CCC Instruments

Science & Solutions

NOTE.

Please check the instruction manuals your system was supplied with as some systems may differ from standard configurations.

 

The instruments covered by this section are the Spectrum and Midi.  Potential faults:

  1. Flying leads.  Flying leads are a consumable item and need to be replaced at regular intervals.  If flying leads are not replaced regularly, fitted as described in the appropriate operating manual and maintained as described in the operating manual the flying leads will develop leaks.  To stop leaks from flying leads a programme of preventative maintenance needs to be developed in accordance with the standard practises of the user’s organisation based upon the information provided in the operating manual.  However, the following tips will help determine the potential cause of a leak:
    1. On the Spectrum check that the loops of flying lead tubing on the bobbins have not become too large and extend beyond the outer edge of the bobbin.  These loops should as shown in figures 6 and 7 in the Spectrum operating manual.
    2. Check that the fittings where the flying leads are connected to the bobbin are tight enough to create a seal.
    3. If the above steps do not uncover the cause of the leak remove the flying leads and then examine the leads to find the leak.  Once the leak has been found please photograph the damage and send the picture to DE (info@dynamicextractions.com) and then determine the following:

i.The distance between the leak and the end of the tube connected to the bobbin.  Provide DE with this measured dimension to help determine the cause of the failure.

ii.Whether the leak: is a split in the tubing or a flatten section of wear through the side of the tubing or appears to as a short cut along the length of the tubing.  Provide DE with a description of the type of leak to help determine the cause of the failure.

  1. With the information provided in the previous step DE will be able to advise upon the cause of the failure and a solution to prevent similar failures in the future and prolong the service life of flying leads.
  1. Instrument stops spinning and a red LED is flashing on the front of the instrument.  This shut down is has been caused by either an imbalance or the instrument becoming too warm.  To determine the type of failure switch the “empty-off-run” switch to the “off” position and note the temperature of the instrument.  If the temperature is greater than 45°C then an over temperature shut down has occurred.  This is confirmed by opening and closing the instrument door and the red LED continuing to flash.  If the LED continues to flash go to “Over-temperature shut down” and if the LED stops flashing go to “Imbalance shut down”.
  2. Imbalance shut down.  To remove an imbalance follow steps a to h if no valve box is fitted to the instrument and steps i to n when a valve box is fitted.
  3. Check that the flying leads are correctly labelled for the ends of the columns to which each is connected.
  4. Ensure that the columns are connected as follows: PC2 to PP1, AC2 to AP1 and then AC1 to PP2.
  5. Connect the pump to AP2 and place outlet PC1 back into the water container from which the pump is taking water.  Note the level of the water in the container.  
  6. Turn the speed control knob fully anti-clockwise.  Switch on the rotation in the “Run” (clockwise) direction and start pumping the water into the desired column at 5ml/min for a time equal to the combined column volumes divided by 5 (ml/min).
  7. This will expel any remaining air from the columns.  Now determine if the level of the water is lower in the container than at the start.
  8. Then slowly increase the rotational speed to operational speed (2100 rpm for Mini, 1600 rpm for Spectrum and 1400 rpm for Midi) over a period of 10 minutes, the imbalance should now not occur.
  9. If the instrument shuts down due to an imbalance turn the speed control knob 1 full turn anti-clockwise cancel the imbalance alarm by opening and closing the instrument door and the switch on the clockwise rotation again.  Then continue increasing the rotational speed to the operating speed again over a period of 5 minutes.
  10. Check that the flying leads are correctly labelled for the ends of the columns to which each is connected.
  11. If this does not remove the imbalance contact DE and ask for Technical Support.
  12. Ensure that the connections between the valve box and the instrument are as described in the instrument and valve box operation manuals.
  13. Switching the NP/RP valve to NP and turning the speed control knob fully anti-clockwise.  Switch on the rotation in the “Run” (clockwise) direction and start pumping the distilled water into the desired column (5ml/min for the analytical column, up to 25ml/min for the semi-preparative Spectrum column and up to 100ml/min for the Midi preparative column).
  14. When a volume of solvent equal to the column volume has been pumped into the column slowly increase the rotational speed to the operating speed over a period of 5 minutes.
  15. If the instrument shuts down due to an imbalance turn the speed control knob 1 full turn anti-clockwise cancel the imbalance alarm by opening and closing the instrument door and the switch on the clockwise rotation again.  Then continue increasing the rotational speed to the operating speed again over a period of 5 minutes.
  16. If this does not remove the imbalance contact DE and ask for Technical Support.
  17. Check that the chiller is switch on and that the chiller’s set temperature is adjusted to the usual temperature for operating the HPCCC instrument.
  18. Check that the chiller is connected to the HPCCC instrument and that there are no coolant leaks.  If there are coolant leaks contact DE for further assistance and replacement parts.
  19. Check that the coolant within the chiller is free of algae and is circulating through the HPCCC instrument.
  20. Check that temperature control on the HPCCC instrument is switched on.
  21. One the Mini and Spectrum instruments open the door of the instrument and check the cooling fan is sucking air in beneath the rotor and blowing air out above the rotor.  If the fan is not operation contact DE for further instructions.
  22. Now reduce the set temperature of HPCCC instrument to 0°C and check the speed of the cooling fan and the flow of air through the fan increases significantly.  If there is no increase in fan speed contact DE for further advice.
  23. Do not switch off the power to the instrument.
  24. Unscrew the 2-off retaining screws in the right hand side panel and then carefully lean the top edge of the side panel away for the instrument.
  25. The error code is then displayed in 3 red letters within the right hand end of the instrument.
  26. Note the error code displayed and send the code to DE via info@dynamicectractions.com  DE will use the code to determine the cause of the shut down.
  27. Reposition and secure the right hand side panel in place.
  28. Errors codes in the drive system are cleared by switching the instrument off for 30 seconds and then switching back on.  However rotation of the instrument should not be started again until you have been contacted by a representation form DE.
  29. Open the door of the instrument and determine if the bobbin(s) has axial movement of approximately 0.5 to 1mm.  For the Mini and Spectrum instruments hold the rotor still using one hand and using your other hand try to move a bobbin towards the back and front of the instrument.  For the Midi hold the rotor still and try to move a bobbin to the left and right.  If a bobbin cannot be moved or is difficult to move it can be an indication that there is a problem with a bobbin bearing.
  30. Switch off the electrical power to the instrument, remove the mains lead and alarm cable if fitted and remove the right hand side panel.  This panel is removed by unscrewing the screws that attach the panel to the case.
  31. Reconnect the mains lead and switch the electrical power back on.  The drive inverter has a red LED display on its front.  Take great care to prevent electrical shocks press the white dial on the front of the inverter and turn the dial in either direction until “SUP” appears and then press the dial.  Turn the dial again until “OPr” appears and press the dial again.  Older versions of the drive inverter use a up & down arrows and a “ENT” button.
  32. Now switch on the rotation of the instrument and set the rotational speed to maximum.  Set instrument temperature to 30°C and allow the instrument to reach this temperature and then record the reading on the inverter and rotational speed over a period of 5 minutes.
  33. Send the tabulated speed and power readings to DE along with information about the axial movement of the bobbin(s).  DE will then advise on the next steps to allow rotation at the maximum operating speed.
  34. Replace the right hand side panel.
  35. Over-temperature shut down.  These shut downs are caused by a lack of cooling.  To determine the reason for the lack of cooling follow the steps below:
  36. Drive system (Inverter-Door interlock) shut down.  The drive system can stop the rotation of the instrument and the causes can be motor overload, inverter overheating or the door interlock system.  To determine the cause of the shut down the error code on the inverter will have to be read and emailed to DE.  To read the error code follow the steps below:
  37. Rotor not spinning at its maximum operational speed.  The maximum operational speed of the Mini is 2100rpm, the Spectrum 1600rpm and 1400rpm for the Midi.  The instrument not being able to reach its maximum operational speed could be an indication of worn or damaged bobbin bearing.  To determine if a bobbin bearing is worn or damaged follow the steps below:
  38. Blocked or partially blocked/crushed flying lead tubing – inspect each flying lead in for crushes especially under the flying lead clamps, replace and recheck if crushed section found.  Check the flow through each flying lead individually and replace and retest is blockage, partial blockage found.  If each flying lead is not restricting flow then the blockage in either the columns or valve box.
  39. Blocked or partially blocked columns – there has been a build up of sample residue in the column that is restricting the flow.  The solution is to follow the cleaning procedure in the instrument manual and if this does not remove the problem decontaminate the columns as described in the section Column Cleaning Procedure.
  40. Blocked or partially blocked valve box.  After following the procedure described in the above steps disconnect the valve box from HPCCC instrument.  For valve boxes fitted to the Spectrum and standard Midi follow steps i to x and for valve boxes fitted to the Mini and Midi preparative only instruments use steps xi to xviii.  Valve boxes fitted to the Mini and preparative only Midi do not have a column (Analytical/Preparative) switching valve.  When a back pressure is detected, by the pressure relief valve within the valve box opening and water flowing for the pressure relief valve waste line, the location of the blockage will be linked to the new position of the last valve to have changed position.  The blockage will either be in the valve or a connecting tube linked to the valve.  For repair of the valve box please contact DE.
  41. Broken bobbin drive belt.  If the bobbin drive belt the rotation of the bobbins is no longer synchronised with that of the rotor and all the flying leads will be damaged.  To determine if a bobbin drive belt has failed open the instrument door and see if there are pieces of a black rubber belt in the laying under the rotor.  If this is the case contact DE’s local sales agent and DE.
  42. Solvent leak from a column.  Check that the leak is not from a broken or worn flying lead and that the fittings between the flying leads and columns are tight enough to create a seal.  Once the flying leads have been checked please photograph the leak and email a copy the photograph to info@dynamicextractions.com and wait for instructions of how the leak will be repaired.
  43. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the MVB and HPCCC are as described in the operating manuals ensuring that the instrument is rotated in the “run” (clockwise) direction.
  44. Higher than normal back pressures usually indicated by release of the solvent via the pressure relief valve fitted to the system or within the valve box.  Causes:

i.Using 10cm lengths of 1.6mm bore tubing with appropriate fittings connect PC1 to PP2 on the underside of the valve box and AC1 to AP2.

ii.Replace the sample loop with another length of 1.6mm bore tubing and appropriate fittings.

iii.Switch the Injection valve to “Load”, the NP/RP valve to “RP” and the Analytical/Preparative valve to “Preparative”.

iv.Using distilled water determine if there is a back pressure at the following flow rates 25ml/min for the Spectrum and 70ml/min for the Midi.

v.Switch the injection valve to “Inject” and repeat the above step.

vi.Switch the NP/RP valve to “NP” and then repeat the step iv.

vii.Switch the Analytical/Preparative valve to “Analytical” and using distilled water determine if the there is a back pressure at a flow rate of 10ml/min.

viii.Switch the injection valve to “Load” and repeat the above step.

ix.Switch the NP/RP valve to “RP” and then repeat the step viii.

x.If this procedure does to identify the cause of the blockage within the valve box contact DE.

xi.Using a 10cm length of 1.6mm bore tubing with appropriate fittings connect Periphery to Centre on the underside of the valve box.

xii.Replace the sample loop with another length of 1.6mm bore tubing and appropriate fittings.

xiii.Switch the Injection valve to “Load” and the NP/RP valve to “RP”.

xiv.Using distilled water determine if there is a back pressure at the following flow rates 10ml/min for the Mini and 70ml/min for the Midi.

xv.Switch the injection valve to “Inject” and repeat the above step.

xvi.Switch the NP/RP valve to “NP” and then repeat the step xiv.

xvii.Switch the NP/RP valve to “RP” and then repeat the step xiv.

xviii.If this procedure does to identify the cause of the blockage within the valve box contact DE.

  1. Clear liquid inside the instrument – Condensation.  When the humidity in the environment surrounding a HPCCC instrument is high condensation can form on the heat exchange located above the rotor.  During long periods of operation the amount of condensation form a puddle of water beneath the rotor.  Such a build up of condensation will not damage the instrument.  However the puddle below the rotor should be checked to ensure that it is only water and not a mixture of solvents that has leaked for either a broken flying lead or damaged column.  If the puddle is a mixture of solvents it will smell of solvents and if it is pure water it will have no smell.  If the puddle smells of solvents check the flying leads and columns for damage and leaks, ie see the sections above.  If there is no smell remove the condensation as follows:
    1. Open the door of the instrument and using clean absorbent cloths remove the puddle of condensation below the rotor.
    2. Using more clean absorbent cloths remove the condensation that has formed on the heat exchanger above the rotor.
    3. Close the door of the instrument and it is now ready to be used.
  2. Failure of temperature control and the temperature controller displays “OPEN”.  This failure is caused by either: temperature probe damage; temperature probe being disconnected from temperature controller or corroded or broken terminations.  The resolve this follow the steps below:
  3. Open the instrument door and examine the slender silver rod that is mounted close to the rotor for obvious damage.  This silver rod is the temperature probe and if damaged explains the failure of the temperature control system.
  4. Switch off power to the instrument and then pull the front of the temperature controller out of the front of the control panel as shown in figure XX and then push in back into position to reseat the terminations.  Switch the power back on and determine if the temperature is now displayed on the front of the temperature controller.
  5. Switch off the mains power to the instrument.  Remove the right hand side panel and examine the following within the electronics box:

i.Check that all the wiring connections on the rear of the temperature controller are secure.

ii.Using an electronic multi-meter check that the electrical continuity between pin 8 on the rear of the temperature controller and pin 4 on the 36 way socket on the rear of the electronics box.  Also check that the electrical continuity between pin 9 on the rear of the temperature controller and pin 13 on the 36 way socket.

iii.Examine the sockets in positions 4 and 13 on the 36-way connector on the rear of the electronics box for damage and ensure that these sockets are at the same level as all the other sockets.  If a socket is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the electronics box.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.

  1. Measure the electrical resistance between pins 4 and 13 on the 36-way connector that was connected to the rear of the electronics box.  The resistance should be between 95 and 110 ohms, a resistance in the kW and MW ranges is an indication that there is either a high contact resistance or a break in the wiring between pins 4 and 13.
  2. Disconnect the 36-way to 24-way adaptor circuit from the case wiring loom.  Check for continuity between pin 4 on the 36-way connector and socket 7 on the 24-way connector.  Check for continuity between pin 13 on the 36-way connector and socket 8 on the 24-way connector.
  3. Examine the pins in positions 4 and 13 in the 36-way connector ensuring that these pins are at the same level as all the other pins.  If a pin is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 36-way plug.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  4. Examine the sockets in positions 7 and 8 in the 24-way connector ensuring that these sockets are at the same level as all the other sockets.  If a socket is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 24-way socket.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  5. Examine the pins in positions 7 and 8 in the 24-way plug connected to the case wiring loom ensuring that these pins are at the same level as all the other pins.  If a pin is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 24-way plug.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  6. Measure the electrical resistance between pins 7 and 8 on the 24-way plug of the case wiring loom.  The resistance should be between 95 and 110 ohms, a resistance in the kW and MW ranges is an indication that temperature probe is damaged and needs replacing.  Please contact DE for a replacement temperature probe.
  1. Electronics Box Failure.  This needs some thought!  If the front control panel is damaged during shipment from DE the electronics box which the front control panels is part of can be removed and replaced, please contact DE to arrange a replacement electronics box.  Add instructions on how to remove electronic control boxes.

Manual Valve Boxes

The range of Manual Valve Boxes (MVBs) is designed to work with the range of HPCCC laboratory instruments as described in the MVB Operating Manual, please refer to this manual.  Please ensure that the correct MVB is being used for the HPCCC instrument.  Potential faults:

  1. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the MVB and HPCCC are as described in the MVB and instrument operating manuals.
  2. Waste Lines – From the underside of a valve box there are 2 waste lines; one is connected to the sample loop and is used during sample loop filling and other is connected to the pressure relief valve.  Solvent will flow from the waste line connected to the sample loop during normal operation of a HPCCC instrument.  However solvent flowing from the pressure relief the waste line indicates a restriction in the flow path; please refer to section 10 above. 
  3. Leaks within a MVB have 3 possible causes: a loose fitting, damaged tubing/fitting or a leaking valve.  If a fitting is loose simply tighten and for other leaks please photograph the leak and email a copy of the leak to info@dynamicextractions.com and DE will organise replacement part(s).

Spectrum Automated Valve Box (AVB)

The Automated Valve Box (AVB) was developed to operate with the Spectrum HPCCC instrument and should not be connected to any other instrument as it has a pressure limit of 250psi (17bar, 1.7MPa).  Potential faults:

  1. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the AVB and HPCCC are as described in the AVB and Spectrum operating manuals.
  2. Waste Lines – The AVB has 2 waste lines; one is connected to the sample loop and is used during sample loop filling and other is connected to the pressure relief valve.  Solvent will flow from the waste line connected to the sample loop during normal operation of a HPCCC instrument.  However solvent flowing from the pressure relief the waste line indicates a restriction in the flow path; please refer to section 10 above.
  3. Leaks within a AVB have 3 possible causes: a loose fitting, damaged tubing/fitting or a leaking valve.  If a fitting is loose simply tighten and for other leaks please photograph the leak and email a copy of the leak to info@dynamicextractions.com and DE will organise replacement part(s).

Ancillaries

For a HPCCC system the major pieces of ancillary equipment are: chillers, pumps, detectors and automated valves.  The end user is responsible for using the equipment in accordance with the instruction manuals supplied and these manual are the first reference point if and when problems occur.  However, during the warranty period DE will assist in the repair/replacement of these pieces of equipment.  After the warranty period the end user is encouraged to contact the equipment manufacturer directly.

There are sundry ancillary components such as interconnecting tubing, fittings, ferrules and back pressure regulators (sprung loaded one-way valves).  During the warranty period DE will replace these items free of charge ex-works provided that the system has not been moved from where it was installed.

Chromatography Software - Clarity

DE has supplies Clarity chromatography software (produced by DataApex) for the control of all laboratory based HPCCC systems.  The end user has free life time updates and technical support from DataApex once the end user has registered their copy of Clarity with DataApex via the DataApex website (www.dataapex.com).

DE does not provide direct technical support for Clarity software but recommend that any problems are directed to DataApex.

Maxi Systems

This section to follow once the Maxi instrument and system has been developed.

Column Cleaning Procedure

This procedure uses a solution of DECON 90 and purified/distilled water to clean the columns.  DECON 90 is an emulsion of anionic and non-ionic surface active agents, stabilising agents, non-phosphate detergent builders, alkalis and sequestering agents, in an aqueous base.  If DECON 90 is not available please contact Dynamic Extractions for technical advice.

http://www.decon.co.uk/english/decon90.asp

DECON 90 is an irritant. Personal Protection Equipment (PPE) required: safety spectacles, lab-coat and gloves.

The following procedure is used to clean and decontaminate HPCCC columns.  It is intended to be used for all HPCCC instruments.

Preparation of cleaning solution

Prepare a 5% solution of DECON 90 in purified/distilled water.  Ensure that you have enough for 6 times the combined volume of the columns that are to be cleaned.

Cleaning Method

  1. Rinse column with 2 column volumes of purified/distilled water (flow rate for rinsing: analytical columns 5ml/min; for semi-preparative columns 20ml/min and preparative columns 70ml/min.
  2. Rinse column with 2-3 column volumes of 5% DECON 90 solution.
  3. Leave the cleaning solution in the columns overnight.
  4. Rinse with 2 column volumes of purified/distilled water.
  5. Rinse column with 2 column volumes of 5% DECON 90 solution.
  6. Leave the cleaning solution in the columns overnight.
  7. Rinse with 2 column volumes of purified/distilled water.

8.Repeat if contamination is still visible in rinse solution.

This procedure will leave the column clean and free of all removable contaminants.

Legal Patents

High Performance Counter Current Chromatography (HPCCC) instruments maintain high stationary phase retentions when high mobile phase flow rates are used.  These are classified as hydrodynamic instruments that operate at a g-level of 240 determined by the equation g-level = Rw2/9.81.  No other hydrodynamic instruments operate at this g-level.

Dynamic Extractions Ltd is the only manufacturer of HPCCC instruments in the world.

Dynamic Extractions Ltd protects the ability of its HPCCC instrumentation by the following granted patents:

  1. GB 2,356,365
  2. US 6,716,152
  3. Singapore patent number 109045 (WO 03//086639)
  4. GB 2,446,129
  5. US 7,351,333

Dynamic Ext

NOTE.

Please check the instruction manuals your system was supplied with as some systems may differ from standard configurations.

 

The instruments covered by this section are the Spectrum and Midi.  Potential faults:

  1. Flying leads.  Flying leads are a consumable item and need to be replaced at regular intervals.  If flying leads are not replaced regularly, fitted as described in the appropriate operating manual and maintained as described in the operating manual the flying leads will develop leaks.  To stop leaks from flying leads a programme of preventative maintenance needs to be developed in accordance with the standard practises of the user’s organisation based upon the information provided in the operating manual.  However, the following tips will help determine the potential cause of a leak:
    1. On the Spectrum check that the loops of flying lead tubing on the bobbins have not become too large and extend beyond the outer edge of the bobbin.  These loops should as shown in figures 6 and 7 in the Spectrum operating manual.
    2. Check that the fittings where the flying leads are connected to the bobbin are tight enough to create a seal.
    3. If the above steps do not uncover the cause of the leak remove the flying leads and then examine the leads to find the leak.  Once the leak has been found please photograph the damage and send the picture to DE (info@dynamicextractions.com) and then determine the following:

i.The distance between the leak and the end of the tube connected to the bobbin.  Provide DE with this measured dimension to help determine the cause of the failure.

ii.Whether the leak: is a split in the tubing or a flatten section of wear through the side of the tubing or appears to as a short cut along the length of the tubing.  Provide DE with a description of the type of leak to help determine the cause of the failure.

  1. With the information provided in the previous step DE will be able to advise upon the cause of the failure and a solution to prevent similar failures in the future and prolong the service life of flying leads.
  1. Instrument stops spinning and a red LED is flashing on the front of the instrument.  This shut down is has been caused by either an imbalance or the instrument becoming too warm.  To determine the type of failure switch the “empty-off-run” switch to the “off” position and note the temperature of the instrument.  If the temperature is greater than 45°C then an over temperature shut down has occurred.  This is confirmed by opening and closing the instrument door and the red LED continuing to flash.  If the LED continues to flash go to “Over-temperature shut down” and if the LED stops flashing go to “Imbalance shut down”.
  2. Imbalance shut down.  To remove an imbalance follow steps a to h if no valve box is fitted to the instrument and steps i to n when a valve box is fitted.
  3. Check that the flying leads are correctly labelled for the ends of the columns to which each is connected.
  4. Ensure that the columns are connected as follows: PC2 to PP1, AC2 to AP1 and then AC1 to PP2.
  5. Connect the pump to AP2 and place outlet PC1 back into the water container from which the pump is taking water.  Note the level of the water in the container.  
  6. Turn the speed control knob fully anti-clockwise.  Switch on the rotation in the “Run” (clockwise) direction and start pumping the water into the desired column at 5ml/min for a time equal to the combined column volumes divided by 5 (ml/min).
  7. This will expel any remaining air from the columns.  Now determine if the level of the water is lower in the container than at the start.
  8. Then slowly increase the rotational speed to operational speed (2100 rpm for Mini, 1600 rpm for Spectrum and 1400 rpm for Midi) over a period of 10 minutes, the imbalance should now not occur.
  9. If the instrument shuts down due to an imbalance turn the speed control knob 1 full turn anti-clockwise cancel the imbalance alarm by opening and closing the instrument door and the switch on the clockwise rotation again.  Then continue increasing the rotational speed to the operating speed again over a period of 5 minutes.
  10. Check that the flying leads are correctly labelled for the ends of the columns to which each is connected.
  11. If this does not remove the imbalance contact DE and ask for Technical Support.
  12. Ensure that the connections between the valve box and the instrument are as described in the instrument and valve box operation manuals.
  13. Switching the NP/RP valve to NP and turning the speed control knob fully anti-clockwise.  Switch on the rotation in the “Run” (clockwise) direction and start pumping the distilled water into the desired column (5ml/min for the analytical column, up to 25ml/min for the semi-preparative Spectrum column and up to 100ml/min for the Midi preparative column).
  14. When a volume of solvent equal to the column volume has been pumped into the column slowly increase the rotational speed to the operating speed over a period of 5 minutes.
  15. If the instrument shuts down due to an imbalance turn the speed control knob 1 full turn anti-clockwise cancel the imbalance alarm by opening and closing the instrument door and the switch on the clockwise rotation again.  Then continue increasing the rotational speed to the operating speed again over a period of 5 minutes.
  16. If this does not remove the imbalance contact DE and ask for Technical Support.
  17. Check that the chiller is switch on and that the chiller’s set temperature is adjusted to the usual temperature for operating the HPCCC instrument.
  18. Check that the chiller is connected to the HPCCC instrument and that there are no coolant leaks.  If there are coolant leaks contact DE for further assistance and replacement parts.
  19. Check that the coolant within the chiller is free of algae and is circulating through the HPCCC instrument.
  20. Check that temperature control on the HPCCC instrument is switched on.
  21. One the Mini and Spectrum instruments open the door of the instrument and check the cooling fan is sucking air in beneath the rotor and blowing air out above the rotor.  If the fan is not operation contact DE for further instructions.
  22. Now reduce the set temperature of HPCCC instrument to 0°C and check the speed of the cooling fan and the flow of air through the fan increases significantly.  If there is no increase in fan speed contact DE for further advice.
  23. Do not switch off the power to the instrument.
  24. Unscrew the 2-off retaining screws in the right hand side panel and then carefully lean the top edge of the side panel away for the instrument.
  25. The error code is then displayed in 3 red letters within the right hand end of the instrument.
  26. Note the error code displayed and send the code to DE via info@dynamicectractions.com  DE will use the code to determine the cause of the shut down.
  27. Reposition and secure the right hand side panel in place.
  28. Errors codes in the drive system are cleared by switching the instrument off for 30 seconds and then switching back on.  However rotation of the instrument should not be started again until you have been contacted by a representation form DE.
  29. Open the door of the instrument and determine if the bobbin(s) has axial movement of approximately 0.5 to 1mm.  For the Mini and Spectrum instruments hold the rotor still using one hand and using your other hand try to move a bobbin towards the back and front of the instrument.  For the Midi hold the rotor still and try to move a bobbin to the left and right.  If a bobbin cannot be moved or is difficult to move it can be an indication that there is a problem with a bobbin bearing.
  30. Switch off the electrical power to the instrument, remove the mains lead and alarm cable if fitted and remove the right hand side panel.  This panel is removed by unscrewing the screws that attach the panel to the case.
  31. Reconnect the mains lead and switch the electrical power back on.  The drive inverter has a red LED display on its front.  Take great care to prevent electrical shocks press the white dial on the front of the inverter and turn the dial in either direction until “SUP” appears and then press the dial.  Turn the dial again until “OPr” appears and press the dial again.  Older versions of the drive inverter use a up & down arrows and a “ENT” button.
  32. Now switch on the rotation of the instrument and set the rotational speed to maximum.  Set instrument temperature to 30°C and allow the instrument to reach this temperature and then record the reading on the inverter and rotational speed over a period of 5 minutes.
  33. Send the tabulated speed and power readings to DE along with information about the axial movement of the bobbin(s).  DE will then advise on the next steps to allow rotation at the maximum operating speed.
  34. Replace the right hand side panel.
  35. Over-temperature shut down.  These shut downs are caused by a lack of cooling.  To determine the reason for the lack of cooling follow the steps below:
  36. Drive system (Inverter-Door interlock) shut down.  The drive system can stop the rotation of the instrument and the causes can be motor overload, inverter overheating or the door interlock system.  To determine the cause of the shut down the error code on the inverter will have to be read and emailed to DE.  To read the error code follow the steps below:
  37. Rotor not spinning at its maximum operational speed.  The maximum operational speed of the Mini is 2100rpm, the Spectrum 1600rpm and 1400rpm for the Midi.  The instrument not being able to reach its maximum operational speed could be an indication of worn or damaged bobbin bearing.  To determine if a bobbin bearing is worn or damaged follow the steps below:
  38. Blocked or partially blocked/crushed flying lead tubing – inspect each flying lead in for crushes especially under the flying lead clamps, replace and recheck if crushed section found.  Check the flow through each flying lead individually and replace and retest is blockage, partial blockage found.  If each flying lead is not restricting flow then the blockage in either the columns or valve box.
  39. Blocked or partially blocked columns – there has been a build up of sample residue in the column that is restricting the flow.  The solution is to follow the cleaning procedure in the instrument manual and if this does not remove the problem decontaminate the columns as described in the section Column Cleaning Procedure.
  40. Blocked or partially blocked valve box.  After following the procedure described in the above steps disconnect the valve box from HPCCC instrument.  For valve boxes fitted to the Spectrum and standard Midi follow steps i to x and for valve boxes fitted to the Mini and Midi preparative only instruments use steps xi to xviii.  Valve boxes fitted to the Mini and preparative only Midi do not have a column (Analytical/Preparative) switching valve.  When a back pressure is detected, by the pressure relief valve within the valve box opening and water flowing for the pressure relief valve waste line, the location of the blockage will be linked to the new position of the last valve to have changed position.  The blockage will either be in the valve or a connecting tube linked to the valve.  For repair of the valve box please contact DE.
  41. Broken bobbin drive belt.  If the bobbin drive belt the rotation of the bobbins is no longer synchronised with that of the rotor and all the flying leads will be damaged.  To determine if a bobbin drive belt has failed open the instrument door and see if there are pieces of a black rubber belt in the laying under the rotor.  If this is the case contact DE’s local sales agent and DE.
  42. Solvent leak from a column.  Check that the leak is not from a broken or worn flying lead and that the fittings between the flying leads and columns are tight enough to create a seal.  Once the flying leads have been checked please photograph the leak and email a copy the photograph to info@dynamicextractions.com and wait for instructions of how the leak will be repaired.
  43. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the MVB and HPCCC are as described in the operating manuals ensuring that the instrument is rotated in the “run” (clockwise) direction.
  44. Higher than normal back pressures usually indicated by release of the solvent via the pressure relief valve fitted to the system or within the valve box.  Causes:

i.Using 10cm lengths of 1.6mm bore tubing with appropriate fittings connect PC1 to PP2 on the underside of the valve box and AC1 to AP2.

ii.Replace the sample loop with another length of 1.6mm bore tubing and appropriate fittings.

iii.Switch the Injection valve to “Load”, the NP/RP valve to “RP” and the Analytical/Preparative valve to “Preparative”.

iv.Using distilled water determine if there is a back pressure at the following flow rates 25ml/min for the Spectrum and 70ml/min for the Midi.

v.Switch the injection valve to “Inject” and repeat the above step.

vi.Switch the NP/RP valve to “NP” and then repeat the step iv.

vii.Switch the Analytical/Preparative valve to “Analytical” and using distilled water determine if the there is a back pressure at a flow rate of 10ml/min.

viii.Switch the injection valve to “Load” and repeat the above step.

ix.Switch the NP/RP valve to “RP” and then repeat the step viii.

x.If this procedure does to identify the cause of the blockage within the valve box contact DE.

xi.Using a 10cm length of 1.6mm bore tubing with appropriate fittings connect Periphery to Centre on the underside of the valve box.

xii.Replace the sample loop with another length of 1.6mm bore tubing and appropriate fittings.

xiii.Switch the Injection valve to “Load” and the NP/RP valve to “RP”.

xiv.Using distilled water determine if there is a back pressure at the following flow rates 10ml/min for the Mini and 70ml/min for the Midi.

xv.Switch the injection valve to “Inject” and repeat the above step.

xvi.Switch the NP/RP valve to “NP” and then repeat the step xiv.

xvii.Switch the NP/RP valve to “RP” and then repeat the step xiv.

xviii.If this procedure does to identify the cause of the blockage within the valve box contact DE.

  1. Clear liquid inside the instrument – Condensation.  When the humidity in the environment surrounding a HPCCC instrument is high condensation can form on the heat exchange located above the rotor.  During long periods of operation the amount of condensation form a puddle of water beneath the rotor.  Such a build up of condensation will not damage the instrument.  However the puddle below the rotor should be checked to ensure that it is only water and not a mixture of solvents that has leaked for either a broken flying lead or damaged column.  If the puddle is a mixture of solvents it will smell of solvents and if it is pure water it will have no smell.  If the puddle smells of solvents check the flying leads and columns for damage and leaks, ie see the sections above.  If there is no smell remove the condensation as follows:
    1. Open the door of the instrument and using clean absorbent cloths remove the puddle of condensation below the rotor.
    2. Using more clean absorbent cloths remove the condensation that has formed on the heat exchanger above the rotor.
    3. Close the door of the instrument and it is now ready to be used.
  2. Failure of temperature control and the temperature controller displays “OPEN”.  This failure is caused by either: temperature probe damage; temperature probe being disconnected from temperature controller or corroded or broken terminations.  The resolve this follow the steps below:
  3. Open the instrument door and examine the slender silver rod that is mounted close to the rotor for obvious damage.  This silver rod is the temperature probe and if damaged explains the failure of the temperature control system.
  4. Switch off power to the instrument and then pull the front of the temperature controller out of the front of the control panel as shown in figure XX and then push in back into position to reseat the terminations.  Switch the power back on and determine if the temperature is now displayed on the front of the temperature controller.
  5. Switch off the mains power to the instrument.  Remove the right hand side panel and examine the following within the electronics box:

i.Check that all the wiring connections on the rear of the temperature controller are secure.

ii.Using an electronic multi-meter check that the electrical continuity between pin 8 on the rear of the temperature controller and pin 4 on the 36 way socket on the rear of the electronics box.  Also check that the electrical continuity between pin 9 on the rear of the temperature controller and pin 13 on the 36 way socket.

iii.Examine the sockets in positions 4 and 13 on the 36-way connector on the rear of the electronics box for damage and ensure that these sockets are at the same level as all the other sockets.  If a socket is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the electronics box.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.

  1. Measure the electrical resistance between pins 4 and 13 on the 36-way connector that was connected to the rear of the electronics box.  The resistance should be between 95 and 110 ohms, a resistance in the kW and MW ranges is an indication that there is either a high contact resistance or a break in the wiring between pins 4 and 13.
  2. Disconnect the 36-way to 24-way adaptor circuit from the case wiring loom.  Check for continuity between pin 4 on the 36-way connector and socket 7 on the 24-way connector.  Check for continuity between pin 13 on the 36-way connector and socket 8 on the 24-way connector.
  3. Examine the pins in positions 4 and 13 in the 36-way connector ensuring that these pins are at the same level as all the other pins.  If a pin is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 36-way plug.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  4. Examine the sockets in positions 7 and 8 in the 24-way connector ensuring that these sockets are at the same level as all the other sockets.  If a socket is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 24-way socket.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  5. Examine the pins in positions 7 and 8 in the 24-way plug connected to the case wiring loom ensuring that these pins are at the same level as all the other pins.  If a pin is not at the same height as the others use long nose pliers to adjust the height by gripping the appropriate wire from inside the rear of the 24-way plug.  If a height adjustment was necessary reassemble all the connections for the temperature control system and see if the fault has been resolved.
  6. Measure the electrical resistance between pins 7 and 8 on the 24-way plug of the case wiring loom.  The resistance should be between 95 and 110 ohms, a resistance in the kW and MW ranges is an indication that temperature probe is damaged and needs replacing.  Please contact DE for a replacement temperature probe.
  1. Electronics Box Failure.  This needs some thought!  If the front control panel is damaged during shipment from DE the electronics box which the front control panels is part of can be removed and replaced, please contact DE to arrange a replacement electronics box.  Add instructions on how to remove electronic control boxes.

Manual Valve Boxes

The range of Manual Valve Boxes (MVBs) is designed to work with the range of HPCCC laboratory instruments as described in the MVB Operating Manual, please refer to this manual.  Please ensure that the correct MVB is being used for the HPCCC instrument.  Potential faults:

  1. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the MVB and HPCCC are as described in the MVB and instrument operating manuals.
  2. Waste Lines – From the underside of a valve box there are 2 waste lines; one is connected to the sample loop and is used during sample loop filling and other is connected to the pressure relief valve.  Solvent will flow from the waste line connected to the sample loop during normal operation of a HPCCC instrument.  However solvent flowing from the pressure relief the waste line indicates a restriction in the flow path; please refer to section 10 above. 
  3. Leaks within a MVB have 3 possible causes: a loose fitting, damaged tubing/fitting or a leaking valve.  If a fitting is loose simply tighten and for other leaks please photograph the leak and email a copy of the leak to info@dynamicextractions.com and DE will organise replacement part(s).

Spectrum Automated Valve Box (AVB)

The Automated Valve Box (AVB) was developed to operate with the Spectrum HPCCC instrument and should not be connected to any other instrument as it has a pressure limit of 250psi (17bar, 1.7MPa).  Potential faults:

  1. No retention of stationary phase in the attached HPCCC instrument – check that the flying leads are labelled correctly and that the connections between the AVB and HPCCC are as described in the AVB and Spectrum operating manuals.
  2. Waste Lines – The AVB has 2 waste lines; one is connected to the sample loop and is used during sample loop filling and other is connected to the pressure relief valve.  Solvent will flow from the waste line connected to the sample loop during normal operation of a HPCCC instrument.  However solvent flowing from the pressure relief the waste line indicates a restriction in the flow path; please refer to section 10 above.
  3. Leaks within a AVB have 3 possible causes: a loose fitting, damaged tubing/fitting or a leaking valve.  If a fitting is loose simply tighten and for other leaks please photograph the leak and email a copy of the leak to info@dynamicextractions.com and DE will organise replacement part(s).

Ancillaries

For a HPCCC system the major pieces of ancillary equipment are: chillers, pumps, detectors and automated valves.  The end user is responsible for using the equipment in accordance with the instruction manuals supplied and these manual are the first reference point if and when problems occur.  However, during the warranty period DE will assist in the repair/replacement of these pieces of equipment.  After the warranty period the end user is encouraged to contact the equipment manufacturer directly.

There are sundry ancillary components such as interconnecting tubing, fittings, ferrules and back pressure regulators (sprung loaded one-way valves).  During the warranty period DE will replace these items free of charge ex-works provided that the system has not been moved from where it was installed.

Chromatography Software - Clarity

DE has supplies Clarity chromatography software (produced by DataApex) for the control of all laboratory based HPCCC systems.  The end user has free life time updates and technical support from DataApex once the end user has registered their copy of Clarity with DataApex via the DataApex website (www.dataapex.com).

DE does not provide direct technical support for Clarity software but recommend that any problems are directed to DataApex.

Maxi Systems

This section to follow once the Maxi instrument and system has been developed.

Column Cleaning Procedure

This procedure uses a solution of DECON 90 and purified/distilled water to clean the columns.  DECON 90 is an emulsion of anionic and non-ionic surface active agents, stabilising agents, non-phosphate detergent builders, alkalis and sequestering agents, in an aqueous base.  If DECON 90 is not available please contact Dynamic Extractions for technical advice.

http://www.decon.co.uk/english/decon90.asp

DECON 90 is an irritant. Personal Protection Equipment (PPE) required: safety spectacles, lab-coat and gloves.

The following procedure is used to clean and decontaminate HPCCC columns.  It is intended to be used for all HPCCC instruments.

Preparation of cleaning solution

Prepare a 5% solution of DECON 90 in purified/distilled water.  Ensure that you have enough for 6 times the combined volume of the columns that are to be cleaned.

Cleaning Method

  1. Rinse column with 2 column volumes of purified/distilled water (flow rate for rinsing: analytical columns 5ml/min; for semi-preparative columns 20ml/min and preparative columns 70ml/min.
  2. Rinse column with 2-3 column volumes of 5% DECON 90 solution.
  3. Leave the cleaning solution in the columns overnight.
  4. Rinse with 2 column volumes of purified/distilled water.
  5. Rinse column with 2 column volumes of 5% DECON 90 solution.
  6. Leave the cleaning solution in the columns overnight.
  7. Rinse with 2 column volumes of purified/distilled water.

8.Repeat if contamination is still visible in rinse solution.

This procedure will leave the column clean and free of all removable contaminants.

Legal Patents

High Performance Counter Current Chromatography (HPCCC) instruments maintain high stationary phase retentions when high mobile phase flow rates are used.  These are classified as hydrodynamic instruments that operate at a g-level of 240 determined by the equation g-level = Rw2/9.81.  No other hydrodynamic instruments operate at this g-level.

Dynamic Extractions Ltd is the only manufacturer of HPCCC instruments in the world.

Dynamic Extractions Ltd protects the ability of its HPCCC instrumentation by the following granted patents:

  1. GB 2,356,365
  2. US 6,716,152
  3. Singapore patent number 109045 (WO 03//086639)
  4. GB 2,446,129
  5. US 7,351,333

Dynamic Extractions Ltd also has the following patent applications pending:

  1. PCT/GB2012/052410 also listed as WO2013/045943

ractions Ltd also has the following patent applications pending:

  1. PCT/GB2012/052410 also listed as WO2013/045943