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PerkinElmer 200A Series User Manual

Fluorescence detector
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LIQUID CHROMATOGRAPHY
S
200
ERIES
A
F
D
LUORESCENCE
ETECTOR
User's Guide

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Table of Contents

Troubleshooting

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Summary of Contents for PerkinElmer 200A Series

  • Page 1 LIQUID CHROMATOGRAPHY ERIES LUORESCENCE ETECTOR User’s Guide...
  • Page 2 The information contained in this document is subject to change without notice. Except as specifically set forth in its terms and conditions of sale, PerkinElmer makes no warranty of any kind with regard to this document, including, but not limited to, the implied warranties of merchantability and fitness for a particular purpose.

  • Page 3: Table Of Contents

    Environmental Conditions ..............28 Electrical Safety ..................29 EMC Compliance ................... 30 Europe ..................30 United States ................. 30 WEEE Instructions for PerkinElmer Products..........31 Warning Labels on the Instrument ............32 System Description .................35 Overview ..................... 37 Control Switches and Indicators............38 Keyboard on the UI Pad (Option).............
  • Page 4 Environmental Conditions for An Installation Location......59 Checking the Contents..............59 Assembling ..................60 Removal of Transport Screws ............60 Tubing Arrangements ..............61 UI Pad................... 62 Spare Parts Available ................65 Operation ..................67 General Information about Entering Parameters ........71 Setup ....................

  • Page 5: Introduction

    Introduction...

  • Page 7: Note

    2. No part of this manual may be reproduced, transmitted or disclosed to a third party in any form or by any means without the express written permission of PerkinElmer, Inc. 3. PerkinElmer, Inc. assumes no liability for any direct, indirect, or consequential damages arising from use not described in this manual.

  • Page 8: Product Warranty

    8 . Series 200a User’s Guide Product Warranty This product is warranted to be free from defects in material or workmanship under normal use within the product specifications indicated in this manual and under conditions given below. This warranty is void if the instrument is not used according to the instruction manual. The manufacturer makes no warranties, either express or implied, except as provided herein, including without limitation thereof, warranties as to marketability, merchantability, for a particular purpose or use, or against infringement of any patent.
  • Page 9 Introduction g. Failure due to maintenance or repair by other than service personnel qualified by the manufacturer. h. Failure due to relocation or transport conducted not under the supervision of the manufacturer after the initial installation of the instrument. Failure due to disassembly, modification or relocation not approved by the manufacturer.

  • Page 10: About This Manual

    About This Manual This user’s guide is divided into following chapters: Chapter 1 Introduction This chapter contains the product warranty, conventions used in the manual and the conventions and warnings used in the manual. Chapter 2 Safety Practices Important safety information is provided in this chapter. Chapter 3 System Description This chapter contains information on the detector, a descriptions of Fluorometry and instrument specifications.

  • Page 11: Conventions Used In This Manual

    UPPERCASE text, for example, ENTER or ALT, refers to keys on the PC keyboard. '+' is used to show that you have to press two keys at the same time, for example, ALT+F. All eight digit numbers are PerkinElmer part numbers unless stated otherwise. Notes, cautions and warnings Three terms, in the following standard formats, are also used to highlight special circumstances and warnings.
  • Page 12 12 . Series 200a User’s Guide We use the term CAUTION to inform you about situations that could CAUTION result in serious damage to the instrument or other equipment. Details about these circumstances are in a box like this one. Caution (Achtung) Bedeutet, daß...
  • Page 13 Introduction . 13 We use the term WARNING to inform you about situations that could result in personal injury to yourself or other persons. Details about these circumstances are in a box like this one. WARNING Warning (Warnung) Bedeutet, daß es bei Nichtbeachten der genannten Anweisung zu einer Verletzung des Benutzers kommen kann.

  • Page 14: Warning Signs On The Instrument

    14 . Series 200a User’s Guide Warning Signs on the Instrument WARNING FOR CONTINUED PROTECTION REPLACE ONLY WITH SAME FUSE TYPE AND RATING Power On/Off push button Alternating current...

  • Page 15: Safety Practices

    Safety Practices...

  • Page 18: Overview

    This advice is intended to supplement, not supersede, the normal safety codes in the user's country. It is also a supplement to the PerkinElmer standard Safety and Health Policy. The information provided does not cover every safety procedure that should be practiced. Ultimately, maintenance of a safe laboratory environment is the responsibility of the analyst and the analyst's organization.

  • Page 19: Common Safety Precautions

    If any item is missing or damaged or if you have any questions, please notify PerkinElmer. NOTE:...

  • Page 20: Ignition Of Flammable Chemicals

    • If any warning/caution label has become illegible due to deterioration with age or it has been damaged due to any cause, contact your PerkinElmer service representative. We use the term WARNING to inform you about situations that could result in personal injury to yourself or other persons. Be sure that all instrument operators read and understand the precautions listed below.

  • Page 21: Electric Shock Due To Contact With Inside Of Instrument

    Safety Practices . Electric Shock Due to Contact with Inside of Instrument • The xenon lamp is supplied with high voltage of 30 kV, which could cause an electric shock to result in serious or fatal injury. Before removing the light source cover for replacement of the light source lamp, be sure to turn off the power switch and unplug the power cord before proceeding.

  • Page 22: Lamp Installation

    Series 200a Fi User’s Guide Be sure to wear proper protective gear such as safety goggles, safety mask, thick long sleeves, and gloves when handling the xenon lamp. WARNING Lamp Installation • When loosening or tightening the retaining nut for the xenon lamp, be careful not to apply excessive force to its glass bulb part.

  • Page 23: Inflammation Or Injury Due To Toxic Or Corrosive Solvent

    Safety Practices . Inflammation or Injury Due to Toxic or Corrosive Solvent When using a toxic or corrosive solvent, be careful not to incur a physical inflammation or injury. For details of the properties of each solvent and how to handle it, refer to the relevant Material Safety Data Sheets (MSDS).

  • Page 24: Precautions On Use Of Solvents

    Series 200a Fi User’s Guide Precautions on Use of Solvents • Fluorocarbon resin and quartz materials are used for the wetted parts of the instrument. Never use solvents that would corrode these materials. • This instrument has internal parts made of materials that could be corroded by strongly acidic solutions, strongly alkaline solutions or organic solvents.

  • Page 25: Disposal Of Waste Solution

    Safety Practices . Disposal of Waste Solution Be sure to collect the waste solvent and treat it for proper disposal in accordance with the relevant laws and regulations regarding water pollution control and sewage treatment. Improper treatment of waste solution may result in environmental pollution and could also lead to a penalty.
  • Page 26 Series 200a Fi User’s Guide We use the term CAUTION to inform you about situations that could CAUTION result in serious damage to the instrument or other equipment. Details about these circumstances are in a box like this one. Be sure that all instrument operators read and understand the precautions listed below.

  • Page 27: General Operating Conditions

    Safety Practices . General Operating Conditions Only use the Series 200a indoors and under the following conditions: Temperature C to 35 Relative Humidity 25% to 80% relative humidity (non-condensing) If possible, avoid any adjustment, maintenance and repair of the opened, operating instrument. If any adjustment, maintenance and repair of the opened instrument are necessary, this must be done by a skilled person who is aware of the hazard involved.

  • Page 28: Environmental Conditions

    Series 200a Fi User’s Guide Environmental Conditions Normal Operating Conditions: • Indoors use. • The location must be free of smoke, dust, corrosive fumes, direct sunlight and excessive vibration. • Ambient temperature: +10°C to +35°C (+50°F to +95°F). • Relative humidity: 25% to 80%, without condensation. •...

  • Page 29: Electrical Safety

    Lethal voltages are present at certain areas within the instrument. Installation and internal maintenance of the instrument should only be performed by a PerkinElmer Service Representative or similarly authorized and trained person. WARNING When the instrument is connected to line power, opening the instrument covers is likely to expose live parts.

  • Page 30: Emc Compliance

    Series 200a Fi User’s Guide EMC Compliance Europe All information concerning EMC standards is in the Declaration of Conformity, and these standards may change as the European Union adds new requirements. United States This product is classified as a digital device used exclusively as industrial, commercial, or medical test equipment.

  • Page 31: Weee Instructions For Perkinelmer Products

    0800 90 66 42 (Monza) Products from other manufacturers may also form a part of your PerkinElmer system. These other producers are directly responsible for the collection and processing of their own waste products under the terms of the WEEE Directive. Please contact these producers directly before discarding any of their products.

  • Page 32: Warning Labels On The Instrument

    Periodically check the appearances of these warning labels to see if they are clean to allow easy reading over a safe distance. If any one of the warning labels becomes illegible due to deterioration, contact PerkinElmer. WARNING...
  • Page 33 Safety Practices . Figure 2-2 Overview of the warning labels on the front of the instrument...
  • Page 34 Series 200a Fi User’s Guide Figure 2-3 Detail of the warning labels on the front of the instrument...

  • Page 35: System Description

    System Description . System Description...
  • Page 36 Series 200a Fi User’s Guide...

  • Page 37: Overview

    Overview The liquid chromatograph system consists of a pump for feeding an eluent, a sample introducing section (autosampler, manual injector) for injecting a sample, a column for chromatographic separation of the injected sample, a column oven for maintaining the column at a constant temperature, and a detector section for detecting a signal of a separated component of the sample.

  • Page 38: Control Switches And Indicators

    Series 200a Fi User’s Guide Functional Description of the Detector Control Switches and Indicators Power Switch UI Pad LEDs Fig. 3-2 Front View Power switch : Turns power ON/OFF. UI pad (option) : A keypad required for unit operation. LEDs : Four LED indicators are provided for indicating the following states (Table 1-1).Doors (upper and lower right/left) : Three doors are provided.

  • Page 39: Keyboard On The Ui Pad (Option)

    System Description Table 3-1 LEDs Designation (Color) LED Indication Green : CH1 operation mode. (green, orange) Orange : An error has occurred in CH1 operation mode. Green : CH2 operation mode. (green, orange) Orange : An error has occurred in CH2 operation mode. Xe LAMP (green) Green : Xe lamp is lit.
  • Page 40 Series 200a Fi User’s Guide Table 3-2 Indication Function Remarks (Auto zero) See Chapter 5 Sets the absorbance to zero at the indicated wavelength. Operation. (Wavelength) See Chapter 5 Used to set the wavelength of monochromator. Operation. (Utility = Set up) SET UP 1.
  • Page 41 System Description (cont’d) Indication Function Remarks Accesses the previous screen. Accesses the next screen. (Start) See Chapter 5 START 1. Starts the time program. Operation. 2. Starts analog output of the stored spectrum. (Stop) See Chapter 5 STOP 1. Stops the time program. Operation.

  • Page 42: The Rear Panel

    Series 200a Fi User’s Guide The Rear Panel 2 Lamp 4 Marker In Off In (option) 1 E-Line 5 Recorder (option) 6 Processor 3 Auto (option) Zero In Fig. 3-4 Rear Panel * Explanation of each function is given in Table 3-3.
  • Page 43 System Description Table 3-3 Indication Function Remarks LAMP OFF IN The lamp is turned off by means of a contact signal input. AUTO ZERO IN The auto zero function is activated by means of contact signal input. MARKER IN The marker function is activated by The marker function is started (option) means of contact signal input.

  • Page 44: Description Of Fluorometry

    Series 200a Fi User’s Guide Description of Fluorometry Principle of Fluorometry Excitation Fluorescence or Phosphorescence Stable condition Unstable condition (Ground state) (Excited state) Radiationless Radiationless transition transition Excited state V = Excited triplet state Light Light Excitation light Ground state V = Absorption Fluorescence Phosphorescence...

  • Page 45: Advantages Of Fluorometry

    System Description Figure 3-5 shows the energy level transitions of an organic molecule for the various processes involved in absorption, fluorescence and phosphorescence. When light strikes an organic molecule in the ground state, it absorbs radiation of specific wavelengths and several excited states are populated. A part of the excitation (absorbed) energy is lost in vibrational relaxation, i.e.
  • Page 46 Series 200a Fi User’s Guide In this discussion, we will consider the detection of a sample that has a transmittance of 99% (relative to the blank). If we assume that the inaccuracy of the % transmittance measurement is 0.1%, the reliability of the observed data is as follows: Percent transmittance of blank 100.0 ±...

  • Page 47: Notes On Fluorescence Analysis

    System Description In addition to providing high sensitivity, fluorescence detection can provide a fluorescence spectrum and an excitation spectrum (which is very similar to the absorbance spectrum). These may be useful in identifying the eluent. If the eluent contains two compounds, selection of the appropriate excitation and emission wavelengths may be used to provide qualitative and quantitative information about the components in the mixture.
  • Page 48 Series 200a Fi User’s Guide The Rayleigh peak and the Raman peak will occur even if the eluent does not contain any compound that fluoresces. If there is any doubt whether an observed peak is a Raman peak, a Rayleigh peak or a peak due to the fluorescence from the compound of interest, simply change the excitation wavelength slightly.
  • Page 49 System Description When the concentration of the compound of interest is relatively high, the concentration reported by fluorescence detection may be lower than the actual concentration. This phenomenon is due to the "Inner Filter effect" which is depicted in Fig. 3-9. In this example, the concentration of the compound of interest is sufficiently high whereby molecules that are close to the entrance slit absorb essentially all of the excitation radiation.
  • Page 50 Series 200a Fi User’s Guide 3. Second-order and Third-order Radiation A diffraction grating that is set to transmit radiation of a specific wavelength (e.g. 250 nm) will also transmit multiples of the radiation (e.g. 500 nm). When a fluorescence spectrum is collected, the analyst should be aware of the second order (and third order) phenomena and take care that peaks are not misidentified.

  • Page 51: Principle Of Operation

    System Description Principle of Operation Diffusion plate Excitation Plane mirror monochromator Sample Monitor detector (Flow cell) (Photodiode) Xenon Measuring lamp detector Emission (Photomultiplier) monochromator Xenon lamp (Fluorescence starter monochromator) Xenon lamp Photomultiplier ballast high voltage circuit Monitor detector Sample amplifier fluorescence amplifier Operational...

  • Page 52: Features Of Light Source

    Series 200a Fi User’s Guide Features of Light Source The light source thermally dissociates ozone that is formed from the ultraviolet radiation via the heat generated by the xenon lamp itself. This allows the use of a fused-silica xenon lamp that produces intense radiation even in the short wavelength region.

  • Page 53: Specifications

    System Description Specifications 1. Optical system: Ratio photometry; intensity of transmitted beam is monitored. 2. Light source: Xe lamp (150 W) Hg lamp (for checking wavelength) 3. Measuring wavelength range Excitation wavelength: 200 to 850 nm/Zero order 4. Measuring wavelength range emission wavelength: 250 to 900 nm/Zero order Photomultiplier must be changed at emission wavelength greater than 731 nm.
  • Page 54 Series 200a Fi User’s Guide 16. Spectrum memory: Memorized contents Excitation and fluorescence spectra Number of spectra memorized Measuring wavelength range Changeable in measurement condition setting 17. Marker (option): Marker output of approx. 0.6 mV at recorder terminal (corresponding to about 6 graduations on chart) 18.

  • Page 55: Installation

    Installation Installation...
  • Page 56 Series 200a Fi User’s Guide...

  • Page 57: Unpacking

    The customer must not attempt installation of the instrument at delivery. To insure safe and correct use NOTE: of the instrument, its installation must be carried out your PerkinElmer service representative. Before installation of this instrument the customer shall make preparations for satisfying the installation requirements in accordance with this manual.

  • Page 58: Installation Conditions

    Series 200a Fi User’s Guide Installation Conditions For installation of this instrument, confirm the following conditions. 1. Power Supply Power voltage: 100 to 115/220 to 240 V AC Fluctuation Within ±10% of the rated voltage Frequency: 50 or 60 Hz Fluctuation Within ±0.5 Hz of the rated frequency Power consumption:...

  • Page 59: Environmental Conditions For An Installation Location

    (e.g., stirrer, vibrator). Checking the Contents After unpacking, check the delivered contents against the packing list furnished with the instrument. If anything is missing or damaged, or if you have any question, please contact PerkinElmer. Contents: User’s Guide on CD...

  • Page 60: Assembling

    Series 200a Fi User’s Guide Assembling Removal of Transport Screws 1. Remove three screws from the bottom face. Use a Phillips screwdriver to remove transport screws, *A, *B and *C Transport screw *C (for lamp house) Transport screw *B (for Transport screw *A main body) (for main body)

  • Page 61: Tubing Arrangements

    Installation After removing transport screws, store them in a safe place so you will not lose them. NOTE: Fig. 4-2 Transport Screw Removal/Xe Lamp Attachment Tubing Arrangements In the Series 200a, provide inlet tubing to the pump and waste solution tubing through the right piping section of the drain reservoir.

  • Page 62: Ui Pad

    Series 200a Fi User’s Guide UI Pad This section describes how to use the UI pad for operations. The UI pad is provided with common control keys available for different modules and unique control keys for particular modules. The functions of the common control keys are described below. 1.
  • Page 63 Installation (cont’d) Function Designation CL (clear) Clears an input numeric value during input (after a numeric value or the decimal point is entered; but before the ENT key is pressed). The cursor is indicated at the top of the input part. The previous input value is presented as a preset value.
  • Page 64 Series 200a Fi User’s Guide ii. Input of plural numeric values: Parameter name Input item LAMP CHANGE (MM, DD, YYYY) <1 - 12> Input part Input range iii. Selection of a numeric value: Selectable items Parameter name TIME CONSTANT (0.1s=1, 0.5s=2, 2.0s=3, 4.0s=4, 8.0s=5) Input part iv.

  • Page 65: Spare Parts Available

    Installation Spare Parts Available The fuse and xenon lamp are the only replacement parts and consumables for this instrument. It is suggested that you stock a few fuses and lamps in the laboratory to minimize down time. Replacement Parts Part No. Description 0999-1667 Fuse - 5 Amp 250 V Slo-Blow...
  • Page 66 Series 200a Fi User’s Guide...

  • Page 67: Operation

    Operation Operation...
  • Page 68 Series 200a Fi User’s Guide...
  • Page 69 Operation Turning on the Power Power Switch UI Pad Fig. 5-1 Front View of the Series 200a If the monitor display screen does not appear on the LCD within 3 minutes after turning on the power, or NOTE: if an error message appears on the LCD, refer to ‘Troubleshooting. 1.
  • Page 70 Series 200a Fi User’s Guide The sequence of wavelength drive (check of reference point for drive), lighting of the lamp and auto zeroing is performed. 3. When the above steps are successfully completed, the 1st monitor screen is presented. Accesses the 2nd monitor screen. , ESC (escape): Returns to the 1st monitor screen.

  • Page 71: General Information About Entering Parameters

    Operation General Information about Entering Parameters 1. Entering a value, press numeric key then ENT (enter) key. 2. Erasing the present data entry a. Made an error in entering data, but have NOT pressed ENT key, press CL (clear) key, then re-edit the value.

  • Page 72: Setup

    Series 200a Fi User’s Guide Setup Before analytical operation, communication channel (CH) and liquid crystal display contrast (LCD CONTRAST) need to be set. • This setting is not required every time. When unnecessary, skip this setting. • The default value or currently set value is indicated before your input. Selecting the Communication Channel (CH) Set a communication channel via the E-Line in the following way.

  • Page 73: Setting The Liquid Crystal Display Contrast

    Operation <Setup initial screen> SETUP: DETECTOR CONTRAST Serial No. When you press the ENT key here, the setup items selecting screen is presented to allow reentry. Or you can advance to LCD contrast setting. The selected setting will be fetched upon turning on power supply at the next time. 6.

  • Page 74: Setting Of Standard Xenon Lamp Lifetime

    Series 200a Fi User’s Guide Injury due to Xenon Lamp Explosion Replace the xenon lamp with a new one before its cumulative turn-on time exceeds the guaranteed useful lifetime. With the lapse of the guaranteed useful lifetime, the bulb wall darkens due WARNING to evaporation of electrode material and adhesion of scattered substances, resulting in a decrease in heat dissipation.
  • Page 75 Operation SETUP ITEMS (DETECTOR=1, CONTRAST=2 EXCHANGE STANDARD=3) Select EXCHANGE STANDARD. Press numeric key 3 and then ENT key. 4. The following display will be presented. <Standard Xe lamp lifetime setting screen> XE LAMP EXCHANGE STANDARD (1 - 2500) H _500 The standard lifetime of the Xe lamp is settable here.

  • Page 76: Operating Instructions

    Series 200a Fi User’s Guide Operating Instructions Make sure the power switch of the detector is ON, and the display presents the 1st monitor screen. • Then the unit should be allowed to warm up at least 1 hour for stabilization of the xenon lamp and accuracy of measurement.

  • Page 77: Setting The Utility

    Operation Pressing the key enables you to advance to the emission wavelength setting without changing the excitation wavelength. Then, the cursor moves to Em. 3. Set an emission wavelength (Em). <Em wavelength setting screen> (250 - 900.0) nm Press numeric keys (to input 2 5 0 to 9 0 0) and then ENT key. This wavelength is settable within a range from 250 to 900 nm in increments of 1 nm.
  • Page 78 Series 200a Fi User’s Guide UTILITY (OFFSET=1, TIME CONSTANT=2, USE PROG=3, PMT VOLT=4, OTHERS=5) Select OFFSET. Press numeric key 1 and then ENT key. c. The following display will be presented. <Offset setting screen> OFFSET (0 - 1000) Set an offset value. Press numeric key (0 to 1 0 0 0, usually input 0) and then ENT key.
  • Page 79 Operation 2. Setting the TIME CONSTANT Setting item: Electrical response time a. Press the UTLTY key. b. The following display will be presented. <Utility initial screen> UTILITY (OFFSET=1, TIME CONSTANT=2, USE PROG=3, PMT VOLT=4, OTHERS=5) Select TIME CONSTANT. Press numeric key 2 and then ENT key. c.
  • Page 80 Series 200a Fi User’s Guide Press numeric key (1 or 0) and then ENT key. 1: Time program starts by start input. 0: Time program does not start despite start input. Default value: 1 d. Selection or disable of a time program is now completed and the 1st monitor screen returns. When use of a time program is disabled, “-”...
  • Page 81 Operation Press the UTLTY key. ii. The following display will be presented. <Utility initial screen> UTILITY (OFFSET=1, TIME CONSTANT=2, USE PROG=3, PMT VOLT=4, OTHERS=5) Select OTHERS. Press numeric key 5 and then ENT key. iii. The following display will be presented. <Others setting screen>...
  • Page 82 Series 200a Fi User’s Guide Select OTHERS. Press numeric key 5 and then ENT key. iii. The following display will be presented. <Others setting screen> OTHERS (Em BANDWIDTH=1, ERROR OUT=2, LEAK SENSOR=3, LAMP=4) Select ERROR OUT. Press numeric key 2 and then ENT key. iv.
  • Page 83 Operation LEAK SENSOR (OFF=0, ON=1) You can select use (1) or no-use (0) of the leak sensor. Press numeric key (0 or 1, usually input 1) and then ENT key. Default value: 0 When use (1) of the leak sensor is selected, an error message will be indicated upon detection of liquid leakage.

  • Page 84: Setting The Glp Function

    Series 200a Fi User’s Guide <1st monitor screen> TIME PROG LAMP The Xe lamp comes on when this unit is powered on. Shortly after xenon lamp is turned off, it cannot be turned on again, since it is still at a high temperature. NOTE: Therefore, measurement is impossible.
  • Page 85 Operation <Logbook selecting screen> LOGBOOK (REPORT=1, RESET=2) Select Report. Press numeric key 1 and then ENT key. e. On the logbook report screen, the total ON period of the Xe lamp and its standard lifetime (in parentheses under Xe LAMP), the number of switching-on times and the date of the last lamp change will be indicated.
  • Page 86 Series 200a Fi User’s Guide Press numeric key 2 and then ENT key. Next, advance to the Xe lamp change date setting screen for setting the date of xenon lamp change. e. Set the date of xenon lamp change in the following procedure. i.
  • Page 87 Operation Press numeric key 1 and then ENT key. c. The following display will be presented. <Lamp selecting screen> LAMP SELECT (Xe=1, Hg=2) Select Hg. Press numeric key 2 and then ENT key. d. The following display will be presented. <Logbook selecting screen>...
  • Page 88 Series 200a Fi User’s Guide c. The following display will be presented. <Lamp selecting screen> LAMP SELECT (Xe=1, Hg=2) Select Hg. Press numeric key 2 and then ENT key. d. The following display will be presented. <Logbook selecting screen> LOGBOOK（REPORT=1, RESET=2） Select RESET.
  • Page 89 Operation Logbook setting is now completed and the 1st monitor screen returns. When a mercury lamp change date is set, the total ON period of the Hg lamp and the number of switching-on times will be automatically reset accordingly. 5. Setting of KEY LOCK Keys can be locked for prevention of wrong operations.
  • Page 90 Series 200a Fi User’s Guide c. The WL accuracy checking screen appears and it is checked whether wavelength is normal or not. <WL accuracy checking screen (running)> CHECKING WL ACCURACY (Hg) If an Hg lamp lighting error occurs, the error message shown below will appear. Return to the 1st monitor screen by pressing the CL key.

  • Page 91: Spectrum Measurement

    Operation 7. CHECK (Xe lamp energy check) Energy level of the xenon lamp can be checked using the xenon lamp beam at 254 nm. a. Press the GLP key. b. The following display will be presented. <GLP initial screen> (LOGBOOK=1, KEY LOCK=2, WL CHECK=3, Xe CHECK=4) Select Xe CHECK.
  • Page 92 Series 200a Fi User’s Guide The spectral bandwidth is 15 nm on both excitation and emission sides with the Series 200a FL detector. NOTE: A component with a difference between excitation and emission wavelengths of less than 30 nm cannot be accurately measured.
  • Page 93 Operation Press numeric keys 2 5 0 to 8 5 0 and then ENT. Input value Value of less than 850 within WL1 + 50 to WL1 + 400. Setting range 250 to 850 nm Initial (default) value: Pressing key prior to ENT key will return to WL1. e.
  • Page 94 Series 200a Fi User’s Guide SPECTRUM MODE (FL (Ex)=1, FL (Em)=2) Press numeric key 2 and then ENT to select FL (Em). Now proceed to the scan range setting screen for setting a scan range, and enter a start wavelength (cursor will appear at WL1). c.

  • Page 95: Setting Of Recorder And Spectrum Output Factors

    Operation Initial (default) value: blank Upon entering a memorize spectrum No., the previous spectral data will be deleted and the new data will be memorized. When the Xe lamp goes off, press the ENT key and the 1st monitor screen will return.
  • Page 96 Series 200a Fi User’s Guide RECORD (RCD RANGE=1, RCD SPEED=2, SPECT RCD=3) Press numeric key 1 and then ENT to select RCD RANGE (recorder range). c. The recorder full scale range setting screen appears. <Recorder full scale range setting screen> RCD RANGE (1 - 1000) 1000 Press numeric keys 1 to 1 0 0 0 and then ENT to set a recorder full scale range.
  • Page 97 Operation b. The recorder setting screen appears. <Recorder setting screen> RECORD (RCD RANGE=1, RCD SPEED=2, SPECT RCD=3) Press numeric key 3 and then ENT to select SPECT RCD (spectral recording). c. The output data spectrum No. setting screen appears. <Output data spectrum No. setting screen> RECORD SPECTRUM NO.

  • Page 98: Operation Of Marker

    Series 200a Fi User’s Guide <Error display> SPECTRA ARE MISMATCHED PRESS “CL” KEY TO CLEAR MESSAGE e. The “ready for output” monitor screen will appear. <“Ready for output” monitor screen> RCD NO. SPEED WL-RANGE (Em) WIDTH:STD 200-600 PRESS “START” Press the START key. Pressing the ESC key prior to START key will cancel the process.
  • Page 99 Operation Up to 9 programs can be registered as a time program, and a total of 100 steps can be registered for the 9 programs. The registered time program is retained by battery backup even when the power switch is turned OFF. Use the CL key to correct entered numerics.
  • Page 100 Series 200a Fi User’s Guide 100. 0: Conditions in the time program No. selected above are used; additions or changes can be made in the program. 1: Conditions in the time program No. selected above are deleted, and new conditions can be set. The system now advances to the step setting screen for setting the time program, and setting is allowed for the first step (the cursor moves to TIME).
  • Page 101 Operation Press numeric keys 2 5 0 to 9 0 0 and then ENT key. Setting range: 250 to 900 nm It is recommended to set the same wavelength as was set for Em of the measuring wavelengths. Initial (default) value: blank The cursor next moves to BASE.
  • Page 102 Series 200a Fi User’s Guide 102. Press numeric keys 0 to 6 0 0 and then ENT key. Setting range: 0 to 600 minutes, in increments of 0.1 minute The cursor now moves to Ex. ii. Set an Ex (excitation) wavelength to be changed. <Step setting screen (Ex indicated)>...
  • Page 103 Operation vi. When the setting for the second step is completed, return to the TIME setting on step setting screen. If more parameters will be changed, then repeat the operation in this subsection. If parameters will not be changed, advance to the final step setting. Setting of final step of time program: Set the measurement end TIME.
  • Page 104 Series 200a Fi User’s Guide 104. Described next is the procedure for newly creating a time program shown in the figure. HOLD Auto zero START STOP TIME (minutes) 10.0 20.0 Ex (nm) Em (nm) Base- line Described here is an example of newly setting a time program at program No. 1. Ten minutes after the start (1), photomultiplier voltage is changed from MID (3) to LOW (4), excitation wavelength from 250 to 400 nm, emission wavelength from 350 to 500 nm, and hold baseline function is applied, and then measurement is completed after 20 minutes (2).

  • Page 105: Edit The Wavelength Program

    Operation Edit the Wavelength Program If you need to edit the wavelength program follow the next procedure. Press PROG key. ↓ SET PROGE NO. (1 ~ 9) Press 1 key and then ENT. ↓ EDIT MODE (MODIFY=0, NEW=1) Press 1 key and then ENT. ↓...
  • Page 106 Series 200a Fi User’s Guide 106. Press 2 key and then ENT. ↓ TIME BASE 10.0 Press 2 0 keys and then ENT and ESC keys. ↓ TIME BASE 20.0 Press the ESC key. Time program setting is now finished. ↓...
  • Page 107 Operation Press key to access the next step. ↓ TIME BASE 10.0 Press key to access the next step. ↓ TIME BASE 20.0 Press the ESC key, and the 1st monitor screen returns. ↓ TIME PROG LAMP...

  • Page 108: Shutdown Procedure

    Shutdown Procedure Turn OFF the detector’s power switch to shut it down. The set measurement conditions are retained by the memory even when the power is switched off. When power is turned on again, the conditions that were in place when power was turned off will be restored.

  • Page 109: Flushing The Flow Cell After Use

    Operation Flushing the Flow Cell After Use Make sure to flush the flow cell with distilled water after using any buffers or salts in the mobile phase. Failure to carry out flushing may lead to clogging and/or breakage of the flow cell.

  • Page 110: Before Performing Analysis

    Series 200a Fl User’s Guide 110. Before Performing Analysis This chapter describes the basic items to be careful about before starting analysis. Ignition of Flammable Chemicals 1. This instrument is not explosion-proof. In unattended operation, do not use organic solvents having an ignition point below 70 °C. WARNING 2.
  • Page 111 Operation Inflammation or Injury due to Toxic or Corrosive Solvent When using a toxic or corrosive solvent, be careful not to incur a physical inflammation or injury. WARNING For details of the properties of each solvent and how to handle it, refer to the relevant Material Safety Data Sheets (MSDS).

  • Page 112: Characteristics Of Organic Solvents

    Series 200a Fl User’s Guide 112. Table 5-1 Reagents for Mobile Phase that may be Restricted Use is Use is Possible if Use is Possible if 10% or Reagents Unsuitable for Possible 50% or Lower Lower SUS316 Phosphoric Acetic acid Disodium phosphate Ammonium chloride acid...
  • Page 113 Operation Ignition of Flammable Chemicals b. This instrument is not explosion-proof. In unattended operation, do not use organic solvents having an ignition point below 70 °C. WARNING c. Beware of ignition hazard when using flammable chemicals such as organic solvents. a.
  • Page 114 Series 200a Fl User’s Guide 114. Table 5-2 Characteristics of Organic Solvents Ignition Point Vapor Specific Polarity Viscosity Refractive UV Cut Flash Boiling Density Gravity (cP20 °C) Lower Upper Index off (nm) Point (°C) Point (°C) °C (Air: 1) (Water: 1) Limit Limit Fluoroalkanes...
  • Page 115 Operation Warnings for Using Organic Solvents as Mobile Phase 1. When using organic solvents, be careful about UV cut off wavelength. WARNING UV cut off wavelength Light cannot be transmitted in the range shorter than this wavelength. For example: Cut off wavelength of chloroform is 245 nm. Chloroform cannot be used in the range shorter than 245 nm (e.g.

  • Page 116: Cautions On Static Electricity

    Series 200a Fl User’s Guide 116. Cautions on Static Electricity Ignition of Flammable Chemicals by Static Electricity When using flammable chemicals, be careful about possible ignition due to static electricity. To prevent the build-up of static electricity, use a WARNING conductive container for waste solution and provide proper grounding connection to it.

  • Page 117: Maintenance

    Maintenance Maintenance...
  • Page 118 Series 200a Fl User’s Guide 118.

  • Page 119: Operational Check

    Operational Check Check the operation when the instrument has been installed. 1. Prepare the instrument for operation (see chapter, Operation) 2. Turn ON the power switch of the detector. 3. Check that measurement conditions (time constant, output range, etc.) can be properly set. 4.

  • Page 120: Performance Check

    Series 200a Fl User’s Guide 120. Performance Check Check the performance when the instrument has been installed. NOTE: First adjust the xenon lamp position and then check the performance. Positional adjustment of the lamp is necessary for an accurate performance check. When this detector unit is used in combination with the EZChrom Elite Client/Server, the lamp NOTE: energy, wavelength accuracy and sensitivity can be automatically calculated.
  • Page 121 Maintenance . 121 Series 200 Autosampler 3 x 3 Column In-Line Solvent Filter (Optional) Series 200 LC Pump Solvent Systems Kit Series 200a Fluorescence Detector Fig. 6-1 Tubing Connection to Place Water in the Flow Cell 2. Measurement of Raman Spectrum of Water Set the flow cell unit in place, measure the Raman spectrum of water under the conditions given below, and record the results (see chapter 5, Operation ).
  • Page 122 Series 200a Fl User’s Guide 122. iii. The 1st monitor screen appears again. Set a time constant and a photomultiplier voltage. Press the UTLTY key, and the utility setting screen appears. Press numeric key 2 and then ENT to select time constant. ii.
  • Page 123 Maintenance . 123 viii. The other setting screen appears. Press numeric key 1 and then ENT to select Em BANDWIDTH. ix. The Em bandwidth setting screen appears. Press numeric key 1 or 2 and then ENT to set an emission spectrum bandwidth. STANDARD= emission spectrum bandwidth 15 nm WIDE=emission spectrum bandwidth 30 nm.
  • Page 124 Series 200a Fl User’s Guide 124. Press numeric keys 4 5 0 and ENT to set WL2. iii. The memorize spectrum No. setting screen now appears. Press a numeric key (1 to 4) and ENT to select MEMORIZE SPECTRUM NO. iv.
  • Page 125 Maintenance . 125 v. The recorder speed setting screen will appear. Press numeric key 2 and then ENT to select 60 nm/min. vi. The 1st monitor screen returns. vii. Press the RECORD key. The recorder setting screen appears again. Press numeric key 3 and then ENT to select SPECT RCD. viii.
  • Page 126 Series 200a Fl User’s Guide 126. • A spectrum is recorded by starting the recorder or chromato-data processor simultaneously with pressing the START key. xi. The spectrum output screen appears, and spectrum output begins. xii. When the spectrum output is completed, the 1st monitor screen returns automatically.
  • Page 127 Maintenance . 127 In measurement with the emission spectrum bandwidth set at WIDE (30 nm), the peak emission NOTE: wavelength is shifted toward the shorter wavelength side from 397 nm. Due to the WIDE setting, the spectrum bandwidth widens for the scattered light of excitation beam and the Ramen line of water.
  • Page 128 Series 200a Fl User’s Guide 128. 7. Measurement of Raman Spectrum of Water Using a Time Program (when Raman signal intensity obtained in the preceding subsection (3) is low). If the sensitivity (S/N ratio of Raman peak of water) is low, recheck the following items. a.
  • Page 129 Maintenance . 129 The above time program shows an example where the minimum FL value was obtained at 380 nm NOTE: and the maximum at 400 nm. For the top Em of the program, set a wavelength that includes the wavelengths where the maximum and minimum FL values were obtained.

  • Page 130: Check And Calibration Of Wavelength Accuracy

    Series 200a Fl User’s Guide 130. Fig. 6-5 Drift Measurement Carry out the drift measurement after the xenon lamp has stabilized (at least one hour after the NOTE: lamp is turned on). Check and Calibration of Wavelength Accuracy 1. Check of Wavelength Accuracy a.
  • Page 131 Maintenance . 131 First adjust the xenon lamp position and then check the wavelength accuracy. An accurate wavelength check cannot be made unless the lamp position is adjusted. If “FAIL” is indicated, wavelength calibration is required. Refer to paragraph (2) below for the calibration.

  • Page 132: Check Of Leak Sensor

    Series 200a Fl User’s Guide 132. Press numeric keys 9 8 and then ENT. ii. The emission wavelength calibration mode screen appears. The present offset value is indicated at Em WL ADJUST. Indicated at the bottom right of the screen is the wavelength check result (deviation of wavelength; indication range of –10 to 10).
  • Page 133 Maintenance . 133 Observe the following precautions before using the leak sensor which CAUTION detects a liquid leak . 1. While a solvent is used, the leak sensor function should be kept active. 2. The leak sensor function of this instrument may not work when liquid leaks in a small amount.

  • Page 134: Periodic Checks

    Series 200a Fl User’s Guide 134. Periodic Checks Check Item Frequency of Check Procedure Check Lighting of lamp Daily Check that “Xe” is indicated on LEDs. Liquid leakage Daily Visually check that the liquid is not leaking from the eluent bottle, tubing and waste bottle Fluorescence Daily...

  • Page 135: Maintenance & Inspection Methods

    Maintenance . 135 Maintenance & Inspection Methods First adjust the xenon lamp position (see chapter 5, Maintenance), then make an energy check of NOTE: the xenon lamp and a wavelength accuracy check using the Hg lamp. Correct measurement results will not be obtained unless the lamp position is adjusted. Energy Check of Xe Lamp Check the energy level of the Xe lamp at a wavelength of 254 nm (see chapter 4, Operation).

  • Page 136: Check And Washing Of Flow Cell Unit

    Series 200a Fl User’s Guide 136. Check and Washing of Flow Cell Unit Ignition of Flammable Chemicals This instrument is not explosion-proof. During unmanned • operation, do not use organic solvents having an ignition WARNING point below 70 °C. Beware of ignition hazard when using flammable chemicals such •...
  • Page 137 Maintenance . 137 Flow cell unit Retaining screw Fig. 6-6 Check of Flow Cell 1. Loosen the retaining screw of the flow cell unit (see the figure above). 2. Pull the flow cell unit straight out. 3. Check the inside of the cell visually to see if it is contaminated with any foreign substance (oil, salt, buffer, etc.).

  • Page 138: Disassembly And Cleaning Of Flow Cell

    Series 200a Fl User’s Guide 138. Fig. 6-7 Injection by Syringe 6. Replace the washing solution with the eluent that is used for separation, and repeat the procedure in step (5). 7. If air bubbles are observed in the flow cell, supply solution into the cell with the pump at a flow rate within 1.0 mL/min and a pressure within 1.0 MPa.
  • Page 139 Maintenance . 139 Fig. 6-9 Flow Cell Unit a. Loosen retaining screws 1 and 2 and detach the shielding plate. b. Detach the cell window. c. Remove the cell clamp. d. Loosen the set screws. e. Take out the flow cell. Fig.

  • Page 140: Leakage From The Flow Cell

    Series 200a Fl User’s Guide 140. NOTE: Make sure to put the flow cell in a paper or plastic container when cleaning it. The use of a glass container may scratch the cell or preclude obtaining the desired performance from the instrument. 3.

  • Page 141: Cautions On Storage Of The Flow Cell

    Maintenance . 141 Fig. 6-12 Inlet and Outlet Tube 1. Supply liquid using the pump, and check that liquid comes out of the OUTLET tube of the flow cell. 2. If liquid does not come out of the tube, it indicates the tube is clogged. Replace the OUTLET tube with a new one.

  • Page 142: Lamp Replacement

    Series 200a Fl User’s Guide 142. Lamp Replacement How to Replace the Xenon Lamp Injury Due to Xenon Lamp Explosion Before removing the xenon lamp for replacement, turn off the xenon lamp (turn off power to the instrument) and then wait for at least one WARNING hour until the xenon lamp becomes sufficiently cool to reduce its internal pressure the normal safety level.
  • Page 143 Maintenance . 143 (c) The Xe lamp setting screen will appear. Press numeric key 0 and then ENT to select lamp OFF. d. The 1st monitor screen now appears, and OFF is indicated at LAMP. e. The xenon lamp is thus turned off. Although the lamp is off, the fan continues rotating so as to cool down the lamp.
  • Page 144 Series 200a Fl User’s Guide 144. Retaining screws Fig. 6-13 Removal of Light Source Cover c. Put on safety goggles for the subsequent steps. d. Detach the connector between lamp house and main body, and loosen the lamp house retaining screw. Connector Flow cell unit Retaining screw...
  • Page 145 Maintenance . 145 Lamp house Grip Fig. 6-15 Removal of Lamp House Fig. 6-16 Lamp House after Removal Removal of old xenon lamp Injury due to Xenon Lamp Explosion (at disposal) The xenon lamp is filled with high-pressure gas (approx. 1 MPa at room temperature, approx.
  • Page 146 Series 200a Fl User’s Guide 146. Injury due to Xenon Lamp Explosion (at mounting) When loosening or tightening the retaining nut for the xenon lamp, be careful not to apply excessive force to its glass bulb part. Never hold and WARNING turn the glass bulb part for loosening/tightening.
  • Page 147 Maintenance . 147 Injury due to Xenon Lamp Explosion (at mounting) Do not touch the quartz glass part of the xenon lamp with bare hands. WARNING If the quartz glass part of the xenon lamp is contaminated with dust or fingerprints, wipe it using a gauze sheet or absorbent cotton cloth slightly moistened with high-quality alcohol.
  • Page 148 Series 200a Fl User’s Guide 148. Fig. 6-18 The Xenon Lamp Fig. 6-19 Orientation of Lamp Bulb Protrusion...
  • Page 149 Maintenance . 149 As shown in Fig. 6-18, put the anode terminal (marked “+”) of the lamp into the metal fixture on the ceramic plate. Then, using the anode nut, mount the lamp so that the protrusion on the bulb is positioned as shown in Fig. 6-19. Refer to Fig. 6- 13 for a view of the lamp when assembled.
  • Page 150 Series 200a Fl User’s Guide 150. Horizontal Focus Vertical Fig. 6-20 Lamp Positional Adjustment a. Coarse adjustment Set the excitation (Ex) wavelength at 550 nm, and make the adjustment while observing the image of the light beam. Take the following steps for the adjustment (see chapter 5, Operation).
  • Page 151 Maintenance . 151 Press the ENT key. v. The 1st monitor screen appears. vi. (vi) In this status, place a paper strip at the Ex beam exit center of the sample compartment as shown in the following figure and observe the condition of the excitation beam.
  • Page 152 Series 200a Fl User’s Guide 152. ii. Press the WL key and the wavelength setting screen appears. Set an excitation (Ex) wavelength. Press numeric keys 3 5 0 and then ENT to set the Ex. iii. Emission (Em) wavelength will not be set here. Leave the present setting as it is.

  • Page 153: Fuse Replacement

    Maintenance . 153 Fuse Replacement Electric Shock due to Contact with Inside of Instrument When replacing the fuse with a new one, be careful not to receive an electric shock. To prevent this, be sure turn off the power switch and WARNING unplug the power cord before proceeding to fuse replacement.
  • Page 154 Series 200a Fl User’s Guide 154.

  • Page 155: Troubleshooting

    Troubleshooting...

  • Page 157: Warnings

    If the instrument becomes faulty, contact the dealer from whom you purchased the instrument or NOTE: the nearest PerkinElmer service engineer. Repairs must be left to the service personnel who have undergone technical training since there are potential hazards.

  • Page 158: Troubleshooting

    158 Series 200a Fl User’s Guide Troubleshooting Symptom Cause Judgment/Check Remedy Self-diagnosis The power cord plug Visual check Plug in the power cord (initialization) is not is not securely securely. performed when the plugged in. POWER switch is turned on. The fuse is blown.
  • Page 159 Troubleshooting Symptom Cause Judgment/Check Remedy The xenon lamp has Check the Xe lamp Replace the xenon lamp reached the end of logbook data. (Refer with a new one. (Refer its useful life. to Section 4.4.4.) to Section 6.2.1.) Excessive drift The warm-up period ____________ Wait until the instrument...

  • Page 160: Error Messages

    160 Series 200a Fl User’s Guide Error Messages Message Description Remedy ROM ERROR ROM is faulty. 1) Press CL key, and 1st PRESS “CL” KEY TO CLEAR monitor screen returns. MESSAGE. 2) Contact our nearest service center. RAM ERROR PRESS “CL” KEY RAM is faulty.
  • Page 161 Troubleshooting Message Description Remedy Xe LAMP ERROR PRESS “CL” Xe lamp fails to come on or 1) Press CL key, and 1st KEY TO CLEAR MESSAGE. go off. Temperature upper monitor screen returns. limiter of Xe lamp power 2) Wait at least 5 minutes source is activated.
  • Page 162 162 Series 200a Fl User’s Guide...

  • Page 163: Appendix

    Appendix Appendix...
  • Page 164 164 Series 200a Fl User’s Guide...

  • Page 165: Description Of Contact Signals

    Appendix DESCRIPTION OF CONTACT SIGNALS Outline The following two types of contact signals are used in the L-2000 series. 1. Contact Signals (START, ERROR, BUSY) Entering the E-Line Connector These are contact signals having input/output functions that are used in common for each unit. These signals are respectively formed from : the START (IN/OUT), ERROR (IN/OUT) and BUSY (IN/OUT) lines.

  • Page 166: Terminology

    166 Series 200a Fl User’s Guide TERMINOLOGY Auto zero [A/Z] Absorbance is electrically adjusted to zero to cancel insignificant parts of data. In common analytical practice, perform auto zero adjustment immediately before or after the start of operation. Auto zero in The external contact signal can be input for auto-zero adjustment.
  • Page 167 Appendix Full scale Used to determine the magnitude of output to such an external device as data processor (chromato-integrator). The confidence function is used to check the status of instrument. Initial screen After power on, the initial screen appears upon completion of initialization. Key lock This protective function prevents an erroneous key input during operation of the instrument.
  • Page 168 168 Series 200a Fl User’s Guide Output range Used to determine the magnitude of output signal to such an external device as data processor. Photomultiplier Used to detect fluorescence emission. Raman scattering In Raman effect, a kind of scattered light appears on the side of wavelength slightly longer than excitation wavelength.
  • Page 169 Appendix Utility Used to set up operating conditions of the instrument. Xenon lamp The xenon lamp is used for fluorescence measurement.
  • Page 170 170 Series 200a Fl User’s Guide...

  • Page 171: Index

    Index Fluorometry advantages ............. 45 descriptions............44 Fuse About This Manual .............10 replacment ............153 Assembling ..............60 Auto Zero adjust ..............76 General Operating Conditions........26 Glossary ..............166 GLP Function Calibration setting ..............84 wavelength ............130 Check flow cell ...............136 lamp logbook............135 leak sensor............132 HG Lamp...
  • Page 172 172 . Series 200a User’s Guide Solvents precautions ............23 safety warnings............22 Spare Parts..............65 Operating Conditions ..........26 Spectrum Measurement..........91 Operation Spectrum Output Factors..........95 adjust auto zero ............76 Static Electricity entering parameters ..........71 cautions ............... 116 GLP function............84 Storage liquid crystal display ..........73 flow cell...............

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