Instrumental operation: ICS 3000 - NMBU

Instrumental operation: ICS 3000

Standard Operational Procedure

Jagger


Scope

The scope of this document is to present an overview of the Dionex Ion Chromatography System (ICS)

3000 and to provide the reader with guidelines for standard operations, routine maintenance and

simple trouble shooting. Detailed instrument description can be found in Dionex Reference Library (CD-

ROM) which contains all relevant and specific manuals.

Instrument overview

The Dionex ICS 3000 system is designed for ion chromatography (IC). It is compatible with high salt and

alkaline solvents and therefore fittings, tubings and interior of the system is made of PEEK (polyether

ether ketone) to prevent corrosion. It consists of a dual pump system with four channels on each pump.

It is capable of operating two columns with two different detectors at the same time and is equipped

with a pulsed amperometry detector (PAD) and a conductivity detector. In practice, the common use

currently will be one column at a time and primarily on Pump 1 and the PAD detector which will be

described here. The PAD is based on electrochemical detection and is in general superior for detection

of carbohydrates that are traditionally difficult to detect. The system also includes an eluent generator,

which can be used when ultra pure eluents are needed. In practice, the eluent generator has limitations

that make it difficult to use for many applications and it will therefore be bypassed for most

applications. The principle of the conductivity detector is that changes in conductivity is measured and

interpreted as analytes. This requires the installation of an ion suppressor prior to the conductivity

detector, which will suppress the inherently high conductivity coming from the eluents. Figure 1 shows

an instrument overview. As opposed to many other HPLC systems, the system should be operated with

more or less constant flow to avoid detrimental salt precipitations in the pump, column and detector. If

the flow will be stopped for a prolonged time period make sure to replace salt containing eluents in all

lines with e.g. 50% methanol (also to avoid microbial growth).

HPLC systems are delicate equipment and should be handled accordingly. Regularly check the status of

the instrument, preferably on a daily basis and always be careful when operating. Eluents should

ALWAYS be prepared from fresh top quality water (Milli-Q/18.2 MΩ·cm) and preferably sample

preparation should also be made based on this water quality.

The chromatographic system is controlled by a computer installed with Chromeleon™ 7 software

(hereafter CM) version 7.1.1.1127 for instrument control and data handling. All commands and

instrument handlings goes through this software.



Jagger

The system consists of 5 compartments/modules:

1. Autosampler

2. Dual pump

3. Eluent generator

4. Column compartment (lower part)

5. Detector compartment (upper part)

Figure 1: General system appearance of the ICS-3000



Jagger

Figure 2: Instrument Controller. Software unit that controls communication between computer and HPLC. Must be running

at all times when system is operated.

Communication between the computer (CM) and the HPLC is controlled by the Instrument Controller

Software found under the Start Menu/Programs/Chromeleon/Service Manager. Alternatively, double

click the Chromeleon Icon in the Windows task bar to open the Instrument Controller (figure 2)or right

click and select start Chromeleon Instrument Controller. When the HPLC is to be controlled/operated

the Instrument Controller must be running. Starting and stopping is done by pressing the command

“Start/Stop Instrument Controller” (figure 2). The window can be closed afterwards. The Instrument

Controller does not start automatically after a computer restart and it should therefore be checked that

it is running, before starting operation.

In the operating software (CM) each module has its own operational screen, which may be found as tabs

in the top of the window (figure 3).

Window for starting and stopping

Instrument Controller. Can be closed

after use.

Icon informing about Instrument

Controller status. Green: Controller

running. White with red cross:

Controller stopped.



Jagger

Figure 3: Module and system overview from controlling software Chromeleon™ 7. Unmarked tabs: “DC”; Detector

compartment. “EGC1”; Eluent generator. “Audit”; Audit trail logging every event in the system. “Startup”; Programmed

system startup (not recommended to use unless you know what you’re doing).

Operating the pump

The internals of the dual pump system is depicted in figure 4. The lower half of the compartment is

Pump 1 and the upper half is Pump 2. The pumps are operated at flow rates between 0 and 10ml/min

and max pressure is 5000 psi. Each pump is connected to four eluent channels (A, B, C and D). A Rear

Seal Wash system is installed to periodically flush the backside of the pump heads in order to avoid salt

precipitations and corrosion. Wash setting may be “Interval”, “Automatic” or “Off”. Correct setting is

“Interval”, but the rear seal wash solutions needs to be changed on a weekly basis and consists of 20%

methanol/ethanol (figure 4). Note dates of change in the equipment logbook.

Switching between

Instrument and Data panel

Overview Autosampler PAD detector Queue Pump

System 1 – PAD

electrochemical

detection



Jagger

Figure 4: Internals of Dual Pump. Pump 2 is placed in the upper half of the compartment, Pump 1 in the lower half. Note the

screw valve at position 3 is the purge valve on Pump 2. Equivalent purge valve on Pump 1 found in the same position.

Position 11 is the rear seal wash reservoir which should be changed on a weekly basis (20% methanol or ethanol).

Preparing eluents

The eluents for ion chromatography are crucial for good performance and the chromatography (column)

is extremely sensitive to contaminations in the eluents. It is therefore of outmost importance that extra

care is taken when eluents are prepared. Remember that the lifetime of eluents for ion chromatography

is also very short compared to other types of chromatography. The biggest risk of contamination is

dissolution of carbonate in the eluents and this occurs when eluents are exposed to atmospheric air.

This carbonate accumulates on the columns stationary phase and reduces separation capacity and

carbonate has a higher push effect as mobile phase then the others regularly applied. It is impossible to

completely avoid carbonate contamination but with the proper precautions it can be minimized and

allow a certain lifetime of the eluents (max. 48 hours for top performance). This includes that



Jagger

headspace in the eluent bottles must be saturated with nitrogen to minimize the amount of carbon

dioxide. Nitrogen is delivered by a nitrogen generator via a restriction valve and distributed between

three of the four channels (channel D does not get nitrogen supply). Three different eluents are made to

accommodate the range of analytical methods and all three are changed every time new eluents are

prepared. The eluents are always prepared in the same way and always placed on the following

channels:

Channel A: 0.1 M NaOH

Channel B: 1 M NaOAc in 0.1 M NaOH

Channel C: Milli-Q water

Eluent A and B are most labile for carbonate contamination and should be handled accordingly.The

following is a description for generation of each eluent but should be performed alongside so that all

three eluents are done approximately at the same time. All eluents are sensitive to atmospheric air.

Therefore – don’t leave any of the newly prepared eluents standing without N2 over pressure in the

headspace for more than 15 minutes.

Before any eluents are made, turn off the flow on the system and release the nitrogen pressure on the

restriction valve positioned on the backside of the autosampler. Empty all eluent bottles and rinse them

thoroughly in Milli-Q water. Special pressure stable eluent bottles are used since strong corrosive

eluents are pressurized during running. Therefore, don’t send the bottles off to washing, but rinse them

properly before reuse.

Eluent A: 0.1 M NaOH (2L)

1. Measure out 2 L of Milli-Q water in a measuring flask. Make sure to use clean glassware.

2. Poor the two liter of water into the eluent bottle named Eluent A (0.1 M NaOH).

3. Loosely place a lid on the bottle, but don’t tighten it.

4. Place the eluent bottle in the sonication bath and adjust the water volume in the bath to the

level indicated as working level. The sonication bath will fit two eluent bottles at a time.

5. Sonication for 20 min. DON’T put your hands into the sonicator while it is operating and close

the door to the room when you leave it.

6. When the sonication is done, close the lid securely and move the eluent bottle to the HPLC.

7. Connect channel A to the eluent bottle and start the nitrogen flow. Exchange the air in the head

space by repeatedly opening and closing the lid on the bottle to relieve pressure. A total of three

reliefs are sufficient.

8. Use a sterile plastic pipette with a total volume of 10 ml and a manual “pipette bulb” for

pipetting a concentrated 50% NaOH solution (Fluka, 71868-1L). The solution is stored in the

exhausted waste cupboard underneath the HPLC and is extremely concentrated. ALWAYS wear



Jagger

proper personal protection equipment (glasses, gloves and lab coat/acid rubber coat) when

handling this solution. I am not kidding – getting that stuff in your eyes will ruin your eyesight!

9. Check the level of NaOH solution in the bottle – if the content is less than half of the original

replace with new to prevent risk of carbonate contamination in the NaOH solution. Place the

remainder of the solution in the toxic cupboard and mark it with “COMMON USE” for other

purposes than HPLC.

10. Pipette 10.4 mL to the 2 L bottle of eluent. Be careful when pipetting, as the solution is very

viscous. Don’t dip the entire pipette into the solution – only the tip underneath the surface.

Close the lid to the NaOH solution immediately after use and only use the same pipette ONCE in

the solution. Don’t double dip – take a new pipette.

11. Dispense the NaOH solution to the water in the eluent bottle underneath the water surface and

be patient as the liquid will flow very slowly (viscous). Slowly rinse the inside of the pipette by

gently pipetting up and down 3 times.

12. Close the lid to the eluent bottle and exchange the headspace air with nitrogen as done in step

7. Swirl the eluent bottle carefully – DON’T shake it, to distribute NaOH evenly.

Eluent B: 1 M NaOAce in 0.1 M NaOH (1L)

1. Weigh 82.04 g anhydrous sodium acetate (Sigma-Aldrich, 71183-1KG,or Fluka). It is important

that the correct quality of sodium acetate is used.

2. Poor this into a 1 L measuring flask (use clean glass ware) and add about 0.8L Milli-Q water.

3. Add a magnetic stir bar and place the solution on a magnetic stirrer till you have a complete

dissolution.

4. Remove the magnetic stir bar (without dipping the magic stick into the solution!) and fill the

remaining volume to 1 L (with freshly tapped Milli-Q water).

5. Mount a 0.22 µm sterile filter cup on top of the cleaned eluent bottle (f.ex. 0.22 µm stericup GP

Millipore Express PLUS membrane, 250 ml funnel, 45 mm neck size). Apply vacuum to filter the

eluent.

6. Loosely place a lid on top of the eluent bottle. Don’t tighten it.

7. Place the eluent bottle in the sonication bath, make sure water surface in the bath is at the

working level.

8. Sonicatefor 20 min. DON’T put your hands into the sonicator while it is operating and close the

door to the room when you leave it.


10. Connect channel B to the eluent bottle and start the nitrogen flow. Exchange the air in the head





Jagger

11. Follow the procedures for NaOH addition as described for Eluent A step 8-12. EXCEPT! Only add

5.2 mL NaOH to the 1 L NaOAc-solution (ie. there should be the same NaOH concentration in

both channel A and B).

Eluent C: Milli-Q water

1. Fill the rinsed eluent bottle with Milli-Q water freshly tapped from the system.

2. Loosely place a lid on top of the eluent bottle. Don’t tighten it.

3. Place the eluent bottle in the sonication bath and adjust the water volume in the bath to the

level indicated as working level.

4. Sonicatefor 20 min . DON’T put your hands into the sonicator while it is operating and close the

door to the room when you leave it.


6. Connect channel C to the eluent bottle and start the nitrogen flow. Exchange the air in the head



End the preparation procedure by purging all three channels as described below and note in the logbook

date of eluent change.

Purging eluents

When eluents are changed the pump needs to be purged to remove air bubble formation and old eluent

from the tubes. The system is sensitive to air (as all HPLCs) and purging is therefore important. Purging

the eluent channels is done be applying high flow through the specified channels and will create high

back pressure on the system, if the purge valve has not been open (Open the purge valve (figure 4) by

approx. half a turn). This will direct the flow path to a waste tube and relieve the pressure on the system

(and the column in particular).

Once the purge valve has been opened choose the appropriate channels and flow distribution to purge

in CM and activate the purge/prime function (figure 5). It is possible to purge more than one channel at

a time by setting the percentage distribution between the channels as desired (equal distribution, 50:50

when two channels, 33% when three and 25% when all four channels are to be purged). The software is

preprogrammed to perform purging for 500 seconds at 5.7 mL/min. Repeat purging (if necessary) until

air bubbles are no longer visible in the eluent channels from the bottles to the pump.

IMPORTANT: Before purging the software will inform that this operation will generate a

high flow through the system and that the purge valve should be open before

executing. Press “Execute despite warning”, if the purge valve is open. If the system

senses any pressure increase (even minor) during purging, the command will be

terminated. Make sure that the valve has been sufficiently loosened and try again.



Jagger

Figure 5: Pump tab. Relevant functions marked for purging, flow settings ect.

Once the purging is done, close the purge valve again tightly to redirect the flow path to the column. If

the valve is not tightened enough, the pressure during operation will not be sufficiently high meaning

that the flow delivery to the column is insufficient. No warnings about low pressure will occur, unless

pressure is completely lost (less than 200 psi) so please do observe that correct pressure is obtained

during start up. Normal operating pressure for CarboPac PA1 incl. guard is at least 15-1700 psi (this

includes a pressure restrictor coil at 1000 psi).

Operating the autosampler

Samples are placed in the autosampler prior to operation. The front door of the autosampler is opened

(held in place by a relatively strong magnet – pull it hard!) and the vials placed in the sample tray (figure

6). On the outside of the autosampler in the lowest left corner is the main power switch – not a door

opener! Only press this button if you really want to cut the power to the autosampler. Sample position

in the tray is straight forward numbering from 1 to 100. Each position is marked on the tray. The tray

may be removed from the autosampler when samples are being loaded (be careful…). The sample tray is

placed in a cooled compartment and the autosampler will not operate if the front door is open. A sound

alarm will occur if the door is left open (turns off again when the door is closed). Don’t open the door if

the robot arm is moving as this will immediately interrupt the movements. The front panel will inform

Prime/purge

function

Pump motor

Flow settings

Pressure

indicator

Solvent

distribution

Module connection

Retention time count



Jagger

about the “state” of the autosampler: “Idle” means that the autosampler can be opened for vial

handling (this is also the case for idle status during a sequence run). Otherwise, the front panel is used

for manual handling and is not used during routine operation. All normal operational control is handled

via CM. Figure 8 shows the routine operations of the autosampler from CM. The sampler has a fixed

volume loop and if this needs to be changed, call a super user. Partial injection may be applied if

necessary, but will result in increased standard deviations, but please consult super user or manual. NB:

Be aware that the sampler consumes approx. 40 µL sample for a 2 µL injection.

On top of the autosampler (towards the back) is the syringe tower (figure 6). This is where sample,

buffer etc. is taken and measured before injection to the column. Make sure no air bubbles are trapped

in the syringe by priming sampler before you start your analysis. The syringe is connected to a wash

solution consisting of 20% methanol.

The sample is introduced via the injection port, via the diverter valve (position 1 directs sample to

system 1), via the loop (flow path is via the loop when the injection valve is in inject mode, meaning

always except during an injection cycle) to the column. When the needle leaves the inject port (figure 7)

the bed should be dry with no droplets/spillage. If not, something is obstructing the injection and

poor/no signals will be observed in the following chromatogram. The needle is washed before each

injection in the flush port (figure 7) next to the injection port. Here, wash solution will appear (and may

spill) which is normal.



Jagger

Figure 6: Autosampler with sample tray dismounted from the tray compartment

Figure 7: Injection and flush port in autosampler

Diverter valve



Jagger

Figure 8: Autosampler tab in CM.

Operating the column compartment

The column compartment is the module where the column is connected (figure 9). During operation the

front cover of the column compartment should preferably be closed and especially if temperature

control is applied (improved reproducibility is observed when temperature is set slightly above

ambient). As mentioned above the flow is directed from the autosampler via the left injection valve (for

pump 1) to the column. The loop is also mounted on the injection valve. Do not mess around with the

tube connections here! The column is fitted in an appropriate holder in the back of the compartment.

Prime syringe

Temperature control in sample tray

Autosampler status

Sample loop size

flush sample

Diverter valve/inject valve



Jagger

Figure 9: Internals of column and detector compartment. Blue arrow indicates flow direction on the column, which should fit

with column flow directions. Upper thrid of the compartment is not present in this particular instrument set up.

When a column is fitted and flow is applied please ensure that there are no leaks. Leaks at the column

inlet will result in pressure decrease, whereas leaks at the column outlet may not affect the pressure but

will affect the detector signal and potential flooding of compartment. If leaks are observed or registered

by the system, further tighten the connections either by hand or by a wrench key (be careful, PEEK

connections are weak).

Column temperature is specified in CM (figure 10) and the desired temperature is written in the field for

“Set”. Press Enter to execute the command. The field “Actual” will inform about the actual temperature

in the column compartment (which initially is not necessarily the same as the column temperature).

Analytical Column flow direction

Left injection valve for Pump Guard column – Note flow

direction: Left to right!

Analytical column – Note

flow direction: Right to left!



Jagger

Figure 10: Column compartment tab for controlling column temperature.

Operating the PAD detector

The detector is placed as can be seen in figure 9 (item 2) and a close-up photo in figure 11. The detector

consists of a gold working electrode and a pH reference electrode. The pH reference electrode (number

3 in figure 11) has very limited needs for maintenance during routine operations and should not be

removed from its socket. The working electrode on the other hand is disposable and needs to be

changed regularly. Routines for changing working electrode are described below. The detector can only

be turned on when flow is on the system and a pump link exists where the cell voltage on the electrode

is turned off, if the flow is stopped. However, cell voltage should only be turned on during sequence

running and other diagnostic runs, since prolonged voltage will decrease the electrodes life time. When

running, the electrode senses two signals, a total signal (a background signal that depends on the type

of eluents and state of the electrode) and a baseline/real sample signal which can be autozero’ed. The

detector responds to any substances that are either oxidizable or reducible and the electrical output

results from an electron flow caused by the chemical reaction that takes place at the surface of the

electrodes. The eluents are highly charged and this can be observed as the background (Total) signal.

Normal operating Total Signal should be between 15 and 35 nC. The reference electrode is capable of

Temperature control



Jagger

adjusting/compensating changes in background signal when gradient elution profiles are run and in that

sense maintain a stable baseline. If the reference electrode is not installed/out of order, the baseline will

rise considerably when gradient profiles are used. PAD is a particularly strong detection principle for

carbohydrates, because the highly alkaline eluents will charge the hydroxyl groups and therefore give

rise to signals in the detector. Furthermore, this detection principle is highly sensitive and therefore

makes it superior for carbohydrate analysis.

Figure 11: PAD electrode

From column

To waste



Jagger

Figure 12: PAD tab in CM.

Changing working electrode

When the Total Signal becomes too high (above 40-45 nC) or the signals during running behaves strange

(extremely high signal or seemingly no signal at all/increased background signals), changing the working

electrode may be needed (The electrode has a life time normally between 1 and 3 weeks continuous

operation). If you suspect the electrode may be worn out, check last date of change in the logbook to

verify. New users should consult a super user before changing the electrode the first time. Hereafter

do as follows:

1. Turn off the flow and thereby the cell voltage.

2. Disconnect the inlet and outlet of the detector (by the incoming and out coming of the red

tubing in figure 11). You may need a wrench key.

3. Unplug the blue and the yellow power plugs (figure 11).

4. Grab the electrode by two hands and pull it firmly towards you in one motion (yes, it will come

off).

5. While performing the following steps, try to keep the entire electrode in an upright position. The

pH reference electrode contains KCl which may leak if turned upside down.

6. Loosen the big knob (picture A, figure 13), press the two side-wings on the clips that holds the

spacer block in place and take off the entire clips.

Turning on/off cell voltage

Signal and Total Signal (background)

Autozero (of Signal)



Jagger

7. Remove the spacer block and now the individual parts of the electrode are visible (picture B,

figure 13). The working electrode is the disposable one but take EXTRA care of the gasket. They

are not supplied enough to be changed every time. Unless it is torn or otherwise damages it

should be reused.

8. Inspect the working electrode on the “gold”-side. Salt precipitations or loss of connection

between top and bottom is reason for loss of signal. Discard the electrode.

9. Inspect the cell body and carefully rinse it if visible salt precipitations are seen (picture B, figure

13).

10. Take a new working electrode from the package, but ONLY lift it in the snip of white tape that

goes outside the actual electrode which says “lift here”. Use a pair of tweezers if necessary.

11. Place the working electrode and the gasket back on the cell body. Note that the two positioning

holes in the gasket are different in shape, which compared to the spacer block indicates the

positioning. The same goes for the working electrode. Furthermore, the electrode should be

placed so that the golden parts face the cell body and so that the small circle fits over the slit in

the gasket. Do not bend the working electrode in any way! Position it precisely over the two

rods and slide it onto the cell body. See correct placement of the working electrode in picture C,

figure 13.

12. Place the spacer block and the clips back on the electrode and turn the big knob until you hear a

click. This is as tight as it gets.

13. Reposition the entire electrode in the detector compartment (it needs to say click), re-plug the

electrical cords (yellow in yellow, blue in black!), reconnect column and waste tubings, restart

flow on system (make sure there are no leaks!!) and set cell voltage on.

14. Monitor baseline for a while (approx. 20-30 min). The signal can be quite unstable in the

beginning (and very high), but will eventually stabilize. Total signal should preferably stabilize

between 15 and 30 nC.

Figure 13: Changing disposable

gold electrode

Working

electrode

Spacer block

Gasket

Cell body Spacer block

Spring- clips

A B C



Jagger

Waste collection

From the detector the flow stream is passed to a waste container standing in the exhaust cabin

underneath the system. This waste container needs to be checked regularly and emptied. The waste is

almost exclusively consisting alkaline salts and can therefore be poured in the drain. For the same

reason – spillage of waste will generate vast amounts of salt precipitations and should therefore be

avoided.

Starting up for operation

1. Make sure that eluent level is good and check the last date for eluent preparation. If the eluents

are more than 2 days old, consider if you should make new eluents before your run. If preparing

new eluents, remember to purge all channels in use (A,B and C) after eluent preparation

2. Check that the correct column is on.

3. Check that flow is on and pressure correct (column flow: 0.25 ml/min, pressure: 1500-1700 psi)

4. If column temperature control is applied make sure that the column compartment is closed

(lower door).

5. Turn on the detector cell (apply cell voltage).

6. Start “Baseline Monitoring” (see figure 14) and watch the detector signal stabilize.

7. If the signal does not stabilize or if the total signal is very high (above 45 nC) it may be time to

change the working electrode. Follow the procedure for doing so, if necessary.

8. Prime sampler, flush sample loop (make sure no air is entrapped in sample syringe)

9. Place the samples in the autosampler and prepare the sequence (see section below). If starting

up after eluent change, the first sample should ALWAYS be a blank sample running the

instrument method: “Start after eluent change”. This is to ensure regeneration of column after

possible carbonate contaminations. Afterwards condition the column with 1-2 blank runs with

the instrument method you intend to use before injection of the first real sample/standard.

These conditioning blanks should always be run no matter if eluent has been changed or not.

10. Stop baseline monitoring.

11. Start the run either by starting the sequence directly or by queuing the sequence (add to queue

in the “Queue” tab). Press “Ready Check” to get an estimate of eluent use during the entire

queue.



Jagger

Figure 14: Queue tab. Here sequences can be queued for automatic start and general checking of system status (ready check)

is performed.

Setting up a sequence in CM

The data structure in CM is shown in figure 15, which is initially divided between different instruments.

The PAD detector is set up to system ED_1. Hereafter, a general data folder contains a folder for each

user, specified by name. Maintain this order of data structure and save your sequence under your own

name.

Monitoring baseline.

Monitor pump pressure Queue tab. For

queuing several

sequences

Adding a sequence

to the queue.

Ready check. Testing

if the system is

ready for operation.

Gives an estimate of

eluent use.

Successful ready check



Jagger

Figure 15: Data structure in CM. Note the switch between Instrument and Data panel.

When making a sequence, choose “Create” from the top menu and choose “Sequence”. Hereafter a

wizard will start, that will guide you through setting up of a sequence:

1. Choose system: ED_1

2. Write a recognition name that it will be copied to for each sample (may be edited later), choose

number of samples, number of injections pr. sample (most often 1), position of 1st

vial and

injection volume (uL), normally the same as the sample loop size.

3. Choose Instrument Method (analytical run method) by accepting the lastly used method or by

browsing through folders. Choose Processing Method if applicable. Can also be added later. Set

default channel to ED_1.

4. Write a comment describing the particular samples (not obligatory).

5. Press finish and save the sequence to your folder under an appropriate name starting with the

following date format: YYMMDD_

Figure 16 shows and example of a prepared, finished sequence. Most fields in the sequence can be

edited before, during and after the run. This includes: Name, Type (blank, unknown, standard), Level

(only relevant for quantification), Position (vial position in the autosampler), Volume (injection

volume), Instrument Method (analytical run method, can only be edited before running) and

Switching between

Instrument and Data panel



Jagger

Processing Method (qualititaive/quantitative method). Some fields have scroll down menus, others

are text fields. Always press “Save” after editing any fields. A shortcut called “Fill Down” is made for

fast assigning sample position in the autosampler. Mark the “Position” field from the position of the

first vial, press F9 and choose “Renumber”. Press enter. Alternatively, select the Fill Down menu

(figure 17), choose Renumber and write the name of the first sample position. Press OK. Note that

the fill down function may be used in all the other columns as well.

Figure 16: Prepared, finished sequence.

IMPORTANT: Changing any of these fields mentioned above will not affect the

results, but changing the field “Status” will erase data behind, if the samples have

been run; “Idle” indicates a sample not yet analysed, “Finished” indicates a sample

analysed and therefore a file containing data. “Interrupted” is a sample stopped

during analysis due to some sort of error message, may also contain data, but

sometimes not useful data.



Jagger

Figure 17: Fill down menu for fast sample position assignment. Press “Fill Down” or F9 to open window.

Alarms and warming

Pressure alarm

If the pressure falls below or reaches the upper limits of the settings (which are mainly set to protect the

column) a pressure sound alarm will be initiated. At the same time, all flow will be stopped. High

pressure may occur if the samples contain insoluble particulate material, if the solvent concentration

suddenly changes, temperature control fails or if the column is overloaded with sample due to many

injections without regeneration. Loss of pressure may occur if the eluent bottles are empty, air is

introduced in the system or if leaks happen on the “upstream”/pressure side of the column. If leakage is

the cause, the system will detect the compartment where this is occurring via leak detectors and the

audit trail will inform about this.

Identify the source of the pressure violations, take proper action and restart the system.

Leak/humidity alarm

Identify the reason for leakage (which compartment) and correct if possible. Dry up any spillage in the

compartments with soft tissue paper.

Fill Down menu

Marked cell for

1st

vial position



Jagger

No pressure?

Check that the purge valve is properly closed. Check that all fittings are securely tightened. If this is not

enough purge the channels as air may have been trapped in the pump.

No connection to modules?

Check that the Instrument Controller is running (figure 2). When this is positivly running, reconnect the

non-connected modules by pressing “Connect” in each module tab (see figure 5 or 8). If there is still no

connection restart the computer and start the Instrument Controller again. Still no connection – call

security.

Software error messages

Close all active windows and restart CM. Still software errors, restart computer and start the Instrument

Controller again. Still errors – call security.

No contact with Autosampler

The autosampler needs a restart once in a while. If no contact can be achieved through the software,

turn off the autosampler (power button lower left corner, front). Leave it off for 1-2 minutes and start

again. It will now perform a few self tests (this takes a couple of minutes. When this is done, reconnect

in CM. If there is still no connection, call security.

Call security

For more advanced guidance either contact Super users or consult the general equipment manuals

found on the CD-rom: ”Reference Library” provided by Dionex.

Super users:

Jane Wittrup Agger ([email protected]; 6496 5856)

Bjørge Westereng ([email protected]; 6496 5909)

Documents

Instrumental operation: ICS 3000 - NMBU