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.     

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 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.     

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。     

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。     

Effect of low-dose irradiation on biogenic amines formation in vacuum-packed trout flesh (Oncorhynchus mykiss)

The effects of vacuum packaging followed by high-energy electron beam irradiation on the shelf-life of fillets of rainbow trout (Oncorhynchus mykiss) were examined. Samples were irradiated at doses of 0.25, 0.50, 0.75, 1.0 and 2.0 kGy. The control and treated packs were stored at 3.5 °C for up to 28, 42, 70 and 98 days (control - 0, 0.25, 0.50 and >0.50 kGy respectively). The content of seven biogenic amines (putrescine, cadaverine, spermidine, spermine, histamine, tyramine and tryptamine) were determined. Putrescine, cadaverine and tyramine showed very good correspondence with the irradiation dose and organoleptic properties. Samples of good quality contained less than 10 mg/kg of each of these amines. The polyamines spermidine and spermine did not show statistically significant changes with the irradiation dose and the time of storage. With few exceptions, histamine was not detected in the samples treated with radiation. Tryptamine was not detected in any of the samples.     

A survey of biogenic amines in chinese red wines

Eight biogenic amines were analyzed in 38 samples of red wine from five wine-making regions of China. The analysis was carried out by reverse phase high performance liquid chromatography with fluorescence detector and dansyl chloride precolumn derivation. Tryptamine was not found in any of the samples. Putrescine was detected in all samples (100%), followed by phenylethylamine (84.2%), spermidine (60.5%), histamine (57.8%), tyramine (57.8%), cadaverine (47.4%), and spermine (36.8%). In all the samples, the levels of aromatic and heterocyclic amines with toxicological effects were: 0–4.58 mg/l for phenylethylamine, 0–10.51 mg/l for histamine, and 0–9.13 mg/l for tyramine. The amount of histamine and tyramine in most of the samples (94.7%) was less than 8 mg/l. The amines associated with sanitary conditions were also found to be present in a very low range, between 0 and 12.98 mg/l for putrescine, 0 and 19.01 mg/l for cadaverine, respectively. In the case of other amines such as spermine and spermidine, they yielded very low levels varying between 0 and 2.64 mg/l for spermine, 0 and 3.82 mg/l for spermidine, respectively.     

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).     

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.     

Changes of yolk biogenic amine concentrations during storage of shell hen eggs

The RP-HPLC/UV method, using dabsyl derivatization, optimised for the determination of biogenic amines in egg yolk, was appropriate for quantification of putrescine, cadaverine, histamine, ethylamine, propylamine, ethanolamine, tyramine, tryptamine, spermine, spermidine, phenylethylamine. Detection limits ranged between 0.05 and 0.06 mg of biogenic amine/kg of egg yolk. Two experiments using, respectively, farm and avian eggs were conducted to evaluate yolk biogenic amine concentrations of fresh and stored eggs, and to explain the effect of temperature and time of storage in the levels of biogenic amines during egg shelf-life. Only five of the 11 biogenic amines under study were detected: putrescine, cadaverine, propylamine, ethylamine and ethanolamine. Storage time during shelf-life presented a significant effect on the levels of the five amines (p < 0.01). On the contrary, storage temperature did not presented a significant effect on the levels of the mentioned amines, p > 0.01. The significant reduction of biogenic amine concentration during the shelf-life justified the application of a multiple linear regression using stepwise method to estimate the storage time. The regression equation was applied with success to confirm the storage time of farm eggs and avian eggs that were stored at two different temperatures.     

Optimisation of a chromatographic procedure for determining biogenic amine concentrations in meat and meat products employing a cation-exchange column with a post-column system

The aim of this study was to optimise and validate a chromatographic method for determining biogenic amine (BA) in meat and meat products separated by a cation-exchange column with a post-column system, using o-phthalaldehyde as a derivatising reagent. A perfect separation of nine BA (tyramine, histamine, β-phenylethylamine, putrescine, cadaverine, tryptamine, agmatine, spermidine and spermine) was obtained in 22 min. The conditions were: column Tª 40 °C and coil Tª 45 °C, pump flow rate 0.8 mL/min, pH phase A 6.33, B 5.63 and C 13.00. The method was adjusted linearly in a range of 0.10–12 mg/L with a correlation coefficient superior to 0.998. Detection and quantification limits were between 0.03–0.10 mg/L and 0.10–0.20 mg/L, respectively. Precision studies were satisfactory, with RSD less than 2% and meat extracts recovering over 98%. This method showed an appropriate, precise, fast and versatile procedure for determining nine BA simultaneously in different meat product matrices.     

减盐对东北农家酱中生物胺形成的影响

目的：研究减盐对东北农家酱中生物胺形成的影响。方法：对正常食盐添加量与减盐条件（在正常食盐添加量的基础上分别减少10%、20%、30%、40%）下的东北农家酱中总胺与8 种生物胺（精胺、腐胺、色胺、苯乙胺、尸胺、组胺、酪胺、亚精胺）的含量进行分析,相应地对理化指标（氨基酸态氮、总酸含量）、微生物指标（菌落总数）进行分析,并进行感官品质评分。结果：与正常食盐添加量的对照实验组相比,随着食盐添加量的减少,减盐实验组中除精胺与亚精胺外,总胺与其他生物胺含量均有不同程度的增加,其中尸胺与组胺含量的增加最为明显。同时,减盐实验组中总酸含量、菌落总数及感官品质评分均有不同程度的变化;而氨基酸态氮含量则无明显变化。当减盐实验组中食盐添加量分别减少10%、20%时,总胺及各生物胺的含量无明显增加,且组胺含量分别为58.00、64.32 mg/kg,低于欧盟委员会的建议限量（100 mg/kg）;同时总酸含量、菌落总数无明显增加,但感官品质评分却有所升高。而当减盐实验组中食盐添加量分别减少30%、40%时,总胺及各生物胺含量明显增加,组胺含量分别为103.46、107.42 mg/kg,高于欧盟委员会的建议限量（100 mg/kg）;同时总酸含量、菌落总数明显增加,且感官品质评分则有所降低。结论：减盐东北农家酱生产时其食盐添加量在正常食盐添加量的基础上减少20%左右时比较适宜。     

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.     

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.     

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。     

A survey of biogenic amines in Chinese rice wines

Fourteen Chinese rice wines from four rice wine-making regions of China were analyzed for the first time, using HPLC with diode array detection after pre-column derivatization with dansyl chloride. The results showed that Chinese rice wines contained the five biogenic amines histamine, tyramine, cadaverine, spermine and spermidine. Histamine was detected in all samples (100%), followed by spermine (93%), cadaverine (87%), tyramine (79%) and spermidine (79%). The mean total level of biogenic amines in samples was 107 mg/l with a range from 39.30 to 241 mg/l. These levels are below the level that may elicit direct adverse reactions for most consumers. However, patients being treated with monoamine oxidase inhibitors should be aware of the presence of amines in rice wines and limit their consumption.     

Biogenic amines in Brazilian cheeses

The levels of biogenic amines in Brazilian cheeses were investigated for the first time. Ninety-two samples of 10 types of cheese were purchased at retail stores. The amines were extracted using hydrochloric acid, partitioned into diethyl ether, separated through reversed-phase ion-pair chromatography and quantified fluorimetrically after postcolumn derivatization with o-phthaldehyde. Overall, spermine was the most prevalent amine (93%), followed by histamine (65%), 2-phenylethylamine (62%), spermidine (61%), putrescine (60%), cadaverine (59%), serotonin (44%), agmatine (38%), tyramine (37%) and tryptamine (29%). Spermine, agmatine, 2-phenylethylamine, serotonin, spermidine and tryptamine were detected at low levels (< 4.10 mg/100g). Cadaverine, tyramine, histamine and putrescine were present at levels up to 111.00, 21.25, 19.65 and 17.37 mg/100g, respectively. There was variability on the type and levels of amine in each kind of cheese suggesting that amine formation and accumulation in cheese could be prevented. Efforts should be made to understand amine formation in cheese in order to optimize technology and secure low amine levels. Susceptible individuals should be advised to consume cheeses with low biogenic amines contents.     

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.     

新疆赤霞珠葡萄酒发酵过程中生物胺含量的变化

利用高效液相色谱技术分析葡萄酒发酵过程中8种生物胺含量的变化。结果表明,葡萄酒发酵过程中8种生物胺总含量整体呈上升趋势。色胺含量最高,最高可达到98.03 mg/L;腐胺和组胺含量较低,含量均不超过2.48 mg/L;精胺和亚精胺含量变化相对平稳,波动范围在1.81～3.07 mg/L之间,除色胺外,其他生物胺含量均不超过4.50 mg/L。苯乙胺、尸胺、酪胺等在酒精发酵后期和苹果酸-乳酸发酵初期大幅上升,在苹-乳发酵后期,酪胺和亚精胺含量有所下降,其他生物胺含量均有一定程度上升。     

Content and distribution of biogenic amines in Dutch-type hard cheese

Content of biogenic amines (BA; histamine, tyramine, tryptamine, putrescine, 2-phenylethylamine, cadaverine, spermidine, spermine) and counts of microorganisms (total aerobic and facultative anaerobic mesophilic microorganisms, TAC; lactic acid bacteria, LAB; coliforms; total anaerobic microorganisms, TAN; yeasts + moulds), were evaluated within the ripening interval of 1–6 months in Dutch-type hard cheese, produced by two different producers (the cheeses designated as R30 and R45) using pasteurized milk and different starter cultures (designated as R30-FD and R30-CH in the case of the R30 cheeses). The cheeses from the two producers differed in their fat content (30% and 45% of fat in dry matter, respectively).