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Eicom HTEC-500 User Manual

Hplc-ecd stand alone system
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AC01-0319
HPLC-ECD Stand Alone System
(HTEC-500)
User's Guide
Eicom Corporation

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Summary of Contents for Eicom HTEC-500

  • Page 1 AC01-0319 HPLC-ECD Stand Alone System (HTEC-500) User’s Guide Eicom Corporation...

  • Page 2: Table Of Contents

    Contents Important Checklist Before Operation ..........1 Terminology ..................2 A List of Fitting Parts and Directions for Use ........3 Mobile Phase Preparation ..............4 System Installation ................5 5-1. Wiring ..................5 5-2. Tubing and Degasser ............... 5 5-3.
  • Page 3 Manual Sample Injector and Autoinjector ........16 9-1. Structure ..................16 9-2. Operation of Manual Injector ..........16 9-3. Autosampler and Autoinjector Setup ........17 Precolumn .................... 18 10-1. Types of Precolumn Housing and Line Filters ......18 10-2. Identification of Precolumn Quality........19 10-3.
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  • Page 5: Important Checklist Before Operation

    This can be the cause of noise in the ECD. • Place the mobile phase container and the waste container on the same level as the HTEC-500. • Use glass bottles as mobile phase and ultrapure water containers. Do not use any plastic containers.

  • Page 6: Terminology

    Tubing Guide HPLC Pump (See Fig. 7) Tubing Guide Online Elution Degasser ECD Cell Cell Mount Thermo Controller Fig. 2 HTEC-500 Front View (Door open) Main Power Switch (Back Side) Status Indicator Cabinet Drain Fig. 3 HTEC-500 Front View (Door closed)

  • Page 7: A List Of Fitting Parts And Directions For Use

    HTEC-500 A List of Fitting Parts and Directions for Use Tough Connector Nut for Inlet of Cell Low pressure part. For column connections. Hex Nut High pressure part (pumps) Ferrule for Inlet of Cell The narrow side is for the cell and the wide side is for the nut.

  • Page 8: Mobile Phase Preparation

    Prepare the mobile phase according to the Eicom technical information. The mobile phase preparation depends on what you are measuring with the HTEC-500. Please read below to find a general method for mobile phase preparation. Also, the following points should be noted because the ECD has a higher sensitivity compared to other detectors: •...

  • Page 9: System Installation

    Tubing and Degasser Pull the tubing’s for the pump rinse, the mobile phase suction and the cell drain out of the HTEC-500 system. Put all the equipment on a level place. Mobile phase and waste bottles should be placed on the same level as the HTEC-500 (See Fig.

  • Page 10: Connecting To The Autoinjector And The Autosampler

    A trigger signal needs to be sent from the online autoinjector or the autosampler to the data acquisition unit to inform it of the injection timing. It also needs to be sent to the HTEC-500 in order to give the auto zero signal.

  • Page 11: Online Elution Degassing System

    HTEC-500 Online Elution Degassing System The degassing component works to remove dissolved air from the mobile phase. The dissolved air occasionally interferes with the HPLC pumping or electrochemical detection. To prevent unexpected problems with the analyses, always use the degassing system. Mobile phase goes through the system when the pump is running and it is automatically degassed.

  • Page 12: Hplc Pump Operation

    • The mobile phase container should be a clean glass bottle. Do not use a plastic container. • Put the mobile phase container on the same level as the HTEC-500. PEEK Tubing 0.5 mm ID...

  • Page 13: Run/Stop

    HTEC-500 7-1. Run/Stop Use the pump key to turn the pump on and off. When the pump is running, the pump light should be on at the front of the system. Also, *PUMP* on the liquid crystal display (LCD) indicates the pump is running, while -PUMP- indicates the pump is off.

  • Page 14: Pulse Free Mode Setting

    Function of Pulse Free Mode A. Pulsation Quenching The HTEC-500 is equipped with a pulse free mode. This is a pulse quenching system. A computer detects pulsatile flow as the pressure changes (this information is provided by a pressure sensor) and controls the pistons movements during each stroke but does not influence the stroke cycle or the flow rate.

  • Page 15: How To Fix Pump Errors And Problems

    HTEC-500 How to Fix Pump Errors and Problems 8-1. Less Flow Rate, Less Pump Pressure or Unstable Pump Pressure There are a few possibilities to resolve these problems. Please try A to C as follows: A. Remove Air from Pump Heads and Check Valves If the pressure is unstable or lower than normal for the set flow rate, a small trapped air bubble in one of the pump heads or the check valves could be the cause of this problem.

  • Page 16: Types Of Pump Errors

    HTEC-500 B. Remove Air from Tubing Going to the Column If these above steps don’t work, another cause of this problem could be that an air bubble(s) may be trapped in the tubing between the pump and the (pre)column. To solve this, disconnect the tubing upstream of the precolumn and pump mobile phase through until the air bubble(s) comes out.

  • Page 17: How To Fix These Errors

    HTEC-500 8-3. How to Fix these Errors A push of the pump key clears the errors. If the same error occurs repeatedly, the problem can be resolved by referring to the following: Error 1 The pressure limit value may be set too low. If so, raise the limit value. The optimum pressure limit is different for each column (see Table 2).
  • Page 18 7). The cause of resistance in the pumping system can be either that the mobile phase bottle is at a lower position compared to the HTEC-500 or an inline filter is present upstream of the pump (typically a tubing anchor filter in the mobile phase bottle is used). Check that the mobile phase bottle is on the same level as the HTEC-500.
  • Page 19 If this problem happens without the motor rotation after running the pump and if the pump stops immediately after pressing the pump key, it is because of the failure of the power train or the pressure sensor of the pump. Please contact Eicom to fix the power train.

  • Page 20: Manual Sample Injector And Autoinjector

    Whenever injecting into the manual injector, use the accessory syringe. If you need to inject more than 25 μl, please order the 50 μl syringe (Eicom model number 705SNR). Do not connect any other tubings to the drain port. This is to avoid any potential problems.

  • Page 21: Autosampler And Autoinjector Setup

    9-3. Autosampler and Autoinjector Setup When you hook up an autoinjector or autosampler to the HTEC-500, do not remove the manual injector from the system. If you ever have problems with the HTEC-500+Autosampler/Autoinjector, the manual injector will be needed to diagnose which part is the problem.

  • Page 22: Precolumn

    In this case, the packing material should be chosen according to the kind of separation column being used. If not, the shape of the peaks may be influenced. For more information, please refer to Eicom reports.

  • Page 23: Identification Of Precolumn Quality

    2 MPa (please compare with and without precolumn), the upstream filter must be exchanged and the precolumn must be repacked. Repacking of a precolumn requires the precolumn packing tool set. Select the precolumn packing material according to Eicom technical notes. See also Table 3. If Using the PP-ODS Column It is difficult to know if the precolumn should be repacked because the sample is not introduced into the precolumn in the case of microdialysis.

  • Page 24: How To Repack

    HTEC-500 10-3. How to Repack Upstream Disconnect the precolumn as shown below and Flow take the packing material away. Then wash the inside of the precolumn with running water and then rinse it with ultrapure water. For every two times you repack, please exchange the upstream filter once as described in chapter 10-4, page 21.

  • Page 25: Filter Exchange

    HTEC-500 10-4. Filter Exchange If you try to suck methanol through the precolumn with a syringe and it does not do so smoothly, this indicates that the upstream filter (PF-04 for 4 mm ID, PF-03 for 3 mm ID) is blocked and it should be exchanged.

  • Page 26: Ecd Operation

    HTEC-500 ECD Operation Attention • The tip of the reference electrode should be soaked in mobile phase or lithium chloride solution. • The reference electrode needs to be replaced to a new one once a year, even if it is not used.

  • Page 27: Working Electrode

    HTEC-500 11-2. Working Electrode There are six types of working electrodes. Please select the working electrode based on the Eicom technical information. Each application/technical note provides a guide for the working electrode selection. The gasket (GS-25) has a 25 micron meter thickness and this allows for higher sensitivity.
  • Page 28 HTEC-500 In this photo, the ECD cell is disconnected from the ECD but it’s not necessary for normal use 1. Loosen the outlet screw and insert a syringe. It’s easier if the outlet tubing is disconnected. Suck the liquid from inside the ECD cell. Without this...

  • Page 29: Time Constant

    Generally, when the lower theoretical platelets column is selected, select a longer time constant. When using Eicom applications, please follow the application notes. 1.0 sec is rarely used. Please select 1.5 sec or 3.0 sec for our applications.

  • Page 30: Storage Of The System

    HTEC-500 Storage of the System The method of storage depends on how long the storage is for. 12-1. Short-term (A few days to a few weeks) Just switch off the system and leave it running in mobile phase. Wash behind the pumps with ultrapure water.

  • Page 31: Storage Of A Standard Sample

    HTEC-500 Storage of a Standard Sample Substances that are active to the ECD are easily oxidized and should be stored carefully. Check the conditions where they are stable. The methods to store a standard sample for some typical substances are described below: Catecholamine: Put the standard in cold storage with 0.1 M HCl containing 1 mg/mL EDTA-2Na at a...

  • Page 32: Regular Maintenance Schedule

    Eicom user’s manuals. If you find that the installed gasket is dusty or the edge of it is not clearly cut, please replace it immediately. If you are using a graphite electrode and it has been exposed to more than 35% methanol or 15% acetonitrile, please change it.
  • Page 33 Pistons and seals for maintenance are provided separately from the HTEC-500 and require special tools which are provided with the system. Please see Appendix A for a protocol for how to replace pump parts. Eicom also provides a maintenance service. Please contact us for more details.

  • Page 34: Troubleshooting

    Please use the manual injector of the HTEC-500 to inject samples and only inject standard samples to diagnose the problem at first. Do not use the autosampler to diagnose the problems listed below. Once you find that the HTEC-500 is OK, please start to use the autosampler again.
  • Page 35 • The drain port of the injector is blocked. • Some unwanted bacteria may have generated in the HTEC-500. Change the tubing and fill up the degasser with hypochlorous acid overnight and then wash away using an excess volume of ultrapure water.
  • Page 36 • The piston seal or the piston is worn. It must be changed for a new one. You may observe lower pressure than before. • The setting temperature of the cabinet of the HTEC-500 is lower than before. • Contamination on the precolumn. Please repack.

  • Page 37: Appendix. A

    HTEC-500 Appendix. A Piston and Piston Seal Exchange Note: Please read each step completely before you proceed. Also, please remember the order and orientation of all parts as you remove them. This helps to avoid having to disassemble the pump and risk breaking or damaging the piston.
  • Page 38 HTEC-500 6. Remove the pump wash port connectors. If 8. Pull the pump head mount (made of stainless your machine was purchased before June steel and held by fingers in the picture below) 2006, you do not need to use the wrench as straight out after removing all the screws, shown in the picture.
  • Page 39 HTEC-500 10. Remove the piston and look at it under bright 13. The seal should come up like in this picture. If light. If the reflection of the piston is different this step proves unsuccessful, please pull the at the area around the piston seal, it needs to seal carefully using pliers.
  • Page 40 HTEC-500 16. Now remove the tool. 18. Carefully place back the pump mount. Please pay attention to the orientation of the parts. The wider screw holes should be on the top and the bottom as seen below. 19. Do not forget to put on the back-up ring.
  • Page 41 HTEC-500 21. Reassemble the pump head carefully. Take care not to break the new piston by twisting the pump head. After insertion of the piston slowly and softly into its position at the back of the pump head, please push the center of the pump head a little bit harder, but carefully, using a finger.
  • Page 42 Amuza Inc 10060 Carroll Canyon Rd. Ste 100 113 Kita enmenden-cho San Diego, CA 92131 k - i Phone: 888-680-7775 Kyoto, Japan 612-8497 Fax: 858-560-8040 Website: www.eicomusa.com...

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