DIRECTIONS FOR USING THE HP 1090 SERIES L

LIQUID CHROMATOGRAPH

Sylvia Sid and Margaret V. Merritt

Modified by Susan J. Kohler

October 19, 1998

 

A. Starting up the system: NOTE IF SYSTEM HAS BEEN STARTED FOR YOU, START WITH SECTION D.
  1. Turn on in order: HP 1090 Series L liquid chromatograph (power switch is located in back of the machine about midway up the left side), printer, monitor, disk drive, and computer. Wait for prompt on the computer screen.
  2. On the computer keyboard, enter the correct Date [Return] and Time [Return].
  3. (You should now be see Top Level displayed at the lower left corner. This corner will always display what program or mode the computer is in. The bottom of the screen displays a set of soft keys which corresponds to the function keys on the computer keyboard. To enter a program (for example, the Utility Program) at the top level or perform an operation within a program simply press the corresponding function key. One must exit from a mode before gaining access to new operations in a different mode or entering a new program from Top Level.)
    1. Enter the Utility program. [F8](note: you should see the lower left label change from Top Level to Utility 1, indicating that you are in the Utility program). Press More Keys [F7] (note: you should see the lower left label change from Utility 1 to Utility 2). Now press [F5] to enter the Configure System mode (note: you should see the lower left label change again to indicate you are now in the configuration system mode within the Utility program.)
    2. Check to ensure that the HP 1090 Liquid Chromatograph and Diode array Detector are enabled and plotter available appears at the bottom of the screen.
    3. [Quit] from the Configuration mode and [Exit] from the Utility program back to Top Level

 

B. Checking the baseline: (warming up the diode array detector)

1. Enter the [Data Acquisition] Program to gain access to the "PREPARE TO INJECT" form.

2. Press [Monitor] to enter the MONITOR display window.
  1. Turn on the diode array detector by pressing [DAD Lamp On] and warm up about 30 min. If trace in upper window goes outside window hit [Zero Plot]. Recalibrate after a few minutes by pressing [Balance].

4. While baseline is stabilizing [Exit] to the PREPARE TO INJECT form.

 

C. Priming the system: (getting the pump ready)
  1. In the "PREPARE TO INJECT" form: (use the tab key or the cursor (arrow) keys to move the highlighted field to the different items).
  2. Set Flow equal to 1ml [Return].

3. Set Solvent A [on], and Solvent B [off]. Type 0% for channel B solvent composition .
  1. Turn on the pump by hitting [More Keys] and [Pump On]. Notice that the left side of the form indicates the parameters you set and the right side indicates the actual flow rate.

5. Check the stability of the high and low pressures by pressing [Diagnose] to gain access to the DIAGNOSTIC form. Monitor the pressures. They should not change by more than 2-3 units. If they do, consult instructor.

6. [Exit] from the DIAGNOSTIC display and return to the PREPARE TO INJECT form. [Pump Off].

7. Set channel A to [off], and channel B to [on]. Type 100% for channel B. Repeat steps 4-6 to prime Channel B.

8. Set all channels to [on].

9. Set channel B to 50%. [Pump On], [Diagnose]

10. Monitor stability of pressure readings.

11. [Pump Off] [Exit] to Top Level

 

D. Setting the mobile phase composition and flow rate:

1. Enter [Data Acquisition] mode.

2. Type in File name for data. (Example: Initials, Date, Run) (MM01.1)

3. Type flow rate.

4. Set solvent system by typing proper percentage for channel B.
  1. Set stop time by choosing [set by user] and type in stop time.
  2. If the pump has not yet been turned on, start it now. (See C. 4.)

 

E. Setting the detection parameters:

1. Press [Edit Param], [DAD params] to enter the DIODE ARRAY DETECTOR form .

2. Set Wavelength and Bandwidth for sample and reference. You may either use just one set of values or many. A typical choice might be 254 nm wavelength with 4 nm bandwidth for the sample and 400 nm wavelength and 100 nm bandwidth for the reference. If you choose more than one set of wavelengths, the chromatogram corresponding to the values selected in column A will be used in the default display. For Store Spectrum, use [next/previous choice] to choose "peak controlled". Enter desired Threshold and Peakwidth. Typical choices are 0.2 mAU for threshold and 0.16 min for peakwidth. Choose "same as LC" for Stop Time. Set spectrum range.

3. To delete a signal move cursor to Sample Wavelength and press [off].

4. [Exit] to the FILE INFORMATION form.

5. Delete any LC (liquid chromatograph) Time Tables left from previous analysis by hitting [LC Param], [LC Time Table], [More Keys], [Delete Table].

6. Press [Print Parms] to get a printout of your parameters.

7. If you wish to save the parameters press [Save Parameters]. Type the file name for the parameters. [Return]. This is necessary only if you want to be able to reload the same set of parameters at a later date. It is NOT necessary to save the parameters in order to just run a chromatogram.

 

F. Starting the run:

1. [Exit] to the PREPARE TO INJECT form. Check the data filename. Is it correct? (note: raw data is saved automatically under this filename once the run is started).

2. Move the lever at the injection port to load.

3. Inject the sample into port.
  1. Press [Monitor] if you wish to monitor the run. (note: red line indicates time zero at start of run). The horizontal and vertical scales of the monitor window can be adjusted with Time Window and Range, respectively. These do not affect the stored data, just the display.
  2. Pull lever on the injection port to the inject position to start the run. (note: lower right corner displays elapsed run time).

 

G. Obtaining a Spectrum:
  1. [Exit] to Top Level. Enter [Data Editor] Program.
  2. Type [?] to get a directory. [Select] file.
  3. Enter the Spec 1 mode by hitting [Spectra Keys] [New Spectrum]
    1. Use mouse or arrow (cursor) keys to position arrow at top of peak. (Type [1] to slow down cursor movement) [Return] .
    2. To obtain screen with spectra hit [More Keys] [Change Display]

 

H. Printing out data:
  1. To get a printout of chromatograph or spectrum [Exit] to Data Editor.
  2. [Graphics Keys] to enter graphics mode.
  3. If you wish to add labels or change the title:

a. [Annotate] Type text if you wish to add labels or change title.

b. [Position Label] [Return] [Write Label] [Exit]

4. [Dump Graphics] will print out on printer what appears on the screen.

 

I. Integrating data:

1. [Exit] to Data Editor program.

3. To integrate data press [Integ Keys], [Integ]

4. To set threshold press [Event Keys], [Initial Values], [Initial Threshold] Type value and [Return].

5. To get a listing of the integration methods press [List Events] [Send to Printer].

6. [Exit] back to Integration mode and press [Integ] to integrate.

7. To obtain a list of integrated peaks, [List Int Results] [Send to Printer]

8. To save integrated data, [Exit ] to Integration mode, press [Open Int File]. Type title. [Return]

 

J. Shutting down:
  1. Turn off the pump and the helium flow. If the system will not be used again soon, turn off the hardware in the reverse order from the initial power up procedure.

 

K. More Information:

The Hewlett Packard Manual entitled "HPLC ChemStation:Running An Analysis" is very complete, and will walk you through a lot of useful procedures.