ChromeGate 3.3.2 Software Manual - KNAUER Advanced Scientific ...

20 Short guide ChromGate®, KNAUER Instrument Control
Generating and Using a Calibration Curve
A calibration curve is normally generated via a sequence, which is
discussed in the chapter Quantitation and Using Sequence Files below.
Method File Commands
A method file can be saved, retrieved and edited as desired in the same
way as with other Windows applications. As an example, if the Method
dialog box is open and you want to save a new method, select
File/Method/Save as and enter the desired name.
Fig. 14 Save Method File as… Window
Similarly, if you want to open a method, select File/Method/Open… and
enter the file name.
A printed report describing all aspects of a method can be obtained by
selecting File/Method/Print.
Quantification and Using Sequence Files
Overview
Calibration involves the generation of the relationship between the
concentration of a compound and the area of the corresponding peak in
the chromatogram. Once calibration is established, it can be used to
determine the concentration of the compound of interest in unknowns.
A single level calibration involves a single sample to calculate the
calibration constant; the curve is generated from that point and the origin.
A multi-level calibration uses several standards (e.g. 10 µg/ml, 20 µg/ml,
50 µg/ml, 100 µg/ml).
Setting up a calibration involves the following steps:
1. Run the chromatograms containing the standards and
save them
2. Identify the peaks from the standards
3. Generate a peak table
4. Generate the standard curve
The standards used to generate a calibration should be well defined and
should be in a matrix that is similar to that containing the samples that
you will analyze. The standards undergo exactly the same sample
preparation process that is used for real samples.