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Updated: Aug 21, 2026

Automated HPLC Separation Using LC-Mate: An Integrated Repetitive Autosampler and Fraction Collector for Microscale Purification
Published on: February 27, 2026
Improved separation method for highly purified lutein from Chlorella powder using jet mill and flash column
Shinya Shibata1, Chiyoko Ishihara, Keisuke Matsumoto
1Yakult Central Institute for Microbiological Research, 1796 Yaho, Kunitachi, Tokyo 186-8650, Japan. shinya-shibata@yakult.co.jp
Abstract:
We investigated an improved method for the separation of high-purified lutein from a commercially available spray-dried Chlorella powder (CP) using fine grinding by jet mill and flash column chromatography on a silica gel. Saponification and extraction of lutein were enhanced 2.3-2.9-fold in jet mill-treated CP (mean particle size, 20 microm) as compared to untreated CP (mean particle size, 67 microm). The carotenoid extract was dissolved in ether-hexane (1:1 v/v) and subjected to flash column chromatography on silica gel. A mixture of alpha- and beta-carotene was eluted with hexane, followed by elution with hexane-acetone-chloroform (7:2:1 v/v). Lutein (dark-orange band) was collected after the elution of an unknown colorless compound (detected based on UV absorbance). The purity of lutein in this fraction was over 99%, and the yield was 60%. The present study provides key information for obtaining highly purified lutein using flash column chromatography on a silica gel.
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