How to analyze high performance liquid chromatography

Chromatogram, in fact, is simply a two-dimensional spectrum with time as abscissa and electric signal as ordinate.

In high performance liquid chromatography, you can simply imagine that the stationary phase is a porous sponge-like cylindrical structure, and the sample enters and exits from the hole. Because the adsorption capacity of each substance is different, the distance will be drawn in the chromatogram.

Parameters related to the experiment:

1, retention time-that is, qualitative data parameters.

If the same sample is analyzed by using the same chromatographic column and the same mobile phase, the retention time of the sample should be fixed. Chromatographic peaks with different retention times should show different substances. If you run reversed-phase chromatography, the farther the chromatographic peak is, the smaller the corresponding polarity is.

2. Peak area-that is, a quantitative data parameter.

This is a parameter that you can read in the chromatogram. Under the same chromatographic conditions, the concentration of the same substance is proportional to the peak area. That is to say, if you prepare 1.0mg/ml of substance X and the peak area after injection is 10000, then if you prepare 0.5mg/ml of substance X, the peak area after injection is almost 5000.

3. Wavelength-this is a prerequisite for the smooth progress of the experiment.

The same sample, the same method and the same chromatographic column have different peak areas at different wavelengths. A substance means that it absorbs certain special wavelengths. For example, a substance has absorption at 2 10nm and 254nm. The substance may not be detected at a wavelength of 280 nm. So an experimental method starts with wavelength scanning.