Background: Esculin (ESCN) is used in the pharmaceutical industry with intravenous effect,
stimulant and anti-inflammatory capillaries, like vitamin P. It is a significant component of many
anti-inflammatory remedies such as esqusan, esflazid and anavenol . It is also found in numerous
other remedies available in the market such as proctosone, anustat, and ariproct.
Objective: To determine experimental conditions, to elucidate retention behavior of esculin in HILIC
mode. Moreover, to suggest new ways to separate and determinate esculin in ointments.
Methods: Two hydrophilic columns were obtained by attaching molecules of sulfobetaine to polystyrene-
divinylbenzene particles were studied for chromatographic separation of esculin. The various
lengths of the chain are used as an investigative instrument for esculin retention conduct. The change
of ACN percentage, buffer concentrations and pH of mobile phase in order to study the retention conduct
Results: A mixed mode hydrophilic interaction and ion exchange is the separation mechanism of esculin.
A calibration graph was created for two columns. The concentration range was 8-1200 ng.ml-1,
LOD 2.33 and 1.40, RSD% 0.31-1.02, , LOQ 7.07 and 4.25 ng.ml-1, Erel% 0.83 ± 0.68 and 0.545 ±
0.45, Recovery% 100.86 ± 0.68 and 1054 ± 0.45.
Conclusion: The findings of the current study introduced new ZIC-HILIC methods for the separation
and quantitation of esculin.