FL-560 - 2025 Operation Manual PDF

MODEL FL 560
Intelligent
Fluorescence Detector
Operators Manual
P/N: KNK-069-270
Safety Considerations
To ensure operation safety, this instrument must be operated correctly and maintained
regularly according to schedule. Carefully read to fully understand all safety precautions in
this manual before operating the instrument. This manual denotes precautions against
actions that can result in hazardous situations or equipment damage by using the signal
words WARNING, CAUTION, and Note.
(1) Safety symbols

Instruction manual symbol. If the product is marked with this symbol, refer
to the instrument manuals to protect the instrument against damage.
WARNING A WARNING indicates an potentially hazardous situation which, if not

avoided, could result in death or serious injury
.
CAUTION A CAUTION indicates a potentially hazardous situation which, if not
avoided, may result in minor or moderate injury. It may also be used to alert
against unsafe practices.
Do not proceed beyond a WARNING or CAUTION notice until you

understand the hazardous conditions and have taken the appropriate steps.
Note A Note provides additional information to aid the operator in obtaining

optimal instrument performance.
Pressurized, hazardous solvents are used in high-performance liquid chromatography.

Always follow the proper laboratory procedures to ensure operator safety. Always wear
goggles, gloves and protective clothing when operating the instrument, especially when
injecting sample and opening valves.
i
(2) Flow Cell Warning
Connecting the outlet of this detector to another detector or tubing with

I.D. of less than 0.5 mm will create an excessive backpressure in the cell
that will result in cell breakage.
(3) Warning Labels

Warning labels are attached at several locations on this instrument. Do not remove,
deface or damage the warning labels. If a warning label peels off the instrument or
becomes illegible, contact your local KONIK distributor (see list on the back cover of this
manual) for a replacement label. Be sure to indicate the part number (P/N) on the label.
(1) Lamp Replacement Warning Label (P/N: 0822-0078A)
WARNING
DISCONNECT POWER TO ALLOW
COOLING TIME BEFORE SERVICING
LAMP.
REPLACE LAMP WITH KONIK
APPROVED LAMP ASSEMBLY ONLY.
USE OF INAPPROPRIATE LAMP
MAY CAUSE BURSTING AND OTHER

PROBLEMS,AND WILL VOID WARRANTY.
TO REPLACE LAMP, FOLLOW
PROCEDURE DESCRIBED IN MANUAL.
The surface temperature of the Xenon lamp can reach more than 200°C when the
lamp is lit. The temperatures of the lamp and the lamp housing do not return to
ambient temperature immediately after the lamp is turned off. Therefore, when
replacing the lamp, take sufficient care to avoid burn injuries.
Refer to the maintenance manual for further details regarding lamp replacement.
Follow the instructions provided therein and perform all operations carefully.
ii
(1) Lamp ReplacemenT Warning Label
Figure 1 Front View of the FL-560
(2) Danger Warning Label (P/N: 0822-0035A)
DANGER
HIG H
V OLTAG E
This label is located on the high-voltage source used to light the Xenon lamp and
operate the photomultiplier tube. A voltage of 200 V or greater is generated during
the operation of the fluorescence detector. Inserting one’s hands or other objects
into this area may result in electric shock. Be sure to turn off the power and
unplug the power cord before performing maintenance in this area.
iii
(3) Xenon Lamp Warning Label (P/N: 0822-0077A)
WARNING
WEAR EYE AND FACE PROTECTION
AND PROTECTIVE GLOVES WHEN
ACCESSING THE XENON LAMP.
THE XENON LAMP CONTAINS HIGH
PRESSURE GAS AND CAN BURST
UNDER MECHANICAL STRESS.
When replacing the Xenon lamp, refer to the operations manual (P/N 0301-
0193A) and follow the instructions carefully.
(4) Attention Warning Label (P/N: 0822-0036A)
ATTENTION
UV RADIATION-EYE PROTECTION REQUIRED
NO USER SERVICEABLE PARTS. DO NOT
REMOVE COVER. DISRUPTION OF CALIBRATION
WILL RESULT IN IMPROPER FUCTIONING OF
UNIT AND WILL VOID WARRANTY.
This label indicates the monochromator and photometer panel. Do not open this
panel under any circumstances! Instrument performance cannot be guaranteed if
this panel is opened by the customer.
(5) Fuse and Ground Warning Label (P/N: 0822-0163A)
iv
(6) Hot Surfaces Warning Label (P/N: 0822-0034A)
The Xenon lamp is located below this label. Do not touch

this area of the instrument because this area becomes hot
while the lamp is lit.
HOT SURFACES
(7) Lamp Replacement Warning Label (P/N: 0822-0079A)
WARNING
REFER TO MAINTENANCE
MANUAL BEFORE SERVICING
LAMP.
Refer to the operations manual for details regarding lamp replacement.
v
(2) Danger warning label
(3) Xenon lamp warning label
(4) Attention warning label
(2) Danger warning label
(6) Hot surfaces warning label
(7) Lamp replacement warning label
Figure 2 Rear View of the FL-560 (Case and Light Source Cooling Fan are Removed)
(5) Fuse and Ground Warning Label
Figure 3 Figure of the side view
vi
(4) Carrying the Instrument
This instrument weighs approximately 18.5 kg. When carrying the instrument, be sure to
hold the instrument as shown in Figure 3.
Figure 4 Grip Locations for Carrying the Instrument
vii
Regulatory Statements
C E Notice
Marking by the symbol indicates compliance of this KONIK system to the EMC
(Electromagnetic Compatibility) and Low Voltage Directives of the European Community.
This symbol indicates that this KONIK system meets the relevant basic safety and health
requirements of the EC Directive based on the following technical standards:
EN55011 ---- ”Limits and Methods of Measurement of Radio Interference

Characteristics of Information Technology Equipment.” ---- Group 1, Class A.
W arning
This is a Class A product. In a domestic environment this product may cause
radio interference, in which case the user may be required to take adequate
measures.
EN50082-1 ---- Electromagnetic compatibility ---- Generic immunity standard Part 1:

Residential, commercial, and light industry.”
IEC61000-4-2 ---- “Electromagnetic compatibility for industrial-process measurement

and control equipment Part 2: Electrostatic discharge requirements.” ---- Severity level 3.

and control equipment Part 3: Radiated electromagnetic field requirements.” Severity
level 2.

and control equipment Part 4: Electrical fast transient/burst requirements.” ---- Severity
level 3.
IEC1010-1: 1990 + Amd.1:1992 + Amd.2: 1995 ---- Safety requirements for electrical
equipment for measurements, control and laboratory use.
IEC61000-3-2: 1995 + Amd.14:2000 ---- “Electromagnetic compatibility: Limits for

harmonic current emissions (equipment input current up to and including 16A per
phase).
A “Declaration of Conformity” in accordance with the above standards has been made
and is on file at KONIK EUROPE srl, Via Confalonieri 25, 22060 CREMELLA (Lc), Italy.
viii
Preface
This instruction manual serves as your guidebook for using this instrument. It is intended
to instruct first-time users on how to properly use the instrument, and to serve as a
reference for experienced users.
Before using the instrument, please read this instruction manual carefully, and make sure that
the contents are fully understood. This manual should be easily accessible to the operator at
all times during instrument operation. When not using the instrument, keep this manual stored
in a safe place. Should this instruction manual be lost, order a replacement from your local
KONIK distributor.
The manuals for this instrument consist of an operations manual and a maintenance manual.
This is the operations manual.
ix
Installation Conditions
To ensure safe operation, the following recommendations should be observed:
(1) Do not operate the instrument under voltage fluctuations exceeding 10% of the
recommended line voltage. Large fluctuations may cause the instrument to fail.
(2) Use a three-pronged electrical outlet with a ground. When only a two-pronged
socket is available, use an adapter and be sure to connect the ground wire of
the adapter.
(3) Operate the instrument under a temperature range of 10 ∼ 30°C.
(4) Operate the instrument under a humidity range of 35 ∼ 85% (RH). If ambient
humidity exceeds 85% (RH), water vapor may deteriorate optical components. If
possible, install the instrument in a location having a humidity of 60% or lower.
(5) Operate the instrument under an atmospheric pressure of 750 ∼ 1060 hPa.
(6) Avoid strong magnetic fields and sources of high-frequency waves. The
instrument may not function properly when near strong magnetic fields or high-
frequency wave sources.
(7) Avoid vibrations caused by vacuum pumps, electric motors, processing

equipment and machine tools.
(8) Avoid dust and corrosive gas. Do not install the instrument in a location where it
may be exposed to dust, especially in locations exposed to outside air or
ventilation outlets that discharge dust particles.
(9) Do not install the instrument in a location where it may be exposed to direct
sunlight.
(10) Do not install the instrument in a location where it may be directly exposed to
the air current from an air conditioner or heater, as such a location may inhibit
stable measurement.
Note: The above conditions do not ensure optimal performance of this instrument.
x
Maintenance
Consult your local KONIK distributor, listed at the back of this manual, regarding
maintenance. In addition, contact your local KONIK distributor representative when
transporting the instrument.
Replacement parts can be ordered according to part number from local KONIK distributor.
When the part number is not known, inform parts name, instrument model name and it’s
serial number.
Notices
(1) KONIK shall not be held liable, either directly or indirectly, for any consequential
damage incurred as a result of the use of this product.
(2) Software prohibitions:

Copying of software or related materials for purposes other than backup is
prohibited.
Transfer or authorization of the use of KONIK software to or by a third party is
prohibited.
Disclosure of confidential information related to KONIK software is prohibited.
Changes or revisions to KONIK software are prohibited.
Use of KONIK software on multiple workstations or terminals, through a network
or through any other means, is strictly prohibited. (This does not apply to
entities with the network license contract)
(3) The content of this manual is subject to change without notice in accordance with
product improvements.
(4) Unauthorized copying of this manual is prohibited.
(5) This manual shall not be used to guarantee or copyright industrial rights or other rights.
(6) Company and product names listed herein are trademarks or registered trademarks of
various companies.
xi
Warranty
This product is warranted for a period of one year from the date of delivery. If any defects
should occur in the product during this period of warranty, KONIK will repair or replace the
defective part(s) or product free of charge.
This warranty does not apply to defects as a result of the following:
(1) USE FOLLOWING IMPROPER OR INADEQUATE INSTALLATION.
(2) IMPROPER OPERATION.
(3) MOVEMENT, MODIFICATION, OR REPAIR BY PERSONS OTHER THAN

AUTHORIZED KONIK PERSONNEL.
(4) USE OF PARTS OTHER THAN THOSE THAT ARE AUTHORIZED BY KONIK.
(5) INORDINATELY RAPID DETERIORATION DUE TO THE USE OF CORROSIVE

SOLVENTS OR SAMPLES.
(6) NATURAL DISASTERS SUCH AS FIRES, WATER DAMAGE, OR

EARTHQUAKES.
In addition, this warranty does not cover:
CONSUMABLE PARTS OR PARTS THAT HAVE A SEPARATE WARRANTY OR

A WARRANTY PERIOD THAT IS DIFFERENT THAN THAT SPECIFIED ABOVE.
The warranty period for all parts and repairs supplied under this warranty expires with the
warranty period of the original product.
KONIK Corporation, 2003
xii
Table of Contents
Safety Considerations..................................................................................i
Regulatory Statements.............................................................................viii
Preface........................................................................................................ix
Installation Conditions................................................................................x
Maintenance................................................................................................xi
Notices........................................................................................................xi
Warranty.....................................................................................................xii
1. Product Description and Specifications...........................................1
1.1 Product Description...........................................................................................1
1.2 Specifications.....................................................................................................1
2. Part Names and Descriptions............................................................4
2.1 Front Panel.........................................................................................................4
2.2 Operation Panel..................................................................................................5
2.3 Flowcell Section.................................................................................................6
2.4 Rear Panel..........................................................................................................7
3. Power-On, Self-Diagnostic Test and Power-Off...............................9
3.1 Power-ON and Self-Diagnostic Test.................................................................9
3.2 Power-OFF........................................................................................................10
4. Operations in Normal Operation Mode...........................................11
4.1 Parameter Settings..........................................................................................11
4.2 Autozeroing......................................................................................................14
4.3 Error and attention Messages During Operation..........................................14
5. [SHIFT] Key Operations...................................................................15
5.1 Description of [SHIFT] Key Operations..........................................................15
5.2 Preamplified Output Display ([SHIFT][1]).......................................................16
5.3 Lamp Off Timer ([SHIFT][2])............................................................................16
5.4 Signal Filter Method Switching ([SHIFT][3])..................................................17
5.5 Automatic Autozero Function ([SHIFT][4])....................................................18
5.6 Integrator Output Zero Point Shift ([SHIFT][5]).............................................20
5.7 Recorder Output Polarity ([SHIFT][6])............................................................22
5.8 Xenon (Xe) Lamp Use Time ([SHIFT][7])........................................................23
5.9 Changing the Spectrum Bandwidth and the Placement of Holmium Glass
([SHIFT][9]).......................................................................................................24
5.10 Changing the Excitation and Emission Wavelength Range
([SHIFT][CLEAR]).............................................................................................24
xiii
5.11 Temperature Display ([SHIFT][MARKER]).....................................................25
6. Operations in Program Mode...........................................................27
6.1 Description.......................................................................................................27
6.2 File Number Setting (File Loading).................................................................27
6.3 Switching Between Normal Operation Mode and Program Mode................28
6.4 Editing Programs.............................................................................................29
6.4.1 Editing initial parameters.............................................................................29
6.4.2 Editing the time program.............................................................................30
6.4.2.1 Time program input..................................................................................31
6.4.2.2 Correcting time programs........................................................................34
6.4.2.3 Deleting individual steps or several steps from a time program...............34
6.4.2.4 Inserting steps into a time program..........................................................35
6.4.3 Program example........................................................................................35
6.5 Time Program Operation.................................................................................38
6.5.1 Running a time program..............................................................................38
6.5.2 Changing parameters while a program is running.......................................38
7. Spectrum Measurement...................................................................39
7.1 Description.......................................................................................................39
7.2 Switching to Wavelength Scan Mode and Measurement Mode Selection..39
7.3 Spectrum Measurement Parameter Settings.................................................42
7.4 Emission and Excitation Spectra Measurement............................................43
7.5 Emission and Excitation Spectra Output.......................................................44
7.6 Emission and Excitation Difference Spectra Output....................................46
7.7 Spectrum Measurement in Time Programs...................................................48
7.7.1 Description..................................................................................................48
7.7.2 Editing time programs.................................................................................49
7.7.3 Time program example................................................................................51
8. Special Accessories.........................................................................52
8.1 Filters................................................................................................................52
8.2 Cell Holder for Square Cells............................................................................53
8.3 Micro-Flowcell..................................................................................................53
8.4 Photomultiplier Tubes.....................................................................................53
9. Error Messages.................................................................................55
9.1 Errors during the self-diagnostic test when power is turned on.................55
9.1.1 ROM, RAM, and DC power errors...............................................................55
9.1.2 Back-up errors.............................................................................................55
9.1.3 Ex/Em drive errors.......................................................................................56
9.1.4 Lamp emission errors..................................................................................56
9.1.5 Slit drive errors............................................................................................57
9.2 Errors during operation...................................................................................57
9.2.1 Dealing with leaks in the flow cell [TROUBLE LEAK IN CELL]...................58
9.2.2 Dealing with wavelength drive problems [TROUBLE EX, EM DRIVE
ERROR]......................................................................................................59
xiv
10. Troubleshooting...............................................................................60
10.1 Noise.................................................................................................................60
10.1.1 Typical noise patterns and possible sources...............................................60
10.1.2 Methods for dealing with noise....................................................................61
10.2 Drift....................................................................................................................63
10.2.1 Typical drift patterns and possible causes...................................................63
10.2.2 Methods for dealing with drift.......................................................................64
11. Maintenance......................................................................................65
11.1 Removing Bubbles from the Cell....................................................................65
11.1.1 Confirming the presence of bubbles through output signal observation......65
11.1.2 Visual confirmation of the presence of bubbles...........................................65
11.1.3 Bubble removal method (1).........................................................................67
11.1.4 Bubble removal method (2).........................................................................68
11.1.5 Bubble prevention method...........................................................................68
11.2 Flow cell maintenance.....................................................................................69
11.2.1 Dealing with flow cell problems...................................................................69
11.2.2 Cleaning the flow cell without disassembly..................................................71
11.2.3 Flow cell disassembly..................................................................................71
11.2.4 Flow cell cleaning........................................................................................73
11.2.5 Flow cell assembly......................................................................................76
11.2.6 Flow cell types.............................................................................................80
11.3 What to do after leaks have occurred............................................................80
11.3.1 Measures for flow cell..................................................................................80
11.3.2 Measures for monochromator.....................................................................81
11.4 Checking and inputting instrument constants..............................................83
11.4.1 Situations requiring that the instrument constants be checked....................83
11.4.2 Location of Instrument Constants Label......................................................83
11.4.3 Input Method for Instrument Constants.......................................................85
11.5 Cleaning the air filter.......................................................................................87
11.5.1 Cleaning the left side-panel air filter............................................................87
11.5.2 Cleaning the rear-panel air filter..................................................................88
11.6 Power Fuse Replacement................................................................................89
12. Replacing Consumable Parts..........................................................90
12.1 Flow cell part replacement..............................................................................90
12.2 Xenon lamp replacement.................................................................................91
12.2.2 Lamp replacement.......................................................................................92
12.2.3 Lamp position adjustment............................................................................97
12.2.4 After replacing the lamp...............................................................................99
12.3 List of consumable parts.................................................................................99
13. Performance Test...........................................................................100
13.1 Checking wavelength accuracy and wavelength calibration.....................100
13.1.1 Definition of wavelength accuracy.............................................................100
13.1.1.1 Wavelength accuracy of the fluorescence (EM) monochromator:......100
13.1.1.2 Wavelength accuracy of the excitation (EX) monochromator:............100
xv
13.1.2 Wavelength accuracy confirmation and calibration for EM monochromator.101
13.1.2.1 Confirmation of fluorescence monochromator wavelength accuracy
using a low-pressure mercury lamp....................................................101
13.1.2.2 Confirmation of fluorescence monochromator wavelength accuracy
using holmium glass...........................................................................104
13.1.2.3 Wavelength calibration for the EM monochromator...........................106
13.1.3 Wavelength accuracy confirmation and calibration for the EX
monochromator.........................................................................................107
13.1.3.1 Confirmation of EX monochromator wavelength accuracy.................107
13.1.3.2 Wavelength calibration for the EX monochromator............................108
13.2 Confirmation of minimum detectable amount.............................................109
13.2.1 Meaning of minimum detectable amount...................................................109
13.2.2 Test method for minimum detectable amount...........................................110
xvi
1. Product Description and Specifications
1.1 Product Description

The FL-560 Intelligent Fluorescence Detector is a high-performance fluorescence
detector designed to provide maximum improvement in detector sensitivity, the most
important requirement of an HPLC detector. The features of the FL-560 are listed
below.
1) Compact, high-efficiency monochromator that incorporates a holographic concave

diffraction grating and a non-spherical mirror.
2) A 150 W Xenon lamp light source.
3) Program functions:
EX wavelength, EM wavelength, gain, attenuation, wavelength scan, and other
parameters can be changed in a time program.
4) Wavelength scan functions:
Excitation spectra and emission spectra can be measured. In addition, difference
spectra can be determined from measured spectra.
5) Maintenance functions:
Maintenance functions include an automatic lamp off function, diagnostic-test function,
functions for checking energy loss, cell leaks, and other items, and the wavelength
calibration function.
1.2 Specifications
Model: FL-560 Intelligent Fluorescence Detector
Optical system:
Monochromator: Holographic concave diffraction grating

(for EX and EM wavelengths)
Light source: 150 W Xenon lamp (mounted horizontally)
Wavelength range: 220 ~ 700 nm (for EX and EM wavelengths)
Spectrum:
Spectrum bandwidth: Excitation side: 18 nm

Emission side: 18 and 40 nm (two-step switching)
Wavelength accuracy: ±2 nm
Wavelength repeatability: ±0.3 nm
Detectors:
Excitation side: Photodiode

Emission side: Photomultiplier
1
Flowcell capacity: 16 µL
Solvent wetted materials: Synthetic quartz, fluoropolimer, and stainless steel
SUS-316
Control system:
Sensitivity: S/N for the Raman peak of water:

standard flowcell(16µL): 350 or greater(FL-560)
450 or greater(FL-650)
micro flowcell(5µL): 150 or greater
where EX = 350 nm and the response is standard
Measurement range: 10 steps in total: 1, 2, 4, 8, 16, 32, 64, 128, 256,

and S
Gain: x1000, x100, x10, and x1
Response: Fast, standard, slow, and digital filter methods
Signal processing: Digital processing by A/D and D/A converters

(having ambient temperature compensation circuits)
Output: Recorder output: 10 mV/FS (polarity change is

possible)
Integrator output: 1 V/FS
Marker output and leak output: 1 circuit each
Input: Marker input, autozero input and Program reset run

input: 1 circuit each
Program functions: Time programmability for EX wavelength, EM

wavelength, gain, attenuation, wavelength scan, etc.
Wavelength scan function: Excitation spectrum and emission spectrum

measurement (manual and time program)
Spectrum storage (10 excitation spectra and 10
emission spectra) and spectrum output (difference
spectrum)
Diagnostic test function: Memory (ROM and RAM), DC power, EX energy

decrease, cell leak, and lamp use time.
Lamp off timer: 99.9-hour maximum
Lamp use time calculation function: Internal timer for recording the total number of
hours that the Xe lamp has been used
Calibration function: EX and EM wavelength calibration
2
Dimensions and weight: 300(W) x 470(D) x 150(H) mm, approx.19 kg
Rated power: AC100 240V±10V%, 50/60Hz, 425 VA
Temperature requirements: +10 ~ +35°C during operation

-30 ~ +60°C during storage
Options:
Photomultiplier: Long wavelength specification (220 ~ 900 nm)

Flowcell: Micro-capacity (5 µL)
Square cell holders: For 10 x 10 mm cells
Secondary light cut filter: For excitation or emission
sides CL kit: Kit for chemical luminescence analysis (optional)
CE cell: Cell for capillary analysis
RS-232C terminal:
* Specifications are subject to change without notice.
3
2. Part Names and Descriptions
2.1 Front Panel

Flow Cell section Operation panel
LCD panel
Power switch Control keys Parameter editing keys
Figure 2.1 Front View
Part name Description

Flowcell section Cassette-type flowcell section
Operation panel Contains an LCD panel (displays operating conditions,
parameters and other items) control keys, and parameter
editing keys
Power switch Used to turn the instrument on and off
4
2.2 Operation Panel
Figure 2.2 Operation Panel
Part name Function

LCD panel Displays operating status, settings, error messages, and other
information.
[PRGM RUN] Used to start and stop the time program. Operating status is
indicated by the lamp to the upper left of the button. The lamp is on
during program execution. This button is also used for spectrum
measurement and output.
[AUTO ZERO] Sets the integrator and recorder outputs to zero.
[MARKER] Adds a marker to the recorder output.
[MONIT] Used to return to or change the monitor screen.
[SHIFT] Used in combination with the numeric keys to set parameters that
are not frequently changed.
[SCAN] Used to switch the wavelength scan mode.
[PRGM] Used to switch between normal operation mode and time program
mode.
[0] ~ [9], [.] Numeric keys have 2 functions. In addition to numeric input, keys [0]
~ [9] are used for special functions, indicated by the alphabetic
characters on these keys. The indicated functions can be accessed
while editing parameters. Refer to Chapter 3 for details regarding
these operations.
[✬ ], [✭ ] Used to change parameter and increase/decrease step number in
the time program mode.
[CLEAR] Used to erase incorrectly entered data and clear errors.
[EDIT/ENTER] Used to edit or confirm parameter changes.
5
2.3 Flowcell Section
Cell body
Cell panel
Flowcell
IN
Lock screws
Figure 2.3 Flowcell Section
Name Function
Flowcell Silica glass flowcell
Cell body Supports the flowcell
Cell panel Holds the flowcell in place
Lock screws Secure the flowcell section to the main unit
<Flowcell attachment method>

Press the cell panel in by hand so that the guide pins on the
main unit go through the holes on the back of the flowcell panel.
Once the flowcell is firmly in place, alternately tighten the lock
screws to secure the cell.
6
2.4 Rear Panel
Light source cooling fan
LC-Net connecter
Ground terminal Power fusee
Line Voltage switch
Figure 2.4 Rear Panel
Name Function
Input and output terminals:
Signal output terminals:
REC± Recorder output terminal (The output level is

G(GND) determined by the ATTEN setting)
INT± Integrator output terminal

G(GND)
MARK+ Marker output terminal (Normally OPEN, the point

OUT- of contact is CLOSED if the [MARK] key is pressed,
or if the MARK IN command is entered.)
LEAK+ Fluid leak warning output terminal (Normally

OUT- OPEN, the point of contact is CLOSED if a fluid
leak occurs in the flowcell.)
Signal input terminals:
MARK IN+ Marker input terminal (A marker input signal is

GND- added to the recorder output if the point of contact
is CLOSED.)
A/Z IN+
GND- Autozero input terminal (The recorder output and
7
integrator output are zeroed if the point of contact
is CLOSED.)
PRGM RST/ST+
GND- Time program input terminal (If the point of contact
is CLOSED, the program starts immediately after
the time program is reset.)
Light source cooling fan Fan for cooling the Xenon lamp
AC input Power cable connection
Power fuses Fuse for power line
F5A(100 ~ 200V),F2.5A(220 ~ 240V)
W arning: For continued protection agains t

risk of fire. R eplace only with fuse
of the specified type and current
ratings.
Ground terminal Terminal for connection to ground

Line voltage selector Switches the line voltage between 115 V and 230 V
115 V: 100 ~ 120 V
230 V: 220 ~ 240 V
LC-Net terminal Used when controlling the detector via a KONIK
system controller
RS-232C terminal (optional) Connection for controlling the detector through the
RS-232C channel.
8
3. Power-On, Self-Diagnostic Test and Power-Off
3.1 Power-ON and Self-Diagnostic Test

Turn the power switch, located at the lower left of the front panel, to the ON position. A
diagnostic test will be performed automatically. The diagnostic test examines the following
areas:
ROM
RAM
DC power (direct current power source)
C-MOS RAM (memory backed-up via battery)
Wavelength drive section
Lamp status (lit or not)
Light quantity
Slit drive section on the emission side
If an error is detected, an ERROR message will be displayed and the self-diagnostic test
will stop. However, if insufficient light is detected, a message will be displayed and the
self-diagnostic test will continue.
Note: W hen an E R R O R message appears, follow the instructions provided in the

maintenance manual.
The screen shown in Figure 3.1 will appear after the diagnostic test has been completed.
Figure 3.1 describes each section of the screen.
Fluorescent
Mode intensity
(Normal operation Response speed
(Standard)
mode)
MODE SIGNAL RESPONSE
NORM 0.0013 STD

300 500 1000 16
EX(nm) EM(nm) GAIN ATTEN
Excitation Emission Gain Attenuation

wavelength wavelength
300 nm 500 nm 1000 16
Figure 3.1 Normal Operation Mode Screen
The wavelength, gain, attenuation and response speed will automatically be set to the
alues that were in effect when the power was turned off.
9
3.2 Power-OFF
Turn the power switch located at the lower left of the front panel to the OFF position. The
wavelength, gain, attenuation, response speed, and time programs saved will be placed in
the C-MOS memory. These values will be restored when the power is turned on again
10
4. Operations in Normal Operation Mode
In normal operation mode the fluorescence intensity is monitored using a fixed excitation
wavelength, emission wavelength, gain, attenuation, and response.
4.1 Parameter Settings

After the power is turned on and the self-diagnostic test is complete, the normal operation
mode screen (Figure 4.1) will appear.
Mode Fluorescent
(Normal operation intensity Response speed
(Standard)
mode)
MODE SIGNAL RESPONSE
NORM 0.0013 STD
300 500 1000 16

EX(nm) EM(nm) GAIN ATTEN
Excitation Emission Gain Attenuation

wavelength wavelength 1000 16
300 nm 500 nm
Figure 4.1 Normal Operation Mode Screen
Key operations for this mode are described below.
(1) Changing the excitation and emission wavelength settings:
The excitation wavelength and the emission wavelength are inputted together.
Input range: 200 ~ 900 nm
11
NORM 0.0013 STD Normal operation mode screen
(Wavelengths will be changed to 230
300 500 1000 16 and 350nm for this example)
[WL/ 1] Wavelength setting
NORM 0.0013 STD Excitation wavelength input screen

300 500 1000 16
Excitation wavelength Press [2][3][0][EDIT/ENTER] to input 230nm

flashes (When the excitation wavelength will not be changed,
simply press [EDIT/ENTER])
NORM 0.0013 STD Emission wavelength input screen

230 500 1000 16
Emission wavelength Press [3][5][0][EDIT/ENTER] to input 350nm

flashes
Figure 4.2 Changing the Excitation and Emission Wavelength Settings
Note 1: If an incorrect value is entered accidentally, press the [C LE AR ] key while the
parameter value is flashing, then enter the correct value. P ress the [E D IT/E NTE R ]
key to enter the setting.
Note 2: The numeric keys are integrated with the parameter keys. W hen a value is
flashing, the keys act as numeric keys.
(2) Changing the gain setting:
Input range: 1,10,100, or 1000

((10)is the new setting.)
300 500 1000(10) 16
[GAIN/3] Press[✭ ] , [✬] [EDIT/ENTER]to advance to the

desired setting
NORM 0.0013 STD Gain input screen
300 500 1000 16
Gain Flashes
Figure 4.3 Changing the Gain Setting
Note: The gain can also be entered using the numeric keys.
12
(3) Changing the attenuation setting:
Input range: 1,2,4,8,16,32,64,128,256, or S

300 500 1000 16(8) (8) is the new setting.)
[ATTEN/ 4] Press [✭ ] [ ✬ ][EDIT/ENTER]to advance to the

desired setting
NORM 0.0013 STD Attenuation input screen

300 500 1000 16
Attenuation Flashes
Figure 4.4 Changing the Attenuation Setting
Note: The attenuation can also be entered using the numeric keys.
Difference between gain and attenuation:
The magnitude of the voltage applied to the fluorescence detector photomultiplier tube is
called gain. Larger gain values mean that a larger voltage is applied and higher sensitivity
is obtained. Gain can be adjusted, allowing a wide dynamic range.
The term attenuation is used to refer to a decrease in the recorder output. Smaller
attenuation values produce smaller decreases in the recorder output, yielding higher
sensitivity. Attenuation adjustment enables the decrease in recorder output to be set
precisely and accurately.
Attenuation does not affect integrator output. Integrator output can be described as
recorder output having attenuation fixed at 1. Recorder output is zero when the
attenuation is set to S.
(4) Changing the response setting:

The input range varies according to the signal filter method being used (See Section 5.4)
Input range: CR FILTER method: FST,STD,SLW
DIGITAL FILTER method: 3s, 5s, 10s, 20s, 40s
NORM 0.0013 STD(SLW) Normal operation mode screen

300 500 1000 16 ((SLW) is the new setting.)
[RSPNS/ 5] Press [ ✭]or [✬ ] to the desired setting

Flashes
NORM 0.0013 STD
Response input screen
300 500 1000 16
Figure 4.5 Changing the Response Setting
13
4.2 Autozeroing
To set the current fluorescent intensity to zero, press the [AUTO ZERO] key. The recorder
and integrator outputs will be set to zero.
4.3 Error and attention Messages During Operation

The error messages that may appear during monitoring are shown in Table 4.2. When an
error message appears, press [CLEAR] to return to the monitor screen. If the problem has
not been solved, the message will reappear after approximately 30 seconds.
Table 4.2 Error Messages
Error message Meaning

LEAK IN CELL There is a leak in the flowcell
WARNING AUTO ZERO Fluorescence signal is too large
LAMP TIMER OFF The LAMP OFF timer has turned the lamp off.
Note: The W AR NING AUTO ZE R O message appears when a bubble is present in the
flowcell or when the gain setting is too high. These situations result in an
excess ive fluorescence signal and proper monitoring is not possible. Use the
procedure des cribed in the maintenance manual to check the flowcell for air. If no
bubble is present in the flowcell, reduce the gain.
14
5. [SHIFT] Key Operations
5.1 Description of [SHIFT] Key Operations

The operations executed using the [SHIFT] key are listed and described in Table 5.1.
Table 5.1 [SHIFT] Key Operations
Key operation Description

[SHIFT][1] Preamplified output display. Used to monitor deterioration of the
lamp or light source mirror and to check for the existence of
bubbles in the cell. The output is also used for re-alignment after
replacing the Xe lamp.
[SHIFT][2] Lamp off timer. The lamp is automatically turned off after the
indicated time has elapsed.
[SHIFT][3] Signal filter method switching (CR FILTER or DIGITAL FILTER).
The signal filter method is normally set at CR FILTER. If a high
noise level makes analysis via the CR FILTER method difficult,
use the DIGITAL FILTER method.
[SHIFT][4] Autozeroing can be performed in the following three modes:
AUTO: When the emission or excitation wavelength, gain,
or spectrum bandwidth is changed, autozero is
performed automatically and the baseline returns
to the zero position. Also, when the contact
closure signal (pulse) is input to the "MARKER IN"
terminals on the rear panel, autozero is performed
atuomatically.
MANUAL: Autozero is performed when the [AUTOZERO] key
is pressed.
HOLD: Autozero is performed and the baseline returns to
the position before autozeroing was performed.
The baseline does not appear to change.
[SHIFT][5] Integrator output zero point shift. The approved input voltage for
the integrator is ordinarily within the range of –10 mV to +1 V.
The dynamic range on the negative side is therefore small
compared to that on the positive side. When the voltage is shifted
beforehand to the positive side using the zero point shift function,
integrator output can be prevented from falling below –10 mV.
[SHIFT][6] Recorder output polarity. The polarity of the recorder output can
be changed using this function. The output polarity change is not
applied to integrator output.
[SHIFT][7] Xenon (Xe) lamp use time. The total time the lamp has been lit
will be displayed on this screen. Use the value here to determine
when to replace the lamp.
[SHIFT][9] Selects the spectrum bandwidth. The spectrum bandwidth can be
switched between 2 levels (18 and 40 nm). Normally the
spectrum bandwidth is set at 18 nm. When wavelength selectivity
15
is not a problem or when the excitation and emission
wavelengths are sufficiently separated, set the spectrum
bandwidth to 40 nm. Analysis at this setting will provide higher
sensitivity.
[SHIFT][CLEAR] Switches the excitation and emission wavelength ranges.
EM>EX+10: Excitation wavelength (EX): 200 ~ 890 nm
Emission wavelength (EM): EX+10 nm ~ 900 nm
NOT RESTRICTED: Excitation wavelength (EX): 0 ~ 900 nm
Emission wavelength (EM): 0 ~ 900 nm
[SHIFT][MARKER] Displays the internal temperature of the instrument.
5.2 Preamplified Output Display ([SHIFT][1])

Preamplified output is used to monitor the deterioration of the lamp or light source mirror
and to check for the presence of bubbles in the cell. The output is also used for re-
alignment after replacing the Xe lamp.
The key operations are shown in Figure 5.1.
NORM 0.0013 STD Monitor screen

300 500 1000 16
[MONIT] [SHIFT] [1]
Excitation light intensity

EX : 2.603 V
Preamplified output screen
Fluorescent intensity EM : 1.865 V
from the flowcell
Figure 5.1 Preamplified Output Screen
5.3 Lamp Off Timer ([SHIFT][2])

The lamp is automatically turned off after the time displayed on the monitor screen has
elapsed.
Note: The lamp off timer is automatically set to O FF when the power is turned on.
16
(1) Setting the lamp off timer:
Input range: 0 (OFF) ~ 99.9 hours (at 0.1-hour increments)
NORM 0.0013 STD

300 500 1000 16 Monitor screen
[MONIT] [SHIFT] [2]

[EDIT/ENTER]
LAMP OFF TIMER LAMP OFF TIMER
xx(yy) HOUR(S) xx HOUR(S)
[EDIT/ENTER]
(yy) is the value after the setting change Flashes
Lamp off timer Time input
Figure 5.2 Setting the Lamp Off Timer
The timer begins to countdown when the time in the lamp off timer screen has been set.
When the designated time elapses, the lamp is turned off and the screen shown in Figure
5.3 appears.
LAMP
TIMER OFF!!
Figure 5.3 Lamp Off Screen
(2) Relighting the lamp:
Turn the power off and then on again.
Note: R elighting the lamp immediately after the lamp has been turned off is s ometimes
difficult. If the lamp will not light, wait for at least one minute, and turn the power
on again.
5.4 Signal Filter Method Switching ([SHIFT][3])

The signal filter method should changed if a high noise level makes analysis difficult. The
method is normally set to CR FILTER. Baseline noise is reduced when the method is
switched to DIGITAL FILTER.
When using the CR FILTER method, FST, STD, and SLW can be selected as the
response.
When using the DIGITAL FILTER method, 3 s, 5 s, 10 s, 20 s, or 40 s can be selected as
the response. However, remember the following when using the DIGITAL FILTER setting.
1) Due to the signal processing principle of the DIGITAL FILTER method, a interval of
approximately 3 times the set value is required before the change will take effect. For
example, when the setting is 5 s, an interval of about 15 seconds is required to
17
process the signal. Therefore, changing parameters immediately before a peak should
be avoided.
2) In addition, the output signals are delayed by approximately 1.5 times the setting
value. For example, when the setting value is 5 s, the peak elution time will be
delayed by about 8 seconds. Therefore, when identifying peaks using retention time,
keep the response setting constant during analysis.
Response settings, signal processing times, and output signal delays are shown in Table
5.2.
Table 5.2 Response Settings, Signal Processing Times, and Output Signal Delays
Signal processing
Response Output signal delay
time
3s 8 s (approx.) 4 s (approx.)
5s 15 s 8s
10 s 30 s 15 s
20 s 60 s 30 s
40 s 120 s 60 s
Key operations are shown in Figure 5.4.
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [3]
[EDIT/ENTER]
SIGNAL FILTER SIGNAL FILTER
CR(DIGITAL) FILTER CR FILTER
[✭ ][✬ ] [EDIT/ENTER]
(DIGITAL) is the new setting Flashes
Signal filter method change screen Screen displayed while changing the signal fitter method
Figure 5.4 Changing the Signal Filter Method
5.5 Automatic Autozero Function ([SHIFT][4])

This function is used to control autozeroing when one of the following parameters is
changed:
Excitation or emission wavelength

Gain
Spectrum bandwidth
When a signal enters the MARK IN terminal on the back panel
18
Input range: AUTO, MANUAL or HOLD
AUTO: Autozero is automatically executed and the baseline of the chromatogram is

returned to zero when a wavelength setting is changed, either in a time
program or manually.
When performing repeated analyses using the autosampler, autozero is
automatically executed with the addition of a marker on the recorder output
when the MARK IN signal is input.
MANUAL: Autozero is not executed when a wavelength is changed or a MARK IN

signal is input.
HOLD: The baseline position will not change when a wavelength is changed or a
MARK IN signal is input. Autozero will be executed, but the baseline position
will return immediately to the position before autozeroing was performed.
Note: This function is not related to the [MAR KE R ] key on the front panel.
(1) Setting the autozero function:

300 500 1000 16
[MONIT] [SHIFT] [4]
[EDIT/ENTER]
AUTO ZERO AUTO ZERO
AUTO(HOLD) AUTO
[✭ ] [✬ ] [EDIT/ENTER]
(HOLD) is the value after the setting change. Flashes
Autozero setting screen Input screen
Figure 5.5 Setting the Autozero Function
Note: As a typical setting, "AUTO " or "HO LD " can be selected for analytical purpose,
and
"MANUAL" for preparative purpose.
19
(2) Autozero examples:
Table 5.3 Autozero examples
Setting Chromatogram change

Wavelength change MARK IN input
AUTO
MANUAL
HOLD
λ1 λ2 MARK IN
AUTO
MANUAL
HOLD
λ1 λ2
MARK IN
5.6 Integrator Output Zero Point Shift ([SHIFT][5])

The approved input voltage for the integrator is within the range of –10 mV to +1 V. The
dynamic range on the negative side is therefore small compared to that on the positive
side. When the voltage is shifted in advance to the positive side using the zero point shift
function, integrator output can be prevented from falling below –10 mV.
Integrator output size during AUTOZERO operations or input of an AUTOZERO signal into
the terminal on the back panel can be selected from 0, 5, 10, 50, or 100 mV.
Note 1: The zero point shift is not applied to recorder output.
20
Note 2: The extent of integrator output nois e is not changed by the zero point shift setting.
However, as the input voltage becomes larger, the signal resolution of the
integrator becomes smaller and the visible baseline noise increases. Therefore,
values should not be set larger than necessary.
(1) Setting the zero point shift:
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [5]
[EDIT/ENTER]
A. Z POSITION A. Z POSITION
(INT OUT) 50(10)mV (INT OUT) 50 mV
[✭][ ✬ ] [EDIT/ENTER]
((10) is the new setting.) Flashes
Zero point shift setting screen Input screen
Figure 5.6 Setting the Zero Point Shift
(2) Application example:
E xample 1: The baseline has a negative drift over a long time period.
Integrator
output
0V
Zero point shift: 0mV
-10 mV
Integrator processing not possible
I ntegrator
output
Zero point shift: +50mV
0V
Figure 5.7 Example of Zero Point Shift Application with Negative Baseline Drift
21
E xample 2: Peaks in the negative direction as with the indirect absorbance method.
Integrator
output
0V Zero point shift: 0mV

-10 mV
Integrator processing not possible
Integrator
output
Zero point shift: +50mV
Waveform processing as negative peaks at the integrator

0V
Figure 5.8 Example of Zero Point Shift Application with Peaks in the Negative Direction
5.7 Recorder Output Polarity ([SHIFT][6])

The polarity of the recorder output can be changed using this function. The output polarity
change is not applied to integrator output.
(1) Changing the recorder output polarity:

300 500 1000 16
[MONIT] [SHIFT] [6]
POLARITY [EDIT/ENTER]
POLARITY
(INT OUT) + (-) (INT OUT) +
[✭] [✬ ] [EDIT/ENTER]
((-) is the new setting.) Flashes
Recorder output polarity change screen Input screen
Figure 5.9 Changing the Recorder Output Polarity
22
Recording as a symmetric reference using another detector to allow easy comparison with
another chromatogram.
FP Polarity: +
RI
FP Polarity: -
RI
Figure 5.10 Recorder Output Polarity Application Example
5.8 Xenon (Xe) Lamp Use Time ([SHIFT][7])

The total time the lamp has been lit will be displayed on this screen. This information can
be used to determine when to replace the lamp.
The key operations are shown in Figure 5.11.
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [7]
LAMP OPERATION
Lamp use time screen
120.5 HOUR
Figure 5.11 Checking Lamp Use Time
23
5.9 Changing the Spectrum Bandwidth and the Placement of
Holmium Glass ([SHIFT][9])
This function is used to change the spectrum bandwidth. Sensitivity or wavelength
selectivity can be improved by changing the spectrum bandwidth. In addition, holmium
glass is placed in the light path for wavelength calibration of the monochromator on the
emission side.
(1) Changing the spectrum bandwidth:
Input range: 18 (standard) or 40
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [9]
[EDIT/ENTER]
BAND WIDTH(nm) BAND WIDTH(nm)
EM : 18(40) EM : 18
[✭ ] [✬ ] [EDIT/ENTER]
((40)is the new setting.) Flashes
Spectrum bandwidth screen Input screen
Figure 5.12 Changing the Spectrum Bandwidth
Note: P ress the [✬ ][✭] keys to display 18 -> 40 -> HO LMIUM while the input screen
shown in Figure 5.12 is displayed. W hen HO LMIUM is selected, holmium glass
will be placed in the light path of the monochromator on the emiss ion side (for
wavelength calibration). This is used when executing wavelength calibration.
When the detected peak is sufficiently separated from other peaks and is a single
component peak having excitation and emission wavelengths that are also sufficiently
separated, fluorescence can be strengthened and sensitivity increased by widening the
spectrum bandwidth. For example, when the excitation and emission wavelengths are
separated by 100 nm or more, sensitivity will be improved if the spectrum bandwidth is set
to 40 nm.
5.10 Changing the Excitation and Emission Wavelength Range

([SHIFT][CLEAR])
This function is used to set the wavelength range for EX and EM.
(1) Changing the wavelength range:
24
The input ranges are shown in Table 5.3.
Table 5.3 Wavelength Ranges
Parameter Wavelength range

EM>EX+10 Excitation wavelength (EX): 200 ~ 890 nm
Emission wavelength (EM): EX+10 nm ~ 900 nm
NOT RESTRICTED Excitation wavelength (EX): 0 ~ 900 nm
Emission wavelength (EM): 0 ~ 900 nm
Note: W hen the wavelength range is set to NO T R E S TR IC TE D , a setting of E X = E M is

pos sible. In this situation, scattered light stronger than the fluores cent light is
incident on the photomultiplier tube of the light detector. To prevent deterioration
of the photomultiplier tube caused by the strong light, preset the gain to a small
value (1 or 10) when setting E X and E M wavelengths that are approximately equal.
Key operations for changing the wavelength range are shown in Figure 5.13.
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [CLEAR]

[EDIT/ENTER]
W.L. RANGE CHECK W.L. RANGE CHECK
NOT RESTRICTED [✭] [✬ ] [EDIT/ENTER] NOT RESTRICTED
The display that appears after making the change is Flashes

not shown here.
Wavelength range screen Input screen
Figure 5.13 Changing the Wavelength Range
5.11 Temperature Display ([SHIFT][MARKER])

This function displays the internal temperature of the instrument. The FL-560
corrects the output signal according to the internal temperature.
25
(1) Temperature display
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [MARKER]
TEMP. 44.9
COF. 1.07959731 ~ 5 digits after the decimal point will fluctuate.
(This degree of fluctuation does not present a
Temperature display screen problem.)
Figure 5.14 Temperature Display
26
6. Operations in Program Mode
6.1 Description
In the program mode, the parameters shown below can be changed according to a time
program. Time programs can contain up to 64 steps, and up to 10 program files can be
stored in the memory of the instrument.
Excitation and emission wavelengths

Gain
Attenuation
Response speed
Autozero
Excitation and emission spectrum measurement
Spectrum bandwidth
In this chapter, the method for setting and performing time programs using the above
parameters, excluding excitation and emission spectrum measurement, will be explained.
The time program for excitation and emission spectrum measurement is unique, and is
therefore described in Section 7.7.
A description of operations in program mode is provided in the flow chart below.
Set file number See Section 6.2
Switch to program mode See Section 6.3
Edit initial parameters See Section 6.4.1
Edit time program See Section 6.4.2

See Section 7.7 for spectrum measurement time
programs
Execute See Section 6.5
6.2 File Number Setting (File Loading)

Up to 10 files (0 ~ 9) can be used. Follow the procedure described below to set the file
number.
(1) Changing file numbers:
27
Input range: 0 ~ 9
NORM 0.0013 STD

Monitor screen
300 500 1000 16
[MONIT] [SHIFT] [PRGM]
[EDIT/ENTER]
PROGRAM FILE NO. PROGRAM FILE NO.
y(x) y
[x][EDIT/ENTER]
((x) is the new setting.) Flashes
File number screen Input screen
Figure 6.1 Changing the File Number
Note 1: If an incorrect value is entered, press the [C LE AR ] key while the parameter value
is flashing, then enter the correct value. P ress the [E D IT/E NTE R ] key to enter the
value.
Note 2: W hen the file number of a file that already contains a time program is designated,
the previous program will be loaded. W hen a time program is edited after setting
the file number, the new file settings are automatically stored. Time programs are
stored in C -MO S R AM and therefore are not erased when the power is turned off.
6.3 Switching Between Normal Operation Mode and Program Mode

Press the [PRGM] key while the normal operation mode screen is displayed. The program
mode monitor screen will appear (Figure 6.2). Press the key again to return to the
previous screen.
NORM 0.0013 STD Normal operation

300 500 1000 16 monitor screen
[PRGM]
PRG5 MON T: 0.0 [MONIT] PRG5 0.0000 STD

300 500 1000 16 300 500 1000 16
Program mode monitor screen
Figure 6.2 Switching Between Normal Operation Mode and Program Mode
28
The program mode monitor screen consists of a time and wavelength screen and a screen
similar to the normal operation mode screen (called the fluorescent intensity screen). Use
the [MONIT] key to alternate between screens. The various screen areas are described in
Figure 6.3.
Current time Fluorescent intensity

Program mode Program mode Response speed
(file No. 5 in this example) (file No. 5 in this example) (standard)

300 500 1000 16 300 500 1000 16
Ex wavelength Gain Ex wavelength Gain

Em wavelength Attenuation Em wavelength Attenuation
Time and wavelength screen Fluorescent intensity screen
Figure 6.3 Description of the Program Mode Monitor Screens
6.4 Editing Programs

6.4.1 Editing initial parameters
Excitation wavelength, emission wavelength, gain, attenuation, and response are set
sequentially as initial parameters.
Note: The excitation wavelength, emission wavelength, gain, attenuation, and response
set in the normal operation mode remain in effect while the instrument is s witched
to program mode.
Press the [ ✭ ] key while the program monitor screen is displayed. The Step 0 screen
(Figure 6.4) will appear. Wavelengths and sensitivity can be changed from this screen
using the procedure described in Figure 6.4.
29

300 500 1000 16 300 500 1000 16
[✭ ] [✬ ] [MONIT]
P5 ST 0 INIT STD Step 0 (Initial parameters screen)

300 500 1000 16 (Note) New values will appear after parameter changes.
[EDIT/ENTER]
P5 ST 0 INIT STD Excitation wavelength input screen
300 500 1000 16
Ex WL flashes Press [3][5][0][EDIT/ENTER] to input 350nm
P5 ST 0 INIT STD Emission wavelength input screen

350 500 1000 16
Em WL flashes Press [5][1][2][EDIT/ENTER] to input 512nm
P5 ST 0 INIT STD Gain input screen

350 512 1000 16
Gain flashes
Press [✭ ] [✬] [EDIT/ENTER] to input 10
P5 ST 0 INIT STD Attenuation input screen

350 512 10 16
Attenuation flashes
Press [✭] [✬ ] [EDIT/ENTER] to input 1
Response flashes
P5 ST 0 INIT STD
Response input screen
350 512 10 1
Press [✭ ] [✬ ] [EDIT/ENTER] to input FST
Figure 6.4 Editing the Initial Parameters
6.4.2 Editing the time program
Note: R efer to S ection 7.7 for details regarding the meas urement of excitation and
emission spectrum as part of a time program.
30
6.4.2.1 Time program input
(1) Time program input example:
As an example, the following time program will be created. First, the excitation wavelength
will be changed to 300 nm and the emission wavelength will be changed to 500 nm
exactly one minute after starting the program under the initial parameters. The gain will
then be changed to one at two minutes.
Press the [ ✭ ] key while the initial parameters screen is displayed (Figure 6.4). The Step 1
screen will appear. Use the procedure described in Figure 6.5 to complete the time
program.
P5 ST 0 INIT STD
Step 0 (Initial parameters screen)
300 500 1000 16
[✭ ] [✬] [✭]
P5 ST 1 T:0.0 Step 1 screen P5 ST 2 T:0.0 Step 2 screen
[EDIT/ENTER] [EDIT/ENTER]
P5 ST 1 T:0.0 Time flashes P5 ST 2 T:0.0 Time flashes

(awaiting input) (awaiting input)
Press [1][EDIT/ENTER] to input 1 minute Press [2][EDIT/ENTER] to input 2 minutes
P5 ST 1 T:1.0 Awaiting function P5 ST 2 T:2.0 Awaiting function

NO.1 EX, EM W.L. input NO.1 EX, EM W.L. input
Press [WL/1][EDIT/ENTER] Press [GAIN/3][EDIT/ENTER]

to call up wavelength to call up the gain
P5 ST 1 T:1.0 Ex WL flashes P5 ST 2 T:2.0 Value flashes

EX: 200 EM: 480 (awaiting input) GAIN 10 (awaiting input)
Press [3][0][0][EDIT/ENTER]
to input 300nm Press [1][EDIT/ENTER] to input 1
P5 ST 1 T:1.0 Em WL flashes P5 ST 2 T:2.0 Step 2 input is

EX: 300 EM: 480 (awaiting input) complete
GAIN 1
Press [5][0][0][EDIT/ENTER]
to input 500nm
Darkened areas in the above figure
P5 ST 1 T:1.0 designate flashing values
Step 1 input is
EX: 300 EM: 500 complete
Figure 6.5 Instructions for Creating the Example Time Program
31
(2) Time program input method:
The input sequence is as follows: time, function, value relative to the function.
1) From the initial conditions screen (Step 0 screen), press the [✭ ] key to advance to
the Step 1 screen.
2) Press [EDIT/ENTER] (time flashes).
3) Enter the time in minutes (minimum increment: 0.1 min).
[xx][EDIT/ENTER]
The function will flash.
4) Input the function. Select a function from Table 6.1.
For example, press [WL/1][EDIT/ENTER]. The function will appear and the value
will flash, awaiting input.
5) Input the desired value.
[xx][EDIT/ENTER]
After Step 1 has been programmed, press the [✭ ] key to advance to the next step.
Repeat steps 1) through 5) to complete the time program. Table 6.1 lists the functions that
can be used in time programs and the corresponding keys used to designate each of the
functions.
Note: Input steps need not be entered in the proper time sequence. W hen the [MO NIT]
key is pressed in order to return to the monitor screen, steps that are out of time
sequence will be rearranged into the proper sequence.
32
Table 6.1 Function Input Keys and Input Ranges
Function Setting key Keys Setting range

Excitation [WL/1] Numeric keys Wavelength range depends on the
wavelength Ex/Em wavelength range setting.
(See Section 5.10.)
Emission
wavelength EM>EX+10:
Excitation wavelength (EX):
200 ~ 890 nm
Emission wavelength (EM):
EX+10 nm ~ 900 nm
NOT RESTRICTED:
Excitation wavelength (EX):
0 ~ 900 nm
Emission wavelength (EM):
0 ~ 900 nm
Gain [GAIN/3] [✬ ][✭ ] x1000, x100, x10, x1
(numeric keys)
Attenuation [ATTEN/4] [✬ ][✭ ] 1, 2, 4, 8, 16, 32, 64, 128, 256, S
(numeric keys)
Response [RSPNS/5] [✬ ][✭ ] Input range varies according to signal
speed filter method. (See Section 5.4)
CR FILTER: FST, STD, SLW

DIGITAL FILTER: 3 s, 5 s, 10 s, 20 s,
40 s
Autozero [A.Z/6]
EM spectrum [EMSP./7] Refer to Section 7.7.
measurement
EX spectrum [EXSP./8] Refer to Section 7.7.
measurement
Spectrum [SLIT/9] [ ✬ ][✭ ] 18, 40 (nm)
bandwidth (numeric keys)
(3) Step advance:
[✭ ]: Advances to the next step

[✬ ]: Returns to the previous step
[SHIFT][✭ ]: Moves to the last step in the time program
[SHIFT][✬ ]: Moves to Step 1
33
6.4.2.2 Correcting time programs
When times and functions are incorrect:
1) Use the [ ✭ ] key to advance to the incorrect step.

2) Enter the correct settings, starting with time.
When changing a setting without changing the time or function:
1) Use the [ ✭ ] key to advance to the incorrect step.

2) Skip correct settings by pressing [EDIT/ENTER] until the desired input screen
appears.
3) Enter the correct setting. For example, when changing only the excitation
wavelength, perform the operations shown in Figure 6.6. However, if settings are
changed starting from time, the function selection screen, shown on the right side of
Figure 6.6, will appear.
PRG ST 3 T: 1.0
W.L 190 nm
[EDIT/ENTER]
P5 ST 3 T:1.0 Time flashes (awaiting input)

EX: 200 EM: 480
Press [1][EDIT/ENTER] to input time
[EDIT/ENTER] P5 ST 3 T:1.0 Function selection screen

(awaiting input)
NO.1 EX, EM W.L.
Press [WL/1][EDIT/ENTER] to call up wavelength
P5 ST 3 T:1.0
Value flashes
EX: 200 EM: 480 (Change the value here.)
Figure 6.6 Correcting time program
6.4.2.3 Deleting individual steps or several steps from a time program
Deleting individual steps:
1) Press the [✭ ] key to advance to the step to be deleted.

2) Set the time in the step to 0 ([EDIT/ENTER][0][EDIT/ENTER]).
3) Press the [MONIT] key to return to the monitor screen. Steps will be rearranged in
sequence and the selected step will be deleted.
34
Deleting all steps following an individual step:
1) Press the [✭ ] key to advance to the first step of the group of steps to be deleted.
2) Press [SHIFT][CLEAR]. The selected step and all subsequent steps will be deleted.
3) Press the [MONIT] key to return to the monitor screen.
6.4.2.4 Inserting steps into a time program
This section describes how to insert a single step into a time program contained in a file.
1) Press [SHIFT][✭ ] to advance to the final step in the program.

2) Press [✭ ] to advance to a blank step.
3) Enter values according to the instructions provided in Section 6.4.2.1.
4) Press the [MONIT] key to return to the monitor screen. Steps will be rearranged in
time sequence.
6.4.3 Program example
Figure 6.7 shows an example of analysis performed while changing wavelengths,

attenuation, and response speed. Detection conditions for the 5 peaks are shown in Table
6.2.
ABS
1 23 5
10 20 30 40 50 Time (min)
Em wavelength: 300nm 370nm 500nm
Attenuation: 32 16 2
Response speed: STD SLOW
Autozero
Figure 6.7 Chromatogram
35
Table 6.2 Peak Detection Conditions
Emission
Peak number/ Response
wavelength Attenuation Note
Condition speed
(nm)
1 300 32 STD
2 370 32 STD
3 500 32 STD (✻ )
4 500 16 SLOW
5 500 2 SLOW
(✻) Immediately following autozeroing
The time program for the above example is shown in Figure 6.8.
36

300 500 1000 16 300 500 1000 16
[✭] [✬]
P5 ST 0 INIT STD Initial parameters screen

Excitation wavelength: 250nm
250 300 10 32 Emission wavelength: 300nm
Gain: 10
Attenuation: 32
[✭] [✬] Response speed: STD
P5 ST 1 T: 8.0 Emission wavelength changed to 370nm

EX: 250 EM: 370 after 8 minutes
[✭] [✬]
P5 ST 2 T: 20.0 Emission wavelength changed to 500nm

EX: 250 EM: 500 after 20 minutes
[✭] [✬]
P5 ST 3 T:27.0
Attenuation changed to 16 after 27 minutes
ATTEN 16
[✭] [ ✬]
P5 ST 4 T:27.0 Response speed changed to SLOW after

RESPONSE SLOW 27 minutes
[✭] [✬]
P5 ST 5 T:41.0
Attenuation changed to 2 after 41 minutes
ATTEN 2
[✭] [✬ ]
P5 ST 6 T:44.0
Autozero executed after 44 minutes
AUTO ZERO
Figure 6.8 Example Time Program
37
6.5 Time Program Operation
6.5.1 Running a time program
Start the program from the program monitor screen.
(1) Starting the time program:
Press the [PRGM RUN] key after the baseline stabilizes. The time program will start. The
[PRGM RUN] key lamp will light, showing that a time program is currently in progress.
(2) Stopping the time program:
Press the [PRGM RUN] key while a program is in progress. The [PRGM RUN] key lamp
be turned off and the time program will stop.
6.5.2 Changing parameters while a program is running
The settings for sensitivity, wavelength, response speed, and lamp off timer can be
changed while a program is running. These changes can be made from the display
screens designated by boxes drawn with double lines in Figure 6.9. The pre-amplified
output can also be displayed.
The settings changes are valid until the program progresses and the next group of settings
are activated.

300 500 1000 16 300 500 1000 16
Time and wavelength monitor screen Fluorescent intensity monitor screen
[SHIFT] [WL/2] [MONIT]
LAMP OFF TIME

2.0 HOUR
Lamp off timer screen

[SHIFT] [WL/1]
EX: 2.603 V
EM: 1.865 V
Pre-amplified output screen
Figure 6.9 Changing Parameters While a Program is Running
38
7. Spectrum Measurement
7.1 Description
The procedures for emission and excitation spectrum measurement and output are
explained in this section.
Measurement of emission or excitation spectra is done manually, after setting the

measurement parameters, at a sample peak while viewing the chromatogram.
Measurement spectra are stored in memory. Up to 10 excitation spectra and 10 emission
spectra (#0 ~ 9) can be stored. Spectra stored in memory can be output to the recorder or
integrator after designating an output range.
The FL-560 can also compare two stored spectra and output a difference spectrum
to the recorder (or integrator). In addition to showing the differences between two samples,
the difference spectrum is used to eliminate fluorescence and scattered light from the
mobile phase.
The FL-560 is also capable of automatically measuring emission and excitation

spectra according to a preset time program. However, output of the measured spectrum
and calculation of a difference spectrum cannot be performed as part of a time program.
Spectrum output and difference spectrum calculations are performed in wavelength scan
mode.
7.2 Switching to Wavelength Scan Mode and Measurement Mode

Selection
(1) Changing modes:
Press the [SCAN] key from the monitor screen during normal operation mode or program
mode. The emission spectrum measurement screen (Figure 7.1) will appear. Press the
[MONIT] key to return to the previous screen.
Normal operation mode Program mode

monitor screen monitor screen
[MONIT] [SCAN]
SPECTRUM MEASURE
NO. 1 EM SPECT
Emission spectrum measurement screen
Fig. 7.1 Switching Between Normal Operation Mode and Wavelength Scan Mode
39
(2) Menu selection:
The [ ✬ ] and [ ✭ ] keys are used to advance through the screens. Figure 7.2 lists the order
in which the screens will be displayed. These seven screens comprise the spectrum
measurement menu. Menus can also be selected by number (See Table 7.1). For
example, press [2] to display the excitation spectrum menu.
Table 7.1 Spectrum Measurement and Output Menus
Number Menu Screen display

1 EM spectrum measurement EM SPECT
2 EX spectrum measurement EX SPECT
3 EM spectrum output EM D.OUT
4 EX spectrum output EX D.OUT
5 EM spectrum difference output DIFFER EM
6 EX spectrum difference output DIFFER EX
0 Measurement parameter settings SCAN PARAM.
40
[MONIT] [SCAN]
Spectrum measurement menus
SPECTRUM MEASURE
EM spectrum measurement
NO. 1 EM SPECT
[✬ ] [✭ ]
SPECTRUM MEASURE
EX spectrum measurement
NO. 2 EX SPECT
[✬] [✭]
SPECTRUM MEASURE EM spectrum output

NO. 3 EM D. OUT
[ ✬] [✭ ]
SPECTRUM MEASURE
EX spectrum output
NO. 4 EX D. OUT
[✬ ] [✭ ]
SPECTRUM MEASURE
EM spectrum difference output
NO. 5 DIFFER EM
[✬] [✭]
SPECTRUM MEASURE
EX spectrum difference output
NO. 6 DIFFER EX
[ ✬] [✭ ]
SPECTRUM MEASURE
Measurement parameter settings
NO. 0 SCAN PARAM
Figure 7.2 Spectrum Mode Screens
41
7.3 Spectrum Measurement Parameter Settings
Both the wavelength scan range and the speed for spectrum measurement can be set.
(1) Parameter input ranges:
The wavelength scan ranges for emission spectra are shown in Table 7.2 and the
wavelength scan ranges for excitation spectra are shown in Table 7.3. Scan speed input
ranges are shown in Table 7.4.
Table 7.2 Scan Ranges for Emission Spectra
Parameter Scan range

WIDE 0 ~ 900 nm
STD EX + 10 ~ 900 nm
NARROW EX + 10 ~ 2xEX (700 nm max.)
Table 7.3 Scan Ranges for Excitation Spectra
Parameter Scan range

WIDE 0 ~ 900 nm
STD 200 ~ EM – 10 nm
NARROW Em/2 ~ EM – 10 nm (220 nm min.)
For example, the EM wavelength scan range is 260 -> 500 nm for the NARROW setting at
EX 250 nm.
Table 7.4 Scan Speeds
Parameter Scan speed

1 100 nm/sec
2 30 nm/sec
(2) Changing the parameter settings:
Figure 7.3 describes how to change the parameter settings.
42
[MONIT] [SCAN]
SPECTRUM MEASURE
Em spectrum measurement menu
NO. 1 EM SPECT
Press [0] (or press [✭] 6 times)
SPECTRUM MEASURE
Parameter settings menu
NO. 0 SCAN PARAM.
[EDIT/ENTER]
SCAN PARAMETERS
Range and speed menu
RNG: STD SPD: 1
[EDIT/ENTER]
SCAN PARAMETERS
Range input menu
RNG: STD SPD: 1
[✬] [EDIT/ENTER]
SCAN PARAMETERS Speed input menu

RNG: WIDE SPD: 1
[✭] [EDIT/ENTER]
SCAN PARAMETERS Input complete

RNG: WIDE SPD: 2 (range and speed menu)
[MONIT]
Figure 7.3 Changing the Parameter Settings
Note: R egardless of the setting, the scan range and speed are automatically set to
NAR R O W and 1, respectively, when a spectrum measurement is performed as
part of a time program.
7.4 Emission and Excitation Spectra Measurement

Spectra can be measured at any time while monitoring the chromatogram.
The emission spectrum measurement procedure is shown in Figure 7.4.
When measuring excitation spectra, instead of the emission spectrum menu shown in
Figure 7.4, display the excitation spectrum menu, set the memory number and emission
wavelength parameters, and then begin measurement.
Note: S can range and scan speed are set using the procedures described in S ection 7.3.
43
[MONIT] [SCAN]
When measuring excitation spectra
SPECTRUM MEASURE Em spectrum
NO. 1 EM SPECT measurement menu SPECTRUM MEASURE Ex spectrum
measurement menu
NO. 2 EX SPECT
[EDIT/ENTER]
EM STECT MEASURE Memory number

input screen
SET MEMORY NO. 0
Press [1][EDIT/ENTER] to
input memory number 1
EX SPECTRUM Em wavelength
EM SPECTRUM Ex wavelength input screen
INPUT EM: 500
INPUT EX: 500 input screen
Press [3][0][0][EDIT/ENTER] to inout 300 nm
EM SPECT. MEASURE Measurement screen

PUSH PRGM RUN
[PRGM RUN] Press [PRGM RUN] to execute measurement
EM SPECT. MEASURE
Screen during measurement
RUNNING
Figure 7.4 Spectrum Measurement Operations
7.5 Emission and Excitation Spectra Output

Emission or excitation spectra can be output to the recorder or integrator.
The magnitude of the output is set using attenuation. Return to the normal operation mode
to set the attenuation.
Wavelength markers will be output for every 100nm from recorder output. No wavelength
markers will be output from integrator output.
The spectrum output speed is approximately 3.3 nm/sec (more precisely: 100 nm/30 sec).
When the chart speed is set to 100 mm/min, a 100 nm spectrum will be contained on 5 cm
of chart.
Figure 7.5 shows an example of the operations required for emission spectrum output.
When outputting an excitation spectrum, display the excitation spectrum menu and use
the same operations described in Figure 7.5.
44
[MONIT] [SCAN]
<Menu selection>
SPECTRUM MEASURE
NO.1 EM SPECT
SPECTRUM MEASURE
Em spectrum output menu
NO.3 EM D. OUT
[EDIT/ENTER]
<Memory number selection>
EM SPECT. NO. 1
Memory number input screen
EX: 350 nm
Press [2] to input memory number 2
EM SPECT. NO. 2
EX: 380 nm
[EDIT/ENTER] <Output range setting>
EM SPECT. EX: 380 Output start wavelength

STRT: 390 END: 650 input screen
Press [4][0][0][EDIT/ENTER] to input 400nm
EM SPECT. EX: 380 Output end wavelength

input screen
STRT: 400 END: 650
EM SPECT. DATAOUT
Output screen
PUSH PRGM RUN
Press [PRGM RUN] to execute the output operation
EM SPECT. EX: 380 Screen during output

(Wavelength and fluorescent intensity
EM: 461 S: 0.013 are shown successively)
Figure 7.5 Spectrum Output Operations
45
7.6 Emission and Excitation Difference Spectra Output
Emission or excitation difference spectra can be output to the recorder or integrator.
The magnitude of the output is set using attenuation. Return to the normal operation mode
to set attenuation.
Wavelength markers will be output for every 100nm from recorder output. No wavelength
markers will be output from integrator output.
The spectrum output speed is approximately 3.3 nm/sec (more precisely: 100 nm/30 sec).
When the chart speed is set to 100 mm/min, a 100 nm spectrum will be contained on 5 cm
of chart.
When calculating an emission difference spectrum, the excitation wavelengths must match
in the source emission spectra. When calculating an excitation difference spectrum, the
emission wavelengths must match in the source excitation spectra. An error message
appears if the wavelengths differ. In addition, the wavelength scan ranges must also
match.
Figure 7.6 shows an example of the operations required for emission difference spectrum
output. When outputting an excitation spectrum, display the excitation difference spectrum
menu and use the operations described in Figure 7.6.
46
[MONIT] [SCAN]
<Menu selection>
SPECTRUM MEASURE
NO.1 EM SPECT
SPECTRUM MEASURE Em difference spectrum output menu

NO.5 DIFFER EM
[EDIT/ENTER] <Memory number selection>
SUB. 3 FROM 2 Memory number input screen

EX: 500 nm
For example, when subtracting the spectrum in Memory 4
from the spectrum in Memory 1, use the following.
SUB. 4 FROM 2
Memory number input screen
EX: 500 nm
Press [1] to input memory 1
SUB. 4 FROM 1
EX: 500 nm
[EDIT/ENTER] <Output range setting>
D. EM EX: 500
Output start wavelength input screen
STRT: 510 END: 650
D. EM EX: 500
Output end wavelength input screen
STRT: 550 END: 650
D. EM DATAOUT
Output screen
PUSH PRGM RUN
Press [PRGM RUN] to execute the output operation
D. EM EX: 500 Screen during output

(Wavelength and fluorescent intensity are
EM: 553 S: -0.7077 shown successively)
Figure 7.6 Emission Difference Spectrum Output Operations
47
7.7 Spectrum Measurement in Time Programs
7.7.1 Description
The elution time of a sample (chromatogram peak position) must be known in order to set
excitation or emission spectrum measurement in a time program.
When creating the time program of spectrum measurement, the time and function
(emission or excitation measurement) are set, then the spectrum measurement
parameters are designated. Parameters include allowable time interval, threshold value for
fluorescent intensity, and memory number. A spectrum will be measured and stored in
memory using the wavelength set at that time during the program when the fluorescent
intensity exceeds the designated threshold within the allowable time interval, the midpoint
of which indicates the designated time.
Spectrum output (including difference spectrum) cannot be performed as part of a time
program. Also, excitation wavelength (while an emission spectrum is measured) and
emission wavelength (while an excitation spectrum is measured) cannot be set as part of
a time program. However, each wavelength is set to the wavelength of the time when a
spectrum measurement (opposite type of spectrum measurement) is started.
A more detailed explanation of spectrum measurement parameters is provided in Table
7.1. Letters in parentheses are displayed on the time program screen.
The wavelength scan range and scan speed cannot be changed (See Table 7.2).
Table 7.1 Spectrum Measurement Parameters
Parameter Function
(Display)
Measurement time Spectrum measurement time, i.e., sample elution time
(T) (chromatogram peak top).
Spectrum type Emission (Fluorescence) spectrum: 7
(7 or 8) Excitation spectrum: 8
Allowable time Allowable time interval with respect to T (spectrum measurement
interval (W) time). The spectrum will be measured within the time interval
from T-W/2 to T+W/2.
Input range: 0.0 ~ 9.9 min
Fluorescent Shows the minimum fluorescent intensity at which spectrum
intensity threshold measurement will be performed, as a proportion of the maximum
value (L) fluorescent intensity (1.0000).
Input range: 0 ~ 99.9%
For example, when 70.0 is set for L, spectrum measurement will
only occur at fluorescent intensities of 0.7000 and above.
Memory number Memory number in which the spectrum will be saved.
(M) Input range: 0 ~ 9
48
Table 7.2 Wavelength Scan Range and Scan Speed
Parameter Setting
Wavelength scan Emission spectrum: EX + 10 ~ 2 x EX nm (700 nm max.)
range Excitation spectrum: Em/2 ~ EM – 10 nm (220 nm min.)
Scan speed 100 nm/sec
Note: Follow the procedures in S ection 7.5 or 7.6 to output a spectrum after a time
program has been performed.
7.7.2 Editing time programs
This section explains the procedure for performing spectrum measurement as part of a
time program.
As an example, the elution time of Peak 2 on the chromatogram in Figure 7.6 is 7.4
minutes and the peak height has a maximum fluorescent intensity of 90%.
100%
70%
Fluorescent
Intensity
0 5 7.4 10
Time (min)
The parameters for measuring the emission spectrum of the peak in Figure 7.7 will be set
as follows in Step 3 of the time program:
Spectrum measurement time (T): 7.4 minutes

Spectrum type: Emission spectrum measurement (7)
Allowable time interval (W): 0.2 min (±0.1 min)
Threshold value for fluorescent intensity (L): Maximum value of 70%
Memory number for storing the spectrum: 5
Note: Letters in parentheses are displayed on the time program screen.
The operations are shown in Figure 7.8.
49
P5 ST 0 INIT STD Step 0
300 500 1000 16 (Initial parameters screen)
[✭] [✬] (Note) Press [✭] 3 times to display the step 3 screen.
P5 ST 3 T:0.0 Step 3 display
[EDIT/ENTER]
P5 ST 3 T:0.0 Time flashes (awaiting input)
Press [7][.][4][EDIT/ENTER] to input 7.4 minutes
P5 ST 3 T:7.4
Function flashes (awaiting input)
NO 1 EX, EM W.L.
Press [7][EDIT/ENTER] to set emission spectrum measurement
P5 ST 3 T:7.4 W (allowable time interval) flashes (awaiting input)

7 W:0.0 L:0.0 M:0
Press [0][.][1] to input 0.1 minute
P5 ST 3 T:7.4
L (threshold intensity) flashes (awaiting input)
7 W:0.1 L:0.0 M:0
Press [7][0][EDIT/ENTER] to input 70%
P5 ST 3 T:7.4
7 W:0.1 L:70.0 M:0
Press [5][EDIT/ENTER] to input 5
P5 ST 3 T:7.4 Step 3 input is complete

7 W:0.1 L:70.0 M:5
Darkened areas in the above figure designate flashing values
Figure 7.8 Setting Emission Spectrum Measurement as Part of a Time Program
Note 1: Input [8][E D IT/E NTE R ] in the function selection screen when the excitation
spectrum is to be measured.
Note 2: S pectrum output cannot be performed as part of a time program.
50
7.7.3 Time program example
Time programs are performed by pressing the [PRGM RUN] key from the program monitor
screen. Refer to Section 6.5 for details.
When the time program described in Figure 7.7 is performed, the emission or excitation
spectrum will be measured when the fluorescent intensity exceeds 0.7000 between 7.3
and 7.5 minutes. The results will be stored in Memory 5. If any of the conditions is not met,
the spectrum will not be measured. The chromatogram during spectrum measurement is
shown in Figure 7.9.
100%
ABU
70%
During spectrum measurement
0 5 7.4 10 (min)
Note: Follow the procedures described in S ection 7.5 or 7.6 to output a spectrum (or
difference spectrum) after a time program has been performed.
51
8. Special Accessories
8.1 Filters
The FL-560 does not usually require the use of a filter. However, when measuring
emission at wavelengths of approximately 2 or 3 times the excitation wavelength, a filter is
required in order to remove the second- and third-order light of the excitation wavelength.
For example, at an excitation wavelength of 250 nm, if the emission wavelength is set at
500 nm, the secondary light of the 250 nm excitation light will be detected along with
fluorescence from the sample. To remove this secondary light, a filter (UV-30, L-1B, L-39,
etc.) that cuts 250 nm light and allows 500 nm light to pass should be attached at the
emission outlet of the flowcell cassette.
Filter applications and characteristics are shown in Table 8.1. The spectral properties of
filters are shown in Figure 8.1 and 8.2.
Table 8.1 Characteristics of Filters for the Fluorescence Detector
Type Application Characteristics

UV-D36C Excitation Band pass filter transmitting light near 360 nm
C-39B Excitation Band pass filter transmitting light near 390 nm
B-460 Excitation Band pass filter transmitting light near 460 nm
UV-30 Emission Sharp cut low pass filter with 50% transmittance near
300 nm
L-1B Emission Sharp cut low pass filter with 50% transmittance near
360 nm
L-39 Emission Sharp cut low pass filter with 50% transmittance near
390 nm
Y-46 Emission Sharp cut low pass filter with 50% transmittance near
460 nm
100 %
B-460
UV-D36C
C-39B
50
B-460
0
200 300 400 500 600 700 nm
Figure 8.1 Transmission Characteristics of Excitation Filters
52
100 %
Y-46
UV-30 L-1B L-39
50
0
200 300 400 500 600 700 nm
Figure 8.2 Transmission Characteristics of Emission Filters
8.2 Cell Holder for Square Cells

A cell holder for 10 mm square cells is available. This cell holder allows the FL-560
to perform intensity measurement using the 10 mm square cells commonly used with the
fluorescent spectrophotometer.
Using these cells, the FL-560 can easily be used as a fluorescent
spectrophotometer. In addition, the difference spectra capabilities of the FL-560
allow the solvent background to be removed from spectra, and difference spectra between
samples can also be output.
8.3 Micro-Flowcell
A micro-flowcell having a volume of approximately 5 µL is available for this fluorescence
detector. This cell can be used to detect narrow peaks using smaller band-broadening
than is normally associated with the standard cell, allowing higher sensitivity analyses to
be performed.
8.4 Photomultiplier Tubes

In addition to the standard tube of the FL-560, a long-wavelength photomultiplier
tube is available.
R928-23 (Long wavelength specification)
While the long wavelength detection limit of the standard photomultiplier tube (R3788-01)
is 700 nm, the limit of the R928-23 long-wavelength photomultiplier tube is 900 nm.
53
The characteristics of each photomultiplier tube are shown in Figure 8.3. These
characteristics are typical and are not guaranteed.
Note: The sensitivity of a fluorescence detector also depends on factors other than the
photomultiplier tube, such as stray light from the monochromator and background
noise. Therefore, replacing the photomultiplier may not always provide the
improvement indicated in the graph.
100
90
Photomultiplier 80
sensitivity R3788-01
(mA/W)
70
60
50
R928-23
40
30
20
10
200 300 400 500 600 700 800 900 nm

Wavelength (nm)
Figure 8.3 Photomultiplier Sensitivity Characteristics
54
9. Error Messages
9.1 Errors during the self-diagnostic test when power is turned on
The FL-560 automatically checks the following items when the power is turned on:
ROM (Memory)
RAM (Memory)
C-MOS RAM (Memory backed up by battery)
DC power (Direct current power supply)
Wavelength drive section
Light source illumination
Light intensity
Slit drive section of the fluorescence monochromator
When there is an abnormality in any of the above areas, with the exception of the light
intensity, an ERROR will be displayed on the LCD and the self-diagnostic test will be
terminated. When the self-diagnostic test detects insufficient light intensity, the ERROR
message will be displayed and the diagnostic test will continue.
9.1.1 ROM, RAM, and DC power errors
When ROM, RAM, DC POWER ERROR appears on the display, turn the power off and
then on again. If the error reoccurs, contact your local KONIK distributor.
Note: After turning off power to the FL-560, wait for at least one minute before
turning the power on again.
9.1.2 Back-up errors
When the contents of the C-MOS RAM have been erased, BACK UP ERROR will appear
on the display. When this happens, the instrument constants and various parameter
settings will be erased and the default settings will be restored. If a BACK UP ERROR
occurs, the procedures listed below should be followed.
(1) While holding down the [SHIFT] key, press the [✭ ] key to continue the self-
diagnostic test.
(2) When the self-diagnostic test has been completed and the monitor screen
appears, refer to Section 11.4 and input the instrument constants. Input the
wavelength, sensitivity etc, and other values inputted after [SHIFT] + [n]
operation. This should rectify the error, and the instrument should operate
normally until the power is turned off again.
(3) If the error reoccurs the next time the power is turned on, the battery should
be replaced. Contact your local KONIK distributor for information on this
procedure.
55
Turn power on
BACK UP ERROR
[SHIFT] [✭]
Continue diagnostics test
NORM 0.0000 STD

Monitor screen
400 500 1000 128
Input instrument constants See Section 11.4
Check settings and make In addition to the wavelength and sensitivity est,
revisions input as necessary be sure to check the settings displayed using
the [SHIFT]+[N] operation.
(refer to the operations manual).
Figure 9.1 Dealing with Back-up Errors
9.1.3 Ex/Em drive errors
An EX, EM DRIVE ERROR is displayed when a problem occurs in the drive that rotates
the diffraction grating, turn the power off and then on again. If the error reoccurs, contact
your local KONIK distributor.
9.1.4 Lamp emission errors
LAMP EMISSION POOR will appear on the display. When the excitation light is
insufficient, The following are possible causes of insufficient light:
Lamp deterioration
Light source mirror deterioration
Improper adjustment of lamp or light source mirror
Deterioration of optical elements in the monochromator
Note: The LAMP E MIS S IO N P O O R error does not indicate an instrument failure, only a
reduction in the excitation light inteusity therefore, the instrument may continue to
be used. However, a significant increas e in noise may result in an instrument
failure. In which cas e, the lamp emission must be improved.
56
Note: A LAMP E MIS S IO N P O O R error will appear on the display when the output from
the excitation pre-amplifier (E X ) falls below 0.40 V for an excitation wavelength of
400 nm.
A LAMP EMISSION OFF! error will appear on the display when the lamp will not light.
When this error occurs, turn the power off, wait for approximately one minute, and then
turn the power on again. If the problem continues to occur despite performing this
procedure several times, lamp deterioration or a damaged lamp power source are likely to
be the source of the problem. If a spark-like sound can be heard when the power is turned
on, the problem is caused by lamp deterioration and the lamp should be replaced. If no
spark-like sound can be heard when the power is turned on, the problem is being caused
by a damaged lamp power source. Contact your local KONIK distributor for information on
procedures for dealing with this problem.
9.1.5 Slit drive errors
The SLIT DRIVE ERROR message appears on the display when a problem occurs in the
Em slit drive, turn the power off and then on again. If the error reoccurs, contact your local
KONIK distributor.
9.2 Errors during operation

Table 9.1 lists the problems that can be detected by the FL-560 during operation.
When a problem is detected, a message will appear on the LCD. Press the [CLEAR] key
to return to the monitor screen. If the problem reoccurs, the error message will reappear
after approximately ten seconds.
Table 9.1 Error Messages During Operation
Error Messa ge Meaning

TROUBLE LEAK IN CELL The flow cell has a leak
TROUBLE EX, EM DRIVE ERROR A wavelength drive problem exists
When the setting time of the lamp-off timer has elapsed, the LAMP TIMER OFF message
will appear and the lamp will turn off. This is not an error message.
To relight the lamp after it has been turned off by the timer, turn the power off and then on
again.
Note: After turning the power off, wait for at least one minute before turning the power
on again.
57
9.2.1 Dealing with leaks in the flow cell [TROUBLE LEAK IN CELL]
When a TROUBLE LEAK IN CELL error occurs, stop the solvent flow, remove the flow
cell, and inspect the inside of the instrument. Refer to Section 11.3 for information on
procedures for dealing with solvent leaks.
In order to minimize the potential for leak-induced damage to the FL-560, the FL-
560 should be wired to the pump as shown in Figure 9.2. This wiring
arrangement stops the pump immediately upon detection of the TROUBLE LEAK IN CELL
error.
Note: This wiring arrangement is not required when the pump and the FL-560 are
controlled by the HS S system controller.
FL-560 PUMP
Figure 9.2 Leak Output Wiring
58
9.2.2 Dealing with wavelength drive problems [TROUBLE EX, EM DRIVE ERROR]
Refer to Section 9.1.3 for information on procedures for dealing with wavelength drive
problems.
59
10. Troubleshooting
This chapter provides an explanation of noise and drift, which are typical chromatogram
problems, and possible methods for dealing with these problems. Although noise and drift
may arise for a variety of reasons, many of which are not associated with the detector, this
chapter focuses primarily on noise and drift that occur due to problems in the detector.
10.1 Noise
10.1.1 Typical noise patterns and possible sources
Table 10.1 is a list of typical noise patterns and possible sources associated with each
pattern.
Table 10.1 Typical Noise Patterns and Possible Sources
Noise pattern Probable cause

Always appears in the Bubble in cell
direction of greater
fluorescence
Bubble in cell
Suspension in cell
Pumping stopped
Lamp energy reduction

Cell contamination
Lamp noise
Under
normal conditions
Electronic noise
Lamp not lit
60
10.1.2 Methods for dealing with noise
Table 10.2 is a list of methods for dealing with noise from various sources.
Table 10.2 Methods for Dealing with Noise
Source Confirmation Method What to Do

(1) Bubble in the Lightly block the tubing on the outlet side Install a back-
cell of the flow cell. This causes pressure pressure coil
variation within the cell, and if the baseline (See Section 11.1)
varies greatly, a bubble is present in the Remove the air
cell. from the solvent
Monitor the baseline variation when fluid Use a degasser
is being pumped and when the pump is
stopped. Bubbles cause abnormally large
baseline variation when the flow rate
changes.
(2) Suspension in Use a magnifying glass to carefully Disassemble and
the cell examine the cell. clean the cell
Check baseline variation when fluid is (See Section 11.2)
being pumped and when the pump is
stopped. Suspension causes abnormally
large baseline variation when the flow rate
changes.
(3) Fluid is leaking Remove the flow cell from the instrument. Disassemble and
from the cell Examine both the flow cell and the inside clean the cell
of the instrument. (See Section 11.2)
(See Section 11.3
regarding
instrument care)
(4) Energy Set the excitation wavelength to 400 nm Replace the lamp or
reduction and check the output from the excitation mirror
pre-amplifier by pressing [SHIFT] [1]. (See Section 12.2)
If the Ex is ≤ 0.4 V, the lamp or mirror has
deteriorated.
(5) Lamp noise Set the excitation wavelength to 400 nm Replace the lamp
and check the output from the excitation (See Section 12.2)
pre-amplifier by pressing [SHIFT][1]. If the
Ex variation is ≥ 5%, the lamp has
deteriorated.
(6) Cell Measure the fluorescent spectrum for the Remake the mobile
contamination solvent alone. If the fluorescent intensity is phase
or Solvent abnormally high, either the cell or the Use a high-grade
contamination solvent may be contaminated. solvent
(Increase in Check the
scattered light wavelength
or fluorescence) Disassemble and
clean the cell
(See Section 11.2)
61
(7) Electronic noise Ground the
instrument
(8) Lamp is not lit Set the excitation wavelength to 400 nm Replace the lamp
and check the pre-amplified output for (See Section 12.2)
excitation by pressing [SHIFT][1]. If Ex = If the lamp power
0.000 ~ 0.005 V, the lamp is not lit. source appears to
If a spark-like sound can be heard when be damaged,
the power is turned on, the problem is contact your local
caused by lamp deterioration; otherwise, KONIK distributor.
the problem is caused by a damaged
lamp power source.
62
10.2 Drift
10.2.1 Typical drift patterns and possible causes
Table 10.3 is a list of typical drift patterns and possible causes associated with each
pattern.
Table 10.3 Typical Drift Patterns and Possible Causes
Drift Pattern Possible Cause
Fluorescent impurities in the solvent

are flowing through the cell
Increase in scattered light due to cell
Irregular drift contamination
Note: In addition to detector-related
problems , column or injector
contamination may cause drift.
Increase in bubble size
Drift toward greater

fluorescence
Decrease in bubble size
Drift toward lesser Note: In addition to detector-related

fluorescence problems, column contamination or
ins ufficient replacement of s olvent may
cause drift.
The detector or column may be

exposed to direct air flow from an air
Periodic drift conditioning duct.
Bubbles may be gathering in the cell
and then passing through it.
63
10.2.2 Methods for dealing with drift
Table 10.4 is a list of methods for dealing with drift from various sources.
Table 10.4 Methods for Dealing with Drift
Cause Confirmation Method and What to Do

(1) Bubble in the cell See (1) in Section 10.1.2
(2) Fluid is leaking from See (3) in Section 10.1.2
the cell
(3) Increase in See (6) in Section 10.1.2
fluorescence or
scattered light
(4) Cell contamination See (6) in Section 10.1.2
64
11. Maintenance
11.1 Removing Bubbles from the Cell

The presence of bubbles in the cell can produce noise and drift. In organic solvents, the
solubility of air is high so bubbles that remain in the cell disappear relatively quickly.
However, in water, the solubility of air is low and bubbles tend to remain in the cell when
water is used as a solvent. Refer to in Section 10.1.1 and 10.2.1 for a description of
baseline phenomena that occur when bubbles are present in the cell.
11.1.1 Confirming the presence of bubbles through output signal observation
This section describes how to confirm the presence of bubbles using the fluorescence
output signal. When bubbles remain within the cell, the excitation light reflects at the
boundary between the bubble and the solvent. The light is then emitted to the
fluorescence monochromator, appearing as an increase in the apparent fluorescence
signal. When pressure within the cell is varied, the bubble moves, causing the extent of
the reflection to vary. This appears as a variation in the fluorescence signal. Since the
actual fluorescence is normally influenced little by pressure variations within the cell,
bubbles are confirmed to be in the cell when the fluorescence signal varies greatly due to
changes in pressure.
The method for varying the pressure within the cell is described below.
CAUTION: The flow cell can withstand a pressure of approximately 1.0 MPa. If the
outlet of the flow cell is obstructed to a high degree, the cell may be
destroyed.
(1) Attach a stainless steel tube to the outlet of the flow cell and cover the stainless
steel tube with a Teflon tube (o.d.: 2 mm, i.d.: 1.4 mm)
(2) Repeatedly cover and uncover the end of the Teflon tube using your fingertip. If
the baseline varies greatly, a bubble is probably present in the cell.
W AR NING: The Teflon tube may come off of the stainless steel tube if too much
pressure is applied. Be careful not to cover the end of the Teflon tube
for a long time, since solvent may splatter when these tubes come
apart.
Note: In addition to the above method for varying the pressure within the cell, the flow
cell outlet may also be covered and uncovered directly with your fingertip. D o not
use this method when using dangerous solvents.
11.1.2 Visual confirmation of the presence of bubbles
The method used to visually confirm the presence of bubbles in the cell is explained in this
section.
65
(1) Refer to Figs. 11.1 remove the flow cell from the instrument. If measures have
been taken to prevent solvent leakage, the flow cell may be removed while the
solvent is still being pumped.
Driver
Hold here and pull out.

The unit is easier to pull out
when the tubing is attached
Expanded view of the cell portion
Figure 11.1 Flow Cell Removal and Attachment
66
(2) Inspect the inside of the cell under good lighting conditions (See Figure 11.2)
Cell
Out
In
Locations where bubbles easily remain

Flow route
Figure 11.2 Visual Check for Bubbles in the Cell
11.1.3 Bubble removal method (1)
Remove bubbles using the procedure described in Figure 11.3.
Increase the pump flow rate
If bubbles remain,
Connect the outlet port of the pump

directly to the IN side of the cell and
pump at the maximum flow rate.
If bubbles remain,
Use a syringe to inject methanol and

remove the bubbles. After removal,
return to the original solvent.
Note: W hen buffers or salt solvents are being used, first replace the solvent with water
and then replace the water with methanol. B e sure to replace the methanol with
water before restoring the original solvent.
Figure 11.3 Bubble Removal Method (1)
67
11.1.4 Bubble removal method (2)
Bubbles may be compressed and pass through the cell if pressure is applied within the
cell (See Figure 11.4).
(1) Attach a stainless steel tube to the outlet of the flow cell and cover the stainless
steel tube with a Teflon tube (o.d.: 2 mm, i.d.: 1.4 mm).
(2) Repeatedly cover and uncover the end of the Teflon tube using your fingertip (the
tube may also be bent and released). If the baseline moves in the negative
direction (toward smaller fluorescence), the bubbles in the cell are decreasing in
size.
W AR NING: The Teflon tube may come off of the stainless steel tube if too much
pressure is applied. Be careful not to cover the end of the Teflon tube
for a long time, since solvent may splatter when these tubes come
apart.
Note: In addition to the above method, pressure can be varied by directly stopping and
releasing the stainless steel tube. However, when using dangerous solvents, do
not interrupt solvent flow using your fingers.
Teflon tube
Figure 11.4 Bubble Removal Method (2)
11.1.5 Bubble prevention method
The most effective method for preventing the generation of bubbles is the use of a
degasser. In addition to preventing bubble generation, the degasser is also effective in
stabilizing the pump operation.
If a degasser is not available, attach a back pressure coil to the cell outlet in order to
prevent the generation of bubbles. Attach a stainless steel tube to the outlet of the flow
cell, to act as a back pressure coil, so that the back pressure will be approximately 0.1
MPa under pumping conditions. Although the back pressure coil can suppress the
generation of bubbles, it cannot remove bubbles that initially remain in the cell. Attach the
back pressure coil after referring to Sections 11.1.3 and 11.1.4 and removing the bubbles.
68
CAUTION: (1) Do not pump at a high flow rate when the back pressure coil is
attached. The cell may be damaged if the flow rate is too high.
(2) If pumping is performed while the back pressure coil is clogged,

the cell may be damaged. Be sure to check for clogs in the back
pressure coil before starting the pump. In particular, take special
care when buffers are being used, because salts precipitate and
can easily clog the tube.
11.2 Flow cell maintenance

11.2.1 Dealing with flow cell problems
(1) Fluid leaks

(2) Suspension
(3) Contamination of the cell wall
These problems may be detected while searching for the causes of noise and drift on a
chromatogram. Use the flow chart below to correct the problem.
(1) Fluid leaks (LEAK IN CELL appears on the LCD display and/or fluid has leaked from
the bottom of the FL-560.)
Turn off the power to the FL-560, and

See Fig. 11.2
then remove and inspect the flow cell.
Is it clear that the cell is broken? Yes Go to Sections 11.2.3,

11.2.4, and 11.2.5
No
Inject methanol into the IN side of the flow

cell using a syringe or other device and
answer the following questions: Yes
1. Is a fitting leaking? Tighten the fitting.
2. Is the cell leaking? Yes
Go to Sections 11.2.3,
11.2.4, and 11.2.5
No
69
Return the flow cell to the FL-560 and
turn the power on. Yes
Does the LEAK IN CELL message appear Go to Section 11.3
again and continue to re-appear after the
[CLEAR] key is pressed?
Figure 11.5 Dealing with Fluid Leaks
(2) Suspension
Remove and inspect the flow cell.

Is there suspension?
Yes
Alternately and repeatedly inject air and then

methanol using a syringe. Yes
End
Was the suspension removed?
No
Go to Sections 11.2.2, 11.2.3, 11.2.4, and 11.2.5
Figure 11.6 Dealing with Suspension

(3) Contamination of the cell wall
Remove the flow cell and inspect the cell wall

for contamination. Is contamination present?
Yes
Follow the instructions given in Section 11.2.2 Yes

Was the contamination of the cell wall End
removed?
No
Go to Sections 11.2.3, 11.2.4, and 11.2.5
Figure 11.7 Dealing with Contamination of the Cell Wall
70
11.2.2 Cleaning the flow cell without disassembly
Contamination of the cell wall can sometimes be removed by flowing organic solvents or
strong nitric acid through the cell. The procedure is described below. When sufficient
results cannot be obtained using this method, disassemble and clean the flow cell.
CAUTION: When replacing the solvent inside the flow cell with distilled water, first
confirm that the solvent being used is intermiscible with distilled water.
If an immiscible solvent is being used, first change to a solvent that is
intermiscible with both water and the original solvent, and then replace
the solvent with water. When returning to the original solvent, reverse
the procedure.
Replace the solvent in the flow cell with

distilled water.
Flow concentrated nitric acid through the

cell (7 N, 1.0 mL./min, 20 min).
Flow distilled water through the cell to

sufficiently clean the cell
(1.0 ~ 2.0 mL/min, 20 min)
Flow acetone through the cell

(1.0 ~ 2.0 mL/min, 20 min).
Flow distilled water through the cell to

sufficiently clean the cell
(1.0 ~ 2.0 mL/min, 20 min)
Figure 11.8 Method for Washing the Flow Cell
11.2.3 Flow cell disassembly
Disassemble the flow cell and wash or replace parts when:

the cell is broken (fluid leaks) and requires replacement
suspension is present in the cell
the cell wall is contaminated
the gasket requires replacement
71
(1) (11)
1 hole (2)
(3) (16)
Gasket on
the OUT side
(4)
(10)
(5) (12)
4 holes
(6)
(5)
(18)
(19)
Gasket on
(8) (17)
the IN side
(15)
(14)
(20) (21)
(20)
(13) (7) (9)
(1) (OUT) tubing (9) (IN) tubing (16) Unions

(2) Cell retaining screw (10) Cell body (17) Collars
(3) Seat (11) Cell panel (18) Pin
(4) (OUT) cell holder (12) Cell panel retaining (19) Sunken-head screw
(5) Gaskets screw (20) Screw
(6) Cell (13) Mask retaining screw (21) Filter holder
(7) (IN) cell holder (14) Pin
(8) Cell mask (15) Pin hole
Figure 11.9 Flow Cell Diagram
Refer to Figure 11.9 and use the following procedure to disassemble the flow cell.
(1) Remove the cell panel retaining screw (12).

(2) Use the standard accessory wrench to loosen and remove the tubing from the unions
(16).(See Fig 11.10)
72
Loosen and remove Loosen and remove this cell panel
these tubings retaining screw
Figure 11.10 Tubing Removal
(3) Remove the cell panel (11) (with the unions (16) attached) from the cell body (10).
(4) Loosen the cell mask retaining screw (13) and remove the cell mask (8).
(5) Loosen the cell retaining screw (2) while noting the tightness of the screw. When
reassembling the flow cell, tighten the cell retaining screw to this tightness (See 7 of
Section 11.2.5).
(6) Remove the cell retaining screw (2) and pull the (OUT) cell holder (4) out of the cell
body (10). The (OUT) cell holder (4) can be removed with the (OUT) tubing (1) still
attached.
(7) Remove the gaskets (5) from the (OUT) cell holder (4) and the (IN) cell holder (7).
Note 1: W hen the cell is damaged or fluid is leaking from the cell, the (O UT) tubing (1),
the (IN) tubing (9), the (O UT) cell holder (4), the (IN) cell holder (7), or the unions
(16) may be clogged. Flow distilled water through each of thes e parts to confirm
that no clogs are present.
Note 2: D is assemble the flow cell and remove the cell before replacing either the (O UT)
tubing (1) or the (IN) tubing (9).
11.2.4 Flow cell cleaning
Clean the cell when:

contamination could not be removed using the washing method described in
Section 11.2.2
oil from the user’s hands is on the cell.
salt or other materials have precipitated onto the cell, the gasket, or other parts.
CAUTION: (1) The cell is made of quartz glass. Like ordinary glass, quartz is
easily broken. Therefore, take particular care not to chip the
corner sections. Chipped corners may result in increased light
scattering and reduced sensitivity. The cell should be handled
using tweezers that have been covered with Teflon tubing (See
Figure 11.11).
73
(2) Do not reuse gauze or laboratory tissues as this may cause cell
contamination.
(3) The surface of the cell wall has an aluminum coating and a
protective film. If the aluminum coating is peeled or scratched
only slightly, the optical capabilities of the cell will not be
influenced. However, if the surface is rubbed off by the sharp ends
of tweezers or by other objects, the protective film may peel off.
Peeling or scratches in the protective film may result in corrosion
of the aluminum coating. Be careful when handling the cell.
(4) If the cell is soaked in organic solvent for more than three minutes,
the aluminum coating of the cell wall may peel. When soaking the
cell in organic solvents, soak only for one minute or less.
Easily chipped areas
Figure 11.11 Areas of the Cell that Are Easily Chipped
Wash the cell according to the following procedures:
Soak the cell in warm water and perform Warm water at approximately 40°C is
ultrasonic cleaning for approximately ten appropriate.
minutes. Cleaning is more effective when low residue
soap suitable for washing glass laboratory
equipment is used.
Flush out the soap using distilled water. Be sure to flush sufficiently, because
soap may generate fluorescence.
Soak the cell in methanol for several

seconds and then remove the cell.
74
Dry the outer wall surface of the cell using
gauze or laboratory tissue.
Figure 11.12 Cleaning the Cell
75
11.2.5 Flow cell assembly
Assemble the flow cell according to the following procedures.
CAUTION: (1) When the flow cell has been disassembled in order to service a
broken cell or stop a fluid leak, be sure to check for clogs in the
(OUT) tubing (1), (IN) tubing (9), (OUT) cell holder (4), (IN) cell
holder (7), and unions (16) before re-assembly. The cell may be
broken or leaks may reoccur if the flow cell is assembled using a
clogged part.
(2) Do not reuse gauze or laboratory tissues as this may cause cell
contamination.
(3) Be careful of alignment when inserting the cell (See Figure 11.16).
If the cell is inserted incorrectly no fluorescence signal will be
obtained. Cells that have no aluminum coating (optional) may be
inserted in either direction.
(1) Place new gaskets (5) over the (OUT) cell holder (4) and the (IN) cell holder (7).
4 holes 1 hole
OUT gasket
5mm
5mm
1 hole
OUT
5mm
Al coated cell LD gasket Gasket

Replacement kit (LD 16uL) P/N:6715-H311B
4 holes 4 holes 1 hole

IN
5mm
5mm 5mm
IN gasket LD gasket Gasket

Microcell
Replacement kit (LD 5uL) P/N:6715-H316B
Figure 11.13 Expanded View of Gaskets (5)
(2) Insert the (OUT) cell holder (4) and the seat (3) into the cell body (10), making sure
that the groove of the (OUT) cell holder (4) and the pin of the cell body (10) are
aligned (See Figure 11.14).
76
Cell holder (OUT) (4) Groove Pin Cell body (10)
Figure 11.14 Insertion of (OUT) cell holder
(3) Insert the cell retaining bolt (2) into the cell body (10) and tighten until the cell can be
inserted between the (OUT) cell holder (4) and the (IN) cell holder (7) (See Figure
11.15).
Cell body (10)

Cell retaining screw (2)
Tighten until the cell can be

inserted between the cell
body and the cell retaining
screw
Figure 11.15 Tightening the Cell Retaining Screw (2)
(4) Insert the cell between the (OUT) cell holder (4) and the (IN) cell holder (7). In order to
prevent oil from your fingers from coming into contact with the outer wall of the cell (6),
use laboratory tissues or tweezers covered with Teflon tubing. Insert the cell (6) in the
correct direction (See Figure 11.16).
Aluminum coated surface

Cell panel (11)
Expanded figure of cell
Insert in this direction
Excitation light
Figure 11.16 Cell (6) Insertion Direction
77
(5) Tighten the cell retaining screw by hand (2) while making sure that the cell (6) does
not come out from between the (OUT) cell holder (4) and the (IN) cell holder (7).
When the cell (6) has been lightly secured, stop tightening the cell retaining screw (2).
(6) Position the cell (6) correctly (See Figure 11.17).
These surfaces should

be aligned
Correct Incorrect
Figure 11.17 Cell (6) Positioning
(7) Tighten the cell retaining screw (2) using a wrench. After sufficient hand tightening,
the proper tightness will be achieved with a 30 ~ 45° turn of the cell retaining screw
(2) (Refer to Step 5 of Section 11.2.3. The specified torque is 1.0 MPa).
(8) Wait approximately 30 minutes and then tighten the cell retaining screw (2). (This is
necessary because the gaskets (5) will compress.)
(9) Attach the cell panel (11) to the cell body (10) (See Figure 11.18).
Sell panel retaining screw (12) Pin (14)

Align the pin with the pin hole
Figure 11.18 Attached the Cell Panel (11)
(10) Connect the (OUT) tubing (1) and the (IN) tubing (9) compression screws to the
unions (16) and tighten them, while making sure check that the guide pin (14) enters
the pin hole (15) in the cell panel (11), then lightly tighten the cell panel retaining
screw (12) (See Figure 11.18).
78
(11) Connect the restrictor to the OUT side of the cell, set the pump to apply a pressure of
1.0 MPa using methanol, and then let fluid flow for 10 minutes. If a leak occurs
between a gasket (5) and the cell (6), further tighten the cell retaining screw (2) using
a wrench. Use gauze or laboratory tissues, to wipe away any methanol that has
leaked.
(12) Attach the cell mask (8) in the correct position using the two cell mask retaining
screws (13). (See Figure 11.19).
Correct Incorrect Incorrect
Figure 11.19 Positioning the Cell Mask (8)
(13) Attach the assembled flow cell to the FL-560. After checking that the pin on
the monochromator side is aligned with the pin hole of the cell panel (11), tighten the
lock screw.. Finally, tighten the cell panel retaining screw (12) (See Figure 11.20).
Tighten these screws first
Tighten this screw last
Figure 11.20 Attaching the Cell Panel (11)
79
11.2.6 Flow cell types
The FL-560 cell is coated with aluminum in order to use excitation and emission
light more efficiently. Scratches and peeling (up to approximately 0.5 x 1 mm) of the
aluminum coating will not cause a significant reduction in sensitivity. Cells that display
slight peeling of the aluminum coating may be used for some types of analysis (in which
detection sensitivity is not particularly high).
The aluminum coating on the cell will peel if the surface is rubbed by sharp objects, such
as tweezers. In addition, the aluminum coating may peel if the cell is soaked for long
periods in strong acids, strong salts, or organic solvents, or if solvent leaks onto the cell
for an extended period.
Table 11.1 Flow Cell Service Parts
Name Product code Part number Minimum

purchase
Cell replacement set
(Al coating) C458 6715-H311B 1 set
Note: The cell replacement set cons ists of one quartz cell and two gaskets.
11.3 What to do after leaks have occurred

11.3.1 Measures for flow cell
Causes and methods for dealing with leaks of the flow cell are described in Table 11.2.
Refer to Section 11.2 to disassemble, wash, and reassemble the flow cell.
When leakage is severe and salt has precipitated on the cell body or other parts, soak the
entire flow cell in warm water (approximately 40°C) before disassembly. Remove the salt
from the parts around the cell using a brush with soft bristles and then rinse the parts with
water. If any disassembled part has salt precipitate, use a brush to remove the salt and
then rinse the part with water.
80
Table 11.2 Causes and Methods for Dealing with Cell Leaks
Cause What to Do
Broken cell Replace the cell
Gasket deterioration Replace the
gasket
Clogging of the (OUT) tubing (1), (OUT) cell Remove the clog
holder (4), (IN) tubing (9), (IN) cell holder (7), or
unions (16)
Improper assembly Reassemble the
For example: cell
Parts have been assembled in incorrect
order
Improper fastening of cell retaining screw
Gauze or other material is adhering to a
gasket
11.3.2 Measures for monochromator
Remove the flow cell from the FL-560 and wipe up any solvent that has leaked in
the monochromator. When an organic solvent has leaked, wipe the solvent up using
gauze or laboratory tissue and let sit to dry. Buffer or salt solvents should be wiped up
using gauze or laboratory tissue that has been moistened with water.
Remove the cover on the bottom of the instrument, remove the receiving plate mounting
screws, then remove the receiving plate. Wipe up any solvent or salt on the receiving plate
or in the leak sensor section using a piece of gauze or laboratory tissue that has been
moistened with water (See Figure 11.21 and 11.22).
Install the flow cell and turn on power. If the TROUBLE LEAK IN CELL warning does not
appear on the display, the instrument may be used (See Note below).
When the warning appears even after properly wiping and drying, contact your local
KONIK distributor.
Note: K O NIK recommends that after carrying out the above procedures, the power be
turned off, the flow cell removed, and the instrument be left to dry for
approximately 24 hours.
81
Loosen these screws to
remove the cover.
Figure 11.21 Instrument Bottom
Receiving plate mounting screws Receiving plate
PCB
Since the "LEAK IN CELL" warning will occur again

This is the sensor if solvent or salt remain in this area,
be sure to clean this area sufficiently.
Removed receiving plate
Lead wire
Figure 11.22 Bottom View of the Instrument Inside and the Receiving Plate
82
11.4 Checking and inputting instrument constants
11.4.1 Situations requiring that the instrument constants be checked
The instrument constants must be checked in the following situations:
(1) The wavelength may have shifted.

For example, when peak height is very different compared to when previously used.
(2) Even though the wavelength has not shifted, the peak height is very different
compared to previous analyses under the same conditions.
(3) The wavelength calibration value has disappeared.

For example, when BACK UP ERROR occurs due to battery failure.
11.4.2 Location of Instrument Constants Label
The constants are written on a label inside the instrument (See Figure 11.23).
(1) Turn the power off and unplug the power cable.
(2) Remove the screws on the upper surface of the instrument case, and loosen the two
screws on the rear panel. Lift up the rear portion of the case and move it toward the
rear of the instrument to remove it (See Figure 11.24).
(3) Copy down the values on the label on the left side of the instrument.
(4) Attach the case and tighten the screws on the top and back of the instrument (See
Figure 11.24).
83
Instrument constants label
The values are recorded

on this label.
Note: S ince it is not necessary to input the AD J. of the H.T., this column should be blank.
Figure 11.23 Location of the Instrument Constant Settings Label
Remove this screw
The case can be removed after

the rear portion can be lifted.
Loosen these screws
Figure 11.24 Opening the Case
84
11.4.3 Input Method for Instrument Constants
Turn the power on and follow the procedure in Figure 11.25 to input the values recorded in
Section 11.4.2.
The settings listed in Table 11.3 are used in the following example. The user should input
the settings that correspond to the particular instrument being used.
Table 11.3 Example Instrument Constants
Instrument constant Settings

EX WL CONSTANTS 0 nm: 23, 546 nm: 558
EM WL CONSTANTS 0 nm: 19, 546 nm: 578
HT VOLTAGE CONST 1000: 654, (X100, X10: abbr. expl.) X1: 245.6
SLIT POSITION 18 nm: 15, (40 nm: 1750) 10 nm: 3450
NORM 0.0013 STD
300 500 1000 16

Monitor screen
[MONIT] [SHIFT] [MONIT]
RECALIB. PROGRAM
YES=ENT, NO=MONIT
Initial screen of calibration program
[MONIT] [EDIT/ENTER]
SELECT NO.
0:SET CONSTANTS
Input menu screen for constants
Figure 11.25.1 Method for Inputting Instrument Constants
85
Darkened areas designate flashing values
SELECT NO.
0:SET CONSTANTS
[EDIT/ENTER] EX wavelength constant input
EX WL CONSTANTS EX WL CONSTANTS
0= 20, 546=557 [EDIT/ENTER] 0= 20, 546=557

[2][3][EDIT/ENTER]
0= 23, 546=558 [5][5][8][EDIT/ENTER] 0= 23, 546=557
[ ] [ ]
EM WL CONSTANTS Input the EM wavelength constant in the same way as

the EX wavelength constant
0= 19, 546=578
[ ] [ ] H.T.constant input
HT VOLTAGE CONST HT VOLTAGE CONST

X1000=700 [EDIT/ENTER] X1000=700
HT VOLTAGE CONST [6][5][4][EDIT/ENTER]
X1000=654 Input X100, X10, and X1 in the same way
[ ] [ ]
Slit position constant input
SLIT POSITIONS [EDIT/ENTER] SLIT POSITIONS
18 nm= 20 18 nm= 20
SLIT POSITIONS [1][5][EDIT/ENTER]
18 nm= 15
Input 40nm and 10nm in the same way
Confirm that all input values Note: Use the up and down keys to scroll the screen
are correct and check the values.
[MONIT]
SET CONSTANTS When the [MONIT] key is pressed recalibration will be performed
automatically using each of the input values.
NOW SETTING" This message will be displayed for up to one minute while calibration
is being executed.
When recalibration is complete the input will appear.
Figure 11.25.2 Method for Inputting Instrument Constants
86
11.5 Cleaning the air filter
Two types of air filter are used with the FL-560. When an air filter is clogged, the
lamp cooling efficiency is reduced, contributing to faster lamp deterioration. Depending on
the environment, the air filters should be checked for dust and other materials
approximately once every month. When dust or other materials are clogging a filter, use
the procedures below to clean or replace the filter.
CAUTION:(1) If the power is turned on while an air filter is removed, dust will be
blown from the cooling fan into the lamp house.
(2) If an air filter that has been washed in water is attached without
sufficient drying and the power is then turned on, water will be
transferred to the lamp and will contribute to deterioration. Attach air
filters that have been washed in water only after they have been
allowed to dry sufficiently.
11.5.1 Cleaning the left side-panel air filter
(1) Turn the power off and remove the air filter (screen filter) (See Figure 11.26).
Driver
The air filter can be removed

by removing these four screws
Figure 11.26 Removal/Installation of the Side-panel Air Filter
(2) Using a brush, remove any dust that has collected on the filter. If this is difficult, wash
the filter in water.
(3) Re-attach the filter to the instrument after allowing the filter to dry sufficiently.
87
11.5.2 Cleaning the rear-panel air filter
(1) Turn the power off and remove the air filter (See Figure 11.27).
(2) Using a brush, remove any dust that has collected on the filter. If this is difficult, wash
the filter in water.
(3) Re-attach the filter to the instrument after allowing the filter to dry sufficiently.
Disassemble can be performed by pulling on the black frame (retainer).
Air filter (media)

Black mesh
Retainer
Guard
Figure 11.27 Removal/Installation of the Rear-panel Air Filter
88
11.6 Power Fuse Replacement
W AR NING: Make sure that the power cable is unplugged from the power input terminal
when replacing power fuses.
W AR NING: To prevent fire hazards or other possible accidents, use only replacement
fuses of the appropriate current values.
Table 11.4 Fuse capacity
Specified current Power source voltage

values
5.0 A 100 - 115 V
2.5 A 220 - 240 V
(1) To remove the fuse holder, use a flat-head screwdriver to simultaneously press down
and turn the holder in the counterclockwise direction (See Figure 11.28).
(2) Replace the fuse, and then replace the fuse holder by using the screwdriver to
simultaneously press down and turn the holder in the clockwise direction.
Note: W hen one fuse blows, replace both fuses.
An old fuse happens to blow accidentally, while an instrument operates properly. If the
new fuse blows immediately, there may be an instrument problem. Contact your local
KONIK distributor.
Figure 11.28 Replacing Power Fuses
89
12. Replacing Consumable Parts
The primary consumable parts of the FL-560 are listed in Table 12.1.
Table 12.1 Consumable Parts
Part Name Average Lifetime

Xenon lamp 1000 ~ 1300 hours
Light source condenser mirror 3000 ~ 5000 hours
Flow cell Dependent upon such as
used solvents, flow rate
or required sensitivity.
Note: The lifetime of the xenon lamp is defined as the period until the light intensity at
400 nm has decreased to 50% or less of the initial intensity, or until the lamp will
no longer light.
The average lifetime of a xenon lamp is approximately 1000 to 1300 hours. However,
since deterioration of the quartz portion is advanced in lamps that have been used for
more than 1000 hours, replacement should be performed soon after 1000 hours of use.
When performing analysis that requires high sensitivity (S/N), KONIK recommends that
the light source condenser mirror be replaced after 2000 to 3000 hours. However, if
sufficient analysis results are being obtained, the light source condenser mirror need not
be replaced even if the average lifetime has been exceeded.
When the light source condenser mirror must be replaced, contact your local KONIK
distributor.
When the instrument has been left unused for an extended period of time, the internal
optical elements may have deteriorated regardless of the total usage time. Use the
instrument carefully, and if problems occur, the instrument may require restoration
maintenance. Contact you local KONIK distributor.
KONIK recommends that every 1000 hours the whole lamp house unit should be replaced
to maintain the extra high sensitivity at replacement of light source for FP-2025. The whole
lamp house unit consists of the xenon lamp, the light source mirror, the glass plate and
the lamp hosing.
Optical alignment has already been done in the KONIK factory. In order to maintain the
light source energy it is necessary to replace not only the xenon lamp but also the light
source mirror. When replacing the whole lamp house unit, contact your local KONIK
distributor.
Note: W hen not using for extra high sensitivity analysis, sufficient performance of
sensitivity is obtained also by exchanging only a X enon lamp. However, if do not
changing the whole lamp hous e unit, specification of R aman scattering sensitivity
may not be obtained.
12.1 Flow cell part replacement

See Section 11.2.
90
12.2 Xenon lamp replacement
The life time of Xenon lamp is thousand hours. Over thousand hours operation causes not
only a reduction of energy and a increasing of fluctuation but also the quartz glass fatigue
is accumulated and the lamp bulb may be destroyed. If the lamp operation time exceeds
thousand hours, the used lamp must be replaced to new one with referring to below.
W AR NING: The lamp is made of quartz glass (hereafter, simply called glass), and
contains gas under high pressure (5-10 ⋅ atmospheric pressure, increasing
to 20-40 ⋅ atmospheric pressure when lit). Twisting, bending or other types of
stress may cause the lamp to explode, causing injury due to flying glass. In
particular, under no circumstances should you hold and twist both ends of the
lamp in opposite directions.
W AR NING: For safety reasons, when handling the lamp, use a protective mask, a thick,
long sleeve shirt, gloves, and other protective clothing.
W AR NING: When replacing the lamp, turn the power off, unplug the power cable from the
power socket, and wait approximately 15 minutes for the lamp to cool
sufficiently.
CAUTION:(1) Replace lamps that have been used for 1000 hours or more. Even
though the lighting may appear to be good, the glass portions of a
lamp that have been used for an extended period of time become
fatigued and are mechanically weak. Replacement should be
th
performed as soon after the 1000 hour as possible.
(2) Do not look directly at the light from the lamp without wearing
protective eyewear. Even scattered light contains eye-damaging
ultraviolet rays.
(3) Do not touch the glass of the lamp with bare hands. When the lamp
has been touched with bare hands or becomes dirty, clean the lamp
with alcohol-moistened gauze. If the lamp is lit while dust or
fingerprints are present on the quartz glass surface, the dust may be
burned and the transmittance of the glass may decrease, i.e., the light
intensity will drop. Moreover, the mechanical strength of the glass will
be decreased remarkably.
(4) Pay close attention to the polarity of the lamp. If the lamp is used after
being installed in the wrong direction, the electrodes will be
destroyed and the lamp will be useless.
(5) Be sure the cover of the lamp housing and instrument case are
properly attached before lighting the lamp (except during lamp
position adjustment).When the lamp housing or instrument case is
not properly attached, the lamp temperature may become abnormally
91
high and there is dauger of electrical shock from accidental contact
with the high voltage power source.
(6) Place the old lamp in the case in which the new lamp was packaged
and store the case in a safe location. If a case is not available, wrap
the old lamp securely in shock absorbing materials and store in a
safe place.
(7) To process used lamps, wrap the lamp securely in thick cloth or
similar material, strike the bundle with a hammer or other object of
similar mass, and process as hazardous material. If performing this
procedure is impossible, carefully package the lamp and contact your
local KONIK distributor.
12.2.2 Lamp replacement
W AR NING: When replacing the lamp, use only lamps with the UXL - 159H or 5330-0086
designation. Lamp explosion or lighting abnormalities may occur if other
lamps are used. When replacing the lamp, confirm that UXL-159H or 5330-
0086 is imprinted on the electrode portion of the lamp (See Figure 12.1).
CAUTION: Although the UXL-159 lamp has conventionally been used with the
KONIK 820-FP and 821-FP fluorescence detectors, this lamp cannot be
used with the FL-560. The UXL-159 lamp may explode or cause
lighting abnormalities if used in the FL-560.
Take special care when replacing the lamp because the UXL-159 and
the UXL-159H (5330-0086) lamps are the same shape.
Note: The UX L-159H (5330-0086) may be used in the 820-FP and 821-FP fluorescence
detectors.
Make sure that UXL-159H or 5330-0086 is written here.
Figure 12.1 Location of Lamp Model Number
To replace the lamp, follow the procedure described below:
(1) Turn the power off and unplug the power cable.
(2) Allow the instrument parts to cool.
W AR NING: The lamp temperature is several hundred degrees Celsius immediately after
being turned off. Be sure to allow the instrument sit for 15 minutes or more
before performing these procedures.
92
(3) Put on a heavy long-sleeved shirt, protective gloves, and a protective mask.
(4) Open the fan door (See Figure 12.2).
Figure 12.2 Opening/Closing the Fan Door
(5) Insert the protective plate (See Figure 12.3).
Note: The protective plate provides protection against lamp explosion and is a standard
access ory of the FL-560.
Protective plate
Figure 12.3 Attachment/Removal of the Protective Plate
93
(6) Remove the screws securing the electrode holder, grasp the electrode holder, and
gently remove it while the lamp is still attached (See Figure 12.4).
Electrode holder (1)
Remove the screws securing the electrode holder and

remove the holder and lamp together
Top view
Approx. 10
Approx. 5
Side view
Figure 12.4 Lamp Removal/Installation
(7) Refer to Fig 12.5, remove the electrode holders from the old lamp and attach them to
the new lamp.
W AR NING: When removing and attaching electrode holders, do not hold both ends of the
lamp and twist, as this may destroy the lamp.
94
Note: D o not turn the screw in the middle of electrode holder (1). This screw is for lamp
pos ition adjustment.
This is the lamp position

adjustment screw.
Do not turn this screw at this
Electrode time.
holder (1)
Electrode To attach or remove the lamp

holder (2) from electrode holder (2), hold
the lamp with a gloved hand and
Anode side turn the electrode holder(2)
Counter clockwise (clockwise)
when remouval (attachment)
To attach or remove the lamp

from electrode holder (3), hold
the lamp with a gloved hand and
Electrode turn the electrode holder(3)
holder(3)
Cathode side
Removal Attachment
Figure 12.5 Lamp Electrode Attachment/Removal
(8) Adjust the lamp direction (See Figure 12.6). When the two holes for the screws that
secure the electrode holder (1) are aligned vertically, hold electrode holder (1) and
turn electrode holder (2) so that the protrusion is horizontal. Then turn the trigger wire
so as to be positioned just above the protrusion.
95
Trigger wire
Protrusion
Screw holes Electrode holder (1) Electrode holder (2)

Up Trigger wire
Horizontal line
Protrusion
Down Turn these sections to achieve the arrangement
shown at left.
View from arrow position
Figure 12.6 Positional Relationship between the Lamp Protrusion and the Trigger Wire
W AR NING: When inserting the lamp housing, be careful not to hit the lamp on the lamp
housing as doing so may destroy the lamp.
Note: W hen attaching the lamp (along with the electrode holders) to the lamp housing,
secure the lamp such that the trigger wire is in position above the lamp protrusion.
(9) Grasp electrode holder (1) and insert the lamp (along with the electrode holders) into
the lamp housing. If the lamp does not slide in smoothly, do not force it. Instead, find
the correct insertion direction by slowly moving the insertion end (electrode holder (3))
up, down, left, and right. Refer to Figure 12.4 for the correct insertion angles.
Electrode holder (3) is made to slide smoothly into the lamp housing hole (which is
difficult to see from outside the instrument, Refer to Figure 12.7).
96
Cathode terminal (Gold-colored)
Internal view of the lamp housing

from the rear of the FP-1520
when the lamp is removed.
Electrode holder (3) is inserted into this hole.

Sleeve (Teflon)
Figure 12.7 Electrode Holder (3) Insertion Hole
(10) Tighten the securing screws (See Figure 12.4).
12.2.3 Lamp position adjustment
Lamp position adjustment is required after lamp replacement. The adjustment procedure
is described below.
W AR NING: When the fan door is open, do not turn the power on without protective
plate. Injury may result if the lamp is destroyed without protective plate.
C aution (1): R emove the protective plate only after completing lamp position adjustment
and turning the FL-560 power off.
C aution (2): P erform lamp pos ition adjustment quickly after lighting the lamp (turning the
power on). Lamp deterioration will be accelerated if the lamp is lit for 20
minutes or more while the fan door is open.
(1) Confirm that the protective plate is inserted into the lamp housing.
(2) Plug the power cable into the FL-560 and turn the power on.
97
(3) When the monitor screen appears, set the excitation wavelength to 400 nm and the
fluorescence wavelength to 500 nm, and then press [SHIFT][1]. The screen in Figure
12.8 will be displayed.
EX : X.XXX V
EM: Y.YYY V
Figure 12.8 Displaying the Preamplified Output
(4) Turn the position adjustment screw until the output signal for excitation x.xxx is
maximized (See Figure 12.9).
Lamp position adjustment screw
Do not turn these screws.
Figure 12.9 Adjusting Lamp Position
(5) Turn the power off.
(6) Remove the protective plate.
(7) Close the fan door and secure with the holding screws (See Figure 12.2).
(8) Turn the power on and check that the fan is turning.
(9) When the monitor screen appears, set the lamp-use time to 0 by pressing [SHIFT][7].
(Refer to Section 5.8).
98
12.2.4 After replacing the lamp
Two types of air filters are used in the FL-560. When an air filter is clogged, the
lamp cooling efficiency is reduced, contributing to faster deterioration. Depending on the
operating environment, the air filters should be checked for dust and other materials
approximately once every month. When dust or other materials are blocking a filter, refer
to Section 11.5 and clean or replace the filter.
12.3 List of consumable parts

Table 12.2 List of Consumable Parts
Item Name Part number Minimum Description of

purchase Contents
Gasket 6715-H301B 1 set IN and OUT (1 each)
Cell replacement set 6715-H311B 1 set Cell and gasket
(Al coating)
Xenon lamp 6715-H571A 1 set Lamp and air filter
replacement set
99
13. Performance Test
13.1 Checking wavelength accuracy and wavelength calibration

13.1.1 Definition of wavelength accuracy
Although the primary function of the FL-560 is not the measurement of spectra or
the precise evaluation of wavelength and fluorescent intensity, a certain degree of
wavelength accuracy is required in order to compare data between systems. Moreover,
large deviations in wavelength may result in reduced reliability for analysis results.
Periodic confirmation of wavelength accuracy is therefore recommended.
When confirming wavelength accuracy, use a recorder that has been calibrated in
accordance with public standards.
13.1.1.1 Wavelength accuracy of the fluorescence (EM) monochromator:
Wavelength accuracy is expressed as a function of the error between the measured

wavelength and the true wavelength. Upon measurement using of the fluorescence
monochromator, a light source is placed at the flow cell section. After spectrum
measurement via scanning monochromator, compare the true (known) wavelength of the
light source to the measured wavelength.
There are two methods for confirming the wavelength accuracy of the EM monochromator.
One method involves attaching a jig cell, containing a low-pressure mercury lamp, to the
flow cell and measuring the line spectrum of the mercury lamp. The other method involves
inserting holmium glass, built into the fluorescence monochromator of the F-560,
into the fluorescent light path and then measuring the absorption spectrum.
R eference values of wavelength accuracy for determining the acceptance criteria for the
fluorescence monochromator.
Instrument specification: within ±2 nm

R ecommended operating standard: within ±4 nm
Note: W hen using holmium glass for wavelength calibration, the absorption wavelength
variation of the glass itself, approximately ±1 nm, must also be considered (due to
the certified absorption wavelength variation and imperfect flatness of the base
level near the holmium absorption peak). W hen using holmium glass for
wavelength calibration, this variation must be taken into account for determining
the acceptance criteria.
13.1.1.2 Wavelength accuracy of the excitation (EX) monochromator:
Wavelength accuracy is expressed as a function of the error between the measured

wavelength and the true wavelength. Upon measurement using the fluorescence
monochromator, sample having a known emission spectrum is placed in the flow cell.
100
After spectrum measurement via scanning monochromator, compare the true (known)
wavelength of the sample to the measured wavelength.
Confirmation of the excitation wavelength accuracy is performed using the Raman
scattering of water. When excited by light having a wavelength of 350 nm, the Raman
spectrum peak of water is observed at 397 nm.
The wavelength of the EX monochromator is therefore set to 350 nm and the Raman
spectrum of water is measured. The peak wavelength from the spectrum is determined
and the deviation from 397 nm is calculated.
Note: The wavelength accuracy of the E X monochromator must be confirmed after

confirming the wavelength accuracy of the fluorescence monochromator.
R eference values for determining the acceptance criteria for the excitation monochromator
Instrument specification: within ±2 nm

R ecommended operating standard: within ±4 nm
13.1.2 Wavelength accuracy confirmation and calibration for EM monochromator
13.1.2.1 Confirmation of fluorescence monochromator wavelength accuracy using a

low-pressure mercury lamp
<Test method>
Attach a jig cell containing a low-pressure mercury lamp in the flow cell section and scan
the EM monochromator. Measure and record the intensity spectrum of the mercury lamp.
Read the 254 and 546 nm peak wavelengths from the recorded intensity spectrum.
Repeat the above operations three times and calculate the average value for each
wavelength. Confirm that the difference between the averages and the true values is
within the limits of the acceptance criteria.
<Required equipment>
Jig cell
Recorder
<Test preparations>
(1) Remove the flow cell and attach the jig cell to the flow cell section.
(2) Connect a recorder to the REC terminal on the back of the FL-560.
Note: W avelength marker signals will be output from the R E C terminal at every 100 nm.
R ead the wavelengths based on the marker signals. D o not mistakenly connect
the recorder to the INT terminal, as no wavelength marker signal is output from
this terminal.
(3) Turn on the FP-2020/2025 power and light the low-pressure mercury lamp.
Note: W ait at least 30 minutes after the power has been turned on and 5 minutes after
the mercury lamp has been lit before starting the test.
101
<Test procedure>
Note: R efer to the operations manual for a more detailed description of operations. The
following is only a brief outline of the detailed explanation provided in the
operations manual.
(1) Set the fluorescent bandwidth to 18 nm.

(2) Follow the procedure described in Figure 13.1 to measure the intensity spectrum of
the low-pressure mercury lamp. Change the memory number and repeat the
measurement three times.
300 500 1000 16

RECALIB. PROGRAM Initial screen of
calibration program
YES=ENT, NO=MONIT
Input menu
SELECT NO. screen for
constants Fluorescence spectrum
0:SET CONSTANTS Change the memory number and
repeat the measurement three times.
[ ] [ ]
[MONIT]
SELECT NO. EM SPECTRUM MEASURE Memory number
selection screen
1:EM WL. CHECK [EDIT/ENTER] SET MEMORT NO. X
[ ] [ ] EM wavelength
confirmation [X][EDIT/ENTER]
menu screen Spectrum
EM SPECT. MEASURE measurement
Menus 2 through 14 will appear start screen
Do not open these menus PUSH PRGM RUN
(see Note). [PRGM RUN]
EM SPECT. MEASURE Screen during

spectrum
RUNNING measurement
Figure 13.1 Fluorescence Spectrum Measurement
Note: Menus 2 through 14 of the calibration program are intended for use strictly by
service engineers. D o not open these menus . The ins trument will not operate
properly if these menus are opened and the settings are changed.
(3) Output the three measured spectrums to the recorder.
102
(4) Read the wavelengths for the 254 and 546 nm peaks from each spectrum. When
these peaks are broad, draw lines as described in Figure 13.2, find the intersection
point, and use this point as the peak wavelength.
Intensity Peak wavelength
Wavelength
Figure 13.2 Reading Peak Wavelength for Mercury Lamp
(5) Calculate the average values for peaks at 254 and 546 nm.
(6) Calculate the difference between the average values and the true values.
Average - 254 = d1
Average - 546 = d2
Note: The values for d1 and d2 are required for wavelength calibration.
(7) Operation is normal if the values for d 1 and d2 fall within the limits of the acceptance
criteria. When the acceptance criteria are exceeded, refer to Section 13.1.2.3 and re-
calibrate the wavelength.
103
13.1.2.2 Confirmation of fluorescence monochromator wavelength accuracy using
holmium glass
<Test method>
Insert holmium glass into the light path of the EM monochromator. Measure the EM
spectrum and output the results to a recorder. Read the absorption peaks for holmium
glass from the EM spectrum and calculate the difference between the measured
wavelengths and the true wavelengths. Repeat the procedure described above three
times and calculate the average value for each wavelength. Confirm that the differences
between the averages and the true values are within the limits of the acceptance criteria.
Recorder
Note: Holmium glass is included as standard equipment in the FL-560.
<Test preparations>
(1) Attach a standard flow cell (16 µL capacity) to the FL-560, and set a flow cell
containing high-purity water.
R ead the wavelengths based on the marker signals. D o not mistakenly connected
this terminal.
(3) Turn on the FL-560 power.
Note: W ait at least 30 minutes after the power has been turned on before starting the
test.
<Test procedure>
Refer to the operation manual for a more detailed description of operations.
(1) Wavelength range setting

Press [SHIFT][CLEAR] to display the wavelength range screen, and then select NOT
RESTRICTED (0 ~ 900 nm).
(2) Place the holmium glass in the light path

Press [SHIFT][9] to display the spectrum bandwidth screen then select HOLMIUM.
(3) Set the measurement parameters as follows:

Ex wavelength: 0nm
GAIN: ⋅ 10 or ⋅ 100
ATTEN: ⋅ 64
104
(4) Spectrum measurement parameter settings
Press [SCAN][0] to display the scan parameter settings screen, then select RNG:
STD and SPD: 2.
(5) Measure the EM spectrum (holmium absorption spectrum).
(6) Output the measured spectrum to the recorder.
(7) Read the wavelength for the valley at 361 nm (See Figure 13.3). When the valley
cannot be determined, change the GAIN and ATTEN settings and measure the EM
spectrum again.
Holmium absorption wavelength
Fluorescent
intensity
Wavelength
Figure 13.3 Reading Absorption Wavelength for Holmium Glass
(8) Repeat the measurement and recording procedures three times.
(9) Calculate the average wavelength for the valley.
(10) Calculate the difference between the average wavelength and the true value. When
the acceptance criteria are exceeded, refer to Section 13.1.2.3 and re-calibrate the
wavelength.
Average value - 361 = d3
Note: The value d3 is required for wavelength calibration.
(11) Operation is normal if the value for d3 falls within the limits of the acceptance criteria.
(12) Be sure to return all instrument settings to the settings that were in use before
checking the wavelength. Repeat the operations described in Steps (1) and (2) to
return the FL-560 to normal operations.
C AUTIO N: The instrument will not operate properly if the wavelength range is not
returned to the previous settings and the holmium glass is not removed from
the light path.
105
13.1.2.3 Wavelength calibration for the EM monochromator
(1) Press [SHIFT][MONIT] to display the calibration program screen.
(2) Use the procedure described in Figure 13.4 to display the input screen for EM
wavelength constants.

300 500 1000 16

calibration program
YES=ENT, NO=MONIT
SELECT NO. Input menu screen
for constants
0:SET CONSTANTS
[EDIT/ENTER]
EX WL CONSTANTS
0= 20, 546=557
[ ] [ ] Shaded areas designate flashing values.
EM WL CONSTANTS EM WL CONSTANTS
0= 19, 546=578 [EDIT/ENTER] 0= 19, 546=578

[2][3][EDIT/ENTER]
EM WL CONSTANTS EM WL CONSTANTS
[5][5][8][EDIT/ENTER]
0= 23, 546=558 0= 23, 546=578
[MONIT]
SET CONSTANTS When the [MONIT] key is pressed, recalibration will be

performed automatically using each of the input values.
NOW SETTING This message will be displayed for up to one minute while
calibration is being performed. When recalibration is complete,
the Input menu screen will appear.
Figure 13.4 Method for Inputting EM wavelength constant
(3) Input the EM wavelength constant found by performing the following calculation:
When the wavelength accuracy was checked using the intensity spectrum of the
mercury lamp, described in Section 13.1.2.1, input new values for 0 and 546.
New 0 value setting: Value Setting before recalibration + (2 ⋅ d 1 - d2)
New 546 value setting: Value Setting before recalibration + 2 ⋅ (d2 - d1)
When the wavelength accuracy was checked using the absorption spectrum of
holmium glass, described in Section 13.1.2.2, change the setting for 0 only (leave the
546 setting as it is).
New 0 setting: Value Setting during spectrum measurement + d3
106
(4) Press the [MONIT] key to automatically calibrate the wavelength and return to the
constants input menu screen.
(5) Press the [MONIT] key again to return to the monitor screen.
(6) Follow the procedure described in Section 13.1.2 to check the wavelength. If the error
exceeds the limits of the acceptance criteria, recalibrate the wavelength.
13.1.3 Wavelength accuracy confirmation and calibration for the EX

monochromator
13.1.3.1 Confirmation of EX monochromator wavelength accuracy
<Test method>
After confirming the wavelength accuracy of the EM monochromator, use the 397 nm
peak on the Raman spectrum for water, excited at 350 nm, to confirm the wavelength
accuracy of the EX monochromator.
Measure the Raman spectrum for water and output the results to a recorder. Read the
peak at 397 nm from the recorded spectrum and calculate the difference between the
measured peak and the true peak. Repeat the above-described procedures three times
and calculate the average peak value. Confirm that the difference between the average
value and the true value is within the limits of the acceptance criteria.
Recorder
<Test preparations>
this terminal.
Note: W ait at least 30 minutes after the power has been turned on before starting the
test.
(3) Connect the HPLC pump to the flow cell and flow high-purity water through the cell at
1 mL/min.
<Test procedure>
Ex wavelength: 350 nm
GAIN: ⋅ 1000
ATTEN: ⋅ 128
Fluorescent bandwidth: 18 nm
107
(2) Spectrum measurement parameter settings.
Press [SCAN][0] to display the scan parameter screen then select RNG: STD and
SPD: 2.
(3) Measure the EM spectrum (Raman spectrum for water).
(4) Output the measured Raman spectrum to the recorder.
(5) Read the wavelength for the peak at 397 nm (See Figure 13.5).
Peak wavelength of Raman scattering
Fluorescent
Intensity
400nm marker
Wavelength
Figure 13.5 Reading the Peak Top of Raman Scattering
(6) Repeat the measurement and recording three times.
(7) Calculate the average wavelength for the peaks.
(8) Calculate the difference between the average wavelength and the true wavelength of
the peak at 397 nm.
Average - 397 = d4
Note: The value d4 is required for wavelength calibration.
(9) Operation is normal if the value for d4 falls within the limits of the acceptance criteria.
13.1.3.2 Wavelength calibration for the EX monochromator
(1) Press [SHIFT][MONIT] to display the calibration program screen.
108
(2) Use the procedure described in Figure 13.6 to display the input screen for EX
wavelength constants.
300 500 1000 16

calibration program
YES=ENT, NO=MONIT
SELECT NO. Input menu screen
for constants
0:SET CONSTANTS
[EDIT/ENTER] Shaded areas designate flashing values.
0= 20, 546=557 [EDIT/ENTER] 0= 20, 546=557
[2][3][EDIT/ENTER]
[EDIT/ENTER]
0= 23, 546=557 0= 23, 546=557
[MONIT]
SET CONSTANTS When the [MONIT] key is pressed a recalibration will

automatically be executed using each of the input values.
NOW SETTING" This message will be displayed for up to 1 minute while
calibration is being executed. When recalibration is complete
the input menu screen will appear.
Figure 13.6 Method for Inputting EX wavelength constant
(3) Input the 0 constant for the EX wavelength obtained using the following calculation
(do not change the 546 setting):
New 0 value: Value before recalibration - d4
(4) Press the [MONIT] key to automatically calibrate the wavelength and return to the
constants input menu screen.
(5) Press the [MONIT] key again to return to the monitor screen.
(6) Follow the procedure described in Section 13.1.3.1 to check the wavelength. If the
error exceeds the limits of the acceptance criteria, recalibrate the wavelength.
13.2 Confirmation of minimum detectable amount

13.2.1 Meaning of minimum detectable amount
There are several definitions for minimum detectable amount with respect to the HPLC
fluorescence detector. In order to allow good repeatability and ease of use, KONIK
109
recommends the definition according to the S/N ratio for the Raman light of water. The
S/N ratio is expressed as the ratio between the peak height for the Raman light of water
and the noise level at the peak top wavelength.
<Reference values for determining acceptance criteria for minimum detectable amount>
Instrument specification: 320 (FL-560)
Instrument specification: 450 (FP-2025)
Instrument specification: 150 (FL-560, 5 µL semi-microflow cell)
Recommended operating standard: 150 < (FL-560)
Recommended operating standard: 250 < (FP-2025)
Recommended operating standard: 80 < (FL-560, 5 µL semi-microflow cell)
13.2.2 Test method for minimum detectable amount
<Test method>
The ratio between the peak height for the Raman spectrum of water, excited at 350 nm,
and the noise level at the peak top wavelength is measured. Measurement is performed
using high-purity water flowing through the flow cell.
(1) The Raman spectrum is output to a recorder and the peak height for the Raman light
is then read.
(2) The wavelength of the EM monochromator is set to the peak top for Raman light and
the time variation is recorded. Recording is performed for 15 minutes. Public
standards.
Recorder that has been calibrated in accordance with public standards.
<Test method>
this terminal.
Note: wait at 1 east 30 minutes after the power has been turned on before starting the
test .
(3) Connect the HPLC pump to the flow cell and flow high-purity water through the cell at
1 mL/min.
<Test procedure>
EX wavelength: 350 nm
110
GAIN: ⋅ 1000
ATTEN: ⋅ 128
(2) Press [SCAN][0] to display the scan parameter screen then select RNG: STD and
SPD: 2.
(3) Measure the EM spectrum (Raman spectrum for water).
(4) Output the measured Raman spectrum to the recorder.
(5) Read the peak height for the Raman scattering of water using the neighboring line
method (See Figure 13.7). This height corresponds to S (signal).
When the Raman scattering peak is too large or too small with respect to the scale of
the recorder, adjust the ATTEN setting accordingly.
Peak wavelength of Raman scattering
Fluorescent
intensity
400nm marker
Raman scattering peak height
Wavelength
Figure 13.7 Reading Peak Height for Raman Scattering
(6) Confirm that the baseline has stabilized, set the EM wavelength to the peak
wavelength of the Raman spectrum, and record the time variation for 15 minutes
under the following conditions. When the noise is too large or too small with respect to
the scale of the recorder, adjust the ATTEN setting accordingly.
EX wavelength: 350 nm
EM wavelength: Wavelength at the peak top for Raman scattering
GAIN: ⋅ 1000
ATTEN: ⋅ 4
RESPONSE: STD
111
(7) Read the noise (peak-to-peak) from the baseline record.
Fluorescent
intensity
Noise
0 36 91215
Time (min)
Figure 13.8 Reading Noise Width of Raman Scattering
(8) Calculate the S/N value.

As an example, the calculation with S = 50 (ATTEN = 128) and N = 6 mm (ATTEN =
4) would be as follows:
S/N = (50 x 128)/(6 x 4) = 267
112

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MODEL FL 560

(1) Safety symbols

WARNING A WARNING indicates an potentially hazardous situation which, if not

Do not proceed beyond a WARNING or CAUTION notice until you

Note A Note provides additional information to aid the operator in obtaining

Pressurized, hazardous solvents are used in high-performance liquid chromatography.

Connecting the outlet of this detector to another detector or tubing with

(3) Warning Labels

(1) Lamp Replacement Warning Label (P/N: 0822-0078A)

MAY CAUSE BURSTING AND OTHER

Figure 1 Front View of the FL-560

(2) Danger Warning Label (P/N: 0822-0035A)

(4) Attention Warning Label (P/N: 0822-0036A)

(5) Fuse and Ground Warning Label (P/N: 0822-0163A)

The Xenon lamp is located below this label. Do not touch

(7) Lamp Replacement Warning Label (P/N: 0822-0079A)

Refer to the operations manual for details regarding lamp replacement.

(3) Xenon lamp warning label

(4) Attention warning label

(2) Danger warning label

(6) Hot surfaces warning label

(7) Lamp replacement warning label

(5) Fuse and Ground Warning Label

Figure 3 Figure of the side view

Figure 4 Grip Locations for Carrying the Instrument

EN55011 ---- ”Limits and Methods of Measurement of Radio Interference

EN50082-1 ---- Electromagnetic compatibility ---- Generic immunity standard Part 1:

IEC61000-4-2 ---- “Electromagnetic compatibility for industrial-process measurement

IEC61000-4-3 ---- “Electromagnetic compatibility for industrial-process measurement

IEC61000-4-4 ---- “Electromagnetic compatibility for industrial-process measurement

IEC61000-3-2: 1995 + Amd.14:2000 ---- “Electromagnetic compatibility: Limits for

To ensure safe operation, the following recommendations should be observed:

(3) Operate the instrument under a temperature range of 10 ∼ 30°C.

(7) Avoid vibrations caused by vacuum pumps, electric motors, processing

(2) Software prohibitions:

(4) Unauthorized copying of this manual is prohibited.

This warranty does not apply to defects as a result of the following:

(1) USE FOLLOWING IMPROPER OR INADEQUATE INSTALLATION.

(2) IMPROPER OPERATION.

(3) MOVEMENT, MODIFICATION, OR REPAIR BY PERSONS OTHER THAN

(5) INORDINATELY RAPID DETERIORATION DUE TO THE USE OF CORROSIVE

(6) NATURAL DISASTERS SUCH AS FIRES, WATER DAMAGE, OR

In addition, this warranty does not cover:

CONSUMABLE PARTS OR PARTS THAT HAVE A SEPARATE WARRANTY OR

KONIK Corporation, 2003

1.1 Product Description

1) Compact, high-efficiency monochromator that incorporates a holographic concave

Monochromator: Holographic concave diffraction grating

Spectrum bandwidth: Excitation side: 18 nm

Excitation side: Photodiode

Sensitivity: S/N for the Raman peak of water:

Measurement range: 10 steps in total: 1, 2, 4, 8, 16, 32, 64, 128, 256,

Gain: x1000, x100, x10, and x1

Response: Fast, standard, slow, and digital filter methods

Signal processing: Digital processing by A/D and D/A converters

Output: Recorder output: 10 mV/FS (polarity change is

Input: Marker input, autozero input and Program reset run

Program functions: Time programmability for EX wavelength, EM

Wavelength scan function: Excitation spectrum and emission spectrum

Diagnostic test function: Memory (ROM and RAM), DC power, EX energy

Lamp off timer: 99.9-hour maximum

Calibration function: EX and EM wavelength calibration

Rated power: AC100 240V±10V%, 50/60Hz, 425 VA