Chromatography of chlorophylls

Schwartz, S.J., Woo, S.L., and Von Elbe, J.H., High-performance liquid chromatography of chlorophylls and their derivatives in fresh and processed spinach, J. Agric. Food Ghem., 29, 533, 1981. 

SCHOLZ B and ballschmiter k (1981), Preparation and reversed-phase high-performance liquid chromatography of chlorophylls , J Chromatogr, 208, 148-155. 

Suzuki, N., Saitoh, K., and Adachi, K. (1987). Reversed-phase high-performance thin-layer chromatography and column liquid chromatography of chlorophylls and their derivatives. J. Chromatogr. 408 181-190. 

M Zapata, JL Garrido. Influence of injection conditions in reversed-phase high performance liquid chromatography of chlorophylls and carotenoids. Chromatographia 31 589-594, 1990. 

Shioi, Y., Analytical chromatography of chlorophylls, in Chlorophylls, Scheer, H. (Ed.), CRC Press, Boca Raton, Ann Arbor Boston, London, 1991. 

Jeffrey, S.W., Quantitative thin-layer chromatography of chlorophylls and carotenoids from marine algae, Biochim. Biophys. Acta, 162, 271, 1968. 

Sahlberg, I. and Hynninen, P.H., Thin-layer chromatography of chlorophylls and their derivatives on sucrose layers, J. Chromatogr., 291, 331, 1984. 

Wright, S.W. and Shearer, J.D., Rapid extraction and high-performance liquid chromatography of chlorophylls and carotenoids from marine phytoplankton, J. Chromatogr., 294, 281, 1984. 

Yoshiura, M., Iriyama, K., and Shiraki, M., High-performance hquid chromatography of chlorophylls and some of their derivatives, Chem. Lett., 281, 1978. 

Chlorophyll b [519-62-0] M 907.52, sinters at 86-92 , sinters at 170 , dec at 160-170 , m 183-185 , 190-195 , [alj, -267 (Me2CO + McOH), [a] j-133 (McOH + Pyridine 95 5). See purification of chlorophyll a, and is separated from "a" by chromatography on sucrose [UV, IR Stoll and Weidemann Helv Chim Acta 42 679, 681 7959]. It forms red-black hexagonal bipyramids or four sided plates from dilute EtOH and has been recrystd from CHCl3-MeOH. It is soluble in MeOH, EtOH, EtOAc and insoluble in pet ether. [J Am Chem Soc 88 5037 1966.]... 

Buskov, S., Sprensen, H., and Sprensen, S., Separation of chlorophylls and their degradation products using packed column supercritical fluid chromatography (SEC), J. High Resol. Chromatogr, 22, 339, 1999. 

Watanahe, T. et al.. Preparation of chlorophylls and pheophytins hy isocratic liquid chromatography. Anal. Chem., 56, 251, 1984. 

Chapter 13 is devoted to the PLC of namral pigments, which encompass fla-vonoids, anthocyanins, carotenoids, chlorophylls and chlorophyll derivatives, porphyrins, quinones, and betalains. Chromatography of pigments is especially difficult because many are photo- and air-sensitive and can degrade rapidly unless precautions are taken. 

Boto and Bunt [465] used thin-layer chromatography for the preliminary separation of chlorophylls and phaeophytins from seawater, and combined this with selective excitation fluorometry for the determination of the separated chlorophylls a, b, and c, and their corresponding phaeophytin components. An advantage of the latter technique is that appropriate selection of excitation and emission wavelengths reduces the overlap among the emission spectra of the various pigments to a greater extent than is possible with broadband excitation and the use of relatively broadband filters for emission. 

Development of fast, accurate, and reproducible high-performance liquid chromatography (HPLC) methods has offset the use of traditional open-column and TLC methods in modern chlorophyll separation and analysis. A number of normal and reversed-phase methods have been developed for analysis of chlorophyll derivatives in food samples (unit F4.4), with octadecyl-bonded stationary phase (C]8) techniques predominating in the literature (Schwartz and Lorenzo, 1990). Inclusion of buffer salts such as ammonium acetate in the mobile phase is often useful, as this provides a proton equilibrium suitable for ionizable chlorophyllides and pheophorbides (Almela et al., 2000). 

Canjura, F.L. and Schwartz, S.J. 1991. Separation of chlorophyll compounds and their polar derivatives by high performance liquid chromatography. J. Agric. Food Chem. 39 1102-1105. 

During LC/MS, the glycerol matrix is included in the mobile phase and does not interfere with reversed-phase chromatography or in-line absorbance detection of chlorophylls. Matrices other than glycerol might be suitable for continuous-flow FAB of chlorophylls, but none have been reported. 

Harris, P.G., Carter, J.F., Head, R.N., Harris, R.P., Eglinton, G., and Maxwell, J.R. (1995) Identification of chlorophyll transformation products in zooplankton faecal pellets and marine sediment extracts by liquid chromatography/mass spectrometry atmospheric pressure chemical ionization. Rapid Cornmun. Mass Spectrom. 9, 1177-1183. 

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