A new chromatographic method utilizing the hydrogen bonding properties of Sephadex LH-20 gel in methanol / chloroform, was developed for the isolation of phenolic acids. This method was applied to the characterization of phenolic acids in flue-cured tobacco. Gel chromatography successfully isolated and concentrated the phenolic acids from other acidic substances and allowed identification and quantitation of these compounds by GC and GC-MS, as their trimethylsilyl derivatives. Compounds identified included: cis- and trans-isomers of p-coumaric, ferulic, caffeic, and sinapic acids; o-, m-, and p-hydroxybenzoic acids; o-hydroxyphenylacetic acid; 2,5- and 3,4-dihydroxybenzoic acids; and 2,3-, 2,5- and 3,4-dihydroxybenzaldehydes. Caffeic acid was the major compound, probably arising from the degradation of the chlorogenic acid of leaf. Aliphatic acids having two or more carboxyl groups were also isolated and included the following: malonic, succinic, fumaric, malic, and citric acids. A dihydroxycinnamaldehyde and a dihydroxynaphthoic acid are reported in tobacco leaf for the first time.
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1. StedmanR.L.: The chemical composition of tobacco and tobacco smoke; Chem. Rev. 68 (t968) t53-207.
2. Herrmann K.: Die phenolischen Inhaltsstoffe des Tabaks; Dtsch. Apoth.-Ztg. tot (t96t) t481-1482.
3. Herrmann K.: Ober die phenolischen Inhaltsstoffe des Tabaks und des Tabakrauches; Beitr. Tabakforsch. 2 (t964) 159-t79.
4. Andersen R. A. and G. Moegling: Quantitative gas chromatographic assay for caffeic acid moieties in tobacco; Anal. Biochem. 27 (t969) 397-408.
5. Yang C. and S. H. Wender: Free phenolic acids of cigarette smoke and tobacco paper chromatography Separation and identification; J. Chromatogr. 8 (1972) 82-89.
6. Bock F. G. A. P. Swain and R. L. Stedman: Composition studies on tobacco XLIV. Tumor-promoting activity of subfractions of the weak acid fraction of cigarette smoke condensate; J. Natl. Cancer Inst. 47 (1971) 429-436.
7. Trippett S. and D. M. Walker: The phosphobetaines Preparation and properties; J. Cbem. Soc. t962 1266-t272.
8. Bravo P. C. Ticozzi and A. Cezza: 1.1te reaction of carbonyl-stabilized phosphonium and arsonium ylides with o-hydroxybenzaldehydes Synthesis of a-hydroxy chalcones; Gazz. Chim. ltal. 105 (1975) 109-115.
9. Draffan G. H. R. N. Still well and J. A. McCoskey: ElC(;tron impact-induced rearrangement of trimethylsilyl groups in long chain compounds; Organic Mass Spec. 1 (1968) 669-685.
10. Morita H. J.: Identification of phenolic acids by gas chromatography- mass spectrometry; J. Chromatogr. 71 (1972) 149-153.
11. Horvat R. J. and S. D. Senter: A GLC method for analysis of phenolic acids in plants; J. Agric. Food Chem. 28 (t980) 1292-1295.