Simultaneous Separation and Purification of Microcystin LR, RR, YR by Using Supercritical Fluid Extraction and Column Chromatography

Abstract

An efficient method for the simultaneous separation and purification of microcystins (LR), (RR), and (YR) from the cyanobacterium is described. The method involves supercritical fluid extraction for the fast extraction and column chromatography for the purification of the microcystins LR, RR, and YR. A new method for the simultaneous extraction of microcystins LR, RR, and YR was developed by using supercritical CO₂. Three kinds of microcystins were successfully extracted with a ternary mixture (90% CO₂, 9.4% acetic acid, 0.6% water). The separation and purification procedure results in a purity of up to 95% for microcystins RR and YR without the need for a preparative high performance liquid chromatography (HPLC). Only for the purification of microcystin LR, we used a further preparative HPLC purification step. The advantage of the method developed here is that the sample handling steps were minimized, thus reducing possible losses of microcystins and saving extraction and purification time.

Keywords:

• Extraction
• Microcystin
• Purification
• Separation
• Simultaneous

Acknowledgment

This work was supported by grant no. R05‐2003‐000‐10157‐0 from the Basic Research Program of the Korea Science and Engineering Foundation.