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Dionex UltiMate 3000 Series Operating Instructions Manual

Fluorescence detectors
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UltiMate 3000 Series
Fluorescence Detectors
FLD-3100 and FLD-3400RS
Operating Instructions
Revision: 1.0
Date: November 2009
© 2009 Dionex
Doc. No. 4820.7850

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  • Page 1 UltiMate 3000 Series Fluorescence Detectors FLD-3100 and FLD-3400RS Operating Instructions Revision: 1.0 Date: November 2009 © 2009 Dionex Doc. No. 4820.7850...
  • Page 2 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Operating Instructions...

  • Page 3: Declaration Of Conformity

    Product: Detector Types: FLD-3100 and FLD-3400RS Dionex GmbH herewith declares conformity of the above products with the respective requirements of the following regulations:  Low-Voltage Directive 2006/95/EC  EMC Directive 2004/108/EC The electrical safety of the products was evaluated based on the following standard: ...
  • Page 4 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page II Operating Instructions...

  • Page 5: Table Of Contents

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Table of Contents Introduction........................1 How to Use This Manual ..................1 Safety Information ....................2 1.2.1 Symbols on the Instrument and in the Manual ..........2 1.2.2 General Safety Precautions................3 Intended Use ......................
  • Page 6 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Preparation for Operation (Startup) ................33 Overview......................33 Connecting the Drain System ................33 Installing the Flow Cell..................34 Equilibrating the System..................36 General Guidelines for Detector Operation ............38 4.5.1 Mobile Phases ....................38 4.5.2 Mobile Phase Delivery System................39 4.5.3...
  • Page 7 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Monitoring System Functions................72 5.8.1 Wavelength Calibration and Verification ............72 5.8.2 Recording the Flow Cell Temperature............. 73 5.8.3 Predictive Performance..................74 5.8.4 Operational Qualification and Performance Qualification ......74 Shutting Down the Detector ................74 5.10...
  • Page 8 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page iv Operating Instructions...

  • Page 9: Introduction

    The layout of this manual is designed to provide quick reference to the sections of interest to the reader. However, in order to obtain a full understanding of the detector, Dionex recommends that you review the manual thoroughly before beginning operation.

  • Page 10: Safety Information

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 1.2 Safety Information .2 Safety Information The CE Mark label on the instrument indicates that the detector is compliant with the The CE Mark label on the instrument indicates that the detector is compliant with the related standards (...

  • Page 11: General Safety Precautions

     When the panels are removed, dangerous electrical connections will be exposed. Disconnect the detector from all power sources before removing the panels.  Always replace blown fuses with original spare part fuses from Dionex ( page 91).  Replace faulty power cords and communication cables.

  • Page 12: Consignes Générales De Sécurité

     Before interrupting operation for several days or more or when preparing the detector for transport, observe the precautions for shutting down the detector ( page 74).  Use original Dionex spare parts only. Substituting non-Dionex parts or using non-Dionex accessories may impair the performance of the instrument.
  • Page 13  Remplacez toujours les fusibles grillés par des fusibles de rechange d'origine Dionex. ( page 74).  Remplacez les cordons d'alimentation électrique et les câbles de communication défectueux.

  • Page 14: Intended Use

    91).  Utilisez des pièces de rechange d'origine Dionex. Effectuer des remplacements par des pièces ne provenant pas de Dionex ou utiliser des accessoires ne provenant pas de Dionex peut affecter les performances de l'instrument.  N'utilisez pas l'instrument de manière autre que celles décrites dans ce manuel.

  • Page 15: Federal Communications Commission (Fcc) Note

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 1.4 Federal Communications Commission (FCC) Note This equipment has been tested and found to comply with the limits for a Class A digital device, pursuant to part 15 of the U.S. FCC Rules. These limits are designed to provide reasonable protection against harmful interference when the equipment is operated in a commercial environment.
  • Page 16 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page 8 Operating Instructions...

  • Page 17: Overview

    2 Overview 2.1 Unit Description The detector is a high-quality instrument designed for HPLC analysis as part of a Dionex UltiMate 3000 system. The module was developed especially for HPLC and UHPLC analysis and can be used in numerous laboratory environments for routine analysis and sophisticated HPLC research tasks: ...

  • Page 18: Operating Principle

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.2 Operating Principle .2 Operating Principle Fluorescence detectors are optical detectors. In a fluorescence detector, the sample is Fluorescence detectors are optical detectors. In a fluorescence detector, the sample is exposed to light at a defined wavelength (excitation). The thus excited sample substance exposed to light at a defined wavelength (excitation).
  • Page 19 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS The excitation light stimulates the sample to emit fluorescence light. When the light exits the flow cell, it is focused by the emission optics (no. 6) through the emission monochromator (no. 8).

  • Page 20: Configurations

    ( no. 10, Fig. 3, page 11). Depending on the PMT setting, the instrument selects the suitable PMT for each measurement, or you can manually select which PMT should be used. For emission scans, you can scan with both PMTs simultaneously. The installation of the additional PMT is performed by Dionex Service. Description Part No.

  • Page 21: Interior Components

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.4 Interior Components .4 Interior Components The front panel door tilts downward to provide easy access to the inside front panel, for The front panel door tilts downward to provide easy access to the inside front panel, for example, for maintenance and repair work.

  • Page 22: Front Panel Elements

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.5 Front Panel Elements .5 Front Panel Elements Fig. 5: Front panel view Front Panel Function Element Display Shows information about the detector: - General information upon power up ( page 41) - Status screen, depending on the operating mode (page 41)

  • Page 23: Rear Panel

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.6 Rear Panel Fig. 6: Rear panel Description Power switch ( page 15) Fuse cartridge ( page 16) Main power receptacle ( page 24) Type label USB 2.0 (Universal Serial Bus) port for connection to the Chromeleon computer (...

  • Page 24: Fuse Cartridge

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.6.2 Fuse Cartridge The fuse cartridge contains two slow-blow fuses, rated at 2 A (5 x 20 mm). For information about how to change the fuses, see page 91. 2.6.3 USB Port The Chromeleon Chromatography Management System uses a USB connection to control the detector.

  • Page 25: Fluid Connections

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.7 Fluid Connections The front panel door tilts downward to provide easy access to the fluid connections in the detector. At dedicated positions in the front part of the enclosure, six slots are provided for the capillaries: on the top and bottom of the enclosure (two slots each), and on the left and right side of the enclosure (one slot each) (...

  • Page 26: Lamp

    In addition, you can extend the lamp lifetime by selecting a different lamp mode ( page 70). The lamp must be replaced only by Dionex Service. 2.10 Leak Sensor A leak sensor ( Fig. 4, page 13) is installed inside the detector. If liquid collects in the drip tray under the fluid connections, the leak sensor reports a leak, and the Status LED on the front panel door changes to red.

  • Page 27: Detector Control

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.11 Detector Control The detector can be controlled by the Chromeleon Chromatography Management System. To operate the detector with Chromeleon, an appropriate Chromeleon version and the corresponding license are required. Tip: All software details in this manual refer to Chromeleon 6.80.

  • Page 28: System Wellness, Predictive Performance, And Diagnostics

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 2.12 System Wellness, Predictive Performance, and Diagnostics System Wellness monitors the health of the detector. Therefore, the detector supports several performance and reliability features that can help you detect small problems before they turn into big ones: ...

  • Page 29: Installation

    Tips: Immediately report any shipping damage to both, the incoming carrier and Dionex. Shipping insurance will compensate for the damage only if reported immediately. Keep the original shipping container and packing material. They provide excellent protection for the instrument in case of future transit.

  • Page 30: Positioning The Detector In The Ultimate 3000 System

    3.3 Positioning the Detector in the UltiMate 3000 System If the detector is part of an UltiMate 3000 system, for example, for analytical HPLC applications, Dionex recommends stacking the individual modules. However, the arrangement of the system modules depends on the application.

  • Page 31: Connecting The Detector

    Connect the detector directly to the USB port on the computer.  Connect the detector to an internal USB port on the pump. Dionex recommends connecting all modules to the pump, and then connecting the system to the Chromeleon computer via only one connection ( Fig. 8, page 24). If the system includes a UV detector in addition to the fluorescence detector, Dionex recommends connecting the UV detector directly to the Chromeleon computer.

  • Page 32: Connecting The Power Cord

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Solvent Rack Pump Autosampler Column compartment Fluorescence detector Fig. 8: Example for the rear panel connections on an UltiMate 3000 system 3.4.3 Connecting the Power Cord Use the power cord shipped with the detector to connect the instrument to the main power source.

  • Page 33: Connecting The Analog Outputs (Optional)

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 3.4.4 Connecting the Analog Outputs (Optional) 3.4.4.1 Installing the DAC Module 1. Turn off the detector and disconnect the power cord from its source. 2. Use appropriate grounding protection to avoid electrostatic discharge, which may result in damage to electronic components.

  • Page 34: Connecting An Analog Output To An Evaluation Device

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 3.4.4.2 Connecting an Analog Output to an Evaluation Device 1. Locate the analog output cable (part no. 6074.0002) provided with the DAC board. Plug the cable connector into the analog output on the detector rear panel.

  • Page 35: Setting Up The Detector In Chromeleon

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 3.5 Setting Up the Detector in Chromeleon This section provides brief instructions for setting up the detector in Chromeleon. For details, see the Chromeleon Help. Tip: When the detector is connected to the Chromeleon computer, verify that the Chromeleon software is installed before turning on the detector power for the first time.
  • Page 36 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 7. Windows Vista will automatically detect the new detector and perform the USB installation. If Windows Vista fails to detect the detector and launches a wizard instead, this indicates that you connected the detector to the computer and turned on the power for the first time before you installed Chromeleon.

  • Page 37: Installing The Detector

    5. Open the Add device to timebase dialog box. To do so, click Add Device on the Edit menu or right-click the timebase and click Add Device on the menu. 6. On the Manufacturers list, click Dionex HPLC: UltiMate 3000 and on the Devices list, click FLD-3x00(RS) Detector.

  • Page 38: Virtual Mode

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS General Page Shows the general instrument parameters. Fig. 11: General page  Virtual Mode Verify that the virtual mode is set to off. If the virtual mode is enabled, the Module Address box will be unavailable. If you exit this page without having entered a module address, the virtual mode will be enabled automatically.

  • Page 39: Device Name

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS  Download Click this button to update the detector firmware with the version available in Chromeleon. (The button appears dimmed if the virtual mode is enabled.) The detector is shipped with the most recent firmware version. If a firmware update is ever required, follow the steps in section 7.6 (...

  • Page 40: Changing The Configuration Properties

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Signals Page The page lists all signal channels that the detector can record. A name is defined for each channel. To allow raw data collection for a channel, select the Enabled check box next to the channel name.

  • Page 41: Preparation For Operation (Startup)

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 4 Preparation for Operation (Startup) 4.1 Overview After you have unpacked, positioned and connected the detector as described in sections 3.1 through 3.5 ( page 21 and following), prepare the detector for operation. Follow this sequence of steps: 1.

  • Page 42: Installing The Flow Cell

    40 bar (micro flow cell). Do not connect any capillaries or instruments to the flow cell out line if this might generate a higher backpressure. Dionex recommends using a capillary or tube with an internal diameter of at least 0.5 mm at the flow cell out line.
  • Page 43 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 1. Remove the cover on the flow cell opening (if installed). To do so, loosen the two finger-tight screws. No tools are required to loosen the screws. The screws are captive in the cover and do not need to be removed.

  • Page 44: Equilibrating The System

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 3. Tighten the flow cell screws hand-tight. Flow cell screws Fig. 18: Tighten the flow cell screws 4. Remove the dummy plugs on the flow cell in and out connection ports. Connect the flow cell inlet to the separation column and connect a waste capillary to the flow cell outlet.
  • Page 45 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Perform system equilibration in Chromeleon or select the required commands and parameters on the front panel menus of the instruments. To equilibrate the system from Chromeleon  Select and perform the operating commands and parameters from the Commands dialog box.

  • Page 46: General Guidelines For Detector Operation

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 4.5 General Guidelines for Detector Operation The following sections offer general guidelines for detector operation. For information about how to optimize the detector performance, see section 5.7 ( page 62). 4.5.1 Mobile Phases Mobile phase quality significantly affects detection limits and instrument performance.

  • Page 47: Mobile Phase Delivery System

    The band spring should be stainless steel, a fluoropolymer (Kalrez or fluoro- silicone), or other material of known quality. If you are working with an UltiMate 3000 system, using original Dionex parts only, all the above recommendations are met.
  • Page 48 ( Fig. 19). The flow cell must be shipped in its original packaging. Fig. 19: Original Dionex flow cell plugs  During longer idle times, the flow cell should not be filled with pure water to avoid the growth of algae.

  • Page 49: Operation And Maintenance

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5 Operation and Maintenance The detector can be controlled by the Chromeleon Chromatography Management System. For details, see section 5.4 ( page 50) and the following. 5.1 Power-Up To start the detector for the first time, turn on the main power switch on the rear panel of the detector.
  • Page 50 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Monitoring the progress of data acquisition on the detector display After data acquisition was started via Chromeleon (AcqOn), you can watch the progress of data acquisition on the display. The display shows a scaled fluorescence value (in counts) at the selected wavelength.

  • Page 51: Menus On The Detector Display

    Four white spots can be found on the front panel below the display. To show the menu key, select the position of the utmost left spot on the detector display with the Dionex menu pen select the position of the utmost left spot on the detector display with the Dionex menu pen (part no.

  • Page 52: Detector Menus

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.3.1 Detector Menus Fig. 24 shows an overview of the detector menus. For information about the general menu layout and structure, see page 45. For information about the individual commands and parameters, see sections 5.3.3 through 5.3.5 ( page 47 and following pages).

  • Page 53: General Menu Layout And Structure

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.3.2 General Menu Layout and Structure In general, the menu layout is as follows: > Main (1..3) 1. Control 2. Diagnostics 3. Service   Select Back Fig. 25: Menu layout (here Main menu) Description Reports the menu name and the number of items on the menu list.
  • Page 54 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS To … Select … - Return to the previous menu level. Back - Return to the status screen (from the Main menu). Toggle between two operating states (for example, between Silent and Audible in the Toggle Leak sensor mode).

  • Page 55: Main Menu

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.3.3 Main Menu The Main menu provides top-level access to the menu structure. To open the Main menu, show the function keys and select Menu ( page 43). > Main (1..3) 1. Configuration 2.

  • Page 56: Configuration Menu

    Fig. 27: Configuration menu To … Select … Reset all important detector settings to the Dionex standard settings. In Reset to factory defaults the Reset to factory defaults? dialog box, select OK to confirm the restore or select Cancel to keep your settings.

  • Page 57: Diagnostics Menu

    View information about the flow cell, such as, serial number, flow cell type, and Flow Cell flow cell material. View information about the temperatures inside the detector Temperatures 5.3.6 Service Menu The Service menu provides information for the Dionex service when exchanging a lamp. Operating Instructions Page 49...

  • Page 58: Chromeleon Software

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.4 Chromeleon Software Before you begin, verify that 1. The Chromeleon software is installed on the computer and the license code is entered. 2. The detector is connected to the Chromeleon computer via a USB connection.

  • Page 59: Direct Control

    To open the Commands dialog box for the detector 1. Open a control panel (any panel is possible). To open a control panel, open the Chromeleon Browser and double-click a control panel in the Dionex Templates/Panels folder. 2. Connect the control panel to the timebase in which the detector is installed. On the Control menu, select Connect to Timebase, and then select the timebase on the Timebase tab.
  • Page 60 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Fig. 29: Commands dialog box 6. Verify that the detector is connected to Chromeleon. If it is not, select Connect to connect the detector. For a list of the commands and properties that are supported for the detector, see the Chromeleon Help.

  • Page 61: Automated Control

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Fig. 30: Detector control panel (here FLD-3400RS) The control panel provides access to the operating parameters and commands required for routine operation of the detector. Additional functions are available in the Commands dialog box.
  • Page 62 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS First, select whether you want to record one or several emission channels. Select the parameters for the channel or channels in the table. For information about parameter settings, see page 62. The wizard page changes depending on whether you select one or...
  • Page 63 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS When you record multiple channels (FLD-3400RS only), the baseline noise increases at higher data collection rates. The maximum data collection rate and response time depend on a number of parameters and are always automatically calculated on the basis of the selected parameters.
  • Page 64 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS To create a program manually 1. Open an existing program. Select and double-click the program you want to open. - or - On the File menu, select Open. In the dialog box, select Program on the Object of Type list and select the program.

  • Page 65: Smartstartup And Smartshutdown

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS To start a program Program for sample analysis 1. Create a sample list (sequence). A sequence must include the program and a method for evaluating the sample data (for example, for peak identification, area determination, and amount determination).

  • Page 66: Operational Settings

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.5 Operational Settings 5.5.1 Detecting Liquid Leaks in the Detector Leak detection is enabled as a standard when the detector is shipped. When leak detection is active and the leak sensor reports a leak ...

  • Page 67: Operational Modes Of The Detector

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.6 Operational Modes of the Detector 5.6.1 Single-Channel Mode A fluorescence detector is usually operated in single-channel mode, that is, a single excitation/emission wavelength pair is measured. As this mode provides the highest possible data rates, always use the single-channel mode for very small peaks.

  • Page 68: Spectra Scanning

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.6.4 Spectra Scanning With the FLD-3x00(RS) detectors, you can also record several types of spectra. The excitation monochromator or emission monochromator (or both) moves over a settable wavelength range, while the intensity of the fluorescence signals is measured and recorded continuously for each wavelength.
  • Page 69 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS To perform a spectra scan Tips: During the scan process, no other commands can be sent to the fluorescence detector. The duration of the scan process depends on the range to be scanned and the selected scan speed.

  • Page 70: Optimizing Detector Performance

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.7 Optimizing Detector Performance The performance of the detector can be optimized by careful selection of key operating parameters. The table summarizes these parameters, indicates the performance characteristics affected, and offers guidelines for selecting the parameters.
  • Page 71 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Method using 1 sample with the fluorescence detector 1. Set the excitation wavelength to a wavelength at which most sample components absorb. Many fluorescent molecules absorb at 250 nm. Set the filter wheel setting to a wavelength that is at least 30 nm above the excitation wavelength (in the example 280 nm).
  • Page 72 Fig. 34: 3D data recorded with a diode array detector Wavelength switching in Chromeleon Dionex recommends using the single-channel mode to record data by switching the wavelengths in-between the detected peaks of the various sample components, rather than to simultaneously measure all interesting wavelengths in multi-channel mode.

  • Page 73: Sensitivity (Detector Sensitivity)

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.7.2 Sensitivity (Detector Sensitivity) The sensitivity is used to optimize the signal-to-noise ratio in a chromatogram. Depending on the intensity of the light signal, it may be necessary to adjust the sensitivity several times during an analysis.
  • Page 74 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Determining the optimum Sensitivity Determine the optimum sensitivity in a separate sample run after you have determined the optimum wavelengths ( page 62). Use a sample (standard) with the maximum expected analyte concentration.

  • Page 75: Filter Wheel Setting (Fld-3400Rs Only)

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Tips: MaxPMTSaturation affects the signals of both PMTs and the signals of all active channels (FLD-3400RS only) . You can completely turn off fluorescence detection by setting the sensitivity to Off. This may be required, for example, if you want to record the incoming signal from a UV detector for a sample of high concentration.

  • Page 76: Response Time And Data Collection Rate

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS The FLD-3400RS provides a filter wheel that is located before the emission monochromator. The filter wheel carries several optical filters with different cut-off lengths. The qualitative transmission behavior of all filters is shown in Fig. 36, with a transmission range of approx.

  • Page 77: Multi-Channel Performance

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Response Time  Select a response that is about 1/3 of the peak width at half-height of the narrowest peak of interest. A longer response time allows more averaging of the signal and results in less short-term noise.

  • Page 78: Flow Cell Temperature

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.7.6 Flow Cell Temperature The temperature of a sample influences its fluorescence. Thus, a problem with fluorescence detection is that the results can strongly be influenced by fluctuations in ambient temperature. The FLD allows you to heat the flow cell and thus keep temperatures inside the flow cell at a constant level.

  • Page 79: Baseline Behavior

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Tip: You can change the lamp mode during a measurement, for example, from LongLife mode (between the peaks) to Standard or HighPower mode (for the interesting peaks). This extends the lamp lifetime, without any loss in sensitivity in the important areas.

  • Page 80: Monitoring System Functions

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 5.8 Monitoring System Functions This section provides a short overview of some special functions for qualification, documentation and monitoring the system that Chromeleon supports for the detector. Some functions are available also on the control panel for the detector. For additional information about a function, see the Chromeleon Help.

  • Page 81: Wavelength Calibration

    Commands dialog box for the detector and on the FLD Options page in the Program Wizard ( page 53). Dionex recommends always recording the flow cell temperature. If a problem occurs, the temperature channel can provide helpful information to identify and eliminate the source for the problem.

  • Page 82: Predictive Performance

    Operational Qualification and Performance Qualification allow you to check and document the performance of the HPLC system. All materials required for performing qualification and detailed instructions are available from Dionex on request. 5.9 Shutting Down the Detector Observe the following precautions before interrupting the operation or before shipping the detector: ...
  • Page 83 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS If you are running Chromeleon, you can set the detector and HPLC system into the standby mode or automate system shutdown, as described in the sections below. Standby Program A standby program sets the HPLC system into standby mode. The application can be reactivated very quickly afterward.

  • Page 84: Routine And Preventive Maintenance Intervals

    Check the volume of the liquid in the waste container and empty as needed. Monitor the lamp ( page 86). Annually Have a Dionex service representative check the detector once a year to prevent contamination, excessive wear, etc. Tip: Chromeleon supports functions for monitoring qualification and service intervals (...

  • Page 85: Troubleshooting

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 6 Troubleshooting 6.1 Overview The following features help you to identify and eliminate the source for problems that may occur during the operation of the detector or UltiMate 3000 system. Status Screens The status indicators on the front panel provide a quick visual check of the operational status of the detector.

  • Page 86: Messages On The Detector Display

    The following table lists the most important detector-related messages along with recommended remedial actions. In addition to the messages in the table, other messages may appear. In this case, please note the exact wording of the message and contact Dionex Service if you are unable to eliminate the problem.
  • Page 87 Enter a different start and end time for the wavelength scan range. start and end position. Service interval expired. The service interval has expired. Have Dionex Service perform system maintenance. Service interval warning - The service interval will expire after the number of days indicated in the % days remaining.
  • Page 88 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS When the detector is operated with Chromeleon and if communication between Chromeleon and the detector cannot be established, related messages may appear in the Chromeleon Audit Trail. Message Remedial Action The USB connection between the detector and the Chromeleon server FLD-3100@USB-1610103 - may be interrupted.

  • Page 89: Operating Problems

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 6.3 Operating Problems The following table provides information about common operating problems that might occur with an UltiMate 3000 system and lists probable causes, as well as remedial actions. For more information, also see the manuals for the other modules of an UltiMate 3000 system.
  • Page 90 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Problem Probable Cause Remedial Action High noise level, There are pressure fluctuations Purge the pump; check general pulsation / from the pump. function ( Pump Manual). The Raman S/N value was not reached.
  • Page 91 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Problem Probable Cause Remedial Action Poor reproducibility The autosampler draws air from There is not enough amount of the vial. sample in the vial, the needle height setting is incorrect ( Autosampler Manual), or there are too many replicates.
  • Page 92 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Problem Probable Cause Remedial Action Poor reproducibility The sample is unstable and Use new sample or change the (Cont'd) decomposes. conditions. The environmental conditions are Make sure that the temperature and unstable.

  • Page 93: Service

    For more information, see the warranty statement in the terms of sale.  Before you return any instrument to Dionex for repair, contact Dionex Service or your local distributor. An RMA (Return Material Authorization) number is required in order to track your instrument.

  • Page 94: Lamp

    Fluorescence Detectors FLD-3100 and FLD-3400RS 7.2 Lamp The lamp must be replaced only by Dionex Service. Never try to exchange the lamp or open the enclosure yourself. You can monitor the lamp age. This function can help to decide when a lamp is due to be replaced.

  • Page 95: Flow Cell

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 7.3 Flow Cell 7.3.1 Flushing the Flow Cell Flush the flow cell when you suspect that eluted compounds may have deposited on the cell walls. Complete the following steps: 1. Flush the flow cell with HPLC-grade water.

  • Page 96: Removing The Flow Cell

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS  Capillary connections between the column and flow cell should be as short as possible to avoid peak broadening effects due to excessive dead volume.  The backpressure on the flow cell must not exceed 20 bar (analytical flow cell) or 40 bar (micro flow cell).

  • Page 97: Installing The Flow Cell

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 4. Remove the flow cell from the optical bench. Tip: You can flush the flow cell by using the optional syringe injection/flushing kit (part no. 6078.4200). 7.3.2.2 Installing the Flow Cell 1. Insert the flow cell straight into the cell opening.

  • Page 98: Drying The Leak Sensor

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 7.4 Drying the Leak Sensor .4 Drying the Leak Sensor The leak sensor is installed inside the detector and reports a leak when liquid collects in The leak sensor is installed inside the detector and reports a leak when liquid collects in the drip tray under the fluid connections.

  • Page 99: Replacing The Main Power Fuses

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 7.5 Replacing the Main Power Fuses .5 Replacing the Main Power Fuses Warning: Turn off the main power switch and disconnect the power cord from its source. Avertissement: Avant de remplacer les fusibles, arrêtez le détecteur. Assurez-vous de bien débrancher le cordon d'alimentation de la source secteur.

  • Page 100: Updating The Detector Firmware

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 7.6 Updating the Detector Firmware The detector is shipped with the most recent firmware version. The detector firmware is also included in Chromeleon. To check which firmware version is installed in the detector and which version is included in Chromeleon: ...
  • Page 101 If the download is not successful, the related messages appear in the Audit Trail. In this case, turn off the detector. Turn on the detector again and repeat the download as described above. If the download fails, contact Dionex Service. Operating Instructions...
  • Page 102 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page 94 Operating Instructions...

  • Page 103: Technical Information

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 8 Technical Information Specification FLD-3100 FLD-3400RS Optical Design: Two monochromators with concave holographic gratings and hyperbolic mirrors for highest efficiency in light transmission Light source: Xenon flash lamp (15W); Three different flash frequencies (HighPower, Standard, LongLife) can be selected.
  • Page 104 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Control: All parameters and functions software controlled, USB 1.1 3 LEDs (Power, Connected, Status) for status monitoring In Chromeleon: GLP features: Full support of automatic equipment qualification (AutoQ) and system performance monitoring.

  • Page 105: Accessories, Consumables, And Spare Parts

    Open-end wrench (size 3/16") 6146.1231 Dionex menu pen 6300.0100 USB cable type A to type B, 5m 6911.0002 * The part number always refers to the packing unit. For more information, contact your Dionex sales representative. Operating Instructions Page 97...

  • Page 106: Optional Accessories

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 9.2 Optional Accessories Description Part no. Remarks Analytical flow cell 6078.4230 For installation details, see page 34. (cell volume: 8 µL, cell material: steel, capillary ID: 0.25 mm, pressure limit: 20 bar) Micro flow cell Availability upon request.

  • Page 107: Consumables And Spare Parts

    USB cable, type A to type B, high speed USB 2.0 (cable length: 5 m) 6911.0002 * The part number always refers to the packing unit. For more information, contact your Dionex sales representative. ** Availability upon request. Operating Instructions...
  • Page 108 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page 100 Operating Instructions...

  • Page 109: Appendix: Common Mobile Phases

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 10 Appendix: Common Mobile Phases The mobile phase composition affects its UV cutoff, that is, the minimum effective wave- length for the measurement. In general, mobile phases are solvents, such as, water, acetronitrile, methanol, or other substances.
  • Page 110 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page 102 Operating Instructions...

  • Page 111: Index

    UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS 11 Index Commands Dialog Box ........52 Configuration ............ 49 Configurations........... 11 2nd PMT ............31 Connection Analog Output ..........24 Chromeleon Computer........22 Accessories Fluid.............. 16 Optional ............103 Power Cord........... 23 Standard ............102 USB ..............
  • Page 112 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Excitation Monochromator........9 Excitation Scan ..........61 Lamp ............17, 91 Excitation Wavelength ........64 Lamp Mode ..........72 Replace ............91 Lamp Age............91 Filter Wheel ............69 Leak Detection ..........59 Firmware Download ........30, 97 Leak Sensor...........
  • Page 113 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Optics..............9 Service Menu............. 50 Optimization ............64 Shut Down............77 Optimum Wavelength........64 Shutdown Program..........77 Signals (Dialog Box) ......... 32 Single-Channel Mode........60 SmartShutdown ........... 58, 78 Panel Tabset............53 SmartStartup..........38, 58 Performance Optimization.........64 Soft Keys ............
  • Page 114 UltiMate 3000 Series: Fluorescence Detectors FLD-3100 and FLD-3400RS Page 106 Operating Instructions...

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