The Condensed Tannin Content of a Range of Subtropical and Temperate Forages and the Reactivity of Condensed Tannin with Ribulose- 1,5-bis-phosphate Carboxylase (Rubisco) Protein

A series of subtropical grasses and temperate grasses, herbs and legumes were analysed for the presence of extractable and bound condensed tannin (CT) using colorimetric analysis by the butanol–HCl method. Condensed tannins are routinely purified using affinity chromatography with Sephadex LH-20 as a matrix. Therefore, Sephadex LH-20 extracts were further analysed for the presence of CT by ¹³C nuclear magnetic resonance, for anthocyanidin formation after butanol–HCl treatment and for their ability to precipitate ribulose-1,5-bisphosphate carboxylase (Rubisco) protein from lucerne, at pH 7·0. Criteria for the presence or absence of CT were defined. Trace amounts of CT (0·2–2·5 g kg⁻¹ dry matter; DM) were identified and confirmed in summer grass (Digiteria sanguinalis), perennial ryegrass (Lolium perenne) and red clover (Trifolium pretense), with chicory (Chicorium intybus), lucerne (Medicago sativa) and plantain (Plantago lanceolata) identified as probably containing CT. It was concluded that the subtropical grasses kikuyu (Pennisetum clandestinum), paspalum (Paspalum diatatum), smooth witchgrass (Panicum dichotomiflorum) and crowfoot (Eleusine indica) and the temperate grass, Yorkshire fog (Holcus lanatus) probably did not contain CT. Analysis of the extractable fractions by vanillin–HCl gave higher values for CT than analysis by butanol–HCl and wrongly identified some forages as containing trace levels of CT. It was concluded that vanillin–HCl was not specific enough for the detection of trace levels of CT in forages. These results raise the possibility of plant selection programmes to increase the level of CT in grazed forages to approximately 5 g kg⁻¹ DM, the suggested minimum level required to prevent bloat in cattle and to increase wool growth in sheep. It is suggested that this be considered for perennial ryegrass, chicory, red clover and lucerne.     

Trypsin Inhibitor Activity and Tannin Content Do Not Affect Calcium Bioavailability of Three Commonly Consumed Legumes

Calcium bioavailability from legumes containing a range in trypsin inhibitor activity (TIA) and tannin concentration was studied. Three cultivars (Phaseolus vulgaris) were hydroponically grown and intrinsically labeled with ^4SCa. Raw and cooked legumes were fed to six groups of 6-wk old male Sprague-Dawley rats. Another group received an intraperitoneal (IP) injection of ⁴⁵Ca. An eighth group was fed a casein metal extrinsically labeled with ⁴⁵Ca. The absorption of calcium from legumes by rats averaged 47.1 ± 7.5% of IP dose. Average phytate content of the legumes was 1.7% and oxalate 0.37%. Raw legumes had an average of 15000 TIA units/g of whole bean, which were completely removed by cooking. Calcium absorption was unaffected by TIA or tannin content. Reduced availability of bean calcium was likely due to phytate and/or oxalate present in legumes.     

香草醛盐酸法定量薯莨色素提取液中的缩合单宁

莨纱绸是我国华南地区特有的用一种纯手工染色和整理的,具有特殊风格和功能的丝绸面料。制作莨纱绸所使用的天然染料—薯莨色素提取物的上染量目前仍依靠人的经验控制,是影响莨纱绸品质及今后工业化生产的不利因素。本实验探究了以儿茶素为标准品,采用香草醛—盐酸法定量薯莨色素提取物中单宁的条件,结果表明:反应温度25℃,反应时间30min时,薯莨色素缩合单宁质量浓度在0.02~1.5mg/mL内与吸光值呈良好的线性关系;通过此方法制作的标准曲线计算出薯莨色素提取物中缩合单宁的含量为47.7%;该方法有较好的重复性和准确性,是一种简便、快速、可准确测定薯莨提取液中缩合单宁含量的可靠方法。     

涩柿树皮单宁的纯化及抗氧化活性

选用超声波法从涩柿树皮中提取单宁,溶剂法、大孔树脂法纯化粗提物,并考察了不同纯度的单宁对1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基和羟自由基（·OH）的清除作用。结果表明：涩柿树皮单宁的超声波提取率为5.6%;大孔吸附树脂纯化涩柿树皮单宁的最佳工艺参数为：以HPD-500树脂为吸附树脂,上柱流速1.5 mL/min,上样质量浓度1.2 mg/mL;洗脱流速1.5 mL/min,乙醇体积分数60%;粗提物、乙酸乙酯萃取物、大孔树脂纯化物、乙酸乙酯萃余物经大孔树脂纯化后产物、乙酸乙酯萃取物经大孔树脂纯化后产物的纯度分别为5.6%、11.5%、19.4%、47.4%、53.3%;当质量浓度为1 mg/mL时各纯度单宁对DPPH自由基有最大清除率,清除率分别为69.07%、80.19%、92.96%、94.02%、94.05%;清除DPPH自由基的IC50值分别为0.68、0.52、0.13、0.12、0.11 mg/mL;当质量浓度为0.9 mg/mL时各纯度单宁对·OH有最大清除率,清除率分别为43.04%、73.99%、83.00%、94.68%、96.23%。     

不同加工处理对卡亚中单宁含量影响

通过比色法研究不同加工处理对卡亚单宁含量影响,其中沸水焯烫处理效果最佳,处理后卡亚成熟叶、嫩叶单宁含量分别降至0.15%、0.05%,基本符合一般果蔬可食部分的单宁含量。     

Determination of extractable and bound condensed tannin concentrations in forage plants,protein concentrate meals and cereal grains

A procedure was developed for extraction of ‘free’ condensed tannins (CT) using a mixture of acetone/water/diethyl ether (4.7:2.0:3.3), followed by extraction of protein-bound and fibre-bound CT using boiling sodium dodecyl sulphate containing 2-mercaptoethanol (SDS). CT concentrations in all three fractions were determined by a modified butanol-HCI procedure. Separate standard curves using purified CT in water or SDS solution were utilised for analysis of extractable CT (water standards) and protein-bound and fibre-bound CT (SDS standards). The method accurately predicted the concentration of CT added to forage extracts. CT extractable in acetone/water/diethyl ether comprised, on average. 68% of total CT in a range of freeze dried forage legume samples, with most of the remainder being bound to protein. When total CT concentration was low (0.6-3.0% DM), a lower proportion was extractable (33-35%). In protein concentrate meals containing CT, the extractable, protein-bound and fibre-bound components comprised 15, 60 and 25% respectively of total CT. Total CT concentration in the forages Lotus corniculatus and Coronilla varia was considered appropriate for ruminant nutrition (2.1 and 3.0% DM). whilst CT concentration in the forage of Dorycnium spp (13–19% DM) was more suitable for soil conservation purposes. The substantial CT concentration in cottonseed meal (1.6% DM) may be involved in the high resistance of proteins in this product to ruminal degradation. CT concentration was indistinguishable from zero in perennial ryegrass forage, in barley and triticale grains and in soya bean meal (0.1% DM).     

The DMACA–HCl Protocol and the Threshold Proanthocyanidin Content for Bloat Safety in Forage Legumes

A protocol using HCl-acidified 4-dimethylaminocinnamaldehyde (DMACA) was developed for screening proanthocyanidins (PA) in plants. Individual PA-containing cells could be observed following staining of decolourised leaves with DMACA. The quantitative assay using DMACA-HCl to detect soluble PA and butanol–HCl to detect insoluble PA could detect PA content lower than 0·4 mg g⁻¹ dry matter (DM). The DMACA–HCl protocol is recommended for PA detection in low-PA plants because of its higher sensitivity than vanillin–acid protocols. Twenty-two forage legume species, most of which had been reported free of leaf PA but bloat-safe, were re-examined using the DMACA–HCl protocol. PA-positive cells were observed in leaves of 15 species. Measurable amounts of PA were detected in leaves of seven species, with the PA content ranging from 0·6 to 11 mg g⁻¹ DM. White clover ( Trifolium repens L), red clover ( T pratense L) and lucerne ( Medicago sativa L) were also found to have PA-positive leaf trichomes. By analysis of these data and the literature, the threshold PA content for bloat safety in forage legumes was estimated to be 1–5 mg PA g⁻¹ DM.     

黑木相思树皮单宁含量的分析研究及其栲胶的试制

对广西种植的4年生黑木相思树皮的单宁含量进行分析研究和黑木相思栲胶的试制.结果显示,黑木相思树皮单宁属于缩合类单宁,总固物为38.2％,单宁含量为26.0％(纯度为71.6％),其单宁含量与马占相思树皮相当,略低于黑荆树皮;生产1 t栲胶的树皮消耗量约为2.5 t,栲胶单宁含量达68.1％,纯度为70.5％.黑木相思树...     

Accounting for co‐extractable compounds (blank correction) in spectrophotometric measurement of extractable and total‐bound proanthocyanidin in Leucaena spp

Methods to account for the spectral interference of co‐extractable compounds (blank correction) in the spectrophotometric analysis of both extractable and bound proanthocyanidin (PA) using the proanthocyanidin (butanol/HCl) assay were evaluated. Crude extractable and bound PA sample matrices of PA‐free Leucaena magnifica were used. Extractable PA blanks generated in heated 95% butanol/5% H₂O reagent underestimated the optical density (absorbance) of co‐extractable compounds by 24% (P < 0.01), whereas unheated 95% butanol/5% HCl blanks, incubated at room temperature, accurately measured the absorbance of the background matrix (P < 0.01). Current procedures that estimate bound PA concentrations using the proanthocyanidin assay produce intensely coloured background matrices. Recovery measurements from total‐bound PA extracts spiked with 1071 and 2142 µg anthocyanidin per tube indicated that existing analytical procedures that do not account for the spectral interference of co‐extractable compounds overestimated (P < 0.01) bound PA concentrations by 69 and 38% respectively. An innovative technique that generated an internal correction factor for each sample, using wavelength‐scanning spectrophotometry and non‐linear curve‐fitting computer software, was developed. This procedure recovered 100% of added anthocyanidins from bound PA matrices. © 2002 Society of Chemical Industry     

核桃青皮中单宁的类型及含量测定

通过定性试验得出核桃青皮中单宁类型为水解类。以Zn2＋络合法测定普通核桃青皮和美国黑核桃青皮中单宁含量分别为19.24%和29.08%,经试验对比发现,不同干燥方式（阴干、晒干、60℃烘干）对核桃青皮中单宁含量影响较大。     

分光光度法测定板栗刺壳、核桃青皮及葡萄皮中的单宁含量

确定了邻菲罗啉分光光度法测定单宁酸含量的最佳条件,测定结果表明板栗刺壳、核桃青皮及葡萄皮中的单宁酸含量分别为6.55%、4.06%和2.10%。希望能够为这些废弃物的回收利用提供数据支持和理论依据。     

不同皮渣浸渍发酵时间对葡萄酒中缩合单宁构成及感官品质的影响

为明确不同皮渣浸渍时间对葡萄酒中缩合单宁的影响,以赤霞珠和西拉为材料,进行不同浸渍发酵时间处理,对葡萄皮和葡萄籽以及葡萄酒中缩合单宁的含量、组成、缩合单宁以及相关口感指标等进行了分析.结果 表明,不同葡萄品种中缩合单宁的初始含量、聚合度及没食子酰化率都有显著差异(P＜0.05).葡萄皮中缩合单宁的平均聚合度显著高于葡萄...     

东北不同杂豆油脂含量及脂肪酸组成分析

为开发与利用东北杂豆资源,本文以东北特有杂豆为原料,采用气质分析方法对杂豆脂肪含量及脂肪酸组成进行初步分析。结果表明：杂豆中含有丰富的不饱和脂肪酸,且在测定所有杂豆中均是高活性的n:3、n:2类不饱和脂肪酸含量高,而活性相对的n:1类不饱和脂肪酸含量低。芸豆类杂豆的脂肪酸中不饱和脂肪酸含量均大于80%;且高活性的n:3类不饱和脂肪酸含量较多,紫花芸豆的n:3类不饱和脂肪酸含量达到了48.99%,具有进一步开发应用的价值。本研究为东北杂豆的综合利用提供了理论参考与依据     

不同单宁含量的杨梅栲胶鞣革的渗透速度

采用单宁含量分别为69．4％、67，4％、65．0%、63．5％、60．5％的杨梅栲胶进行鞣革对比试验．结果表明：渗透速度与栲胶的单宁含量不成正比关系。单宁含量63．0％ 以上的栲胶，渗透速度约为0．4mm／d，与单宁含量无关；而单宁含量为60．0％的栲胶，渗透速度明显减慢，约为0．2mm／d。由于实际生产的栲胶产品单宁含量几乎都在65．0％以上，小于63．0％的极少。因此认为，市场销售的栲胶均可单独适用于鞣制重革，不会影响鞣期。     

Forage quality and tannin concentration and composition of a collection of the tropical shrub legume Flemingia macrophylla

A collection of 23 Flemingia macrophylla (Willd.) Merrill accessions of different growth habits and contrasting digestibility and one Flemingia stricta reference accession were assessed for forage quality with particular emphasis on digestibility, condensed tannin concentration and fiber content. Large differences in in vitro dry matter digestibility (IVDMD) (356 to 598 g kg^?1), content of crude protein (CP) (121 to 254 g kg^?1) and extractable condensed tannins (CT) (0 to 268 g kg^?1), protein‐binding capacity of extractable CT or astringency (1.7 to 7.9 protein‐binding entities) (PBE) and monomer composition of extractable CT were detected. IVDMD and extractable CT were negatively correlated and extractable CT was positively correlated with protein‐binding capacity. Prodelphinidin was positively and propelargonidin negatively correlated with protein‐binding capacity of extractable CT. The accessions CIAT 18438, CIAT 21083, CIAT 21090 and CIAT 22082 were superior to the most widely used accession CIAT 17403 in terms of forage quality and could be an option in production systems with acid infertile soils. In future evaluations, particular attention needs to be paid to chemical and structural features related to the composition of extractable CT and their effect on nitrogen utilization by ruminants. Copyright © 2006 Society of Chemical Industry     

Characterization of Water Extractable Crude Polysaccharides from Cherry,Raspberry, and Ginseng Berry Fruits: Chemical Composition and Bioactivity

Water extractable crude polysaccharides from three sweet cherry varieties, raspberries, and ginseng berry pulp were evaluated for their chemical composition, structural features, molecular weight, and bioactive properties. The yields of the crude polysaccharides isolated from cherries, raspberries, and ginseng berry pulp ranged from 0.79 to 0.18% on a dry weight fruit basis. All of the crude polysaccharides contained protein, phenolic compounds, and uronic acid. Each crude polysaccharide contained the sugar monomers: rhamnose, arabinose, xylose, mannose, galactose, and glucose. Of the water extractable polysaccharides obtained from the mature fruits, the crude polysaccharides from the raspberries had the highest molecular weight while the crude polysaccharides from the ginseng berry pulp had the smallest molecular weight. Results from Fourier transform infrared spectroscopy suggested that the crude polysaccharides contained a monosaccharide with six carbon atoms in a D-glucopyranose ring and a protein component. The crude polysaccharides from cherries, raspberries, and ginseng berry pulp were shown to possess antioxidant activity as determined with the ferric reducing antioxidant power and the 2,2-azinobis (3-ethyl-benzothiazoline-6-sulfonic acid) (ABTS) radical scavenging assays. The effect of crude polysaccharides on: (1) caspase 3 activation, which was determined using a hypoxia/reoxygenation model, and (2) immunostimulation, which was determined by evaluating the inflammatory mediator response, were examined. Only crude polysaccharides obtained from certain varieties of sweet cherries inhibited caspase 3 activation in a dose-dependent manner, while only the crude polysaccharides obtained from ginseng berry pulp stimulated immune function. Crude polysaccharides present in small fruits do possess bioactivities that may enhance human health.     

Effect of Heat Treatment Upon the Chemical Composition of Cottonseed Meal and Upon the Reactivity of Cottonseed Condensed Tannins

The effects of heat treatment on the chemical composition of cottonseed meal (CSM), with or without the addition of cottonseed hulls (containing condensed tannins; CT), and upon reactivity of the CT were studied. Heat was applied in a forced draught oven at 100°C for 2 h. Fluorodinitrobenzene (FDNB)-available lysine, free gossypol, extractable- and bound-CT concentrations, in vitro total nitrogen (N) solubility and the in vitro rumen degradation of the two major seed proteins (52 and 48 kDa) present in cottonseed kernel (which does not contain CT) were determined. The reactivity of CT was assessed by determining N solubility and rumen degradation of cottonseed kernel proteins in the presence or absence of polyethylene glycol (PEG; molecular weight (MW) 3500), which binds and inactivates CT. Heat treatment reduced the concentrations of free gossypol and FDNB-available lysine by small amounts, reduced measurable total CT content by 13%, reduced the solubility of total N, and reduced potential degradability of the 52 and 48 kDa cottonseed storage proteins by mixed rumen microorganisms. Addition of hulls further depressed solubility of total N and ruminal degradation of the two major storage proteins in cottonseed kernel. The action of PEG in vitro indicated that only part of the depression caused by hull addition could be explained by the presence of CT in the hulls, and that the effects of CT upon N solubility and potential degradability in heated CSM were similar to that in unheated CSM. Addition of hulls also substantially reduced FDNB-available lysine. In commercially produced materials, CSM from the Brisbane mill had a lower total CT content, lower N solubility and lower ruminal protein degradation rate than CSM from the Narrabri mill, but a similar level of FDNB-available lysine. Although application of heat inactivated 13% of the total CT, such that it could no longer be extracted and detected with butanol/HCl, it did not seem to change the overall effects produced by CT in reducing N solubility and protein degradation. The effect of hull addition in reducing available lysine has considerable relevance for feeding CSM to monogastric livestock. Interactions involving heat, hulls and CT need to be further studied.     

大孔吸附树脂分离纯化五倍子鞣质

以五倍子鞣质粗提物为原料,通过静态吸附与解吸实验比较NKA-2、NKA-9、HPD100、AB-8、D101、D301、聚酰胺及732八种吸附树脂对五倍子鞣质的吸附与解吸性能,筛选得到最优树脂NKA-2。然后通过动态吸附与解吸的单因素试验和正交试验,优化筛选NKA-2大孔吸附树脂分离纯化五倍子鞣质的工艺技术参数。结果表明,当主要考虑鞣质得率时,最优工艺参数为上样质量浓度5mg/mL、上样流速1BV/h、上样体积6BV、洗脱剂乙醇体积分数80%、洗脱流速1BV/h、洗脱体积4BV,其鞣质得率可达85.37%,纯度可达62.99%;当主要考虑鞣质纯度时,最优参数为上样质量浓度6mg/mL、上样流速2BV/h、上样体积5BV、洗脱剂乙醇体积分数80%、洗脱流速3BV/h、洗脱体积4BV,其鞣质纯度可达76.97%,得率达67.78%。