A rapid high-performance liquid chromatography method to measure lipophilic antioxidants in stressed plants: simultaneous determination of carotenoids and tocopherols
✍ Scribed by José I. García-Plazaola; José M. Becerril
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 128 KB
- Volume
- 10
- Category
- Article
- ISSN
- 0958-0344
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✦ Synopsis
A new reversed-phase HPLC method has been developed for the simultaneous determination of all major photosynthetic pigments in higher plants (antheraxanthin, aand b-carotene, chlorophyll a and b, lactucaxanthin, lutein, lutein-epoxide, neoxanthin, pheophytin a, violaxanthin and zeaxanthin) and tocopherols (aand g-tocopherol) in the same extract. The method is also suitable for the determination of other carotenoids and their intermediates, for example, gand z-carotene, lycopene, phytoene and phytofluene. The quantitative analysis of such compounds is of paramount importance in studies dealing with pigment biosynthesis and degradation, as well as in studies on plant ecophysiology, since many of these compounds seem to be important in enabling a plant to withstand environmental stresses. The method uses a C-18 column and a simple solvent gradient, resolving in 16 min all carotenoids and tocopherols. Chromatograms are generated by a photodiode array detector at three wavelengths (295, 410 and 445 nm). The use of a fluorescence detector significantly improves the sensitivity of detection of tocopherols and also allows the determination of d-tocopherol. The HPLC method provides a sensitive, accurate and reproducible procedure for the quantitative analysis of pigments and tocopherols and, since these compounds are usually analysed by separate analytical procedures in plant stress physiology studies, also represents a reduction in the requirements of equipment, the amounts of solvents used and the analysis time compared with the methods currently employed.