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HPLC Profiling with At-line Microdilution Assay for the Early Identification of Anti-fungal Compounds in Plants from French Polynesia

Published inPhytochemical analysis, vol. 25, no. 2, p. 106-112
Publication date2014
Abstract

Introduction – The search for anti-fungal compounds has maintained a scientific interest notably due to existing difficulties in the treatment of mycoses and their increasing occurrence in hospitals. Objective – Development of a simple method to rapidly identify anti-fungal compounds in crude plant extracts based on a HPLC microfractionation approach combined with an at-line anti-Candida assay. Methods – The scale of the semi-preparative HPLC microfractionation was adapted to fit the sensitivity of the Candida albicans anti-fungal in a 96-well microdilution assay. This format is also compatible for MS and NMR dereplication of the active compounds. Results – Based on the screening of 12 crude extracts of plants from French Polynesia, three plants, which displayed various levels of anti-fungal activities, were selected to assess the efficiency of the HPLC anti-fungal profiling and the scale necessary for microfractionation. The same anti-Candida assay was performed on the HPLC microfractions collected using a generic profiling method. Analysis of active microfractions by MS and NMR issued from the most active extract enabled an efficient dereplication of the compounds responsible for the anti-fungal activity. Conclusion – A generic HPLC anti-fungal profiling method was developed which revealed that only 50 mg of crude extract were sufficient for a rapid identification of compound(s) responsible for the anti-Candida activity. This approach was illustrated by the study of Alphitonia zizyphoides, a plant traditionally used to treat dermatomycoses.

Keywords
  • Microfractionation
  • Anti-fungal
  • Betulinic acid
  • Alphitonia zizyphoides
  • Candida albicans
Citation (ISO format)
BERTRAND, Samuel et al. HPLC Profiling with At-line Microdilution Assay for the Early Identification of Anti-fungal Compounds in Plants from French Polynesia. In: Phytochemical analysis, 2014, vol. 25, n° 2, p. 106–112. doi: 10.1002/pca.2473
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ISSN of the journal0958-0344
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