Method adjustment

Organic phase, pH value, injection volume, column temperature, etc.

Injection volume

Mobile phase: Methanol: 10mmol / L sodium hexanesulfonate (pH 2.8 buffer salt) = 35:65

Molar concentration ratio of aniline
to ion pair reagent
Plate noAsymmetryPeak type
1:8166891.05Normal
1:4137861.13Normal
1:2101671.50Slightly frontier
1:165272.01Serious frontier
2:159312.21Bimodal

Conclusion

When designing the method, it is recommended that the concentration of ion pair reagent should be at least 4 times larger than that of the sample, and the limit ratio should be 1:1. If the concentration of sample is too large than that of ion pair reagent, double peaks will appear.

Column Temperature

The change of temperature will cause a significant change in the relative retention value.
Because the column temperature will change the amount of ion pair reagents adsorbed on the chromatographic column, the selectivity will be changed due to the difference of ion pair reagent concentration.

Column type

There will be some differences, but in ion pair chromatography, the column filler will be partially covered by ion pair reagent. Therefore, changing the column type is not the preferred method.

Buffer salt type

It will have a certain effect and have little effect on selectivity.

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