Genotype and harvest time influence the phytochemical quality of Fino lemon juice (Citrus limon (L.) Burm. F.) for industrial use

Two clonal selections of lemon tree (Citrus limon Burm. f. cv. Fino), named Fino-49-5 and Fino-95, were studied to ascertain the influence of genetic (clone) and environmental (season) factors on the human-health bioactive compounds of lemon juice (vitamin C and flavonoids) and the possible relationship between composition and in vitro antioxidant capacity (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis(3-ethyl-benzothiazoline-6-sulfonic acid), and ferric reducing antioxidant power) of the juice. The cultivar Fino-49-5 performed better in terms of flavonoid and vitamin C contents. Variability in the weather conditions determined, at least in part, differences in the content of lemon juice bioactives more importantly than the genetic background did. Therefore, the food industry would have phytochemically rich and nutritive lemons with practically complete independence of the harvest time and the selected cultivar.     

Online RPLC-GC via TOTAD method to isolate (+)-methyl epijasmonate from lemon (Citrus limon Burm.)

Pure (+)-methyl epijasmonate was isolated from lemon ( Citrus limon Burm.) for the first time. To that aim, two commercial essential oils and one homemade extract were included in the present paper. First, a study on the appropriate chromatographic conditions to avoid the epimerization from methyl epijasmonate to the more stable methyl jasmonate was accomplished. The results obtained are discussed. The presence of (+)-methyl epijasmonate in the three samples studied was initially established through the direct injection into GC-MS. However, the overlapping of (+)-methyl epijasmonate with other matrix components made it necessary to employ a multidimensional technique. RPLC-GC analysis via through-oven transfer adsorption-desorption (TOTAD) provided the selectivity and sensitivity required, reflecting that the homemade lemon extract was an adequate natural source to obtain pure (+)-methyl epijasmonate by means of the collection of the corresponding RPLC fraction.     

Germplasm characterization of Zolfino landraces (Phaseolus vulgaris L.) by flavonoid content

The flavonoid composition of three phenotypes of "Zolfino" landraces, a typical bean grown in Tuscany, has been elucidated by means of HPLC-DAD and HPLC-MS analysis. Flavonols, isoflavones, and anthocyanins have been separated and determined in the different samples chosen on the basis of their seed coat color. A flavonol that has not been previously found in Phaseolus vulgaris L. seeds has been characterized. The quantitative data show the presence of flavonols (ranging from 709 to 118 mg/kg of fresh weight), isoflavones (ranging from 14 to 2 mg/kg of fresh weight), and anthocyanins, in black beans only. These results show that this genotype could be very interesting from a nutritional point of view.     

Antioxidant content and free radical scavenging ability of fresh red pummelo [Citrus grandis (L.) Osbeck] juice and freeze-dried products

The antioxidative phytochemicals in various fruits and vegetables are widely recognized for their role in scavenging free radicals, which are involved in the etiology of many chronic diseases. Colored fruits are especially considered a quality trait that correlates with their nutritional values and health benefits. The specific aim of this study was to investigate the antioxidants in the juice and freeze-dried flesh and peel of red pummelo and their ability to scavenge free radicals and compare them with those in white pummelo juice. The total phenolic content of red pummelo juice extracted by methanol (8.3 mg/mL) was found to be significantly higher than that of white pummelo juice (5.6 mg/mL). The carotenoid content of red pummelo juice was also significantly higher than that in white pummelo juice. The contents of vitamin C and delta-tocopherol in red pummelo juice were 472 and 0.35 mug/mL, respectively. The ability of the antioxidants found in red pummelo juice to scavenge radicals were found by methanol extraction to approximate that of BHA and vitamin C with a rapid rate in a kinetic model. The ability of methanol extracts of freeze-dried peel and flesh from red pummelo to scavenge these radicals was 20-40% that of BHA and vitamin C effects. Fresh red pummelo juice is an excellent source of antioxidant compounds and exhibited great efficiency in scavenging different forms of free radicals including DPPH, superoxide anion, and hydrogen peroxide radicals.     

Fatty acid content and juice characteristics in black currant (Ribes nigrum L.) genotypes

The fatty acid compositions of seeds from 29 black currant genotypes were determined using a rapid small-scale procedure. There was interest in alpha-linolenic, stearidonic, and, especially, gamma-linolenic acid (GLA) contents, and most samples showed values between 11.1 and 18.7%, between 2.5 and 4.5%, and between 11.6 and 17.4%, respectively. However, six genotypes exhibited gamma-linolenic contents >18%, and values >20% were recorded in four of these genotypes. The fatty acid contents of the six genotypes were also analyzed by using a conventional procedure, and only slight differences in fatty acid composition were found between the two methods. Although GLA content was not strongly correlated with juice parameters, some genotypes had both high GLA contents and desirable juice characteristics. The results obtained provide evidence that it is possible to select for GLA contents without negatively affecting juice quality, and both aspects can be combined in a single cultivar, thereby increasing the added value of the whole fruit.     

Characterization of anthocyanins and pyranoanthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juice

Anthocyanins from blood orange [Citrus sinensis (L.) Osbeck] juices were isolated and purified by means of high-speed countercurrent chromatography and preparative high-performance liquid chromatography. Structures of the pigments were then elucidated by electrospray ionization multiple mass spectrometry and nuclear magnetic resonance (NMR) spectroscopy. The major anthocyanins of the juice were characterized as cyanidin 3-glucoside and cyanidin 3-(6"-malonylglucoside). Furthermore, six minor anthocyanins were detected and identified as cyanidin 3,5-diglucoside, delphinidin 3-glucoside, cyanidin 3-sophoroside, delphinidin 3-(6"-malonylglucoside), peonidin 3-(6"-malonylglucoside), and cyanidin 3-(6"-dioxalylglucoside). The occurrence of the latter compound in blood oranges is reported here for the first time, together with full NMR spectroscopic data. Further investigations revealed the presence of four anthocyanin-derived pigments, which are formed through a direct reaction between anthocyanins and hydroxycinnamic acids during prolonged storage of the juice. These novel pyranoanthocyanins were identified as the 4-vinylphenol, 4-vinylcatechol, 4-vinylguaiacol, and 4-vinylsyringol adducts of cyanidin 3-glucoside through comparison of their mass spectrometric and chromatographic properties with those of synthesized reference compounds.     

Ultrafiltration performance of heat-treated shamouti orange [Citrus sinensis (L.) Osbeck] juice.

During ultrafiltration of orange juice with inorganic membranes, heating of the juice prior to the filtration experiment resulted in a significant increase of the fouling indices. The effect of the irreversible fouling (Rif) was always high, whereas the reversible fouling (Rrf) depended on the treatment. It was clearly seen that fouling was reduced after pectin degradation, but the heat treatment applied to the juice before filtration still resulted in reduced fluxes. It is suggested that pectins and proteins that undergo flocculation/coagulation during the heat treatment tend to interact with the membrane-filtering layer and to cause reduction of permeation flux. To clean the membrane to restore its pure water flux, close to the initial one, a proteolitic enzyme detergent wash was needed.     

Identification of coumarins from lemon fruit (Citrus limon) as inhibitors of in vitro tumor promotion and superoxide and nitric oxide generation.

Three coumarins were isolated as siginificant inhibitors of tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced Epstein-Barr virus (EBV) activation in Raji cells from the peel of lemon fruit. They were identified as 8-geranyloxypsolaren (LE-1), 5-geranyloxypsolaren (bergamottin, LE-2), and 5-geranyloxy-7-methoxycoumarin (LE-3), respectively, by spectroscopic analysis. Three isolates had no potential O(2)-scavenging and markedly suppressed TPA-induced superoxide (O(2)(-)) generation in differentiated human promyelocytic HL-60 cells. Furthermore, LE-1 and LE-3 reduced both lypopolysaccharide (LPS) and interferon-gamma (IFN-gamma)-induced nitric oxide (NO) generation in mouse macrophage RAW 264.7 cells. Similarly, they were found to be NO generation inhibitors rather than scavengers by measuring the extracellular L-citrulline levels. The occurrence of these coumarins in a lemon fruit was abundant in the flavedo of the peel based on quantitative analysis using high-performance liquid chromatography (HPLC). The present study suggests that the coumarins in lemon fruit are promising chemopreventive agents by inhibiting radical generation.     

Involvement of proteins in cloud instability of valencia orange [Citrus sinensis (L.) Osbeck] juice.

The present study examined the involvement of proteins in cloud flocculation of Valencia orange juice. Marked differences in cloud instability were found between juices of different harvest dates. Heating of enzymatic pectin degraded juice from April and June harvests resulted in development of clumps and their precipitation. Although the juice from both harvesting dates remained hazy, the juice of April harvest was more turbid than that from June. Usually clarification increases as the temperature increases from ambient to 125 degrees C. Clarification occurred at pH 2.5-4.5 and was maximal at pH 3.5. The clarification of the April harvest juice was markedly lower than that of the June harvest. The fresh juice contained about 5.2 and 1.7 mg mL(-1) insoluble cloud matter (ICM) and alcohol-insoluble serum solids (AISS), respectively. The ICM and the AISS, respectively, contained: proteins (244.5+/-8.7 and 132+/-1.8 microg mg(-1)), galacturonic acid (40+/-0 and 120+/-0 microg mg(-1)) and neutral sugars (270+/-39 and 329+/-23 microg mg(-1)). Enzymatic pectin degradation resulted in removal of a marked portion of the pectin, and was accompanied by partial removal of neutral sugars (mainly glucose and galactose) and some proteins from the pectic polymer in both AISS and ICM. Proteins of the AISS included major bands at 10-14, 20, and 28 kDa and those of the ICM bands at 22, 24, 26, and 45 kDa.     

Involvement of proteins in cloud instability of shamouti orange [Citrus sinensis (L.) Osbeck] juice.

The present study provides evidence for the involvement of protein in cloud instability of natural orange juice. No heat-coagulable proteins were found in the serum. Insoluble cloud matter (ICM) was heat-flocculated following enzymatic pectin degradation (EPD). The degree of flocculation depended on temperature (from approximately 50 to 75 degrees C) and was highest at pH 3.5. The fresh juice contained about 6.5 and 1.8 mg mL(-1) of ICM and alcohol-insoluble solids of the serum (AISS), respectively. The ICM and the AISS contained, respectively, proteins (182+/-14 and 119+/-3 microg mg(-1)), galacturonic acid (37+/-6.6 and 175+/-1 microg mg(-1)), and neutral sugars (350+/-44 and 338+/-22 microg mg(-1)). EPD resulted in removal of a marked portion of the pectin and was accompanied by partial removal of neutral sugars (mainly glucose and galactose) and some proteins from the pectic polymer in both AISS and ICM. Under electrophoresis, proteins of the AISS included bands in the range of 20-52 kDa and 10-14 kDa and those of the ICM at 22 and 50 kDa.     

Nitrate alters the flavonoid profile and nodulation in pea (Pisum sativum L.)

A rhizosphere application of NO _inf3 ^sup- and/or naringenin affected the Pisum sativum — Rhizobium leguminosarum biovar viciae symbiosis. NO _inf3 ^sup- (5 mM) lowered while naringenin raised the nodulation status (nodule numbers and weight) and nodule efficiency (C₂H₂ reduction activity). However, the inhibitory effect of NO _inf3 ^sup- was to some extent alleviated when applied in combination with naringenin. The plant biomass was increased by the application of NO _inf3 ^sup- and naringenin, either alone or in combination, while a higher root: shoot ratio was observed only in the naringenin-treated plants. Root flavonoids are known to regulate the expression of nod genes; their high-performance liquid chromatography profile was influenced in different ways by NO _inf3 ^sup- and naringenin.     

红壤水分条件对柑橘叶片氨基酸及多胺含量的影响

本文以第四纪红黏土发育的红壤、2年生宫川温州蜜柑(Citrus unshiu Marc.cv.Miyagawa Wase)盆栽幼树为材料,采用土壤水分探头(FDR)实时监测土壤水分含量,控制土壤含水量为SWC30、SWC45、SWC60,SWC75和SWC90 5个处理(分别代表土壤最大田问持水量的30%、45%、60%、75%和90%),研究土壤水分对柑橘叶片氮含量、氨基酸含量和多胺(PAs)等氮代谢次生产物的影响.结果表明,缺水和水分过多都会降低柑橘叶片对氮素的吸收.游离氨基酸总量(y)随土壤相对含水量SWC(x)的增加而下降(y=-0.028 2x+12.049;R2=0.8524*;n=50);Pro含量(y)在SWC<75%处理的土壤水分条件下,与SWC(x)呈显著负相关(y=-0.015 2x+4.224;R2=0.860 5*;n=50):腐胺(Put)含量在SWC75处理时最低;亚精胺(Spd)含量随土壤水分增加呈抛物线变化,在SWC45处理时含量最高;精胺(Spin)含量在土壤水分SWC60处理时最高;Spd含量(y)与Put含量(x)之间呈显著正相关(y=0.240 4x2-51.337x+2 976.4;R2=0.858 6*;n=50);(Spd+Spm)/Put的比值(y)在SWC<75%的土壤水分条件时,与SWC(x)呈显著线性相关(y=0.011 2x+0.173;R2=0.851 8*;n=50).柑橘受土壤水分胁迫时产生的PAs对生理起调节作用的主要是Spd与Spin,(Spd+Spm)/Put的比值决定了柑橘受土壤水分胁迫影响的程度,初步提出该比值可能是柑橘响应土壤水分胁迫程度的一个潜在的敏感度指标.     

Effect of selenate supplementation on glycoalkaloid content of potato (Solanum tuberosum L.)

Potatoes (Solanum tuberosum L.) supplemented with increasing amounts of sodium selenate were analyzed for glycoalkaloid (GA) content. GAs were extracted with 5% acetic acid from freeze-dried tubers of two potato cultivars, Satu and Sini, harvested 10 weeks after planting as immature. The GAs alpha-solanine and alpha-chaconine were quantified by reverse-phase high-performance liquid chromatography (RP-HPLC) with diode array detection. Two independent experiments were performed. In the first experiment, the total GA concentration +/- standard error of the tubers ranged between 105 +/- 9 and 124 +/- 10 mg kg(-1) fresh weight in Satu and between 194 +/- 26 and 228 +/- 10 mg kg(-1) fresh weight in Sini. The ratio of alpha-solanine to alpha-chaconine was 0.2 in Satu and 0.5-0.6 in Sini. In the second experiment, the total GA concentration +/- standard error was 75 +/- 4 to 96 +/- 11 mg kg(-1) fresh weight, and the ratio of alpha-solanine to alpha-chaconine was 0.3-0.4 in Satu. A high sodium selenate supplementation (0.9 mg of Se kg(-1) quartz sand) slightly decreased the GA content in Satu, but this decrease was not statistically significant. Furthermore, at this addition level the Se concentration increased to a very high level of 20 microg g(-1) dry weight, which cannot be recommended for human consumption. In both experiments, the Se concentration in tubers increased with increasing sodium selenate application levels. Our results show that acceptable application levels of selenate did not have an effect on the GA concentration in immature potato tubers.     

Effect of cooking on garlic (Allium sativum L.) antiplatelet activity and thiosulfinates content

The raw form of garlic and some of its preparations are widely recognized as antiplatelet agents that may contribute to the prevention of cardiovascular disease. Herein, we examined the in-vitro antiaggregatory activity (IVAA) of human blood platelets induced by extracts of garlic samples that were previously heated (in the form of crushed versus uncrushed cloves) using different cooking methods and intensities. The concentrations of allicin and pyruvate, two predictors of antiplatelet strength, were also monitored. Oven-heating at 200 degrees C or immersing in boiling water for 3 min or less did not affect the ability of garlic to inhibit platelet aggregation (as compared to raw garlic), whereas heating for 6 min completely suppressed IVAA in uncrushed, but not in previously crushed, samples. The latter samples had reduced, yet significant, antiplatelet activity. Prolonged incubation (more than 10 min) at these temperatures completely suppressed IVAA. Microwaved garlic had no effect on platelet aggregation. However, increasing the concentration of garlic juice in the aggregation reaction had a positive IVAA dose response in crushed, but not in uncrushed, microwaved samples. The addition of raw garlic juice to microwaved uncrushed garlic restored a full complement of antiplatelet activity that was completely lost without the garlic addition. Garlic-induced IVAA was always associated with allicin and pyruvate levels. Our results suggest that (1) allicin and thiosulfinates are responsible for the IVAA response, (2) crushing garlic before moderate cooking can reduce the loss of activity, and (3) the partial loss of antithrombotic effect in crushed-cooked garlic may be compensated by increasing the amount consumed.     

Effect of shading on accumulation of flavonoid compounds in (Vitis vinifera L.) pinot noir fruit and extraction in a model system

Accumulation and compositional changes of flavonols, proanthocyanidins, and anthocyanins were measured in Vitis vinifera L. cv. Pinot noir in shaded and exposed treatments. In addition, extraction of these compounds into a model wine solution was measured. The study was conducted in a commercial vineyard within a uniform zone of relatively low vigor vines. Light exclusion boxes were installed on pairs of clusters on the same shoot (shaded treatment), and a second set of clusters on an adjacent shoot were labeled as the exposed treatment. Fruit samples were harvested at the onset of ripening (véraison) and at commercial harvest. Cluster shading resulted in a substantial decrease in mg/berry accumulation of flavonols and skin proanthocyanidins and minimal differences in anthocyanins. In analyzing seed proanthocyanidins by phloroglucinolysis, shaded and exposed treatments were similar at véraison; however, by harvest, the shaded treatment had higher extension and terminal subunits (nmol/seed) as compared to the exposed treatment. For skin proanthocyanidins, shaded fruit was lower for all subunits (nmol/berry) at both véraison and harvest. Shading caused an increase in the proportion of (-)-epicatechin and a decrease in (-)-epigallocatechin at harvest in skin extension subunits. Seed proanthocyanidins in shaded fruit contained a lower proportion of (+)-catechin and a higher proportion of (-)-epicatechin-3-O-gallate in extension subunits and a lower proportion of (+)-catechin and (-)-epicatechin-3-O-gallate and a higher proportion of (-)-epicatechin in terminal subunits. For anthocyanins, the shaded treatment had a proportional reduction in delphinidin, cyanidin, petunidin, and malvidin and a large increase in peonidin glucosides. The model extractions from the two treatments paralleled differences in the fruit with a lower concentration of flavonols, anthocyanins, and proanthocyanidins in the shaded treatment. The skin proanthocyanidin percent extraction was found to be approximately 17% higher in the exposed model extraction than the shaded treatment.     

库拉索芦荟施肥模型的建立与解析

在云南元江干热河谷气候条件下,采用二次正交旋转组合设计田间小区试验,建立了回归方程并分析了N、P、K、有机肥对库拉索芦荟鲜叶产量、芦荟甙含量和芦荟甙产量的影响。结果表明:P、K肥对提高库拉索芦荟甙含量和芦荟甙产量有显著效果,以K肥的影响最为显著,N肥和有机肥的影响不明显;N、P、K肥施用量与芦荟鲜叶产量的关系符合米氏曲线,有机肥对芦荟鲜叶产量的影响不显著。芦荟鲜叶产量最高时的施肥量为N198kghm-2、P2O5198kghm-2、K2O264kghm-2、鸡粪4125kghm-2。芦荟甙含量和芦荟甙产量最高时的施肥量为N396kghm-2、P2O5396kghm-2、K2O528kghm-2、鸡粪20625kghm-2。     

Influence of thermal treatment on black currant (Ribes nigrum L.) juice aroma

The influence of thermal treatment on black currant juice aroma was investigated in temperature and time ranges relevant for black currant juice concentration processes, namely, 45, 60, 75, and 90 degrees C. Forty-nine aroma compounds were quantified, and the thermal treatment resulted in concentration increases of most terpenes, aldehydes, furans, and phenols, whereas the concentration of esters slightly decreased. Higher temperatures and longer exposure times had larger effects on the aroma compounds. Odor triangle tests showed no sensory difference between pasteurized juice and juice heated at 60 degrees C, whereas juice heated at 90 degrees C differed significantly from pasteurized juice. It is concluded that a 90 degrees C thermal treatment of black currant juice, which is in the temperature range used for conventional evaporation of black currant juice, has an effect on the aroma and sensory properties.     

Antioxidant capacity and phenolic content in leaf extracts of tree spinach (Cnidoscolus spp.)

Total phenolic content and antioxidant capacity of two tree spinach species (Cnidoscolus chayamansa McVaugh and C. aconitifolius Miller.) were determined in raw and cooked leaf extracts. Antioxidant capacity was assessed by the oxygen radical absorbance capacity (ORAC) assay, and flavonoid glycoside composition was quantified by HPLC and identified by GC. Total phenolics and antioxidant capacity were higher in raw than in cooked leaf extracts. The ORAC values were strongly correlated with total phenolic content (r = 0.926) in all leaf extracts. The major flavonoids isolated from the leaf extracts were kaempferol-3-O-glycosides and quercetin-3-O-glycosides. C. aconitifolius leaves contained more varieties of the flavonoid glycosides than C. chayamansa. Cooking reduced antioxidant activity and phenolic content and resulted in losses of some kaempferol glycoside and quercetin glycoside residues in leaf extracts. The results of this study indicate that tree spinach leaves are a rich source of natural antioxidants for foods.     

Effects of rootstock on the composition of bergamot (Citrus bergamia Risso et Poiteau) essential oil

This paper reports the composition of bergamot oils obtained from plants grafted on the following rootstocks: sour orange, Carrizo citrange, trifoliate orange, Alemow, Volkamerian lemon, and Troyer citrange. The aim of this study is to evaluate the possibility of using rootstocks other than sour orange, checking their effect on the composition of the essential oil. Results are reported for analysis of 203 bergamot oils during the years 1997-1998, 1998-1999, and 1999-2000. The oils were analyzed by HRGC and HRGC/MS; 78 components were identified, and the results were in agreement with those reported in the literature for the Calabrian bergamot oils obtained from industry. Because of the quality of their essential oils, Alemow and Volkamerian lemon can be considered as substitutes for sour orange rootstocks.     

高接在不同基砧的锦橙上的华红脐橙的栽培表现及矿质营养含量年变化

田间调查了高接在两种不同基砧的锦橙上的华红脐橙的栽培表现及矿质营养含量年变化差异。结果表明,在栽培效应上,红桔基砧树体生长较旺,投产晚,果实小,果实含酸量较高、糖酸比较低,出汁率和Vc含量较高;枳基砧则相反。在矿质元素含量年变化上,不同基砧树的叶片矿质元素含量年变化趋势除Mn、Zn外,其它元素含量年变化趋势差异明显;不同基砧根系的各种矿质元素含量年变化趋势基本相似;在不同基砧树之间,除根系Mg、Fe含量外,叶片和根系的各种矿质元素含量在年周期内的差异明显。