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CombiFlash® R

f

Installation Guide

2-2

WARNING

The system is heavy. Use a two-person lift to prevent injury.

Before making any connections to the Combi

Flash

R

f

, place

the system on the bench or in the fume hood where it will be
operated. Temporarily position the system so you can access
the back panel to complete the connections.

2.3 Connect Power

Ensure that the On/Standby switch below the touch screen
panel is in the Standby position. Then, use the supplied IEC
power cord to connect the Combi

Flash

R

f

to mains power.

WARNING

Mains power must meet the voltage, frequency, and amperage
requirements listed on the serial number label.

2.4 Connect Solvent Lines

WARNING

Risk of fire ignited by electrostatic discharges. Never
substitute the black tubing on CombiFlash systems. The black
PTFE tubing (P/N 023-0503-06) is conductive. This tubing is
required to dissipate static electricity.

CAUTION

To prevent damage or premature wear to the pump and internal
valves, clean solvent should be used. The solvent should not
contain any dissolved solids.

The Combi

Flash

R

f

has four solvent inlet ports on the back

panel: two A solvents and two B solvents. The system requires
at least one A and one B solvent to form a binary gradient, and
assumes that the B solvent is stronger than A. The second set
of inputs allow you to form alternative binary gradients, or
change the A or B solvent mid-run.

Accessory package (P/N 60-5239-006) includes several pieces
to complete the solvent inlet connections.

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Summary of Contents for Installation Guide

Page 1: ...llation Guide For indoor use only COPYRIGHT 2002 2004 by Teledyne Isco Inc 4700 Superior St Lincoln Nebraska U S A 68504 Phone 402 464 0231 Toll Free 800 228 4373 FAX 402 465 3022 Revision A August 15 2007 Part 60 5233 385 ...

Page 2: ...e Unit 2 1 2 2 Instrument Location 2 1 2 3 Connect Power 2 2 2 4 Connect Solvent Lines 2 2 2 5 Connect Waste Lines 2 5 2 6 Optional External Gas 2 9 2 7 Connect and Route Drain Lines 2 9 2 8 Position the System 2 9 2 9 Install Collection Tube Racks 2 10 2 10 Turn on Power 2 12 2 11 Configure the System 2 12 2 11 1 Solvents 2 12 2 11 2 General Settings 2 13 2 11 3 Vapor Limit 2 13 2 11 4 Automatica...

Page 3: ...Access 4 2 4 1 2 Network Printing 4 3 4 1 3 Automatically Print Report at End of Run 4 4 4 2 Direct Connection 4 4 4 2 1 Windows 2000 Settings 4 5 4 2 2 Windows XP Settings 4 6 4 2 3 Windows Vista Settings 4 8 4 2 4 Completing the Direct Connection 4 10 5 Maintenance 5 1 Preventive Maintenance 5 1 5 2 Cleaning 5 1 5 2 1 Collection Rack and Tray Cleaning 5 2 5 3 Tubing Inspection 5 2 5 4 Flow Cell ...

Page 4: ...llation guide contains this guide in Adobe Acrobat PDF format and the PeakTrak On line Help in PDF format 1 2 Product Overview The Teledyne Isco CombiFlash Rf flash chromatography system gives you high productivity automation programmable gradients full spectrum UV detection and peak separation and automatic detection of columns and collection tube racks Its small size makes it a great personal sy...

Page 5: ...Rf system at the instrument from a computer in the lab or from remote locations The CombiFlash Rf is optimized for use with Teledyne Isco s RediSep Rf columns which are pre packed with a variety of media For example the CombiFlash Rf can purify samples from 4 milligrams to greater than 33 grams using RediSep Rf columns loaded with 4 to 330 grams of silica gel Applications include purification of o...

Page 6: ...trol from a computer on an established network Note Teledyne Isco recommends that you obtain assistance from your Information Technology department before attempting direct or network connections to a Windows computer 1 3 1 Multiple Control Possibilities The system can be accessed from the built in touch panel and up to ten network computers The touch panel shares control with all connected comput...

Page 7: ...connected to power 95 relative humidity maximum at 20 to 40 C Flow Rate Range 5 to 100 ml min Flow Rate Accuracy 5 Pressure Limit Up to 200 psi column dependant For columns undetected by RFID the maximum pressure is limited to 50 psi Pressure Accuracy 5 of full scale Peak Detection Modes Slope or threshold UV Spectrum 200 to 360 nm Wavelength Accuracy 5 nm Flow Cell Pathlength 0 1 mm 25 Fraction A...

Page 8: ...ort that accepts USB Flash memory drive A Flash memory drive may be inserted into this port for transferring files importing and exporting methods and system software updates 3 Adjustable Column Mount The injection valve assembly and upper column mount slide along this mount so the system can accept a variety of column sizes 4 On Standby Switch 1 On 0 standby 5 Fraction Collector Arm and Drop Form...

Page 9: ...les 12 Column Expansion Ports These ports allow the system to add column channels with an optional module This option is not available at the time of printing 13 Upper Drain Tube Liquids spilled on the top shelf are carried away through this tube to a user supplied container 14 Ethernet Port An 8P8C jack for a network connection using a standard CAT5 cable or for a direct connection to a computer ...

Page 10: ...Section 1 Introduction 1 7 Figure 1 3 CombiFlash Rf Features Side Figure 1 4 CombiFlash Rf Features Back 11 12 19 20 23 16 13 14 15 22 21 18 24 17 ...

Page 11: ...te port Inject Valve Waste The injection valve is cleaned after each run The system directs fluids from this automatic cleaning routine to this port 24 Lower Drain Tube This tube carries away liquids spilled onto the rack area 1 6 Safety Before installing operating or maintaining this equipment it is imperative that all hazards and preventive measures are fully understood While specific hazards ma...

Page 12: ...gulations In the event of a solvent or chemical spill your organization should have a plan to deal with these mishaps In all cases use good laboratory practices and standard safety procedures 1 6 1 Hazard Severity Levels This manual applies Hazard Severity Levels to the safety alerts These three levels are described in the sample alerts below CAUTION Cautions identify a potential hazard which if n...

Page 13: ...ll be seriously injured if you place them between the moving parts of the mechanism near these symbols Symboles de sécurité Ce symbole signale l existence d instructions importantes relatives au produit dans ce manuel Ce symbole signale la présence d un danger d électrocution Risque de pincement Ces symboles vous avertit que les mains ou les doigts seront blessés sérieusement si vous les mettez en...

Page 14: ...telbar drohenden Verletzungsgefahr für Finger und Hände wenn diese zwischen die beweglichen Teile des gekennzeichneten Gerätes geraten Advertencias y Precauciones Esta señal le advierte sobre la importancia de las instrucciones del manual que acompañan a este producto Esta señal alerta sobre la presencia de alto voltaje en el interior del producto Punto del machacamiento Sus dedos o manos seriusly...

Page 15: ...orized representative immediately WARNING If there is any evidence that the system has been damaged in shipping do not plug it into AC power Contact Teledyne Isco or its authorized representative for advice Compare the contents of the boxes with the enclosed packing slip If there are any shortages contact Teledyne Isco immediately 2 2 Instrument Location The CombiFlash Rf has a relatively small fo...

Page 16: ...nnect Solvent Lines WARNING Risk of fire ignited by electrostatic discharges Never substitute the black tubing on CombiFlash systems The black PTFE tubing P N 023 0503 06 is conductive This tubing is required to dissipate static electricity CAUTION To prevent damage or premature wear to the pump and internal valves clean solvent should be used The solvent should not contain any dissolved solids Th...

Page 17: ... and larger opening on the cap Figure 2 1 the other has no plate and a smaller opening on the cap Figure 2 2 Figure 2 1 Recommended solvent connection with plate 60 0923 017 Ferrule 60 0923 015 Compression Fitting Nut 60 5234 130 Label 1 of 4 60 5235 042 38 mm Cap or 291 0010 06 45 mm Cap 60 5233 399 38 mm Plate or 60 5233 400 45 mm Plate 60 5234 129 Inlet Filter Assembly Place weighted filter at ...

Page 18: ... Ensure that the solvent and air tubing are secured so that the weighted filter remains at the bottom of the solvent source 60 0923 017 Ferrule 60 0923 015 Compression Fitting Nut 60 5234 130 Label 1 of 4 60 5235 050 38 mm Cap or 60 5235 051 45 mm Cap 60 5234 129 Inlet Filter Assembly Place weighted filter at bottom of container To Solvent Level Sensing To Solvent Supply Connection ...

Page 19: ... decreased flow due to the high vapor pressure of DCM This problem becomes more pronounced as the ambient temperature increases 2 5 Connect Waste Lines WARNING Risk of fire or equipment damage Failure to connect Waste Port tubing may allow organic solvents to pool in unsafe areas possibly creating dangerous levels of flammable vapors WARNING Risk of fire ignited by electrostatic discharges Never s...

Page 20: ...plete the connections according to Figure 2 3 or 2 4 If your connection hardware matches Figure 2 3 connect the Waste Level Sense line to the compression fitting in the center of the cap Push the ends of the two black waste lines through the holes in the cap If your connection hardware matches Figure 2 4 push the three tubes through the cap Push the stop on the Waste Level Sense tubing against the...

Page 21: ...ions with metal plate 60 0923 017 Ferrule 60 0923 015 Compression Fitting Nut 60 5234 143 Waste Tubing Assembly 60 5234 051 38 mm Waste Cap or 60 5234 142 45 mm Waste Cap To Diverter Valve Waste Connection To Waste Level Sense Connection To Inject Valve Waste Connection ...

Page 22: ...015 Compression Fitting Nut 60 5234 143 Waste Tubing Assembly 60 5235 053 38 mm Waste Cap or 60 5235 054 45 mm Waste Cap To Diverter Valve Waste Connection To Waste Level Sense Connection To Inject Valve Waste Connection Tubing Stop Ventilation Hole Luer fitting If necessary connect user supplied tubing and route to fume hood ...

Page 23: ...in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components The CombiFlash Rf has drain tubes extending from its top shelf and back panel The tubes drain away any liquid spilled on the top shelf and the tray beneath the fraction collection racks Route the end of this tubing to a suitable waste fluid collection cont...

Page 24: ...s for 25 150 mm vials 50 mL Total vials 60 60 5237 040 One rack for 500 mL French square bottles To load the racks 1 Insert test tubes vials or bottles into the rack Fig 2 5 CAUTION Risk of broken glass or equipment damage Do not load test tubes longer than the length listed on the tube size label 2 While holding the rack with the tube size label visible Figure 2 6 insert the racks into the system...

Page 25: ...n the system will automatically detect the type of rack and configure program settings accordingly Later in section 2 11 6 you can configure the maximum volume for each tube size Figure 2 5 Loading test tubes Figure 2 6 Inserting the collection tube rack Tube Size Label ...

Page 26: ...uration tab for the desired operation and then click OK to save the settings Network Configuration settings are discussed later in Section 4 Remote Interfaces 2 11 1 Solvents When you installed the system up to four solvents were connected to the back panel This section of the Configuration window allows you to manage these solvents To do so Note The system is shipped with common solvent names alr...

Page 27: ... Name optional Use this option to name your system The name will appear in operational displays and run summaries This feature is useful when your laboratory has more than one CombiFlash Rf system Language Select the desired language from the drop down list box 2 11 3 Vapor Limit The system has an internal vapor sensor that detects vapors present in the immediate atmosphere When the limit is excee...

Page 28: ...n and a connection to a printer on the network Network settings are discussed in Section 4 Remote Interfaces Do not select this option for these initial installation steps 2 11 5 Enable Run Length Extension When enabled this option automatically extends the run if a peak is eluting at the end of the maximum B gradient This ensures that a late eluting peak fully comes off the column and is collecte...

Page 29: ...e minutes at the minimum B 2 11 6 Set Default Tube Volumes Click this button to open a window from which you can set the default volume of the collection tubes You can increase or decrease these values to set the desired default volume collected in each container Just be sure you don t overflow the containers by exceeding their capacity The system will automatically advance to the next tube when t...

Page 30: ...duration in minutes will vary according to the column media and size and the flow rate 2 11 9 Gradient Method Select the standard gradient type Figure 2 8 Linear The system gradually mixes solvents A and B from the last programmed B until it reaches the next programmed B This method connects the inflection points that define the gradient with a straight line Step The system holds each B value unti...

Page 31: ... first time inspect the solvent and waste connections to the system If any leaks are observed click the Cancel button to stop the Auto Prime Correct the leak by tightening the fitting an additional 1 4 turn and then restart the Auto Prime from step 1 To advance to the next Auto Prime step you can click the Fast Forward button The Auto Prime window closes when finished After Auto Priming the system...

Page 32: ...n 3 1 2 Solid Samples Some compounds are not soluble in solvents that are compatible with the chromatography In addition the compounds may have very limited solubility in any solvent resulting in sample volumes that are impractical for good chromatography The solution in this case is solid sample introduction Here the reaction mixture or dissolved sample is mixed with the same media in the chromat...

Page 33: ... the sample to adsorb to the silica d Remove the solvent with a suitable method such as rotary evaporation 3 Load the media and sample mixture into the cartridge Tap the cartridge on the benchtop to settle the mixture 4 Place a frit on the top of the cartridge Force the frit down against the mixture using the plastic plunger 5 Wipe any residual powder inside the neck of the cartridge Empty solid s...

Page 34: ...ction 3 3 for more details 3 2 Loading a RediSep Rf Column To load a RediSep Rf column Note For best results always use RediSep Rf columns The system will not automatically detect other columns including non Rf RediSep columns The system also limits the maximum operating pressure to 50 psi when the column is not detected 1 Select a RediSep Rf column The on line help includes a column selection gui...

Page 35: ... Default methods are optimized for the use of RediSep Rf columns The default settings will equilibrate the column collect all fluid in the fraction collector rack differentiate between UV vs non UV absorbing fluid and place the fluids into separate tubes cut UV absorbing peaks based on slope detection or level threshold To start the run from the PeakTrak Main window 1 Enter a sample name If you ch...

Page 36: ... the column The system will skip column equilibration so that the sample will not be flushed from the column before the run 5 Review or update the Start Rack and Start Tube 6 Review the Minimum Run Requirements The system will report the estimated solvent volumes expected waste collection tube usage etc You can use this information to verify that there will be enough solvent to complete the run th...

Page 37: ... the system equilibrates the column and then waits When instructed perform the following steps a Place the injection syringe on the injection port b Slowly force the liquid into the injection port c Chase the first injection with a second injection solution of at least 0 25 mL The chase solution can be the starting mobile phase or diluted sample mixture d Click OK to continue the run 3 4 During th...

Page 38: ... Next Tube This button advances the fraction collector to the next tube position This allows you to conveniently collect elute of interest in a new tube Note When you click the Next Tube button the system immediately marks the tube advance on the chromatogram However the system delays the movement of the drop former over the tubes This delay accounts for the volume held in the tubing between the d...

Page 39: ...ed gradient for the entire run length When this step is complete the system will purge the column with air and clean the injection valve Clicking the Fast Forward button will cause the system to skip any remaining time in the current step and advance to the beginning of the next step Unlike the Rewind button the clicking the Fast Forward button to complete the run will not modify the current metho...

Page 40: ...ceeding The system has a factory assigned IP address 192 168 1 51 In many cases this address must be reconfigured for use on a corporate network To reconfigure the IP address 1 From the touch screen panel select the Tools Configuration menu command 2 The Configuration window has two tabs Select the Network Configuration tab 3 Select the Network Type The options are Static IP and DHCP Static IP If ...

Page 41: ...e DHCP request and will be used to update the DNS settings This name would allow you to assign an Instrument Name such as My_Rf Users would then access the instrument through the address http My_Rf 4 Click the OK button to save the settings 5 Confirm that your network administrator has completed any necessary network changes to support the system 6 Locate the CAT5 connection cable P N 480 6545 01 ...

Page 42: ...st also find out the queue name If the printer uses a JetDirect print queue also ask for the port number When this information is known you can proceed with configuring system for network printing 1 Select the Tools Configuration menu command 2 The Configuration window has two tabs Select the Network Configuration tab 3 Follow the on screen instructions for entering the address and queue informati...

Page 43: ... window has two tabs Select the Instrument Configuration tab 3 Select the Automatically Print Report at End of Run option 4 Click OK to save the setting 4 2 Direct Connection A direct connection supports communication between the CombiFlash Rf system and a single PC that meets the recommendations in Table 4 1 This guide provides instructions for Windows 2000 section 4 2 1 Windows XP 4 2 2 and Wind...

Page 44: ... Local Area Connection Properties Window will open Figure 4 1 4 Highlight the Internet Protocol TCP IP item in the list and click on the Properties button The Internet Protocol Properties Window will open Figure 4 2 Note The following steps change the local area connection settings If the PC must be restored to its original network configuration record the present Internet Protocol TCP IP settings...

Page 45: ...gure 4 1 Local Area Connection Properties Window Figure 4 2 Internet Protocol Properties Window 4 2 2 Windows XP Settings 1 From the Windows Start button open the Control Panel 2 Locate the Network Connection icon and open this control panel ...

Page 46: ...lect Properties Refer to Figure 4 3 4 Highlight the Internet Protocol TCP IP item in the list and click on the Properties button Refer to Figure 4 4 5 Select the Alternate Configuration tab Click the User Configured option See Figure 4 5 6 Enter the IP address 192 168 1 200 Figure 4 3 LAN or High speed Internet Connection Figure 4 4 Local Area Connection Properties ...

Page 47: ...Settings 1 From the Windows Start menu right click on Network then select the Properties menu option The PC displays the Network and Sharing Center 2 Under Tasks select Manage Network Connections 3 Right click on the Local Area Connection icon and select the Properties option After you provide permission to continue the PC displays the Local Area Connection Properties window Figure 4 6 4 Select In...

Page 48: ...Section 4 Remote Interfaces 4 9 Figure 4 6 Local Area Connection Properties Figure 4 7 Internet Protocol Version 4 TCP IPv4 Properties ...

Page 49: ...s window Click the OK button to close the Local Area Connection properties window 9 Shut down the PC and place the CombiFlash Rf system in Standby 4 2 4 Completing the Direct Connection 1 Connect the PC to the CombiFlash Rf system using the cross over cable 2 Turn the system s On Standby switch to ON Wait for the PeakTrak operating screen to appear on the CombiFlash Rf system then start the PC 3 S...

Page 50: ...ntage of solvent B to flush residual compounds from the column internal tubing and flow cell Refer to section 5 4 1 Nightly Log off the system File Log out and place the On Standby switch in Standby Monthly Perform these tasks at least monthly more frequently if conditions warrant Tubing Inspection section 5 3 Collection rack and tray cleaning section 5 2 1 Monthly flow cell cleaning section 5 4 3...

Page 51: ...y Use distilled water with a mild detergent For tougher stains use isopropyl alcohol 5 3 Tubing Inspection WARNING Risk of fire or equipment damage Faulty tubing fittings and drains may allow organic solvents to pool in unsafe areas creating a potential for dangerous levels of flammable vapors Improper draining may damage the instrument s internal components Perform a tubing inspection monthly 1 V...

Page 52: ...hole Figure 5 2 Top shelf drain hole Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the collection tray drain hole Vacuum or pressurized air applied to the outlet end of the drain tube must exist at the top shelf drain hole ...

Page 53: ...he last compound elutes Skipping the flush may cause residual compounds to build up and crystallize which might result in cross contaminating later separation runs higher operating pressures reduced flow cell lamp energy a noisy baseline on the absorbance trace frequent messages recommending flow cell cleaning Figure 5 4 Allow the separation run to complete the full default column method If the co...

Page 54: ...message you can Cancel Run recommended Click the Cancel Run button so you can perform a quick cleaning described in the following steps Continue Collect All Click this button to ignore the message Because the peak detection operation might be impaired the system automatically collects all fluids to avoid diverting compounds of interest to waste Help Click this button to display the flow cell clean...

Page 55: ...ght be a normal indication for systems with flow cell windows stained by compounds Green lamp energy is sufficient to detect peaks up to 3 0 Absorbance Units 3 Remove the column and insert a bypass tube between the upper and lower column mounts A bypass tube P N 209 0165 46 can be found in the accessory kit P N 60 5239 006 4 Set the Flow Rate to 40 mL min Figure 5 6 5 Select Through column for the...

Page 56: ...dicator reaches the far right of the green range When the indicator is in the yellow the lamp energy and flow cell are ready for the next run However continuing the cleaning for greater raw lamp energy should reduce the frequency of flow cell cleaning messages Figure 5 4 If after five minutes the indicator is still in the red range repeat the cleaning steps using an alternative solvent Table 5 1 O...

Page 57: ...izing organic molecules Dimethylsulfoxide DMSO Strong solvent that will clear organic sample Dimethylformamide DMF Strong solvent if other solvents do not produce the desired results Tetrahydrofuran THF Strong solvent Water DMF methanol A 50 25 25 solvent mixture effective in clearing some obstructions Water DMF Acetic Acid 30 30 30ml solvent mixture effective in clearing some obstructions Mildly ...

Page 58: ...ion option 5 Select Next for the Pump into Tube option 6 Place the B1 Solvent inlet line into a reservoir of methanol acetone or a strong solvent that readily dissolves residual sample material 7 Click the Prime B button to pump 100 Solvent B through the bypass tube and into the collection tubes 8 After three minutes click the Stop button Allow the system to stand for at least six hours Overnight ...

Page 59: ...cord connection If the line cord is connected properly check the fuses in the power input module located on the system s rear panel These are time delay fuses T type and rated according to the mains input power voltage 100 to 129 VAC mains input 2 0 A fuse 215 to 264 VAC mains input 1 0 A fuse CAUTION Always replace fuses with the same type and rating ...

Page 60: ...e Criteria EN61000 4 2 Electrostatic Discharge Level 2 4kV contact discharge Level 3 8kV air discharge A EN61000 4 3 Radiated RF Immunity 80 MHz to 1000MHz 80 AM at 1kHz Level 2 3V m A EN61000 4 4 Electrical Fast Transient Level 2 1kV on AC lines A EN61000 4 5 Surge on AC Lines Level 2 1kV common mode Level 2 0 5KV differential mode A EN61000 4 6 Conducted RF on AC lines 150 kHz to 80 MHz Level 1 ...

Page 61: ...duct O ST O Represent the concentration of the hazardous substance in this component s any homogeneous pieces is lower than the ST standard limitation X ST X X Represent the concentration of the hazardous substance in this component s at least one homogeneous piece is higher than the ST standard limitation Manufacturer may give technical reasons to the X marks The Environmentally Friendly Use Peri...

Page 62: ...r the Teledyne Isco trademark or for which any other warranty is specifically stated Damage to CombiFlash instruments resulting from the failure of a non RediSep column may cause the need for repairs that incur billable service activities The customer is responsible for these charges This warranty is expressly in lieu of all other warranties and obligations and Teledyne Isco specifically disclaims...

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