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。     

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。     

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。     

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.     

Influence of the freshness grade of raw fish on the formation of volatile and biogenic amines during the manufacture and storage of vinegar-marinated anchovies

Volatile and biogenic amines from three batches of anchovies, marinated in vinegar, were studied. The anchovies had been vacuum-packed and kept in refrigerated storage for 3 months. Trimethylamine and total volatile basic nitrogen levels were very low and constant throughout marinating and storage process: less than 1 and 10 mg/100 g, respectively. Certain amine levels, mainly those of tyramine and serotonin, increased slightly, particularly during storage. However, even the highest recorded levels were much lower than those considered to be hazardous for human consumption. To study the influence of raw material freshness in the amine profile, two laboratory trials using fresh and spoiled anchovies and simulating the industry standard marinating process were carried out. Levels for both volatile and biogenic amines were dependent on raw material quality, proving consistently higher in those deriving from nonfresh fish. Vinegar marinating leads to a decrease in the accumulation of amines in anchovy while their concentration in the vinegar solution increases due to the vinegar effect as solvent extractor.     

Relation of biogenic amines' formation with microbiological and sensory attributes in Lactobacillus-inoculated vacuum-packed rainbow trout (Oncorhynchus mykiss) fillets

The biogenic amine (BA) content of vacuum-packed filleted rainbow trout (Oncorhynchus mykiss) inoculated or not with two different Lactobacillus strains, individually or in combination, was monitored during refrigerated storage for 20 days and related to respective bacteriological and sensory changes occurring during the same period. Eight amines, namely putrescine, cadaverine, tyramine, tryptamine, beta-phenylethylamine, histamine, spermine, and spermidine, were determined, whereas agmatine was not detected in any of the samples. In all cases, BA concentration was higher (P < or = 0.05) in the controls compared to all inoculated treatments, whereas the trend with regard to the bacterial populations (Enterobacteriaceae, pseudomonads, and H2S-producing bacteria) and the off-odor scores was similar. Inoculation with Lactobacillus sakei CECT 4808 showed the best preservative effect among inoculated treatments. Concentrations of putrescine and cadaverine, the main BAs formed, correlated well with both spoilage bacterial counts and off-odor scores and can be useful indicators of shelf life. Spermine and spermidine contents decreased during storage, while levels of the other determined BAs remained below 10 mg/kg even after sensory rejection.     

High-performance liquid chromatographic method for determination of biogenic amines in feedstuffs, complete feeds, and animal tissues

Numerous methods to analyze biogenic amines in biological materials have been described. A versatile and rapid methodology to analyze these compounds in feedstuffs, complete feeds, and animal tissues, however, has not been reported. The current method was developed to address this need. Biogenic amines in feedstuffs, complete animal feeds, and animal tissues were extracted with 10% trichloroacetic acid, reacted with O-phthaladehyde using high-performance liquid chromatographic employing a cation exchange column. Detection limits were 50 pmol/mL for tyramine, histamine, putrescine, and spermine; 40 pmol/mL for cadaverine; and 25 pmol/mL for spermidine. Extraction efficiency of biogenic amines in feedstuffs, duodenum, liver, ileum + jejunum, and whole shrimp and shrimp hepatopancreas ranged between 99-105, 93-135, 80-85, 65-102, 88-98, and 88-97%, respectively. It can be concluded that the current method can be applied to individual feedstuffs, complete feeds, and animal tissues for the rapid and accurate determination of concentration of biogenic amines.     

Quantitative determination of amines in wine by liquid chromatography

A liquid chromatographic (LC) procedure is described for the determination by dansylation of the following 16 kinds of biogenic amines found in wine: monomethylamine (MM), ethylamine (EM), iso- and n-propylamine (Pr), iso- and n-butylamine (Bu), iso- and n-amylamine (Am), pyrrolidine (PY), 2-phenethylamine (PH), tryptamine (TR), putrescine (PU), cadaverine (CA), histamine (HI), tyramine (TY), and spermidine (SP). The amines in white and red wine were applied to a column of Amberlite CG-50 type I resin (Na-form) after the column had been washed with water and eluted with 1N hydrochloric acid. This eluate was evaporated to dryness under reduced pressure and derivatized with dansyl chloride (DNS). LC separations were performed on Finepak SIL C18S and LiChrosorb RP-8 columns with an acetonitrile-water elution gradient. In the survey of commercial wines by this method, most of the samples were found to contain 12 amines, including iso-Am, CA, PU, TY, and others. The highest levels of these amines were 4.84 micrograms PU/mL in red wine, and 5.11 micrograms iso-Am/mL in white wine. The total levels of amines in red wine were comparatively higher than in white wine.     

Chemical, microbiological, and AromaScan evaluation of mahi-mahi fillets under various storage conditions

The quality for mahi-mahi stored at 1.7, 7.2, and 12.8 degrees C for 0, 1, 3, and 5 days was determined using biogenic amine analysis, microbial counts, and sensory evaluation (by a sensory test panel and an AromaScan). Biogenic amines in methanol extracts from mahi-mahi samples were analyzed using capillary electrophoresis (CE) with ultraviolet detection at 210 nm and a gas chromatography (GC) method that can simultaneously determine the contents of putrescine, cadaverine, histamine, spermidine, and spermine within 20 min after pentafluoropropionic anhydride derivatization. A good correlation R2= 0.99) was found between CE and GC methods for detecting histamine in mahi-mahi. Fish quality deteriorated and correlated with increasing microbial numbers. Biogenic amines may be useful indicators for mahi-mahi quality and safety. AromaScan was able to correlate quality changes for mahi-mahi in microbiological and sensory analyses.     

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.     

High-performance liquid chromatographic determination of biogenic amines in poultry carcasses

Biogenic amines, produced by bacterial decarboxylation of amino acids, have been associated with toxicological symptoms in broilers fed various poultry byproducts. A reversed-phase high-performance liquid chromatographic method is described for the quantitation of eight biogenic amines (tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine, and spermine) in chicken carcasses. Amines were extracted with perchloric acid, derivatized with dansyl chloride, separated using gradient elution (methanol and water), and detected by fluorescence. Benzylamine was used as the internal standard. Linearity, repeatability, and recovery of the method were evaluated. The method was linear for all of the amines studied at concentrations ranging from 0.05 to 25 microg/mL. Average recoveries ranged from 92.6% to 96.8% for all amines except for histamine, which was 74.6%.     

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.     

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.     

Influence of cultivar and germination on bioactive amines in soybeans (Glycine max L. Merril)

The levels of amines in soybeans as affected by cultivar in two consecutive years and by germination were investigated. Spermidine, spermine, putrescine, agmatine, and cadaverine were detected, whereas tyramine, histamine, tryptamine, serotonine, and phenylethylamine were not. Spermidine was the predominant amine followed by spermine. High concentrations of these amines confirmed soybean as a rich source. Cadaverine was confirmed to be inherent to soybean. The percent contribution of spermidine and spermine to total levels was not affected by cultivar in either years. However, amine levels were affected by cultivars in different ways in the consecutive years. Cadaverine was affected more by the cultivar, whereas spermidine, spermine, and agmatine were affected by harvest year. During germination the levels of amines from soybean increased significantly, except for agmatine. Spermidine and spermine accumulated in the cotyledon, whereas cadaverine and putrescine accumulated in the radicle and hypocotyl.     

HPLC method for analysis of free amino acids in fish using o-phthaldialdehyde precolumn derivatization

Precolumn derivatization applying o-phthaldialdehyde (OPA) was used to analyze free lysine, histidine, and ornithine, precursors of the respective biogenic amines cadaverine, histamine, and putrescine, which are considered indicators of fish quality and safety. This method uses 75% methanol to eliminate the use of strong acids as the extraction solution. Each analysis took 35 min, was reproducible, and allowed separation of primary amino acids in fish samples. A binary solvent delivery system coupled with a fluorescence detector and an Ultrasphere ODS column were utilized for HPLC separation. Linearity of the calibration curves was very good (r(2) = 0.99) for the amino acids of interest. Minimum concentrations of detection were 40 pmol/mL for histidine and lysine and 70 pmol/mL for ornithine. Average recoveries were 72% for lysine, 93% for histidine, and 98% for ornithine. This method used solvent gradient elution to study the levels of these analytes in mahi-mahi, bigeye tuna, and flounder.     

Biogenic amines and polyamines in milks and cheeses by ion-pair high performance liquid chromatography

The proposed chromatographic method provides a complete resolution of twelve amines in a single run in milks and unripened cheeses, avoiding the losses of resolution linked to fluctuations in working temperature. We also propose an alternative chromatographic gradient, which can be useful for samples that have undergone long ripening periods, like ripened cheeses. According to the results of the reliability study, the method described was precise, accurate, and sensitive. The method was applied to several samples of milks and cheeses and the results showed that the biogenic amine profiles varied greatly, not only between different types of samples but also among the samples from the same kind of products. In unripened cheeses, milks, and yogurts, spermidine and spermine were the prevailing amines, but in ripened cheeses the major amine was tyramine, followed by putrescine and cadaverine.     

Simultaneous determination of four biogenic and three volatile amines in anchovy by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry

A new method has been developed and validated for the simultaneous determination of four biogenic (putrescine, cadaverine, histamine, and tyramine) and three volatile amines (trimethylamine, triethylamine, and tripropylamine) in anchovy. Separation and determination of the selected compounds were carried out by ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS), using an electrospray ionization source (ESI) in positive ion mode. Chromatographic separation was carried out using an aqueous solution of formic acid (0.1%) and methanol as mobile phase in gradient mode. The method was validated, and mean recoveries were evaluated at three concentration levels (75, 150, and 250 μg/kg), ranging from 70 to 110% at the three levels assayed. Intra- and interday precision, expressed as relative standard deviation (RSD), were lower than 15% and 20%, respectively. Limits of quantitation (LOQs) were 25 μg/kg for all cases, except for that of TMA, which was set at 60 μg/kg. The developed procedure was applied to determine the target compounds in anchovy samples stored during 7 days at 4 °C, observing the increasing in the concentration of these compounds at longer storage time.     

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.