High-performance liquid chromatography pigment separation

FIGURE 7.12 High-performance liquid chromatography (HPLC) separation of pigments present in a green tissue. Peaks 1, neoxanthin 1, neoxanthin isomer 2, violaxanthin 2, violaxanthin isomer 3, luteoxanthin 4, anteraxanthin 5, lutein 5 and 5", lutein isomers 6, chlorophyll b 6, chlorophyll b C-132 epimer 7, chlorophyll a 7, chlorophyll a C-13 epimer 8, (i-carotene 8, cw-P-carotene isomer. 

FIGURE 7.13 High-performance liquid chromatography (HPLC) separation of a mixture of standards of possible pigments present in an extract of algal origin. Peaks 1, chlorophyUide a 2, chlorophyll c 3, pheophorbide a 4, fiicoxanthin 5, diadinoxanthin 6, lutein 7, chlorophyll h 8, chlorophyll a 9, asteroidenone 10, 3-carotene 11, pheophytin a. 

FIGURE 7.15 High-performance liquid chromatography (HPLC) separation of pigments present in canned peas. Peaks 1, pheophorbide b 1, pheophorbide b C-13 epimer 2, pheophorbide a 2, pheophorbide a C-13 epimer 3, neochrome 3, neochrome isomer 4, piropheophorbide b 5, piropheophorbide a 6, auroxanthin 6, auroxanthin isomer 7, muta-toxanthin 8, lutein 8 and 8", lutein isomers 9, pheophytin b 9, pheophytin b C-13 epimer 10, (3-carotene 11, pheophytin a 11, pheophytin a C-13 epimer 12, pyropheoph3din b 13, pyropheophytin a. 

Vanheukelem, L. et al., Improved separations of phytoplankton pigment using temperature-controlled high-performance liquid chromatography. Mar. Ecol. Prog. Ser., 14, 303, 1994. 

Saito, K., A new enzymatic method for extraction of precarthamin from dyer s saffron (Carthamus tinctorius) florets, Z. Lebensmitt. Untersuch. Forsch., 197, 34, 1993. Cserhati, T. et ah. Separation and quantitation of colour pigments of chili powder (Capsicum frutescens) by high-performance liquid chromatography-diode array detection, J. Chromatogr. A, 896, 69, 2000. 

T. Cserhati, E. Forgacs, H. Morais and C. Ramos, Use of a microbore ODS column for the separation of paprika (Capsicum annuum) pigments by high performance liquid chromatography. Pol. J. FoodNutr. Sci. 11/52 (2002) 11-13. 

However, during the past three decades, an analytical method has been developed that currently rivals and may soon surpass the traditional liquid chromatographic techniques in importance for analytical separations. This technique, high-performance liquid chromatography (HPLC), is ideally suited for the separation and identification of amino acids, carbohydrates, lipids, nucleic acids, proteins, pigments, steroids, pharmaceuticals, and many other biologically active molecules. 

Usha, T., Sarada, R., Ramachandra, Rao, S., and Ravishankar, G.A. 1999. Production of astaxanthin in Haematococcus pluvialis cultured in various media. Bioresource Technol. 68, 197-199. Vanheukelem, L., Lewitsu, J., Kata, T.M., and Craft, N.E. 1994. Improved separations of phytoplankton pigments using temperature controlled high performance liquid chromatography. Mar. Ecol. Prog. Ser. 114, 303-313. 

As indicated in our earlier article, the direct polymerization of monopyrroles to porphyrins is of little synthetic value as far as Ae unsymmetrical natural pigments are concerned. However, this might provide a feasible approach at l( ast to the copro- and uroporphyrins if sufficiently sophisticated methods could be developed for their chromatographic separation on a preparative basis. This has already been achieved analytically by high performance liquid chromatography (hplc). ... 

Braumann, T. and Grimme, L.H., Single-step separation and identification of photosynthetic pigments by high-performance liquid chromatography, J. Chromatogr., 170, 264, 1979. 

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