P-Carotenes extraction

Dunaliella natural P-carotene is distributed widely in many different markets under three categories p-carotene extracts, Dunaliella powder for human use, dried Dunaliella for feed use. Extracted purified P-carotene is sold mostly in vegetable oil in bulk concentrations from 1 to 20% to color various food products and for personal use in soft gels usually containing 5 mg P-carotene per gel. Purified natural p-carotene is generally accompanied by the other Dunaliella carotenoids, primarily lutein, neoxanthin, zeaxan-thin, violaxanthin, cryptoxanthin, and a-carotene for a total of approximately 15% of carotene concentration. This compound is marketed as carotenoids mix. ... 

Solvent extraction removes chlorophyll and other pigments to give a light-colored product but increases processing costs. Furthermore, solvent extraction removes p-carotene and reduces vitamin A activity (89) (see Terpenoids Vitamins). Supercritical CO2 extraction at 30 and 70 MPa (4,350 and 10,150 psi) and 40°C removed 90 and 70% carotene and lutein, respectively, from alfalfa LPC (96). This process avoids organic solvent residues and recovers valuable by-products. 

Most of this amount is in the form of fucoxanthin in various algae and in the three main carotenoids of green leaves lutein, violaxanthin, and neoxanthin. Others produced in much smaller amounts but found widely are p-carotene and zeaxanthin. The other pigments found in certain plants are lycopene and capsanthin (Figure 2.2.1). Colorant preparations have been made from all of these compounds and obviously the composition of a colorant extract reflects the profile of the starting material. Carotenoids are probably the best known of the food colorants derived from natural sources. ... 

Nagao, A. et al., Stoichiometric conversion of all trans-P-carotene to retinal by pig intestinal extract, Arch. Biochem. Biophys., 328, 57, 1996. 

Because most food matrices are water soluble, many efforts were directed to the formulation of lipophilic pigments (mainly carotenoids) into water-soluble formulations (powders or gels). For hydrophilic pigments like flavonoids, polar dried microcapsules are the most popular ways to stabilize their functionality. Extracts rich in P-carotene were encapsulated using three different encapsulation techniques (spray drying, drum drying, and freeze drying)." ... 

Hejazi, M.A., Kleinegris, D., and Wijffels, R.H., Mechanism of extraction of P-carotene from microalga Dunaliellea salina in two-phase bioreactors, Biotechnol. Bioeng., 88, 593, 2004. 

An interlaboratory study using mixed vegetable reference material showed average relative standard deviations (RSDs) of 23% ranging from 11% for lutein and a-carotene to 40% for lycopene." Triplicate HPLC injections of the same extract showed RSD values of 0% for P-carotene and 6.8% for lutein. ... 

Chromatography may be necessary to separate some carotenoids. A saponified carotenoid extract from carrots was applied to OCC on MgO and two fractions were eluted. Successive crystalhzation with petroleum ether and MeOH was carried out to obtain a- and P-carotene crystals, with 99 and 98% purity levels, respectively. ... 

Considering the concerns of consumers for synthetic colorants and interest in natural formulas, many food manufacturers seek alternative healthy solutions to replace colorants, even the regulated ones from positive lists (like p-carotene), with colored fruit and vegetable extracts to be used as functional food ingredients or nutraceuticals (food supplements). ... 

Monarch epidermis. Peaks seen at 8.7, 10, and 82min are 3-hydroxy-10 -apo-P-carotenal, lutein, zeaxanthin, and P-carotene, respectively. The peak seen eluting at 22 min is the internal standard, monopropyl lutein ether, (b) The chromatogram obtained from an extract of the leaves of the milkweed plant. Peaks eluting prior to lutein are xanthophylls and epoxy xanthophylls, identified components include lutein, zeaxanthin, P-carotene, and its crT-isomer, eluting at 10, 11, 41, 77, and 79min, respectively. 

The natural dye was extracted by immersion of fresh Morns nigra (black mulberry) in ethanol for several hours. The pure violet dye extract, a blend of p-carotene and Morus nigra, and a composite blend of chlorophyll A and B, carminic acid, trans-P-carotene, and Morus nigra extracts (hereafter called Mix) were deposited on Ti02. 

Fig. 2.24. C30 chromatograms of carotenoids extracted from human serum (a) xanthophylls fraction, 7 93 (v/v) MTBE-methanol mobile phase (b) a- and / -carotenes fraction, 11 89 (v/v) MTBE-methanol mobile phase (c) lycopene fraction, 38 62 (v/v) MTBE-methanol mobile phase. Tentative peak identifications (a) 1, 13-c/s-lu- lutein 2, 13 r/.vlutein 3, a//-/ra s-lutein 4, zeaan-thin 5-7, unidentified P,e-carotenoids and 8, / -cyrptoanthin (b) 1-2, unidentified ae-carotene isomers 3, 15-eH -/f-carotenc 4, 13-cw-/ -carotene 5, all-trans-a-carotene 6, all-trans-P-carotene and 7, 9-ci.v-/3-carotene and (c) 1-11 and 13, c/s-lycopene isomers and 12, all-trans-lycopene. Reprinted with permission from C. Emenhiser el al. [51].

Fig. 8.3 UPLC analysis of Cupi/cMm-lyoplrilizEd pericarp carotenoids. Carotenoids detected by absorption at 454 nm, following separation on a waters acquity C18 1.8 xm HSS particle, 2.1 x 100 mm column resolved with 10% isopropanol (v/v) (a) and 100% acetonitrile (b). The solvent profile included two linear phases (0-3 min at 75% (b) 3-11 min from 95 to 100%) flow rate of 0.75 mL/min. (a) Standards (each at 100 ppm) capsorubin (1.7 min), capsanthin (2.08 min), antherxanthm (2.69 min), zeaxanthin (2.97 min), f -cryptoxanthin (4.86 min), and P-carotene (8.15 min), (b) Valencia pericarp extract, (c) NuMex Sunset pericarp extract...

Ye, L. Landen, W. O. Eitenmiller, R. R. Liquid Chromatographic Analysis of All-trans-Retinyf Pafmitate, P-Carotene, and Vitamin E in Fortified Foods and the Extraction of Encapsulated and Nonencapsulated Retinyl Pafmitate. J. Agric. Food Chem. 2000, 48, 4003-4008. 

The solubility of P-carotene in supercritical fluids has been studied extensively [81 to 85], The extraction of P-carotene from a wide varieties of natural sources has also been described like alfalfa-leaf protein concentrates [86], carrots [34,87], sweet potatoes [88], and algae [89],... 

If the extract or fraction is composed of one predominant carotenoid, it can be monitored at the appropriate wavelength and the A1 % for that carotenoid used. If the extract is composed of more than one carotenoid, the absorbance at 450 nm can be measured and the A1% for P-carotene used. Results can be expressed as total P-carotene equivalents. Alternatively a typical Al% value of 2500 can be used for comparing relative quantities between various sample extracts. An A1% value of 2500 is appropriate since the predominate carotenoids, with the exception of lycopene (Al% = 3450), have X between 2460 and 2710. Of course if samples of tomatoes are to be analyzed, which contain lycopene predominantly, then an A1% value of 3450 would be more appropriate. 

The complete separation from retinol to P-carotene requires -15 min. Figure F2.3.2 illustrates the separation of vitamins and carotenoids in the mixed food extract using this LC system. The elution order using this method is lutein, zeaxanthin, -cryptoxanthin, lycopene, a-carotene, and P-carotene. 

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