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).     

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.     

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.     

Determination of biogenic amine profiles in conventional and organic cocoa-based products

Cocoa contains many compounds such as biogenic amines (BAs), known to influence consumer health. Spermidine, spermidine, putrescine, histamine, tyramine, β-phenylethylamine, cadaverine and serotonine have been found in several cocoa-based products using HPLC with UV detection after derivatisation with dansyl-chloride. Once optimised in terms of linearity, percentage recovery, LOD, LOQ and repeatability, this method was applied to real samples. Total concentrations of BAs ranged from 5.7 to 79.0 µg g^?1 with wide variations depending on the type of sample. BAs present in all samples were in decreasing order: histamine (1.9–38.1 µg g^?1) and tyramine (1.7–31.7 µg g^?1), while putrescine (0.9–32.7 µg g^?1), spermidine (1.0–9.7 µg g^?1) and spermidine (0.6–9.3 µg g^?1) were present in most of the samples. Cadaverine, serotonine and β-phenylethylamine were present in a few samples at much lower concentrations. Organic samples always contained much lower levels of BAs than their conventional counterparts and, generally speaking, the highest amounts of BAs were found in the most processed products.     

Formation of biogenic amines during the maturity process of raw meat products, for example of cervelat sausage

The raw meat product cervelat sausage with a diameter of 90 mm was investigated during maturation and storage over a period of about 12 weeks. The examination was carried out by ion exchange chromatography (amino acid analyzer) and included the amines ethylamine, propylamine, butylamine, putrescine, histamine, cadaverine, tyramine, β-phenylethylamine, spermidine and spermine. In addition, the starter bacteria cultures varied according to manufacturer. Over the duration of investigation, five different production batches were observed in parallel. During the maturity process and storage, after 87 days, tyramine (58.9 mg kg⁻¹) showed a clear dominance, followed by spermine (34.1 mg kg⁻¹), putrescine (26.1 mg kg⁻¹) and spermidine (8.7 mg kg⁻¹), if starter bacteria culture 1 was used. Other amines were not detected. The use of starter bacteria culture 2 led, after 87 days, to the highest content of putrescine (244.1 mg kg⁻¹), followed by tyramine (119.2 mg kg⁻¹), histamine (52.9 mg kg⁻¹), spermine (37.5 mg kg⁻¹) and spermidine (9.2 mg kg⁻¹). With the use of starter bacteria culture 3, for spermine and spermidine a comparable behavior was shown to that with culture 1 or 2. After 87 days, putrescine reached the highest value of 484.2 mg kg⁻¹, followed by tyramine (119.4 mg kg⁻¹) and histamine (111.0 mg kg⁻¹). Cadaverine (36.9 mg kg⁻¹) could also be determined. As an accompanying investigation, the parameters pH, water activity (a_w) and microbial count (lactobacilli, enterobacteria, staphylococci and yeast) were determined. An investigation of the sections center-middle-edge showed that the content of amines increases to its highest values in the middle. The edge showed the lowest content. This behavior correlates with the a_w distribution of a raw sausage.     

Salting-out assisted liquid–liquid extraction with dansyl chloride for the determination of biogenic amines in food

Biogenic amines (BAs) are compounds present in foodstuff of animal origin that not only can be harmful to the consumer but can also serve as indicators of the product's edibility status. However, BAs’ analysis is rather complicated. Derivatisation methods have been employed to make their analysis less challenging. Salting-out assisted liquid–liquid extraction (SALLE) was herein applied to decrease time that the derivatising agent dansyl chloride (DNS-Cl) requires to be effective. DNS-Cl derivates were separated and analysed by high-performance liquid chromatography with UV and fluorescence detection (HPLC-UV/FD). It was by this way possible to analyse methylamine, dimethylamine, ethylamine, β-phenylethylamine, isopentylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine in a straightforward manner in several fish-based and meat-based samples, obtaining results in the order of magnitude of about a few mg per kg. All calibrations curves had r² higher than 0.99, and limits of detection ranged from 0.0075 to 1.6 mg L⁻¹.     

Analysis of Vacuum Packed Beef Regarding Psychrotrophic Bacteria Growth and Biogenic Amines Content

It has been recognized that biogenic amines (BA) content in meat can be considered a freshness marker. Considerable amounts of some BAs can appear during food storage under certain conditions, according to the handling of the raw material, technology applied, storage temperature and time, packaging condition, mainly if amino acid - decarboxylase positive microorganisms are present. The aim of this study was to evaluate the psychrotrophic bacteria growth and metabolic production of BAs during chill storage of beef. The vacuum packed beef cuts (Longissimus dorsi muscle) were analyzed during storage at 7 °C at 0, 15, 30, 45, and 60 d, to determine the psychrotrophic bacteria growth and the BAs amount. The BAs considered were: putrescine, cadaverine, histamine, spermidine, and spermine. The BAs quantitative determination was carried out by reversed phase high-performance liquid chromatography (HPLC) with UV detection. Statistic procedures were performed using SAS statistical software. The growth parameters of psychrotrophic bacteria including lag phase, maximum specific growth rate, maximum bacterial cell density, initial population, mean square error, and coefficient of determination were determined according to Baranyi and Roberts model. The values of histamine and spermidine increased significantly (P < 0.0001) during storage time, while the levels of spermine decreased (P < 0.0001). Psychrotrophic bacteria counts increased significantly (P < 0.0001) reaching 7.6 log cfu/g over time. The counts of this group positively correlated to histamine and spermidine (r = 0.68 and 0.61, respectively), while spermine showed a negative correlation (r = -0.70). Conversely, no significant correlation was found between psychrotrophics counts and putrescine or psychrotrophics counts and cadaverine.     

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。     

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。     

冷藏期内猪肉肌苷酸与生物胺两者变化关系的研究

采用高效液相色谱法测定生鲜猪肉的背最长肌在4℃温度条件下肌苷酸(IMP)和腐胺、尸胺、组胺、酪胺、亚精胺、精胺这六种生物胺含量的变化情况,从而进一步分析它们之间的相关性。结果表明,随着货架期的延长,IMP含量呈先上升后降低的趋势,并在第2d达到最高;精胺含量基本保持在4.0mg/kg左右,组胺的含量始终很低,其它的胺类物质都有所增加,尸胺的变化最为突出;虽然腐胺和亚精胺的含量比较低,但是仍然有明显的变化;酪胺变化也非常明显。从Person积差相关系数可以看出,IMP与其他指标相关系数呈负相关显著,有的指标是不显著的;在0.01水平上,IMP与亚精胺之间的负相关性最强,达到了-0.981;其次是尸胺与IMP之间,相关系数是-0.960,呈显著负相关;酪胺也与IMP显著负相关;IMP与腐胺、组胺、精胺的相关系数都不显著。因此,通过测定IMP含量变化可以预测猪肉新鲜度,且可作为猪肉保藏与加工过程中品质控制的重要指标之一。     

Determination of biogenic amines in Japanese miso products

Eight miso products sold in supermarket were purchased from Japan and analyzed for the determination of biogenic amines (BAs). Quantitative determination of BAs in miso products was carried out by means of HPLC with spectrophotometric UV-VIS detector. The amines were extracted in acid aqueous solution (HClO₄) and then derivatized by dansyl chloride. The levels of pH in the tested miso samples were ranged from 4.8–5.5. The BAs (tryptamine, 2-phenyl ethylamine, putrescine, cadaverine, agmatine, tyramine, and spermidine) determined in 8 miso samples were found in their respective range of ND-0.13, ND-4.20, ND-1.29, ND-0.87, ND-0.90, ND-4.19, and ND-3.57 mg%. None of the miso samples contained histamine and spermine, the major BAs of food poisoning.     

The effects of nanoemulsions based on citrus essential oils on the formation of biogenic amines in trout fillets stored at 4 ± 2°C

The influences of nanoemulsions based on citrus peel (orange, lemon, mandarin, and grapefruit) essential oils on the biogenic amine (BA) formation in rainbow trout (Oncorhynchus mykiss) fillets stored 4 ± 2°C were investigated. The results demonstrated that main BAs in the trout fillets were putrescine, cadaverine, spermidine, spermine, serotonin, tyramine, dopamine, and agmatine. It was determined that nanoemulsion application had generally suppressive effect on the formation of BAs during storage. It was also found that especially orange and lemon nanoemulsion groups were effective on histamine formation. Histamine values ranged from 0.00 to 4.08 in all groups during storage and the levels were below the limit values of FDA and EU throughout the storage period for all treatment groups. Similarly, Quality Index (QI) values of BAs were found to be below legal limits in all groups. Consequently, it was found that nanoemulsions prepared from essential oils kept the BA formation at a lower level during storage.     

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.     

Relation of biogenic amines to microbial and sensory changes of precooked chicken meat stored aerobically and under modified atmosphere packaging at 4 °C

The formation of biogenic amines and their correlation to microflora and sensory characteristics of a precooked chicken meat product stored aerobically and under modified atmosphere packaging (MAP) (30% CO₂, 70% N₂) was studied. Putrescine was the main amine formed both in aerobically and MA-packaged chicken samples. For the rest of the biogenic amines, including tyramine, histamine, and cadaverine, a stepwise increase was recorded throughout the 23-day storage period under the above packaging conditions. Spermidine was found in higher amounts, as compared to spermine in both aerobically and MA-packaged chicken samples at 4 °C. Formation of these amines in precooked chicken stored either aerobically or under a 30% CO₂, 70% N₂ atmosphere followed an inconsistent trend during the entire storage period at 4 °C. Agmatine, β-phenyl-ethylamine, and tryptamine were not detected in precooked chicken. Of the bacterial groups monitored, lactic acid bacteria (LAB) became the dominant bacteria after day 8 of storage under MAP while LAB were the dominant population of natural microflora of precooked chicken stored both aerobically or under MAP, reaching 7.5 and 8.0 log cfu/g, respectively, on day 23 of refrigerated storage. Enterobacteriaceae populations in chicken meat were below the detection limit (<1 log cfu/g) by pour plating throughout the 23-day storage period, irrespective of packaging conditions. Based on sensory data, after ca. 8 days for the precooked chicken meat stored aerobically and after 12 days under MAP (time to reach initial decomposition stage, score of 2) the putrescine and tyramine content of chicken samples were ca. 14–19 and 1.4 mg/kg, values that may be proposed as the limit for spoilage initiation of precooked chicken meat (respective TVC for both aerobically and MA-packaged chicken meat were ca. 6.5 log cfu/g).     

Production of biogenic amines by lactic acid bacteria and bifidobacteria isolated from dairy products and beer

The aim was to monitor production of eight biogenic amines (BAs) (histamine, tyramine (TYR), tryptamine, putrescine, cadaverine (CAD), phenylethylamine, spermine and spermidine) by selected 81 lactic acid bacteria (LAB) strains: Lactobacillus, Lactococcus, Leuconostoc, Enterococcus, Pediococcus, Tetragenococcus and Bifidobacterium. The tested LAB and bifidobacteria were isolated from dairy products and beer. The decarboxylase activity of the micro‐organisms was studied in growth medium after cultivation. The activity was monitored by HPLC after the pre‐column derivatisation with dansylchloride. Fifty LAB showed decarboxylase activity. Thirty‐one strains produced low concentrations of CAD (≤10 mg L^?1). Almost 70% of beer isolates generated higher amounts of TYR (≤3000 mg L^?1). Most of the tested LAB demonstrated decarboxylase activity. The above micro‐organisms can contribute to the increase of content of BAs in dairy products or beer and thereby threaten food safety and health of consumers. Production of BAs even by the representatives of some probiotic strains (Bifidobacterium and Lactobacillus rhamnosus) was detected in this research. This study has also proved that contaminating LAB can act as sources of higher amounts of CAD and TYR in beer.     

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.     

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).     

A comparative study for determination of biogenic amines in meat samples by capillary isotachophoresis with two electrolyte systems

The selected biogenic amines, BAs (putrescine, cadaverine, histamine, tyramine, spermidine and spermine), in pork, beef, and poultry meat samples were determined by two simple procedures without derivatization using isotachophoretic technique. Two electrolyte systems were proposed for their separation and determination with satisfactory separation parameters. The linearity of the proposed procedures was between 2–10 mg L^?1 and 5–50 or 10–60 mg L^?1, whereas average recoveries for the standard solutions of amines and their mixture varied between 99–100 % and 95–105 %, respectively. Average recoveries for standard addition method (96–99 %) indicated satisfactory accuracy of proposed electrolyte systems. The obtained coefficient of variation values for standard solutions and meat samples ranged from 0.62 to 4.54 % and from 0.56 to 6.03 %, respectively, which confirmed good repeatability. Except for spermidine and spermine, BAs levels in fresh meat samples were generally low. The effect of storage time on the content of biogenic amines in meat was also studied indicating the increase of BAs levels (excluding Spermine and Spermidine).     

Formation of biogenic amines by Gram-negative bacteria isolated from poultry skin

The aim of this study is to explore production of biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) by 88 Gram-negative bacteria isolated from poultry skin (41 isolates of family Enterobacteriaceae, 21 isolates of genus Aeromonas, 16 isolates of genus Pseudomonas, and 10 isolates of other Gram-negative rods). Ion-exchange chromatography was employed to analyse the above mentioned amines. Enterobacteria were found to be the largest producers of amines with proved presence of tyramine, agmatine, putrescine, and cadaverine in cultivation broth after incubation of bacteria. Putrescine and cadaverine were the most abundant products. Presence of at least two biogenic amines, i.e. mainly concurrent presence of putrescine and cadaverine, was revealed in 19 enterobacteria strains. Eleven isolates classified into Aeromonas genus produced putrescine and five of them also formed cadaverine. The other observed biogenic amines (histamine, spermine and spermidine) were not found among tested isolates. Production of biogenic amines by any Pseudomonas family isolates was not proved.     

Studies of levels of biogenic amines in meat samples in relation to the content of additives

The impact of meat additives on the concentration of biogenic amines and the quality of meat was studied. Fresh white and red meat samples were fortified with the following food additives: citric and lactic acids, disodium diphosphate, sodium nitrite, sodium metabisulphite, potassium sorbate, sodium chloride, ascorbic acid, α-tocopherol, propyl 3,4,5-trihydroxybenzoate (propyl gallate) and butylated hydroxyanisole. The content of spermine, spermidine, putrescine, cadaverine, histamine, tyramine, tryptamine and 2-phenylethylamine was determined by capillary isotachophoretic methods in meat samples (fresh and fortified) during four days of storage at 4°C. The results were applied to estimate the impact of the tested additives on the formation of biogenic amines in white and red meat. For all tested meats, sodium nitrite, sodium chloride and disodium diphosphate showed the best inhibition. However, cadaverine and putrescine were characterised by the biggest changes in concentration during the storage time of all the additives. Based on the presented data for the content of biogenic amines in meat samples analysed as a function of storage time and additives, we suggest that cadaverine and putrescine have a significant impact on meat quality.