Introduction%20to%20the%20HPLC%20ChemStation%20and%20Acquisition

1
Introduction to the HPLC ChemStation and
Acquisition
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.

3
Maintaining the Computer System

• Delete temporary files on a regular basis. Use
  Clean Disk for Windows 2000 and XP.
• Use Checkdisk to find and correct errors on the
  disk.
• Defragment the hard drive.
• Use Virus detection software.
• Create an Emergency Repair disk.

Accessories
Right-click drive letter
4
The HPLC ChemStation Software
Add Instruments
Access Instrument and Software
Schedule ChemStation Tasks
5
Method and Run Control View
System Diagram
ChemStation Explorer
Sampling Diagram
Online Plot
Navigation Pane
Navigation Buttons
6
ChemStation Explorer Views
7
ChemStation Views

• Views
• Method and Run Control
• Data Analysis
• Report Layout
• Verification (OQ/PV)
• Diagnosis

Change Views
Full menu or Short menu
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

9
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.

Parts of a Method

• Method information
• Instrument control parameters
• Data analysis parameters
• Run Time Checklist

10
Instrument Parameters and Control System
Diagram or Menus
Click on GUI for parameters.
Click here for instrument control.
Instrument control via menus or GUI
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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.

12
Editing a Method Using Edit Entire Method
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 gt Auxiliary, etc.
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
14
Method Information
Fill in any information you want stored with the
method.
15
Pump Parameters
Time programmable composition, flow, and pressure.
16
Injector Parameters - Agilent 1100/1200 Standard

• Sample capacity
• 100 x 2 ml vials in 1 tray
• 40 x 2 ml vials in 1/2 tray
• 15 x 6 ml vials in 1/2 tray
• Microvials with sleeves
• Injection volume
• 0.1 - 100 ?l standard
• Up to 1500 ?l with multi-draw kit.
• Up to 900 ?l in a single draw using expanded
  injection upgrade kit.

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Agilent 1100/1200 Injector Special Functions

• NEEDLE WASHto reduce carry-over to the absolute
  minimum
• MULTI DRAW MODEfor injection volumesgreater
  than 100 ul
• SWITCH VALVE TO BYPASS to decrease standard loop
  delay volume (300ul) to a minimum (bypass) delay
  volume of 6.2 ul
• INJECTOR PROGRAMfor programming custom injection
  steps

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.

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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
20
VWD Parameters
VWD G1314C
Set peak width to narrowest chromatographic
Peak width.
Time Programmable
21
Column Thermostat Parameters

• 10 degrees below ambient to 80 degrees C
  (Standard).
• 10 degrees below ambient to 100 degrees C (SL).
• Two separate heated zones for two columns.
  
• Optional valve for column switching
  applications.
• Compartment holds up to 30 cm column.
• Column identification module with injection
  record for GLP.

(TCC SL shown)
22
Run Time Checklist
Select items to execute during the method.
Send your report to Excel using a custom macro.
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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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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Prepare the Instrument Prime the Pump

1. Make certain the vacuum degasser is on (if
   applicable).
2. Open the purge valve.

Purge Valve

1. Pump 5 mL/min of 100 Auntil all air bubbles
   have cleared.

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Prepare the Instrument Prime the Pump

1. Pump 5 mL/min at 100B until allair bubbles have
   cleared.
2. Pump 5 mL/min at 100 for eachremaining channel.

1. Change the composition to thatof your next run
   and continue.
2. Change the flow rate to that of yournext run.
3. Close the purge valve.

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Prepare the Instrument Instrument Actuals
Allows you to review your instrument parameters,
check pump pressure and module status at a glance.
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Prepare the Instrument - Instrument Actuals
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Edit Signal Plot
Click Change
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Run One Single Injection
To inject an individual sample, select Sample
Info..., then Run Method.
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Start Method
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Follow Acquisition
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Logbook Entries
Check how the run proceeded in the Logbook.
34
Directory Structure for Data Files
Instrument
Logbook for Run
UV Spectra
UV Signal Chromatograms
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.