introduction to the hplc chemstation and acquisition

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  • Slide 1
  • Introduction to the HPLC ChemStation and Acquisition
  • Slide 2
  • 2 In This Section, We Will Discuss: How to work in the Microsoft Windows Environment The structure of the ChemStation Software. How to set up an acquisition method. How to run a single sample.
  • Slide 3
  • 3 l Delete temporary files on a regular basis. Use Clean Disk for Windows 2000 and XP. l Use Checkdisk to find and correct errors on the disk. l Defragment the hard drive. l Use Virus detection software. l Create an Emergency Repair disk. Maintaining the Computer System Accessories Right-click drive letter
  • Slide 4
  • 4 The HPLC ChemStation Software Add Instruments Schedule ChemStation Tasks Access Instrument and Software
  • Slide 5
  • 5 Method and Run Control View Sampling Diagram System Diagram Online Plot Navigation Pane ChemStation Explorer Navigation Buttons
  • Slide 6
  • 6 ChemStation Explorer Views
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  • 7 ChemStation Views Views Method and Run Control Data Analysis Report Layout Verification (OQ/PV) Diagnosis Full menu or Short menu Change Views
  • Slide 8
  • 8 View Preferences Allows you to configure the contents of the ChemStation Explorer Specifies naming convention for sequence data containers Specifies how signals are loaded
  • Slide 9
  • 9 Parts of a Method l Method information l Instrument control parameters l Data analysis parameters l Run Time Checklist What is a Method? A method comprises all the parameters necessary to perform data acquisition and data analysis, including integration and calibration parameters, for one sample. Pre- and post-run tasks may be specified by a command or macro in the run-time checklist. The method is identified by a file name with a.m extension. Master methods are stored in Chem32\#\Methods.
  • Slide 10
  • 10 Instrument Parameters and Control: System Diagram or Menus Click on GUI for parameters. Instrument control via menus or GUI Click here for instrument control.
  • Slide 11
  • 11 Create a Method In the Method and Run Control view, Select New Method, or double-click on DEF_LC.M. DEF_LC.M is loaded. This method is a template file that cannot be overwritten.
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  • 12 You may use Edit Entire Method to sequentially move through instrument parameters required to acquire data for one analysis or access the parameter windows by selection. Note: Edit Entire Method does not access all instrument parameters such as More Pump > Auxiliary, etc. Editing a Method Using Edit Entire Method
  • Slide 13
  • 13 Select Portions of Method to Edit Information about the method Instrument parameters found in Method and Run Control view Parameters for post-acquisition processing found in Data Analysis Parts of the method to run
  • Slide 14
  • 14 Method Information Fill in any information you want stored with the method.
  • Slide 15
  • 15 Time programmable composition, flow, and pressure. Pump Parameters
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  • 16 Injector Parameters - Agilent 1100/1200 Standard Sample capacity l 100 x 2 ml vials in 1 tray l 40 x 2 ml vials in 1/2 tray l 15 x 6 ml vials in 1/2 tray l Microvials with sleeves Injection volume l 0.1 - 100 l standard l Up to 1500 l with multi-draw kit. l Up to 900 l in a single draw using expanded injection upgrade kit.
  • Slide 17
  • 17 l NEEDLE WASH to reduce carry-over to the absolute minimum l MULTI DRAW MODE for injection volumes greater than 100 ul l SWITCH VALVE TO BYPASS to decrease standard loop delay volume (300ul) to a minimum (bypass) delay volume of 6.2 ul l INJECTOR PROGRAM for programming custom injection steps Widest dynamic injection range: 0.1 l-1.5 ml From pump To column Metering device To waste 4-port rotor seal Agilent 1100/1200 Injector Special Functions
  • Slide 18
  • 18 Agilent 1200 High Throughput Samplers Sample Capacity 2 well plates (96 and 384) plus 10 additional 2-mL vials. 108 x 2-mL vials in 2 x 54 vial plate plus 10 additional 2-mL vials. 30 x 6-mL vials in 2 x 15 vial plate plus 10 additional 2-mL vials. 54 Eppendorf tubes (0.5/1.5/2.0mL) in 2 x 27 Eppendorf tube plate. Also compatible with the Agilent 1200 Series sample capacity extension for further expansion of the sample capacity.
  • Slide 19
  • 19 Agilent 1100/1200 DAD Parameters 5 signals (standard DAD). 8 signals (SL). Sample signal 191 - 949 nm. Slit programmable; 1, 2, 4, 8 and 16 nm settings. Time programmable. 80 Hz data rate DAD (SL) for rapid resolution columns. DAD SL Window shown
  • Slide 20
  • 20 VWD Parameters Time Programmable Set peak width to narrowest chromatographic Peak width. VWD G1314C
  • Slide 21
  • 21 Column Thermostat Parameters l 10 degrees below ambient to 80 degrees C (Standard). l 10 degrees below ambient to 100 degrees C (SL). l Two separate heated zones for two columns. l Optional valve for column switching applications. l Compartment holds up to 30 cm column. l Column identification module with injection record for GLP. (TCC SL shown)
  • Slide 22
  • 22 Run Time Checklist Select items to execute during the method. Send your report to Excel using a custom macro.
  • Slide 23
  • 23 Saving the Method Save a method by selecting Save Method or Save Method As from the Method menu, or select the Save Method Tool.
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  • 24 Prepare the Instrument UV Lamp On Turn on a UV lamp at least 20 minutes prior to your first analysis for warm-up by clicking the control button. Balancing Ready
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  • 25 Prepare the Instrument Prime the Pump Purge Valve 1.Make certain the vacuum degasser is on (if applicable). 2.Open the purge valve. 3.Pump 5 mL/min of 100 % A until all air bubbles have cleared.
  • Slide 26
  • 26 Prepare the Instrument Prime the Pump 4.Pump 5 mL/min at 100%B until all air bubbles have cleared. 5.Pump 5 mL/min at 100 % for each remaining channel. 6.Change the composition to that of your next run and continue. 7.Change the flow rate to that of your next run. 8.Close the purge valve.
  • Slide 27
  • 27 Prepare the Instrument Instrument Actuals Allows you to review your instrument parameters, check pump pressure and module status at a glance.
  • Slide 28
  • 28 Prepare the Instrument - Instrument Actuals
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  • 29 Edit Signal Plot Click Change
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  • 30 Run One Single Injection To inject an individual sample, select Sample Info..., then Run Method.
  • Slide 31
  • 31 Start Method
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  • 32 Follow Acquisition
  • Slide 33
  • 33 Logbook Entries Check how the run proceeded in the Logbook.
  • Slide 34
  • 34 Directory Structure for Data Files Instrument # UV Spectra UV Signal Chromatograms Logbook for Run
  • Slide 35
  • 35 Turn Off System Remember to flush buffers from the system. Do not leave 100% Acetonitrile in the system. Do not leave the TCC at high temperatures without column flow for extended periods.


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