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HPLC-ECD Stand Alone System

(HTEC-500)

User’s

Guide

Eicom Corporation

AC01-0319

Summary of Contents for HTEC-500

Page 1: ...HPLC ECD Stand Alone System HTEC 500 User s Guide Eicom Corporation AC01 0319...

Page 2: ...njector and the Autosampler Electrical Signal Connection 6 6 Online Elution Degassing System 7 6 1 If you think the degassing efficiency is reduced you may diagnose the problem by the following proced...

Page 3: ...on 22 11 1 Basics 22 11 2 Working Electrode 23 11 3 Applied Potential 23 11 4 Time Constant 25 11 5 Background Current 25 11 6 If Background Current is High 25 11 7 Caution for the Platinum Working El...

Page 4: ...This page intentionally left blank...

Page 5: ...ve the outlet screw and aspirate all the remaining mobile phase in the detector cell with a syringe Pay attention to prevent any tubing from becoming blocked Salt solvents must be rinsed out by ultrap...

Page 6: ...gasser Column Clips ECD Cell Door Cabinet Thermo Controller Control Panel ECD Terminal Ch1 Tubing Guide Terminal Ch2 for PEC 510 Tubing Guide Cell Mount Fig 2 HTEC 500 Front View Door open Status Indi...

Page 7: ...is for the nut Flat Seal Nut Two way Joint Tubing connection Connect the tubing and keep its tip sticking out of the ferrule as seen above except for the connection with the degasser which uses a flat...

Page 8: ...This may raise the background current of the ECD When all reagents are added plug the bottle and shake and mix it thoroughly Check if all reagents are dissolved Filtration is good for removing unwant...

Page 9: ...jectors of the autoinjector online autoinjector EAS 20 and or the autosampler should be set up downstream to the manual injector For the autoinjector connect no 3 port of the manual injector with no 2...

Page 10: ...eadings see chapter 8 page 12 for the errors This way you can prevent loosing samples when the HPLC pump HTEC 500 accidentally stops Please follow this diagram to figure out the electrical connections...

Page 11: ...e liquid in the degassing component At a flow rate of 500 l min of mobile phase the degassing efficiency is the same between the two channels There is no independent switch to turn on and off the dega...

Page 12: ...ase container on the same level as the HTEC 500 Pump Outlet Piston Wash Port Check Valve Outlet Pump Head Piston Wash Port Check Valve Inlet nut PEEK Tubing 0 75 mm ID Easy Fit Tubing connected to the...

Page 13: ...g 9 Display PUMP Press 0 64MPa FlowRate 200 L min P u l s e Fr e e M o d e O N P r e s s L i m i t 1 4 M P a Indicates the pump is running Pressure status Setting flow rate Use the 10 keys for setting...

Page 14: ...latency of this convergence is shorter at higher flow rates B Flow Rate Compensation Always turn the pulse free mode off before starting maintenance exchange of mobile phase purging and so on because...

Page 15: ...Disconnect the pump inlet tubing at the outlet of the degasser 3 Run the pump 4 Open the pump drain valve and aspirate mobile phase slowly by using a syringe from the drain port so that the pump inle...

Page 16: ...m the tubing after the pump When aspirating from the purge drain using a syringe the disconnected tubing which was connected to the column must be plugged or air will be introduced through the aspirat...

Page 17: ...ot cleared check the pressure differences of each part as you did for the precolumn above by going from downstream to upstream of the part This way you should identify the part which is blocked Cut th...

Page 18: ...hase bottle is on the same level as the HTEC 500 Please remove the filter if you have one D One of the check valves has lint dust or something inside This problem happens more frequently in the inlet...

Page 19: ...s that have unusual pressure changes and cause the pump motor to generate too much torque If this problem happens after a few rotations of the pump motors and it does not stop immediately after starti...

Page 20: ...ble generates inside of the syringe please flush out the solution with the air bubble and slowly re aspirate Generally the knob is set at INJECT before and after the injection B Insert the syringe nee...

Page 21: ...ampler autoinjector Leave the manual injector at the inject position while you are using the other injectors so that the sample loop on the manual injector can be kept clean by the mobile phase If the...

Page 22: ...GEL 4 6 mm ID The PC 04 is also used with the PP ODS 4 6 mm ID and the precolumn is positioned before the injector but not just before the separation column This is because there are no compatible pac...

Page 23: ...ure of the pump If the precolumn generates more than 2 MPa please compare with and without precolumn the upstream filter must be exchanged and the precolumn must be repacked Repacking of a precolumn r...

Page 24: ...he precolumn with the packing material before the inside of the precolumn becomes dry Avoid any gap in the precolumn Fill up the syringe with fresh methanol after disposing of the methanol collected d...

Page 25: ...gh 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 The filter can be disco...

Page 26: ...cally active substances are eluted from the separation column and then electrolyzed by the working electrode in the electrode system A current is produced by electrolysis and this is in proportion to...

Page 27: ...nce Applied potential is set in the following way Push the display key until the ECD screen appears Push the edit key until is displayed in front of applied potential Input the target potential with t...

Page 28: ...and then draw the shaft out The cell is now open 3 Lift and remove the setting plate working electrode and gasket in turn 4 Carefully wipe the surface of the electrode smoother side with acetone usin...

Page 29: ...there is less background noise and more stability 11 6 If Background Current is High The surface of the working electrode could be dirty In this case disassemble the cell see Fig 23 and wipe the surfa...

Page 30: ...methanol Then set the position of the injector to INJECT and let the 30 methanol flow for 1 hour through the line at 70 or 80 of the set flow rate value This value is specific to the type of separatio...

Page 31: ...ndard in cold storage with 0 1 M CH3COOH containing 1 mg mL EDTA 2Na at a concentration of 1 ng L Dilute using 20 mM phosphate buffer pH3 5 containing 20 mM EDTA 2Na for every analysis It decomposes e...

Page 32: ...500 microdialysis sample injections If you find odd shaped peaks it may be time to repack the precolumn To judge the quality of the precolumn remove it and connect a separation column directly to the...

Page 33: ...tem 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 Injector Seals If the injector is used continuously...

Page 34: ...everal hours or stays high Possible causes are as follows The separation column and or precolumn are new Please wash the columns with clean analytical mobile phase overnight The wrong mobile phase con...

Page 35: ...f a standard sample is lower than usual Possible causes are as follows The standard has decomposed oxidized The standard is not stable when its concentration is low and it should be prepared just befo...

Page 36: ...ollows The precolumn is contaminated Inspect the cause of the high pump pressure by disconnecting the tubing from downstream to upstream When the pressure of the tubing is high the inlet of the tubing...

Page 37: ...emove both nuts 4 To remove the pump head slowly and carefully pull the pump head directly towards you without twisting it Vertical rotation as piston becomes axis Other movement in other directions w...

Page 38: ...screws being careful not to break or scratch the piston Do not lose the back up ring This is a khaki colored part which supports the piston seal from the back side of the pump head Please do not forg...

Page 39: ...o the seal Water in the pump loosens the seal and pushes it up Please note that the pump internal volume is sometimes too small to push the seal easily If the seal does not come up please try to pour...

Page 40: ...be a different color Fasten the screw securely to set the piston in place Then wipe the surface of the piston with acetone using a Kimwipe to remove oil from your fingers and dust 18 Carefully place b...

Page 41: ...y 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 Any movement in any direction other than directly straight in...

Page 42: ...p r o C m o c i E 113 Kita enmenden cho u k i m i h s u F a b o t o m i h S Kyoto Japan 612 8497 Amuza Inc 10060 Carroll Canyon Rd Ste 100 San Diego CA 92131 Phone 888 680 7775 Fax 858 560 8040 Websit...

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