Preparations before quantitation

Description

The table below describes the preparations before the quantitation.

Step

Action

1

Create a method to be used for all the standard runs. The method and the injection volume must be the same for all the runs.

2

Perform at least one run for each concentration level of the standard.

3

Peak integrate the curves to produce a peak table for each of the standard curves.

Note: When integrated, all standards must use the same X-axis base unit. Time is the recommended unit for the highest reliability.

4

Check that each integration is correct and consistent.

5

Select File:Save to save all the peak tables.

Concentration levels

The standard series should include standard concentrations that extend beyond the lower and upper limits of the sample amount. If an internal standard is used, the internal standard must be added in the same concentration in all standards.

Methods created from a wizard

If the method is created from a wizard for ÄKTAdesign systems, you may select the correct standard concentration level in the variable Quantitation_Type. You can also set the level after the run has been performed. Each level is an alias for a specific concentration of the standard.

The list below describes how the levels are applied:

  • Level 1 should be selected for the standard with the highest or lowest concentration.

  • The levels must be set in consecutive order of changing concentration of the standard.

  • All runs with the same concentration must be given the same level.

Reject irrelevant peaks

If many small irrelevant peaks are detected, it may be an advantage to re-integrate after adjusting the Reject peaks criteria. The number of largest peaks to detect has a default value of 20 and it may be helpful if this is set to a smaller value.

2005-06-15