Metrohm 817 User Manual
817 Bioscan
Bioscan817
Bioscan
CH-9101 Herisau/Switzerland
E-Mail info@metrohm.com
Internet www.metrohm.com
Instructions for Use
8.817.1003
CH-9101 Herisau/Switzerland
E-Mail info@metrohm.com
Internet www.metrohm.com
817 Bioscan
Instructions for Use
8.817.1003 09.2001 / up
Contents
Contents
1 Introduction............................................................................................ 1
1.1 Instrument description............................................................................. 1
1.2 Parts and controls .................................................................................... 2
1.3 Information about the Instructions for Use .......................................... 8
1.3.1 Organization.................................................................................. 8
1.3.2 Notation and pictograms.............................................................. 9
1.4 Safety notes ............................................................................................. 10
1.4.1 Electrical safety........................................................................... 10
1.4.2 General precautionary rules ....................................................... 10
2 Installation........................................................................................... 11
2.1 Setting up the instruments.................................................................... 11
2.1.1 Packaging ................................................................................... 11
2.1.2 Check.......................................................................................... 11
2.1.3 Location ...................................................................................... 12
2.1.4 Arrangement of the instruments................................................. 12
2.2 Mains connection ................................................................................... 12
2.2.1 Setting the mains voltage ........................................................... 12
2.2.2 Fuses .......................................................................................... 13
2.2.3 Mains cable and mains connection ........................................... 14
2.2.4 On/Off switching ......................................................................... 14
2.3 Capillary connections ............................................................................ 15
2.3.1 Connection of the 6.5324.000 Bottle Rack (option)................... 15
2.3.2 Connection of the pulsation absorber........................................ 15
2.3.3 Connection of the injection valve ............................................... 16
2.3.4 Mounting the column.................................................................. 16
2.3.5 Connection of the flow cell ......................................................... 17
2.3.6 Connecting the waste tubing...................................................... 18
2.3.7 Rinsing the tubing....................................................................... 19
2.3.8 Rinsing the column ..................................................................... 19
2.4 System for sugar analysis – MIC 8 ...................................................... 20
2.4.1 Electrical connections................................................................. 20
2.4.2 Settings in «IC Net 2.1» ............................................................. 21
2.4.3 Connecting a sample changer ................................................... 22
2.5 System for the analysis of anions and sugars – MIC 9.................... 24
2.5.1 Electrical connections................................................................. 24
2.5.2 Capillary connections ................................................................. 25
2.5.3 Configuration in «IC Net 2.1» ..................................................... 27
2.6 Preparing the 817 Bioscan for the analysis ....................................... 32
2.6.1 System switch-on........................................................................ 32
2.6.2 Start pump .................................................................................. 32
2.6.3 Set operating mode.................................................................... 33
817 Bioscan
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Contents
3 Operation............................................................................................... 35
3.1 Handling the 817 Bioscan ..................................................................... 35
3.2 Operation using «IC Net 2.1» ................................................................ 36
3.2.1 817 Bioscan icon.........................................................................36
3.2.2 817 Bioscan window ...................................................................37
4 Basic principles............................................................................. 47
4.1 Introduction ............................................................................................. 47
4.2 Measuring conditions ............................................................................ 48
4.3 Pulsed amperometric detection........................................................... 49
4.3.1 Optimization of the PAD parameters ..........................................49
4.4 Optimization of the measuring potential ............................................ 51
5 Notes – Maintenance – Faults....................................... 55
5.1 Practical notes on ion chromatography ............................................. 55
5.1.1 Separating columns ....................................................................55
5.1.2 High-pressure pump ...................................................................56
5.1.3 Eluents .........................................................................................57
5.1.4 Connections ................................................................................57
5.2 Maintenance and servicing................................................................... 58
5.2.1 General information .....................................................................58
5.2.2 Shutdown.....................................................................................58
5.2.3 Cleaning the working electrode ..................................................59
5.2.4 Changing separating columns....................................................60
5.3 Faults and malfunctions ........................................................................ 61
5.3.1 Malfunctions and their rectification .............................................61
5.4 Instrument test with the dummy cell ................................................... 63
5.5 Validation / GLP ...................................................................................... 64
6 Appendix................................................................................................. 65
6.1 Technical data......................................................................................... 65
6.2 Standard equipment............................................................................... 69
6.3 Warranty and conformity....................................................................... 71
6.3.1 Warranty.......................................................................................71
6.3.2 EU declaration of conformity.......................................................72
6.3.3 Certificate of conformity and system validation ..........................73
6.4 Index ......................................................................................................... 75
II
817 Bioscan
Contents
List of illustrations
Fig. 1: Front of the 817 Bioscan................................................................................... 3
Fig. 2
: Rear of the 817 Bioscan.................................................................................... 4
Fig. 3
: Interior of the 817 Bioscan (with permanently attached accessories) .. 6
Fig. 4
: Setting the mains voltage ............................................................................... 13
Fig. 5
: Connections at the injection valve of the 2.812.0010 Valve Unit ......... 16
Fig. 6
: Connections at flowcell ............................................................................... 17
Fig. 7
: Connecting up a simple modular IC system
with the 817 Bioscan – MIC 8
Fig. 8
: Connecting up a simple MIC 8 system
including a sample changer
Fig. 9
: Connections with a 817 Bioscan as second detector
with a modular IC system – MIC 9
..................................................................... 20
....................................................................... 22
............................................................. 24
Fig. 10
: Connections of injection valves A and B in the IC Separation
Center (2.733.0120)
Fig. 11
: Potentials applied during pulsed amperometric detection (PAD). ...... 49
Fig. 12
: Example of a hydrodynamic voltammogram of a
..................................................................................... 26
substance (A) with additional presentation of the
measured values for the pure eluent (B)
Fig. 13
: Example of a scan of a substance (A) with additional
presentation of the scan of the pure eluent (B)
.................................................. 51
...................................... 52
817 Bioscan
III
Contents
IV
817 Bioscan
1.1 Instrument description
1 Introduction
1.1 Instrument description
The 2.817.0010 Bioscan is a PC-controlled measuring instrument for
the sensitive analysis of carbohydrates by ion chromatography using
pulsed amperometric detection. The compact housing of the 817 Bioscan contains several IC system components:
• Detector – flow-through cell with three-electrode arrangement for
amperometric detection in the Pulse, DC and Scan modes.
• Column compartment – the perfect insulation of the housing cre-
ates not only stable thermal conditions for the separating columns,
but also shields the system from electromagnetic interference; in
addition to the column it also contains the detection cell, pulsation
absorber and preheating capillary.
• Oven – amperometric determinations require extremely stable
thermal conditions. The built-in column compartment oven ensures
that all important components can be set exactly to a temperature
from 10°C above room temperature to 60 °C with a stability of
0.1°C.
• All components that come into contact with the eluent and the
sample, except the electrodes, are metal-free.
• Signal converter – the 817 Bioscan contains its own analog/ digi-
tal converter for the detector signal. This means that complete control of the instrument is possible from a PC via an RS232 interface.
Together with the 709 IC Pump and the 812 Valve Unit, the 817 Bio-
scan forms a complete IC system for carbohydrate analysis. Operation is via a PC connected to the RS232 interface and uses the in-
cluded «IC Net 2.1» control and evaluation program. This software
fulfills all the demands which are placed today on a modern integration
software: 1-point or multi-point calibration, internal or external standard,
selectable algorithms for non-linear calibration, numerous Integration
modes with selectable parameters and integration events, various peak
recognition methods, peak editor, free scaling, superimposition of several chromatograms, post-treatment of chromatograms, high-performance GLP-conform report generator with output interfaces for monitor,
printer and external databases.
817 Bioscan
1
1 Introduction
817 Bioscan
2
1.2 Parts and controls
1.2 Parts and controls
Bioscan817
Bioscan
1 2 3
: Front of the 817 Bioscan
Fig. 1
Door to interior 3 Feed through for tubing
1
e.g. connection of injection valve and preheating capillary or connection to waste
2 Feed through for tubing
e.g. connection of 709 IC Pump and
pulsation absorber
817 Bioscan
3
1 Introduction
4
4
4
4
5
6
4
4
7
8
18
17
9
16
4
10
11 14
12 13 15
Fig. 2
: Rear of the 817 Bioscan
4
817 Bioscan
4
1.2 Parts and controls
Mounting screw
4
for fastening the rear panels 5 / 6
12 Lower connections for remote lines
(not used with IC Net)
Detachable rear panel
5
access to top part of the inner compartment
6 Detachable rear panel
access to bottom part of the inner
compartment
7 Analog signal (REC.)
AD/DA-converted signal
(not used with IC Net; see chapter 6.1
for details)
8 Analog signal (Int.)
Original signal
(not used with IC Net; see chapter 6.1
for details)
9 Manual valve
(not used with IC Net)
10 Model plate
with technical data serial number
13 Outlet for spilled liquid
for discharge of spilled liquid from the
inner compartment
14 RS232 interface
connection of the PC; see chapter 6.1
for details
15 Upper connections for remote lines
Aux 1, Aux 2 as well as 1 and 2 are
only
used with IC Net
16 Fuse holder
changing the fuses, see section 2.2.1
17 Mains connection plug
mains connection, see section 2.2
18 Mains switch
to switch instrument on and off:
I = ON 0 = OFF
11 Electric valve
(not used with IC Net)
817 Bioscan
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1 Introduction
19
20
21
34
22
23
24
25
26
27
33
1
Metrosep
Carb 1
32
2
29 31
28
: Interior of the 817 Bioscan
Fig. 3
30
(with permanently attached accessories)
3
817 Bioscan
6
1.2 Parts and controls
Electrode cable (red)
19
connection for working electrode
27 Connection for elektrode cable
connection '1'
Electrode cable (blue)
20
connection for auxiliary electrode
28 Connection capillary
connection 709 IC Pump – pulsation
absorber
21 Column connection capillary
PEEK capillary
29 Connection capillary
connection pulsation absorber –
injection valve
22 Measuring cell holder 30 Pulsation absorber
6.2620.150
23 Au - Flow cell
6.1254.010
24 Electrode cable (black)
connection for reference electrode
25 IC column
e.g. Metrosep Carb 1 (6.1013.000)
31 Preheating capillary
6.1836.010, length: 3 m
32 Oven
33 Oven heating fan
for carbohydrate analysis
26 Column holder
34 Connection capillary
connection flow cell – waste
817 Bioscan
7
1 Introduction
1.3 Information about the Instructions for Use
Please read through these Instructions for Use carefully before you put
the 817 Bioscan into operation. The Instructions for Use contain
information and warnings to which the user must pay attention in order
to assure safe operation of the instruments.
1.3.1 Organization
These Instructions for Use 8.817.1003 for the 817 Bioscan provide a
comprehensive overview of the startup procedure, operation, fault rectification and technical specifications of these instruments. The Instructions for Use are organized as follows:
Section 1 Introduction
General description of instruments, parts and controls
and safety notes
Section 2 Installation
Setting up, mains connection, attachment of accessories, connection to IC system
Section 3 Operation
Detailed description of the operation
Section 4 Basics
Information about the pulsed amperometric detection
Section 5 Notes – Maintenance – Faults
Notes on ion chromatography, maintenance, fault rectification, diagnostic tests, validation
Section 6 Appendix
Technical data, standard equipment, options, warranty,
declarations of conformity, index
To find the required information on the instrument, you will find it an advantage to use either the Table of contents or the Index at the back.
As a supplement to the Instructions for Use, the 8.732.2003 Metrohm
Monograph "Ion chromatography" is also supplied. This provides an
introduction to the theoretical fundamentals and general information on
separating columns and sample pretreatment.
The 8.792.5003 Metrohm Monograph "Practical Ion Chromato-
graphy" is a practical textbook, which provides an illustrative introduction to the basic principles of ion chromatography and also describes
22 experiments covering the whole world of ion chromatography.
IC Application Notes for different applications of Metrohm IC systems
are also supplied.
8
817 Bioscan
1.3 Information about the Instructions for Use
1.3.2 Notation and pictograms
The following notations and pictograms (symbols) are used in these Instructions for Use:
<PARAM> Key
'Range' Parameter or entry value
35 Part or control of 817
22 Part or control of 709
Hazard
This symbol draws attention to a
possible danger to life or of injury if
the associated directions are not
followed correctly.
Warning
This symbol draws attention to
possible damage to instruments or
instrument parts if the associated
directions are not followed correctly.
Caution
This symbol marks important
information. First read the associated directions before you continue.
Comment
This symbol marks additional
information and tips.
817 Bioscan
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1 Introduction
1.4 Safety notes
1.4.1 Electrical safety
While electrical safety in the handling of the 817 Bioscan is assured in
the context of the specifications EN61010-1 / IEC61010-1, the following
points should be noted:
• Mains connection
Setting of the mains voltage, checking the mains fuse and the
mains connection must be effected in accordance with the instruc-
tions in section 2.4.
• Opening the 817 Bioscan
If the 817 Bioscan is connected to the power supply, the instrument
must not be opened (apart from the front door) nor must parts be
removed from it, otherwise there is a danger of coming into contact
with components which are live. Hence, always disconnect the
instrument from all voltage sources before you open it and ensure that
the mains cable is disconnected from mains connection 17 !
• Protection against static charges
Electronic components are sensitive to static charging and can be
destroyed by discharges. Before you touch any of the components
inside the 817 Bioscan, you should earth yourself and any tools you
are using by touching an earthed object (e.g. housing of the instrument or a radiator) to eliminate any static charges which exist.
1.4.2 General precautionary rules
• Handling of solvents
Check all lines of the IC system periodically for possible leaks. Follow
the relevant instructions regarding the handling of flammable and/or
toxic solvents and their disposal.
• Never block drain opening for spilled liquids
On the bottom of the interior directly below the front door there is a
drain opening 1. Spilled liquids can flow directly to the outlet 13 on
the back. Due to safety consideration, take care that these openings
may never be blocked.
10
817 Bioscan
2.1 Setting up the instrument
2 Installation
The 817 Bioscan is an electrochemical detector and can be included in
a modular IC system in two different ways:
1) The 817 Bioscan as the only detector in an IC system.
Apart from the 709 IC Pump, you also require an 812 Valve Unit with
one injection valve. In this MIC-8 modular IC system the 817 Bioscan is
intended for the analysis of sugars only.
2) The 817 Bioscan is included in an IC system as the second
detector. An example of this is given in Section 2.5 where an IC system
(MIC-9) is described for the simultaneous determination of sugars and
anions with chemical suppression.
These modular IC system configurations (MIC-1, MIC-2, etc.) are possible instrument combinations that have been properly tested. Under
www.metrohm.com
updated.
you will find further examples; these are continually
Following the general information about the installation of the
instruments, Sections 2.4 and 2.5 describe the two configurations
mentioned above including their control by the «IC Net 2.1» software.
2.1 Setting up the instrument
2.1.1 Packaging
The 817 Bioscan is supplied together with the separately packed
accessories in special packaging containing shock-absorbing foam
linings designed to provide excellent protection. The actual instruments
are packed in an evacuated polyethylene bag to prevent the ingress of
dust. Please store all this special packaging as only it can assure
transport of the instruments free from damage.
2.1.2 Check
After receipt, immediately check whether the shipment is complete and
has arrived without damage (compare with delivery note and list of
accessories in section 6.2). In the case of transport damage, see
instructions in section 6.3.1 "Warranty".
817 Bioscan
11
2 Installation
2.1.3 Location
Position the instruments in the laboratory at a location convenient for
operation, free from vibrations and protected against a corrosive atmosphere and contamination by chemicals. The same applies to all
other components of the IC system.
To avoid disturbing temperature influences on the insulated column
compartment, the entire system including pump and eluent reservoir
must be protected against direct sunlight.
2.1.4 Arrangement of the instruments
If the 817 Bioscan is used as a single detector (see chapter 2.4), the
709 IC Pump, 812 Valve Unit and 817 Bioscan are best stacked on top
of one another in this order.
In a two-channel system (see chapter 2.5), the optimum arrangement
(1, 2 or 3 towers) depends on the laboratory space available.
In general, the IC pumps should be set up at the very bottom and the
IC detectors at the very top.
2.2 Mains connection
Follow the instructions below for connecting to the power supply. If
the instrument is operated with a mains voltage set wrongly and/or
wrong mains fuse, there is a danger of fire!
2.2.1 Setting the mains voltage
Before switching on the 817 Bioscan for the first time, check that the
mains voltage set on the instrument (see Fig. 4) matches the local
mains voltage. If this is not
on the instrument as follows:
1 Disconnect mains cable
Disconnect mains cable from mains connection plug 17.
the case, you must reset the mains voltage
2 Remove fuse holder
Using a screwdriver, loosen fuse holder 16 below the mains
connection plug and take out completely.
3 Check and change fuse if necessary
Carefully take the fuses installed for the desired mains voltage
out of fuse holder and check their specifications:
100…120 V 3.15 A (slow-blow) Metrohm No. U.600.0021
220…240 V 1.6 A (slow-blow) Metrohm No. U.600.0018
817 Bioscan
12
2.2 Mains connection
V
V
4 Insert fuse
Change fuses if necessary and reinsert in fuse holder.
5 Install fuse holder
Depending on the necessary mains voltage, insert fuse holder in
the 817 Bioscan so that the corresponding mains voltage range
can be read normally and the adjacent white arrow points to the
white bar imprinted below the fuse holder (see Fig. 4).
2.2.2 Fuses
18
17
100 – 120
220 – 240
18 Mains switch
17 Mains connec-
tion plug
16
220 - 240 V
100 - 120 V
100 - 120 V
220 - 240 V
16 Fuse holder
Fig. 4
: Setting the mains voltage
Two fuses 3.15 A/slow-blow for 100…120 V or 1.6 A/slow-blow for
220…240 V are installed in fuse holder 16 of the 817 Bioscan as standard.
Ensure that the instrument is never put into operation with fuses of a
different type as this could cause a fire!
817 Bioscan
For checking or changing fuses, proceed as described in section 2.2.1.
13
2 Installation
2.2.3 Mains cable and mains connection
Mains cable
The instrument is supplied with one of three mains cables
• 6.2122.020 with plug SEV 12 (Switzerland, …)
• 6.2122.040 with plug CEE(7), VII (Germany, …)
• 6.2133.070 with plug NEMA 5-15 (USA, …)
which are three-cored and fitted with a plug with an earthing pin. If a different plug has to be fitted, the yellow/green lead (IEC standard) must
be connected to protective earth (protection class 1).
Any break in the earthing inside or outside the instrument can make it
a hazard!
Mains connection
Plug the mains cable into mains connection plug 17 of the 817 Bioscan
(see Fig. 2).
2.2.4 On/off switching of the instruments
The 817 Bioscan is switched on and off using mains switch 18. Do not
switch the IC components on before all cable connections have been
established.
817 Bioscan
14
2.3 Capillary connections
2.3 Capillary connections
The capillary connections described below are necessary for operating
the 817 Bioscan in a simple IC system for carbohydrate analysis (MIC-
8). The electronic connections and control by IC Net 2.1 are described
in Section 2.4.
Section 2.5 contains the description of a combination of this IC system
with an IC system for the determination of anions with chemical suppression (MIC-9). The necessary changes to the capillary connections
are described there.
2.3.1 Connection of the 6.5324.000 Bottle rack (option)
The optionally available 6.5324.000 Bottle rack for supply vessels can
be placed on top of the 817 Bioscan. The tubing connections to the 709
IC Pump are described in the corresponding leaflet.
2.3.2 Connecting the pulsation absorber
To protect the column material from any pressure shocks which the injection may cause we recommend the insertion of the 6.2620.150 MF
Pulsation absorber between the high-pressure pump and the injection valve. This is done as follows (see Fig. 3):
1 Connect the pulsation absorber
• Place pulsation absorber 30 on the floor in the interior of the
817 Bioscan.
2 Connection to pump
• Connect the 6.2821.100 PEEK Filter unit described in
Section 2.3.5 of the 8.709.1021 ‘Instructions for Use’ to connection 23
• Lead PEEK capillary 28 or 43
the filter unit outwards through opening 2 of the Bioscan and
attach it to one of the 52 connections on the top side of the
pulsation absorber.
3 Connection to injection valve
• Connect PEEK capillary 29 to the second connection 30 of
the pulsation absorber, lead it out through 2 of the Bioscan
and connection it to the injection valve on the 812 Valve Unit
(connection 17).
The pulsation absorber is filled with isopropanol and must be rinsed
with eluent before a separating column is connected (see Section
2.3.7).
of the 709 Pump by using PEEK capillary 22.
from connection piece 47 of
817 Bioscan
The 6.2620.150 Pulsation absorber can be operated in both directions.
15
2 Installation
2.3.3 Connection of the injection valve
Connect all capillaries to the injection valve according to the following
description:
46 45
45
36
21
4447
Fig. 5
: Connections at the injection valve of 2.812.0010 valve unit
Connection for syringe tubing
42
PTFE tubing with coupling
(6.2744.120) and syringe (6.2816.020,
without needle)
43
Connection for aspirating tubing
PTFE tubing for aspirating sample
44 Connection for sample loop
20 µL PEEK sample loop (6.1825.210)
2.3.4 Mounting the column
The IC separating column (e.g. Metrosep Carb 1, 6.1013.000) is first
mounted in the 817 Bioscan as follows (see Fig. 3):
42 43
45 Eluent outlet
Connection to preheating capillary 31
(PEEK, 6.1836.010)
46 Eluent inlet
PEEK capillary connection 29 to
pulsation absorber
47 Connection for sample loop
20 µL PEEK sample loop (6.1825.210)
1 Connect column to the preheating capillary
• Remove the caps from column 25.
• Screw eluent outlet of preheating capillary 31 on to the inlet
end of the column (observe flow direction) .
2 Connect column to flow-through measuring cell
• Screw outlet end of column 25 on to eluent inlet 41 of the
flow-through cell using the 6.2744.130 KELF pressure screw.
3 Fix column
• Fix the column in column holder 26.
817 Bioscan
16
2.3 Capillary connections
2.3.5 Connection of the flowcell
The Flowcell (6.1254.010) is a part of the standard equipment of the
817 Bioscan. It is already mounted and ready to use.
37
36
38
35
39
41
40
: Connections at flowcell
Fig. 6
Check marks
35
For correct orientation of the top and
bottom part of the flow-cell
36
Mounting screws (4x)
37 Outlet for eluent
Connection of PEEK capillary by
6.2744.130 KELF pressure screw .
PEEK pressure screws may damage
the threads of the cell.
38 Working electrode connection
for red electrode cable 19
39 Auxiliary electrode connection
for blue electrode cable 20
40 Reference electrode connection
for black electrode cable 24
41 Inlet for eluent
Connection of PEEK capillary by
6.2744.130 KELF pressure screw.
PEEK pressure screws may damage
the threads of the cell.
817 Bioscan
1 Before starting up check all screws 36
• Use the Allen key provided to loosen all four of the 36 screws
by one rotation.
• Then slightly tighten the screws again, e.g. by holding the
Allen key between the thumb and index finger only.
• Finally tighten all four screws by a quarter rotation.
• Always tighten opposing screws, i.e. tighten one screw and
then the opposite screw.
• Avoid overtightening the screws as this could damage the
6.1254.020 Distance piece!
17
2 Installation
2 Connect the electrode cable
• The included 6.2156.000 Cable for connecting the measur-
ing cell is attached to connections 38, 39 and 40 of the
measuring cell using the three connectors 19, 20 and 24
(see Fig. 6).
• The other end of the cable is inserted into connection '1' of
the 817 Bioscan 817 with connector 27 (see Fig 3).
3 Connect the column
• Screw column connection capillary 21 of the column on to
eluent inlet 41 of the flow-through measuring cell using the
6.2744.130 Pressure screw.
4 Connect the waste container
• In order to ensure an adequate backpressure, screw a PEEK
capillary (approx. 1 m) on to eluent outlet 37 of the measuring
cell.
• Lead the end through opening 3 into a sufficiently large waste
container and attach it there.
5 Mount the measuring cell
• Attach the flow-through measuring cell to the holder 22 817
Bioscan which is provided for it (see Fig. 3).
• Rotate the holder and the cell so that the cell outlet is located
as high as possible. In this way it is easier for any air bubbles
that may occur to escape from the cell.
Never switch on the flow-through cell when this is
(a) not being rinsed through with a conducting eluent at the
time,
(b) not completely connected up, or
(c) when its outside is moist so that so a short circuit could
occur between the connections of working electrode 38
and auxiliary electrode 39!
2.3.6 Connecting the waste tubing
The 817 Bioscan has an outlet 13 on its rear panel for discharged
liquids to which the waste tubing can be attached. Proceed as follows:
1 Connect the waste tubing
• Attach matching tubing (e.g. Silicone tubing, optional) into
outlet 13 (see Fig. 2).
2 Lead the waste tubing to a drain
• Place the other end of the waste tubing in a drain and attach
it there.
817 Bioscan
18
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