Abstract

The use of mobile phase additives enhances the separation and resolution of the bioactive compounds on the C18 column. Chlorogenic acid, caffeic acid, rutin, and scoparone from Herba Artemisiae Scopariae were investigated as the target compounds. Acetic acid, triethylamine, inorganic salts, and several ionic liquids were added as mobile phase additives into methanol/water (40:60, v/v). The result revealed that a mobile phase with 0.01 mol/L of ionic liquid [BMIM][BF4] enabled the optimum separation of the four target compounds.

Highlights

  • Herba Artemisiae Scopariae (HAS) is one of the oldest medicinal herbs in traditional Chinese medicine [1]

  • The majority of separations are impossible without various mobile phase additives, such as acids, salts, and organic compounds

  • The interactions between target compounds and the stationary phase were reduced with increasing concentrations of acetic acid in the mobile phase

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Summary

Introduction

Herba Artemisiae Scopariae (HAS) is one of the oldest medicinal herbs in traditional Chinese medicine [1]. Chlorogenic acid, caffeic acid, scoparone and rutin (the structures are shown in Figure 1) are the four major active components of the herb. Methanol, was used as the most common organic solvent in the mobile phase [8,9]. The majority of separations are impossible without various mobile phase additives, such as acids, salts, and organic compounds. There is no research about the effect of the mobile phase additives for the simultaneous separation of these four compounds. Methanol/water (40:60, v/v) was fixed as the mobile phase Several additives, such as acetic acid, triethylamine, and ionic liquids (1-butyl-3-methylimidazolium tetrafluoroborate ([BMIM][BF4]), 1-hexyl-3-methylimidazolium tetrafluoroborate ([HMIM][BF4]), 1-methyl-3-octylimidazolium tetrafluoroborate ([OMIM][BF4]), 1-ethyl-3-methylimidazolium methlsulfate ([EMIM][MS]), and. 1-butyl-3-methylimidazolium chloride ([BMIM][Cl]), were evaluated as mobile phase additives (Figure 2) for the separation of the mixture of the four compounds from HAS by RP-HPLC. The effects of the type and concentration of additives on the chromatographic performance are discussed

Retention Factor Estimation
Effect of Acid and Basic Additives on the Resolution
Effect of the Concentration of Inorganic Salt
Different Cations of Ionic Liquid
Effect of Anion of Ionic Liquid
Materials and Reagents
Preparation of Mobile Phases and Standard Solutions
Apparatus and Chromatographic Conditions
Conclusions
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