Calibration window

Analysis Procedure: Step 1 -> Step 2 -> Step 3 -> Step 4

  • In the main Clarity window, open the Instrument on which you want to test the functions of Sequence. In Clarity Demo it is Instrument 1. Do so by using the Instrument - Login to Instrument X command or by clicking on its icon. The Login Dialog will open.
  • Let User field filled with the pre-selected Administrator. In the Select Project: combo-box choose option DEMO1 and press the OK button.
  • Use the Calibration button in the Instrument window to open the Calibration window.

Note:

If you wish to use prepared demo calibration instead of creating a new one, in the Calibration window, open (via the File - Open...command) the calibration file DEMO1.CAL and test the functions of the Calibration window on it. In this case you can skip the following section and continue with the chapter "Linking the calibration to a chromatogram".

  • Use the New Calibration icon ① to create a new calibration file. Save the calibration under a name of your choosing.

Note:

The calibration can be saved either using the Save Calibration icon ②, File - Save, or File - Save As... command.

The Calibration window - loaded standard

  • Use the Calibration Options icon ③ and change the Display Mode (top right corner of the dialog) to ISTD as our example is calculated by internal standard, then press the OK button.

Now, the calibration standards need to be imported into the calibration. This is done in a series of successive steps:

  • Use the Open Standard icon (yellow) ④ to open the STD 1.PRM data file. The lower part of the Calibration window now displays the chromatogram of the standard.
  • Use the Add All icon (blue) ⑤ to add all identified peaks to the Calibration Summary Table. The Table appears in the Calibration window, ready to be completed as shown on Fig "The Calibration window - loaded standard".
  • As demonstrated in the Calibration Summary Table and the Chromatogram, individual peaks are now identified according to their retention times only. To label the peaks, click and edit the fields in the Compound Name column ⑥ to the names shown on Fig "The Calibration window - loaded standard".
  • You may also set the color of a specific peak in the Peak Color column, for example, try to set the ISTD peak color to yellow.

Note:

The changes made in calibration immediately affects the chromatograms that have this calibration linked (so it is not influences the opened standard as it is not have this calibration linked).

  • Fill the Amount column ⑦ with the concentration of the particular compounds. In this standard mixture, all compounds except for the peak number 6 (ISTD) have the amount of 0.4.
  • Peak number 6 is our internal standard. So in the Is ISTD column, change its type to ISTD1 ⑧ and then set its amount in the Amount column to 2.

The first calibration level is now completed. On the tabs of the individual compounds ⑩ (labeled according to the Compound Name field), graphs with a single-point linear calibration can be viewed.

  • Proceed to completing the other calibration levels: the operation is quite simple and straightforward
    • Use the Open Standard icon (yellow) ④ again to open another calibration standard named STD 2.PRM.
    • Make sure that Calibration ⑨ is selected and use the Add All icon (blue) ⑤ (response will be added to first empty Level).
    • In the Amount column, enter the value of 1.0 for all peaks, except for ISTD peak 6, where you should enter a value of 2 instead.
  • Set the third calibration level analogously using the STD 3.PRM file and the Amount of 3.0, and subsequently the fourth level (file STD 4.PRM, Amount 5.0). Again, the ISTD peak should always have the Amount of 2.

As the result, on the tabs of the individual compounds ⑩, the linear four-point calibration graphs can be viewed.

  • Now, save the calibration file using the Save Calibration icon.