Isolation of Plastoquinone from Spinach by HPLCMarco Malferrari* and Francesco Francia
Laboratorio di Biochimica e Biofisica, Dipartimento di Farmacia e Biotecnologie, FaBiT, Università di Bologna, 40126 Bologna, Italy
- *Corresponding Author:
- Marco Malferrari
Laboratorio di Biochimica e Biofisica
Dipartimento di Farmacia e Biotecnologie
FaBiT, Università di Bologna, 40126 Bologna, Italy
Tel: +39 0512091293
Fax: +39 051 242576
E-mail: [email protected]
Received date: September 19, 2014; Accepted date: October 24, 2014; Published date: October 31, 2014
Citation: Malferrari M, Francia F (2014) Isolation of Plastoquinone from Spinach by HPLC. J Chromatogr Sep Tech 5:242. doi:10.4172/2157-7064.1000242
Copyright: © 2014 Malferrari M, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
We report a method for the purification of plastoquinone-9 (PQ), a prenylquinone cofactor involved in the photosynthetic electron transport chain. The described procedures relies on spinach-chloroplast isolation followed by PQ extraction and chromatographic fractionation. Extraction of PQ was achieved using partition of chloroplast suspension with methanol:petroleum ether. This procedure removed large amounts of green pigments from the extract and thus facilitates the subsequent chromatographic isolation of PQ. To obtain pure PQ, the developed extraction was combined with a two-step chromatographic approach using orthogonal stationary phases, i.e. alumina and octadecylsilane (C18). A small scale protocol for analytical reverse-phase high performance liquid chromatography (RP-HPLC), which may be implemented in most laboratories equipped with conventional systems, is described. The reported methodology represents a valuable tool for the fast production of small amounts of PQ, for which there are no commercial standards available.