Improved approach for analyzing bromophenols in seafood using stable isotope dilution analysis in combination with SPME

An analytical method for the measurement of five naturally occurring bromophenols of sensory relevance in seafood (barramundi and prawns) is presented. The method combines simultaneous distillation-extraction followed by alkaline back extraction of a hexane extract and subsequent acetylation of the bromophenols. Analysis of the bromophenol acetates was accomplished by headspace solid phase microextraction and gas chromatography-mass spectrometry using selected ion monitoring. The addition of (13)C 6 bromophenol stable isotope internal standards for each of the five congeners studied permitted the accurate quantitation of 2-bromophenol, 4-bromophenol, 2,6-dibromophenol, 2,4-dibromophenol, and 2,4,6-tribromophenol down to a limit of quantification of 0.05 ng/g of fish flesh. The method indicated acceptable precision and repeatability and excellent linearity over the typical concentration range of these compounds in seafood (0.5-50 ng/g). The analytical method was applied to determine the concentration of bromophenols in a range of farmed and wild barramundi and prawns and was also used to monitor bromophenol uptake in a pilot feeding trial.     

Distribution of bromophenols in species of marine algae from eastern Australia

Forty-nine species (87 samples) of marine macroalgae from eastern Australia were analyzed by GC/MS for the key seafood flavor components 2- and 4-bromophenol, 2,4- and 2,6-dibromophenol, and 2,4, 6-tribromophenol. All five bromophenols were found in 62% of samples, four in 32% of samples, and three in the remaining 6% of samples. 2, 4,6-Tribromophenol was found in all samples and, with few exceptions, was present in the highest concentrations. The total bromophenol content determined on a wet-weight basis varied widely across species from 0.9 ng/g in the green alga Codium fragile to 2590 ng/g in the red alga Pterocladiella capillacea. Species with the highest concentrations of bromophenols were all collected from sites exposed at low tide. The study demonstrates the wide occurrence of bromophenols in marine algae and provides a possible source of such compounds in fish that feed predominantly on ocean plants. The possible effect that dietary marine algae has on the flavor of omnivorous ocean fish is discussed.     

Seasonal distribution of bromophenols in selected Hong Kong seafood

Selected seafood including rabbitfish (Siganus canaliculatus), brown-spotted grouper (Epinephelus areolatus), clam (Tapes philippinarum), oyster (Ostrea rivularis), shrimp (Penaeus japonicus), and crab (Charybdis feriatus), commonly found in the Hong Kong wet market, was monitored for their distribution and seasonal variations of their bromophenol contents. Specifically, 2-bromophenol (1). 4-bromophenol (2). 2,4-dibromophenol (3). 2,6-dibromophenol (4). and 2,4,6-tribromophenol (5). were monitored due to their flavor impact to seafood. All samples of marine origin contained bromophenols throughout a year. Crab had the highest concentration of total bromophenol content throughout the season. Concentrations of compounds 1, 4, and 5 in the local seafood were generally higher than that found in the literature values to provide characteristic flavor, but lower than that required to cause off-flavor. Variations of the flavor impact of bromophenols in seafood during a season could be better shown by their flavor values. Distribution and seasonal variations of bromophenol content in seafood coincided well with the seasonal growth cycle of the bromophenol synthesizing seaweeds, e.g. brown algae, in the region suggesting the abundant source of bromophenols in the environment might have a high impact on the quantity of bromophenols found in seafood.     

Distribution of bromophenols in species of marine polychaetes and bryozoans from eastern Australia and the role of such animals in the flavor of edible ocean fish and prawns (shrimp).

Sixteen species (44 samples) of marine polychaetes and 10 species (14 samples) of bryozoans from eastern Australia were analyzed by GC/MS for the key seafood flavor components 2- and 4-bromophenol, 2, 4- and 2,6-dibromophenol, and 2,4,6-tribromophenol. All five bromophenols were found in 91% of polychaetes and 64% of bryozoans. The remaining samples all contained at least three bromophenols. 2,4, 6-Tribromophenol was found in all polychaetes and bryozoans and, with few exceptions, was present in the highest concentrations. The total bromophenol content determined on a wet-weight basis varied widely between species: for polychaetes, from 58 ng/g for Australonuphis teres to 8.3 million ng/g for Barantolla lepte, and for bryozoans, from 36 ng/g for Cladostephus spongiosus to 1668 ng/g for Amathia wilsoni. Species of polychaetes with the highest concentrations of bromophenols were all collected from muddy environments. The possible effects that dietary polychaetes and bryozoans have on the ocean-, brine-, or iodine-like flavors of certain seafoods are discussed.     

Seasonal variations of bromophenols in brown algae (Padina arborescens,Sargassum siliquastrum,and Lobophora variegata) collected in Hong Kong

Distributions and seasonal variations of the key seafood flavor compounds including 2-bromophenol, 4-bromophenol, 2,4-dibromophenol, 2,6-dibromophenol, and 2,4,6-tribromophenol in three species of brown algae (Padina arborescens, Sargassum siliquastrum, and Lobophora variegata) found in Hong Kong waters were investigated. Bromophenols were extracted by simultaneous steam distillation and solvent extraction apparatus and analyzed by gas chromatography-mass spectrometry. On a dried weight basis, the total bromophenol content (TBC) determined varied widely with seasons (from 40.9 to 7030 ng/g). The TBCs detected were higher in winter and lower in summer. Except for 2-bromophenol, the rest of the bromophenols were detected in all of the algal samples. The TBC of L. variegata was generally the highest among all of the algae collected. Relatively high concentrations of bromophenols in algae supported the fact that marine algae were major producers of bromophenols in the marine environment.     

Determinations of residual furazolidone and its metabolite, 3-amino-2-oxazolidinone (AOZ), in fish feeds by HPLC-UV and LC-MS/MS, respectively

The antibacterial drug furazolidone belonging to the group of nitrofuran antibacterial agents has been widely used as an antibacterial and antiprotozoal feed additive for poultry, cattle, and farmed fish in China. During application a large proportion of the administered drug may reach the environment directly or via feces. Although the use of furazolidone is prohibited in numerous countries, there are indications of its illegal use. It is known that furazolidone can be rapidly metabolized to 3-amino-2-oxazolidinone (AOZ) in the body of the target organism. In this study, a total of 21 fish feed samples, including 17 commercial fish feeds from local markets in China (representing 15 different formulations) and 4 fish feeds obtained from Germany and Turkey, respectively, are analyzed to determine whether the drug is still illegally used or commercially available feeds are contaminated by this drug. High-performance liquid chromatography (HPLC) and liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) methods have been implemented to determine furazolidone and its metabolite AOZ in fish feeds containing animal protein, respectively. An efficient and convenient cleanup method for the determination of furazolidone in fish feeds is developed, and a simple cleanup method for the determination of AOZ is used. Method recoveries for samples used were determined as 87.7-98.3% for furazolidone at two spike levels of 2.0 and 5.0 ng g-1 and as 95.6-102.8% for AOZ at spike levels of 0.4 and 0.8 ng g-1. Limits of detections were 0.4 ng g-1 for furazolidone and 0.05 ng g-1 for AOZ. The established methods are therefore suitable for the determination of furazolidone and its metabolite AOZ in fish feeds at trace contamination levels. Using the established methods, all fish feed samples have been proved to be furazolidone negative; however, AOZ is tested in 16 of 17 fish feeds obtained from local markets in the Hubei province of China, with a positive rate as high as 94.1%.     

Determination of phosphorus fractions in animal protein ingredients

Phosphorus (P) is present in different chemical compounds in animal feeds, and the solubility and digestibility of these different compounds are known to differ significantly. Animal protein ingredients generally have a high P content and are major contributors to total P of feeds for fish and other domestic animals. Estimation of different P compounds in these ingredients could help to improve the accuracy of estimates of digestible P contents of feeds. Bone P and organic P contents were quantified in 32 animal protein ingredients, including 10 fish meals, 14 meat and bone meals, and 8 poultry byproducts meals, using a fractionation protocol. The total P contents of the ingredients ranged from 2.1 to 8.3% on a dry matter (DM) basis. Organic P contents varied between 0.3 and 1.3% of DM. Highly significant (p < 0.001) linear relationships were observed between total P and ash and between bone P and ash for all ingredients combined: total P (%) = 0.185 x ash (%) (R (2) = 0.88), and bone P (%) = 0.188 x ash (%) - 0.852 (R (2) = 0.94). These results suggest that bone P can be easily and reliably estimated on the basis of ash content in animal protein ingredients.     

Twin‐Screw Extrusion Processing of Rainbow Trout (Oncorhynchus mykiss) Feeds Using Various Levels of Corn‐Based Distillers Dried Grains with Solubles (DDGS)

Increasing demand for seafood products and rising demand for fish meal for commercial fish feeds is driving the search for effective alternative protein sources. Twin‐screw extrusion trials were conducted to study the production of nutritionally balanced feeds for rainbow trout fingerlings (Oncorhynchus mykiss). Six isocaloric (≈4.61 kcal/g) ingredient blends with a target protein content of >45% db were formulated with 0, 10, 20, 30, 40, and 50% distillers dried grains with solubles (DDGS) and other feed ingredients. The moisture contents of the diets were initially adjusted to 5–7% db, and then extruded at 250 rpm using dual 1.9 mm dies with varying amounts of steam (7.2–7.7 kg/hr) injected into the conditioner and water (4.3–6.5 kg/hr) into the extruder. Mass flow rates, moisture contents, and temperatures were measured during processing and moisture content, water activity, unit density, bulk density, expansion ratio, compressive strength, compressive modulus, pellet durability index, water stability, and color were analyzed to quantify the effects of varying DDGS content on the extrudate physical properties. Significant differences (P < 0.05) among the blends were observed for color and bulk density for both the raw and extruded materials, respectively, and for the unit density and pellet durability index of the extruded products. There were also significant changes in redness and yellowness, but only minor changes in brightness, among the final products with increasing DDGS content. The compressive strength of the extrudates increased significantly with increasing DDGS. Expansion ratio of all pellets was low. All extruded diets achieved very good water stability.     

高含量乳清粉的仔猪配合饲料热特性及调质温度控制

为探究热敏性饲料原料乳清粉及不同含量乳清粉的仔猪配合饲料的热物理特性,该文以仔猪料配方中的4种主要饲料原料玉米、豆粕、乳清粉和鱼粉为研究对象,采用混料设计的方法得到33种不同含量(0～30％)乳清粉的仔猪配合饲料,并利用差示扫描量热法(differential scanning calorimetry,DSC)测定了4种单一原料在25～120℃范围内以及33种仔猪配合饲料在25～110℃范围内的比热,分析了乳清粉及高含量乳清粉(质量分数≥14.548％)的仔猪配合饲料的热变性过程.结果显示:玉米、豆粕和鱼粉的比热分别与温度(25～120℃)呈线性、对数和二次关系,而乳清粉的比热与温度(25～110℃)遵循三次多项式的关系;当配合饲料中含有≥6.25％的乳清粉时,其比热与温度遵循三次多项式的关系;配合饲料的比热显著受温度、原料配比以及二者交互作用的影响(P＜0.001),其中,温度的影响最为显著,而乳清粉含量的影响次之.DSC热焓曲线上,乳清粉在109.79℃会出现吸热峰,为乳清蛋白的热变性导致;而随着温度由20℃升高到110℃,乳清粉颗粒由存在许多凸起与微孔的粗糙表面结构逐渐过渡为光滑、粘结的状态.与乳清粉相似,高含量乳清粉的配合饲料也会在77.95～87.69℃出现吸热峰.在仔猪配合颗粒饲料的加工过程中,为降低乳清蛋白的变性程度、减少环模制粒机的堵机现象,应将调质温度降低至70℃以下为宜.研究结果为高含量乳清粉的仔猪配合饲料的调质、制粒等热处理过程的工艺优化提供理论指导.     

Qualitative and quantitative liquid chromatographic analysis methods for the determination of the effects of feed supplements on hen egg yolk phospholipids

Quantitative and qualitative high-performance liquid chromatographic methods were utilized to separate phospholipid classes. After qualitative separation, the fatty acid moieties of each separated phospholipid class were determined using a gas chromatographic method. On the basis of these analyses, the effect of supplemented feeds on hen egg yolk lipids can be evaluated. The supplemented feeds contained 1-5% of vegetable-based or fish oils. The phospholipid content and composition were the same in all feeding groups, the proportions of phosphatidylcholines, phosphatidylethanolamines, and sphingomyelins being 70, 28, and 3%, respectively. In each feeding group, the fatty acid profiles of phosphatidylcholines and sphingomyelins were similar to each other and different from that of phosphatidylethanolamines. The supplemented feeds had a statistically significant (p < 0.05) effect on the fatty acid composition of phosphatidylcholines. The supplements decreased the proportion of saturated fatty acids in total fat, but this effect was not found in phospholipids.     

无水保活条件下团头鲂生理应激及鱼肉品质的变化

为探讨无水保活对团头鲂生理应激和肌肉品质的影响,该文研究了团头鲂无水保活所需的较佳条件,测定了未处理(对照)、无水和恢复3种状态下团头鲂的血液生化以及鱼肉成分、持水力、新鲜度、质构和游离氨基酸等品质指标。结果表明:团头鲂无水保活的较佳生活水温为(14.8±0.2)℃(冬季),以2℃/h降至休眠温度2℃,充氧包装后在2℃无水保活14 h,在当季暂养水温中((14.8±0.2)℃)恢复24 h后存活率为100%;无水保活14 h后,血液中皮质醇和乳酸浓度、谷草转氨酶和谷丙转氨酶活力显著高于未处理(对照)(P0.05),血糖浓度和碱性磷酸酶活性明显低于未处理值(P0.05),当季暂养水温恢复24 h后,基本都恢复至未处理组值,但谷草转氨酶仍明显高于未处理组值(P0.05);低温无水环境对团头鲂鱼肉中蛋白质和脂肪含量、新鲜度和持水力无影响,但鱼肉的硬度和咀嚼性明显下降(P0.05),鱼肉游离氨基酸中丝氨酸(Ser)、谷氨酸(Glu)、甘氨酸(Gly)、缬氨酸(Val)和精氨酸(Arg)等含量显著升高(P0.05),异亮氨酸(Ile)、亮氨酸(Leu)、酪氨酸(Tyr)和苯丙氨酸(Phe)在无水保活过程中无明显变化,当季暂养水温恢复后却显著高于未处理组水平(P0.05)。因此,团头鲂无水保活中产生的应激反应在恢复24 h后基本消除,且无水保活对鱼肉的食用品质影响较小。该研究结果为无水保活在淡水鱼活鱼运输中的应用提供参考。     

黄颡鱼的含肉率及肌肉营养价值研究

采用常规方法测得广西桂江黄颡鱼的含肉率为 83.88% ;每 1 0 0 g肌肉 (干样 )中含粗蛋白85.96g,粗脂肪 5.0 4 g,粗灰分 5.57g,氨基酸总量为 70 .2 1 g,必需氨基酸含量 2 7.39g,含钙 39.58mg,磷 1 87mg,铜 0 .33mg,锌 5.33mg,铁 4 .75mg,锰 3.0 3mg,硒 74 7.0 6μg。综合分析结果认为 ,广西桂江的黄颡鱼是营养价值较高的优质鱼类     

Determination of folate concentrations in diverse potato germplasm using a trienzyme extraction and a microbiological assay

Folate deficiency is a leading cause of birth defects and is implicated in several other diseases. We are interested in how much folate concentrations vary among potato germplasm. We determined total folate concentrations of potato tubers from 67 cultivars, advanced breeding lines, or wild species. Folates were extracted by a tri-enzyme treatment and analyzed by using a Lactobacillus rhamnosus microbiological assay. Folate concentrations varied from 521 +/- 96 to 1373 +/- 230 ng/g dry weight and were genotype and location dependent. The highest folate concentrations were mostly found in color-fleshed potatoes. Variations of folate concentrations within either color- or white-fleshed tubers were similar ( approximately 2-fold). Skin contained approximately 30% higher folate concentrations than flesh. Storage of tubers for 7 months generally led to an increase in folate contents. Semiquantitative RT-PCR analyses showed that higher folate contents were correlated with lower mRNA expression of some folate genes.     

Metabolism of chlorophenols by Rhodotorula glutinis

A phenol-utilizing strain of Rhodotorula glutinis was isolated from Rothamsted soil. Washed, phenol-grown R. glutinis cells oxidized 3- and 4-chlorophenols to 4-chlorocatechol, and 4-bromophenol presumably to 4-bromocatechol. Phenol-grown cells consumed O₂ in the presence of 2-, 3- or 4-chlorophenol, 4-bromophenol, 2,4-dichloro- and 2,4-dibromophenol. Benzoate-grown cells showed O₂-uptakes in the presence of 2-, 3- or 4-chlorobenzoate but the formation of catechol compounds was not definitely established.     

Survey of dioxin-like compounds in dairy feeds in the United States

The United States Environmental Protection Agency (USEPA) has completed a survey of dioxin-like compounds (including 17 dioxin and furan (CDD/F) congeners and 12 coplanar polychlorinated biphenyl (PCBs) congeners) in dairy feeds from 10 dairy research facilities around the United States, sampling the overall mixtures and the major and minor feed components. Low levels of dioxin were found in all feed mixtures with an average concentration of 0.05 pg/g (ppt) toxic equivalent (TEQ) dry weight. This is lower than previously found in dairy feeds by about a factor of 4. While it is possible that generally lower levels of dioxins in the environment in recent years may explain this result, examinations of the data suggest that the oven drying used to prepare the wet feed samples could have resulted in a loss of dioxins from the feed materials. The percentage of the total TEQ due to CDD/Fs was about four times that of PCBs. Leafy vegetations in the feed (the silages and the hays) had concentrations about twice as high as nonleafy, protected vegetation of the feeds (the ground or meal corn, cottonseed, and others). Minor components did not significantly influence the final feed mixture concentration of dioxin TEQ. However, in one of the feed mixtures, a minor component with a high concentration of 38.5 ppt TEQ effectively doubled the concentration of the overall feed mixture.     

冷熏对高白鲑理化性质及肌球蛋白构象的影响

以新疆塞里木湖高白鲑为研究对象,研究冷烟熏过程对鱼肉理化性质及肌球蛋白构象的影响。冷熏温度为(20±2)℃,发烟温度140℃,冷熏时间分别为0、6、12、18、24 h,研究冷熏鱼肉理化性质(含水率、水分活度、色泽、质构特性、TVB-N)及肌球蛋白构象(蛋白浓度、总巯基含量、Ca^2+-ATPase、表面疏水性)变化。研究结果表明,冷熏0~24h,鱼肉含水率由75.75%下降至53.03%,水分活度(Aw)由0.988下降至0.952;鱼肉亮度值(L^*)显著降低(P<0.05),红度值(a^*)缓慢增加,黄度值(b^*)显著增加(P<0.05);表征鱼肉质构特性的剪切力与韧性均呈明显上升趋势(P<0.05);鱼肉挥发性盐基氮(TVB-N)由9.81上升至14.23 mg/100g;肌球蛋白浓度、总巯基含量、Ca^2+-ATPase活性均显著降低(P<0.05),表面疏水性增加(P<0.05);综上,控制冷熏时间12~18 h有利于提高冷熏鱼肉品质,降低鱼肉肌球蛋白变性与氧化程度。该研究为特色淡水鱼冷熏制品开发与烟熏过程中蛋白氧化调控提供理论依据与技术参考。     

叶面喷施钾、钙和镁肥调节三月红荔枝果皮着色和果肉风味变化同步的效果

以三月红荔枝(Litchi chinensis Sonn.cv Sanyuehong)成年树为试材,以叶面喷施0.5%KCl(K)、0.3%Ca Cl2(Ca)和1.5%Mg Cl2(Mg)以及它们的混合液为处理,以清水为对照(CK),探讨叶面喷施K、Ca和Mg肥对三月红荔枝果皮着色和果肉风味变化同步的调节效果。结果表明,果实在生长发育过程中,各处理和CK的果皮色角度(h)值和果肉总酸含量在总体上呈下降趋势,果肉可溶性糖含量和果肉糖酸比在总体上呈上升趋势。不同处理的果肉糖酸比动态变化趋势在后期存在差异,即从4月27日到30日,Ca和Mg处理无显著变化,其余处理和CK均显著下降,且Ca处理显著高于CK;不同处理的果皮h值动态变化趋势也在后期存在差异,即Ca+Mg处理在4月27日降至最低并显著低于CK,至4月30日,其一直无显著变化,但其余处理在此期间则均显著下降,且Ca处理在4月30日显著低于其它处理和CK。可见,Ca+Mg、Ca和Mg处理的果皮h值与果肉糖酸比均同步达到最佳,其中Ca+Mg和Ca处理同时改善了果皮着色和果肉风味品质。     

Effect of Waterborne Zinc on Survival, Growth, and Feed Intake of Indian Major Carp, Cirrhinus mrigala (Hamilton)

The effect of waterborne zinc on survival, growth, and feed intake of Indian major carp, Cirrhinus mrigala (Hamilton), advanced fry was studied under laboratory condition. Survival rates of C. mrigala advanced fry (2.71?±?0.49 g) after 30 days exposure to control (0.01), 0.03, 0.06, 0.10, and 0.15 mg/L zinc using the static renewal method in freshwater at pH 7.3?±?0.2, temperature 26?±?2°C, and total hardness 114?±?16 mg/L as CaCO₃ were 100%. Growth of the fish exposed to 0.10 and 0.15 mg/L of zinc was significantly lower (P?<?0.05) than in control (0.01), 0.03, and 0.06 mg/L of zinc after 30 days of exposure. However, there were no significant differences (P?>?0.05) in fish growth between 0.03 and 0.06 mg/L zinc concentrations. Feed intake rates were significantly (P?<?0.05) reduced in the fish exposed to 0.10 mg/L and higher levels of zinc. The zinc accumulation in the whole body of the fish increased with increasing concentrations of the metal.     

Collagen content in farmed Atlantic salmon (Salmo salar L.)

To clarify fish flesh quality problems and softening of fish muscle tissue during chilled storage, the collagen content, types I and V, and its changes in solubility during storage on ice in muscle of farmed Atlantic salmon (Salmo salar L.) were analyzed. The contents of acid-soluble, pepsin-soluble, and insoluble collagen in white muscle were determined in fresh fish muscle and after 3 days of storage in ice. The total collagen content was 2.9 g kg(-)(1) fresh weight. During storage on ice, a progressive change in solubility of muscle collagen was found. For acid- and pepsin-soluble collagen fractions, significantly higher and lower values, respectively, were detected. The presence and quantification of types I and V collagen in the different collagen fractions was determined also, but no significant difference in solubility during storage was found. The result suggested that collagen fibers of Atlantic salmon have a high solubility in acid solutions and contain few cross-links.     

喷施钙对肥城桃果活性钙含量及其在亚细胞分布的影响

【目的】肥城桃是山东特产,但经常发生缝合线褐变、 不耐储藏等问题,补充钙肥是减轻其生理病害的有效措施。了解肥城桃的需钙规律,为肥城桃补充钙素营养提供理论依据和技术指导。【方法】以11年生红里肥桃为试材,从花后一个月开始,每隔30 d在选取的肥桃果树上喷施钙肥。试验设3个处理: 1)喷施0.5%氨基酸钙溶液; 2)喷施0.5%硝酸钙; 3)喷施清水为对照。从4月28日起每隔30 d采样,测定果皮、 果肉、 果核、 果仁总钙含量及果肉钙组分含量,在成熟期取果实用透射电镜观察果肉细胞内钙的亚细胞分布。【结果】3个处理果实中的钙含量均以幼果期最高,随着果实的成熟,全钙、 水溶性钙、 果胶酸钙含量均呈下降趋势,喷钙处理在一定程度上提高了果皮、 果肉、 果核及果仁的总钙含量,其中果肉总钙含量变化最明显,在果实成熟期,喷施氨基酸钙及硝酸钙的处理果肉总钙含量分别增加了68%、 77%。通过电镜观察,喷钙果肉细胞中钙均匀分布于细胞壁、 细胞膜、 液泡膜上,液泡中有钙的堆积; 未喷钙细胞壁中钙的分布减少,细胞膜、 液泡膜上钙也均匀分布; 发生褐变的果肉细胞内钙分布很少且不均匀。【结论】肥城桃果实中全钙含量随着果实生长而迅速下降。喷施钙肥能提高果实全钙尤其是水溶性钙及果胶钙的含量,增加细胞壁钙的分布,有利于缓解果实发育过程中钙含量的下降。喷施氨基酸钙和硝酸钙都能增加肥城桃果肉的不溶性果胶含量,提高果实硬度。