Optimization of the isocratic non-aqueous reverse phase (NARP) HPLC separation oftrans/cis-α-and β-carotenes
✍ Scribed by Lesellier, E. ;Marty, C. ;Berset, C. ;Tchapla, A.
- Publisher
- John Wiley and Sons
- Year
- 1989
- Tongue
- English
- Weight
- 696 KB
- Volume
- 12
- Category
- Article
- ISSN
- 0935-6304
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✦ Synopsis
Non-aqueous reverse phase chromatography was used to optimize the separation of all-trans-a-and p-carotenes and their cis isomers extracted from dried carrots.Thetechnique can easily be applied to routine assays. Optimization was carried out by modifying the concentrations of methylene chloride and acetonitrile, and by studying the influence of temperature on the capacity factors and selectivity. The effect of the type of C 18 stationary phase bonding (mono-and polyfunctionalf on separation was also examined.The need for polymeric bonding on the stationary In adsorption liquid chromatography, a-and p-carotenes have been separated by semi-preparative chromatography on silica 191, whereas isomers of p-carotenes have been fractionated on alumina [lo] and on calcium hydroxide [4,11,12]. The resolution power of the latter adsorbent, however, is insufficient for the separation of a-carotene stereoisomers. Furthermore, results obtained with adsorption HPLC depend on a strict control ofthe water content of the system and column equilibration lasting several hours. These constraints render the technique ill suited for routine analyses.
. -phase is stressed for the realization of the trans/cis isomer separation.
The use of reverse phase partition HPLC has fewer constraints. It has been used to separate a-and p-carotenes (cis/trans isomers together) on a column of C18 bonded silica [13]. In general, the analysis of bulky hydrocarbon solutes in water/