Carotenoids solid-phase extraction

Ultrasound-assisted extraction provides efficient extraction in a shorter processing time than is needed for conventional extraction. The aid of ultrasound will result in a higher extraction yield and reduce solvent consumption. The extract may exhibit a wide range of colors (pale yellow, brown, red). For anthocyanin extraction in an acidic environment, the extract will be deep red, pink, or purple. The extract may contain considerable amounts of lipophilic compounds (e.g., chlorophyll, carotenoids, lipids). Prior to solid-phase extraction, those compounds can be eliminated from the extracts using liquid-liquid extraction. 

The removal of sterols, vitamin E vitamers, carotenoids, and other interfering material from the unsaponifiable fraction of food samples has been achieved using one or more of the following techniques coprecipitation of sterols with digitonin (91), precipitation of sterols from a methano-lic solution (195,209), adsorption chromatography on open columns of alumina (70,91,96), thin-layer chromatography on silica plates (209), and solid-phase extraction on silica (68,100) and reversed-phase (210) cartridges. 

After mixing the biological sample with the sorbent material, the homogenised blend is packed in a solid-phase extraction (SPE) column. After washing out polar interferences with water, the carotenoid stereoisomers are eluted in a concentrated fraction, requiring only small amounts of solvent, and thus leading to a complete and reproducible extraction. 

Figure 5.2.2 Scheme for the isolation and separation of geometrical carotenoid isomers from serum samples employing on-line solid phase extraction (SPE)-HPLC coupling... 

Extraction and cleanup procedures usually require solid-phase extraction based on commercially available Cig cartridges, for instance, after liquid extraction with common organic solvents (methylene dichloride, chloroform, acetonitrile).This step appears to be necessary to remove most of the interfering components such as carotenoids or chlorophyls, which are highly abundant in plants. Table 1 gives a list of some of the mycotoxins which can be analyzed by... 

High performance liquid chromatography (HPLC) has been by far the most important method for separating chlorophylls. Open column chromatography and thin layer chromatography are still used for clean-up procedures to isolate and separate carotenoids and other lipids from chlorophylls and for preparative applications, but both are losing importance for analytical purposes due to their low resolution and have been replaced by more effective techniques like solid phase, supercritical fluid extraction and counter current chromatography. The whole analysis should be as brief as possible, since each additional step is a potential source of epimers and allomers. 

Matrix solid-phase dispersion (MSPD) is the extraction method of choice for the analysis of solid samples, such as plant material, foodstuffs or tissue samples [26]. This method has been developed especially for solid or viscous matrices. MSPD is preferable to other extraction techniques, because the solid or viscous sample can be directly mixed with the sorbent material of the stationary phase [27]. As the carotenoid stereoisomers stay bound in their biological matrix until the elution step, they are protected against isomerisation and oxidation [28]. The extraction scheme of MSPD is shown in Figure 5.2.1. 

Figure 5.2.1 Scheme for the extraction of geometrical carotenoid isomers from solid biological samples employing matrix solid phase dispersion (MSPD)... 

A similar study has been carried out in order to test the capacity of RP-HPLC for the authenticity test of chilli powders on the basis of pigment composition. Carotenoid pigments were extracted by shaking 3 g of chilli powder with 10 ml of acetone for 30 min. The supernatant was decanted and the procedure was repeated as the solid rest was nearly colourless. The collected organic phases were evaporated and redissolved in the mobile phase. Separations were performed on a narrow-bore ODS column (150 X 2 mm i.d., carbon loading, 9.5 per cent). Eluents A and B were methanol-ACN (80 20, v/v) and bidistilled water, respectively. Gradient elution was initiated by 15 per cent A increased to 80 per cent A in 25 min, held for 10 min, increased to 90 per cent A in 10 min, held for 10 min, increased to 97 per cent A in 3 min and held for 62 min. Each step of gradient elution was linear. Measurements were... 

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