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.     

Comparison of ELISA and HPLC methods for the determination of biogenic amines in commercial doenjang and gochujang

In order to conduct fast detection of biogenic amines (BAs) in doenjang and gochujang, a competitive direct enzyme-linked immunosorbent assay (CD-ELISA) for histamine was compared with HPLC analysis. The histamine contents were determined from 0.7 to 420 mg/kg by CD-ELISA and 2.2 to 382.4 mg/kg by HPLC. Both methods showed a good agreement for histamine analysis (R=0.969). In addition, the histamine contents by CD-ELISA were correlated to the sum of 7 major BAs in the samples (R=0.910), suggesting that the CD-ELISA can be used as a rapid indicator for total BAs in doenjang and gochujang.     

Determination of biogenic amines in <Emphasis Type="Italic">Cheonggukjang</Emphasis> using ultra high pressure liquid chromatography coupled with mass spectrometry

A rapid and reliable method for the simultaneous determination of 12 biogenic amines (BAs; agmatine, phenylethylamine, tyramine, putrescine, cadaverine, histamine, serotonin, tryptamine, spermidine, noreinephrine, dopamine, and spermine) in cheonggukjang was optimized and validated using ultra high pressure liquid chromatography-electrospray tandem mass spectrometry (UHPLC-ESI-MS/MS). The BAs were dansylated and separated on a C8 column under LC gradient of 7 min duration, and detected by MS/MS with the multiple reaction monitoring mode (MRM). This method exhibited excellent linearity for all of the analytes with correlation determination (R²) higher than 0.98. The limits of detection (LODs) were 10.8–39.6 μg/kg. The precision results were expressed as relative standard deviation (RSD), ranged from 0.3 to 14.3% for intra-day and from 0.9 to 15.4% for the interday. The proposed method will help to ensure for quality control by monitoring of BAs in cheonggukjang.     

Real time quantitative PCR detection of histamine-producing lactic acid bacteria in cheese: Relation with histamine content

Biogenic amines (BAs) are low molecular weight organic bases with biological activity, mainly formed by the microbial decarboxylation of amino acids. The consumption of food containing large amounts of some BAs can have toxicological consequences. Histamine is the most active BA and the most frequently involved in food-borne intoxications. This article reports the concentrations of histamine found in 80 cheeses made from different types of milk and subjected to different ripening periods. Histamine was detected in 41.25% of the samples, with values ranging from 20 to over 1000 mg per Kg of cheese. The highest histamine concentrations were recorded in long ripening cheeses made from raw milk. The presence and quantification of histamine-producing lactic acid bacteria was determined by RT-qPCR, and a good association was obtained between the C_t values and histamine concentrations determined by HPLC.     

Identification and characterisation of headspace volatiles of fish miso, a Japanese fish meat based fermented paste,with special emphasis on effect of fish species and meat washing

The present study investigated the suitability of four species of trash fishes for the production of fish miso, a Japanese fermented fish meat paste compared with soy and rice miso from the point of view of product aroma. The effect of washing fish meat on finished product was also evaluated. Headspace volatiles for different miso samples were analyzed by using solid phase microextraction (SPME) technique. A total of 107 volatile compounds have been identified, where 94 were common for all the miso samples. Considering the lower threshold perception and higher odour active values 2-methylbutanal, 3-methylbutanal, methional, isoamyl acetate, dimethyl disulfide, dimethyl trisulfide, 2,3-butanedione, 3-methylethyl butanoate, 3-methyl-1-butanol, ethyl hexanoate, 1-octen-3-ol, heptanol, heptanal and 2-undecanone were identified as key compounds for the miso products. Principal components analysis (PCA) and hierarchical cluster analysis (HCA) of headspace volatiles clearly elucidated the relationship amongst different miso samples based on fish species and effect of fresh water washing of meat on aroma of finished product.     

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).     

Development and validation of a solid-phase extraction method coupled with HPLC-UV detection for the determination of biogenic amines in Chinese rice wine

ABSTRACT

A reliable and accurate method for the determination of seven biogenic amines (BAs) was developed and validated with Chinese rice wine samples. The BAs were derivatised with dansyl chloride, cleaned up using solid-phase extraction (SPE) and separated by high-performance liquid chromatography (HPLC) coupled with ultraviolet (UV) detection. The optimised derivatisation reaction, conducted at pH 9.6 and 60°C for 30 min, ensured baseline separation and peak symmetry for each BA. SPE clean-up using Oasis MCX cartridges yielded good recovery rates for all BAs and effectively reduced matrix effects. The developed method shows good linearity with determination coefficients of more than 0.9989 over a concentration range of 0.1–100 mg l^?1. The limits of detection (LODs) for the investigated BAs ranged from 2.07 to 5.56 µg l^?1. The intra- and inter-day relative standard deviations (RSDs) ranged from 0.86% to 3.81% and from 2.13% to 3.82%, respectively. Spiking experiments showed that the overall recovery rates ranged from 85% to 113%. Thus, the proposed method was demonstrated as being suitable for simultaneous detection, with accurate and precise quantification, of BAs in Chinese rice wine.     

水产品中生物胺的检测方法研究进展

生物胺是一类低分子量含氮有机化合物,主要是由氨基酸进行脱羧反应而产生,广泛存在于各类食品中,尤其是蛋白质含量较高的水产品中。适量的生物胺对于维持人体正常的生理功能是必要的,但是过量的生物胺摄入体内会导致食物中毒、过敏等一系列毒副作用,对人体健康造成极大危害。目前,食品中生物胺含量也是食品品质是否优良的重要指标。因此,对水产品中生物胺含量的测定受到越来越多的重视。近些年来对于生物胺的化学、毒理学以及分析检测技术的研究日趋增加。本文将介绍目前水产品中常见生物胺含量测定的色谱、光谱检测方法以及生物化学检测方法的研究进展,以期为水产品的质量安全提供理论保证。同时对水产品中生物胺检测技术的发展和趋势进行展望。     

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.     

Application of LC with Evaporative Light Scattering Detector for Biogenic Amines Determination in Fair Trade Cocoa-Based Products

In this study, the separation of eight biogenic amines (cadaverine, serotonin, histamine, spermidine, spermine, tyramine, putrescine and β-phenylethylamine) by a liquid chromatography (LC) method with evaporative light scattering detection (ELSD) was performed. The LC–ELSD method was validated by comparison of the results with those obtained through LC–ultraviolet (UV) determination, based on a pre-column dansyl chloride derivatisation step, and the recorded data showed as both analytical methods can be interchangeably used for biogenic amines determination. LC–ELSD methodology showed good precision and permitted to achieve, for standard solutions, limits of detection (LOD) ranging from 0.01 to 0.02 μg ml^?1 and limits of quantitation (LOQ) ranging from 0.03 to 0.05 μg ml^?1. The whole methodology, comprehensive of the homogenization–extraction process and LC–ELSD analysis, has been applied in the analysis of several samples of fair trade cocoa derivatives. The most abundant amine found was histamine for a total amount of biogenic amines in the range 5.81–38.82 μg g^?1. The highest amounts of biogenic amines (BAs) were found in the most processed products but never representing a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable.     

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.     

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).     

Quality and safety of irradiated food regarding biogenic amines: Ras cheese

Food safety and quality became very important, especially with the challenge to ensure safe and healthy foods in regard to chemical hazards. So, this study was conducted to evaluate the quality and safety of irradiated Ras cheese during the storage period, with respect to biogenic amines (BAs). Ras cheese was manufactured, ripened and irradiated by γ‐irradiation at 0, 5, 10 and 15 kGy. The samples were stored in refrigerator at 5 ± 1 °C from where samples were withdrawn at 0, 2, 4 and 6 months for analysis. The results revealed that most sensory scores and chemical properties showed insignificant differences (P ≤ 0.05). The microbial counts were reduced with different degrees according to both storage period and irradiation dose. Also, irradiation was effective in reducing the content of BAs without harming the chemical properties of Ras cheese. The total content of BAs reflects the safety of irradiated Ras cheese and also indicates a high‐quality product in comparison with nonirradiated samples.     

Detection of ten biogenic amines in Chinese commercial soybean paste by HPLC

This paper aimed to develop a high-performance liquid chromatography (HPLC) method that can simultaneously detect ten biogenic amines (BAs) in commercially available soybean paste and to determine the contents of BAs in these products. Heptylamine (Hep) was used as the internal standard reagent and benzoyl chloride as the sample derivatization reagent; Inertsil ODS-3 chromatographic column was set as the stationary phase while the acetonitrile and 5 mmol/L ammonium acetate solution as the mobile phases; the samples were treated with gradient elution and detected at 254 nm (UV). Perchloric acid solution (0.4 mol/L) was employed as the sample extraction solvent to determine the contents of ten BAs in soybean paste products. All BAs were separated within 20 min. The methodological validation suggested good linearity, whereas the linear correlation coefficient of all standards was greater than 0.99 with the linear range of 0.1–80 μg/g, detection limits of 0.01–0.03 μg/g, and quantitation limits of 0.03–0.10 μg/g. For 17 commercially available soybean paste products involved in our study, different samples were distinguished from each other not only by the dramatic differences in the contents of BAs but also by the variance in BA types. Of these BAs, tyramine (Tyr) and agmatine (Agm) were found to be the highest contents in soybean paste products; the contents of histamine (His), 2-phenylethylamine (Phe), serotonin (Ser), spermidine (Spd), and putrescine (Put) were lower, followed by cadaverine (Cad), tryptamine (Try), and spermine (Spm). This method proved to be highly sensitive and effective in determining the contents of BAs in soybean paste products; there was a dramatic difference in the contents of ten BAs among different soybean paste products.     

Biogenic amines assessment during different stages of the canning process of skipjack tuna (Katsuwonus pelamis)

The present research focused on the biogenic amines (BAs) formation in skipjack tuna (Katsuwonus pelamis) throughout the whole canning process. In agreement with its wide employment on this species, on‐board brine immersion freezing (BIF) was tested as post‐mortem processing. The study included fish samples corresponding to different stages of the canning process such as frozen, thawed, cooked and canned; after cooking, two kinds of tuna muscles were considered, that is, whole fillets (main product) and grated muscle (off‐product arising from small pieces). For the BAs (tryptamine, putrescine, cadaverine, histamine, spermidine and spermine) assessment, an HPLC‐DAD method was developed and validated in skipjack tuna samples, in agreement with different parameters such as suitability, linearity, limits of detection and quantification, precision, accuracy and robustness. Tuna submitted on‐board to BIF procedure provided low levels of spermine and spermidine (up to 27.6 mg kg^?1), while contents on the remaining BAs maintained below the limit of detection. Throughout the different stages of the canning process, skipjack tuna showed a low formation of most BAs; interestingly, histamine content was found below 10.6 mg kg^?1 level. The highest values were obtained for spermidine, these related to cooked grated tuna (from 22.6 to 66.7 mg kg^?1) and canned grated tuna (from 70.6 to 104.4 mg kg^?1). Values for pH assessment in all kinds of tuna samples corroborated the results obtained for BAs determination. BIF procedure proved to be an amenable post‐mortem processing to guarantee the quality of canned skipjack tuna.     

A review of the liquid chromatographic methods for the determination of biogenic amines in foods

Biogenic amines (BAs) are biologically active molecules which have aliphatic (putrescine, cadaverine, spermine, spermidine), aromatic (tyramine, phenylethylamine) or heterocyclic (histamine, tryptamine) structures. They can be detected in raw and processed foods which are formed and degraded through several pathways during the metabolic processes of animals, plants and microorganisms. The identification and quantitation procedures of BAs in food samples are very important, because BAs are considered as the indicators of food quality and freshness. The determination of BAs are commonly achieved by separation techniques such as high-performance liquid chromatography (HPLC), gas chromatography (GC) and capillary electrophoresis (CE). In this article, analysis of BAs in foods were reviewed from 2007 to present.     

Effects of rosemary and sage tea extract on biogenic amines formation of sardine (Sardina pilchardus) fillets

Natural antimicrobials and antioxidants from rosemary (Rosmarinus officinalis) and sage tea (Salvia officinalis) were produced using solvent extraction method. The effect of two extracts on ammonia (AMN) and biogenic amines (BAs) formation in vacuum packed sardine (Sardina pilchardus) fillets stored at 3 ± 1 °C was investigated for 20 days. Although the effect of extracts was dependent on specific amine and storage time, phenolic compounds from rosemary and sage tea generally resulted in lower AMN and BAs accumulation in sardine muscle. Putrescine (PUT) and cadaverine (CAD) were the most abundant amines, while histamine (HIS) concentration ranged from 2.05 to 28.77 mg 100g^?1. Rosemary and sage tea extracts significantly reduced HIS, PUT, CAD and trimethylamine accumulation in the fish muscle (P < 0.05) while stimulating effect of extracts was observed on serotonin and agmatine formation. At the end of the storage period, PUT and CAD contents of control were 100‐fold higher than those of treated groups.     

Physicochemical and microbiological changes in postmortem crayfish (Procambarus clarkii) stored at 4 °C and 25 °C

The physicochemical and microbiological parameters of postmortem crayfish stored at 4 °C and 25 °C were evaluated in order to reduce safety risks of crayfish from temperature abuse during transportation and storage. Results showed that hepatopancreas of postmortem crayfish had significantly higher contents of total volatile basic nitrogen, biogenic amines (BAs) and higher microbial loads than tail meat, regardless of storage temperature. Enterobacteriaceae and Pseudomonas counts reached log 6.41 log₁₀ CFU g⁻¹ and 6.31 log₁₀ CFU g⁻¹ in hepatopancreas of crayfish at 25 ℃ for 6 h. Putrescine and cadaverine were the main BAs in tail meat with levels of 28.18 ± 0.73 mg kg⁻¹ and 187.32 ± 3.57 mg kg⁻¹, respectively, whilst cadaverine, spermidine and spermine were the major BAs in hepatopancreas, reaching 283.45 ± 3.95 mg kg⁻¹, 62.87 ± 9.02 mg kg⁻¹ and 155.31 ± 4.55 mg kg⁻¹, respectively, after postmortem storage at 25 °C for 12 h. With time increasing, Acinetobacter, Flavobacterium, Aeromonas and Chryseobacterium at 25 °C and Acinetobacter, Flavobacterium, Psychrobacter at 4 °C in tail meat as well as Bacteroides and Muribaculaceae at 25 °C, and Acinetobacter, Psychrobacter at 4 °C in hepatopancreas, gradually became the major genus at the end of storage. Based on the results of spoilage microorganisms and biogenic amines, crayfish stored at 4 °C and 25 °C could be edible within 24 h and 6 h, respectively.     

Occurrence of biogenic amines in beers produced with malted organic Emmer wheat (Triticum dicoccum)

Because several groups of microorganisms are able to decarboxylate amino acids, the presence of biogenic amines (BA) can be seen as an index of the microbiological quality of the brewing process. BAs were quantified for the first time in the intermediate products and craft beers produced with malted organic Emmer wheat (Triticum dicoccum) in a small size brewery in order to assess the possible presence of critical control points related to biological hazard in the brewing process. BA levels in beers produced exclusively from malted organic Emmer wheat were between 15.4 and 25.2 mg l^–1 in the samples of light beer (Lt) and between 8.9 and 15.3 mg l^–1 in double malt beers (DM) ready for consumption (the beers stored for 90 days at 1–2°C). Cadaverine and tyramine were the main BAs in the Lt and DM beers, respectively. Increased concentrations of BAs seemed to be more related to the heat treatment of the processing product during mashing and wort boiling, rather than to the fermentation process. Much lower concentrations were found in finished beers obtained from 50% malted organic Emmer wheat and 50% malted barley (up to 3.2 mg l^–1) or from 30% malted Emmer wheat (up to 8.3 mg l^–1). Thus, Emmer wheat malt can be a useful alternative to wheat and spelt for the production of beer with a limited content of BA, if the processing technology is kept under control.     

Determination and distribution of biogenic amines in bee pollen

The natural decomposition of proteins yields peptides and amino acids, which are susceptible to further decay, resulting in formation of biogenic amines (BAs) in proteinaceous foods such as bee pollen (BP). However, excessive consumption of BAs will bring side effects, so it is vital to study the presence of BAs in BP. In this study, a sensitive and reliable ultra ‐ high‐performance liquid chromatography–tandem mass spectrometric method was used for the determination of 12 BAs in BP. Large variations in the distribution and levels of BAs were observed with its total concentrations ranging from 6837.4 to 25444.2 μg kg^?1. In addition, the changes in the levels of BAs during storage and fermentation were preliminarily investigated. The results showed that fermentation significantly altered the quantities of BAs in rape BP, while changes during storage were modest. The results of this study could be applied for quality evaluation and risk assessment of BP in near future.