Related Experiment Video
Updated: Aug 29, 2026

Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification
Published on: February 27, 2026
Application of preparative high-speed counter-current chromatography for separation of chlorogenic acid from Flos
Hai-Tao Lu1, Yue Jiang, Feng Chen
1Department of Biology, Hong Kong Baptist University, Kowloon Tong, Hong Kong, China.
Abstract:
Chlorogenic acid, an ester formed between caffeic acid and quinic acid, is a major phenolic compound in the traditional Chinese medicinal herb Flos Lonicerae. The separation and purification of chlorogenic acid from the crude extract of Flos Lonicerae was achieved by high-speed counter-current chromatography (HSCCC). A high acid, highly polar two-phase solvent system containing n-butanol-acetic acid-water (4:1:5) was run on a preparative scale. The upper phase was used as the mobile phase in the head to tail elution mode. A 300-mg quantity of the crude extract containing 5.97% chlorogenic acid was loaded on a 342-ml HSCCC column. Double separations were performed with the same solvent system yielding 16.9 mg chlorogenic acid at 94.8% purity with approximately 90% recovery.
Related Concept Videos
High-Performance Liquid Chromatography: Introduction
In HPLC, two phases play a critical role in the separation process:
High-Performance Liquid Chromatography: Elution Process
Supercritical Fluid Chromatography
SFC utilizes a supercritical fluid mobile phase,...
High-Performance Liquid Chromatography: Instrumentation
Principles Of Column Chromatography
Chromatography: Introduction
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
