裙带菜中褐藻糖胶的提取纯化及其组成的研究

该实验优化了裙带菜褐藻糖胶的提取工艺,对粗多糖进行了分离纯化,并研究了各组分的单糖组成和硫酸根含量。利用响应面优化法,依据二次回归分析确定裙带菜褐藻糖胶的最佳提取工艺为：提取温度76℃,提取时间2.5 h,料液比1︰6.5。在此条件下提取2次,粗多糖的提取率可达到7.53%。提取的粗多糖用DEAE-Sepharose Fast Flow离子交换层析分离纯化,共得到5个峰,干燥后得到5个组分。用硫酸钡比浊法测定5个组分的硫酸根含量分别为8.3%、19.2%、30.1%、38.4%和33.0%。糖腈乙酸酯衍生物气相色谱测得各组分的单糖组成分别为：组分1主要含有甘露糖和半乳糖;组分2主要含有鼠李糖,岩藻糖,木糖,甘露糖,葡萄糖和半乳糖;组分3主要含有鼠李糖,岩藻糖和甘露糖;组分4主要含有岩藻糖和半乳糖;组分5主要含有岩藻糖,甘露糖和半乳糖。     

Isolation and identification of antifungal fatty acids from the basidiomycete Gomphus floccosus

Bioautography of extracts of the fruiting bodies of the basidiomycete Gomphus floccosus (Schw.) Singer indicated the presence of fungitoxic compounds in the ethyl acetate fraction against the plant pathogens Colletotrichum fragariae, Colletotrichum gloeosporioides, and Colletotrichum acutatum. Bioassay-guided fractionation of this fraction resulted in the isolation of the bioactive fatty acids (9 S,10 E,12 Z)-9-hydroxy-10,12-octadecadienoic acid (1), (9 E,11 Z)-13-oxo-9,11-octadecadienoic acid (2), and (10 E,12 E)-9-oxo-10,12-octadecadienoic acid (3). These three oxylipins were further evaluated for activity against a greater range of fungal plant pathogens (C. fragariae, C. gloeosporioides, C. acutatum, Botrytis cinerea, Fusarium oxysporum, Phomopsis obscurans, and Phomopsis viticola) in in vitro dose-response studies. Phomopis species were the most sensitive fungi to these compounds. At 120 h of treatment, the IC50 values for compounds 1, 2, and 3 for P. obscurans were 1.0, 4.5, and 23 microM, respectively, as compared to 1.1 microM for the captan positive control.     

Effect of brown seaweed lipids on fatty acid composition and lipid hydroperoxide levels of mouse liver

Brown seaweed lipids from Undaria pinnatifida (Wakame), Sargassum horneri (Akamoku), and Cystoseira hakodatensis (Uganomoku) contained several bioactive compounds, namely, fucoxanthin, polyphenols, and omega-3 polyunsaturated fatty acids (PUFA). Fucoxanthin and polyphenol contents of Akamoku and Uganomoku lipids were higher than those of Wakame lipids, while Wakame lipids showed higher total omega-3 PUFA content than Akamoku and Uganomoku lipids. The levels of docosahexaenoic acid (DHA) and arachidonic acid (AA) in liver lipids of KK-A(y) mouse significantly increased by Akamoku and Uganomoku lipid feeding as compared with the control, but not by Wakame lipid feeding. Fucoxanthin has been reported to accelerate the bioconversion of omega-3 PUFA and omega-6 PUFA to DHA and AA, respectively. The higher hepatic DHA and AA level of mice fed Akamoku and Uganomoku lipids would be attributed to the higher content of fucoxanthin of Akamoku and Uganomoku lipids. The lipid hydroperoxide levels of the liver of mice fed brown seaweed lipids were significantly lower than those of control mice, even though total PUFA content was higher in the liver of mice fed brown seaweed lipids. This would be, at least in part, due to the antioxidant activity of fucoxanthin metabolites in the liver.     

The role of iron in peroxidation of polyunsaturated fatty acids in liposomes

This work investigated iron-catalyzed lipid oxidation in marine phospholipid liposomes. Oxygen consumption was used as a method to study lipid oxidation at pH 5.5 and 30 degrees C. The relationship between consumed oxygen and amount of peroxides (PV) and thiobarbituric reactive substances (TBARS) formed showed that both Fe2+ and Fe3+ catalyzed lipid oxidation. When Fe2+ was added to liposomes at a concentration of approximately 10 microM, an initial drop in dissolved oxygen (oxygen uptake rate >258 microM/min), followed by a slower linear oxygen uptake (oxygen uptake rate 4-6 microM/min), was observed. Addition of Fe3+ induced only the linear oxygen uptake. The initial fast drop in dissolved oxygen was due to oxidation of Fe2+ to Fe3+ by preexisting lipid peroxides (rate 79 microM Fe2+/min). Fe3+ is reduced by peroxides to Fe2+ at a slow rate (0.25 microM Fe3+/min at 30 degrees C) in a pseudo-first-order reaction. The redox cycling between Fe2+ and Fe3+ leads to an equilibrium between Fe2+ and Fe3+ resulting in a linear oxygen uptake. During the linear oxygen uptake, the interaction of Fe (3+) with lipid peroxide is the rate-limiting factor. Both alkoxy and peroxy radicals are formed by breakdown of peroxides by Fe2+ and Fe3+. These radicals react with fatty acids giving lipid radicals reacting with oxygen.     

Free polyunsaturated fatty acids cause taste deterioration of salmon during frozen storage

Increased intensity of train oil taste, bitterness, and metal taste are the most pronounced sensory changes during frozen storage of salmon (Refsgaard, H. H. F.; Brockhoff, P. B.; Jensen, B. Sensory and Chemical Changes in Farmed Atlantic Salmon (Salmo salar) during Frozen Storage. J. Agric. Food Chem. 1998a, 46, 3473-3479). Addition of each of the unsaturated fatty acids: palmitoleic acid (16:1, n - 7), linoleic acid (C18:2, n - 6), eicosapentaenoic acid (EPA; C20:5, n - 3) and docosahexaenoic acid (DHA; C22:6, n - 3) to fresh minced salmon changed the sensory perception and increased the intensity of train oil taste, bitterness, and metal taste. The added level of each fatty acid ( approximately 1 mg/g salmon meat) was equivalent to the concentration of the fatty acids determined in salmon stored as fillet at -10 degrees C for 6 months. The effect of addition of the fatty acids on the intensity of train oil taste, bitterness and metal taste was in the order: DHA > palmitoleic acid > linoleic acid > EPA. Formation of free fatty acids was inhibited by cooking the salmon meat before storage. Furthermore, no changes in phospholipid level were observed during frozen storage. The results suggest that enzymatic hydrolysis of neutral lipids plays a major role in the sensory deterioration of salmon during frozen storage.     

Effects of lactoferrin, phytic acid, and EDTA on oxidation in two food emulsions enriched with long-chain polyunsaturated fatty acids

The influence of the addition of metal chelators on oxidative stability was studied in a milk drink and in a mayonnaise system containing highly polyunsaturated lipids. Milk drinks containing 5% (w/w) of specific structured lipid were supplemented with lactoferrin (6-24 muM) and stored at 2 degrees C for up to 9 weeks. Mayonnaise samples with 16% fish oil and 64% rapeseed oil (w/w) were supplemented with either lactoferrin (8-32 muM), phytic acid (16-124 muM), or EDTA (16-64 muM) and were stored at 20 degrees C for up to 4 weeks. The effect of the metal chelators was evaluated by determination of peroxide values, secondary volatile oxidation products, and sensory analysis. Lactoferrin reduced the oxidation when added in concentrations of 12 muM in the milk drink and 8 muM in the mayonnaise, whereas it was a prooxidant at higher concentrations in both systems. In mayonnaise, EDTA was an effective metal chelator even at 16 muM, whereas phytic acid did not exert a distinct protective effect against oxidation. The differences in the equimolar effects of the metal chelators are proposed to be due to differences in their binding constants to iron and their different stabilities toward heat and low pH.     

Enzymatic production of monoacylglycerols containing polyunsaturated fatty acids through an efficient glycerolysis system

The aim of the study was to develop an efficient glycerolysis system for the enzymatic production of monoacylglycerols (MAGs) containing polyunsaturated fatty acids. Glycerolysis has been widely applied in industry for the chemical production of food MAGs under high temperature. The enzymatic glycerolysis system at 40-70 degrees C is unfortunately a multiphase system, which leads to the lower reaction efficiency. A tert-butyl alcohol system was developed after careful evaluation and more than 20-fold of the reaction efficiency from this system was obtained compared to the solvent-free system. Novozym 435 was employed as a catalyst in the glycerolysis from the screening. In the batch reaction system with tert-butyl alcohol, temperature higher than 40 degrees C was favored. The glycerol/oil ratio was best in the study with 4.5 while the solvent weight ratio from 1 to 3 had little effect. In general, 60-70% yield can be obtained at 2 h in the stirred tank reactor. The continuous glycerolysis was conducted in a packed bed reactor. MAG yield up to 70% was reached at 30-40 min residence time. The continuous glycerolysis was more sensitive to the amount of tert-butyl alcohol, and in the weight ratio to oil more than 2 was favored. The continuous process was optimized with the assistance of response surface methodology. Optimal conditions for the packed bed reactor after all considerations were recommended as glycerol/oil 4:1 (mol/mol), temperature 40 degrees C, and residence time 45 min. The operation stability study showed that there was no slight reduction of reaction performance at more than 30 days, implying a high feasibility in practical applications.     

Potential antioxidant capacity of sulfated polysaccharides from the edible marine brown seaweed Fucus vesiculosus

Fucus vesiculosus was sequentially extracted with water at 22 degrees C (fraction 1 (F1)) and 60 degrees C (F2), and with 0.1 M HCl (F3) and 2 M KOH (F4) at 37 degrees C. Soluble fractions (42.3% yield) were composed of neutral sugars (18.9-48 g/100 g), uronic acids (8.8-52.8 g/100 g), sulfate (2.4-11.5 g/100 g), small amounts of protein (< 1-6.1 g/100 g), and nondialyzable polyphenols (0.1-2.7 g/100 g). The main neutral sugars were fucose, glucose, galactose, and xylose. Infrared (IR) spectra of the fractions showed absorption bands at 820-850 and 1225-1250 cm(-1) for sulfate. F1, F2, and F4 also exhibited an absorption band at 1425 cm(-1), due to uronic acids, and their IR spectra resembled that of alginate. F3 had an IR spectrum similar to that of fucoidan with an average molecular weight of 1.6 x 10(6) Da, calculated by molecular exclusion high-performance liquid chromatography. The presence of fucose in this polysaccharide was confirmed by (1)H NMR spectroscopy. This fraction showed the highest potential to be antioxidant by the ferric reducing antioxidant power (FRAP) assay, followed by the alkali- and water-soluble fractions. Sulfated polysaccharides from edible seaweeds potentially could be used as natural antioxidants by the food industry.     

Modified egg composition to reduce low-density lipoprotein oxidizability: high monounsaturated fatty acids and antioxidants versus regular high n-6 polyunsaturated fatty acids

Eggs high in n-6 PUFA, predominant in Western markets, were found to increase blood LDL oxidation, suggesting a new health concern beyond raising cholesterol. Protective composition was explored by increasing egg antioxidants and MUFA and reducing n-6 PUFA. Lag times to plasma LDL oxidation were significantly shortened with two eggs/day of high-PUFA compositions compared to a low-egg (2-4/week) regime, by 28.8% following "HPUFA-regular" ( p < 0.01) and by 27.2% following antioxidant-fortified "HPUFA-HAOX" ( p < 0.01). However, two "HMUFA-HAOX" eggs/day with reduced egg n-6 PUFA FA% (LA by 30.7%) and PUFA:MUFA ratio (LA:OA by 45.8%) plus increased antioxidants (vitamin E 500%, carotenoids 260%), resulting in increased plasma OA 33.3%, vitamin E 22.4%, and carotenoids 55.0% ( p < 0.01), were associated with lag-time only 6.6% shorter than low-egg (NS). Among health-oriented egg modifications, here for the first time they reduced associated LDL oxidization, consistent with anti-inflammation and antioxidant paradigms, warranting further research on functional advantages of antioxidative egg composition.     

Evaluation of antioxidative activity of extracts from a brown seaweed,Sargassum siliquastrum

Antioxidative activities of the extracts from Sargassum siliquastrum were determined using the inhibition of red blood cell (RBC) hemolysis induced by 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) radicals, suppression of lipid peroxidation using rat brain homogenate, and scavenging activity of superoxide radicals. The dichloromethane fraction isolated from the methanol crude extract by differential solvent extractions exhibited the strongest antioxidant activity in both RBC hemolysis and lipid peroxidation assays. This fraction was further fractionated into four subfractions F1-F4 by silica gel column chromatography. F1 was found to be most effective in protecting RBC against AAPH radicals and in inhibiting lipid peroxidation. On the basis of thin-layer chromatography and UV and IR spectra analyses, all subfractions contained phenolic compounds. However, there was no correlation between the above antioxidant potency and total phenolic compounds estimated by using the Folin-Ciocalteau method.     

Angiotensin I-converting enzyme inhibitory peptides derived from wakame (Undaria pinnatifida) and their antihypertensive effect in spontaneously hypertensive rats

Seven kinds of angiotensin I-converting enzyme (ACE) inhibitory peptides were isolated from the hydrolysates of wakame (Undaria pinnatifida) by Protease S "Amano" (from Bacillus stearothermophilus) by using three-step high-performance liquid chromatography (HPLC) on a reverse-phase column. These peptides were identified by amino acid composition analysis, sequence analysis, and liquid chromatography-mass spectrometry (LC-MS), as Val-Tyr (IC(50) = 35.2 microM), Ile-Tyr (6.1 microM), Ala-Trp (18.8 microM), Phe-Tyr (42.3 microM), Val-Trp (3.3 microM), Ile-Trp (1.5 microM), and Leu-Trp (23.6 microM). These peptides have resistance against gastrointestinal proteases in vitro. Each peptide was determined to have an antihypertensive effect after a single oral administration in spontaneously hypertensive rats (SHR). Among them, the blood pressure significantly decreased by Val-Tyr, Ile-Tyr, Phe-Tyr, and Ile-Trp in a dose of 1 mg/kg of body weight (BW). The present study showed that antihypertensive effect in the hydrolysates of wakame by Protease S "Amano" was attributed to these peptides.     

Influence of dissolved gases and heat treatments on the oxidative degradation of polyunsaturated fatty acids enriched dairy beverage

The combined effect of dissolved gas composition and heat treatment on the oxidative degradation of a dairy beverage enriched with 2% linseed oil was studied. The dairy beverage was saturated with air, nitrogen, or a nitrogen/hydrogen mixture (4% hydrogen) before pasteurization or sterilization. Saturation with either nitrogen or a nitrogen/hydrogen mixture decreased the dissolved oxygen concentration in dairy beverages (Delta = 7.7 ppm), and the presence of hydrogen significantly reduced the redox potential (Delta = 287 mV). Heat treatments also reduced the oxygen content and redox potential, sterilization being more effective than pasteurization. Both pasteurization and sterilization induced the oxidative degradation of the beverages. On average, the propanal concentration increased by a factor of 2.3 after pasteurization and by a factor of 6.2 after sterilization. However, during storage, sterilized beverages resisted light-induced oxidation better than unheated or pasteurized beverages. Furthermore, saturation with nitrogen or a nitrogen/hydrogen mixture significantly reduced oxidative degradation and provided some protection against color changes during storage.     

Effect of lipid composition on meat-like model systems containing cysteine, ribose, and polyunsaturated fatty acids

This paper compares the volatile constituents of model systems containing the important meat aroma precursors cysteine and ribose, with and without either methyl linoleate, an n-6 fatty acid, or methyl alpha-linolenate, an n-3 acid, both of which are present in meat. Many of the volatile compounds formed from the reaction between cysteine and ribose were not formed, or formed in lower amounts, when lipid was present. This may be due to the reaction between hydrogen sulfide, formed from the breakdown of cysteine, and lipid degradation products. In addition, cysteine and ribose modified lipid oxidation pathways, so that alcohols and alkylfurans were formed rather than saturated and unsaturated aldehydes. Several volatile compounds, which have been found at elevated levels in cooked meat from animals fed supplements high in n-3 acids, were formed when methyl alpha-linolenate reacted with cysteine and ribose. The possible effects of increasing the n-3 content of meat upon flavor formation during cooking are discussed.     

Determination of cis, cis-methylene interrupted polyunsaturated fatty acids in fats and oils by capillary gas chromatography

A capillary gas chromatographic (CGC) method is described for the determination of cis,cis-methylene interrupted polyunsaturated fatty acids (cis-PUFA) in fats and oils. The sample is saponified and the liberated fatty acids are esterified to the corresponding methyl esters. The latter are analyzed by CGC using a 60 M. SP2340 capillary column. Area percent values for 9,12-cis,cis-C18:2 and 9,12,15-cis,cis,cis-C18:3 fatty acid methyl esters are summed to give the total cis-PUFA content. Gas chromatographic results agreed well with those obtained by an enzymatic lipoxygenase method at the 31-48% cis-PUFA levels with a correlation coefficient of 0.98. The method has a precision (relative standard deviation) of 0.33% at a 44.4% cis-PUFA level in margarine oil.     

Oxidative stability of fish and algae oils containing long-chain polyunsaturated fatty acids in bulk and in oil-in-water emulsions

The oxidative stability of long-chain polyunsaturated fatty acid (PUFA) and docosahexaenoic acid (DHA)-containing fish and algae oils varies widely according to their fatty acid composition, the physical and colloidal states of the lipids, the contents of tocopherols and other antioxidants, and the presence and activity of transition metals. Fish and algal oils were initially much more stable to oxidation in bulk systems than in the corresponding oil-in-water emulsions. The oxidative stability of emulsions cannot, therefore, be predicted on the basis of stability data obtained with bulk long-chain PUFA-containing fish oils and DHA-containing algal oils. The relatively high oxidative stability of an algal oil containing 42% DHA was completely lost after chromatographic purification to remove tocopherols and other antioxidants. Therefore, this evidence does not support the claim that DHA-rich oils from algae are unusually stable to oxidation. Addition of ethylenediaminetetraacetic acid (EDTA) prevented oxidation of both fish and algal oil emulsions without added iron and at low iron:EDTA molar concentrations. EDTA, however, promoted the oxidation of the corresponding emulsions that contained high iron:EDTA ratios. Therefore, to be effective as a metal chelator, EDTA must be added at molar concentrations higher than that of iron to inhibit oxidation of foods containing long-chain PUFA from either fish or algae and fortified with iron.     

Isolation and characterization of two flavonoids, engeletin and astilbin, from the leaves of Engelhardia roxburghiana and their potential anti-inflammatory properties

Engeletin, a flavonoid compound, was isolated from the leaves of Engelhardia roxburghiana for the first time, along with astilbin, another flavonoid. The chemical structures of engeletin and astilbin were confirmed by (1)H and (13)C nuclear magnetic resonance (NMR) and mass spectrometry (MS) spectra, and their anti-inflammatory activities were studied in lipopolysaccharide (LPS)-stimulated mouse J774A.1 macrophage cells. LPS induced the inflammatory state in macrophage cells and increased mRNA expressions of pro-inflammatory cytokines. Engeletin and astilbin exhibited remarkable inhibitory effects on interleukin (IL)-1β and IL-6 mRNA expression. Significant inhibition of LPS-mediated mRNA expressions were also seen in LPS binding toll-like receptor (TLR)-4, pro-inflammatory cytokine tumor necrosis factor (TNF)-α, IL-10, chemoattractant monocyte chemotactic protein (MCP)-1, and cyclooxygenase (COX)-2 genes. The reduced expression of these cytokines may alleviate immune response and reduce inflammatory activation, indicating that engeletin and astilbin may serve as potential anti-inflammatory agents.     

Profile of trans fatty acids (FAs) including trans polyunsaturated FAs in representative fast food samples

The content of trans fat in foods is most commonly determined by summing the levels of individual trans fatty acids (FAs), analyzed as FA methyl esters (FAME) by gas chromatography. Current Official Methods of the American Oil Chemists' Society (AOCS) enable quantitation of total trans fat in foods but were not designed for the determination of transFA isomeric compositions. In the present study, the content of trans fat in 32 representative fast food samples ranged from 0.1 to 3.1 g per serving, as determined according to AOCS Official Method Ce 1j-07. Further analysis of FAME using the 200 m SLB-IL111 ionic liquid column yielded quantitative results of total, trans, saturated, and cis unsaturated fat that were comparable to those of Method Ce 1j-07 and also allowed for the complementary determination of individual trans 18:1, trans 18:2, and trans 18:3 FA isomeric compositions under conditions suitable for routine sample analysis.     

Effects of vitamin E supplementation on lipid peroxidation and color retention of salted calf muscle from a diet rich in polyunsaturated fatty acids.

The color of fresh meat is one of the most important quality criteria of raw muscle foods. This red color is principally due to the presence of oxymyoglobin. The present study was undertaken to examine the effect of a diet rich in polyunsaturated fatty acids (PUFA), the addition of NaCl, and the influence of dietary supplementation with vitamin E on calf muscle oxymyoglobin oxidation (color) and lipid peroxidation. Vitamin E was added to the feed at a concentration of 4000 mg/day for 90 days before slaughter. This diet increased the alpha-tocopherol concentration in muscle membrane from 2.6-2.8 to 6.5-7.0 microg/g of fresh weight. It was found that the diet rich in PUFA and, especially, the addition of NaCl increased muscle lipid peroxidation and oxymyoglobin oxidation as indicated by the contents of thiobarbituric acid-reactive substances and substances that impaired color value readings during storage at 4 degrees C. Both undesirable reactions during storage were controlled very efficiently by the presence of a critically high concentration of alpha-tocopherol in the muscle tissues. The findings concerning the antioxidant activity of alpha-tocopherol in this study form additional evidence of its efficient protection against oxidative reactions during storage of muscle tissues and its potential to maintain a high nutritional value in them.     

Insecticidal fatty acids and triglycerides from Dirca palustris.

Five compounds, 1-5, were isolated from the seed hexane extract of Dirca palustris. Compounds 1-3 were triglycerides, and 4 and 5 were linoleic and oleic acids, respectively. Compounds 1-3 were not biologically active; however, 4 (linoleic acid) and 5 (oleic acid) were insecticidal against fourth instar Aedes aegyptii larvae and exhibited potent feeding deterrent activity against neonate larvae of Helicoverpa zea, Lymantria dispar, Orgyia leucostigma, and Malacosoma disstria.     

Primary aminophospholipids in the external layer of liposomes protect their component polyunsaturated fatty acids from 2,2'-azobis(2-amidinopropane)- dihydrochloride-mediated lipid peroxidation

We showed in our previous study that docosahexaenoic acid-rich phosphatidylethanolamine in the external layer of small-size liposomes, as a model for biomembranes, protected its docosahexaenoic acid from 2,2'-azobis(2-amidinopropane)dihydrochloride- (AAPH-) mediated lipid peroxidation in vitro. Besides phosphatidylethanolamine, both phosphatidylserine and an alkenyl-acyl analogue of phosphatidylethanolamine, phosphatidylethanolamine plasmalogen, are reported to possess characteristic antioxidant activities. However, there are few reports about the relationship between the protective activity of phosphatidylethanolamine plasmalogen and/or phosphatidylserine against lipid peroxidation and their distribution in a phospholipid bilayer. Furthermore, it is unclear whether phosphatidylethanolamine plasmalogen and/or phosphatidylserine protect their component polyunsaturated fatty acids (PUFAs) from lipid peroxidation. In the present study, we examined the relationship between the transbilayer distribution of aminophospholipids, such as phosphatidylethanolamine rich in arachidonic acid, phosphatidylethanolamine plasmalogen, and phosphatidylserine, and the oxidative stability of their component PUFAs. The transbilayer distribution of these aminophospholipids in liposomes was modulated by coexisting phosphatidylcholine bearing two types of acyl chain: dipalmitoyl or dioleoyl. The amounts of these primary aminophospholipids in the external layer became significantly higher in liposomes containing dioleoylphosphatidylcholine than in those containing dipalmitoylphosphatidylcholine. Phosphatidylethanolamine rich in arachidonic acid, phosphatidylethanolamine plasmalogen or phosphatidylserine in the external layer of liposomes, as well as external docosahexaenoic acid-rich phosphatidylethanolamine, were able to protect their component PUFAs from AAPH-mediated lipid peroxidation.