SPME-GC-MS结合组学技术分析发酵椰奶特征风味与风味物质相关性

为了分析发酵椰奶特征风味与风味物质相关性,利用固相微萃取—气相色谱—质谱联用技术(Solid-Phase Micro Extraction-Gas Chromatography-Mass Spectrometry,SPME-GC-MS)分别对发酵椰奶、原味椰奶和酸牛奶中的风味成分进行测定,同时进行感官评价;并且结合组学技术从感官和风味物质方面分析了发酵椰奶的特征风味。研究结果表明,发酵椰奶感官风味特征在甜味、酸味、椰子味、奶香味、乳脂味以及醇香味感官评分较高,对发酵椰奶的喜好感主要受甜味、酸味、椰子味以及香草味的影响;而相比原味椰奶甜味过高,且伴随有较浓蒸煮味。从发酵椰奶中共检测出23种风味成分,比原味椰奶和酸牛奶分别多5种和2种;相比原味椰奶,发酵椰奶中酸类和酯类物质相对含量分别减少了11.005%和11.670%,而醇类、酮类和醛类物质相对含量分别增加了6.015%,10.295%,1.975%;酸牛奶中酸类物质是其主要风味成分,比发酵椰奶相对含量高31.79%。发酵椰奶中2,3-戊二酮、3-羟基-2-丁酮等风味成分主要贡献了果香味、奶香味和焦糖气味,酯类物质主要贡献了香草味、甜味、椰子味和蒸煮味。     

Mouthfeel and flavour of fermented whey with added hydrocolloids

High-methoxy pectin (HMP), propylene glycol alginate (PGA), carboxymethyl cellulose (CMC) and xanthan gum (XG) were added separately to whey fermented by a yoghurt starter culture at concentrations that yielded a viscosity similar to that of commercial fermented milk beverages. Mouthfeel and flavour of the samples were assessed by a highly trained panel, and volatile analysis was performed by proton-transfer reaction mass spectrometry to determine the effect of hydrocolloids on the partition of flavour compounds in the headspace. CMC and HMP produced significantly higher thickness than PGA and XG; PGA caused a ‘gritty’ mouthfeel. The yoghurt aroma of the samples containing CMC and PGA was perceived as less intense compared with the control, consistent with a reduced concentration of key volatile compounds in the headspace of these samples.     

基于气相色谱-质谱联用技术分析酸浆豆腐加工过程中风味物质的变化

目的以酸浆豆腐为研究对象,结合气相色谱-质谱联用法(gas chromatography-mass spectrometry,GC-MS)优化酸浆豆腐不同加工阶段中挥发性风味物质的捕获方法,对酸浆豆腐的加工阶段进行分类。方法采用顶空-固相微萃取法(head space-solid phase microextraction,HS-SPME)结合GC-MS对风味物质进行检测,优化风味物质捕获方法,内标法进行定量,结合聚类分析对加工阶段进行分类。结果不同批次酸浆豆腐中风味物质变化具有较高的稳定性,酸浆豆腐加工过程中共鉴定出90种挥发性风味物质,制浆、煮浆、点浆、蹲脑10 min、蹲脑20 min和压制工序分别鉴定出27、25、32、39、45和22种挥发性风味物质,主要为醛类、酮类、醇类等风味物质。对加工过程中风味物质进行聚类分析发现:适当延长煮浆与蹲脑时间,可促进更多良好风味物质的形成。结论该方法操作简单,适合酸浆豆腐中风味物质的测定且稳定性好,在后续实验中可进一步对加工工艺进行调控。     

发酵乳酪乳清中风味物质的分析研究

采用顶空固相微萃技术(HS-SPME)对发酵乳酪乳清中挥发性风味物质进行提取,并利用气相色谱-质谱联用技术(GC-MS)对挥发性风味物质进行分离鉴定。共鉴定出30种化合物,其中酸类物质质量分数最高,占总量的19.85%;其次为酮类,占总量的12.55%。在发酵乳酪乳清中这些挥发性物质中酸类和酮类物质可能为其重要的风味特征物质。     

Volatile aroma composition of distillates produced from fermented sweet and acid whey

Lactose within whey can be fermented and distilled to produce a potable distilled spirit. The aim of this study was to determine if acid and sweet whey types can be fermented and distilled using similar processes and to investigate differences in volatile aroma compounds for the 2 distillates. Fermentation and distillation of the 2 whey types progressed in a similar manner, using Kluyveromyces marxianus for the initial fermentation and a glass still fitted with a Vigreux column for the subsequent distillation. Ethanol content of the wash (fermented whey) varied considerably following each fermentation and ranged from 1.2 and 2.0% (wt/wt) with no clear trend between acid and sweet whey samples. Volatile aroma compounds were extracted using headspace solid-phase microextraction and identified via gas chromatography-mass spectrometry. Acid and sweet whey distillates contained unique volatile aromatic compounds, and significant differences in compound peak areas were observed. These differences may have an effect upon the organoleptic qualities of spirits produced from whey; therefore, whey source may be an important factor when fermenting and distilling whey.     

乳酸菌发酵核桃酸乳不良气味的形成与分析

目的：为探究核桃发酵酸乳不良气味物的形成机制及其组成,改善其风味品质。方法：采用对比实验和感官评价分析不同发酵剂、不同原料组分对发酵乳不良气味产生的影响,研究核桃乳发酵过程中原料组分、羰基价与挥发性化合物的变化规律与相关性;采用顶空-固相微萃取-气相色谱-质谱联用技术分析核桃酸乳挥发性化合物组成,采用相对气味活度值和主成分分析法对关键性风味物和不良气味物进行分析。结果：原料中脂肪组分与不良气味产生显著相关,发酵过程中酸乳羰基价与不良气味形成显著相关;核桃酸乳主要的挥发性风味物包括：己酸等3 种酸类化合物,乙偶姻等3 种酮类物,正己醇等7 种醇类化合物,(E)-2-庚烯醛等4 种醛类化合物以及丁酸丁酯和其他类化合物;相对气味活度值大于1的挥发物为(E)-2-庚烯醛、乙偶姻、己酸、正己醇、正辛醇、1-庚醇、苯甲醛、2-壬酮、2-庚酮、苯乙烯和苯乙醇;关键风味物为乙偶姻、己酸、(E)-2-庚烯醛、正己醇。结论：己酸、(E)-2-庚烯醛、正己醇、苯甲醛为核桃酸乳不良气味的主要风味物,其形成的主要路径为核桃乳发酵过程脂肪水解产生较多的亚油酸、亚麻酸,在脂肪氧合酶作用下氧化,裂解产生C6～C9的醇、醛、酸类物。     

Metabolomics of acid whey derived from Greek yogurt

Acid whey, a byproduct of Greek yogurt production, has little commercial value due to its low protein content and is also environmentally harmful when disposed of as waste. However, as a product of microbial fermentation, acid whey could be a rich source of beneficial metabolites associated with fermented foods. This study increases understanding of acid whey composition by providing a complete metabolomic profile of acid whey. Commercial and laboratory-made Greek yogurts, prepared with 3 different bacterial culture combinations, were evaluated. Samples of uncultured milk and cultured whey from each batch were analyzed. Ultra-high-performance liquid chromatography–tandem mass spectrometry metabolomics were used to separate and identify 477 metabolites. Compared with uncultured controls, acid whey from fermented yogurt showed decreases in some metabolites and increases in others, presumably due to the effects of microbial metabolism. Additional metabolites appeared in yogurt whey but not in the uncultured control. Therefore, the effect of microbial fermentation is complex, leading to increases or decreases in potentially bioactive bovine metabolites while generating new microbial compounds that may be beneficial. Metabolite production was significantly affected by combinations of culturing organisms and production location. Differences between laboratory-made and commercial samples could be caused by different starting ingredients, environmental factors, or both.     

商业发酵剂中乳酸乳球菌发酵风味特性及其化学表征的研究

为深入分析不同乳酸乳球菌在发酵乳中的风味贡献和特征香气成分,对多种商业乳酸菌发酵剂中分离得到21株乳酸乳球菌,进行单菌株发酵。根据感官评价结果进行香型分析,之后采用气相色谱-离子迁移谱仪（gas chromatograph-ion mobility spectrometry,GC-IMS）技术测定了发酵乳的挥发性风味化合物,并利用化学计量学分析方法进行风味物质的分析。结果表明:这些单菌株发酵乳可划分为奶香味、奶油味、酪香味三个香型。GC-IMS共鉴定出25种化合物,其中醇类9种、酮类8种、醛类3种、羧酸1种、酯4种。通过绘制GC-IMS指纹图谱,明确了不同菌株发酵乳的特征化合物组成。利用主成分分析（principal component analysis,PCA）,将其中4株乳酸乳球菌双乙酰变种的发酵乳进行了很好的区分,且与感官评价结果相一致。采用偏最小二乘回归分析（partial least squares regression,PLSR）对感官香味分型和挥发性成分的相关性进行分析,得到对酪香味影响比较大的挥发性风味成分有正己醛、正己醇;对奶油味贡献较大的成分有乙酸乙酯;对奶香味贡献大的成分较为分散,关键性成分有待于进一步确定。本研究为发挥乳酸乳球菌改善发酵乳风味的作用提供了借鉴和参考。     

Improved Acid, Flavor and Volatile Compound Production in a High Protein and Fiber Soymilk Yogurt-like Product

The effects of added caseinate (CAS), casein hydrolyzate (CASHY) and whey protein hydrolyzate (WPHY) on acid, flavor and volatile compound production in a high protein and fiber soymilk yogurt-like product were studied. High protein and fiber soymilk, produced by blending soaked, boiled and dehulled soybeans with Swiss cheese whey ultrafiltration permeate, was fermented with a mixed S. thermophilus and L. bulgaricus yogurt culture. The concentrations of lactic acid, key volatile compounds, i.e., acetaldehyde, acetone, and diacetyl, and the flavor and texture of the resulting soymilk based yogurt formulated with added CAS or CASHY were comparable to those in a milk yogurt control.     

Effect of Inoculation Level of Lactobacillus rhamnosus and Yogurt Cultures on Conjugated Linoleic Acid Content and Quality Attributes of Fermented Milk Products

ABSTRACT:  The effect of inoculation concentration of Lactobacillus rhamnosus and yogurt cultures and storage time on conjugated linoleic acid (CLA) content and quality attributes of fermented milk products was determined. Yogurt culture ( Lactobacillus bulgaricus and Streptococcus thermophilus, 1:1 ratio, YC ) , L. rhamnosus (LB), and LB co-cultured with yogurt culture, were inoculated at 10⁶, 10⁷, 10⁸ CFU/mL into a milk with hydrolyzed soy oil as the lipid source. CLA content, microbial counts, acidity, texture, and volatile flavor profile of the fermented milk products were stable during storage at 4 °C for 14 d. Total CLA contents ranged from 0.51 to 1.00 mg CLA/g lipid following 14 d of storage. Inoculation level of L. rhamnosus and yogurt cultures had no significant effect on CLA content and texture, but affected acidity and the volatile flavor profile of the fermented milk products. The fermented milk products produced by L. rhamnosus co-cultured with yogurt culture with 10⁷ CFU/mL total inoculation level resulted in a high CLA content and desirable quality characteristics. This research demonstrated that the optimal inoculation concentration and the combination of L. rhamnosus and yogurt cultures were important factors to produce fermented milk products with CLA content and acceptable quality attributes.     

Comparison of composition, sensory, and volatile components of thirty-four percent whey protein and milk serum protein concentrates

The objectives of this study were to identify and compare the composition, flavor, and volatile components of serum protein concentrate (SPC) and whey protein concentrate (WPC) containing about 34% protein made from the same milk to each other and to commercial 34% WPC from 6 different factories. The SPC and WPC were manufactured in triplicate with each pair of serum and traditional whey protein manufactured from the same lot of milk. At each replication, SPC and WPC were spray dried (SD) and freeze dried (FD) to determine the effect of the heat used in spray drying on sensory properties. A trained sensory panel documented the sensory profiles of rehydrated SD or FD powders. Volatile components were extracted by solid-phase microextraction (SPME) and solvent extraction followed by solvent-assisted flavor evaporation (SAFE) with gas chromatography-mass spectrometry and gas chromatography-olfactometry. Whey protein concentrates had higher fat content, calcium, and glycomacropeptide content than SPC. Color differences (Hunter L, a, b) were not evident between SPC and WPC powders, but when rehydrated, SPC solutions were clear, whereas WPC solutions were cloudy. No consistent differences were documented in sensory profiles of SD and FD SPC and WPC. The SD WPC had low but distinct buttery (diacetyl) and cardboard flavors, whereas the SD SPC did not. Sensory profiles of both rehydrated SD products were bland and lower in overall aroma and cardboard flavor compared with the commercial WPC. Twenty-nine aroma impact compounds were identified in the SPC and WPC. Lipid and protein oxidation products were present in both products. The SPC and WPC manufactured in this study had lower total volatiles and lower concentrations of many lipid oxidation compounds when compared with commercial WPC. Our results suggest that when SPC and WPC are manufactured under controlled conditions in a similar manner from the same milk using the same ultrafiltration equipment, there are few sensory differences but distinct compositional and physical property differences that may influence functionality. Furthermore, flavor (sensory and instrumental) properties of both pilot-scale manufactured protein powders were different from commercial powders suggesting the role of other influencing factors (e.g., milk supply, processing equipment, sanitation).     

Volatile flavor compounds in yogurt: a review

Considerable knowledge has been accumulated on the volatile compounds contributing to the aroma and flavor of yogurt. This review outlines the production of the major flavor compounds in yogurt fermentation and the analysis techniques, both instrumental and sensory, for quantifying the volatile compounds in yogurt. The volatile compounds that have been identified in plain yogurt are summarized, with the few key aroma compounds described in detail. Most flavor compounds in yogurt are produced from lipolysis of milkfat and microbiological transformations of lactose and citrate. More than 100 volatiles, including carbonyl compounds, alcohols, acids, esters, hydrocarbons, aromatic compounds, sulfur-containing compounds, and heterocyclic compounds, are found in yogurt at low to trace concentrations. Besides lactic acid, acetaldehyde, diacetyl, acetoin, acetone, and 2-butanone contribute most to the typical aroma and flavor of yogurt. Extended storage of yogurt causes off-flavor development, which is mainly attributed to the production of undesired aldehydes and fatty acids during lipid oxidation. Further work on studying the volatile flavor compounds-matrix interactions, flavor release mechanisms, and the synergistic effect of flavor compounds, and on correlating the sensory properties of yogurt with the compositions of volatile flavor compounds are needed to fully elucidate yogurt aroma and flavor.     

SPME-GC-MS结合ROAV分析单菌及复配发酵牛乳中关键性风味物质

采用固相微萃取-气相色谱-质谱联用技术检测分析Streptococcus thermophilus与Lactobacillus delbrueckii subsp.bulgaricus单菌及复配发酵牛乳中的挥发性风味物质,结合相对气味活度值(relative odor activity value,ROAV)探讨发酵牛乳中关键性风味物质。结果表明:本实验共鉴定出100种挥发性风味物质,包括酸类、酮类、醛类、醇类、酯类、烷烃类和芳香族类化合物等。主成分分析表明,表征S.thermophilus单菌发酵乳的关键性风味物质是双乙酰、正壬醛和甲苯;表征L.bulgaricus单菌发酵乳的关键性风味物质是正庚醛、丁酸-2-甲基丙酯和1-庚醇;表征S.thermophilus与L.bulgaricus复配发酵乳的关键性风味物质是乙醛、3-甲基正丁醛、乙偶姻、2-壬酮、2-庚酮、醋酸乙烯酯、碳酸庚基苯基酯、甲酸乙烯酯和2-壬醇。相较于单菌发酵,复配发酵的风味物质组成、各组分相对含量及关键性风味物质均发生改变。     

Characterization of volatile compounds in fermented milk using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry

Lactic acid bacteria (LAB) are industrially important bacteria that are widely used in the fermented food industry, especially in the manufacture of yogurt. Characteristic flavors are produced by LAB during fermentation and storage that affect the quality and acceptability of fermented milk products. In this study, the volatile compounds in milk fermented by Streptococcus thermophilus IMAU80842 alone, Lactobacillus delbrueckii ssp. bulgaricus IMAU20401 alone, or both species together were identified using solid-phase microextraction methods coupled with gas chromatography-mass spectrometry. A total of 53, 43, and 32 volatile compounds were identified in milk fermented by S. thermophilus alone, L. delbrueckii ssp. bulgaricus alone, or both species together, respectively. The presence of some important flavor compounds was confirmed: acetic acid, acetaldehyde, acetoin, 2,3-butanedione, ethanol, and 1-heptanol. Our results demonstrate that the composition of the volatile compounds in fermented milk depends on the species of LAB used and whether they are used alone or in combination. This is important for the selection of appropriate starter cultures for the production of different types of fermented milk product with particular flavors.     

发酵乳风味及其分析技术研究进展

发酵乳制品具有悠久的历史, 富含蛋白质、脂肪、糖类和矿物质等营养物质, 具有调节机体功能、预防和治疗疾病、延长寿命等保健功能功效, 此外因其具有独特的发酵风味及组织状态, 深受消费者欢迎。发酵乳风味是评估其品质的主要指标之一, 乳酸菌在发酵过程中可以生成多种风味化合物, 其中主要包括酸类、酮类、酯类、醛类, 这些化合物的种类和含量均会影响发酵乳的品质?本文主要概述了发酵乳中挥发性风味物质的组成及形成机理, 同时对发酵乳关键风味化合物的分析技术进行了总结, 为发酵乳风味及其品质提高、新产品开发及关键技术工艺升级提供理论基础与依据。     

Probiotic fermented beverages based on acid whey

Acid whey is a byproduct of cheesemaking that is difficult to use because of its low pH and less-favorable processing properties compared with rennet whey. The aim of this study was to evaluate the qualities of fermented beverages made using acid whey. In manufacturing the beverages, we used probiotic cultures Lactobacillus acidophilus LA-5 or Bifidobacterium animalis ssp. lactis BB-12. The production process included combining pasteurized acid whey with UHT milk, unsweetened condensed milk, or skim milk powder. We introduced milk to enrich casein content and obtain a product with characteristics similar to that of fermented milk drinks. The products were stored under refrigerated conditions (5 ± 1°C) for 21 d. During storage, we assessed the beverages' physicochemical properties and organoleptic characteristics. The properties of the beverages depended on their composition, microbial culture, and storage time. Beverages containing L. acidophilus had higher acidity, which increased during storage; the acidity of samples containing B. animalis was more stable. Beverages made with skim milk powder (La1 and Bb1) had higher acetaldehyde content, but this parameter decreased in all samples during storage. The hardness of the samples did not change during storage and was highest in beverage La3, made from whey, condensed milk, and L. acidophilus. Beverage La2, made from whey, milk, condensed milk, and L. acidophilus, had the best sensory properties. The whey beverages we developed provided a good medium for the probiotic bacteria; bacteria count throughout the storage period exceeded 8 log cfu/mL, distinctly higher than the minimum therapeutic dose.