup and troubleshooting functions.
  • Page 174 Required materials: Collection module preventive maintenance kit To replace the valve rotor seal and stator assembly: Stop all flow to the unit, and remove the inlet and outlet lines to the valve after noting their connection positions. Figure 10–11: Valve rotor seal and stator assembly Stator screws Stator Stator face assembly...

  • Page 175: Cleaning The Collection Bottles And Flow Lines

    Mount the new rotor seal, with the grooves facing the stator. Tip: The three pins on the shaft assembly fit only one way in the mating holes in the rotor seal. Figure 10–12: New rotor seal mounting orientation Stator mating holes Shaft assembly pins Mount the stator ring on the valve body, and secure it with the stator ring screws.

  • Page 176: Abpr And Heat-Exchanger Module Maintenance

    Thus fuses protect both operator and equipment. Open fuses are properly an extremely rare occurrence. Fuses that regularly open indicate a probable fault in the component and must be investigated. Contact Waters Customer Support for assistance.
  • Page 177 Notice: To avoid damaging system modules, before replacing an opened fuse, determine the cause of the fuse's failure. It is possible that a serious underlying problem requires attention. Contact Waters Customer Support for assistance. Warning: To avoid electrical fire, ensure that replacement fuses comply with the ratings affixed to the rear panel of the module.

  • Page 178: Consumables And Spare Parts

    Close the fuse-holder door, reconnect the power cord, and move the power switch to the "on" (1) position. 10.10 Consumables and spare parts To ensure that your system operates as designed, use only Waters Quality Parts. Visit www.waters.com/wqp for information about Waters Quality Parts, including how to order them.

  • Page 179: Diagnostic Tests And Troubleshooting

    Exit the ChromScope software, and cycle power to the failed module. Restart the ChromScope software. If the alarm persists, report its occurrence, along with any supplemental information, to your Waters service representative. July 28, 2014, 715004623 Rev. A Page 179...

  • Page 180: Internal Hardware Or Software Alarm Shut-Down System

    Restart the ChromScope software. If the alarm persists, report its occurrence, along with any supplemental information, to your Waters service representative. 11.2 Reviewing the history log ChromScope automatically logs history records for each session and stores them in the Histories Folders node.

  • Page 181: Pressure Testing The System

    Figure 11–1: Example history folder records 11.3 Pressure testing the system Pressure test the system when any of these events occur: • The system or one of its components is newly installed. • The system is recommissioned after being shut down for a week or more. •...

  • Page 182: Troubleshooting System Problems

    At the inlet of the co-solvent reservoir, use a graduated cylinder to measure the volume of co-solvent drawn into the co-solvent pump and compare that volume with the volume exiting the system. Important: Always measure the co-solvent pump's flow rate from the inlet of the co-solvent reservoir using a graduated cylinder.

  • Page 183: Troubleshooting The P-Series High-Pressure Pumps

    Defective check valve Replace the CO pump's check valve. Leaking piston seal Replace the CO pump's piston seal. Defective flow meter Contact your Waters technical service representative. July 28, 2014, 715004623 Rev. A Page 183...

  • Page 184: Pressure Reading Reported On Display Fails To Match That Of Other System Components

    Ensure valves are in proper positions, inspect for valves proper tubing sizes and properly cut tubing, and confirm valve alignments. Defective pressure Contact your Waters technical service representative. transducer Miscalibrated pressure Contact your Waters technical service representative. transducer 11.5.3 Repeated failures of pump check valves Table 11–3: Repeated failures of pump check valves...

  • Page 185: Troubleshooting Heat Exchanger Problems

    Investigator) or heat exchanger is equipped with an internal temperature safety switch that was activated, owing to a problem with the controller or heater element. Call a Waters service representative. Zone is not enabled on Verify that the correct zone is enabled in the software's temperature controller Heater Controller panel.

  • Page 186: Troubleshooting System Pressure Problems

    • Replace the module's fuse. • The model of ABPR module is equipped with an internal temperature safety switch that was activated, owing to a problem with the module. Call a Waters service representative. Leaks in the system, Inspect the system for leaks or problems associated with or problems with pump output.

  • Page 187: A Safety Advisories

    Heed all warnings when you install, repair, or operate any Waters instrument or device. Waters accepts no liability in cases of injury or property damage resulting from the failure of individuals to comply with any safety precaution when installing, repairing, or operating any of its instruments or devices.

  • Page 188: Specific Warnings

    Warning: (Risk of explosion.) A.1.1 Specific warnings A.1.1.1 Burst warning This warning applies to Waters instruments and devices fitted with nonmetallic tubing. Warning: To avoid injury from bursting, nonmetallic tubing, heed these precautions when working in the vicinity of such tubing when it is pressurized: •...
  • Page 189 A.1.1.2 Biohazard warning The following warning applies to Waters instruments and devices that can process material containing biohazards, which are substances that contain biological agents capable of producing harmful effects in humans. Warning: To avoid infection with potentially infectious, human-sourced products, inactivated microorganisms, and other biological materials, assume that all biological fluids that you handle are infectious.

  • Page 190: Notices

    Notice: To avoid damaging the instrument’s case, do not clean it with abrasives or solvents. A.3 Warnings that apply to all Waters instruments and devices When operating this device, follow standard quality-control procedures and the equipment guidelines in this section.
  • Page 191 警告: 規制機関から明確な承認を受けずに本装置の変更や改造を行うと、本装置のユーザーとして の承認が無効になる可能性があります。 Warning: Use caution when working with any polymer tubing under pressure: • Always wear eye protection when near pressurized polymer tubing. • Extinguish all nearby flames. • Do not use tubing that has been severely stressed or kinked. •...
  • Page 192 Avvertenza: fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione: • Indossare sempre occhiali da lavoro protettivi nei pressi di tubi di polimero pressurizzati. • Spegnere tutte le fiamme vive nell'ambiente circostante. • Non utilizzare tubi eccessivamente logorati o piegati. •...
  • Page 193 • 非金属チューブには、テトラヒドロフラン(THF)や高濃度の硝酸または硫酸などを流さないでくださ い。 • 塩化メチレンやジメチルスルホキシドは、非金属チューブの膨張を引き起こす場合があり、その場 合、チューブは極めて低い圧力で破裂します。 This warning applies to Waters instruments fitted with nonmetallic tubing. This warning applies to instruments operated with flammable solvents. Warning: The user shall be made aware that if the equipment is used in a manner not specified by the manufacturer, the protection provided by the equipment may be impaired.

  • Page 194: Electrical Symbols

    Avvertenza: si rende noto all'utente che l'eventuale utilizzo dell'apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall'apparecchiatura. Advertencia: el usuario deberá saber que si el equipo se utiliza de forma distinta a la especificada por el fabricante, las medidas de protección del equipo podrían ser insuficientes.

  • Page 195: Handling Symbols

    A.5 Handling symbols The following handling symbols and their associated statements can appear on labels affixed to the packaging in which instruments, devices, and component parts are shipped. Symbol Description Keep upright! Keep dry! Fragile! Use no hooks! July 28, 2014, 715004623 Rev. A Page 195...

  • Page 196: B Specifications

    Specifications B.1 Prep 80q SFC system specifications The Waters Prep 80q SFC system is a bulk-scale, preparative system designed to serve as a high-pressure separator for quantitative recovery of purified products, such as enantiomers, complex synthetic chemicals, and natural products.

  • Page 197: Mass Flow Meter Specifications

    B.4 Column specifications The standard column supplied with the Prep 80q SFC system and used to validate the system during installation and maintenance is a Viridis Silica 2-EP OBD column, 5 µm, 19 × 150 mm.

  • Page 198: Automated Back-Pressure Regulator Specifications

    B.7 Automated back-pressure regulator specifications The ABPR valve assembly is motor-driven and temperature controlled, to compensate for cooling during depressurization. A built-in pressure sensor provides closed-loop feedback for control and pressure-alarm monitoring. B.8 Collection-bottle cabinet specifications A collection cabinet houses six, safety-coated 2-L bottles, and one 20-L HDPE bottle, for waste collection.

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