发酵香肠中生物胺的研究

本文采用RP-HPLC法测定了发酵香肠中的生物胺(苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺、精胺)的含量。样品经0.4mol/L高氯酸溶液提取,丹磺酰氯衍生,流动相为乙腈和水,采用梯度洗脱,流速为1mL/min,紫外检测波长为254nm。该方法检测限为:腐胺、尸胺、亚精胺、酪胺和精胺为0.1μg/mL,组胺0.5μg/mL,苯乙胺0.05μg/mL。回收率分别为苯乙胺86.71%、腐胺88.88%、尸胺94.55%、组胺87.57%、酪胺83.67%、亚精胺88.55%、精胺94.91%。结果表明发酵香肠中生物胺的种类及含量因香肠的品种而异,7种生物胺平均总量为13.40mg/100g,变异范围为7.83~19.13mg/100g。本法简便、快速、灵敏、可靠。     

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。     

六类鱼制品中生物胺的HPLC法测定

针对不同类型的鱼制品建立同时检测腐胺、尸胺、精胺、亚精胺、酪胺、苯乙胺、组胺和色胺8种生物胺含量的高效液相色谱(HPLC)法。应用该方法,测定了干燥制品、油炸制品、烘烤制品、发酵制品、混揉制品以及萃取制品6类鱼干制品中的生物胺含量。结果显示,萃取制品-鱼油中未检测出生物胺,其余5类产品中生物胺含量最高的为发酵制品(73.42~70.59 mg/kg)和混揉制品(55.48~49.88 mg/kg),其次为油炸制品(17.72~14.32 mg/kg),而鱼干制品(5.28~5.17 mg/kg)和烘烤制品(4.75~4.69 mg/kg)中的生物胺含量相对较少;另外,尸胺、组胺、精胺和亚精胺四种生物胺为除萃取制品之外的5类鱼制品中普遍存在的4种生物胺成分;其中尸胺是鱼制品中变化最大、影响最为明显的单体生物胺,检测产品中尸胺含量有利于监控、评判鱼干制品的质量。     

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。     

酿酒工艺对葡萄酒中生物胺的影响

探讨了不同酿酒工艺对葡萄酒中生物胺(组胺、苯乙胺、酪胺、色胺、腐胺、尸胺、精胺和亚精胺)含量的影响。结果显示,葡萄酒酿造过程中生物胺主要在酒精发酵和苹果酸-乳酸发酵过程产生。酒精发酵过程生成量较少,主要产生腐胺和精胺,果胶酶的使用、酵母接种量、发酵温度等因素可以调节酒精发酵过程生物胺的生成量;乳酸菌是葡萄酒生物胺最主要来源,在苹果酸-乳酸发酵过程中会产生大量的组胺和色胺。     

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。     

高盐稀态酱油生产过程中的生物胺变化规律

采用丹磺酰氯柱前衍生高效液相色谱法测定4种市售高盐稀态酱油中色胺、苯乙胺、腐胺、尸胺、组胺、酪胺、亚精胺、精胺8种生物胺的含量,并监测了其中一种酱油生产过程中生物胺含量和生物胺产生相关指标的动态变化。结果显示,市售酱油总生物胺含量差别较大,范围为102.87~760.28 mg/kg,但均以酪胺、组胺和苯乙胺为主。酱油发酵过程中,大部分生物胺表现出相似的变化趋势,即发酵前期快速升高,在50~60 d左右达到峰值,再逐渐降低,酪胺和组胺是酱油发酵过程中的主要生物胺,最高含量分别达到2 417.36 mg/kg和815.89mg/kg。氨基酸态氮前期变化趋势与总生物胺含量变化基本一致,之后趋于稳定;p H值降低有利于生物胺的形成,最低时生物胺含量达到峰值。结果表明,在市售酱油和不同发酵时期酱油中生物胺的含量情况不容乐观,需加以研究与关注。     

不同发酵剂对发酵香肠中风味物质释放及有害生物胺控制的影响

通过接种植物乳杆菌与肉葡萄球菌,将试验分为复合发酵组和单一植物乳杆菌组,并以自然发酵为对照。探究发酵剂对发酵香肠加工过程中pH、Aw、色泽、亚硝酸盐、风味物质释放及有害生物胺含量控制的影响。结果表明:复配组产酸速率单一组产酸速率对照组产酸速率,且干燥结束时复配组降到组中最低为4.74;pH快速降低也使复配组Aw下降速率显著快于其它两组(P0.05);9 d后成熟结束时,复配组亚硝酸盐含量为12.6 mg/kg,显著低于其它两组及国家安全规定30 mg/kg(P0.05);从3组香肠中检出尸胺、β-苯乙胺、酪胺、组胺及精胺和亚精胺,由于精胺和亚精胺是肉中固有生物胺且含量变化微小,所以在试验中未加以说明;成品时所列4种生物胺总量在33.37～40.41 mg/kg范围,复配组中组胺及酪胺含量分别为2.89,22.54 mg/kg,低于美国食品及药品管理局(FDA)规定标准(组胺含量50 mg/kg,酪胺含量≤100 mg/kg)(P0.01)。从香肠中共检出26种挥发性风味物质,其中添加微生物发酵组风味物质总含量显著高于对照组(P0.01)。综上,复配发酵剂可使pH、Aw快速降低,有效抑制和降低香肠残留亚硝酸盐及有害生物胺含量,对香肠中风味物质释放和含量增加起主要作用。     

宣威火腿加工过程中生物胺变化规律

用40条重量在9～11 kg的原料腿按照传统工艺加工宣威火腿,分别在腌制前(24 h)、腌制结束(18d)、风干结束(125 d)、成熟中期(250 d)与成熟结束(360 d)这5个加工阶段随机取出5条火腿,以股二头肌为中心取约100 g样品,以HPLC方法测定生物胺含量。结果表明,宣威火腿样品中共检测出7种生物胺,分别是色胺、苯乙胺、腐胺、酪胺、尸胺、亚精胺和精胺。在各个加工阶段,精胺的含量都是最高的,其次是酪胺与亚精胺。酪胺、亚精胺与精胺的含量在5个加工阶段都没有显著变化(P>0.05)。苯乙胺与腐胺含量显著增加(P<0.05),在成熟中期(250 d)时达最高值,随后含量有所降低(P>0.05)。生物胺总量随着加工过程的进行有所增加,在成熟中期(250 d)达最大值11.51mg/100 g,随后生物胺总量有降低的趋势,但差异不显著(P>0.05)。     

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.     

Levels of biogenic amines in retail market fermented meat products

The qualitative and quantitative profile of biogenic amines (BA) in 50 samples of dry fermented sausages sold in Greek retail markets were determined by HPLC. Putrescine, cadaverine, tryptamine, β-phenylethylamine spermidine, spermine were analysed by UV detection after pre-column derivatization with benzoyl chloride, whereas tyramine and histamine were analysed by fluorescence detection after post-column derivatization with o-phthaldialdehyde (OPA). With the exception of spermidine and spermine a wide variation of BA levels was observed among the samples. Of the BA examined, tyramine, putrescine, histamine and cadaverine showed high concentrations ranging from: 0 to 510 mg/kg (median: 197.7 mg/kg), 0 to 505 mg/kg (median: 96.5 mg/kg), 0 to 515 mg/kg (median: 7.0 mg/kg) and 0 to 690 mg/kg (median: 3.6 mg/kg), respectively. The histamine content of 28% of the samples exceeded the toxicity limit of 100 mg/kg set for histamine in some fish species. Levels of tryptamine and β-phenyl-ethylamine never exceeded 50 and 29 mg/kg, respectively. Results of the present study suggest that the amounts of BA in dry fermented sausages, sold in Greek retail markets, may pose a potential health risk for sensitive individuals or for those undergoing classical monoamine oxidase inhibiting (MAOI) drug therapy.     

Formation of biogenic amines and relation to microbial flora and sensory changes in smoked turkey breast fillets stored under various packaging conditions at 4 degrees C

The present study evaluated: (1) the formation of biogenic amines (BAs) in smoked turkey fillets during storage under aerobic and modified atmosphere packaging (MAP) conditions at 4 degrees C, (2) the relation of BAs to microbial and sensory changes in turkey meat and (3) the possible role of BAs as indicators of poultry meat spoilage. Smoked sliced turkey fillets were stored in air and under vacuum, skin and two modified atmospheres (MAP), M1 (30% CO(2)/70% N(2)) and M2 (50% CO(2)/50% N(2)), at 4+/-0.5 degrees C, for a period of 30 days. The BAs determined were: tryptamine, tyramine, histamine, putrescine, cadaverine, spermidine and spermine. Low levels of BAs were observed throughout the entire storage period, with the exception of histamine, tyramine and tryptamine, for which higher concentrations were recorded. Values for these three BAs were the highest for air-packaged samples (32.9, 25.0 and 4.1mg/kg, respectively) and the lowest for skin-packaged samples (11.9, 4.3 and 2.8 mg/kg, respectively) after 30 days of storage. All microorganism populations increased throughout the storage period, except for Pseudomonas spp. and Enterobacteriaceae, in skin-packaged fillets and modified atmosphere M2, which remained under the method detection limit (<1logCFU/g) until day 30 of storage. Pseudomonas spp. and Enterobacteriaceae for the rest of the packaging treatments remained below 5logCFU/g throughout storage. On the other hand, lactic acid bacteria were dominant throughout the storage period, regardless of the packaging conditions reaching 8.9logCFU/g on day 30 of storage. Mesophiles reached 7logCFU/g after ca. 19-20 days for the air and skin packed samples, 22-23 days for the M2 and vacuum packed samples and 25-26 days for the M1 packed samples. BA values for tryptamine, histamine and tyramine correlated well with both microbiological and sensory analyses data. Tryptamine, histamine and tyramine may be used as chemical indicators of turkey meat spoilage.     

Relation of biogenic amines with microbial and sensory changes of whole and filleted freshwater rainbow trout (Onchorynchus mykiss) stored on ice

The biogenic amine (BA) content of whole and filleted rainbow trout was monitored during ice storage for a period of 18 days and related to respective microbial and sensorial changes occurring during the same period. Eight amines, namely, putrescine, cadaverine, tyramine, spermidine, tryptamine, beta-phenylethylamine, spermine, and histamine, were determined. Agmatine was not detected in any of the fish samples. In all cases, concentration of BAs was higher (P < 0.05) in filleted compared with whole trout samples. Pseudomonads, H2S-producing bacteria, and, to a lesser extent, Enterobacteriaceae were the dominant microorganisms in both whole and filleted trout. Higher populations (P < 0.05) of these microorganisms were present in filleted trout compared with whole fish samples. Of the BAs determined, concentration of putrescine, cadaverine, spermidine, tryptamine, and beta-phenylethylamine increased steeply in both whole and filleted trout between days 15 and 18 of storage when pseudomonads and H2S-producing bacteria reached approximately 10(6) to 10(7) CFU/g. For the rest of the BAs, including tyramine, histamine, and spermine, a stepwise increase was recorded throughout the entire storage period. Interestingly, Enterobacteriaceae counts remained below 10(6) throughout the entire storage period, accounting for the lower production of histamine. A putrescine value of 13 to 14 mg/kg and a spermidine value of approximately 7 mg/kg for both the whole and filleted trout obtained after 12 and 9 days, respectively, may be proposed as the upper limit for spoilage initiation (freshness indicator) of fresh rainbow trout based on sensorial and microbiological (total viable count of 10(6) to 10(7)) data. With respect to other amines determined, both tyramine and spermine may also be proposed as freshness indicators preferably for whole trout, whereas tryptamine, beta-phenylethylamine, histamine, and cadaverine produced only during later stages of storage are not suitable as freshness indicators of either whole or filleted trout.     

我国传统干酪中生物胺含量检测分析

通过反相高效液相色谱法对来自内蒙古、新疆、西藏、云南、北京市场共17种中国传统干酪样品生物胺含量进行了分析。结果表明,反相高效液相色谱法能准确地测定干酪中生物胺含量;在全部干酪样品中,8种生物胺含量大小不一,生物胺总量在1.30～41.27 mg/kg范围内,均低于国家标准规定要求,其中有14个样品的生物胺总量低于10 mg/kg,第10号干酪生物胺含量为41.27 mg/kg,为所有样品中含量最高,其次为第1号样品和第9号样品,分别为18.48 mg/kg和10.35 mg/kg。相关性分析结果显示,色胺与苯乙胺,腐胺与尸胺、酪胺、亚精胺,组胺与精胺,酪胺与精胺之间在0.01水平上具有显著相关性。     

Residual nitrite and biogenic amines of traditional northeast sauerkraut in China

The safety of northeast sauerkraut (NS) has always been a concern in China. In the present study, the nitrite and biogenic amine (BA) contents in 378 NS, collected or purchased from Chinese families and markets (including morning markets and local supermarkets), were measured. The results showed that the nitrite and BA contents of samples from Chinese families and some morning markets were higher than those from local supermarkets. The highest level of nitrites was 38.5 ± 5.2 mg/kg in unpackaged NS from Chinese families, and the lowest was 17.2 ± 2.1 mg/kg in packaged NS from local supermarkets. The highest levels of tyramine and tryptamine were 203 ± 3 and 28.1 ± 4 mg/kg in unpackaged NS from Chinese families, respectively. The highest levels of histamine, putrescine, and cadaverine were mainly found in Chinese families and morning markets, and the lowest level of BAs was from local supermarkets. The results suggest that improving the safety of NS by controlling environmental sanitation and strengthening standardised production is a top priority at present in China.     

Detection of biogenic amine producer bacteria in a typical Italian goat cheese

The aim of this study was to research decarboxylating bacterial strains and biogenic amine content in a typical Italian goat cheese (Robiola di Roccaverano). The study was performed on fresh and ripened samples of goat cheese manufactured from industrial and artisanal producers. Sixty-seven bacterial strains isolated showed decarboxylating activity, and Enterococcus faecalis was the most widespread decarboxylating species in all artisanal and industrial products. Pediococcus acidilactici and Enterococcus malodoratus were also identified as biogenic amine producers in Robiola di Roccaverano cheese. All the E. faecalis strains isolated in this study were able to decarboxylate tyrosine. Tyramine was the most abundant biogenic amine in cheese samples, while histamine was the most widespread. High amounts of these two biogenic amines were found in ripened samples (up to 2,067 mg/kg for tyramine and 1,786 mg/kg for histamine), whereas 2-phenylethylamine and tryptamine were present in almost all ripened cheeses at low concentrations. The detection of strains producing biogenic amines and the high concentrations of tyramine and histamine found in ripened Robiola di Roccaverano could represent a potential risk to the consumer.     

Biogenic amines in selected foods of animal origin obtained from the Croatian retail market

ABSTRACT

This study aimed to investigate the presence of eight biogenic amines (BAs): tryptamine (TRP), phenylethylamine (PHE), putrescine (PUT), cadaverine (CAD), histamine (HIS), tyramine (TYR), spermidine (SPD) and spermine (SPM) in cheese, fish & fishery products and meat & meat products obtained from the Croatian retail market. A selective and robust method of high-performance liquid chromatography (HPLC) with diode array detection (DAD) was applied for the determination of BAs in a total of 91 samples in accordance with the performance criteria outlined in the European legislation. A high inter- and intra-food group variability of the amounts of BAs was observed. In the analysed samples, the most represented amines were TYR, HIS, CAD and PUT. Based on the highest content of the most toxic BAs (HIS and TYR) and consequential food safety concerns, the studied food groups can be ranked in the following order: cheese (HIS up to 106.4 mg/kg; TYR up to 206.6 mg/kg), fish &fishery products (HIS up to 98.8 mg/kg; TYR up to 47.9 mg/kg), and meat & meat products (HIS up to 20.0 mg/kg; TYR up to 117.5 mg/kg). The total BA content was significantly higher (p < .05) in fermented in comparison with other food. The study aimed to contribute to the knowledge on BA toxicity and food quality, as well as to support the indispensable future studies of consumption data and exposure assessment, to the end of defining allowable BA concentrations in food.     

Identification and Estimation of Histamine, Tryptamine, Phenethylamine and Tyramine in Soy Sauce by Thin-Layer Chromatography of Dansyl Derivatives

Histamine, tryptamine, phenethylamine and tyramine in soy sauce were derivatized by reaction with dansyl chloride in acetone-water (3:l). The dansyl derivatives were extracted with benzene, separated by TLC, and quantitated by measuring the fluorescence of each derivative. The average recovery for the entire procedure was 80% for histamine and over 87% for the other three amines. Tyramine was the chief amine found. The constancy of tyramine formation was demonstrated by its presence in measurable quantities in 12 of 15 soy sauce samples examined. The highest tyramine concentration (0.47 mg/ml) was found in a tamari soy sauce. The tyramine content averaged 0.18 mg/ml. Histamine, found in 10 of 15 samples, and tryptamine, found in only 5, averaged 0.1 mg/ml and 0.08 mg/ml respectively in the positive samples. Phenethylamine was not found in any of the 15 samples.     

Irradiation Effects on Biogenic Amines in Korean Fermented Soybean Paste During Fermentation

ABSTRACT: Irradiation effects on biogenic amines (BAs) and microbiological populations of Korean fermented soybean paste were investigated during fermentation. Soybean paste was prepared and irradiated with doses of 5,10, and 15 kGy, and then fermented at 25 °C for 12 wk. Bacillus spp. and lactic‐acid bacteria decreased by irradiation but increased during fermentation. Biogenic amines detected were putrescine, cadaverine, β‐phenylethylamine, spermidine, spermine, tryptamine, histamine, tyramine, and agmatine. A significant difference was not observed in BA content between control and irradiated samples immediately after gamma irradiation. However, 4 kinds of BAs, putrescine, tryptamine, spermidine, and histamine, showed significant reduction by irradiation during fermentation (P < 0.05).     

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。