Method adjustment

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

Injection volume

Mobile phase: methanol: 10mmol/L sodium hexanesulfonate (pH2.8 buffer salt) = 35:65

Molar concentration ratio of
aniline to ion-pairing reagent
Number of theoretical
Peak shape
1:2101671.50Slightly cutting edge
1:165272.01Serious frontier
2:159312.21Twin peaks

Conclusion: When designing the method, it is recommended that the ion-pairing reagent concentration is at least 4 times greater than the sample, and the limit ratio is 1:1. If the sample concentration is too much greater than the ion-pairing reagent concentration, double peaks will appear.

Column temperature

Changes in temperature can cause significant changes in relative retention.

Since the column temperature will change the amount of ion-pair reagents adsorbed on the chromatographic column, the selectivity will change due to the difference in the concentration of the ion-pair reagents.

Column type

There will be some variance, but since in ion-pair chromatography the column packing materials are partially covered by the ion-pair reagent, switching column types is not the preferred approach.

Buffer salt type

There will be a certain effect, with little impact on selectivity.

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