Production of Biogenic Amines and Their Potential Use as Quality Control Indices for Hake (Merluccius merluccius, L.) Stored in Ice

ABSTRACT: The production of tyramine, histamine, putrescine, cadaverine, agmatine, and spermidine was studied for use as quality indices in muscle of gutted hake stored in ice for 25 d. The changes in biogenic amine concentrations were correlated with trimethylamine nitrogen levels. All concentrations except for spermidine, increased progressively throughout storage. During storage, cadaverine reached 72.14 mg/kg while there were no significant increases in histamine and tyramine. The levels of free amino acids, which may be metabolized into biogenic amines, were not correlated to respective biogenic amines levels. The production of cadaverine and agmatine occurred prior to spoilage and these biogenic amines may indicate freshness of hake stored in ice.     

BIOGENIC AMINES IN FINNISH DRY SAUSAGES

A study was conducted to determine the biogenic amine levels of Finnish dry sausages and to compare the results with those of other surveys of different types of fermented sausages. Sausages with high amine concentration were analyzed microbiologically in order to identify amine-producing microorganisms. Tyramine was the most common amine: the mean of 68 samples was 82 mg/kg and the mode 110 mg/kg. Other vasoactive amines found were histamine (<1–200 mg/kg), phenylethylamine (<1–48 mg/kg) and tryptamine (<10–91 mg/kg). Increased amounts of the spoilage indicator amines putrescine and cadaverine were found from most of the samples containing vasoactive amines. The formation of amines can be linked with problem(s) in hygienic conditions of raw materials and manufacturing practices and it is thus proposed that the sum of tyramine, histamine, putrescine and cadaverine could serve as an indicator of quality in fermented meat products.     

Effect of controlled atmospheres enriched with O2 in formation of biogenic amines in chilled hake (Merluccius merluccius L.)

Changes in biogenic amines (tyramine, histamine, putrescine, cadaverine, agmatine and spermidine) contents during the storage of chilled hake in a controlled atmosphere with four gas mixtures of different CO₂ and O₂ concentrations, (CO₂/O₂/N₂ at 60/15/25, 40/40/20, 60/40/0 and 40/60/0 respectively) were studied. Biogenic amines were determined by high-pressure liquid chromatography with post-column derivatization and ortho-phtaldehyde. Histamine levels, potentially the most toxic amine in hake, were very low during storage. The amine in hake with the highest level was cadaverine. Spermidine and putrescine had very high levels at the beginning of the storage. However, agmatine and tyramine were not detected in the source raw material. An increase in biogenic amines was observed in the control lot from day 12 of storage except for agmatine, where levels began to increase earlier. Cadaverine and agmatine could be used as control indices due to their behaviour during the storage period studied. The varying O₂ and CO₂ concentrations in the different atmospheres did not seem to affect the production of spermidine. The high levels of CO₂ in thelot in M1 atmosphere were not sufficient to inhibit the production of biogenic amines. However, the high O₂ concentration had an inhibiting effect on the production of biogenic amines. Therefore, the formation of biogenic amines was inhibited by controlled atmospheres with oxygen-enriched binary gas mixtures.     

Biogenic Amine Content of Beers Consumed in Turkey and Influence of Storage Conditions on Biogenic Amine Formation

The biogenic amine content of beers available in Turkey, both domestic production and imported products, was investigated. A total of 17 domestic and 13 imported beers were evaluated in terms of 8 different biogenic amines (histamine, tyramine, spermine, spermidine, 2‐phenylethylamine, putrescine, tryptamine and cadaverine). HPLC methodology with pre‐column derivatization and photodiode array detection after derivatization with dansyl chloride was used for quantification. In addition, the evolution of important amines such as histamine, tyramine, putrescine and cadaverine were investigated during different storage conditions by response surface methodology. The results indicated that both storage temperature and storage time were important factors affecting biogenic amine content. Histamine and putrescine increased during storage, but then decreased after reaching a maximum level after six weeks. With the biogenic amines tyramine and cadaverine, the amounts increased during the entire storage period. At higher storage temperatures, the formation of biogenic amines increased.     

白鱼腐败菌产生物胺能力分析

为了探讨白鱼（Anabarilius）腐败菌产生物胺的能力,釆用聚合酶链式反应技术检测腐败菌的组氨酸、赖氨酸、鸟氨酸和酪氨酸的脱羧酶基因,应用反相高效液相色谱技术测定接种腐败菌培养基和无菌白鱼鱼块中生物胺含量,并对主要腐败菌的生物胺产量因子进行分析。结果表明：8 种腐败菌均含有至少1 种氨基酸脱羧酶基因;接种不同腐败菌的液体培养基和无菌白鱼鱼块均检测到多种生物胺,其中Morganella sp.wf-1和Morganella sp.wf-2具有很强的产腐胺、尸胺和组胺能力,使得接种这两种腐败菌的无菌白鱼鱼块中腐胺、尸胺和组胺含量、单胺、二胺、生物胺指数和总生物胺含量等指标在腐败过程中均显著高于对照组和接种其他腐败菌的无菌白鱼鱼块。同时,Morganella sp.wf-1和Morganella sp.wf-2的腐胺、尸胺、组胺、单胺、二胺、生物胺指数和总生物胺的产量因子均高于其他腐败菌。结果表明,Morganella sp.wf-1和Morganella sp.wf-2是白鱼腐败过程中产生物胺的主要腐败菌。     

利用高效液相色谱法测定不同发酵时长郫县豆瓣酱中的8种生物胺

本研究基于高效液相色谱(HPLC)技术,测定了不同发酵时长(1~96个月)郫县豆瓣酱中色胺、β-苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺8种生物胺的含量。结果显示,HPLC分析中,8种生物胺的检出限在0.17~0.35 mg/L之间,定量限在0.55~0.92 mg/L之间,各生物胺标品在1~60 mg/L的质量浓度范围内线性关系良好,可被准确定量;各生物胺回收率在73.10%~104.13%之间,说明本方法具有较高的重复性和灵敏度。腐胺、尸胺、酪胺和组胺是郫县豆瓣酱中的主要生物胺;随着发酵时间的延长,腐胺含量快速上升,尸胺含量在发酵前54个月迅速上升,之后明显下降,酪胺和组胺在发酵54个月后含量有所增加;其他生物胺含量较低,且发酵期间变化不明显;郫县豆瓣酱中总生物胺含量随发酵时间的延长不断积累,从(37.86±6.69) mg/kg增加到(155.09±8.53) mg/kg,其含量在发酵前54个月快速上升,之后趋于稳定。目前食品中的多数生物胺尚没有限量标准,但参考现有的毒理学数据来看,本研究中郫县豆瓣酱的生物胺含量水平相对安全。     

发酵剂对牦牛乳硬质干酪成熟过程中生物胺的影响

乳酸菌产生物胺的能力具有菌株特异性,因此,为了探究不同种类发酵剂对牦牛乳硬质干酪中生物胺形成的影响,该试验利用高效液相色谱对3种不同发酵剂制作的硬质干酪成熟过程中生物胺进行了测定和分析。结果表明,嗜热和嗜温发酵剂牦牛乳硬质干酪中检测出2-苯乙胺、腐胺、尸胺、组胺和酪胺,混合发酵剂干酪中检测出腐胺、2-苯乙胺、尸胺和酪胺。各生物胺之间呈现正相关性。3种不同发酵剂干酪在1～6个月成熟过程中,其各生物胺整体呈现增加趋势,嗜热、嗜温和混合发酵剂干酪中总生物胺最高含量分别为(448.3±9.6)、(456.8±58.4)、(293±24.5)mg/kg。组胺和酪胺是2种毒性相对高的生物胺,嗜热发酵剂干酪中组胺和嗜温发酵剂干酪中酪胺最高,其最高含量分别为(20.8±7.9)、(92.9±6.7)mg/kg,混合发酵干酪中未检测出组胺,酪胺含量次之,3种不同发酵剂干酪中组胺、酪胺含量均低于推荐安全剂量50 mg/kg和100 mg/kg。这为合理选择发酵剂和控制干酪中生物胺形成提供了依据。     

我国市售不同类型及品牌腐乳中生物胺含量的分析与比较

为了揭示我国商业腐乳中生物胺存在形式和水平,采用丹磺酰氯柱前衍生结合高效液相色谱法（HPLC）对我国市售的20个品牌白方腐乳和20个品牌红方腐乳中的生物胺含量进行了分析。结果表明,我国市售的腐乳中存在的主要生物胺为酪胺、组胺、腐胺、色胺、β-苯乙胺和尸胺,其平均含量分别为73.7 mg/kg、43.7 mg/kg、38.5 mg/kg、27.7 mg/kg、5.4 mg/kg和5.4 mg/kg;每种生物胺在白方腐乳中的含量均显著高于红方腐乳（P＜0.05）,前者的酪胺、组胺、腐胺、色胺、β-苯乙胺和尸胺的平均含量分别为后者的1.6、1.9、3.4、3.3、2.3和3.0倍;且每种生物胺的水平在同一类型不同品牌腐乳之间均呈现高度差异化。     

Biogenic amines in fish, fish products and shellfish: a review

Fish, cephalopods and shellfish provide a healthy source of high-quality proteins, essential vitamins, minerals and polyunsaturated fatty acids. The beneficial effects of fish consumption on human health such as protection against coronary heart disease and certain cancer may be offset by fish decomposition and the formation of chemical contaminants such as biogenic amines. There are several toxicological effects of biogenic amines on humans, especially histamine. It is the causative agent of histamine or scombroid fish poisoning which is a significant public health problem. In individuals with diminished histamine detoxification, ingestion of even a low or moderate histamine- or tyramine-containing fish may lead to food intolerance. Biogenic amines such as putrescine, tyramine and cadaverine can potentiate histamine toxicity. Furthermore, dietary polyamine intake should be minimised in some cancer patients. Besides their potential toxicity, biogenic amines are used for the evaluation of hygienic quality of different marine and freshwater species. Spoilage pattern and biogenic amine formation are species specific. Histamine has been traditionally used as an indicator of the quality of histidine-rich fish (dark-muscle fish). On the other hand, putrescine and cadaverine are the most objective indicators of quality of histidine-poor fish (white-muscle fish), shellfish and fermented seafood products.     

Biogenic amines in fish,fish products and shellfish: a review

Fish, cephalopods and shellfish provide a healthy source of high-quality proteins, essential vitamins, minerals and polyunsaturated fatty acids. The beneficial effects of fish consumption on human health such as protection against coronary heart disease and certain cancer may be offset by fish decomposition and the formation of chemical contaminants such as biogenic amines. There are several toxicological effects of biogenic amines on humans, especially histamine. It is the causative agent of histamine or scombroid fish poisoning which is a significant public health problem. In individuals with diminished histamine detoxification, ingestion of even a low or moderate histamine- or tyramine-containing fish may lead to food intolerance. Biogenic amines such as putrescine, tyramine and cadaverine can potentiate histamine toxicity. Furthermore, dietary polyamine intake should be minimised in some cancer patients. Besides their potential toxicity, biogenic amines are used for the evaluation of hygienic quality of different marine and freshwater species. Spoilage pattern and biogenic amine formation are species specific. Histamine has been traditionally used as an indicator of the quality of histidine-rich fish (dark-muscle fish). On the other hand, putrescine and cadaverine are the most objective indicators of quality of histidine-poor fish (white-muscle fish), shellfish and fermented seafood products.     

Formation of biogenic amines throughout the industrial manufacture of red wine

Changes in biogenic amines (histamine, methylamine, ethylamine, tyramine, phenylethylamine, putrescine, and cadaverine) were monitored during the industrial manufacture of 55 batches of red wine. The origin of these amines in relation to must, alcoholic fermentation, malolactic fermentation, sulfur dioxide addition, and wine aging and the interactions between amines and their corresponding amino acids and pH were statistically evaluated in samples from the same batches throughout the elaboration process. Some amines can be produced in the grape or the musts (e.g., putrescine, cadaverine, and phenylethylamine) or can be formed by yeast during alcoholic fermentation (e.g., ethylamine and phenylethylamine), although quantitatively only very low concentrations are reached in these stages (less than 3 mg/liter). Malolactic fermentation was the main mechanism of biogenic amine formation, especially of histamine, tyramine, and putrescine. During this stage, the increase in these amines was accompanied by a significant decline in their amino acid precursors. Significant correlations between biogenic amine formation and the disappearance of their corresponding amino acids were observed, which clearly supports the hypothesis that malolactic bacteria are responsible for accumulation of these amines in wines. No increase in the concentration of biogenic amines was observed after SO2 addition and during wine aging, indicating that sulfur dioxide prevents amine formation in subsequent stages.     

Determination of Biogenic Amines in Sichuan-Style Spontaneously Fermented Sausages

The qualitative and quantitative profile of biogenic amines in 30 samples of Sichuan-style spontaneously fermented sausages sold in Sichuan retail markets was studied by HPLC. Tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, and spermidine were analyzed by UV detection after pre-column derivatization with dansyl chloride. There were wide ranges of different biogenic amines in the samples, except spermidine. The results showed that the histamine was the major biogenic amines, followed by tyramine and cadaverine, which have a mean value of 196.06 mg/kg (ranging from 114.33 to 327.35 mg/kg), 164.67 mg/kg (ranging from 88.84 to 285.89 mg/kg), and 141.65 mg/kg (ranging from 19.14 to 376.46 mg/kg), respectively. The toxicity limit of histamine is 100 mg/kg, where all of the samples exceeded this limit. None of the samples of total biogenic amines content exceeded 1000 mg/kg. The results showed that the amounts of biogenic amines in Sichuan-style spontaneously fermented sausages sold in Sichuan retail markets may pose a potential health risk for sensitive individuals. This suggested that it is necessary to improve the production techniques and the raw materials’ hygienic quality, to ensure the quality and safety of this fermented sausages with respect to their biogenic amine content.     

Biogenic amine content during the manufacture of dry-cured lacón, a Spanish traditional meat product: Effect of some additives

The content of nine biogenic amines (agmatine, tryptamine, 2-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine) was determined throughout the manufacture of dry-cured lacón, a traditional dry-salted and ripened meat product made in the north-west of Spain from the fore leg of the pig following a similar process to that of dry-cured ham. The effect of the use of additives (glucose, sodium nitrite, sodium nitrate, sodium ascorbate and sodium citrate) on the biogenic amine content during manufacture was also studied. Tryptamine and spermine were the main biogenic amines in fresh meat, while tryptamine and cadaverine were the most abundant at the end of the manufacturing process. During ripening the total amine content increased significantly (P<0.05) in the batches made both without and with additives. The use of additives significantly (P<0.05) increased the total amine content and the content of tryptamine, tyramine and histamine. The total biogenic amine content at the end of the manufacturing process was low as expected for a product in which there is little active microbial metabolism during manufacture.     

发酵香肠中生物胺生成量的菌株效应分析

本实验对乳杆菌(米酒乳杆菌和戊糖乳杆菌)和片球菌(戊糖片球菌和小片球菌)在发酵香肠中生物胺的生成量进行了分析。结果表明,供试乳杆菌和片球菌均能产生不等量的组胺、尸胺、腐胺、酪胺、亚精胺和精胺,但都不产生色胺,产品中的生物胺生成量表现出明显的菌株效应。采用复配菌种(片球菌+葡萄球菌)发酵可以明显降低香肠中组胺、尸胺、腐胺和酪胺以及总生物胺的含量。     

温度和姜精油对金枪鱼品质影响及生物胺相关性

将分别贮藏于不同温度(0、4、15℃)条件下的金枪鱼样品,以及4℃条件下用姜精油作为生物抗氧化剂处理过的样品作为研究对象,定期测定金枪鱼生物胺(腐胺Put、尸胺Cad、组胺His、酪胺Tyr)、反式尿刊酸的含量、挥发性盐基氮(total volatile basic nitrogen,TVB-N)值、K值和菌落总数,研究温度和姜精油对金枪鱼生物胺含量的影响及组胺与反式尿刊酸的关系。结果表明,温度对金枪鱼生物胺生成有较大影响,0℃条件下生物胺含量显著低于其他贮藏温度,到第6天时组胺、酪胺、腐胺和尸胺含量分别为28.25、5.47、14.84 mg/kg和17.05 mg/kg,而此时15℃条件下金枪鱼组胺、酪胺、腐胺和尸胺含量分别为135.4、14.63、29.49 mg/kg和41.55 mg/kg;生物胺中组胺含量最高,而酪胺含量始终处于低水平状态,即使在15℃条件下到第7天时其含量仍为16.24 mg/kg;结果同时表明,生物抗氧化剂对生物胺的产生和微生物生长有一定的抑制作用,并能有效地延缓蛋白质和ATP降解。相关分析和回归分析表明,组胺、酪胺、尸胺、与TVB-N值、K值和菌落总数高度相关,其相关系数r在0.804~0.981之间,且生物胺和菌落总数、TVB-N值之间存在重要对应关系,反式尿刊酸对组胺的产生影响不大,两者之间相关性较弱,在4℃时其相关系数r为0.630。     

In vitro synthesis of biogenic amines by Brochothrix thermosphacta isolates from meat and meat products and the influence of other microorganisms

Twenty Brochothrix thermosphacta strains tested for biogenic amines (BAs) production, formed histamine (6.6-16.2 mg/kg) and tyramine (18.7-35.4 mg/kg) but neither putrescine nor cadaverine. Six of the twenty strains were also investigated in respect of their influence on the synthesis of BAs by Escherichia coli, Pseudomonas sp., Proteus mirabilis and Lactobacillus sakei. In pure culture Escherichia coli produced all of the studied amines (histamine, tyramine, putrescine and cadaverine) with a total concentration of 167.7 mg/kg, P. mirabilis produced a total of 56.7 mg/kg histamine, tyramine and cadaverine, while Lactobacillus sakei and Pseudomonas sp. produced histamine and tyramine, totaling 37.9 and 35.2 mg/kg respectively. All B. thermosphacta promoted cadaverine production by Escherichia coli which increased by 12-68%, and some of them contributed to the appearance of this amine among the metabolites of Pseudomonas. The presence of B. thermosphacta decreased the potential ability of P. mirabilis to produce BAs.     

CHANGES IN BIOGENIC AMINE CONTENTS AND SENSORY QUALITY OF SARDINE (SARDINA PILCHARDUS) STORED AT 4C AND 20C

Biogenic amines (tyramine, putrescine, cadaverine, histamine, spermidine and spermine) in sardine during the storage at two temperatures (4C and 20C) were investigated. Changes in sensory scores were also recorded. Significant differences between storage times were found. The levels of tyramine, putrescine, cadaverine and histamine increased as decomposition of sardine progressed at both 4C and 20C. Spermidine and spermine did not change throughout storage. Putrescine was the main biogenic amine formed, followed by tyramine, cadaverine and histamine. Histamine remained lower than the toxic level of 100 mg/kg in the samples stored at 4C whereas it exceeded the toxic level after 16 h at 20C. The increase in histamine concentration was lower than tyramine and putrescine at both temperatures. Sensory scores gradually decreased during the storage.     

豆豉中生物胺含量的动态变化研究

采用高效液相色谱法对豆豉中生物胺含量进行动态检测。结果表明,纯种强化发酵和自然发酵豆豉样品中均检测出色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺。纯种强化发酵条件下生物胺总含量在前、后发酵时期都呈先上升后下降的趋势,而自然发酵条件下均呈缓慢上升趋势,且前发酵总生物胺含量高于后发酵时期。两种发酵条件下,酪胺和亚精胺为主要生物胺,苯乙胺存在纯种发酵中。纯种强化发酵豆豉总生物胺含量高于自然发酵,酪胺和苯乙胺的含量分别高出2倍和4倍,与生产环境中粪肠球菌的污染关联性较大。豆豉发酵过程和成品中总生物胺含量均＜250 mg/kg,短期内不存在食品安全问题。     

Biogenic amine content of red Spanish wines: comparison of a direct ELISA and an HPLC method for the determination of histamine in wines

Biogenic amines were determined in 61 commercial red Spanish wines from various winemaking areas and elaborated with different vinification/ageing procedures. Biogenic amines (histamine, methylamine, ethylamine, tyramine, phenylethylamine, putrescine and cadaverine) analysis was carried out by RP-HPLC with o-phthaldialdehyde precolumn derivatization and fluorescence detection. Overall, histamine and putrescine were the most prevalent amines, being present in 75% and 71%, respectively, of the wines, followed by tyramine (56% of the wines). There were no significant differences between biogenic amine levels and ageing characteristics of the wines tested. Moreover, it was found that amines suspected to cause toxicological effects (histamine, tyramine and phenylethylamine) are no cause for concern in these Spanish wines as they are present in amounts well below the limit considered as physiological. A commercial competitive direct ELISA immunoassay method specific for the detection of histamine has also tested on these wines and the results were compared with the RP-HPLC method. Both methods showed a good agreement for histamine analysis of wines.     

Volatile and Nonvolatile Amines in Mediterranean Hake as Function of their Storage Temperature

ABSTRACT: The main amines in frozen and fresh hake (Merluccius merluccius) are the natural polyamines, spermidine. The levels of volatile and nonvolatile amines during storage of hake were strogly dependent on temperature. In samples stored at -20° only changes in dimethylamine and agmatine were found, whereas major changes were observed in samples stored at higher temperatures. Cadaverine was the major amine formed followed by histamine, putrescine and tyramine. The maximum levels of biogenic amines were lower than those reported for other fish species. Trimethylamine and biogenic amines related to hake spollage showed levels higher in fresh than in frozen samples, whereas levels of natural polyamines were not statistically different.