传统自然发酵四川香肠加工贮藏过程中生物胺含量变化

采用高效液相色谱法对传统自然发酵四川香肠加工贮藏过程中生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、酪胺和亚精胺)含量变化进行测定。结果表明:传统自然发酵四川香肠原料肉中只检测出了色胺、苯乙胺和亚精胺,贮藏120天检测出了7种生物胺,除亚精胺外,6种生物胺(色胺、苯乙胺、腐胺、尸胺、组胺和酪胺)的含量在加工过程中均显著增加(P0.01)。传统自然发酵四川香肠中以酪胺、组胺、尸胺和腐胺为主,含量分别达到153.06、127.90、132.46和92.83 mg/kg,4种生物胺占生物胺总量的89.66%。传统自然发酵四川香肠中生物胺(色胺、苯乙胺、组胺和酪胺)含量均低于20 mg/100 g。生物胺总量在加工贮藏过程中变化显著(P0.01),总量低于1 000 mg/kg。     

高效液相色谱法同时测定香肠中6种生物胺

采用丹磺酰氯为衍生试剂,建立反相高效液相色谱法同时测定香肠中腐胺、尸胺、酪胺、组胺、亚精胺和精胺6种生物胺的方法.色谱条件:C18色谱柱分离,以甲醇和0.1%乙酸溶液为流动相梯度洗脱,流速0.8mL/min,二极管阵列检测器,检测波长254 nm.以1,7-二氨基庚烷为内标,在设定的试验条件下,6种生物胺实现了良好的分离,并呈良好的线性相关性(r>0.999).方法的检测限:腐胺、亚精胺、尸胺和酪胺均为0.8μg/mL,组胺和精胺均为4 μg/mL.本检测方法具有良好的重现性和回收率.对发酵香肠样品中的生物胺含量进行了分析,其中部分香肠中含有一定量的生物胺,最高可达1.09mg/g.     

市售四川香肠中生物胺含量比较分析

利用高效液相色谱,以丹磺酰氯为衍生试剂,分析了四川地区30种市售四川香肠中色胺、苯乙胺、腐胺、尸胺、组胺、酪胺和亚精胺的含量。结果表明:30种市售四川香肠中7种生物胺的检出率均为100%,其中组胺含量最高,平均为182.69 mg/kg;酪胺、尸胺和腐胺次之,平均含量分别为176.93、163.40和60.88 mg/kg;亚精胺平均含量最低,为7.68 mg/kg。30种四川香肠中生物胺的总含量为335.76~1 267.13 mg/kg,1号样品的生物胺总量最高,1、2、5、13、14、15和16号样品中尸胺、组胺和酪胺含量均超过100 mg/kg,存在一定的安全隐患。     

混合发酵剂对川味香肠发酵成熟过程中生物胺含量变化的影响

以植物乳杆菌、戊糖片球菌和葡萄球菌作为发酵剂接种至川味香肠,采用高效液相色谱法测定川味香肠发酵成熟过程中生物胺含量的变化。结果表明:成熟结束时,除苯乙胺外,A组(接种组)中的色胺、腐胺、尸胺、组胺、酪胺和亚精胺的含量显著低于B组(自然发酵组)(P0.01),说明混合发酵剂能显著抑制川味香肠发酵成熟过程中生物胺的形成(P0.01)。A组香肠在发酵成熟过程中除酪胺含量最高达到103.05 mg/kg外,其余生物胺含量均低于100 mg/kg;B组香肠在发酵成熟过程中腐胺、尸胺、组胺和酪胺含量超过100 mg/kg,因此接种混合发酵剂可以更好地保障川味香肠的安全性。     

四川区域自然发酵香肠及人工接种牦牛肉发酵香肠中生物胺含量的研究

以四川省10 区域自然发酵香肠及人工接种发酵剂的牦牛肉香肠共21 个样品为研究对象,测定发酵香肠中的生物胺含量。结果发现,21 个发酵香肠样品中均检测到酪胺、亚精胺、精胺、尸胺、腐胺、色胺及组胺,β-苯乙胺均未检出;生物胺总量在57.34～411.12 mg/kg,除采自凉山州西昌的自然发酵牦牛肉香肠和广安的自然发酵猪肉香肠中酪胺含量超过了美国食品及药品管理局（Food and Drug Administration,FDA）规定标准（酪胺含量≤100 mg/kg）,其余均不存在生物胺安全问题。     

高效液相色谱法同时测定酱油中的8种生物胺

本文建立了高效液相色谱法同时检测酱油中色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺和精胺的方法。采用5%三氯乙酸溶液为样品提取溶剂,提取液经丹璜酰氯柱前衍生30 min,最终HPLC进行定性和定量分析。本方法中8种生物胺的线性范围为1.0~50μg/mL,相关系数R2大于0.99,检出限(S/N=3)为0.075~0.3μg/mL。在添加水平为1.00和5.00μg/mL时,样品的平均回收率在83%~111%之间,相对标准偏差为0.43%~19.0%。本方法具有线性范围广,灵敏度和准确度高等优点,适用于酱油中生物胺的检测。采用本方法对市售10种酱油进行检测,总生物胺含量的范围为50.82~1898.17μg/mL,其中酪胺、腐胺和苯乙胺是酱油样品中含量最多的生物胺。     

国产葡萄酒中生物胺组成与含量分析

通过对国产葡萄酒产品中生物胺组成和含量的检测分析,了解国产葡萄酒中生物胺含量现状。采用GB/T 5009.208—2008方法,对国内市售的250个国产葡萄酒样品中8种生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、精胺和亚精胺)含量进行检测。结果显示,国产葡萄酒样品中8种生物胺含量范围分别为色胺0~32.78 mg/L,苯乙胺0~48.20 mg/L,腐胺0~46.60 mg/L,尸胺0~9.38 mg/L,组胺0~14.37 mg/L,酪胺0~14.60 mg/L,精胺0~12.73 mg/L,亚精胺0~0.50mg/L;葡萄酒样品中检出率最高和最低的生物胺分别为腐胺和精胺,检出率分别为88.4%和50.0%,组胺检出率为52.8%;98.8%的葡萄酒样品中组胺含量低于10 mg/L,98.0%的葡萄酒样品中酪胺含量低于10 mg/L。同其他种类葡萄酒相比,干红葡萄酒样品中生物胺含量相对较高,但与欧洲葡萄酒传统酿造国家的同类产品含量水平一致。国产葡萄酒中生物胺含量水平符合食品安全要求,具有较好的饮用安全性。     

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。     

液质联用法测定黄酒中生物胺含量

采用高效液相色谱-三重四级杆质谱串联法(HPLC-MS/MS)技术,建立检测黄酒中组胺、腐胺、尸胺、精胺、亚精胺、2-苯乙胺、酪胺、色胺共8种生物胺的分析方法。结果表明,在液质联用法对黄酒中生物胺的检测限方面,组胺、尸胺、精胺、亚精胺为0.05μg/kg,腐胺1.0μg/kg,酪胺、色胺为1.5μg/kg,2-苯乙胺为2.0μg/kg。线性范围为0.05~15.0μg/kg(R0.999),回收率在93%~99%之间,相对标准偏差(RSD)在0.11%~0.29%之间。液质联用法可对黄酒中生物胺进行准确的定性和定量,且样品不需衍生化处理,提高了大批量样品检测前处理的效率,同时解决了检测中出现的假阳性问题,适合黄酒中生物胺的低残留检测要求。     

混合菌种发酵生产低酸度川味香肠加工过程中生物胺含量的变化

接种混合发酵剂并在控温控湿条件下生产低酸度川味香肠,对加工过程中生物胺(色胺、苯乙胺、腐胺、尸胺、组胺、酪胺和亚精胺)含量变化进行研究。结果表明:干燥结束时,低酸度川味香肠中除亚精胺以外,其他生物胺含量均低于人工控温控湿不接种组(B组)、自然条件接种组(C组)、自然条件不接种组(D组),其中色胺、苯乙胺、腐胺、组胺和酪胺的含量均显著低于未接种组(P0.01),说明混合发酵剂在适宜的温湿度条件下能明显抑制低酸度川味香肠中色胺、苯乙胺、腐胺、组胺和酪胺积累(P0.01);混合发酵剂对低酸度川味香肠中尸胺的抑制作用不显著(P0.05)。     

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.     

Changes in biogenic amine and polyamine contents in slightly fermented sausages manufactured with and without sugar

The effect of sugar omission on biogenic amine contents of slightly fermented sausages during ripening and storage was evaluated. Two batches of spontaneously fermented sausages were produced with and without sugars in two different trials at pilot-plant scale. Ripened sausages were stored at 4 and 19°C for a further 20 days. Tyramine and cadaverine were the main amines formed during ripening, their contents being significantly higher in batches without sugar. High counts of LAB and Enterobacteriaceae could be associated with the production of tyramine and cadaverine, respectively. The occurrence of putrescine depended on the trial and batch. Sausages without sugar contained more putrescine than those with sugar in trial 1, but this was not repeated in trial 2, in which a high production of agmatine occurred. Tryptamine and phenylethylamine were only detected in the later stages of ripening, their contents also being higher in sausages without sugar. Biogenic amine contents generally rose during storage at 19°C, the increase being especially important for cadaverine and tyramine in sausages without sugar. Levels of spermidine and spermine remained constant during ripening and decreased slightly during storage. Sugar omission is not recommended because it might increase biogenic amine accumulation during the manufacture and storage of slightly fermented sausages.     

熏马肠成熟过程中产生物胺氧化酶菌对生物胺作用的研究

该研究以从新疆熏马肠中筛选出来的6株产生物胺氧化酶菌株(鼠李糖乳杆菌、枯草芽孢杆、腐生葡萄球菌、木糖葡萄球菌、戊糖片球菌、植物乳杆菌)作为发酵剂加入熏马肠中,采用PCR-DGGE技术监测菌落动态变化,高效液相色谱法检测熏马肠中生物胺(色胺、腐胺、苯乙胺、组胺、酪胺、尸胺、精胺、亚精胺)含量变化。结果表明,在熏马肠成熟过程中发酵剂对生物胺有一定的抑制作用。其中以腐生葡萄球菌对色胺、腐生葡萄球菌对苯乙胺、木糖葡萄球菌对尸胺、鼠李糖乳杆菌对腐胺、腐生葡萄球菌对组胺和枯草芽孢杆菌对酪胺的抑制作用最为显著,减少程度分别为45.49%、57.54%、40.75%、51.03%、49.28%、64.33%。本研究为熏马肠以及其他发酵肉制品的提高安全性奠定了基础。     

Mixed starter cultures to control biogenic amine production in dry fermented sausages

Several combinations of an amine-negative Lactobacillus sakei strain, along with proteolytic Staphylococcus carnosus or Staphylococcus xylosus strains, were used to study the influence of mixed starter cultures on biogenic amine production during the manufacture of dry fermented sausages. Changes in pH, water content, proteolysis, microbial counts, and biogenic amine contents were simultaneously examined in a spontaneously fermented batch and in three mixed starter-mediated batches. A double-controlled microbial charge initially inoculated as mixed starter culture of L. sakei and Staphylococcus spp. (all amine-negative strains) drastically reduced tyramine, cadaverine, and putrescine accumulation. No production of other aromatic amines such as histamine, phenylethylamine, or tryptamine was observed in any batch. The polyamines, spermine and spermidine, were found in raw materials and their levels decreased slightly in the spontaneously fermented batch. No correlation between proteolysis and biogenic amine production was observed. The use of proper technological conditions favoring starter development and the use of the raw materials with good hygienic quality make it possible to produce fermented sausages nearly free of biogenic amines.     

Monitoring of biogenic amines in cheeses manufactured at small-scale farms and in fermented dairy products in the Czech Republic

The aim of the study was the monitoring of six biogenic amines (histamine, tyramine, phenylethylamine, tryptamine, putrescine, and cadaverine) and two polyamines (spermidine and spermine) in 112 samples of dairy products purchased in the Czech Republic, namely in 55 cheeses made in small-scale farms and in 57 fermented dairy products. The products were tested at the end of their shelf-life period. Neither tryptamine nor phenylethylamine was detected in the monitored samples; histamine was found only in four cheese samples containing up to 25 mg/kg. The contents of spermine and spermidine were low and did not exceed the values of 35 mg/kg. Significant amounts of tyramine, putrescine, and cadaverine occurred especially in cheeses produced from ewe’s milk or in long-term ripened cheeses. In about 10% of the tested cheeses, the total concentration of all the monitored biogenic amines and polyamines exceeded the level of 200 mg/kg, which can be considered toxicologically significant. In fermented dairy products, the tested biogenic amines occurred in relatively low amounts (generally up to 30 mg/kg) that are regarded safe for the consumer’s health.     

Biogenic amine accumulation in ripened sausages affected by the addition of sodium sulphite

The effect of two concentrations of sodium sulphite on biogenic amine accumulation during the ripening of slightly fermented sausages was examined. Three batches of sausages were manufactured: one without sulphite (control), one with 500 mg/kg and one with 1000 mg/kg of sodium sulphite. Considerable tyramine production (75–140 mg/kg dry matter [dm]) was observed in all batches. Sausages with sulphite, especially those with 500 mg/kg, accumulated higher amounts of tyramine than those without sulphite. Cadaverine was observed in the batch without sulphite (38 mg/kg dm), but its production was markedly inhibited by the presence of sulphite. A small amount of putrescine (4–12 mg/kg dm) was found during ripening and its final content was slightly higher in sausages with 1000 mg/kg of sulphite. No production of histamine, phenylethylamine or tryptamine was observed. Agmatine and spermine levels decreased during ripening, whereas spermidine levels remained constant. Although sausages with sulphite showed lower microbial counts, only cadaverine production was lower than in sausages without sulphite. In contrast, tyramine and putrescine production seemed to be stimulated by the presence of sodium sulphite.     

FORMATIONS OF BIOGENIC AMINES IN TURKISH STYLE SAUSAGE DURING RIPENING AND STORAGE PERIODS

Changes of biogenic amine (histamine, β‐phenylethylamine, serotonin, spermine, spermidine, cadaverine, putresine, 1,7‐diamino heptane, tryptamine and tyramine) concentrations and pH values of Turkish style sausages were investigated during ripening and storage. Cadaverine and β‐phenylethylamine were not detected during the ripening and storage periods for all sausage types. Storage time had a significant (P<0.05) effect on the formation of all biogenic amines. Starter culture usage decreased (P<0.05) biogenic amine formation. Histamine levels were higher in sausages prepared without additives and lower in the sausages made with starter culture than the sausages made without starter culture. There was no effect of starter culture and additives on formation of tryptamine, 1,7‐diamino heptane, serotonin or spermidine. Starter culture and additives affected (P<0.05) the formation histamine, putrescine and spermine. pH values of sausages sharply decreased (P< 0.05) from 5.98 to about 4.53 during 3 days of ripening. After that, pH values increased slowly and were nearly constant during the storage period.     

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.     

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.