Planta Med 2008; 74 - PC81
DOI: 10.1055/s-0028-1084599

Use of FCPC (Fast Centrifugal Partition Chromatography) for the purification of a crude methanolic extract of the marine lichen Lichina pygmaea

C Roullier 1, M Chollet-Krugler 1, A Bernard 1, J Boustie 1
  • 1EA 4090 „Substances lichéniques et photoprotection“, UFR Sciences pharmaceutiques et biologiques, Université de Rennes1, 2 av. Pr. Léon Bernard, 35043 Rennes Cedex, France

Natural products crude extracts are often complex material, with several molecules, the separation of which commonly requires solid stationnary phase. Compared to these more traditional techniques, FCPC benefits from a number of advantages: no irreversible adsorption; total recovery of injected sample; tailing minimized; low risk of sample denaturation; low solvent consumption; favourable economics [1]. This technique is based on the partition between two immiscible phases of a solvent system. Our separative work focuses on a black marine lichen: Lichina pygmaea. This small fructicose lichen results from a symbiosis between a fungus (Ascomycete) and a cyanobacteria (Calothrix genus). It is poorly studied for its secondary metabolites, but has already been described to contain Mycosporine-Glycine as an antioxidant [2,3]. After HPLC identification of this molecule in the methanolic extract, we have developed a FCPC method to separate compounds, minimizing as much as possible subsequent purification. This crude extract of Lichina pygmaea is very complex, containing more than fifteen components, in a wide range of polarity (chlorophyll pigments, carotenoids, sugars, mycosporines). The solvent system we used was (butanol/acetic acid/water 4:1:5) as it provides a partition coefficient (Kd) between 0.5 and 1.5 for the targeted components. We used the multiple dual-mode as described by Delannay [4]. In one step, we have then easily separated a great number of molecules, without any adsorption problem, and with a few solvent consumption.

Acknowledgements: Association pour la Recherche sur le Cancer (ARC) (PhD grant for C. Roullier)

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2. De La Coba Luque, F. et al. (2007) WO/2007/0260363, Espagne.

3. Oren, A. et al (2007) FEMS Microbiol lett. 269: 1–10.

4. Delannay, E. et al. (2006)J. Chromatogr. A 1127: 45–51.