NOTE: Reaction Analytics Inc. supports the integration of iChemExplorer to a variety of HPLCs from different suppliers.  The iChemExplorer software is compatible with proprietary and open data formats.  Please contact Reaction Analytics Inc. to determine how the iChemExplorer may be made compatible for you application.  The information below refers to the integration of iChemExplorer with the Agilent HPLC.

The methods to control operation of the HPLC are created and stored in the HPLC Control application such as Agilent ChemStation.  The following guidelines refer to the creation and editing of methods in Agilent ChemStation for use by iChemExplorer.  

To create suitable methods - Open Agilent ChemStation.  Select method.  Edit method.  Click through the Edit Method windows to make the following changes...

• Select one wavelength to collect data.  Selection of multiple wavelengths will be visualized as overlapping peak profiles.
• In the Destination section select File.  Change Prefix to ichem.  In the File Type section, select .WMF (.EMF for 32 bit version) and .CSV.  These are the file types used by iChemExplorer.

Save the method as edited.  These changes may be saved under a new method name.  All method files should be saved to the Methods folder in ChemStation as this is the default target from which iChemExplorer populates the method list in the iSample tab.

To create a sampling sequence with iChemExplorer - Power up the modules of the HPLC.  Open Agilent ChemStation On-Line version.  Confirm that all modules are on line - icons in green.  Open iChemExplorer.  Select the iSample tab.

• Start by selecting from the Sampling Method drop down list.  Changing methods will reset all other entries.  Selecting the method after making other entries will reset these entries.
• Select the vials to sample from the RV list or with the tool - red box - on the tray grid.  Remember that each vial should be sampled at least once every two hours.  The HPLC will cycle through each vial before returning to the first vial.  If the run time per vial is five minutes then no more than 24 vials should be in the run.  This is a good reason to consider changes to minimize run time - different columns, higher operating pressure, higher column temperature, etc.
• Enter the sample volume in microliters in the box next to Sample Injection Volume.  Choose the default if unsure.  Remember that the sample is taken at full concentration of the reaction.  The sample volume should be as low as possible so as not to overwhelm the detector.
• Enter Time in minutes for the sample sequence to run.  The time should be greater than the expected activity of the chemistry.
• Select Edit Sequence Param to check ChemStation set-up for data collection.  The Sequence Parameters from ChemStation is opened.  In the Data File section, click auto (for auto sequencing of the folder name with each injection by vial and sample sequence).  Create a new subdirectory for the chemistry.  In the Shutdown section, click Post Sequence Cmd/macro.  From the drop-down select macro SHUTDOWN.MAC.go.  This shuts off the HPLC at the end of the run time.
• Select Transfer to ChemStation.  A sequence table is created in ChemStation from the entries in iChemExplorer.  Maximize ChemStation to check the sequence table.  Make adjustments to the sequence table such as adding transfer methods for dosing from one vial to another, adding wait steps for slow chemistry such as solubility testing and so on.
• SEE the Start Sequence button.  This initates the run of the sequence table.  You may wish to wait for the start until the tray heating and stirring have been started.  See below.

To control heating and stirring - Select the iHeat-iStir tab.  Check the Comm entry.  If "n.c." for not connected then select box for drop down and select appropriate port to communicate to Heater Stirrer Module - usually COMM1.  Check that the external probe provided with the Heater Stirrer Module is in a vial in the tray and that the vial is filled with a heat transfer fluid - silicone oil is recommended.  Heating and stirring may be controlled by the ramp and hold sequence table or on-the-fly. 

For on-the-fly (manual) control select the box next to on-the-fly.  A window opens.  Use the slides to set temperature and stirring.  Select the sensor for temperature feedback control in the Control by section - default is to external probe with choice of sensor in the hotplate.  Use the drop down to Shutdown control after the selected time elapses.  Select Start Control button to initiate heating and stirring and check Log Data to create a record of temperature and stir rate over time.  Entries for temperature will be made periodically and at the time a sample is taken from a vial.

For for the ramp/hold table, use the three boxes across the top to enter elapsed time in minutes, temperature and stir rate at the end of the elapsed time.  The iChemExplorer software makes a linear extrapolation from the conditions at the prior set points to the new set points by the end of the elapsed time.  For a ramp, provide a short elapsed time - 3 to 5 minutes - with temperature and stir rate.  To hold, enter a long elapsed time - 60, 120 or 180 minutes - and keep temperature and stir rate the same.  Select the Add Temp/Stir to add these entries to the table.  The software checks that the elapsed time for each succeeding entry is greater than the previous entry.  To delete entries, higlight the entries and select the Delete Temp/Stir.  Adds may only be added to the end of the ramp hold table.  To start heat and stir control for the table, select the Start Ramp button.  This button DOES NOT start the sequence table for the HPLC. 

REMEMBER to add a last entry to set temperature and stir rate to 0 as the hotplate will remain at the last values in the table at the end of the run.

To view data in real time - Select the iGraph tab.  Go to Real Time profiling and select On.  Select Load HPLC Data button to open Windows Explorer window.  Select subdirectory for run data.  Open folder.  Select *.csv folder and select Open.  The iGraph tab window will be automatically populated with the peak profile.  Move cursor to window and left click to open window with chromatograph.

NOTE: The record of temperature and stir rate over time is logged to templog.csv and stored to the subdirectory for the chemistry data folders.