HPLC-UV法测定黄芪中黄酮类化合物含量

采用HPLC-UV法同时测定黄芪中毛蕊异黄酮苷、毛蕊异黄酮、芒柄花苷以及芒柄花素含量。样品经浸泡-超声处理，流动相为乙腈-0.1%甲酸水溶液，低压梯度洗脱；流速1.0 mL/min;进样量25μL;柱温30℃;检测波长254 nm。结果显示，毛蕊异黄酮苷在线性范围18.70～187.20μg/mL色谱峰峰面积与浓度线性关系良好，相关系数为0.999 7;毛蕊异黄酮在线性范围1.68～16.80μg/mL色谱峰峰面积与浓度线性关系良好，相关系数为0.999 4;芒柄花苷在线性范围7.50～75.00μg/mL色谱峰峰面积与浓度线性关系良好，相关系数为0.999 5;芒柄花素在线性范围4.90～49.20μg/mL色谱峰峰面积与浓度线性关系良好，相关系数为0.999 8。精密度、稳定性、适应性、重复性试验以及加样回收率均符合要求。该方法简便快捷，精密度高，重现性好，稳定性高，可用于黄芪药材中多成分含量的同时测定。     

基因型对红三叶异黄酮含量的影响

采用高效液相色谱法,对4个不同红三叶品种的异黄酮含量进行了分析,发现基因型对红三叶的异黄酮总含量影响显著,4个供试材料的异黄酮总含量为:新品系R-1>瑞德>岷山>多丽.基因型对红三叶各异黄酮单体含量变化影响不同,对大豆黄素含量的变化影响最大,对刺芒柄花素含量的变化影响最小.红三叶积累刺芒柄花素的能力最高,其次是鹰嘴豆芽...     

竞争ELISA测定三叶草中芒柄花素

三叶草是饲养家畜的优质牧草,但含有高水平植物激素——芒柄花素(7- 羟基- 4’-甲氧基异黄酮)。芒柄花素能干扰反刍动物正常繁殖功能,能导致羊的繁殖机能失调,引起流产。目前一般采用高效液相色谱法、气相色谱法及H~3标记放射免疫测定法来测定三叶草中芒柄花素。这些方法尽管检测灵敏度较高,但仪器昂贵、操作环境要求严格,难以推广。而用酶联免疫吸附测定(Enzyme- linked     

红三叶与白三叶春季植物雌激素含量和种类的差异

用2个红三叶品种和3个白三叶品种检测了春季茎叶中主要植物雌激素的种类及其含量的变化。结果表明,春季红三叶茎叶植物雌激素总含量为3.35×103~7.99×103μg/g,远高于白三叶的0.57×102~3.81×102μg/g。红三叶茎叶主要含有芒柄花素、美皂异黄酮、染料木素和大豆黄酮等植物雌激素,而白三叶茎叶则主要含有芒柄花素和染料木素,时有少量美皂异黄酮,未检测到大豆黄酮。两种三叶草春季植物雌激素总含量的变化均取决于芒柄花素的变化,与生育进程无明显关系。     

紫花苜蓿中主要黄酮类化合物抗氧化活性研究

为了对紫花苜蓿中槲皮素、木犀草素、染料木素、芒柄花黄素等黄酮类成分的抗氧化能力进行综合评价,试验分别测定了紫花苜蓿中不同黄酮成分的还原能力、DPPH自由基清除能力以及铁还原抗氧化能力(FRAP)。结果表明:苜蓿中的槲皮素和木犀草素抗氧化能力最强,而染料木素的抗氧化能力在黄酮类化合物中相对较弱,芒柄花黄素的抗氧化能力最弱。     

基于主成分分析的膨化大豆品质评价

为对膨化大豆的品质进行分析及量化评价,文章以73个膨化大豆为试验材料,测定分析其蛋白质溶解度、脲酶活性、胰蛋白酶抑制因子、β-伴大豆球蛋白、大豆球蛋白、棉籽糖、水苏糖、大豆苷、黄豆黄苷、染料木苷、异黄酮总含量、大豆皂苷等含量,并运用主成分分析法简化分析评价指标,构架评价体系,得到每种样品的综合得分。结果表明:膨化大豆蛋白溶解度、脲酶活性、胰蛋白酶抑制因子、β-伴大豆球蛋白、大豆球蛋白、棉籽糖、水苏糖、大豆苷、黄豆黄苷、染料木苷、大豆异黄酮和大豆皂苷的含量范围为67.93%~87.46%,0.00~0.01 U/g,5.40~32.60 mg/g,3.10~79.50 mg/g,20.30~93.50 mg/g,6.87~21.36 mg/g,21.85~39.39 mg/g,586.96~1 101.13μg/g,66.56~205.57μg/g,409.33~750.70μg/g,1.15~1.99 mg/g,9.34~15.63 mg/g。同时,评价指标之间存在不同程度的相关性,其中脲酶活性与大豆苷、染料木苷、大豆异黄酮呈显著正相关,与黄豆黄苷显著负相关;胰蛋白酶抑制因子与大豆皂苷呈显著正相关,与大豆苷、染料木苷、大豆异黄酮呈显著负相关;大豆球蛋白与黄豆黄苷、大豆皂苷呈显著正相关,与染料木苷呈显著负相关;水苏糖与染料木苷呈显著负相关;黄豆黄苷与大豆皂苷呈显著负相关。通过主成分分析提取了前4个主成分,累计贡献率为69.928%,说明前4个主成分能够代表原来12个指标的信息,并得到评价公式Z=0.307Z1+0.174Z2+0.128Z3+0.090Z4,计算出膨化大豆的综合得分,并进行验证。     

黄芪发酵前后黄酮含量及指纹图谱比较研究

应用HPLC-UV法比较发酵前后黄芪中4种黄酮含量及色谱图变化,旨在为发酵黄芪制品提供质控数据。采用色谱条件为Inertsustain C18色谱柱(150 mm×4.6 mm,5μm),流动相为乙腈-水系统,梯度洗脱,检测波长:254 nm,柱温:25℃,流速:1 mL/min,进样量:20μL。该方法中毛蕊异黄酮苷在12.34~790 mg/L、刺芒柄花苷在5.0~320 mg/L、毛蕊异黄酮在4.53~290 mg/L、芒柄花素在9.53~610 mg/L与峰面积呈良好的线性关系,不同成分色谱峰分离程度较好。测定结果表明,黄芪经发酵后,毛蕊异黄酮苷、毛蕊异黄酮和芒柄花素含量发生显著变化,其中发酵黄芪含量分别为12.236±0.232、0.201±0.021和0.737±0.041 mg/g,对照黄芪含量分别为0.327±0.013、5.453±0.078和12.847±0.118 mg/g;发酵前后色谱图差异显著,说明生物发酵可使黄芪成分及其含量发生较大变化,试验可为黄芪发酵制品的质量控制提供借鉴。     

黄芪3种成分对H_2O_2诱导Chang liver细胞凋亡的抑制作用

以H2O2建立Chang liver细胞损伤模型,设空白对照组、损伤模型组、黄芪甲苷保护组、毛蕊异黄酮葡糖苷保护组、芒柄花素保护组和阳性对照组。采用四甲基偶氮唑盐(MTT)法、微板法、流式细胞术、DNA Ladder检测等方法测定细胞存活率、细胞外液转氨酶活性、周期分布及凋亡情况。结果显示H2O2的诱导使Chang liver细胞存活率降低、活性升高、发生G0/G1期阻滞及细胞外液转氨酶、细胞凋亡率的升高,3种成分对细胞的预处理均可显著或极显著的升高细胞存活率、缓解G0/G1期的阻滞现象、并降低细胞外液转氨酶活性及细胞凋亡率(P0.05或P0.01)。表明黄芪主要成分(黄芪甲苷、毛蕊异黄酮葡糖苷和芒柄花素)对H2O2诱导的Chang liver细胞凋亡均有一定的抑制作用。     

异黄酮植物雌激素对雌性动物生殖、泌乳和产蛋的影响及其作用机制探讨

异黄酮植物雌激素是主要存在于豆科牧草和大豆的异黄酮类化合物,有雌激素样活性(约为17β-E2的10-3~10-5),对抗癌、抗氧化,改善心血管机能等有良好的作用。对雌性动物的生殖、泌乳和产蛋的效应,视使用剂量、持续时间和动物所处的生理状态,而呈现正负双向作用。异黄酮植物雌激素(本文涉及芒柄花素、大豆黄酮和染料木素)可与机体的E2受体竞争性结合,通过神经内分泌系统的下丘脑-垂体-性腺轴调控生殖、泌乳和产蛋机能,并在生长轴、甲状腺轴等参与下,促使相关代谢激素参与其营养过程。异黄酮植物雌激素作为生理调节剂,在饲料工业中有良好的应用前景,但必须谨慎,应通过进一步的生产试验和观察。     

大孔树脂对苜蓿中4种黄酮类化合物的吸附特性

笔者以4种黄酮类化合物为评价指标.采用高效液相色谱检测试验中各个黄酮类化合物含量的变化,考察10种型号大孔吸附树脂对苜蓿中4种黄酮类化合物(槲皮素、木犀草素、染料木素、芒柄花黄素)的吸附特性,从中筛选出吸附效果最好的树脂。结果表明.HPD-722和HPD-300这2种树脂对苜蓿中黄酮的吸附效果良好。动力学试验中,HPD-722效果最好。     

红车轴草异黄酮对肉鸡生产性能及肉品质的影响

为了研究红车轴草（Trifolium pratense）异黄酮对肉鸡生长、屠宰性能及肉品质的影响,将180羽1日龄AA肉仔鸡分成3组,即对照组、试验Ⅰ组、试验Ⅱ组,分别饲喂基础日粮、基础日粮+红车轴草异黄酮(10 mg/kg)、基础日粮+红车轴草异黄酮(20 mg/kg)。结果显示,红车轴草异黄酮可以提高肉鸡的日增体质量（P<0.05）和屠宰性能（P>0.05）,降低血清中甘油三酯、总胆固醇和腹脂率（P<0.05）,减少了储存第6天肉中丙二醛含量（P<0.05）,降低了肉品滴水损失和剪切力（P>0.05）。表明红车轴草异黄酮能够提高肉鸡生长性能和屠宰性能,改善肉品品质,添加20 mg/kg时肉鸡日增长速度最快。     

岷山红三叶异黄酮对去卵巢大鼠生长和免疫及抗氧化指标的影响

为研究岷山红三叶异黄酮的促生长、免疫及抗氧化作用,选择80只SD(Sprague-Dawley)雌性大鼠,随机分为8组,每组10只,其中1～7组为去卵巢+红三叶异黄酮组, 分别按320,160,80,40,20,10和0 mg/(kg BW·d)的量灌胃红三叶异黄酮,第8组为假手术组,灌服等量生理盐水。结果表明,1)红三叶异黄酮对去卵巢雌性大鼠生长有一定的抑制作用,但不存在量效关系。2)低剂量的红三叶异黄酮具有提高大鼠T淋巴细胞转化率的作用,可以作为畜禽免疫增强剂,提高畜禽免疫力,以10 mg/(kg BW·d)的添加量比较适宜。3)岷山红三叶异黄酮具有明显的抗氧化作用,适量(>80 mg/kg BW·d)添加可降低血清CHO(total cho1esterol, 总胆固醇)、HDL(high density lipoprotein cholesterol,高密度脂蛋白胆固醇)和LDL(low density lipoprotein cholesterol,低密度脂蛋白胆固醇)浓度,提高血清SOD(superoxide dismutase, 超氧化物歧化酶)含量。4)岷山红三叶异黄酮具有弱的雌激素作用和较好的使用安全性。5)综合各指标,岷山红三叶异黄酮的适宜添加量为80 mg/(kg BW·d)。     

Effects of Phytooestrogens Supplementation on Blood Biochemical Indexes and Antioxidant Capacity of Lactating Yili Mare

The experiment was conducted to study the effects of phytooestrogens supplementation on blood biochemical indexes and antioxidant capacity of Yili maria during lactation.A total of 24 Yili maria during lactation were divided into four groups with similar age,parity,lactation months,i.e.groups Ⅰ,Ⅱ,Ⅲ and control group.The control group and trial groups fed with the same based diet,groups Ⅰ,Ⅱ and Ⅲ were supplemented with 0.7 g soybean isoflavone,0.7 g genistein,0.35 g soybean isoflavone and 0.35 g genistein respectively.The total duration of the experiment was 90 d,the first 10 d of the experiment was the preliminary period and the formal experiment lasted for 80 d.The blood samples were collected by jugular vein in the experiment of 0,20,40,60,80 days,respectively,3 500 r/min centrifuged 15 min separate plasma and -20 ℃ saved for later use.The results showed that compared with control group,total protein,albumin content and GSH-Px,SOD,CAT activity of blood in trail groups were no significant difference (P >0.05);But CAT,T-AOC activity of blood in trail groups had a upward trend.GLB,BUN content of blood in group Ⅱ were significantly higher than control group and group Ⅰ (P< 0.05);no significant difference was observed among other groups (P >0.05).Compared with control group,the MDA content of blood in groups Ⅰ and Ⅱ were trends to decrease,and group Ⅰ reached significant level (P< 0.05).Therefore,phytooestrogens supplementation had enhanced immune function,improved the body's antioxidant capacity of Yili maria during lactation,when supplementary feeding 0.7 g soybean isoflavone on antioxidant capacity of Yili maria during lactation was more obvious effect.     

补饲植物雌激素对泌乳期伊犁母马血液生化指标及抗氧化力的影响

试验旨在研究补饲植物雌激素对泌乳期伊犁母马血液生化指标及抗氧化能力的影响。根据年龄、胎次、泌乳月龄相近的原则选择泌乳期伊犁母马24匹,随机分为4组,分别为对照组、试验Ⅰ组、试验Ⅱ组和试验Ⅲ组。对照组饲喂基础日粮,试验Ⅰ、Ⅱ、Ⅲ组在饲喂基础日粮的基础上分别补饲0.7 g大豆异黄酮、0.7 g染料木素、0.35 g大豆异黄酮和0.35 g染料木素。预试期10 d,正试期80 d,分别于试验的第0、20、40、60、80天颈静脉采集血液样品,3 500 r/min离心15 min,分离血浆,－20 ℃保存备用。结果表明,与对照组相比,补饲植物雌激素对泌乳期伊犁母马血液中总蛋白、白蛋白含量和谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶活性的影响均不显著(P >0.05);但试验组过氧化氢酶活性、总抗氧化能力均有不同程度的提高;试验Ⅱ组血液中球蛋白、尿素氮含量显著高于对照组和试验Ⅰ组(P<0.05),其余各组间差异均不显著(P >0.05);与对照组相比,试验Ⅰ、Ⅱ组血液中丙二醛含量均呈降低趋势,且试验Ⅰ组达到显著水平(P<0.05)。因此,补饲植物雌激素对泌乳期母马增强免疫机能,提高机体抗氧化能力起到了一定作用,且补饲0.7 g大豆异黄酮时对机体抗氧化能力效果较为明显。     

内源异黄酮对紫花苜蓿结瘤固氮及氮效率的调控研究北大核心CSCD

为探究紫花苜蓿体内异黄酮与结瘤固氮及氮效率的关系,以紫花苜蓿品种中氮高效的"LW6010"和氮低效的陇东苜蓿为材料,采用室内营养液砂培,设5个氮素水平,研究了不同氮效率紫花苜蓿的异黄酮含量和固氮特性以及异黄酮合酶(IFS)和结瘤信号传递通路基因(nod)的表达。结果表明:紫花苜蓿的异黄酮含量与根瘤数、单株总根瘤重以及单株固氮潜力的相关性显著,并且在低氮胁迫时紫花苜蓿的异黄酮含量以及根瘤数、单株总根瘤重和单株固氮潜力均显著上升。另外,还发现:氮高效的"LW6010"紫花苜蓿在氮胁迫时,茎叶和根系中上调的IFS基因数量和上调幅度均大于氮低效的陇东苜蓿,因此促进了nod基因的高表达,最终提高了氮效率。综上所述,1)紫花苜蓿的结瘤固氮与异黄酮含量显著正相关,在外源氮素浓度改变时,紫花苜蓿能通过茎叶和根系中不同IFS基因的表达提高体内异黄酮含量,从而刺激nod基因表达来促进结瘤和提高单株固氮潜力;2)紫花苜蓿体内异黄酮含量的差异是决定其氮效率高低的重要原因,氮高效紫花苜蓿在氮胁迫时通过多个IFS基因的高表达促进异黄酮合成,进而刺激结瘤固氮,从而表现出氮高效。     

Effects of dietary soy isoflavones on growth, carcass traits, and meat quality in growing-finishing pigs

Two experiments were conducted to determine the effect of soy isoflavones on growth, meat quality, and carcass traits of growing-finishing pigs. In Exp. 1, 36 barrows (initial and final BW, 26 and 113 kg, respectively) were used and each treatment was replicated four times with three pigs each. The dietary treatments were 1) corn-soybean meal (C-SBM), 2) corn-soy protein concentrate (low isoflavones, C-SPC), or 3) C-SPC + isoflavones (isoflavone levels equal to those in C-SBM). Daily gain and ADFI were increased (P < 0.10) in pigs fed the C-SPC relative to pigs fed the C-SPC + isoflavone diet in the late finishing period; otherwise, growth performance was not affected (P > 0.10) by diet. Longissimus muscle area, 10th-rib fat depth, percentage muscling (National Pork Producers Council), 24-h pH and temperature, color, firmness-wetness, marbling, drip loss, and CIE L*, a*, and b* color values were not affected (P > 0.10) by diet. Dressing percentage, carcass length, weight and percentage of fat-free lean in ham and carcass, lean gain per day, lean:fat, and ham weight were increased (P < 0.10), and ham fat and percentage fat in ham and carcass were decreased (P < 0.10) in pigs fed the C-SPC + isoflavone diet compared with pigs fed the C-SPC diet. Pigs fed the C-SPC + isoflavone diet had similar (P > 0.10) carcass traits as pigs fed the C-SBM diet, except carcass length, percentage ham lean and thaw loss were greater (P < 0.10), and total ham fat was less (P < 0.10) in pigs fed the C-SPC + isoflavone diet. In Exp. 2, 60 gilts (initial and final BW, 31 and 116 kg, respectively) were used, and each treatment was replicated five times with four pigs per replicate. The treatments were 1) C-SBM, 2) C-SBM + isoflavone levels two times those in C-SBM, and 3) C-SBM + isoflavone levels five times those in C-SBM. Daily feed intake was linearly decreased (P < 0.10) in the growing phase and increased (P < 0.10) in the late finishing phases as isoflavone levels increased; otherwise, growth performance was not affected (P > 0.10) by diet. Diet did not affect (P > 0.10) carcass traits; however, CIE a* and b* color scores and drip loss were decreased (P < 0.06) as isoflavone levels increased. Soy isoflavones decreased fat and increased lean in barrows when fed within the dietary concentrations found in typical C-SBM diets but not when fed to gilts at concentrations above those present in C-SBM diets.     

Exposure of growing and adult captive cheetahs (Acinonyx jubatus) to dietary isoflavones: twenty years later

Dietary isoflavones are associated with oestrogenic and anti‐oestrogenic effects, and have been linked to infertility in cheetahs. This study aimed to determine the isoflavone content of commercially prepared diets consumed by captive cheetahs. Sixteen international zoological facilities provided diets, and the isoflavone content of each diet was determined by acid hydrolysis and HPLC quantification. Proximate nutritional composition was also determined. Over half the diets analysed contained detectable concentrations of isoflavones, whereby total isoflavone content ranged from 1.75–183 mg/kg dry matter. The zoo‐specific diets were calculated to deliver a median isoflavone dose of 0.07 mg/kg body weight (BW) and a maximum of 1.95 mg/kg BW to captive cheetahs. On a metabolic body weight basis this equates to a maximum of 4.90–5.43 mg/kg^0.75. Some diets prepared for hand‐rearing neonatal cheetahs could expose neonates to doses of up to 4.24 mg/kg BW (or 4.24–6.33 mg/kg^0.75 for cubs under 3 months of age). Only one of six zoo‐specific diets was found to deliver isoflavones in doses shown to possess biological activity in other species. Therefore, on average, dietary isoflavones were not found in commercially prepared diets consumed by captive cheetahs in concentrations predicted to cause physiological changes. However, a small proportion of these diets, including hand‐rearing formulas, contained elevated isoflavones concentrations which may influence cheetah fertility, behaviour or other physiological parameters.     

大豆异黄酮在畜禽养殖中的应用及研究

大豆异黄酮是大豆中富含的一类黄酮类化合物。由于其具有较弱的雌激素活性．大豆异黄酮在治疗和预防与雌激素有关的疾病方面具有重要的功效。大豆异黄酮还具有促进畜禽日增重、提高饲料利用率和生产性能以及促进畜禽免疫机能等生理功能。介绍了大豆异黄酮的性质、作用机理、在畜禽养殖中的应用及在医学中的功效。     

红车轴草异黄酮对肉鸡免疫器官、免疫球蛋白及抗氧化性能的影响

通过在饲料中添加红车轴草(Trifolium pretense L.)异黄酮,研究其对AA肉鸡免疫器官、免疫球蛋白及抗氧化性能的影响。选用180羽1日龄AA肉仔鸡分成3组,即对照组、处理Ⅰ组、处理Ⅱ组,分别饲喂基础日粮、基础日粮+红车轴草异黄酮(10 mg·kg^-1)、基础日粮+红车轴草异黄酮(20 mg·kg^-1)。结果表明:处理组的肉鸡日增重和免疫器官指数均高于对照组,饲喂21 d后,处理组血清IgM的含量显著高于对照组(P<0.05),49 d后,处理Ⅰ组肉鸡血清IgG含量显著高于对照组(P<0.05);红车轴草异黄酮提高了血清中谷胱甘肽过氧化物酶(GSH-PX)活性,降低了丙二醛(MDA)含量,处理Ⅰ组和处理Ⅱ组之间差异不显著。红车轴草异黄酮能够明显改善肉鸡生产性能、调节免疫和提高机体的抗氧化性。     

红三叶个体水平异黄酮含量的差异及其形态特征和草产量比较研究

为探索选育高异黄酮含量红三叶(Trifolium pratense L.)新种质材料的可行途径，本研究测定和林格尔县实验地中300株红三叶地上部分的异黄酮含量，采用K-means聚类分析方法，将不同异黄酮含量红三叶聚类为4个类群(T1～T4),同时对4个红三叶类群进行形态特征进行观测，以及对各农艺性状和产量指标进行测定。结果表明：300株红三叶异黄酮含量介于15.65～34.16 mg·g^-1,平均含量为24.88 mg·g^-1。不同异黄酮含量的红三叶类群在形态特征、农艺性状、草产量等方面具有显著性差异，其中高异黄酮含量红三叶群体占300株的18.67%,平均含量为29.61 mg·g^-1,具有绝对株高和自然株高差值大于15 cm(平卧型)和叶片面积小于350 mm²的形态特征；同时绝对株高、草产量也显著更高(P<0.05)。本研究为红三叶高产优质栽培和品种选育奠定了基础。