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In this study, the fate of Alicyclobacillus acidoterrestris spores in different types of juice concentrates stored under different conditions was investigated. In addition, the impact of dilution procedures during the enrichment step for the detection of Alicyclobacillus in lemon juice concentrates was studied. Pear, red grape, mango, tangerine, carrot and lemon juice concentrates (50–69.4°Brix, pH 1.7–4.3) were inoculated with A. acidoterrestris spores (103 spore/mL) and stored at 4 °C and 20 °C, after which the spores were counted at 0, 2, 5, 9, 17, 21, 28, 36, 43, and 50 days. No significant differences in the number of Alicyclobacillus spores were observed at storage temperatures of 4 °C and 20 °C (p N 0.05). The results also indicated that the number of spores of A. acidoterrestris remained stable in all types of juice concentrates during the storage period, except in lemon juice concentrate. In lemon juice concentrate, a decline in A. acidoterrestris spore populations of 0.3–0.8 log CFU/mL was observed within 5–10 days of storage. The decline in A. acidoterrestris spore populations was more pronounced in cloudy lemon juice concentrate, which contained higher concentrations of flavonoids (mainly eriocitrin and hesperidin) than clarified lemon juice concentrate. It was also found that dilution of lemon juice concentrate samples in the proportion of 1:19 allowed the germination of A. acidoterrestris spores and the growth of populations of up to 107 CFU/mL. In contrast, the proportion (1:9) recommended in internationally recognized methods led to a reduction in the population of this microorganism that would yield false negative results. Data presented in this study demonstrated that Alicyclobacillus spores remain stable in most juice concentrates during storage, but that natural antimicrobial compounds present in some of them may decrease spore counts and inhibit their recovery by detection procedures. Fil: Oteiza, Juan Martín. Centro de Invest. y Asistencia Tec. A la Industria; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Soto, Silvina. Centro de Invest. y Asistencia Tec. A la Industria; Argentina Fil: Alvarenga, Verônica O.. Universidade Estadual de Campinas; Brasil Fil: Sant'Ana, Anderson S.. Universidade Estadual de Campinas; Brasil Fil: Giannuzzi, Leda. Provincia de Buenos Aires. Gobernación. Comisión de Investigaciones Científicas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Criotecnología de Alimentos. Universidad Nacional de La Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Criotecnología de Alimentos; Argentina

Eriocitrin (eriodictyol 7-O-β-rutinoside) is the main flavonoid in lemon fruit. In this study, eriocitrin was investigated for its lowering effect on serum and hepatic lipids in high-fat and high-cholesterol fed rats. Rats in the control group (N= 6) were fed a 20% lard and 1% cholesterol diet for 21 d, and rats in the 0.35% eriocitrin group (N= 6) and 0.70% eriocitrin group (N= 6) were fed a diet supplemented with eriocitrin 0.35% and 0.70%, respectively. The content of hepatic total cholesterol and triglyceride in the eriocitrin group was no different from that of the control group. The total cholesterol, VLDL+LDL, triglyceride, and phospholipid in the serum of the 0.35% eriocitrin group showed significantly lower concentrations than the control group (P < 0.05), although there was no difference in the HDL concentrations among the groups. The lowering effect of eriocitrin for serum total cholesterol was thought to be caused by a decrease in VLDL+LDL. The 0.35% eriocitrin group was shown to have a significant increase in excretion of fecal bile acid (P < 0.05) and a tendency for enhanced hepatic m-RNA levels of LDL receptor in comparison with the control group.

In order to develop a database for flavanones, the dominant flavonoid class in the genus citrus, the relevant scientific literature on flavonoids in grapefruit, lemons, and limes was searched, abstracted, documented, standardized by taxons and units (mg/100 g) and examined for quality. Values for eight flavanones (didymin, eriocitrin, hesperidin, naringin, narirutin, neoeriocitrin, neohesperidin, poncirin) are presented. Grapefruit had a total flavanone content (summed means) of 27 mg/100 g as aglycones and a distinct flavanone profile, dominated by naringin. White grapefruit varieties tended to be slightly but not significantly higher in total flavanones than pink and red varieties. For lemons, total flavanones (summed means) were 26 mg/100 g and for limes 17 mg/100 g. The flavanone profiles of both lemons and limes were dominated by hesperidin and eriocitrin.

Flavanones constitute the majority of flavonoids in citrus fruits such as sweet (Citrus sinensis) and sour oranges (C. aurantium) and their near relatives—tangerines/mandarins (C. reticulata), tangors and tangelos. The relevant chemical analytic literature was searched, abstracted, documented, standardized, examined for quality, enumerated, and summarized in a database for these citrus flavanones: hesperidin, naringin, narirutin, eriocitrin, neohesperidin, didymin, neoeriocitrin, and poncirin. Sour oranges had a distinct flavanone profile dominated by naringin and neohesperidin, and were highest in total flavanones (summed means) (48 mg/100 g aglycones). Total flavanones (summed means) in sweet oranges, tangerines, and tangors were similar (∼20 mg/100 g), and hesperidin and narirutin dominated the flavanone profiles for these three fruits. Total flavanones (summed means) in tangelos (30 mg/100) were midway between sour and sweet oranges and the tangelo flavanone profile exhibited characteristics of both species. The database provides information on several varieties of citrus and eight flavanone compounds.

An inhibitory compound acting against rat platelet 12-lipoxygenase was isolated from the peel of Lumie fruit (Citrus lumia) by activity-guided separation. It was identified as eriocitrin (eriodictyol 7-O-rutinoside) by spectroscopic analyses. Eriocitrin inhibited 5-lipoxygenase (IC5029.1 µmol L−1) from rat peritoneal polymorphonuclear leukocytes in addition to 12-lipoxygenase (IC5022.3 µmol L−1). Its aglycone, eriodictyol (5,7,3′, 4′-tetrahydroxyflavanone), was a much more potent inhibitor of both 12-lipoxygenase (IC500.07 µmol L−1) and 5-lipoxygenase (IC500.20 µmol L−1). It also inhibited the production of leukotriene B4 in intact peritoneal polymorphonuclear leukocytes stimulated with calcium ionophore A23187 (IC5012.7 µmol L−1). The distribution of eriocitrin in 39 citrus fruits was investigated by high-performance liquid chromatography analysis. Lumie, eureka lemon (Citrus limon), Sambokan (Citrus sulcata), Sudachi (Citrus sudachi) and Koji (Citrus leiocarpa) fruits were found to contain high levels of eriocitrin in both peel and juice vesicles. Copyright © 2006 Society of Chemical Industry

Polyphenolic compounds such as eriocitrin, luteolin-7-O-rutinoside, diosmin, hesperidin, narirutin, isorhoifolin, rosmarinic and caffeic acids were identified in an aqueous extract (Ex) obtained from peppermint leaves (Menthae × piperitae folium). The content of polyphenols in Ex was as follows: eriocitrin 38%, luteolin-7-O-rutinoside 3.5%, hesperidin 2.9%, diosmin 0.8%, isorhoifolin 0.6%, narirutin 0.3%, rosmarinic acid 3.7% and caffeic acid 0.05%. The strongest antiradical activity (determined as DPPH• scavenging features) was observed for luteolin-7-O-rutinoside, eriocitrin and rosmarinic acid. Caffeic acid and hesperidin revealed a lower antiradical activity while isorhoifolin, narirutin and diosmin showed the lowest activity. The strongest anti-H2O2 activity was observed for eriocitrin, a little lower for rosmarinic acid. The rate of hydrogen peroxide scavenging activity displayed by luteolin-7-O-rutinoside and caffeic acid was lower than that of rosmarinic acids. Hesperidin appeared to be a very weak scavenger of hydrogen peroxide. Almost no anti-H2O2 activity was demonstrated for diosmin, narirutin and isorhoifolin. Among examined flavonoids, the strongest antiradical and anti-H2O2 activity was shown for compounds with two hydroxy groups bound to the Bring in ortho position in relation to each other. Replacement of one hydroxy group in the Bring with a methoxy group or removing one hydroxy group leads to decrease of antiradical and anti-H2O2 activity of flavonoids. Our results suggest that eriocitrin is a powerful peppermint antioxidant and a free radical scavenger.

The major flavanone-7-O-glycoside constituents in citrus fruit juices (naringin, hesperidin, neohesperidin, narirutin, and eriocitrin) were separated as diastereomers by multidimensional liquid chromatography. The method consisted of coupling two HPLC columns: a reversed-phase (RP(18)) column was used for the separation of flavanone glycosides, which were, then, individually switched into a carboxymethylated beta-cyclodextrin (beta-CD)-based column and resolved as the corresponding stereoisomers. The method was used for the full analysis of flavanone glycosides in fresh hand-squeezed and commercial fruit juices by combining the quantitative estimation with the diastereomeric analysis. Quantitative data were in general consistent with previously reported data in this field. CC-LC isomer analysis was carried out by coupling the beta-CD column with a mass spectrometer operated with negative ion electrospray ionization (ESI-MS). The results showed that hesperidin was present in orange juices almost exclusively as the 2S isomer, whereas narirutin had mainly the 2R configuration. In grapefruit juices (2S)-naringin prevailed with the respect to the 2R isomer, whereas the opposite was true for narirutin. Lemon juices contained eriocitrin stereoisomers in equal amount (50% each), but hesperidin was almost exclusively found as the 2S isomer. Significant differences of the diastereomeric ratios were observed between freshly squeezed juices and juices from commercial sources.

Some of the medicinal properties of lemons are due to the flavonoids they contain since they are involved in many biological activities and have many health-related functions. The levels of the principal flavanones and flavone found in different cultivars of Citrus limon, are analysed in an attempt to identify the most interesting as regards the content of such secondary compounds. The results show that the immature fruits from cultivars Lisbon and Fino-49 are ideal for obtaining the flavanone hesperidin, while the mature fruits of cultivar Fino-49 and the leaves of cultivar Eureka are the most interesting for obtaining the flavone diosmin and the flavanone eriocitrin.

A method was developed for the separation of diastereomers of flavanone-7-O-glycosides by capillary electrophoresis using sulfobutyl ether-β-cyclodextrin (SBE-β-CD) in the background electrolyte. The effect of the concentration of the CD additive, buffer pH, and organic modifier on the migration times and resolution for five flavanone glycosides (naringin, hesperidin, neohesperidin, narirutin, and eriocitrin) was studied. Baseline separations of these compounds as pairs of diastereoisomers were achieved with 20 mM tetraborate buffer at pH 7 containing 5 mg/mL of SBE-β-CD and 10% (v/v) of methanol. The developed method was used for the qualitative analysis of the diastereomeric composition of the major flavanone glycosides in different citrus juices. The ability of SBE-β-CD to discriminate flavanone enantiomers was also investigated.

This review discusses the relationship between the chemical composition and antioxidant property of peppermint tisane and essential oil. Phenolic acids (e.g. rosmarinic and caffeic acids), flavones (e.g. luteolin derivatives) and flavanones (e.g. eriocitrin derivatives) are possibly the major infusion antioxidants. Vitamin antioxidants (e.g. ascorbic acid and carotenoids) are minor contributors to the overall antioxidant potential. Unsaturated terpenes having a cyclohexadiene structure (e.g. terpinene) and minor cyclic oxygenated terpenes (e.g. thymol), may contribute to antioxidant potential whilst acyclic unsaturated oxygenated monoterpenes (e.g. linalool) may act as pro-oxidants in essential oil. Findings on the antioxidant potential of major cyclic oxygenated terpenes (menthol and menthone) are conflicting. Antioxidant behaviour of aqueous/organic solvent extracts and essential oil as well as the effect of environmental stresses on essential oil and phenolic composition are briefly discussed.

Purpose: To investigate 38 natural and 62 branded lemon juices for their total antioxidant activities and flavonoid content. Methods : The ferric reducing antioxidant power was applied to evaluation of antioxidant. Additionally quercetin, eriocitrin, diosmin and hesperidin contents were evaluated by high performance liquid chromatography (HPLC). Results : Total antioxidant activity of the juices ranged from 91.38 ± 0.01 to 526.93 ± 0.12 μg/ml. The maximum contents of quercetin, hesperidin and eriocitrin 14.83 ± 2.83, 104.84 ± 12.58 and 10.68 ± 2.41 μg/mL, respectively. The natural juices had diosmin levels as high as 236.18 mL. The limit of detection (LOD) ranged from 0.15 μg mg-1 for hesperidine to 0.53 μg ml-1 for diosmin while the limit of quantification (LOQ) ranged from 0.27 μg mg-1 for hesperidine to 0.93 μg ml-1 for quercetin. Conclusion: The developed method can also be applied to the determination of related extracts of orange cultivars as well as various extracts of medicinal plants. Keywords : Antioxidant, Flavonoid, Hesperidin, Diosmin, Eriocitrin, Quercetin, Lemon juice

Lemon (Citrus limon) contains various bioactive flavonoids, and prevents obesity and obesity-associated metabolic diseases. We focused on eriocitrin (eriodictyol 7-rutinoside), a powerful antioxidative flavonoid in lemon with lipid-lowering effects in a rat model of high-fat diet. To investigate the mechanism of action of eriocitrin, we conducted feeding experiments on zebrafish with diet-induced obesity. Oral administration of eriocitrin (32 mg/kg/day for 28 days) improved dyslipidaemia and decreased lipid droplets in the liver. DNA microarray analysis revealed that eriocitrin increased mRNA of mitochondrial biogenesis genes, such as mitochondria transcription factor, nuclear respiratory factor 1, cytochrome c oxidase subunit 4, and ATP synthase. In HepG2 cells, eriocitrin also induced the corresponding orthologues, and reduced lipid accumulation under conditions of lipid loading. Eriocitrin increased mitochondrial size and mtDNA content, which resulted in ATP production in HepG2 cells and zebrafish. In summary, dietary eriocitrin ameliorates diet-induced hepatic steatosis with activation of mitochondrial biogenesis.

The sonochemical effects of ultrasound (US) treatment on 14 flavonoids representing the main flavonoids in citrus fruit were investigated in a standard mixture by stability evaluation of a model system. Degradation products were further tentatively identified by Fourier transform infrared spectroscopy and high-performance liquid chromatography-ultraviolet detection-electrospray ionization tandem mass spectrometry. Thirteen flavonoids (i.e., eriocitrin, narirutin, neohesperidin, quercitrin, eridictyol, didymin, naringenin, luteolin, sinensetin, nobiletin, tangeretin, naringin, and hesperidin) were fairly stable whereas quercetin was degraded significantly by US treatment. The types of solvent and temperature used were important factors that determined the resulting degradation reactions. The degradation rate of quercetin was highest in 80% ethanol aqueous solution and decreased with increasing temperature. Longer US durations caused increases in the extent of quercetin degradation. Liquid height, ultrasonic intensity, pulse length, and duty cycle of US affected degradation rates but did not change the nature of degradation of the flavonoids. Four types of reactions occurred simultaneously for quercetin under US treatment: oxidation, addition, polymerization, and decomposition. Eight degradation products were tentatively identified as dimer, alcohol addition, oxidation, and decomposition products.

Objective: To assess the fruit of Feijoa sellowiana Berg var. coolidge and gorgiona, cultured in Sicily, for its gastroprotective effect, in association with performing phytochemical evaluation. Methods: By means of HPLC, vitamin E complex and polyphenol compounds were determined. The gastroprotective effect was investigated on ethanol-induced ulcers in rats, with sucralfate as reference drug. Samples of gastric mucosae, stained by periodic acid-Shiff and haematoxylin/ eosin, were observed by light microscopy. Results: In pulp and peel samples of both varieties of fruit α, β and γ tocopherols were identified, while δ was only in var. coolidge. In whole fruit of two varieties, catechin, eriodictyol, eriocitrin, pyrocatechol, quercetin, rutin, ellagic, gallic and syringic acid were determined. The fruit showed gastroprotective effect (ulcer index: var. coolidge=1.07, var. gorgiona=1.02). The efficacy was comparable to that of sucralfate (ulcer index 1.10). Histological examination confirmed the inhibition of ulcerogenic activity of the ethanol. Conclusions: Active principles of Feijoa sellowiana can play an important role in human diet and show beneficial effects on various diseases, particularly those caused by oxidative processes resulting in cell damage. The amount of polyphenols and vitamin E complex confirms the nutritional value of this fruit, grown in pedoclimatic condition very different from the origin area.

We could proposed that Ne-(hexanoy)lysine, HEL, become a useful biomarker for detection of oxidative stress damage occurred by exhaustive exercise. We examined the preventive effect of flavonoid compound, eriocitrin, against exercise-induced oxidative damage in rat liver. Eriocitrin administration prior to exercise significantly suppressed the increases in thiobarbituric acid-reactive substance caused by lipid peroxidation during exhaustive exercise. The increase in the contents of HEL in rat liver was also abolished by eriocitrin administration. The concentration of oxidized glutathione was significantly increased by exercise, but the eriocitrin administration suppressed this increase. These results suggested that eriocitrin administration prior to exercise prevented oxidative damages caused by exhaustive exercise-induced oxidative stress. Therefore, it was suggested that HEL could be a good biomarker for oxidative stress, especially at earlier stage when oxidative damage was occurred by lipid peroxidation than a stage of harmful aldehyde formation. Moreover, it was suggested that eriocitrin metabolites, eriodictyol and 3, 4 – dihydroxyhydrocinnamic, might scavenge free radicals and reactive oxygen species, resulting in suppression of lipid peroxidation and reactive proteins with radicals to form HEL. These findings implied that eriocitrin might be useful as an anti-oxidative compound to protect oxidative stress damages.

The monoclonal antibody to N(epsilon)-(hexanonyl)lysine (HEL), a novel adduct formed by the reaction of linoleic acid hydroperoxide and lysine, has been prepared and characterized. The obtained antibody specifically recognized the HEL moiety. Using the monoclonal antibody, we evaluated the protective effects of feeding eriocitrin, which is one of flavonoids in lemon fruit, on oxidative modification induced by exercise in rats. The supplementation of eriocitrin significantly suppressed the increase in HEL in the skeletal muscle by exercise. The result suggests that the determination of HEL may be a good method for evaluation of the protective effect of beneficial food factors against oxidative stress.

The effects of lemon flavonoids, as crude flavonoids prepared from lemon juice, were investigated in diabetic rats. The oxidative stress of eriocitrin (eriodictyol 7-O-beta-rutinoside) and hesperidin (hesperetin 7-O-beta-rutinoside) on streptozotocin-induced diabetic rats was investigated. Diabetic rats were given a diet which contained 0.2% crude flavonoids, 0.2% eriocitrin, and 0.2% hesperidin. After the 28-d feeding period, the concentration of the thiobarbituric acid-reactive substance in the serum, liver, and kidney of diabetic rats administered crude flavonoids, eriocitrin, and hesperidin significantly decreased as compared with that of the diabetic group. The levels of 8-hydroxydeoxyguanosine, which is exchanged from deoxyguanosine owing to oxidative stress, in the urine of diabetic rats administered eriocitrin and hesperidin significantly decreased as compared with that of the diabetic rat group. Crude flavonoids, eriocitrin, and hesperidin suppressed the oxidative stress in the diabetic rats. These results demonstrated that dietary lemon flavonoids of eriocitrin and hesperidin play a role as antioxidant in vivo.

The flavonoids present in the wastes and residues obtained during the industrial processing of lemons were characterized and quantified. Selected products of the juice-extraction line and essential-oil-recovery line were studied. Industrial lemon juice is characterized by containing hesperidin and eriocitrin as the main flavonoid glycosides (130 μg/ml), other minor flavanones and C-glycosylflavones. After the concentration process, the concentrated juice contained a 5-fold increase in the concentration of flavonoids, but some of the hesperidin present in the initial juice was lost. The flavanone glycosides were mainly concentrated in the peel and solid residues, where hesperidin was present as the prevailing flavonoid with concentrations of up to three times higher than those of eriocitrin. In liquid residues eriocitrin was the main flavonoid. The solid residues (peel and frit precipitates) were the most interesting sources for the industrial recovery of flavonoids (13 kg/t processed lemons).

We investigated the antioxidative flavonoid glycosides in the peel extract of lemon fruit (Citrus limon). Six flavanon glycosides: eriocitrin, neoeriocitrin, narirutin, naringin, hesperidin, and neohesperidin, and three flavone glycosides: diosmin, 6,8-di-C-β-glucosyldiosmin (DGD), and 6-C-β-glucosyldiosmin (GD) were identified by high-performance liquid chromatography (HPLC) analysis. Their antioxidative activity was examined using a linoleic acid autoxidation system. The antioxidative activity of eriocitrin, neoeriocitrin and DGD was stronger than that of the others. Flavonoid glycosides were present primarily in the peel of lemon fruit. There was only a small difference in the content of the flavonoid glycosides of the lemon fruit juice from various sources and varieties. Lemon fruit contained abundant amounts of eriocitrin and hesperidin and also contained narirutin, diosmin, and DGD, but GD, neoeriocitrin, naringin, and neohesperidin were present only in trace amounts. The content of DGD, GD, and eriocitrin was especially abundant in lemons and limes; however, they were scarcely found in other citrus fruits. The content of flavonoid compounds in lemon juice obtained by an in-line extractor at a juice factory was more abundant than that obtained by hand-squeezing. These compounds were found to be stable even under heat treatment conditions (121°C, 15 min) in acidic solution.

The effects of lemon juice and its crude flavonoids on blood pressure were examined using spontaneously hypertensive rats (SHR). The 5% diluted lemon juice was orally administered in the diet to SHR, and they tended to have a lower systolic blood pressure than the control rats after 90 days. The systolic blood pressure of SHR fed a diet containing crude flavonoids from the juice for 16 weeks was significantly lower than that of the control group (p

An antioxidant was isolated from the peel and juice of lemon fruit (Citrus limon BURM. f.). It was identified as eriocitrin (eriodictyol 7-rutinoside) of the flavanone glycoside by HPLC, 1H-NMR and 13C-NMR analyses. The purified eriocitrin was readily soluble in water, methanol, and ethanol. A water solution of 0.05% eriocitrin was weakly acidic (pH 4.2). Eriocitrin was found to be stable even at high temperature (121°C, 15 min) in acidic solution (pH 3.5). The distribution of eriocitrin in citrus fruits was found to be especially abundant in lemons and limes, however, it was scarcely found in other citrus fruits. In the case of lemon fruit, eriocitrin was primarily distributed in the peel (about 1,500 ppm) composed of the albedo (mesocarp), flavedo (epicarp), and pulp vesicles. It was also significantly present in the juice (about 200 ppm) but was not detected in the seed. Two varieties of lemon fruits, eureka and lisbon, almost had the same eriocitrin content. The antioxidative activity of eriocitrin in the linoleic acid autoxidation system was equal to that of α-tocopherol, and it was enhanced when used together with citric acid. The eriocitrin had a synergistic effect on α-tocopherol.

Reversed-phase-HPLC analysis by means of superficially porous silica particle columns (fused-core) was applied to the investigation of flavonoids, coumarins, and psoralens in lime juice samples. Hesperidin (367.0 ± 16.0 ppm) and eriocitrin (148.0 ± 7.9 ppm) were the most abundant flavonoids. Fifteen coumarins and furocoumarins were determined, including bergamottin (29.6 ± 1.1 ppm), 5-geranyloxy-7-methoxycoumarin (16.5 ± 0.6 ppm), and oxypeucedanin hydrate (9.9 ± 0.5 ppm) as predominant compounds. These molecules are today well known for their beneficial effects on human health. As a consequence, the present study, beyond investigating for the first time the chemical composition of lime juice, highlights also its health-promoting qualities, due to its content of flavonoids and coumarins.

Objective To evaluate the chemopreventive potential of phenolic compounds – potassium apigenin, cocoa, catechins, eriocitrin and rosmarinic acid in oral carcinogenesis induced in hamsters by means of the topical application of 7,12-dimethylbenz(a)anthracene(DMBA). Study design An experimental study at the University of Murcia. Methods 50 male Syrian hamsters (Mesocricetus auratus) were divided into five groups of ten: Group I (control group): 0.5% DMBA; Group II: 0.5% DMBA + 1.1 mg/15 ml potassium apigenin; Group III: 05% DMBA + 2.5 mg/15 ml cocoa catechins; Group IV: 0.5% DMBA + 6 mg/15 ml eriocitrin; Group V: 0.5% DMBA + 1.3 mg/15 ml rosmarinic acid. The flavonoids were administered orally. All the animals were sacrificed after 12 weeks. Macroscopic, microscopic and immunohistochemical (PCNA and p53) analyses of the lesions were performed. Results All the groups treated with phenolic compounds showed lower incidences of tumour, greater differentiation and lower scores in the tumour invasion front grading system in comparison with the control group. Potassium apigenin and rosmarinic acid achieved the best results, the former considerably reduced the carcinoma tumour volumes developed and both significantly reduced the intensity and aggression of the tumours. Immunoexpression of PCNA and p53 were significantly altered during DMBA-induced oral carcinogenesis. Conclusions Animals treated with phenolic compounds, particularly potassium apigenin and rosmarinic acid, showed a lower incidence of tumours.

In the present study, for the first time, a simplified miniaturized ultrasound-assisted matrix solid-phase dispersion (SM-USA-MSPD) method with a different application for liquid matrices was developed to extract different flavonoids (hesperidin, diosmin, eriocitrin, narirutin, naringin, hesperetin and naringenin) from citrus fruit juice and human fluid samples prior to their determination using high performance liquid chromatography (HPLC). Different effective parameters were studied and under the optimum conditions (including sample volume: 150μL; solid phase: silica-based C18, 200mg; eluting solvent: methanol, 500μL; pH: 4; and sonication: 6min; at room temperature), limits of detection and limits of quantification were ranged from 23.3 to 46.8ngmL(-1) and 74.8 to 141.5ngmL(-1), respectively. Once optimized, analytical performance of the method was studied in terms of linearity (0.074-198.5μgmL(-1), r(2)>0.991), accuracy (recovery=84.6-101.5%), and precision (repeatability: intra-day precision

High-performance liquid chromatography (HPLC) coupled with ultraviolet-visible spectrophotometry using a photodiode-array detector was used as a routine method for the simultaneous separation and determination of 25 naturally occurring Citrus flavonoids. The separation system consisted of a C18 reversed-phase column, a gradient system of 0.01 M phosphoric acid (A) and methanol (B), and a photodiode-array detector. Each of the 25 flavonoids was eluted from the column with a gradient system composed of three periods: (1) 0–55 min, 70-55% (v/v) A in B, (2) 55–95 min, 55-0% A in B, and (3) 95–100 min, isocratic, 100% B, and quantified by spectrophotometric detection at 285 nm. Identifications of specific flavonoids were made by comparing their retention times (tR) and UV spectra with those of standards. The relative standard deviations of tR, values were 0.029–0.321%. The recoveries of pure eriocitrin, naringin, hesperidin and tangeretin added to tissues prepared from Unshiu (Citrus unshiu Marc.) and Hirado-buntan (Citrus grandis Osbeck f. Hirado) and subsequent extraction were 97.47–103.13% from the mesocarp and 96.87–104.93% from the juice with standard deviations of 2.32–5.72% and 2.18–5.96%, respectively.

A reversed-phase high-performance liquid chromatographic method has been developed for the qualitative and quantitative analysis of a caffeic acid derivative, rosmarinic acid, and the main flavone and flavanone glycosides in leaves of Mental×piperita, namely eriocitrin, luteolin 7-O-rutinoside, hesperidin, isorhoifolin, diosmin, eriodictyol 7-O-glucoside, and narirutin. The last two were identified for the first time in M.×piperita and Mentha genus, respectively. Forty cloner, belonging to the three varieties of M.×piperita, were investigated. The eight phenolic constituents are present in all clones. The flavonoid content of the leaves ranged from 8.6 to 11.8% on dry weight basis

The flavonoid composition of crude Citrus limetta Risso (limetta, Mediterranean sweet lemon) juice was elucidated by means of reverse-phase HPLC–DAD–ESI-MS/MS analysis. In a single course, eight compounds (C- and O-glycosyl flavonoids) were identified, six of which were found for the first time in limetta juice: four C-glucosyl flavones, namely vicenin-2, lucenin-2 4′-methyl ether, orientin 4′-methyl ether and scoparin; the O-glycosyl flavone, rhoifolin; and the O-glycosyl flavanone, eriocitrin. In addition, ABTS + and DPPH radical quenching and FRAP assays demonstrate that limetta juice possesses remarkable antioxidant activity.

The effect of high-temperature-conditioning treatments (1-2 days at 37°C) on fruit quality, flavonoids, total antioxidant capacity (TAC) and vitamin C was investigated in chilling-sensitive 'Fortune' mandarins subjected to single or double quarantine treatments (16 or 32 days at 1.5°C, respectively). High temperature-conditioning treatments, which reduced chilling injury, allowed fruits to withstand quarantine treatments without affecting the fruit quality, vitamin C or TAC. Hesperidin and isorhoifolin were the most abundant flavonoids followed by didymin and narirutin, whereas the polymethoxylated flavones (PMFs) nobiletin and tangeretin were the less abundants. Didymin and narirutin slightly increased (~1.5-fold) at 1.5°C. A 4-fold increase occurred in eriocitrin, though its concentration was much lower. Small differences in flavonoids were found between non-conditioned fruit and fruit conditioned for 1 day after cold storage and their concentration in carpellary membranes were, in general, much higher than in juice. Therefore, fruit conditioning at 37°C allows chilling-sensitive citrus cultivars to withstand quarantine treatments without having deleterious effects on the fruit quality, vitamin C or relevant flavonoids., This work was supported by research Grants FAIR-CT-98-4096 from the EU and AGL2002-1227, AGL2006-09496 and CONSOLIDER 2007-00063 from the Comisión Interministerial de Ciencia y Tecnología (CICYT, Ministerio de Ciencia y Tecnología), Spain, and PROMETEO2010/010 from the Generalitat Valenciana.

Phenolic glucosides having radical scavenging activity were examined from the fraction eluted with 20% methanol on Amberlite XAD-2 resin applied to lemon (Citrus limon) juice by using reversed phase chromatography. Four phenolic glucosides were identified as 1-feruloyl-beta-D-glucopyranoside, 1-sinapoyl-beta-D-glucopyranoside, 6,8-di-C-glucosylapigenin and 6,8-di-C-glucosyldiosmetin by (1)H-NMR, (13)C-NMR, and MS analyses. They exhibited radical scavenging activity for 1,1-diphenyl-2-picrylhydrazyl (DPPH) and superoxide, although the activity was low in comparison with eriocitrin, a potent antioxidant in lemon fruit, and the eriodictyol of its aglycone. The phenolic compounds in lemon juice were examined for their suppressive effect on the expression of blood adhesion molecules by measuring the expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs) induced by necrosis factor-alpha (TNF-alpha). 6,8-Di-C-glucosylapigenin, apigenin, and diosmentin of the flavones were found to significantly suppress the expression of ICAM-1 at 10 muM (P0.05). The phenolic glucosides isolated in this study were contained in comparative abundance in daidai (Citrus aurantium) and niihime (Citrus unshiu x Citrus tachibana) among the sour citrus juices.

[SPA] Los hongos del género Penicillium son responsables de importantes pérdidas durante los procesos de post-cosecha de los cítricos. Así mismo se ha descrito que diversos compuestos fenólicos presentes en los frutos cítricos pueden estar implicados en los mecanismos de defensa natural desarrollado por el género Citrus frente a hongos patógenos. En este trabajo se analiza el efecto de algunos tratamientos habituales durante la post-cosecha de los frutos cítricos como son el tratamiento con ácido giberélico, la conservación en cámara fría y el tratamiento en tambor, sobre los niveles de flavonoides y resistencia a Penicillium digitatum en frutos de limón Fino-49. Los resultados obtenidos ponen de manifiesto que el tratamiento con 6 ppm de ácido giberélico, la conservación en cámara fría y los tratamientos con tambor producen incrementos en los niveles de algunos compuestos flavónicos, respecto a los niveles observados en los correspondientes controles. Entre los compuestos fenólicos presentes en los frutos de limón cabe destacar las flavanonas hesperidina y eriocitrina así como la flavona diosmina. Los estudios in vivo e in vitro llevados a cabo revelan que tanto las flavanonas como la flavona reducen el crecimiento micelial de Penicillium digitatum, produciendo alteraciones morfológicas y ultraestructurales de sus hifas. El papel de dichos compuestos fenólicos como posibles fitoalexinas, en el mecanismo de defensa de esta especie cítrica frente a Penicillium digitatum, es discutido. [ENG] Fungi of the genera Penicillium are responsible for substantial losses in citrus fruit during post harvest processes. Several phenolic compounds have been implicated in the natural defence mechanisms developed by Citrus against pathogenic fungi. In this study, we analyse the effect of some common post harvest treatments in citrus, such as 6 ppm gibberellic acid treatments, cold storage and mechanical test, on flavonoid levels and resistance to Penicillium digitatum in fruits of Fino-49 lemon. These treatments were seen to increase the levels of flavonoids compared with their respective controls. Among the phenolic compounds present in lemon, the flavanones, hesperidin and eriocitrin, and the flavone, diosmin, are of note. “In vivo” and “in vitro” studies showed that both the flavanones and flavone reduced the mycelial growth of Penicillium digitatum, in which morphological and ultrastructural alterations of the hyphae were noted. The possible role of these phenolic compounds as phytoalexins in the defence mechanism of lemon against Penicillium digitatum is discussed. Proyecto AGR/8025/06 Consejería de Agricultura, Agua y Medio Ambiente de la Región de Murcia

application/pdf, 学術雑誌論文, The concentrations of metabolites in human plasma after ingestion of flavanone glycosides (FG) and their aglycones (FA) in lemon were examined. FG consisting abundantly of eriocitrin were prepared from lemon peel and FA consisting abundantly of eriodictyol were prepared from FG by treatment with β-glucosidase. Eriodictyol, homoeriodictyol, and hesperetin in plasma up to 4 h after ingestion of FG with water or FA with water by subjects were not detected in plasma of non-enzyme treatment but in plasma after treatment with β-glucronidase and sulfatase. Metabolites in plasma after ingestion of FG and FA in humans were shown to exist as the glucuro- and/or sulfo-conjugates of eriodictyol, homoeriodictyol, and hesperetin. After ingestion of FA, the concentration of metabolites in plasma exhibited a high maximum peak at 1 h. The AUC (area under the blood concentration time curve) level of metabolites of FA was higher than that of FG. FA were suggested to be absorbed faster and in higher amounts than FG. The AUC of metabolites in subject plasma after ingestion of FG with flavonoid-depleted lemon juice was shown to change\ to a low level in comparison with that of FG with water. The maximum concentration peak of metabolites in plasma was faster at 0.5 h than FA with water but the AUC level was similar to FA with water, when subjects ingested FA with vodka (40% ethanol). The absorption hour of FG and FA was shown to be affected by the co-existing solution.

Cyclopia subternata is one of the 24 Cyclopia species that are used to brew honeybush tea, a unique South African herbal beverage with a pleasant taste and flavor. It contains various antioxidants, very low tannin content, and no caffeine. Many health properties are associated with regular consumption of the tea. Honeybush infusions have been noted as a tonic for colds and influenza, catarrh, and pulmonic tuberculosis and is becoming well-known for its effectiveness in alleviating menopausal symptoms in women. "Unfermented" leaves of C. subternata contain pinitol, shikimic acid, p-coumaric acid, 4-glucosyltyrosol, epigallocatechin gallate, the isoflavone orobol, the flavanones hesperedin, narirutin, and eriocitrin, a glycosylated flavan, the flavones luteolin, 5-deoxyluteolin, and scolymoside, the xanthone mangiferin, and the flavonol C-6-glucosylkaempferol. The structures were elucidated by spectroscopic and spectrometric analysis.

The distribution of seven flavonoids, eriocitrin, luteolin 3'-O-beta-d-glucuronide, hesperidin, diosmin, isoscutellarein 7-O-glucoside, hispidulin 7-O-glucoside, and genkwanin, has been studied in Rosmarinus officinalis leaves, flowers, stems, and roots during plant growth. The maximum level reached by luteolin 3'-O-beta-d-glucuronide in leaves during June-August suggests the existence of a delay between the activation of the enzymes involved in the flavanone and flavone biosynthesis. The presence of hesperidin and diosmin in the vascular system is significant, and hesperidin shows even higher levels than the phenolic diterpenes and rosmarinic acid. The distribution of flavonoids observed in R. officinalis suggests a functional and structural relationship between phytoregulators and flavonoids, where flavonoids would be "protectors" of the activity of phytoregulators. A hypothesis for the general pathway of biosynthesis of these compounds in plants of the family Labiatae is proposed.

WOS: 000223137000013, PubMed ID: 15291467, An on-line high-performance liquid chromatography-1,1-diphenyl-2-picrylhydrazyl (HPLC-DPPH.,) method has been improved for the detection of polar and nonpolar radical scavenging compounds in complex plant extracts. Nine water extracts were prepared from different Mentha species, varieties, hybrids, and cultivars. After the components within each extract had been separated by reverse phase chromatography using 10-100% methanol with 2% acetic acid as a mobile phase, analytes within the eluent capable of scavenging a citric acid-sodium citrate-buffered methanol 1,1-diphenyl-2-picrylhydrazyl solution were detected by postcolumn derivatization at 517 nm. The HPLC-DPPH. on-line method was applied to the qualitative and quantitative analysis of Mentha extracts. There was a strong correlation between the scavenging (negative) peak area and the concentration of the radical scavenging reference substances used. The minimum detectable concentration (mug/mL) of the antioxidant compounds was determined. Caffeic acid, eriocitrin (eriodictyol-7-O-rutinoside), luteolin7-O-glucoside, and rosmarinic acid were identified as the dominant radical scavengers in these extracts by this method.

8 pages, 5 figures, 2 tables.-- Printed version published 2004., The grafting of the rootstock with the lemon tree is an agronomical technique used to improve production and/or quality of the fruit. The interstock has been used with different fruit trees to modulate the tree size, fruit production and quality, and the aging of the tree. The lemon trees grafted with interstocks increase their longevity, lemon production and quality; interstocks are also used to decrease the thickness of the trunk at the grafting point. This enlarging of the trunk provokes a decrease of the sap flow. In our study, “Verna” lemon trees were grafted with interstock between the rootstock and the lemon tree to follow the flavonoid content of the lemon juice. The lemon juice was obtained from the lemons collected of the grafted lemon trees. Two types of rootstocks were used: Citrus aurantium L. and Citrus macrophylla L. Seven interstocks from five cultivars of orange tree, one cultivar of lime tree, and one cultivar of tangerine tree were used. “Verna” lemon trees were also grafted directly to the rootstock. The rootstock was more important agronomic factor than the interstock on the total flavonoid content of lemon juice. The interstock grafting had only a small influence on the flavonoid content of the lemon juice, and it modulated the individual flavonoid content. Citrus aurantium L. rootstock and “Berna” and “Washington Navel” interstocks were the most appropriate to graft in the lemon tree. This interstock grafting technique does not increase the flavonoid content of the lemon juice. Regarding the individual flavonoids, the 6,8-di-C-glucosyl diosmetin was the most affected flavonoid by the type of rootstock used. The interstock used is able to alter the individual quantitative flavonoid order of eriocitrin, diosmin, and hesperidin. In addition, the HPLC-ESI/MSn analyses provided the identification of two new flavonoids in the lemon juice: Quercetin 3-O-rutinoside-7-O-glucoside and chrysoeriol 6,8-di-C-glucoside (stellarin-2). The occurrence of apigenin 6,8-di-C-glucoside (vicenin-2), eriodictyol 7-O-rutinoside, 6,8-di-C-glucosyl diosmetin, hesperetin 7-O-rutinoside, homoeriodictyol 7-O-rutinoside and diosmetin 7-O-rutinoside was also confirmed in lemon juice by this technique., This work has been funded by the Consejería de Agricultura de la Región de Murcia and the UE (FEDER) (AGR/12/FS/02). A.G.I. is holder of a research grant from the Fundación Cajamurcia.

High-performance liquid chromatography-diode array detector (HPLC-DAD) coupled with electronspray mass spectrometry (ESI-MS) was used to detect the flavonoid profile in lemon juices obtained from the main Sicilian cultivars (Femminello comune, Monachello, and Interdonato). Significant amounts of an unusual constituent were found in the lemon juice of the above-mentioned cultivars together with eriocitrin, hesperidin, and diosmin. Following purification by preparative HPLC, the structure was assigned as 6,8-di-C-glucopyranosyldiosmetin by means of DAD-UV, ESI-MS-MS, and (1)H NMR analysis. Three other minor components were also detected. One of these presented a flavone nature, and spectral data and literature references both suggested a 6,8-di-C-glucopyranosylapigenin structure. The different contents of eriocitrin, hesperidin, diosmin, and above minor components in the cultivars allow juices to be readily differentiated.

Six flavonoid glycosides, eriocitrin (1), narirutin (2), hesperidin (3), luteolin-7-O-rutinoside (4), isorhoifolin (5), diosmin (6), rosmarinic acid (7) and 5,7-dihydroxycromone-7-O-rutinoside (8), were isolated from the aerial part of Mentha piperita L. Among these compounds, compound 4 showed a potent inhibitory effect on histamine release induced by compound 48/80 and antigen-antibody reaction. This compound was more effective than luteolin and luteolin-7-O-glucoside in inhibiting histamine release from rat peritoneal mast cells. Compound 4 also caused a dose-related inhibition of the antigen-induced nasal response and significant effects were observed at doses of 100 and 300 mg/kg. These results indicate that compound 4 may be clinically useful in alleviating the nasal symptoms of allergic rhinitis.

The diastereomers of six major flavanone-7-O-glycosides (naringin, prunin, narirutin, hesperidin, neohesperidin and eriocitrin) were completely separated for the first time by chiral capillary electrophoresis (CE) employing various buffers and chiral selectors on the basis of the results achieved in 1998 in our research group by Mellenthin. The following chiral additives to the background electrolyte (BGE) were examined: native cyclodextrins (CDs; alpha-, beta- and gamma-CD), neutral cyclodextrin derivatives (dimethyl-beta-CD, hydroxypropyl-beta-CD, hydroxypropyl-gamma-CD) and charged cyclodextrin derivatives (carboxymethyl-beta-CD, carboxyethyl-beta-CD). The effect of CD type, CD concentration and pH value on chiral recognition will be discussed in the following article. In this work, lemon juice (Citrus limon L.) was also examined by chiral CE. Eriocitrin and hesperidin could be identified as characteristic flavanones and chiral separation of their diastereomers could be achieved according to the developed chiral method by capillary electrophoresis using a 0.2 M borate buffer at pH 10.0 and with 5 mM gamma-CD as chiral selector.

Eriocitrin, a flavonoid glycoside present in lemon fruit, is metabolized in vivo to a series of eriodictyol, methylated eriodictyol, 3,4-dihydroxyhydrocinnamic acid, and their conjugates. Plasma antioxidant activity increased following oral administration of aqueous eriocitrin solutions to rats. Eriocitrin metabolites were found in plasma and renal excreted urine through HPLC and LC-MS analyses. Eriocitrin was not detected in plasma and urine, but eriodictyol, homoeriodictyol, and hesperetin in their conjugated forms were detected in plasma of 4.0 h following administration of eriocitrin. In urine for 24 h, both nonconjugates and conjugates of these metabolites were detected. 3,4-Dihydroxyhydrocinnamic acid, which is metabolized from eriodictyol by intestinal bacteria, was detected in slight amounts with each form in 4.0-h plasma and 24-h urine. Eriocitrin was suggested to be metabolized by intestinal bacteria, and then eriodictyol and 3,4-dihydroxyhydrocinnamic of its metabolite were absorbed. Following administration of eriocitrin, plasma exhibited an elevated resistance effect to lipid peroxidation. Eriocitrin metabolites functioning as antioxidant agents are discussed.

Flavonoids are common in fruits and vegetables and many function as natural antioxidants, free radical scavengers, and may also chelate pro-oxidant metals, reducing their capacity to produce free radicals (Affany et al., 1987). It has been estimated that humans consume 1 gram per day of flavonoids (Kuhnau et al., 1976). The majority of citrus flavonoids are flavanones bound as glycosides (Figure 1). Hesperidin, a glycoside of hesperetin, is predominant in orange and tangerine, whereas naringin and narirutin, glycosides of naringenin, are most abundant in grapefruit. Eriocitrin and hesperidin are the predominate flavonoids found in lemon and lime. Some therapeutic effects have been reported for citrus flavanoids (Fisher et al., 1982). In 1936 citrus flavonoids were reported to decrease capillary fragility and improve blood flow and were actually labeled “Vitamin P” (Rusznyak and Szent-Gyorgyi, 1936). But further research failed to substantiate that abnormal capillary permeability was due to a deficiency in bioflavonoids. In 1950, the Federation of American Societies for Experimental Biology recommended that the term “Vitamin P”, which had been applied to flavanoids, be discontinued (Vickery et al., 1950).

South African herbal teas, rooibos and honeybush, are increasingly enjoyed as healthy alternatives to Camellia sinensis teas. They contribute to the diet with bioactive phytochemicals not commonly found in foods. Major compounds of rooibos are the unique dihydrochalcone, aspalathin, and its flavone isomers, orientin and isoorientin. Honeybush contributes the xanthones, mangiferin and isomangiferin and the flavanones, eriocitrin, narirutin and hesperidin. All these compounds are either C-glucosides or O-rhamnoglucosides, which are poorly absorbed. Phase II metabolism and degradation by intestinal bacteria are important factors in their absorption. Modulation of drug metabolising enzymes is indicated which not only could affect the therapeutic window of drugs, but also the bioavailability of other dietary flavonoids.

Seven extracts were prepared from Mentha x piperita (peppermint) leaves in sequence using a Soxhlet apparatus, viz. (40-60°) light petroleum (PE), dichloromethane (CH2Cl2), acetonitrile (ACN), ethyl acetate (EtOAc), methanol (MeOH), n-butanol and water (H2O) extracts. The phenolic and flavonoid content of each extract were estimated using spectrophotometric methods whilst a qualitative-quantitative analysis was made by reverse-phase high performance liquid chromatography coupled with photodiode array detection (HPLC-PDA). Each extract was assessed in a battery of six antioxidant-related assays so as to determine their iron(III) reductive, iron(II) chelating and free radical scavenging abilities. The MeOH-soluble extract contained the greatest content of total phenols and flavonoids based upon the Folin-Ciocalteu and 2-aminoethyl diphenylborate reagent data and HPLC-PDA analysis. Based upon the chromatographic and UV-spectral data, the leaves principally contained the cinnamic acid caffeic acid, the depside rosmarinic acid and flavonoids (flavones and flavanones). Eriocitrin (383.3 ± 2.2 mg/g extract) and rosmarinic acid (381.2 ± 1.9 mg/g extract) were the most abundant components identified within the leaves, whilst naringenin-7- O-glucoside (0.8 ± 0.01 mg/g extract) was the least abundant component identified being found only in the EtOAc-soluble extract. The EtOAc, ACN and H2O-soluble extracts demonstrated the most potent iron(III) reductive and 1,1′-diphenyl-2-picrylhydrayl, 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonate) and hydroxyl free radical scavenging properties; however, the H2O and CH2Cl2-soluble extracts were the most potent extracts in the β-carotene-linoleic acid bleaching inhibition assay. In terms of iron(II) chelation – an important antioxidant property - the PE, MeOH and H2O extracts demonstrated moderate iron(II) chelating activity.

The occurrence and distribution of flavanone glycosides in the leaves and fruits of many kinds of artificial citrus hybrid plants were investigated by polyamide thin-layer chromatography. The citrus hybrids can be divided into two broad categories, a) those containing rutinosyl glycosides, b) those containing neohesperidosyl glycosides in accordance with the case of natural citrus species. The fiavonoid patterns of rutinosyl glycosides are classified into the following groups, a) hesperidin, b) narirutin, c) hesperidin and narirutin, d) didymin and narirutin, e) hesperidin, narirutin and eriocitrin and f) hesperidin and eriocitrin, while the pattern of neohesperidosyl glycosides fall into six groups, a) naringin, b) neohesperidin and naringin, c) neohesperidin, naringin and neoeriocitrin, d) neohesperidin and neoeriocitrin, e) naringin and neoeriocitrin, and f) poncirin, neohesperidin, naringin and neoeriocitrin. It is worthy of note that a hybrid (accession number 1088) between C. unshiu and C. hassaku c...

Concentrations and HPLC profiles of the major flavonoids of lemon juice vesicles ( Citrus limon cv. Eureka) were determined for tree-grown fruit 2–55 mm diameter and in 25–30 mm diameter fruit halves cultured in vitro for up to four months. In tree-grown fruit, the total amount of hesperidin per lemon accumulates rapidly from fruit set to its maximum value at the 25–30 mm diameter stage; and thereafter the concentration continually decreases as the fruit increases in diameter. At the 25–30 mm stage, the total amount of eriocitrin begins to increase rapidly and continues until the fruit reaches full size. The amount of diosmin per fruit increases gradually throughout the development of the lemon. Flavonoids of juice vesicles from in vitro cultured fruit halves show a pattern of development similar to fruit grown on the tree. In contrast, flavonoid patterns from callus derived from juice vesicles were quite dissimilar to those of developing fruit.

A procedure has been devised to quantitate the flavanone, eriodictyol, and its glycoside, eriocitrin, by use of the color reagent, 2-aminoethyl diphenylborate. Also, some parameters in the use of this reagent in quantitative analysis have been studied. Application of the procedure to lemon bioflavonoid complex gave an average content of 2.5% eriocitrin.