Keith R. Price

Analysis of the Major Flavonol Glycosides Present in Four Varieties of Onion (Allium cepa) and Changes in Composition Resulting from Autolysis

The major flavonoids of mature onion bulb were confirmed as the 3,4′-O-diglucoside (Qdg) and 4′-O-monoglucoside (Qmg) of quercetin using a combination of chromatographic comparisons, mass spectrometry and nuclear magnetic resonance spectroscopy. These two components account for over 85% of the total flavonoids in three varieties of onion with Qdg as the main component. Quercetin is detected in these long stored onions but only at low levels of less than 2% of the total. The remaining flavonoid fraction (approx 15%) comprises upto 17 different components of which quercetin-3-O-glucoside and isorhamnetin glucoside are prominent members although each contribute less than 1% of the total flavonoid fraction. There are significant differences in the levels of Qdg and Qmg between the four different onion varieties analysed; Qdg varying from 50–1300 mg kg-1 fresh onion tissue and Qmg from 36–394 mg kg-1. Maceration of the tissue for the three varieties tested led to a loss of Qdg and the appearance of Qmg and free quercetin. In the variety Rijnsburger 50% of the Qdg was degraded in 5 h and had completely disappeared after 24 h. These changes in Qdg can be quantitatively explained by increases in Qmg and free quercetin. The possible significance of quercetin glycosides in the diet is discussed.

Composition and content of flavonol glycosides in broccoli florets (Brassica olearacea) and their fate during cooking

The two main flavonol glycosides present in broccoli florets were identified as quercetin 3-O-sophoroside and kaempferol 3-O-sophoroside. Three minor glucosides of quercetin and kaempferol were also detected, namely isoquercitrin, kaempferol 3-O-glucoside and a kaempferol diglucoside. The sophorosides of quercetin and kaempferol were present in raw florets at a level of 65 mg kg−1 and 166 mg kg−1 fresh weight, respectively. The total content of quercetin and kaempferol glycosides expressed as aglycone was 43 and 94 μg g−1 fresh weight, respectively, and these agree with other recently published data. During the cooking process only 14–28% of the individual glucosides were retained in the cooked tissue, the remainder being largely leached into the cooking water with only a small loss attributed to the formation of the respective aglycones.

Induction of the anticarcinogenic marker enzyme, quinone reductase, in murine hepatoma cells in vitro by flavonoids

Some flavonoids induce phase II enzymes both in vivo and in vitro. We have determined the structural requirements for this activity by examining the ability of naturally-occurring flavonoids to induce the phase II enzyme, quinone reductase (NAD(P)H:quinone oxidoreductase; EC 1.6.99.2), in murine Hepalclc7 cells. Hydroxylation of the B ring is not essential for induction, since galangin and kaempferol (with 0 and 1 hydroxyl in the B ring, respectively) are better inducers than quercetin (2 B ring hydroxyls). A 2,3 double bond in the C ring is essential for induction, since taxifolin, which has the same substitution pattern as quercetin but lacks the 2,3 double bond, is not an inducer. This is supported by catechin and epicatechin, which do not possess the 2,3 double bond and are also not inducers. A 3-hydroxyl group increases the activity but is not essential for induction, since apigenin is an inducer but kaempferol (which has the same structure as apigenin but possesses a 3-hydroxyl group) is more effective. The data show that, of the flavonoids, the flavonols are the most effective inducers of quinone reductase activity in Hepa1c1c7 cells (kaempferol approximately galangin > quercetin > myricetin approximately apigenin (a flavone)) and that flavanols and flavans are ineffective.

Analytical problems in the study of flavonoid compounds in onions

Flavonoids, although potentially mutagenic, are widely thought to have beneficial effects in the diet resulting from their antioxidant and metal binding properties. Epidemiological evidence correlates diets rich in flavonoid compounds with a low risk of coronary heart disease. In the present work flavonoids (principally flavonols and anthocyanins) are studied in the onion, a major dietary source of flavonoids. This work concentrates on the development of methods to study flavonoids in their natural form in plant foods as conjugates. Two major components quercetin monoglucoside and quercetin diglucoside account for 80% of the total flavonoids in onions. Anthocyanins are only minor components of the flavonoid spectrum in the edible portion of red varieties. A preliminary study of flavonols in onions, which had been finely chopped, suggests that in most varieties there is only a small loss in total flavonol but that there is a progressive loss of the diglucoside component with an accompanying quantitative accumulation of the monoglucoside and the aglycone.

Abnormal metabolites produced by Daucus carota roots stored under conditions of stress

Abstract The formation of dihydroisocoumarins (I and II), the chromones (III and IV) and scopoletin (V) is induced in carrot roots by storage in the presence of low concentrations of ethylene and by inoculation with various fungi. There has been concern over possible toxic effects of dihydroisocoumarins and some preliminary toxicological data on (I), (II) and (IV) are included.

A comparison of the flavonol content and composition in dessert, cooking and cider-making apples; distribution within the fruit and effect of juicing

The fruit of all apple varieties tested possessed five quercetin glycosides, namely hyperin, isoquercitrin, reynoutrin, avicularin and quercitrin, as the major flavonol components. Total flavonol levels were in the range 26.4 to 73.9μg/g fresh wt (expressed as aglycone) with hyperin the dominant form in all varieties except Egremont and Jonagored, where quercitrin predominated, and the cider apples, where avicularin predominated. The proportion of flavonol in the peel ranged from 63.0 to 97.1% for the dessert and cooking apples and was not dependent on fruit size. Juice produced from the three varieties of cider apple contained 9.9 to 12.7% of the flavonols with the remainder retained in the pomace.

Antioxidant properties of flavonol glycosides from green beans

We have examined the antioxidant activity of one class of polyphenolic compounds in green beans: two novel flavonol glycosides (quercetin 3-O-[xylosyl(1-->2)]-rhamnosyl (1-->6)-glucoside and the corresponding kaempferol analogue), quercetin 3-O-glucuronide, kaempferol 3-O-glucuronide, quercetin 3-O-rutinoside and kaempferol 3-O-rutinoside. The Trolox equivalent antioxidant capacity (TEAC) and inhibition of iron/ascorbate-induced lipid peroxidation of phosphatidyl choline vesicles were measured. In the aqueous phase TEAC assay, the glucuronide and rutinoside of quercetin were good antioxidants, but not as effective as the quercetin aglycone. TEAC values for the glucuronide and other glycosides of kaempferol were much lower than the corresponding quercetin species but similar to that of the kaempferol aglycone. Quercetin 3-O-glucuronide and quercetin 3-O-rutinoside were both potent inhibitors of lipid peroxidation, in contrast to the those of kaempferol. The compounds described herein demonstrate the antioxidant activity of the flavonols present in green beans and indicate the effect on antioxidant activity of sugar substitutions in the phenolic C ring.

Formation, isolation and structure determination of methyl linolenate diperoxides

Abstract Autoxidation of methyl linolenate gives rise to isomeric mono-hydroperoxides by reaction with one mole of oxygen but further reaction with a second mole of oxygen readily occurs to produce an isomeric mixture of diperoxides. Autoxidation of individual pure methyl hydroperoxylinolenate isomers has been used as a method of obtaining less complex diperoxide mixtures which can be separated into their pure components by preparative high-pressure liquid chromatography (HPLC). The major diperoxide isomers arising from the autoxidation of pure 9R- and 13S- hydroperoxides of methyl linolenate have been isolated and characterised as isomeric epidioxyhydroperoxides of methyl linolenate. These same compounds have been identified as components of the more complex mixture of diperoxides produced during methyl linolenate autoxidation. The structures of the isolated diperoxides have been determined by physico-chemical methods and a mechanism for their formation is proposed.

Effects of saponins and glycoalkaloids on the permeability and viability of mammalian intestinal cells and on the integrity of tissue preparations in vitro.

The effects of potato and tomato glycoalkaloids and a saponin mixture from Gypsophila were investigated in cytotoxicity studies (neutral red uptake, mitochondrial MTT reduction and release of lactate dehydrogenase), using cultured cell lines of rat and human intestinal mucosal epithelium. Experiments to assess the effects of these compounds on the integrity of the intestinal epithelium were also carried out using preparations of isolated rat jejunum in vitro. By investigating the effect of these compounds on cultured cells and on intestinal tissue preparations, changes in membrane integrity, as evidenced by lactate dehydrogenase leakage in cell culture, could be confirmed in a system more relevant to the whole gut. Of the compounds tested, alpha-tomatine was consistently the most potent in all tests, and indications of a synergistic effect on membrane depolarization were observed between alpha-chaconine and alpha-solanine at total glycoalkaloid concentrations of less than 1 mM (< 0.86 mg/ml), with an optimum when the former comprised 25% of the mixture. An increase in the apparent permeability of the brush border was observed at sublethal concentrations of the compounds, and this may have important implications with respect to enhanced uptake of macromolecules, such as allergens, whose passage through the epithelium is normally somewhat restricted.

The quantitative estimation of saponin in pea (Pisum sativum L.) and soya (Glycine max)

Abstract A thin-layer chromatographic method is described for the analysis of saponins in pea and soya flours. The results are compared with those of methods using gas chromatography and high performance liquid chromatography. The levels of saponin found in the air-classified fractions of pea flour are reported. The observed levels of saponin in soya, as estimated by thin-layer and gas chromatography (0·35% and 0·33%), are much lower than that previously reported using the former approach and reasons for this discrepancy are discussed.