Planta Med 2011; 77 - WSI1
DOI: 10.1055/s-0031-1282092

On-line coupling of Centrifugal Partition Chromatography (CPC) to High Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS/MS)

T Michel 1, E Destandau 1, C Elfakir 1
  • 1Institute of Organic and Analytical Chemistry (ICOA), University of Orléans-CNRS UMR 6005, BP 67059, 45067 Orléans cedex 2, France.

Phytochemical analyses of food and medicinal plant extracts require rapid screening and detection strategies to identify bioactive natural products. Centrifugal Partition Chromatography (CPC), a free solid support liquid-liquid chromatography, is a well established method for the isolation of natural products and fractionation of complex samples at the preparative scale [1,2]. Nevertheless, even if the separation is monitored by detectors the composition of the different collected fractions must be evaluated by further High Performance Liquid Chromatography (HPLC) which is time consuming and give delayed information on the fraction composition.

We present here the development of a versatile tool for fast screening and rapid detection of bioactive natural products from plant extracts: the on-line coupling of CPC-UV to HPLC-UV-MS/MS. The coupling of CPC and HPLC systems, via a six position switching valve, reduces the time of complete fractionation procedure by direct on-line analyses of collected fractions and permits a guided fractionation step [3]. HPLC columns suitable for fast analysis, monolith and fused core columns, were evaluated to allow rapid analyse of compounds separated by CPC. Furthermore, the use of MS in tandem mode allows to get a direct structural identification of separated molecules.

This strategy was applied to the fractionation and purification of bioactive compounds from berries and roots of sea buckthorn (Hippophaë rhamnoides L., Elaeagnaceae), a Eurasian medicinal thorny bush, known to have various pharmacological effects [4].

Keywords: On-line CPC-HPLC-MS/MS, fractionation, purification, Hippophaë rhamnoides

References: 1. Marston A, Hostettmann K (2006)J Chromatogr A 1112: 181–194

2. Ingkaninan K et al. (2000)J Liq Chromatogr Relat Technol 23: 2195–2208

3. Michel T et al. J Chromatogr A Available online 1 February 2011

4. Guliyev VB et al. (2004)J Chromatogr B 812: 291–307