Histamine formation by histamine-forming bacteria in douchi,a Chinese traditional fermented soybean product

Seven soybean and 19 black bean douchi products sold in the supermarkets in southern Taiwan were purchased and tested to determine the occurrence of histamine and histamine-forming bacteria. The levels of pH, salt content, water content, yeast and mold, and aerobic plate count (APC) in all samples ranged from 4.7 to 5.9, 4.4% to 14.0%, 6.8% to 51.6%, 3.0 to 5.1 log CFU/g, and 5.2 to 9.2 log CFU/g, respectively. None of these samples contained total coliform and Escherichia coli. Although black bean douchi products had an average histamine content of 29.0 mg/100 g, 18 of them had histamine contents greater than 5 mg/100 g, the allowable level set by the US Food and Drug Administration (FDA) for scombroid fish and/or products. In contrast, only four soybean douchi products had histamine levels greater than 5 mg/100 g. Among the black bean samples, four contained histamine at 56.3, 62.1, 80.2 and 80.8 mg/100 g, that are above the 50 mg/100 g hazard action level. Eight histamine-forming bacterial strains, capable of producing 11.7–601 ppm of histamine in trypticase soy broth (TSB) supplemented with 1% l-histidine (TSBH), were identified as Bacillus subtilis (four strains) Staphylococcus pasteuri (one strain) and Staphylocuccus capitis (three strains) by 16S rDNA sequencing with PCR amplification. S. capitis, which was previously reported to be halotolerant, was a potent histamine-former capable of producing more than 500 ppm of histamine in TSBH in the presence of 0.5–10% NaCl.     

Angiotensin I-converting enzyme inhibitory peptides in douchi,a Chinese traditional fermented soybean product

Douchi, a soybean product originating in China, produces angiotensin I-converting enzyme (ACE) inhibitors with the potential to lower blood pressure. The ACE inhibitory activities of douchi qu pure-cultured by Aspergillus Egyptiacus for 48 h, and 72 h were compared with douchi secondary-fermented for 15 d. The results showed that ACE inhibitory activities were improved following the fermentation. ACE inhibitory activities of 48 h-primary-fermented douchi qu did not change dramatically after preincubation with ACE, but increased greatly after preincubation with gastrointestinal proteases. The results suggest they were pro-drug-type or a mixture of pro-drug-type and inhibitor-type inhibitors. The ACE inhibitors in 48 h-fermented douchi qu were fractionated into four major peaks by gel filtration chromatography on Sephadex G-25. Peak 2, which had the highest activity, had only one peptide, composed of phenylalanine, isoleucine and glycine with a ratio of 1:2:5.     

Isolation and characterisation of a novel angiotensin I‐converting enzyme‐inhibitory peptide derived from douchi,a traditional Chinese fermented soybean food

BACKGROUND: Douchi, a traditional fermented soybean food, has recently attracted a great deal of attention owing to its superior physiological activity. In the present study the angiotensin I‐converting enzyme (ACE)‐inhibitory activity of typical douchi procured from various regions of China was analysed. An ACE‐inhibitory peptide derived from the most potent douchi was also isolated and characterised. The pattern of ACE inhibition and resistance to hydrolysis by gastrointestinal proteases of this peptide are described. RESULTS: ACE‐inhibitory activities were detected in all douchi samples, with IC₅₀ values ranging from 0.204 to 2.011 mg mL^?1. Among the douchi samples, a Mucor‐type douchi exhibited the most potent ACE‐inhibitory activity (IC₅₀ = 0.204 mg mL^?1). A novel ACE‐inhibitory peptide was then isolated from this Mucor‐type douchi using ultrafiltration followed by Sephadex G‐25 column chromatography and reverse phase high‐performance liquid chromatography. The amino acid sequence of the purified peptide was identified by Edman degradation as His‐Leu‐Pro (IC₅₀ = 2.37 µmol L^?1). The peptide is a competitive inhibitor and maintained its inhibitory activity even after incubation with some gastrointestinal proteases. CONCLUSION: The present study shows that peptides derived from soybean fermentation during douchi processing could be the main contributor to the ACE‐inhibitory activity observed. Copyright © 2009 Society of Chemical Industry     

Volatile compounds characterising Tunisian virgin olive oils: the influence of cultivar

In this work, the volatile profiles of four Tunisian virgin olive oils were established by headspace‐solid phase microextraction and gas chromatography, using flame ionisation and mass spectrometer detectors. More than eighty compounds were isolated and characterised, representing 97–99% of the total GC area. The main volatile compounds present in the oil samples were determined quantitatively, the major constituents were often aldehydes, particularly (E)‐2‐hexenal (48–90%). Significant differences in the content of volatile constituents were observed. These quantitative differences were used to distinguish between virgin olive oils from different varieties.     

Volatile components of the enzyme-ripened sufu, a Chinese traditional fermented product of soy bean

In the present study, sufu, a soft cheese-like oriental fermented food, was prepared by ripening the salted-tofu cubes in Aspergillus oryzae-fermented soybean-rice koji at 37°C for 16 days (16-day sufu). Sufu was further held at room temperature for another 30 days (46-day sufu). The volatile components of the non-fermented tofu cubes and the sufu products were identified and quantified by GC and GC-MS. A total of 70 volatile compounds including 20 aldehydes, 18 alcohols, 16 esters, 5 ketones, 5 acids and 6 other compounds were identified. Sufu products contained more volatile compounds than non-fermented tofu cubes qualitatively and quantitatively. After 16-days of ripening, fatty acid, aldehyde and ester were noted to be the dominant volatile fractions. In contrast, the 46-day sufu contained ester, and alcohol as the major volatile fractions. They comprise approximately 63.9% of the total volatile components.     

Effect of fermented rice culture on the microbiological,biochemical and sensory characteristics of low‐salt douchi,a traditional Chinese fermented soybean condiment

A low‐salt (4% NaCl, w/w) douchi supplemented with different concentration of fermented rice culture (FRC) of 0%, 5%, 10%, 15% and 20% (v/w) was produced, and the effect of FRC on microbiology, biochemistry and sensory quantity was investigated. Results indicated that FRC had a significant effect on low‐salt douchi. Compared with the control (added 0% FRC), in the other treatments, the counts of yeast and lactic acid bacteria increased by about 50%, whereas the moulds decreased by about 30%; the pH reduction was close to 4.7; total acid increased to about 1.300 g per 100 g; reducing sugar maintained a decrease to 21.07 mg per g; amino nitrogen increased to about 0.900 g per 100 g, but not above the control (1.013 g per 100 g). In general terms, total free amino acid contents and three organic acids were higher in low‐salt products than four commercial products. Sensory evaluation of products supplemented with 10% and 15% FRC revealed better overall acceptability compared with other treatments and a commercial product.     

Changes in volatile flavour compounds in field pea cultivars as affected by storage conditions

The impact of storage conditions on volatile flavour profile of field pea cultivars was evaluated. Seeds were kept at 4 °C, room temperature (approximately 22 °C) and 37 °C for 12 months. Headspace solid‐phase microextraction gas chromatography–mass spectrometry was used for extraction and identification of volatile flavour compounds (VFCs). Significant (P < 0.01) differences in the concentration of VFCs were observed during storage. All cultivars kept at 22 and 37 °C had higher mean value of aldehydes compared to those kept at 4 °C. Alcohols, hydrocarbons, ketones, terpenes and esters were higher in samples stored at 4 °C compared to those at higher temperatures. 1‐Hexanol, hexanal, styrene, 2‐butanone, dimethyl sulphide, 3‐carene, ethyl acetate and 2, 3‐diethyl‐5‐methyl pyrazine were the most abundant compounds found in peas. Results from this study could be useful in identifying improved conditions of storage to enhance flavour properties of peas.     

基于不同发酵方法制作的曲拉中挥发性风味物质分析

采用顶空-固相微萃取（HS-SPME）方法对传统自然发酵的曲拉与商品发酵剂制作的曲拉样品进行预处理,利用气相色谱-质谱法（GC-MS）检测样品中的挥发性风味成分。经NIST14质谱数据库检索结合文献对照,从传统自然发酵的曲拉中检出29种挥发性风味成分,商品发酵剂制作的曲拉中检出53种,这些挥发性成分中主要包括酸类、醇类、酮类、醛类、酯类、烃类、芳香类。不同发酵方法制作的曲拉中风味物质的种类及含量都存在一定的差异性,这也是使用相同原料不同发酵方法制作的曲拉产生特别的风味的主要原因。      

不同蔬菜原料发酵泡菜挥发性成分解析

为探明不同原料发酵泡菜特征性挥发性成分及其差异,采用顶空固相微萃取-气相色谱质谱联用法(HS-SPME-GC-MS)对6 种典型泡菜(泡生姜、泡长萝卜、泡圆萝卜、泡青芥菜、泡榨菜、泡蒜)中的挥发性成分进行检测。主成分(PCA)分析表明,泡菜的主要挥发性成分根据发酵程度不同可以分为两大类。一类为充分发酵的蔬菜,挥发性成分主要来自于发酵和原料本身,如萝卜、青芥菜、榨菜发酵后产生的乙醇、乙酸和酯类等,也包括原料自身的硫醚类、异硫氰酸烯丙酯等。另一类为不充分发酵的蔬菜,挥发性成分主要来自本身,如泡蒜主要挥发性成分为二烯丙基二硫醚等。发酵型蔬菜中榨菜、青芥菜和萝卜发酵后风味差异较大,即便是同类型不同品种如长萝卜和圆萝卜,发酵后挥发性成分也有较大差异。乙醇、苯乙醇、2-乙基己醇、乙酸、丁酸、乙酸乙酯可以用于区分发酵型和不发酵型泡菜,其中苯乙醇和2-乙基己醇可以用来区分泡长萝卜和泡圆萝卜。     

固相微萃取条件优化及发酵菜粕风味物质分析

研究固相微萃取与气质联用研究发酵菜粕的风味物质及比较不同品种菜粕发酵前后风味物质的变化,以峰面积及峰数为指标,通过萃取头、萃取温度、吸附时间和样品量的优化,确定了固相微萃取和气质联用研究发酵菜粕风味固相微萃取的条件,并比较了3种代表性菜粕发酵前后风味物质及硫苷组分和含量的变化。结果表明,固相微萃取最佳条件是:50/30μm DVB/CAR/PDMS(DCP)萃取头,温度70℃,吸附30 min,样品量2 g,其中萃取头和吸附温度显著影响萃取效率。发酵前后菜粕的挥发性组分及含量变化都较大,发酵后风味物质的总量都显著增加,硫苷的含量明显降低,表明发酵不但显著增加了风味物质的含量改变了风味,而显著降低了抗营养物质硫苷的含量。     

Optimization of solid‐phase microextraction methods for GC‐MS determination of terpenes in wine

Solid‐phase microextraction using a 100 µm poly(dimethylsiloxane) fiber, followed by gas chromatography–mass spectrometry determination, has been optimized for the analysis of some terpenes in wine samples. The best results were obtained by direct immersion of the fiber using a sampling period of 15 min with constant magnetic stirring (1100 rpm) and an extraction temperature of 20 °C. The sample volume was 7 ml with 25% NaCl, in a 15 ml capped vial. Desorption was performed directly in the gas chromatograph injector port over 5 min at 250 °C using the splitless mode. The method is sensitive, with detection limits between 11 and 25 µg l^?1, precise, with variation coefficients in the range 1.28–3.71%, and linear over more than one order of magnitude. The related conditions were used for wine sample analyses with recoveries between 71.8 and 90.9%. Solid‐phase microextraction remains an attractive alternative technique due to its rapidity and because it is a solvent‐free extraction. Copyright © 2005 Society of Chemical Industry     

SDE和SPME法对鲊海椒发酵中香气组成的比较分析

为确定评价鲊海椒挥发性香气组成的最优方法,以粳米粉为淀粉原料制备鲊海椒,采用固相微萃取（solidphase microextraction,SPME）法和同时蒸馏萃取（simultaneous distillation extraction,SDE）法并结合感官评价比较研究其在发酵过程中风味物质的变化。结果发现,SPME法萃取发酵0、45 d和90 d鲊海椒挥发性成分种类分别为76、131 种和122 种,SDE法挥发性成分分别为23、42 种和82 种;随着发酵时间的延长,SDE法挥发性香气成分增加,而SPME法在发酵45 d时挥发性香气物质种类达峰值并保持稳定。酯类是鲊海椒发酵过程中主要的挥发性成分,采用SPME法和SDE法萃取发酵45～90 d鲊海椒,分别检出挥发性酯类42～50 种（相对含量46.7%～55.3%）和15～27 种（相对含量60.28%～67.28%）;SPME法检出较多醇类、醛类,发酵45～90 d鲊海椒醇类14～21 种（相对含量15.7%～22.6%）和醛类17 种（相对含量5.8%～12.3%）。SPME法可萃取得到更多低沸点、小分子化合物;而SDE法得到更多高沸点化合物;感官评定显示发酵45 d后的鲊海椒色泽鲜艳,具有特殊的酸味和醇香味。结果表明,采用SPME法结合SDE法评价鲊海椒挥发性香气组成更加全面客观,鲊海椒适宜发酵时间为45～90 d。     

Volatile compounds in light, medium, and dark black walnut and their influence on the sensory aromatic profile

Light, medium, and dark colored kernels from 3 different cultivars (Emma K, Kwik Krop, and Sparks 127) and one wild species of black walnut were studied for their aroma volatiles. Solid-phase microextraction-gas chromatography-mass spectrometry was used to determine the aromatic compounds present in the headspace of these samples. Partial least square regression was used to correlate the instrumental aromatic data with the sensory responses, obtained in a previous study, for the same samples. Thirty-four aromatic compounds were found in the black walnut cultivars, highlighting among them the presence of 14 esters. Although more than 50% of the total concentration of volatile compounds, esters were not important compounds in determining the differences in the sensory aromatic profiles of the 3 colors of the nuts. As a general trend, the concentration of total volatile compounds was always significantly higher in light black walnuts than in the medium colored samples; medium colored samples had higher volatile content than the dark black walnuts. The presence of hexanal was related to rancid and acrid aromas and was determined to differentiate the dark black walnuts from the medium and light colored samples. PRACTICAL APPLICATION: The data presented in this article will help understand the aromatic differences between light, medium, and dark colored kernels of domestic and wild black walnut. The aromatic profile of these nuts, not studied until this moment, can be used as a model to develop flavorings and new products by the food industries.     

Changes in volatile aroma compounds of pineapple (Ananas comosus) during freezing and thawing

Changes in the volatile aroma compounds of pineapple (Ananas comosus) during freezing and thawing were compared against fresh samples to determine the effect of freezing on pineapple flavour. An HS‐SPME–GC–MS analysis showed that the Smooth Cayenne pineapple variety had nineteen volatile compounds, in four classes of compounds including fourteen esters, two hydrocarbons, two sulphur‐containing compounds and one lactone. The main characteristic volatile compounds of the fresh pineapple were methyl hexanoate, ethyl hexanoate, ethyl 3‐methylthiopropanoate and 1‐(E,Z)‐3,5‐undecatriene. Freeze–thaw cycles were associated with the loss of some volatile aroma compounds, particularly the esters which were found to be the main characteristic of fresh pineapples. The freezing and thawing process was found to cause damage to the pineapple tissues due to ice recrystallisation and dehydration which lead to the reduction of volatile aroma compounds.     

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。     

Biodiversity and origin of the microbial populations isolated from Masske,a traditional Iranian dairy product made from fermented Ewe's milk

Masske is a traditional Iranian dairy product containing 50% butterfat made from fermented ewe's milk. Overall, 672 bacterial isolates were collected from ewe's milk, fermented milk (FM) and Masske samples that were produced in households located in southern regions of the Khorasan Province in Iran. To identify lactic acid bacteria in these samples, a total of 79 Gram‐positive and catalase‐negative isolates were analysed. The identification of isolates was achieved by phenotypic and sequential analysis of the 16S rRNA gene. Enterococcus faecium and Aerococcus viridans were the most frequently isolated species in the samples, but the most commonly present bacteria in Masske were Streptococcus thermophilus.     

1,3‐Dimethoxybenzene,a newly identified component of port wine

1,3‐Dimethoxybenzene was identified by GC–O, GC–MS and Kovats indices (polar Supelcowax, 1709; non‐polar Rtx‐5MS, 1158) as a new volatile component of port wine. Sensory evaluation described this compound as having a sweet medicinal odour with hazelnut, resinous and woody notes. Respective threshold limits in model wine and port wine were 21 and 47 µg l^?1. Quantitative analysis by GC–MS, using a selected characteristic ion (m/z 138), indicated that young port wines from the 1998 vintage contained up to 3 µg l^?1 whereas ports from the 1999 vintage contained up to 20 µg l^?1. © 2002 Society of Chemical Industry