Scale-up purification for rutin hyrdrolysates by high-performance counter-current chromatography coupled with semi-preparative high-performance liquid chromatography

ABSTRACT

The study made the isolation of three constituents from rutin and exemplified how to achieve quercetin separation and quantification in rapid and scale-up processes by high-performance counter-current chromatography (HPCCC). Meanwhile, we also isolated the other two constituents, kaempferol and isorhamnetin, by semi-preparative high-performance liquid chromatography. After systematic optimization of solvent systems, sample concentration and flow rate on analytical Mini-DE centrifuge, the same conditions were scale-up by 50-fold in preparative Midi-DE centrifuge to purify quercetin, and the maximum sample loading achieved 1.65 g. Eventually, we successfully isolated quercetin, kaempferol and isorhamnetin with the purity of 99.80%, 99.84% and 99.95%, respectively. The process, therefore, provided an efficient method of obtaining sufficient quantities of highly purified quercetin, as well as kaempferol and isorhamnetin.

KEYWORDS:

• High-performance counter-current chromatography (HPCCC)
• isorhamnetin
• kaempferol
• quercetin
• rutin
• semi-preparative HPLC

Acknowledgements

Jinying Chen and Linhong He contributed equally to this work.

Funding

We are grateful to the National Key Programs of China during the 12^th Five-Year Plan Period (grant 2012ZX09103101-033).

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Additional information

Funding

We are grateful to the National Key Programs of China during the 12^th Five-Year Plan Period (grant 2012ZX09103101-033).