Characterization and content of flavonoid glycosides in genetically modified tomato (Lycopersicon esculentum) fruits.
ABSTRACT There is a growing interest in producing food plants with increased amounts of flavonoids because of their potential health benefits. Tomatoes contain small amounts of flavonoids, most of which are located in the peel of the fruit. It has been shown that flavonoid accumulation in tomato flesh, and hence an overall increase in flavonoid levels in tomato fruit, can be achieved by means of simultaneous overexpression of the maize transcription factors LC and C1. Fruit from progeny of two modified lines (2027 and 2059) was selected for a detailed analysis and individual identification of flavonoids, at different stages of maturity. Nine major flavonoids were detected in the flesh of transgenic ripe tomatoes. LC/NMR, LC/MS, and LC/MS/MS enabled us to identify these as kaempferol-3,7-di-O-glucoside (1), kaempferol-3-O-rutinoside-7-O-glucoside (2), two dihydrokaempferol-O-hexosides (3 and 4), rutin (5), kaempferol-3-O-rutinoside (6), kaempferol-3-O-glucoside (7), naringenin-7-O-glucoside (8) and naringenin chalcone (9), which were quantified by HPLC/DAD. All but 5, 6, and 9 were detected in tomato for the first time. The total flavonoid glycoside content of ripe transgenic tomatoes of line 2059 was about 10-fold higher than that of the controls, and kaempferol glycosides accounted for 60% of this. Kaempferol glycosides comprised around 5% of the flavonoid glycoside content of ripe control tomatoes (the rest was rutin and naringenin chalcone). The rutin concentration in both transgenic and control fruits was similar.
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ABSTRACT: HPLC-DAD coupled with mass spectrometry in the positive ionization mode was applied to study the fragmentation of twelve selected flavonoids. Compounds belonging to all the major subgroups found in common plants, i.e. flavonols, flavones, dihydroflavonols, flavanones and flavanols were studied. Compound standards were injected into the spectrometer and produced characteristic mass spectra. The fragmentation of each compound was studied and it was shown that the dehydration and carbon monoxide losses from the [M+H]+ ion by the members of each subgroup produced specific fragments, thus allowing the characterization of the flavonoid subgroups. Moreover, fragments resulting from fission of the C-rings are specific of each subgroup and revealed the substitution pattern of A- and B-rings. In order to verify the identifying efficiency of the positive ionization mode through these characteristic fragmentations, the unknown flavonoids of an Origanum vulgare diethyl ether extract were separated with the HPLC system and the major peaks were successfully identified with the mass spectrometer.Molecules 02/2007; 12(3):593-606. · 2.39 Impact Factor
Article: Optimization of Total Flavonoid Compound Extraction from Gynura medica Leaf Using Response Surface Methodology and Chemical Composition Analysis.[show abstract] [hide abstract]
ABSTRACT: Optimization of total flavonoid compound (TFC) extraction from Gynura medica leaf was investigated using response surface methodology (RSM) in this paper. The conditions investigated were 30-60% (v/v) ethanol concentration (X(1)), 85-95 °C extraction temperature (X(2)) and 30-50 (v/w) liquid-to-solid ratio (X(3)). Statistical analysis of the experiments indicated that temperature and liquid-to-solid ratio significantly affected TFC extraction (p < 0.01). The Box-Behnken experiment design showed that polynomial regression models were in good agreement with the experimental results, with the coefficients of determination of 0.9325 for TFC yield. The optimal conditions for maximum TFC yield were 55% ethanol, 92 °C and 50 (v/w) liquid-to-solid ratio with a 30 min extraction time. Extracts from these conditions showed a moderate antioxidant value of 54.78 μmol quercetin/g dry material (DM), 137.3 μmol trolox/g DM for 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 108.21 μmol quercetin/g DM, 242.31 μmol trolox/g DM for 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS(+)), respectively. HPLC-DAD-MS analysis showed that kaempferol-3-O-glucoside was the principal flavonoid compound in Gynura medica leaf.International Journal of Molecular Sciences 01/2010; 11(11):4750-63. · 2.60 Impact Factor
Article: Phenolic profile and hydrophilic antioxidant capacity as chemotaxonomic markers of tomato varieties.[show abstract] [hide abstract]
ABSTRACT: Tomatoes (Solanum lycopersicum L.), the second most important vegetable crop worldwide, are a key component in the so-called "Mediterranean diet", which is strongly associated with a reduced risk of chronic degenerative diseases. In this work, we evaluate the differences in the total and individual polyphenol content and hydrophilic antioxidant capacity of seven varieties of tomato cultivated in Vegas Bajas del Guadiana, Badajoz (Spain), which were collected from two consecutive harvests (2008-2009). Hydrophilic antioxidant capacity was evaluated using the TEAC assay, while the Folin-Ciocalteau assay with a previous cleanup was used to establish total polyphenol content. The method was optimized and validated. Individual polyphenols were quantified using liquid chromatography/electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) on a triple quadrupole. All compounds were found to be significantly different when analysis of variance was performed. Results from the principal component analysis show that phenolic compounds and hydrophilic antioxidant capacity were responsible for the differences among tomato samples according to variety.Journal of Agricultural and Food Chemistry 03/2011; 59(8):3994-4001. · 2.82 Impact Factor