Characterization of Headspace Volatile Flavor Compounds Formed During Kefir Production: APplication of Solid Phase MicroextractioN

Headspace volatile flavor compounds of kefir were monitored using headspace solid phase microextraction (HS-SPME) method during fermentation of milk with kefir starter culture. Among all flavor compounds, forty volatile compounds were initially detected using gas chromatography coupled to time-of-flight mass spectrometer (GC-TOFMS). Consequently, eight volatile flavor compounds, namely ethanol, ethyl acetate, ethyl butyrate, 2-butanone, acetone, 3-hydroxy-2-butanone (acetoin), 2,3-butanedione (diacetyl) and acetaldehyde were considered as the representative of the alcohol, ketone, ester and aldehyde compounds in kefir. Moreover, in term of quantitative analysis, more than 97% of total flavor compounds composed of the proposed volatile flavor compounds. The results indicated that the concentration of 2-butanone released into headspace of kefir was found to be stable during fermentation. The release content of other volatile flavor compounds increased throughout the fermentation process. However, the headspace concentration of acetoin significantly (P < 0.05) decreased between pH 5.2 and 4.6.     

THE INFLUENCE OF 2-ACETOHYDROXY ACIDS ON THE DETERMINATION OF VICINAL DIKETONES IN BEER AND DURING FERMENTATION

During sampling and determination of diacetyl, 2-acetohydroxy acids are easily converted to vicinal diketones. A simple procedure for gas chromatographic determination of diacetyl, 2-acetolactate, acetoin and the homologous compounds is given. By careful sampling, less than 0·01 ppm of diacetyl was detected during the main fermentation in one brewery, whereas another strain of brewer's yeast yielded a maximum of 1·7 ppm of diacetyl. When samples of fermenting liquids are exposed to air at 60°C, complete conversion of 2-acetohydroxy acids takes place in less than one hour. The possibility that part of 2-acetolactate may be converted to acetoin, however, cannot be excluded. In finished beer 2-acetolactate levels of 0·2–0·5 ppm were observed. During the main fermentation the values ranged from 0·5–2·5 ppm.     

两种酸奶后酸化期风味物质的比较研究

比较了传统双菌发酵酸奶和高产γ-氨基丁酸的植物乳杆菌（Lactobacillus plantarum）与双菌复配发酵酸奶在后酸化过程中pH、双乙酰、乙醛及4种有机酸（柠檬酸、丙酮酸、乳酸、甲酸）的变化情况。试验结果显示,以上两种酸奶在后酸化期双乙酰和乙醛含量基本保持一致而有机酸含量差异较大。三菌复配型酸奶中甲酸含量较高而柠檬酸含量较低,其它两种有机酸在两种酸奶中的含量比较接近。上述结果表明,以德式乳杆菌保加利亚亚种（Lb.delbrueckiisubsp.bulgaricus）和嗜热链球菌（Streptococcus thermophilus）复配一定比例高产γ-氨基丁酸的植物乳杆菌为发酵剂制作的酸奶保持了酸奶原有的醇香,还形成了一定独特的风味特点。     

Simultaneous Determination of Sugars and Organic Acids in Cheddar Cheese by High-Performance Liquid Chromatography

In a simple, rapid isocratic HPLC method sugars and organic acids were separated on an Aminex HPX-87 column in the H⁺ form and detected using ultraviolet and refractive index detectors in series. Sugars (lactose, glucose and galactose) and acids (orotic, citric, pyruvic, lactic, uric, formic, acetic, propionic, butyric and hippuric) were identified by retention times. This method affords a simple technique for monitoring starter culture activity and following quality changes during cheese maturation.     

具有良好风味嗜热链球菌的筛选及其产香特性分析

嗜热链球菌作为酸奶发酵剂的常用菌株之一,在牛乳发酵和贮藏过程中可以赋予产品优良的质地、丰富的营养价值和独特的风味。本实验在前期研究的基础上,以商业发酵剂为对照组,以分离自不同地区传统发酵乳中的具有良好发酵特性的7株嗜热链球菌为实验菌株,采用固相微萃取结合气相色谱-质谱联用技术对发酵终点,即pH4.5时发酵乳中的挥发性风味物质进行检测分析。结果表明,在所有实验菌株中,G80-2发酵乳中的挥发性风味化合物的组成及含量最接近对照组。在此基础上,对G80-2菌株在牛乳发酵和贮藏过程中所产生的挥发性风味物质进行动态分析,发现该菌株在发酵和贮藏期间检测出酸类（4种）、醇类（10种）、酮类（12种）、醛类（6种）、酯类（1种）等多种化合物,且一些主要特征风味物质如乙酸、乙醛、双乙酰、乙偶姻、2-庚酮、1-庚醇等相对含量较高,说明该菌株可作为发酵剂应用于乳制品生产中。     

Microbiological and fermentation characteristics of togwa,a Tanzanian fermented food

Selected microbiological and metabolic characteristics of sorghum, maize, millet and maize-sorghum togwa were investigated during natural fermentation for 24 h. The process was predominated by lactic acid bacteria (LAB) and yeasts. The mesophiles, lactic acid bacteria, and yeasts increased and the Enterobacteriaceae decreased to undetectable levels within 24 h. The isolated microorganisms were tentatively identified as Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus fermentum, Lactobacillus cellobiosus, Pediococcus pentosaceus, Weissella confusa, Issatchenkia orientalis, Saccharomyces cerevisiae, Candida pelliculosa and Candida tropicalis. The pH decreased from 5.24-5.52 to 3.10-3.34. Maltose increased initially and then decreased, fructose decreased and glucose levels increased during the first 12 h of fermentation. The organic acids detected during fermentation included DL-lactic, succinic, formic, pyruvic, citric, pyroglutamic and uric acid. Lactate was the predominant acid and increased significantly with time. The volatile organic compounds (VOC) detected included acetaldehyde, 2-methyl-propanal, 2-methyl-butanal, 3-methyl-butanal, ethanol, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, diacetyl and acetoin. Ethanol was the predominant VOC and it increased significantly with time.     

High Performance Liquid Chromatographic Determination of Organic Acids in Dairy Products

A simple isocratic HPLC technique was developed for the quantitative analysis of organic acids in dairy products. An Ammex HPX- 87 column at 65°C, 0.0090N H₂SO₄ mobile phase and UV detection at 220 and 275 nm were utilized. Orotic, citric, pyruvic, lactic, uric, formic, acetic, propionic, butyric, and hippuric acids were quantitated for whole milk, skim powder, cultured buttermilk, sour cream, cottage cheese, yogurt, sharp Cheddar cheese, and blue cheese. Over 90% recoveries of acids added to whole milk were observed for ail acids except butyrid; the average recovery for butyric was 86%.     

Volatile flavour compounds of yoghurt

The volatile flavour compounds of three samples of Egyptian yoghurt were analysed over a two-week period at 8°C using a simple headspace gas chromatography technique in order to study the changes and relate them to flavour acceptability.
Volatile compounds present were acetaldehyde, diacetyl, acetoin, acetone, butanone, and acetic acid. Acetone and butanone disappeared within the first week of storage, whereas acetaldehyde, diacetyl and acetoin declined steadily but were still present after two weeks. Acetic acid increased to about twice its original level after 10d of storage.
The decreases in acetaldehyde, diacetyl, acetoin, and the increase of acetic acid were closely related to the rapid decrease in product acceptability after 8–10d storage.     

发酵乳风味物质乙醛、双乙酰的合成途径及其调控机制

乳酸菌在发酵过程中能够产生醇、醛、酮等多种风味物质,这些风味物质和乳酸菌在糖酵解过程中产生的乳酸、生醋酸、丙酸等有机酸相互作用,形成具有独特风味的发酵乳。在酸奶的风味物质中,乙醛、双乙酰是其主要成分。目前,关于酸奶风味物质研究的焦点主要集中在通过改变乙醛、双乙酰的代谢途径提高乙醛、双乙酰的产量上。这篇综述主要描述了乳酸菌在酸奶发酵过程中生产的风味物质、以及乙醛、双乙酰的合成途径和国内外学者在乙醛、双乙酰产量调控方面取得的研究成果。     

Diacetyl levels and volatile profiles of commercial starter distillates and selected dairy foods

Starter distillates (SDL) are used as ingredients in the formulation of many food products such as cottage cheese, margarine, vegetable oil spreads, processed cheese, and sour cream to increase the levels of naturally occurring buttery aroma associated with fermentation. This buttery aroma results, in part, from the presence of the vicinal dicarbonyl, diacetyl, which imparts a high level of buttery flavor notes and is a key component of SDL. Diacetyl (2,3-butanedione) is a volatile product of citrate metabolism produced by certain bacteria, including Lactococcus lactis ssp. diacetylactis and Leuconostoc citrovorum. In the United States, SDL are regarded as generally recognized as safe ingredients, whereby usage in food products is limited by good manufacturing practices. Recently, diacetyl has been implicated as a causative agent in certain lung ailments in plant workers; however, little is published about the volatile composition of SDL and the levels of diacetyl or other flavoring components in finished dairy products. The objective of this work was to characterize the volatile compounds of commercial SDL and to quantitate levels of diacetyl and other Flavor and Extract Manufacturers Association-designated high-priority flavoring components found in 18 SDL samples and 24 selected dairy products. Headspace volatiles were assessed using a solid-phase microextraction and analyzed by gas chromatography-mass spectrometry. In addition to diacetyl (ranging from 1.2 to 22,000 μg/g), 40 compounds including 8 organic acids, 4 alcohols, 3 aldehydes, 7 esters, 3 furans, 10 ketones, 2 lactones, 2 sulfur-containing compounds, and 1 terpene were detected in the SDL. A total of 22 food samples were found to contain diacetyl ranging from 4.5 to 2,700 μg/100g. Other volatile compounds, including acetaldehyde, acetic acid, acetoin, benzaldehyde, butyric acid, formic acid, furfural, 2,3-heptanedione, 2,3-pentanedione, and propanoic acid, were also identified and quantified in SDL or food samples, or both. The results obtained in this work summarize the volatile composition of commercial SDL and the approximate levels of diacetyl and other Flavor and Extract Manufacturers Association-designated high-priority flavoring components found in SDL and selected dairy foods.     

Determination of several flavours in beer with headspace sampling–gas chromatography

In order to determine flavours (diacetyl, pentanedione, acetoin and acetaldehyde) in beer, gas chromatography coupled with headspace sampling technique was developed in our laboratory. The calibrations were accomplished by six level standard addition methods, and relative coefficients were ?0.999. Repeatability experiments were accomplished by detecting 20 successive same standard samples, which showed that detection precision was <2% relative standard deviation (RSD) for three flavours (diacetyl, pentanedione and acetaldehyde), and detection precision of acetoin was <2.5% (RSD). Four flavours gradually increased and then decreased during the process of fermentation, and they leveled off when the fermentation was over. The ratio (diacetyl/pentanedione) reflected the degree of contamination: normal beer had a ratio of approximate 1; the ratio (diacetyl/pentanedione) was higher than 1, displaying that beer was contaminated by microbes. Our data suggested that gas chromatography coupled with headspace sampling technique could be used successfully to determine flavours (diacetyl, pentanedione, acetoin and acetaldehyde) in beer.     

Effects of frozen and refrigerated storage on organic acid profiles of goat milk plain soft and Monterey Jack cheeses

The effects of 6 mo of freezing and refrigeration on organic acid profiles of 2 types of goat milk cheese [plain soft (PS) and Monterey Jack (MJ)] were studied in comparison with those of a nonfrozen control (NFC). Three lots of commercial PS cheeses were purchased, and 3 lots of MJ cheeses were manufactured at the University dairy plant. Each lot of the 2 types of cheeses was subdivided into 4 equal portions, and one subsample of each cheese was immediately stored at 4°C as the NFC for 0, 14, and 28 d. The other 3 were immediately frozen (−20°C) for 0, 3, and 6 mo (0MF, 3MF, and 6MF) and subsequently thawed the next day at 4°C. The samples were then stored at 4°C for 0, 14, and 28 d. Organic acids were quantified using an HPLC. The PS had no pyruvic acid, and MJ contained no isotartaric acid; however, several unknown large peaks appeared between propionic and butyric acids. Differences in organic acid contents between PS and MJ cheeses were significant for all acids except citric and lactic acid. Lot effect was significant for most of the known acids, indicating that variations existed in milk composition and manufacturing parameters. Effects of storage treatments (NFC, 0MF, 3MF, and 6MF) were significant for most organic acids, except for orotic and a few unidentified acids. Aging at 4°C for 4 wk had little influence on all organic acids, except butyric acid. Concentrations of butyric, lactic, propionic, tartaric, and uric acids were significantly elevated as the frozen storage period advanced. At the initial stage, there were no differences in pH and acid degree values between NFC and frozen-stored groups of both cheeses. However, acid degree values gradually increased as the refrigerated storage extended up to 4 wk, indicating that lipolysis increased as the refrigeration storage at 4°C advanced. Although levels of several organic acids were changed in the goat cheeses, the prolonged frozen storage, up to 6 mo, was apparently feasible for extending storage.     

CHANGES IN THE PROFILE OF ORGANIC ACIDS IN PLAIN SET AND STIRRED YOGURTS DURING MANUFACTURE AND REFRIGERATED STORAGE1

The organic acids profile in fermented dairy foods is an indicator of the metabolic activity of added bacterial cultures. These acids act as natural preservatives and contribute to the characteristic sensory properties. This study was done to determine the effect of encapsulation on the metabolic activity of probiotic supplements, i.e. bifidobacteria, in plain yogurts. Metabolically active bifidobacteria may increase the concentration of certain organic acids, mainly acetic and propionic acids, which might lead to decreased acceptability of the product. Changes in the profiles of organic acids were determined in plain set and stirred‐type yogurts containing the starter culture and microencapsulated and nonencapsulated probiotic strains of either Bifidobacterium longumB6 or B. longum ATCC15708. Ion‐exchange high‐performance liquid chromatography was used for the separation and quantification of the organic acids. Concentrations of acetic and lactic acid increased during storage, while those of citric and uric acid remained stable. No particular pattern was observed for propionic or butyric acid, while pyruvic acid initially decreased and then increased during storage.     

藏灵菇发酵奶发酵特性的研究

本文研究了藏灵菇发酵奶发酵过程中酸度、有机酸、挥发性风味物质及微生物数量的变化,在发酵0、4、8、12、16、20h时采样,用高效液相色谱(HPLC)跟踪发酵过程中乳酸、乙酸和柠檬酸的变化;用气相色谱(GC)跟踪了发酵过程中乙醇、丁二酮、乙醛和丙酮的变化。实验结果表明,乳酸和乙酸在发酵过程中都呈上升趋势,发酵4h时,乳酸的生成量约是乙酸的20倍,发酵结束乳酸的量为8.56g/L(TK1),7.73g/L(TK2);乙酸的量为0.71g/L(TK1),0.81g/L(TK2),乳酸的量约是乙酸的9～12倍。柠檬酸从发酵开始就呈下降趋势。挥发性风味物质中乙醇的生成量最大,发酵结束分别达到3134mg/L(TK1),4994mg/L(TK2);丁二酮和乙醛的量较普通酸奶高,分别达到62.4mg/L(TK1)和37.8mg/L(TK1)。本研究也对发酵过程中微生物进行了分析,发酵结束,发酵奶中乳杆菌达到2.3×109CFU/ml(TK1),1.3×108CFU/ml(TK2);乳球菌达到1.3×109CFU/ml(TK1),1.1×108CFU/ml(TK2);酵母菌1.6×106CFU/ml(TK1),6.6×106CFU/ml(TK2)。TK1和TK2之间存在较大差异,TK1的风味好于TK2。     

Changes in organic acids during ripening of cheeses made from raw, pasteurized or high-pressure-treated goats’ milk

Organic acids of cheeses made from raw (RA), pasteurized (PA; 72°C, 15 s) or pressure-treated (PR; 500 MPa, 15 min, 20°C) goats’ milk were qualitatively and quantitatively assessed during ripening. Nine organic acids (citric, pyruvic, malic, lactic, formic, acetic, uric, propionic and butyric) were analysed in each sample by HPLC.Milk treatment did not affect the total organic acids content of 1-day-old cheeses, which increased steadily from day 1 to day 60. At the end of ripening, RA and PR milk cheeses both exhibited higher concentration of organic acids than in those made from PA milk.Lactic acid was found in higher concentration in PR milk cheese from 30 days of ripening. The RA milk cheese, that showed the highest nonstarter lactic acid bacteria counts, were characterized by an elevated amount of propionic and acetic acids. These cheeses also were negatively correlated with both pyruvic and citric acid contents. The PA milk cheese showed a high level of malic acid, and was clearly differentiate from RA and PR milk cheeses by its low level of butyric acid.     

Evaluation of volatile compounds and free fatty acids in set types yogurts made of ewes’, goats’ milk and their mixture using two different commercial starter cultures during refrigerated storage

Six different types of yogurt were manufactured from Damascus goat milk, Awassi ewe milk and a mixture of equal portions of the 2 species of milk using 2 types of commercial yogurt cultures (CH-1 and YF-3331). Yogurts were chemically analysed at 1, 7, 14 and 21 days of storage. Results showed that cultures significantly affected acetaldehyde (P < 0.05), acetone (P < 0.05) and diacetyl (P < 0.001) contents. Type of milk significantly influenced acetaldehyde (P < 0.05), diacetyl (P < 0.001), acetoin (P < 0.001) and ethanol (P < 0.05) levels. Significant variations occurred in acetaldehyde (P < 0.001) and acetoin (P < 0.05) contents during the storage. Short-chain free fatty acids were the highest in ewes’ milk yogurt made with culture YF-3331, and increased during storage, while the levels of medium-chain free fatty acids, except for decanoic acid, were unchanged and the amount of long-chain free fatty acids decreased during storage. Cultures used and types of milk had no effect on long-chain free fatty acids in yogurts.     

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.     

Citrate utilization in milk by Leuconostoc cremoris and Streptococcus diacetilactis.

Citrate utilization and diacetyl, acetoin and acetaldehyde production by 2 strains each of Leuconostoc cremoris and Streptococcus diacetilactis in milk were studied. With the leuconostoc bacteria no growth and little citrate utilization occurred unless a stimulant (yeast extract) was present, when complete utilization of citrate without concomitant production of diacetyl or acetoin was obtained. The additon of Mn2+ stimulated growth resulted in diacetyl and acetoin production. Destruction of diacetyl and acetoin occurred when the citric acid level fell to c.1000 and 600 mug/g in the case of Leuc. cremoris FR8-1 and CAF1, respectively. Only strain FR8-1 produced acetaldehyde. In contrast, Str. diacetilactis produced diacetyl, acetoin and acetaldehyde concomitant with citrate utilization.     

雪里蕻腌菜风味物质的研究

利用GC-MS技术对雪里蕻腌菜卤汁乙醚萃取液中组分进行了鉴定分析,结果鉴定出有机酸组分8个,醇类组分4个,腈类组分2个,酯类组分1个,芳烃类组分2个,并对卤汁中的醇类组分乙醇、丙醇、丁醇及2,3-丁二醇进行了气相色谱检测,卤汁中的乙醛、乙偶姻、双乙酰经2,4*二硝基苯肼衍生化后进行了高效液相色谱分析,还对其中重要风味组分的乳酸发酵机理及其对雪里蕻腌菜风味的影响进行了分析讨论。     

发酵条件对酸樱桃发酵汁挥发性风味物质的影响探究

目的 考察发酵条件对酸樱桃发酵汁中风味物质的影响。方法 利用3株乳酸菌发酵酸樱桃汁, 以pH为指标明确最佳发酵菌种和稀释比例。随后利用气相色谱-离子迁移谱法(gas chromatography-ion mobility spectrometry, GC-IMS)探究糖添加量、pH、氮源、稀释比例对发酵挥发性风味物质的影响。结果 最佳发酵菌为鼠李糖乳杆菌(Lactobacillus GG, LGG), 最佳稀释比例为1:3, 氮源对挥发性风味物质几乎没有影响; 而不同添加量的碳源(葡萄糖)导致发酵液乙偶姻、双乙酰等含量差异; 随着pH增加, 乙偶姻、双乙酰、2-戊酮等含量显著升高; 稀释比例通过影响发酵原汁的浓度导致发酵后风味物质乙偶姻含量显著增加、2-戊酮含量显著降低。进一步差异比对3株乳酸菌酸樱桃发酵汁的风味, 发现LGG代谢脂肪酸可产生较多的2-庚酮及2-戊酮。结论 菌种、糖添加量、稀释比例、起始pH均可导致发酵液中乙偶姻、2-庚酮及2-戊酮等挥发性风味物质产生显著影响, 本研究为酸樱桃的加工利用提供理论指导。