« Removing the column »
1. When the pump is running, push [pump] on the pump.
2. Open the cover of the column oven.
3. Remove the tube connecting to the inlet/outlet of the column.
2. Open the cover of the column oven.
3. Remove the tube connecting to the inlet/outlet of the column.
« Attaching the column »
1. Attach the column
NOTE: The fluid in the column inside the oven runs from the bottom up. Attach so that the column inlet is on the bottom.
1) Connect the SUS tubing out of PPSQ inside the column oven to the column-inlet.
2) Connect the tube out of the detector in the column oven to the column-outlet.
3) Close the cover of the column oven.
2) Connect the tube out of the detector in the column oven to the column-outlet.
3) Close the cover of the column oven.
2. Perform LC control.
1) In the [PPSQ Analysis] window, select [Sensoer Status] on the [Monitor] menu.
The [Status] window is displayed.
1) In the [PPSQ Analysis] window, select [Sensoer Status] on the [Monitor] menu.
The [Status] window is displayed.
2) In the [PPSQ Analysis] window, select [LC Control] on the [Maintenance] menu.
The [LC Control] dialog box is displayed.
3) Configure each function to the following value.
Pump: A.Flow 0.5
Detector: Wavelength(Starting point) 265, Wavelength(Ending point) 273, Lamp ON
Cplumn oven: Temp. 40, Control Heater ON
HINT: Usually,the method file used for the last analysis is displayed in the [File name] fied.
4) Remove the tube connecterd to the detector outlet from the mobile phase bottle and place it in a suitable container.
5) Click [Download] and [Activate]. PPSQ runs following the configured condition.
Confirm that fluid is draining from the tip of the detector outlet tubing.
6) For the isocratic system, when the column is full of mobile phase (i.e., when the container holding the end of the tube contains 20 to 30 mL of mobile phase), insert the tube connected to the detector outlet in the mobile phase bottle.
7) Seal the opening of the mobile phase bottle with parafilm or the like.
8) After the temperature of Column oven reaches at 40ºC, enter [1] on [A.Flow] of Pump and click [Download].
9) Click [Close] at [LC Control] window.
5) Click [Download] and [Activate]. PPSQ runs following the configured condition.
Confirm that fluid is draining from the tip of the detector outlet tubing.
6) For the isocratic system, when the column is full of mobile phase (i.e., when the container holding the end of the tube contains 20 to 30 mL of mobile phase), insert the tube connected to the detector outlet in the mobile phase bottle.
7) Seal the opening of the mobile phase bottle with parafilm or the like.
8) After the temperature of Column oven reaches at 40ºC, enter [1] on [A.Flow] of Pump and click [Download].
9) Click [Close] at [LC Control] window.
10) In the [PPSQ Analysis] window, select [Sensor Status] on the [Monitor] menu. Check the column pressure, the pump flow rate, and the column temperature by the [HPLC] tab in the [Status] window. The column pressure is normally in the range 8 to 11MPa at 40ºC by the flow rate of 1mL/min.
3. Checking the Baseline
1) Click [Chromatogram] icon in the [PPSQ Analysis] window.
HINT: Alternatively, select [LC Analysis] on the [Monitor] menu in the [PPSQ Analysis] window.
1) Click [Chromatogram] icon in the [PPSQ Analysis] window.
HINT: Alternatively, select [LC Analysis] on the [Monitor] menu in the [PPSQ Analysis] window.
2) If the baseline deviates from "0" on the vertical axis, press the [ZERO] button on the detector.
3) Check that the baseline is stable. (Baseline fluctuation in a 10-minute period is 2.5 mAU or less.)
3) Check that the baseline is stable. (Baseline fluctuation in a 10-minute period is 2.5 mAU or less.)
4. Inspecting for Leaks
1) Sometimes, immediately after system startup, inspect the piping connections (in particular, the column) to check that there are no leaks.
2) Analyze the PTH-amino acids mixed standard, check the status of the HPLC analysis system, and correct (or create) the quantitative parameters ID table as required.
1) Sometimes, immediately after system startup, inspect the piping connections (in particular, the column) to check that there are no leaks.
2) Analyze the PTH-amino acids mixed standard, check the status of the HPLC analysis system, and correct (or create) the quantitative parameters ID table as required.
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