Biotage Isolera Four User Manual Download

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411829-L, Collection and Fractionation

April 2012

Page A-5

A.2 Shoulder Slope and Valley Fractionation

A.2.1 Fractionation Using the Shoulder Slope Parameter

When the Shoulder Slope parameter is enabled (the range is 1 to 5000 mAU/CV), the Isolera
system fractionates on rising and falling shoulders and double fractionates on valleys.

Figure A-1. Fractionation on Rising Shoulder

Summary of Contents for Isolera Four

Page 1: ...Isolera User Manual...

Page 2: ...lobatch Advancer 350 AFFINILUTE Endeavor EVOLUTE ExploraSep Firefly design FLASH FlashMaster FlashVac Horizion HPFC HP SIL HP Sphere Initiator Initiator Isolera Isolera Four Isolera LS Isolera One Iso...

Page 3: ...1 17 Operational Performance Specifications 1 18 Solvent Specifications 1 19 Chemical Resistance of the Peristaltic Pump Tube Isolera LS 1 20 2 Data Administration 2 1 Log into the Data Administration...

Page 4: ...5 Configure a Network Connection 3 5 Change Report Settings 3 6 Set Up a Printer and Auto Print of Reports 3 6 Set Up File Sharing and Auto Save of Reports 3 7 Set Up Auto Send of Reports 3 8 Change...

Page 5: ...ontrol a Manual Collection 4 24 Edit and Manually Extend a Purification 4 24 Pause End or Abort a Purification 4 25 Unload a Purification 4 26 Flush a Cartridge with Air 4 26 Purge a Cartridge 4 27 Ac...

Page 6: ...nternal Detector Related Problems 6 3 Gradient Problems 6 4 Leak Detected 6 5 Overpressure Detected 6 5 7 Contact Information 7 1 Manufacturer 7 1 Biotage Sweden AB 7 1 Sales Offices and Distributors...

Page 7: ...reen G Cartridge Holder B Power Switch H SNAP Cartridge C Collection Arm I Leak Detector optional on Isolera Prime One and Four and standard on Isolera LS D Collection Rack J Leak Tray optional E Coll...

Page 8: ...d Biotage Isolera LS Large Scale The differences are listed in the table below An Isolera Prime system can be upgraded to and Isolera One or Isolera Four system An Isolera One system can be upgraded t...

Page 9: ...Chemistry Data Administration or System mode View information on the tabs and buttons in the right hand panel Scroll down or up through the text when a help window has more than one screenful of info...

Page 10: ...ternal dry loading is to be used not possible with SNAP 750g and larger cartridges the cartridge insert must be removed Internal loading can be achieved using Prepacked Samplet cartridges which are in...

Page 11: ...ay add other racks to this list see Add a Rack Type on page 2 5 Only included in the preconfigured list for Isolera Prime Isolera One and Isolera Four Not recommended on Isolera LS Only included in th...

Page 12: ...SD 1080 evaporative light scattering detector is a universal detector designed for use with Isolera One and Isolera Four systems when purifying compounds with little or no UV absorption such as carboh...

Page 13: ...Data Administration and System modes Figure 1 5 Main Menu and Data Administration Mode Isolera LS Shown 1 2 2 Data Administration Mode In the Data Administration mode the software is divided into six...

Page 14: ...se stuck check valves clean the collect valve back up and restore the database export the system configuration and logs restore the system configuration and view and reset usage statistics Network Con...

Page 15: ...inter with postscript support or a local USB printer and saved as PDF files on a USB memory device or in a share folder on your network It is also possible to create a new method with the same purific...

Page 16: ...urge Note that the Air Flush feature is not available with Isolera Prime Prime The system is priming the solvent inlet s Buttons in the Right hand Panel The following buttons are available in the righ...

Page 17: ...a share folder on the network After editing you can save the method under a new name by entering a unique method name The original method remains unaltered under its original name Base the new method...

Page 18: ...urification in progress press the Stop button It is not possible to edit a purification during the flushes and purge When using an Isolera Four system queued equilibrations take priority over queued g...

Page 19: ...nel 2 Not available if using the fixed detector Absorption measured by the external detector when connected The defined threshold in mAU The defined wavelengths and the measured absorption in mAU are...

Page 20: ...ent editor and 2 a fraction report These reports can be printed to a network printer with postscript support or a local USB printer and saved as PDF files on a USB memory device or in a share folder o...

Page 21: ...are connected to the solvents used in the method Therefore it is important that the solvents be assigned accurately If you change the solvent that is connected to a particular inlet you must make the...

Page 22: ...e collected Instruct the system to switch to a new collection vessel by clicking the New Fraction button Instruct the system to start and end an isocratic segment by clicking the Hold button isocratic...

Page 23: ...system is delivered with two kinds of peristaltic pump tubes Ensure that the used peristaltic pump tube is compatible with the solvents and sample to be used see section 1 3 4 on page 1 20 Dimensions...

Page 24: ...0 liter Rack Types 13x100 mm 16x100 mm 18x130 mm for 17 5 x 130 mm test tubes 16x150 mm 18x150 mm 25x150 mm 120 ml 240 ml 480 ml and funnel rack Not recommended on Isolera LS Cannot be used with Isole...

Page 25: ...Solvent CAS No EC No Strength1 2 Selectivity Class3 UV Cutoff nm Vapor Pressure at 20 C psi Vapor Pressure at 20 C mbar Acetone 67 64 1 200 662 2 0 56 6 VIa 330 3 6 247 4 Acetonitrile 75 05 8 200 835...

Page 26: ...d in section 5 8 1 on page 5 5 Limited The peristaltic pump tube must be discarded after each run The chemical resistance was tested in room temperature as follows The peristaltic tube was 1 weighed 2...

Page 27: ...privilege are listed in the Select User dialog 3 Select your user name If you have not been assigned a user name and or system owner privilege please contact your system supervisor 4 Press OK If your...

Page 28: ...ime To scroll a list down or up press or Figure 2 2 Cartridges Tab Isolera LS Shown 2 2 1 Add a Cartridge Type 1 Press New or if you want to base the new cartridge type on an existing one select the c...

Page 29: ...ftware s Data Administration mode all saved methods are listed You can export import and delete methods It is also possible to set default methods i e a list of methods that the users can choose from...

Page 30: ...SB memory device or in a share folder on your network if file share has been set up see page 3 7 The method s to import must be available at USB biotage isolera methods or share folder biotage isolera...

Page 31: ...base the new rack type on an existing one select the rack and press Duplicate at the Racks tab The New Rack dialog opens 2 Enter the rack settings A setting can be entered when the corresponding text...

Page 32: ...ve the changes press OK 2 4 4 Select Rack Types to List 1 If you want a rack type to appear when setting up a method in the Chemistry mode select it at the Racks tab and press Enable 2 If you do not w...

Page 33: ...nsitive 2 To export a single or all listed records as a CSV file comma delimited text file PDF file or a XML file a If you want to export the record s to a USB memory device connect the memory device...

Page 34: ...ll be used i e 200 ml min or 500 ml min 1 Neue U D HPLC Columns Theory Technology and Practice Wiley VCH 1997 2 Dean J A Lange s Handbook of Chemistry 15th edition McGraw Hill 1999 NOTE The preconfigu...

Page 35: ...ser s account when a purification has been completed An e mail will also be sent to the user when user interaction is required e g when a rack has to be replaced a solvent needs to be replenished if m...

Page 36: ...records associated with the user will be updated 4 To save the changes press OK 2 7 3 Delete a User Account WARNING All the results and methods of the user will also be deleted 1 Select the user acco...

Page 37: ...u are not at the main menu press Main Menu in the right hand panel 2 Press System All user accounts with system owner privilege are listed in the Select User dialog 3 Select your user name If you have...

Page 38: ...e unpacked and installed by an authorized Biotage service engineer Should you need to move the detector see the instructions in the ELSD 1080 Getting Started Guide P N 412941 To connect and enable an...

Page 39: ...ystem s maximum dispense rate will be used i e 200 ml min or 500 ml min 3 2 3 Disable an External Detector To disable the external detector 1 Select the Detector tab in the right hand panel 2 Select t...

Page 40: ...c When the message It is now safe to turn off the system appears on the screen turn off the system The power switch is located underneath the touch screen d Turn on the system 3 3 2 Enable or Disable...

Page 41: ...of Reports on page 3 8 Receive an e mail when user interaction is required during a purification e g when the system run out of empty collection vessels Make your reports and methods available to othe...

Page 42: ...To access the Isolera Remote Viewer 1 Enter the URL http MACHINENAME in a web browser where MACHINENAME is the hostname or the host IP address entered in the Network field above Tip You can also find...

Page 43: ...Contact your IT administrator for more information 5 Select the Paper Size text box and then select the desired paper format 6 To test the connection press Test in the Printer field If a test page is...

Page 44: ...or Disable Audible Alarm If the Audible Alarm option is enabled an audible warning will sound in the following situations 1 rack change is needed 2 leakage if using Biotage Leak Detector 3 high press...

Page 45: ...us view select the Peak Mode text box and select Yes or No 3 6 4 Enable or Disable Auto Extend If Auto Extend is enabled and the system is collecting fractions and remaining light absorption is detect...

Page 46: ...Flushes Are Performed At the end of a purification i e after the gradient purification stage is completed the system performs a Line Flush The system flushes the inlet line and the cartridge with 9 ml...

Page 47: ...x and enter the level when valley fractionation occurs 3 7 Maintenance 3 7 1 Export the System Configuration and Logs If requested by 1 Point Support to send in your system s log files follow the inst...

Page 48: ...page 3 13 3 7 3 Back Up and Restore the System s Database To back up the database 1 Select the Maintenance tab in the right hand panel 2 If you want to save the backup on a USB memory device a Connect...

Page 49: ...b in the right hand panel 2 Press Calibrate in the UV Detector field Read and follow the instructions that appear on the screen 3 7 7 Perform an Intensity Scan of the Internal Detector If requested by...

Page 50: ...el 2 Press Release Check Valves in the Valves field Read and follow the instructions that appear on the screen 3 7 10 View and Reset the Usage Statistics Both the number of runs performed on the syste...

Page 51: ...see Log into the Data Administration Mode on page 2 1 If you want to change detector network reporting and runtime settings set the system clock set the language used in the Chemistry mode enable the...

Page 52: ...up period by pressing Skip Warm Up in the right hand panel However if the UV lamp has not reached operating temperature purification results may not be optimal 4 4 Create Open and Edit Methods 4 4 1 C...

Page 53: ...or the gradient graph To show or hide the gradient table press the Table button gradient table is shown 3 In the graph the X axis indicates the length and the Y axis indicates the percentage of Solve...

Page 54: ...em With Four Solvent Inlets Shown Specify the Method Parameters at the Parameters Tab 1 Select the Parameters tab 2 Select the User text box and select your user name If you have not been assigned a u...

Page 55: ...ge the setting The software prevents you from entering a fraction volume that will overflow the vessels 5 Select the solvents to be used by selecting the corresponding text box in the Solvents field T...

Page 56: ...the Additive text box and the percentage of the additive in the solvent mix by selecting the Mix Percentage text box Not available with Isolera Prime Rack Selection Based on 13 CV gradient in Collect...

Page 57: ...25g 33 45 50g 66 85 100g 132 164 75S 264 340g 510 582 75M 528 750g 990 75L 1056 1500g 1980 13x100 mm 4 432 x 4 2 2 3 4 5 8 16 18 16 16x100 mm 4 490 x 4 2 2 3 4 5 7 14 16 15 18x130 mm 4 504 x 4 2 2 3...

Page 58: ...al Control the collection and fractionation using an external detector e g an Biotage ELSD 1080 detector Possible collection and fractionation parameters are Collect All and Start Threshold 3 If you w...

Page 59: ...n page 4 15 NOTE You do not have to save your method to be able to run it Table 4 2 Collection and Fractionation Parameters Parameter Description Start Wavelength The shortest wavelength to be include...

Page 60: ...vent B text boxes Normally the solvent with the lower solvent strength is selected as Solvent A 4 Select the Solvent Conditions text box and enter the percentage of Solvent B in the TLC solution Figur...

Page 61: ...tep Gradient editor opens 4 Enter the percentage of the strongest solvent in the TLC solution for each TLC plate by pressing the 0 table headers To add a plate press Add Plate To delete a plate select...

Page 62: ...on gradient at the Method tab press OK 13 Set up the rest of the method parameters see Specify the Method Parameters at the Parameters Tab on page 4 4 and Specify Collection Parameters at the Collecti...

Page 63: ...tected the Password dialog opens Enter your password using the keypad and press OK 5 To run a purification using the method press Run 6 Prepare the system as described on page 4 15 7 Proceed with step...

Page 64: ...displayed 6 When you are pleased with the result press Save to Editor The method is opened at the Method tab and can be edited and or saved 4 4 5 Create a Method Using a Web Browser See Access the Iso...

Page 65: ...of the correct solvent is present in each solvent reservoir and that the waste reservoir has sufficient capacity for the run To assign solvents and set reservoir capacities and current levels see page...

Page 66: ...411829 L Operation April 2012 Page 4 16 4 Figure 4 10 Tube Connections...

Page 67: ...with the sample loading pump To empty the pump tubing of liquid and mount a new peristaltic pump tube 1 Place the pump s outlet line into a waste reservoir 2 Select the Solvents tab in the right hand...

Page 68: ...choice 2 Place the pump s outlet line into a waste reservoir Figure 4 13 Sample Loading Pump 3 Select the Solvents tab in the right hand panel in the software and then the Sample Loading Pump tab 4 T...

Page 69: ...xt run not recommended clean the tube with methanol as soon as possible Note that the sample loading pump can be cleaned while a purification is in progress For more information and instructions see p...

Page 70: ...ns take priority over queued gradient runs Figure 4 15 Run Parameters and Estimation Dialogs Figure 4 16 Status Tab Isolera LS and Isolera Four EXP with Spektra License Shown Isolera One EXP and Isole...

Page 71: ...00 SNAP HP 340g 510 340 6800 13600 13600 17800 17800 34000 65 300 SNAP Ultra 10g 17 10 200 200 1000 1000 2000 10 50 SNAP Ultra 25g 45 25 500 500 2500 2500 5000 20 100 SNAP Ultra 50g 85 50 1000 1000 50...

Page 72: ...and remaining light absorption is detected when the system nears the end of a purification the system enters the Auto Extend mode if enabled in the system mode This extends the gradient purification s...

Page 73: ...wn on the horizontal axis Full When enabled all readings on the horizontal axis are shown Length Unit Toggle the horizontal axis unit between CV minutes and milliliter UV Zero Used to set the current...

Page 74: ...old button again 4 7 6 Control a Manual Collection If you have set up a purification with manual collection you can instruct the system to start and stop collecting fractions by pressing the Collect A...

Page 75: ...the software The collection arm returns to its home position the inner right corner and the system is paused 2 To resume the run from the point at which it was paused press Resume in the right hand p...

Page 76: ...the rack s used by the run Allocated racks are listed at the Allocated heading at the Status tab 5 If using an Isolera Four system with automatic rack allocation enabled load new empty racks of the sa...

Page 77: ...lush with the weakest solvent used in the run To change select the Solvent text box and select the desired solvent 7 Ensure that a sufficient quantity of the selected solvent is present in the solvent...

Page 78: ...tings Note that the result filter is case sensitive If you want to list all result records press Clear All 6 To search press Search If there are records matching your search criteria they are listed i...

Page 79: ...sted see Search for Records above An archive report with purification details a chromatogram a 3D absorbance spectrum for the whole detector range requires a Spektra license and TLC data if entered in...

Page 80: ...e same color Select All Select all fractions Deselect All Deselect all fractions Modify how the 3D absorbance spectrum is presented in the archive report See step 3 above Length Unit Change the horizo...

Page 81: ...stored as individual records in the system s database The records can if desired be saved by the user as XML files text files including all raw data on a USB memory device or in a share folder on the...

Page 82: ...itoring is enabled select the Capacity text box and enter the capacity of the solvent reservoir 6 If solvent monitoring is enabled select the Current text box and enter the current solvent level of th...

Page 83: ...e solvents are assigned accurately to the solvent inlets at the Instrument Solvents tab If the capacity and current volume values are available for the waste and or solvent reservoirs i e the monitori...

Page 84: ...pense stroke the maximum flow rate when using an Isolera Prime system is equal to Max Fill Rate x Max Dispense Rate Max Fill Rate Max Dispense Rate Examples If the fill rates for the used solvents are...

Page 85: ...atic hold is enabled Instruct the system to pause the purification in progress by clicking the Pause button For safety reasons it is only possible to pause and not resume a run from your office Instru...

Page 86: ...please see the Biotage Product Catalog The catalog can be downloaded at www biotage com For a complete list of accessories please contact you local representative 5 2 1 All Isolera Systems Accessory...

Page 87: ...VLV 1000 Sample injection valve 3 way to SNAP or SNAP Ultra cartridge adapter 1 411081 Biotage ELSD 1080 external detector 1 ISO ELSD 1080 Accessory Quantity Part Number Sample pump tube PharMed limit...

Page 88: ...surfaces using a clean lint free cloth lightly dampened with water If required a small amount of mild soap may also be used After cleaning wipe dry with a clean lint free cloth 5 5 Implement a Mobile...

Page 89: ...ow cell P N 09869 for Isolera Prime One and Four systems and P N 412324 for Isolera LS systems and perform an absorbance calibration see Calibrate the Internal Detector on page 3 13 6 Reconnect the tu...

Page 90: ...istent dispensing volumes can be signs of a dirty collect valve Use the following procedure to clean the collect valve 1 Select the Maintenance tab in the right hand panel 2 Press Clean Collect Valve...

Page 91: ...age 4 17 5 9 Sonicate Solvent Inlet Filters A solvent inlet filter is installed on the end of each solvent inlet line The solvent inlet filters protect the pump and cartridges from damage due to parti...

Page 92: ...tem as follows 1 If the Leak Detected dialog is open close it by pressing Close 2 If a purification is in progress press Stop at the Status tab and then Abort in the Stop dialog 3 If a prime is in pro...

Page 93: ...he power outlet 3 Remove the spillage using the appropriate safety precautions 4 If using Biotage Leak Detector ensure that the leak detector and the leak tray including the space underneath the detec...

Page 94: ...nect the power cord from the power outlet and contact Biotage 1 Point Support see contact details on page 7 1 5 10 4 Assembling Tubes Correctly WARNING Shut down the system before replacing any tubing...

Page 95: ...sen the fuse holder by carefully prying under the notch on the left side of the holder with a small standard flat blade screwdriver see Figure 5 8 Figure 5 8 Loosen the Fuse Holder 5 Grasp the fuse ho...

Page 96: ...ce panel at the left side of the system using a Torx 10 screwdriver 4 Loosen the screw locking the UV lamp in a vertical position see Figure 5 12 using a Torx 20 screwdriver Figure 5 12 Loosen the Loc...

Page 97: ...e tungsten lamp and disconnect it from the power cable see Figure 5 15 c Mount a new tungsten lamp Ensure the lamp is fully inserted and the power cable is positioned so that it does not get in contac...

Page 98: ...vailable at www biotage com 6 3 Fraction Collector Related Problems Problem Possible Causes Solutions 1 The collection arm does not position correctly over each collection vessel a Misaligned rack s E...

Page 99: ...t is not recommended Particulate free solvent is required If the problem persists the pump piston seals or the check valve seals may be worn Please contact Biotage 1 Point Support see contact details...

Page 100: ...t is high absorbing at the selected wavelength s Change the collection and fractionation wavelength s or turn the Baseline Correction option on if using a system with a Spektra license installed c Lam...

Page 101: ...the cartridge and reach the detector the initial front of the gradient will always be delayed by at least 1 CV compared to the programmed gradient A longer gradient delay may be due to interactions w...

Page 102: ...Prime at the Prime tab and start a new prime with a lower flow rate e If the overpressure occurred during an equilibration or a gradient run a Press Edit at the Status tab The Edit Method dialog opens...

Page 103: ...otage com When contacting 1 Point Support please have the following information ready Company name and contact information Serial number of your Isolera system see the product label at the rear of the...

Page 104: ...Fractionation Methods The following fractionation methods are supported Manual Manual fractionation Volume Fractionation when a test tube or bottle is full based on the specified max fraction volume f...

Page 105: ...collection and fractionation reaches the specified Start Threshold level Fractionations occur when the other signal reaches the Start Threshold level and then when one of the signals decreases below...

Page 106: ...V1 UV2 or all The measurement results in a baseline containing the maximum absorbance of the solvent system at each wavelength used a solvent absorbance spectrum During the gradient run the maximum so...

Page 107: ...available with the UV1 and UV1 UV2 detection modes Table A 3 System Parameters Parameter Description Start Slope Enable The level where the parameter Start Slope is enabled Below this level the Start...

Page 108: ...y Fractionation A 2 1 Fractionation Using the Shoulder Slope Parameter When the Shoulder Slope parameter is enabled the range is 1 to 5000 mAU CV the Isolera system fractionates on rising and falling...

Page 109: ...411829 L Collection and Fractionation April 2012 Page A 6 A Figure A 2 Fractionation on Falling Shoulder Figure A 3 Double Fractionation on Valley...

Page 110: ...2 Fractionation Using the Valley Slope Parameter When the Valley Slope parameter is enabled the range is 1 to 5000 mAU CV while the Shoulder Slope parameter is disabled Off the Isolera system only fr...

Page 111: ...411829 L Collection and Fractionation April 2012 Page A 8 A A 3 Collection and Fractionation Examples A 3 1 Threshold Collection A 3 2 Slope Collection...

Page 112: ...411829 L Collection and Fractionation April 2012 Page A 9 A A 3 3 Slope Collection and Threshold Fractionation A 3 4 Valley Fractionation...

Page 113: ...variable or UV VIS detector is used with the UV1 UV2 detection mode both detector signals used for collection and fractionation are checked for a possible auto extend of the gradient Collection Metho...

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