Detection of chlorophyll derivatives in soybean oil by HPLC

Abstract

Chlorophyll derivatives have been isolated from a degummed soybean oil by cellulose column chromatography and resolved by reversed phase HPLC. The HPLC separation was performed on a Zorbax ODS column using acetone‐methanol (75:25) as the mobile phase. Seven major components were detected by visible (650 nm) light absorption. Pheophytin A is the predominant component of the mixture (40–45% of the total). Pheophytin A’, pyropheophytin A, and three unidentified pigments having spectral features resembling pheophytin A comprise the other major pigments. No evidence was found for the presence of chlorophylls A and B in this oil.

Simultaneous Determination of Chlorophylls, Pheophytins, β‐Carotene, Tocopherols, and Tocotrienols in Olive and Soybean Oils by High‐performance Liquid Chromatography

“…The test tube was rinsed with acetone into a 10-mL volumetric flask and brought to volume with acetone. The prepared pheophytins were shown to be chromatographically pure, and the purity was further checked by comparing their visible spectra and molar extinction coefficients to those of pure pheo-phytins reported in the literature (Fraser and Frankl 1985). Dilutions of the standards, prepared with isopropanol-hexane (1.5:98.5 vol/vol), were stored under refrigeration and were protected from light during preparation and handling.…”

“…High-performance liquid chromatography (HPLC) is widely used for determining chlorophyll pigments, carotenoids, tocopherols, and tocotrienols in vegetable oils (Fraser and Frankl 1985;Csallany 1985, 1991;Mehlenbacher and others 1990;Endo and others 1992;Minguez-Mosquera and others 1992;Carvalho and others 1995;Psomiadou and Tsimidou 1998;Ye and others 1998). Most published procedures use one type of detector and rely on multiple injections to measure several different components because of differences in absorption and fluorescent properties between chlorophylls, carotenes, tocopherols, and tocotrienols.…”

Copper chlorophyll in olive oils: identification and determination by LIF capillary electrophoresis

“…It is more stable when exposed to light and heat, and more resistant to oxidative agents in general (Fraser & Frankl, 1985).…”

Rapid determination of chlorophylls in vegetable oils by laser‐based fluorometry

“…Since Chl is one of the many organic compounds which exhibit fluorescence, this property forms the basis of HPLC analysis using fluorescence detectors as well as other detectors (Eskins and Dutton 1979;Fraser and Frank1 1985;Rahmani and Csallany 1985;Jordan and Mudderman 1988;Aitzemuller 1989;Daun and Thorsteinson 1989;Davies et al 1990;Pfannkoch and Gill 1990). Chl remaining in refined edible oils are usually below the limit of detection by absorption spectrometry.…”