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Title: Fractionation of secondary metabolites of orange (Citrus sinensis L.) leaves by fast centrifugal partition chromatography


Submitted to: Proceedings Florida State Horticultural Science
Publication Type: Proceedings
Publication Acceptance Date: 12/5/2010
Publication Date: 12/29/2010
Citation: Manthey, J.A. 2010. Fractionation of secondary metabolites of orange (Citrus sinensis L.) leaves by fast centrifugal partition chromatography. Proceedings Florida State Horticultural Science. 123:248-251.

Interpretive Summary: Conventional methods of analysis of compounds in orange leaves are unable to resolve most of the minor compounds. This presents a major hurdle for researchers looking for compounds that can be used to detect diseases in plants. This work showed that by combining a new chromatography method (FCPC) with state of the art mass spectrometry, many of the heretofore undetected compounds can be analyzed.

Technical Abstract: Conventional HPLC provides ready detection of the major phenolic compounds in methanol extracts of orange leaves, yet conventional HPLC also shows the presence of many more compounds, to an extent where extensive peak overlap prevents distinct peak detection and reliable quantitation. A more complete chromatographic analysis of the complex compound profiles of orange (Citrus sinensis) leaves was made possible with separations by Fast Centrifugal Partition Chromatography prior to HPLC analysis. Separations by Fast Centrifugal Partition Chromatography were achieved using the biphasic system of ethanol/butanol/water (20/80/100, v/v/v). Early eluted fractions contained numerous polar and mid-polar hydroxycinnamates. Later eluting fractions contained various profiles of polar and mid-polar flavonoids, many of which significantly overlapped in the HPLC of the original orange leaf extract. Analysis of the Fast Centrifugal Partition Chromatography fractions by HPLC-time-of-flight mass spectrometry provided detection of many more compounds compared to other conventional methods of detection used with HPLC.