Biogenic amines in low- and reduced-fat dry fermented sausages formulated with konjac gel

Biogenic amines in low- and reduced-fat dry fermented sausages made with konjac gel (KG) as pork backfat replacer were studied. An increase (P < 0.05) was observed in the microbial count during the fermentation process, reaching levels of over 8 Log cfu/g of total viable microorganisms and lactic acid bacteria. However, no significant differences were observed in the microbiota evolution as a function of the reformulation process (fat and konjac gel content). High levels of physiological amines (spermidine, spermine, and agmatine) were observed in the raw material. From day 2 of the fermentation process an increase (P < 0.05) was observed in tyramine and putrescine, which were the predominant amines at the end of the storage period. The increase in these amines was proportional to the presence of KG and fat reduction. This can also be seen for spermine, with agmatine showing the inverse. The biogenic amine levels in these products reformulated with KG are not considered to pose a health risk to consumers.     

Determination of biogenic amines in squid and white prawn by high-performance liquid chromatography with postcolumn derivatization

A simple method was developed for the determination of biogenic amines in aquatic food products using a reverse-phase high-performance liquid chromatography with postcolumn automatic o-phthalaldehyde derivatization and fluorescence detection. The linearity, repeatability, and recovery of the method for seven amines (tyramine, putrescine, cadaverine, histamine, agmatine, spermidine, and spermine) were evaluated. This optimized method was applied to detect biogenic amines in squid and white prawn during refrigerated storage. Sensory analysis, pH value, and total volatile base nitrogen value were combined to evaluate the freshness quality of these two raw materials. Agmatine and cadaverine in squid, cadaverine, and putrescine in white prawn were the most obviously changed amines during the storage at two different temperatures, and these biogenic amines could be the effective quality indicators for the freshness of the raw aquatic materials.     

Biogenic amine formation and nitrite reactions in meat batter as affected by high-pressure processing and chilled storage

Changes in biogenic amine formation and nitrite depletion in meat batters as affected by pressure-temperature combinations (300 MPa/30 min/7, 20, and 40 degrees C), cooking process (70 degrees C/30 min), and storage (54 days/2 degrees C) were studied. Changes in residual nitrite concentration in raw meat batters were conditioned by the temperature and not by the pressure applied. Cooking process decreased (P < 0.05) the residual nitrite concentration in all samples. High-pressure processing and cooking treatment increased (P < 0.05) the nitrate content. Whereas protein-bound nitrite concentration decreased with pressure processing, no effect was observed with the heating process of meat batters. High-pressure processing conditions had no effect on the rate of residual nitrite loss throughout the storage. The application of high pressure decreased (P < 0.05) the concentration of some biogenic amines (tyramine, agmatine, and spermine). Irrespective of the high processing conditions, generally, throughout storage biogenic amine levels did not change or increased, although quantitatively this effect was not very important.     

Biogenic amines in wines from three Spanish regions

One hundred and sixty-three wines from La Rioja, Utiel-Requena, and Tarragona were analyzed to determine if there were any differences in the concentrations of six biogenic amines that are found in these three regions. The influence of grape variety, type of vinification, wine pH, malolactic fermentation, and storage in bottle on biogenic amine concentrations was studied. Results show important differences in putrescine and histamine concentrations among regions, varieties of grape, and type of wine; differences were less appreciable for the remaining biogenic amines studied. Low pH prevented biogenic amine formation. Malolactic fermentation and short storage periods in bottle (3-6 months) showed increases in histamine concentration, whereas longer periods of storage led to a general decrease in histamine. Several strains of lactic acid bacteria were isolated in this work, and their ability to form biogenic amines was assayed in synthetic media, grape must, and wine. Grape varieties, different types of winemaking, pH, and lactic acid bacteria may be responsible for the differences observed in the biogenic amine concentrations of the wines analyzed.     

以生物胺变化评价冷藏罗非鱼片腐败进程

为了探讨生物胺指标评价鱼肉腐败进程的可能性及其安全范围,应用反相高效液相色谱法测定冷藏罗非鱼片贮藏过程的生物胺变化,用以判断鱼片的腐败进程。结果表明:新鲜鱼片初始单胺和多胺总量较高,二胺总量很低。贮藏过程中单胺和多胺波动变化并呈下降趋势,尸胺、腐胺含量快速增加成为主要生物胺。含有尸胺+腐胺的二胺、生物胺指数、总生物胺等指标与其两者增长趋势相似。相关分析表明尸胺、腐胺、二胺、生物胺指数、总生物胺与贮藏时间、微生物数量和氨基态氮具有高度相关性。经回归分析表明假单胞菌、肠杆菌的增长与尸胺、腐胺具有重要的对应关系。尸胺、腐胺和二胺可简便有效评价鱼片腐败进程;但综合考虑组胺和酪胺的毒性,则生物胺指数(BAI)更适用,冷藏罗非鱼片生物胺指数的初步判别范围为:<20mg/kg,新鲜;20～40mg/kg,可接受;>40mg/kg,腐败。该研究为冷藏罗非鱼片生物胺的限量标准提供数据参考。     

贮藏温度对炝蟹品质的影响

为阐明不同贮藏温度下炝蟹品质的变化和食用安全性,本试验以三疣梭子蟹为材料制作炝蟹,探讨冻藏(-18℃)和冷藏(4℃)条件下炝蟹感官品质、菌落总数(TVC)、挥发性盐基氮(TVB-N)和生物胺的变化规律,并对其生物胺与其他品质指标进行相关性分析。结果表明,贮藏温度对炝蟹感官品质TVC、TVB-N等指标的劣变具有显著的影响。-18℃冻藏和4℃冷藏条件下,炝蟹感官品质分别于第10天和第6个月超出可接受限值;随着贮藏时间的延长,4℃冷藏条件下炝蟹TVC值显著增加,但至贮藏16 d时仍小于5 lg(CFU·g^-1),TVB-N值于贮藏12 d时超出一级鲜度临界值;-18℃冻藏过程中炝蟹TVC值略有增加,但变化不显著,TVB-N值于第6个月超出一级鲜度临界值。4℃冷藏过程中炝蟹中共检测出8种生物胺,而-18℃冻藏过程中共检测出5种生物胺。此外,色胺、腐胺和酪胺是炝蟹贮藏过程中产生的主要生物胺,且与其他品质均具有显著相关性。本研究结果为炝蟹在贮藏过程中的安全性和货架期预测提供了理论依据。     

Gas chromatography-mass spectrometry assessment of amines in Port wine and grape juice after fast chloroformate extraction/derivatization

A simple, reliable, and sensitive gas chromatography-mass spectrometry method for the quantification of volatile and nonvolatile biogenic amines in Port wines and grape juices was developed and evaluated. The method was based on a previously reported two-phase derivatization procedure with isobutyl chloroformate in a toluene medium, which provides a quantitative reaction in 10 min. Following the derivatization step, the excess of reagent was eliminated by treatment with alkaline methanol. The derivatization procedure was performed directly on 1 mL of sample, avoiding any fastidious and time-consuming cleanup extraction steps. The method allows the simultaneous quantification of 22 amines, which can be found in wines: methylamine, dimethylamine, ethylamine, diethylamine, propylamine, isopropylamine, butylamine, isobutylamine, amylamine, isoamylamine, 2-methylbutylamine, hexylamine, pyrrolidine, piperidine, morpholine, 1,3-diaminopropane, putrescine, cadaverine, 1,6-diaminohexane, 2-phenylethylamine, histamine, and tyramine. Because of the fact that histamine and tyramine derivatives are degraded during the isobutyl chloroformate elimination step, the corresponding determination was made after removal of the excess of derivatizing reagent by evaporating an aliquot of the toluene layer obtained after the reaction. The presented method showed excellent analytical characteristics in what linearity, recovery, repeatability, and limit of detections were respected. It was used to assess the concentration of biogenic amines in juice grapes and Tawny and Vintage Port wines with different aging times. On the whole, the total content of amines in Port wines was low. Most of the amines found in wines have their origin in the raw material used for their elaboration, so the Port winemaking process is not prone to the production of this kind of compounds. Total biogenic amine contents have shown a decrease with the aging of both types of Port wines.     

Chemometric studies of vinegars from different raw materials and processes of production

The phenolic composition, aroma compounds, and organic acid content of 83 vinegars have been determined. Multivariate analysis techniques have been used to classify these vinegar samples according to raw material (white wine, red wine, apple, honey, alcohol, balsamic, and malt) and production process (with and without aging in wood). Cluster analysis grouped the samples according to production process. Only apple and balsamic vinegars were separated from wine vinegars. Alcohol, honey, and malt vinegars were grouped with no aged wine vinegars. Linear discriminate analysis allowed a 88% differentiation according to raw material and 100% according to aging in wood. Besides, from the results obtained, a major role of the volatile compounds in the differentiation of the vinegar samples according to their aging period in wood can be seen.     

电子束辐照对真空包装章鱼生物胺的抑制作用研究

为分析电子束辐照处理对真空包装章鱼制品冷藏过程中生物胺形成的抑制作用,以真空包装的新鲜章鱼腕足段为研究对象,分别进行0、0.5、1.0、1.5、2.0和2.5 kGy剂量的电子束辐照处理,于4℃下冷藏63 d,每隔7 d进行感官品评和生物胺含量检测。结果表明,冷藏期间真空包装的章鱼中共检测出精胺、亚精胺、腐胺、尸胺、组胺和酪胺6种生物胺。电子束辐照处理能抑制章鱼冷藏过程中腐胺、尸胺、组胺和酪胺的生成,且抑制作用与辐照剂量成正比,其中对组胺生成的抑制效果最佳,仅在少数样品中被检出。此外,电子束辐照也抑制了章鱼中亚精胺和精胺在冷藏后期的分解。冷藏过程中的腐胺、尸胺和酪胺含量变化与样品感官品质表现出较好的一致性,高感官品质章鱼样品中的腐胺、尸胺和酪胺含量均≤10 mg·kg-1。综合电子束不同剂量辐照对章鱼感官品质和生物胺的抑制作用,1.0 kGy的电子束辐照剂量是真空包装章鱼4℃冷藏保鲜的最适前处理,其能延长高品质新鲜章鱼货架期至35 d。本研究结果为章鱼加工企业生产高品质章鱼产品提供了理论依据。     

Influence of lupin (Lupinus luteus L. cv. 4492 and Lupinus angustifolius L. var. zapaton) and fenugreek (Trigonella foenum-graecum L.) germination on microbial population and biogenic amines

Microbial population and bioactive amine profile and levels of two lupin species (Lupinus luteus L. cv. 4492 and Lupinus angustifolius L. var. zapaton) and fenugreek (Trigonella foenum-graecum L.) seeds as affected by germination were investigated. Microbial population increased considerably mainly in the first stage of germination (2 days), then small changes in bacterial numbers were observed up to 5 days to levels between 7.8 and 8.9 log colony-forming units/g. Microorganisms belonging to the Enterobacteriaceae family were dominant for the legumes tested. Ungerminated legume seeds contained putrescine, cadaverine, histamine, tyramine, spermidine, and spermine. Bioactive amine levels found in fenugreek seeds were between 3- and 4-fold higher than those found in lupin seeds. The highest total amine levels were found in fenugreek seeds [162 mg/kg of dry weight (dw)], followed by L. angustifolius var. zapaton seeds (84 mg/kg of dw) and, finally, L. luteus cv. 4492 (46 mg/kg of dw) seeds. The concentration of individual amines showed a gradual rising trend during the germination period in all tested sprouts, reaching levels >3 times higher than those found in ungerminated seeds. After 5 days of germination, the fenugreek sprouts contained the highest amount of total bioactive amines. Tyramine was the predominant amine in both lupin varieties, whereas cadaverine was the main bioactive amine detected in fenugreek. The results of this work thus indicated that microbial population and biogenic amine levels in the studied lupin and fenugreek sprouts are not a risk for healthy consumers or for individuals with restricted activity of detoxification enzymes.     

Chemical and sensory changes in Mediterranean hake (Merluccius merluccius) under refrigeration (6-8 degrees C) and stored in ice

Fresh Mediterranean hake (Merluccius merluccius var. mediterraneus), a species mainly caught off the shores of Spain, was stored at usual temperatures: in ice (commercial chain) and under refrigeration (home). Sensory and chemical analyses were performed throughout the storage time to determine the changes that took place and evaluate the effect of the storage temperature. Storage in ice resulted in a slight accumulation of volatile and biogenic amines in hake. When it was stored at 6-8 degrees C, a significant production of both trimethylamine and total volatile basic nitrogen (TVB-N) was observed, and biogenic amines were formed. Sensory analysis revealed that hake stored in ice was inedible after 29 days, the figure for refrigerated hake being 20 days. There was a nonsignificant correlation (p > 0.05) between TVB-N values and sensory score in hake stored at 0 degrees C. In all other cases, a significant correlation (p < 0.001) between volatile parameters and sensory analysis was found.     

Development of stable isotope dilution assays for the simultaneous quantitation of biogenic amines and polyamines in foods by LC-MS/MS

Microbial amino acid metabolism may lead to substantial amounts of biogenic amines in either spontaneously fermented or spoiled foods. For products manufactured with starter cultures, it has been suggested that certain strains may produce higher amounts of such amines than others; however, to support efforts of food manufacturers in mitigating amine formation, reliable methods for amine quantitation are needed. Using 10 isotopically labeled biogenic amines as the internal standards, stable isotope dilution assays were developed for the quantitation of 12 biogenic amines and of the 2 polyamines, spermine and spermidine, in one LC-MS/MS run. Application of the method to several foods revealed high concentrations of, for example, tyramine and putrescine in salami and fermented cabbage, whereas histamine was highest in Parmesan cheese and fermented cabbage. On the other hand, ethanolamine was highest in red wine and Parmesan cheese. The results suggest that different amino acid decarboxylases are active in the respective foods depending on the microorganisms present. The polyamine spermine was highest in salami and tuna.     

Content of biogenic amines in a Chardonnay wine obtained through spontaneous and inoculated fermentations

This paper describes the content of biogenic amines in wines obtained from a Chardonnay must inoculated with different strains of Saccharomyces cerevisiae and in a wine fermented with the indigenous yeasts (control wine). The concentrations of nonvolatile amines and phenethylamine in the wines from the inoculated must were superior to those of the control wine. This was probably due to the fact that consumption of the precursor amino acids of these amines, during fermentation, was also greater in the inoculated samples than in the control sample. Furthermore, from the results obtained it may be said that, at least to some extent, the nonvolatile amines were formed by yeasts during fermentation. The concentrations of dimethylamine, ethylamine, and pyrrolidine (volatile amines) were different for the different wines, although they did not reach concentrations sufficiently high to have any effect on the aroma.     

Effects of high hydrostatic pressure treatments on biogenic amine contents in goat cheeses during ripening

The application of high-pressure treatment to accelerate cheese ripening may influence the occurrence and profile of biogenic amines. In some cases, high-pressure treatment could yield high amine content, whereas in others the amine concentrations were comparable with those in nontreated ones. Tyramine was the most affected amine. However, the influence of high pressure depended on the treatment applied. The highest amine concentrations were observed when 50 MPa for 72 h was applied, and in which, the content of tyramine was almost three times higher than in the untreated samples. On the contrary, when 400 MPa for 5 min or 400 MPa for 5 min plus 50 MPa for 72 h were applied contents of this amine were similar or lower than those found in control cheeses showing similar degrees of proteolysis. The higher proteolysis, induced by the pressurization treatments, was not correlated with the higher amine production.     

Study of amine composition of botrytized grape berries

Aliphatic primary and biogenic amines of grape varieties from two vintages were studied. We established that appearance and/or increase of both primary aliphatic and biogenic amines is due to microbiota living in/on grape berries. Aszú, gray rotten grapes infected mainly with Botrytis cinerea, grape berries infected mainly with Penicillium species, and intact grape berries were compared on the basis of amine composition using t-test, analysis of variance, and multivariate statistical analysis (principal component analysis and linear discriminant analysis). The amine composition and amine content of Aszú grapes were significantly different (p < 0.05) from those of intact grapes despite the effect of the vineyards and the vintages. Grape samples coming from the same growing location, intact, Aszú grapes, and grape berries infected mainly with Penicillium species could be separated from each other with multivariate statistical analysis. We distinguished intact, Aszú (noble rotten), and gray rotten berries from each other as well. Evaluating amine values of grape samples independently of the place of origin, the Aszú and green rotten berries could not be differentiated. The varieties and vineyards have affected the amine composition of Aszú grapes, while these effects on intact grapes appeared only slightly.     

Multiple compound quality index for cold-smoked salmon (Salmo salar) developed by multivariate regression of biogenic amines and pH

Production of biogenic amines during chill storage of 12 lots of cold-smoked salmon was studied. These data allowed for a multiple compound quality index to be developed by multivariate regression (partial least square regression). The quality index was based on concentrations of cadaverine, histamine, putrescine, and tyramine and pH and showed good correlation with sensory assessments. Biogenic amines were indicators of spoilage rather than casual agents of spoilage off-flavors. Four different biogenic amine profiles were found at the time of spoilage in cold-smoked salmon. These were the results of differences in the spoilage microflora. Histamine was detected above regulatory limits but below toxic levels. Measurements of salt and dry matter for calculation of water phase salt could be substituted by rapid water activity measurements.     

Changes in amine concentrations during aging of red wine in oak barrels

This investigation studied the evolution of amines in red wines made with Merlot variety, during aging in American oak barrels (Quercus alba) and in French oak barrels (Quercus sessilis) from the Allier and Nevers regions. From the results obtained it was observed that the evolutions of the amines were similar in all three types of oak woods. Histamine and tyramine were produced at the beginning of the aging process, although they were not accumulated in the wines, probably due to their degradation. Putrescine was the most abundant amine in the wines; its concentration increased to an important extent during aging as it did not undergo degradation. The concentration of cadaverine increased slightly at the first stage of aging and, like putrescine, did not degrade at all. The volatile amines showed slight variations during aging, although in no cases were high accumulations observed in the wines. Dimethylamine and isobutylamine were degraded during storage in the barrels.     

降解亚硝酸盐和生物胺乳杆菌筛选及其改善鱼肉香肠品质效果

为了筛选出优良亚硝酸盐和生物胺的降解植物乳杆菌,并研究在鱼肉发酵香肠中的应用。该文主要考察了19 株植物乳杆菌亚硝酸盐和生物胺的降解活性及相关基因的携带情况,从中筛选出优良菌株,并研究其对鱼肉香肠发酵过程中各种亚硝胺、生物胺、腐败微生物的抑制作用和对其他理化指标的影响。试验结果表明,L. plantarum CP3对亚硝酸盐和生物胺的降解能力最强,分别达到95%和77%,PCR结果显示此菌株中含有亚硝酸盐降解（假定蛋白（hypothetical protein,HP）、谷胱甘肽还原酶（glutathione reductase,GR）、一氧化还原酶（Oxidoreductase,ORD））和生物胺降解的相关基因（multicopper oxidases,suf I）,且不含生物胺生成相关基因（hdc,tdc,odc）。GC-MS测定结果显示L. plantarum CP3对发酵香肠中4种亚硝胺-二乙基亚硝胺（N-nitrosodiethylamine,NDEA）,二甲基亚硝胺（N-nitrosodimethylamine,NDMA）,亚硝基吗啉（N-nitrosomorpholine,NMOR）和甲基乙基亚硝胺（N-nitrosomethylamine,NMEA）的生成都具有显著的抑制作用,联合接种L.saki M4可以协同提高亚硝胺的降解能力。并且,与对照组相比,接种L. plantarum CP3+L.saki M4的鱼肉香肠,发酵48 h后腐胺、尸胺和酪胺的含量分别降低了76.83%,93.33%和88%。另外,接种乳酸菌处理组香肠中的肠杆菌、假单胞菌等腐败菌及pH值、硫代巴比妥酸（thiobarbituric acid,TBA）和挥发性盐基氮（total volatile basic nitrogen,TVB-N）的含量显著低于对照组。该研究结果体现了L. plantarum CP3作为香肠等肉制品发酵剂的良好开发应用前景。