山西吕梁地区牛、羊乳及其制品的蛋白质组成分析研究

利用考马斯亮蓝G-250法定量测定了吕梁地区的牛羊乳中的蛋白质含量;采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法(SDS-PAGE)分析了牛羊乳中的蛋白质种类。结果表明:羊乳中的蛋白质含量显著高于牛乳,新鲜牛乳的碱性磷酸酶(ALP)活力和过氧化物酶(PLO)活力均显著高于新鲜羊乳。牛羊乳中蛋白质的种类差别比较的最佳试验方案:分离胶、浓缩胶浓度分别为12.5%和5%,电压分别为150 V和120 V。从电泳图谱可知:牛乳中的α-酪蛋白含量比羊乳高,而分子量却小于羊乳;牛乳中的β-酪蛋白分子量小于羊乳。因此,α-酪蛋白和β-酪蛋白可作为电泳法区别牛羊乳的特征性蛋白条带。     

不同来源乳中α-乳白蛋白、β-乳球蛋白和乳铁蛋白含量分析

本研究通过高效液相色谱-质谱联用法(HPLC-MS)对不同种乳中α-乳白蛋白、β-乳球蛋白和乳铁蛋白的含量进行测定和比较。结果显示,不同来源乳中α-乳白蛋白、β-乳球蛋白、乳铁蛋白和总蛋白存在一定差异。羊乳中乳清蛋白平均值为0.385 mg/100 g,低于牛乳和牦牛乳;羊乳中乳铁蛋白含量最高,均值为4.43 mg/100 g,最大值9.90 mg/100 g,是优质的乳铁蛋白来源;牦牛乳总蛋白含量(均值3.61%),明显高于牛乳(均值3.22%)和羊乳(均值2.89%);牛奶中乳清蛋白量在三类乳中最高。     

基于β-胡萝卜素检测的牛羊乳混掺鉴别技术

在羊乳中掺入牛乳也是乳品行业的一种掺假行为。由于牛乳中β-胡萝卜素含量远高于羊乳,本文建立了一种以β-胡萝卜素的含量作为特征指标用高效液相色谱法(HPLC)检测牛羊乳混掺的定量分析方法。乳样品皂化后,经石油醚提取、水洗浓缩后,使用岛津Shim-pack VP-ODS C18(4.6 mm×150 mm,5μm)色谱柱,以甲醇:乙腈:三氯甲烷(12:6:82,V/V/V)作为流动相洗脱,检测波长为450 nm。利用被检样纯牛羊乳中β-胡萝卜素含量变化范围,构建混掺比例线性曲线,实现定量分析。结果表明该方法的样品加标回收率为89.46%~98.19%,相对标准偏差为1.50%~2.79%。在主产季4~9月之间,牛、羊乳中β-胡萝卜素含量范围分别为0.08~0.13μg/g和1.9×10-3~2.2×10-3μg/g,线性相关系数在0.9958~0.9988范围内;盲样验证的相对误差在2.20%~4.75%之间。因此,以β-胡萝卜素为特征变量评价牛羊乳混掺比例的方法具有可行性。     

基于UPLC-QTOF-MS识别牛乳与羊乳差异

试验以小分子化学物质(分子量MW500 amu)为研究对象,基于超高效液相色谱-四级杆-飞行时间质谱仪(UPLC-QTOF-MS)建立一种识别牛乳和羊乳的检测方法。分别以甲醇和乙腈为蛋白质沉淀剂处理试样,分析正离子模式下的TIC、BPI谱图,建立主成分分析(PCA)模式,用正交偏最小二乘法判别分析(OPLS-DA)找出差异代谢产物,以仅存在于牛乳或羊乳为原则,识别出次黄嘌呤和一种m/z 491.285 5但未进一步鉴定的化合物,另外识别出2-甲基丁酰卡尼汀,在羊乳中含量高,可作为特征物质辅助鉴定牛乳与羊乳。该方法简单、便捷且灵敏度较高,为牛乳和羊乳的识别提供参考,为乳制品市场的进一步规范提供技术支撑。     

基于脂质组学法对母乳、牛乳及羊乳脂质的差异分析

基于超高效液相色谱-四极杆-飞行时间质谱技术对母乳、牛乳及羊乳的全脂质组分进行定量分析。通过分析共检出17种磷脂酰胆碱(phosphatidylcholines,PC)、11种神经酰胺(ceramide,Cer)、15种鞘磷脂(sphingomyelin,SM)、3种己糖苷神经酰胺(hexosylceramides,HexCer)、32种甘油二脂(diglyceride,DG)和2 5种甘油三酯(triglyceride, TG)。母乳与牛乳有3 6种存在显著差异的脂质,与羊乳有1 4种(P0.05)。结果表明,母乳极性脂含量(1 462.99μg/mL)显著低于羊乳而高于牛乳(P0.05),且母乳及牛乳遵循SMCerPCHexCer分布,羊乳则是CerSMHexCerPC。母乳与牛乳间存在显著差异的极性脂质主要为SM,其中SM(d14∶0/20∶0)和SM(d15∶0/24∶1)在母乳(15.90μg/mL和16.55μg/mL)中显著高于牛乳(P0.05),而羊乳与母乳间除PC(26∶0/0∶0)外,不存在其他显著差异的极性脂质。中性脂质方面,母乳中性脂质量浓度(47 749.40μg/mL)显著高于牛乳及羊乳(P0.05)。母乳与牛羊乳间存在显著性差异的中性脂质主要为TG,尤其母乳在TG(14∶0/16∶0/18∶0)与TG(16∶0/17∶0/18∶0)含量上显著低于牛羊乳(P0.05),同时3种乳在TG(17∶2/18∶0/20∶5)含量上均存在显著性差异(P0.05)。本研究不仅为婴幼儿配方奶粉的脂质研究提供理论依据,同时筛选出的差异脂质可作为鉴别牛羊乳的脂质标记物。     

牛羊乳酪蛋白制备及电泳分析比较

采采用新鲜和复原的牛羊乳为原料,等电点沉淀,通过洗涤、干燥等步骤分别制得牛乳和羊乳酪蛋白,用凯氏定氮法对其进行定量检测,应用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS—PAGE）分析比较牛羊乳酪蛋白组分差异。结果表明：牛乳酪蛋白得率为86．75％,羊乳酪蛋白得率为90．55％,高含量的酪蛋白主要集中在电泳图谱的中分子量组,可分为ds,CN、as2-CN、β-CN和K—CN,牛羊乳as2-CN分子量羊乳大于牛乳,牛乳a-CN含量比较多,羊乳β—CN含量比较多,鲜乳与复原乳全蛋白主要组分在电泳图谱中除酪蛋白差别外,上端的高分子量组清蛋白区羊乳IgG重链的分子量比牛乳小。应用蛋白质电泳分析技术可以区分羊乳和牛乳的蛋白质组分,等电点沉淀法制备酪蛋白的方法简单,易于操作。     

牛羊乳蛋白质粒度对其热处理沉淀率的影响

研究牛羊乳蛋白质粒度大小对其热处理沉淀率的影响。采用离心沉淀、原子力显微镜、激光粒度仪和SDS-PAGE分析热处理后(95℃,15 min)牛羊乳蛋白质沉淀率、表面形貌、牛羊鲜乳蛋白质粒度和酪蛋白组成。结果表明:羊乳热处理后沉淀率显著高于牛乳(p0.05);羊乳蛋白质粒子明显大于牛乳,其蛋白质热处理片状凝聚程度(35 858.15 nm~2)大于牛乳(18 215.18 nm~2);鲜牛乳蛋白质粒度大小主要集中在1 nm以下,鲜羊乳蛋白质粒度在1 nm以下相对较少,主要分布在1~1 000 nm之间;牛羊鲜乳酪蛋白主要有β-酪蛋白和αs1-酪蛋白两种,相对分子质量分别为34 000和26 000左右。牛乳的热稳定性高于羊乳;蛋白质粒度大小是影响其热稳定性的直接因素;原子力显微镜结合激光粒度仪是分析乳蛋白粒度的有效手段。     

母乳、牛乳及山羊乳脂肪酸组成的差异分析

采用气质联用法(GC-MS)测定东北区域不同泌乳期的母乳及牛乳和山羊乳常乳脂肪酸组成,并对其组成和含量进行差异分析,旨在为母乳脂质组学及以牛羊乳为基质的婴儿配方食品提供一定的理论基础。结果表明:母乳中主要脂肪酸为棕榈酸、油酸、亚油酸等,其中油酸含量最为丰富。饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)和多不饱和脂肪酸(PUFA)含量在不同泌乳期存在差异,其中SFA差异显著(p<0.05),初乳、过渡乳和成熟乳SFA含量分别为36.16%、37.89%、38.10%。牛乳和山羊乳主要以SFA为主,山羊乳SFA含量最高(69.07%),SFA中辛酸和癸酸是羊乳的特征脂肪酸,其含量显著高于母乳和牛乳(p<0.05)。山羊乳中中链脂肪酸(MCFA)含量最高(21.03%),是牛乳的1.5倍。其中,母乳SFA:MUFA:PUFA的比例为1.41:1.29:1,牛乳为19.12:9.98:1,山羊乳为11.14:3.98:1,山羊乳脂肪酸组成在比例上更加接近母乳。山羊乳在婴儿配方食品开发方面有更高的优势和开发空间。     

基于顶空气相色谱-离子迁移谱法分析脱脂处理对牛乳挥发性风味物质的影响

目的 从挥发性风味物质的角度探究影响脱脂处理后牛乳风味变化的原因。方法 采用顶空气相色谱-离子迁移谱法(headspace-gas chromatography-ion mobility spectrometry, HS-GC-IMS)构建全脂牛乳和脱脂牛乳挥发性风味物质指纹图谱, 结合主成分分析(principal component analysis, PCA)对其中挥发性风味物质差异进行分析。结果 全脂牛乳和脱脂牛乳中共检测出50个信号峰, 鉴定出26种挥发性风味物质, 主要包括6种醛、10种酮、4种醇、5种酯和1种萜烯物质。牛乳在脱脂处理后1-戊醇、2-庚酮、丁酸乙酯和2-甲基乙酸乙酯的含量降低, 而辛醛、壬醛、乙酸戊酯、E-2-庚烯醛、己酸乙酯和2-辛酮的含量则增加。结论 全脂牛乳和脱脂牛乳中挥发性成分有显著差别, 脱脂处理会通过改变牛乳中挥发性风味物质含量影响其风味品质。该研究可为脱脂乳产品营养成分的相关评价及研究提供理论依据, 也可为消费者选购乳产品提供科学参考。     

牛羊乳混掺检测鉴别技术研究进展

近年来,羊乳以其较好的营养特性渐成流行趋势,由于季节波动的影响,其价格远高于牛乳。在经济利益的驱动下,羊乳中掺入牛乳的现象时有发生,且呈现日趋严重的趋势,制约了羊奶产业的良性发展,迫切需要建立快速准确的牛羊乳混掺定性定量检测技术体系。本文对牛羊乳的差异及据此建立的、业已报道的相关检测技术进行了比较分析。常用检测技术主要包括色谱技术(气相色谱、气相色谱-质谱联用、高效液相色谱-质谱联用、高效液相色谱)、电泳技术(聚丙烯酰胺凝胶电泳、等电点聚焦电泳、毛细管电泳)、酶联免疫技术、聚合酶链式反应技术等,介绍了各种检测技术的原理及特点,并分析其可行性,为探索新的高效检测方法提供了思路,为牛羊乳混掺检测分析提供文献参考。     

A gas chromatography-mass spectrometry-based metabolomic approach for the characterization of goat milk compared with cow milk

In this work, the polar metabolite pool of commercial caprine milk was studied by gas chromatography-mass spectrometry and multivariate statistical data analysis. Experimental data were compared with those of cow milk and the discriminant analysis correctly classified milk. By the same means, differences due to heat treatments (UHT or pasteurization) on milk samples were also investigated. Results of the 2 discriminant analyses were combined, with the aim of finding the discriminant metabolites unique for each class and shared by 2 classes. Valine and glycine were specific to goat milk, talose and malic acid to cow milk, and hydroxyglutaric acid to pasteurized samples. Glucose and fructose were shared by cow milk and UHT-treated samples, whereas ribose was shared by pasteurized and goat milk. Other discriminant variables were not attributed to specific metabolites. Furthermore, with the aim to reduce food fraud, the issue of adulteration of caprine milk by addition of cheaper bovine milk has been also addressed. To this goal, mixtures of goat and cow milk were prepared by adding the latter in a range from 0 to 100% (vol/vol) and studied by multivariate regression analysis. The error in the level of cow milk detectable was approximately 5%. These overall results demonstrated that, through the combined approach of gas chromatography-mass spectrometry and multivariate statistical data analysis, we were able to discriminate between milk typologies on the basis of their polar metabolite profiles and to propose a new analytical method to easily discover food fraud and to protect goat milk uniqueness. The use of appropriate visualization tools improved the interpretation of multivariate model results.     

山羊奶与牛奶和人奶营养成分的比较

对山羊奶、牛奶和人奶中蛋白质、脂肪、维生素、矿物质等主要营养成分进行了比较,分析了3种奶中主要营养成分的差别。通过比较发现,山羊奶在总体营养成分方面优于牛奶,更接近人奶。但山羊奶存在铁、叶酸和维生素B12含量较低以及羊奶膻味的问题,应在营养上对这方面加以重视。     

Major proteins of the goat milk fat globule membrane

Fat is present in milk as droplets of triglycerides surrounded by a complex membrane derived from the mammary epithelial cell called milk fat globule membrane (MFGM). Although numerous studies have been published on human or bovine MFGM proteins, to date few studies exist on MFGM proteins from goat milk. The objective of this study was thus to investigate the protein composition of the goat MFGM. Milk fat globule membrane proteins from goat milk were separated by 6% and 10% sodium dodecyl sulfate-PAGE and were Coomassie or periodic acid-Schiff stained. Most of MFGM proteins [mucin-1, fatty acid synthase, xanthine oxidase, butyrophilin, lactadherin (MFG EGF-8, MFG-E8), and adipophilin] already described in cow milk were identified in goat milk using peptide mass fingerprinting. In addition, lectin staining provided a preliminary characterization of carbohydrate structures occurring on MFGM proteins from goat milk depending on α_S1-casein genotype and lactation stage. We provide here first evidence of the presence of O-glycans on fatty acid synthase and xanthine oxidase from goat milk. A prominent difference between the cow and the goat species was demonstrated for lactadherin. Indeed, whereas 2 polypeptide chains were easily identified by peptide mass fingerprinting matrix-assisted laser desorption/ionization-time of flight analysis within bovine MFGM proteins, lactadherin from goat milk consisted of a single polypeptide chain. Another striking observation was the presence of caseins associated with MFGM preparations from goat milk, whereas virtually no caseins were found in MFGM extracts from bovine milk. Taken together, these observations strongly support the existence of a singular secretion mode previously hypothesized in the goat.     

保加利亚乳杆菌在牛羊乳基质中发酵性能差异的比较

探讨了德氏保加利亚乳杆菌(L.Bulgaricus)在牛、羊乳基质中的增菌规律及产酸、产香性能,并对其差异性进行了比较。结果表明:以羊乳为基质42℃发酵8 h活菌数最高可达2.10×109mL-1,pH值降至4.16±0.07,滴定酸度(128.00±3.61)oT,丁二酮质量浓度(11.72±0.25)mg/L,乙醛质量浓度(6.09±0.22)mg/L;以牛乳为基质相同条件发酵活菌数最高可达1.25×109mL-1,pH值降至4.45±0.08,滴定酸度(114.80±1.31)oT,丁二酮质量浓度(11.56±0.11)mg/L,乙醛质量浓度(5.12±0.38)mg/L。结论:L.Bulgaricus以牛、羊乳为基质42℃发酵8 h,其产丁二酮能力差异不显著;而增菌效率、产酸及产乙醛能力差异极显著,L.Bulgaricus在羊乳基质中发酵性能优于牛乳。     

Mineral retention in three-week-old piglets fed goat and cow milk infant formulas

Goat milk and cow milk are commonly used in infant formula preparations and, as such, understanding the nutritional characteristics of infant formulas made from these milks is important. In this study, a goat milk infant formula was compared with an adapted (whey-enhanced) cow milk infant formula with respect to mineral absorption and deposition using the 3-wk-old piglet as a model for the 3-mo-old infant. Equal numbers of piglets (n = 8) were fed either the goat milk formula or the cow milk formula. The mineral composition of the prepared goat milk formula was higher than that of the prepared cow milk formula for most minerals, including calcium (75.1 vs. 56.7 mg/100 mL) but excluding iron, which was higher in the prepared cow milk formula (0.92 vs. 0.74 mg/100 mL). The amounts of calcium, phosphorus, and manganese absorbed by the piglets were significantly higher for the goat milk formula, whereas the amounts of zinc, iron, and magnesium absorbed were significantly higher for the cow milk formula. Apparent mineral absorption, relative to intake, was statistically higher in the cow milk formula for calcium and phosphorus, although the actual differences were very small (less than 1.3%). For copper, zinc, iron, and magnesium there was no significant difference between treatments in apparent mineral absorption, whereas for manganese, absorption was higher for the goat milk infant formula. The absolute mineral deposition was higher in piglets fed the goat milk formula for calcium, phosphorus, and manganese, whereas iron deposition was higher in the piglets fed cow milk formula. For all other minerals tested, there were no significant differences between treatments. The goat milk infant formula provided a pattern of mineral retention in the 3-wk-old piglet very similar to that of the adapted cow milk infant formula. The minor differences observed between the 2 appeared to be due to the different mineral contents of the 2 formulas.     

掺假羊乳及其制品中牛乳的检测技术研究进展

羊乳具有营养价值高、蛋白质组成更接近人乳、脂肪球直径小及致敏性低等优点, 更利于人体消化吸收, 受到消费者和乳品企业的青睐。近年来我国羊乳产业发展迅速且潜力巨大, 但由于受羊乳产量和养殖规模的限制, 羊乳价格昂贵, 市场中存在羊乳及其制品掺假牛乳的现象, 且掺假手段多样, 难以辨别。为了保证消费者的健康和权益, 保障羊乳市场良性发展, 羊乳及其制品的纯正性、真实性检测已经成为热点研究方向。本文通过分析基于乳中蛋白质、脂肪和核酸差异的羊乳中牛乳掺假检测技术的研究现状, 介绍和探讨了各检测技术的基本原理及其在应用中的优缺点, 同时展望羊乳掺假检测技术的发展方向, 旨在为牛羊乳混合掺假检测技术的进一步发展提供资料参考和思路。     

Changes in texture,composition and sensory characteristics of Camembert cheese made from a mixture of goat milk and cow milk during ripening

The study was conducted to evaluate the nutritional, textural and sensory characteristics of Camembert cheese made from goat milk, cow milk and their blends. The type of milk used affected yield, physicochemical properties and fatty acid profile of the cheeses. The percentages of C6:0, C8:0 and C10:0 acids were higher in the cheeses containing ≥ 75% goat milk. The sensory evaluation results showed differences in texture, colour, aroma and flavour among the cheeses. Cheeses made with the mixture of cow and goat milk particularly 50% goat milk mixed with 50% cow milk (50G:50C) maintained particular positive nutritional characteristics especially with respect to the mineral contents, fatty acids and consumer acceptability.     

Prediction of fatty acid profiles in cow,ewe, and goat milk by mid-infrared spectrometry

Mid-infrared (MIR) spectrometry was used to estimate the fatty acid (FA) composition in cow, ewe, and goat milk. The objectives were to compare different statistical approaches with wavelength selection to predict the milk FA composition from MIR spectra, and to develop equations for FA in cow, goat, and ewe milk. In total, a set of 349 cow milk samples, 200 ewe milk samples, and 332 goat milk samples were both analyzed by MIR and by gas chromatography, the reference method. A broad FA variability was ensured by using milk from different breeds and feeding systems. The methods studied were partial least squares regression (PLS), first-derivative pretreatment + PLS, genetic algorithm + PLS, wavelets + PLS, least absolute shrinkage and selection operator method (LASSO), and elastic net. The best results were obtained with PLS, genetic algorithm + PLS and first derivative + PLS. The residual standard deviation and the coefficient of determination in external validation were used to characterize the equations and to retain the best for each FA in each species. In all cases, the predictions were of better quality for FA found at medium to high concentrations (i.e., for saturated FA and some monounsaturated FA with a coefficient of determination in external validation >0.90). The conversion of the FA expressed in grams per 100 mL of milk to grams per 100 g of FA was possible with a small loss of accuracy for some FA.     

山羊乳与牛乳配方乳粉的致敏性比较

山羊乳是一类营养丰富且易于消化的乳品,因其营养成分与牛乳相近而受到广泛关注。本研究通过蛋白质潜在致敏性树状评估策略（生物信息学、消化稳定性、血清学研究、动物模型）对山羊乳配方乳粉的致敏性进行评价,并研究山羊乳配方乳粉与牛乳过敏患者血清中免疫球蛋白（immunoglobulin,Ig）E之间存在的交叉反应性。结果表明,山羊乳配方乳粉的167 种蛋白质中,β-乳球蛋白和α-乳清蛋白含量显著低于牛乳配方乳粉（P＜0.05）,山羊乳配方乳粉在模拟胃液中更易消化,且与过敏患者血清的结合能力弱于牛乳配方乳粉,动物模型结果表明山羊乳配方乳粉致敏小鼠体温下降幅度、临床过敏症状评分、血清中特异性IgE和IgG1抗体水平、血浆中组胺水平、血管渗透性及组织炎症程度等均显著低于牛乳配方乳粉组致敏小鼠（P＜0.05）,交叉反应性结果表明山羊乳配方乳粉能与牛乳过敏患者血清结合,但结合能力明显低于牛乳配方乳粉。结论：与牛乳配方乳粉相比,山羊乳配方乳粉的致敏性较弱,可作为牛乳的替代品,降低牛乳过敏性疾病的发病风险。