Determination Of Biogenic Amines Research Articles

A new method for the determination of biogenic amines in cheese by LC with evaporative light scattering detector

This paper presents a new LC method with evaporative light scattering detection (ELSD), for the separation and determination of the biogenic amines (histamine, spermidine, spermine, tyramine, putrescine and β-phenylethylamine) which are commonly present in cheese, as their presence and relative amounts give useful information about freshness, level of maturing, quality of storage and cheese authentication. The LC–ELSD method is validated by comparison of the results with those obtained through LC–UV determination, based on a pre-column dansyl chloride derivatisation step. The obtained data demonstrate that both methods can be interchangeably used for biogenic amines determination in cheese. The new LC–ELSD method shows good precision and permits to achieve, for standard solutions, limit of detection (LOD) values ranging from 1.4 to 3.6mgL−1 and limit of quantitation (LOQ) values ranging from 3.6 to 9.3mgL−1. The whole methodology, comprehensive of the homogenization–extraction process and LC–ELSD analysis, has been applied in the analysis of a typical Calabria (Southern Italy) POD cheese, known as Caciocavallo Silano. The most aboundant amine found was histamine, followed, in decreasing order, by tyramine, spermine, putrescine, β-phenylethylamine and spermidine, for a total amount of 127mgkg−1. This value does not represent a possible risk for consumer health, according to the toxicity levels reported in literature and regarded as acceptable.

Chromatographic determination of biogenic amines in wines after treatment with ionic liquids as novel media

Room temperature ionic liquids (RTIL), 1-butyl-3-methylimidazolium hexafluorophosphate ([C(4) MIM][PF(6) ]), were used as the novel media for derivatization, extraction and preconcentration of biogenic amines (BAs) in wines. Six BAs, tryptamine (Try), phenylethylamine (Phe), putrescine (Put), cadaverine (Cad), tyramine (Tyr) and spermine (Spe) were selected as the model compounds and dansyl chloride (Dns-Cl) as the derivatizing reagent. The derivatizations of amines were conducted in water-IL two-phase system, in which the partition property of related substance in the IL was investigated and the mechanism (http://www.iciba.com/mechanism/) of derivation and extraction of amines was discussed. The influencing factors, including sample volume, derivatizing reagent concentration, pH value and ultrasound reaction time, were optimized. The Dns-amines were separated by high-performance liquid chromatography (HPLC), and detected with UVD at 254 nm. For each of the tested amines, a good linearity was obtained in the concentration range of 0.1-20 mg/L with the correlation coefficient (R) of 0.9814-0.9930. The limits of detection reached μg/L level and the relative standard deviations (RSD, n=3) were between 3.2 and 8.1%. Satisfactory recovery for each BA was obtained, ranging from 82.3 to 114.0%. The developed method was successfully applied to determine six BAs in red wines and Chinese yellow wine.

Biogenic Amines in Wines and Pomegranate Molasses—A Non-Ionic Micellar Electrokinetic Chromatography Assay with Laser-Induced Fluorescence Detection

Biogenic amines in wine samples and pomegranate molasses were detected and quantified by a nonionic micellar electrokinetic chromatography method coupled to laser-induced fluorescence detection. The method provides a satisfactory and fast separation of seven biogenic amines and matrix peaks in food samples in less than 9 min. Detection limits are between 0.42 and 1.26 nM, and precisions were lower than 3% RSD for peak areas. The recovery values were between 93% and 104%, depending on the food matrix. The method is sensitive and rapid and widely applicable for the determination of biogenic amines in wine samples and fruit molasses.

Determination of biogenic amines in lake water by micellar electrokinetic chromatography with fluorescence detection after derivatization with fluorescamine

A simple and rapid method has been developed for the determination of biogenic amines in lake water using micellar electrokinetic chromatography with fluorescence detection. Separation of fluorescamine derivatized biogenic amines was accomplished by using borate buffer of pH 9.5 containing 40 mM of sodium dodecyl sulphate. The method has been optimized with respect to fluorescamine concentration, reaction pH, reaction time, separation voltage and injection time. Detection was performed by using UG-11 excitation filter and 495 nm emission filter. The proposed method for histamine, tyramine and dopamine allowed their separation within 2 min with detection limits in nM range. The interday and intraday reproducibility of peak areas were less than 6.5%. Recovery of spiked samples was 95.76–116.31%.

Determination of biogenic amines in Cheonggukjang 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.

Development of an ionic liquid-based ultrasonic-assisted liquid–liquid microextraction method for sensitive determination of biogenic amines: Application to the analysis of octopamine, tyramine and phenethylamine in beer samples

A simple and efficient method, ionic liquid-based ultrasound-assisted liquid–liquid microextraction, has been developed for the determination of three biogenic amines including octopamine (OCT), tyramine (TYR) and phenethylamine (PHE). Fluorescence probe 2, 6-dimethyl-4-quinolinecarboxylic acid N-hydroxysuccinimide ester was applied for derivatization of biogenic amines and high-performance liquid chromatography coupled with fluorescence detection was used for the determination of the derivatives. The factors affecting the extraction efficiency, such as the type and volume of ionic liquid, ultrasonication time and centrifugation time have been investigated in detail. Under the optimum conditions, linearity of the method was observed in the range of 0.5–50 μg mL −1 for OCT and TYR, and 0.025–2.5 μg mL −1 for PHE, respectively, with correlation coefficients ( γ) > 0.996. The limits of detection ranged from 0.25–50 ng mL −1 (S/N = 3). The spiked recoveries of three target compounds in beer samples were in the range of 90.2–114%. As a result, this method has been successfully applied for the sensitive determination of OCT, TYR and PHE in beer samples.

LC Determination of Trace Biogenic Amines in Foods Samples with Fluorescence Detection and MS Identification

A pre-column derivatization method for the simple, sensitive determination of biogenic amines using 10-ethyl-acridine-3-sulfonyl chloride (EASC) as labeling reagent with fluorescence detection and mass spectrometry (MS) identification has been developed. After pre-column derivatization, the labeled biogenic amines were separated on a Hypersil BDS-C18 column by gradient elution. The derivatives showed an intense protonated molecular ion corresponding m/z [M + H]+ in positive-ion mode. The collision-induced dissociation of protonated molecular ion formed specific fragment ions at m/z 196.5, m/z 222.7, m/z 224.4 and m/z 272.5, m/z 286.2. Satisfactory linear responses were observed at the concentration range of 0.02–10 μmol L−1 with coefficients of >0.9993. Detection limits obtained by the analysis of a derivatized standard containing 0.2 pmol of each biogenic amine, were from 20.22 to 109.2 fmol (at a signal-to-noise ratio of 3). The relative standard deviations of retention times and peak areas for each biogenic amine were <0.96 and 3.22%, respectively. Recoveries except for PUT were in the range of 96.7–103.6% for chicken sausage and 95.8–104.6% for pork sausage The established method for the determination of biogenic amines except for PUT from real samples was satisfactory.

High-throughput sensing microtiter plate for determination of biogenic amines in seafood using fluorescence or eye-vision

A new optical sensing microplate was developed for rapid screening for the presence of biogenic amines (BAs) in seafood samples with high sensitivity. The deposition of a sensing spot (containing a chameleon dye (Py-1) in a polymeric cocktail) on the bottom of the wells of a standard microplate renders the plate a new sensing tool for a rapid and parallel detection of up to 96 (real) samples. This sensing microplate enables (1) a semi-quantitative readout of analyte concentration by eye-vision, (2) a rapid fluorescence readout of 96 samples with standard instrumentation in less than two minutes (unlike chromatographic and electrophoretic methods), (3) a statistically robust data evaluation (with 8-12 replicates) and (4) a rapid parallel sample preparation with standard 8 or 12-channel micropipettes. On reaction with biogenic amines, the dye shows a significant visible color change from blue over green to red color. The appearance of red color favorably coincides with the concentration of BAs that can induce symptoms of poisoning. The linear ranges of fluorescence calibration data for six biogenic amines cover the clinical toxicological relevant range of BAs that is too low to be detected by the human nose. The LODs range from 0.16 to 0.56 μg mL(-1), with correlation coefficients (r(2)) between 0.985 and 0.999. Finally, the evolution of spoilage of four fish samples (monitored by determination of their BA status) and the increase of their total amine content were found to agree well with previous data on time-dependent evolution of BAs in fish.

Determination of Biogenic Amines and Endotoxin in Squid, Musky Octopus, Norway Lobster, and Mussel Stored at Room Temperature

Little research has been published on the indicators of spoilage in Mediterranean molluscan shellfish and crustaceans. Thus is why we studied changes in the concentrations of endotoxin and four biogenic amines (histamine, putrescine, tyramine and cadaverine) in European common squid (Loligo subulata, Lamarck, 1798), musky octopus (Eledone moschata, Lamarck, 1798), Norway lobster (Nephrops norvegicus, Linnaeus, 1758), and mussel (Mytilus galloprovincialis, Lamarck, 1819) from the Adriatic Sea stored at room temperature for 24 h. Endotoxin load in fresh squid, Norway lobster, and mussel (<1 EU mg-1) indicated good microbiological quality of raw samples. Biogenic amine index (as the sum of histamine, putrescine, tyramine, and cadaverine) correlated well with endotoxin load in squid (r=0.978, p<0.001) and musky octopus (r=0.874, p<0.01). A good correlation was also found between endotoxin and putrescine in Norway lobster (r=0.777, p<0.05). The highest endotoxin load was found in decomposed mussels and was associated with histamine alone. In conclusion, increase in biogenic amine levels is species-specific. Endotoxin analysis could be used for rapid assessment of microbiological quality of cephalopods and shellfish.

THE POTENTIAL OF CHRYSEOBACTERIUM SPECIES TO PRODUCE BIOGENIC AMINES

ABSTRACTThe food spoilage potential of the genus Chryseobacterium has not been studied in equal detail than the taxonomy of this genus. The ability of seven Chryseobacterium species to produce biogenic amines (BAs) at different temperatures and sodium chloride concentrations, was investigated by using a modified Niven medium. Temperatures at and below 15C and 25C seemed to have a definite inhibitory effect on the production of BAs in some Chryseobacterium species. Salt concentrations in excess of 4% would be needed to prevent amine production of Chryseobacterium species in food products. Chryseobacterium species have therefore the ability to decarboxylate some precursors of BAs, making them important spoilage bacteria in dairy food products.PRACTICAL APPLICATIONSThe determination of biogenic amines (BAs) is important not only from the point of view of their toxicity (symptoms such as headache, fever and vomiting may occur) but also because they can be used as indicators of the degree of freshness or spoilage of food. The ability of Chryseobacterium spp. to produce BAs, at low temperatures, is of great significance to the dairy and other food industries, because of the typical off‐flavors and putrid odors that are produced. Refrigerated products could cause amine poisoning if they were contaminated with amine‐producing bacteria, e.g., Chryseobacterium spp.. A previous study found that the content of BAs increases even during storage at +4C. It must be taken note of that most amines are heat‐stable and only partially destroyed in 3 h at 102C or 90 min at 116C. These findings stress the importance of the hygienic production and handling of food products and maintenance of the cold chain throughout production and distribution.

Determination of Biogenic Amines in Herby Cheese

Herby cheese (Otlu peynir) is widely produced and consumed in eastern parts of Turkey, and is generally made from sheep milk. The biogenic amines and organic acids content, microbiological, and chemical properties of Herby cheese were determined on 30 samples obtained from retail markets in East Anatolia Region, Turkey. The most important biogenic amines were tyramine (range 18.0–1125.5 mg kg−1), followed by cadaverine varied from not detected to 1844.5 mg kg−1. Histamine content generally was found higher than 100 mg kg−1. The concentration of amines in some cheeses was much higher than the toxic dose limits. Generally, total amine content was found high in samples that had high nitrogen fraction and organic acids value. Levels of biogenic amines were significantly correlated with the organic acids of Herby cheese. The basic organic acid was found the lactic acid ranging from 585.4 to 26480.3 mg kg−1, followed by butyric acid varying between 314.8 and 7329.6 mg kg−1 values. Lipolysis and nitrogen fraction were determined high in ripened Herby cheese samples. Coliform counts of samples showed lower than 100 cfu g−1 in the samples. Significant (p < 0.01) positive correlation was observed between acetic acid and phenylethylamine, putrescine, cadaverin, histamine, and tyramine.

Rapid automated high performance liquid chromatography method for simultaneous determination of amino acids and biogenic amines in wine, fruit and honey

This paper reports a new, simple, rapid and economical method for routine determination of 24 amino acids and biogenic amines in grapes and wine. No sample clean-up is required and total run time including column re-equilibration is less than 40 min. Following automated in-loop automated pre-column derivatisation with an o-phthaldialdehyde, N-acetyl- l-cysteine reagent, compounds were separated on a 3 mm × 25 cm C 18 column using a binary mobile phase. The method was validated in the range 0.25–10 mg/l; repeatability was less than 3% RSD and the intermediate precision ranged from 2 to 7% RSD. The method was shown to be linear by the ‘lack of fit’ test and the accuracy was between 97 and 101%. The LLOQ varied between 10 μg/l for aspartic and glutamic acids, ethanolamine and GABA, and 100 μg/l for tyrosine, phenylalanine, putrescine and cadaverine. The method was applied to grapes, white wine, red wine, honey and three species of physalis fruit. Grapes and physalis fruit were crushed, sieved, centrifuged and diluted 1/20 and 1/100, respectively, for analysis; wines and honeys were simply diluted 10-fold. It was shown using this method that the amino acid content of grapes was strongly correlated with berry volume, moderately correlated with sugar concentration and inversely correlated with total acidity.

Use of small diameter column particles to enhance HPLC determination of histamine and other biogenic amines in seafood

Pre-column and post-column HPLC derivatization methods were modified and evaluated for the identification and quantification of nine biogenic amines in seafood. Two HPLC methods with column particles of 1.8 μm or 3 μm in diameter were modified and compared to classical methods using 5 μm column particles. Both pre-column derivatization with dansyl chloride and post-column derivatization with O-phthalaldehyde were studied. The HPLC methods were compared with respect to the time of elution, eluent consumption, backpressure as well as separation, sensitivity, recovery and repeatability for determination of biogenic amines in lean canned tuna and fatty frozen herring. The modified methods using smaller column particles of 1.8 μm or 3 μm allowed biogenic amines to be separated and quantified faster (23–59%) and with less eluent consumption (59–62%) than classical HPLC methods. Backpressures were below 170 bar and this allowed the use of classical HPLC systems rather than dedicated and costly ultra-high-pressure liquid chromatography (UHPLC) equipment. Biogenic amine separation, sensitivity, recovery and repeatability for the modified methods were similar to or performed better than for the classical HPLC methods. Due to reduced analyses time and eluent consumption the modified HPLC methods would improve biogenic amine analysis.

A 4-hydroxy- N′-[( E)-(2-hydroxyphenyl)methylidene]benzohydrazide-based sorbent material for the extraction-HPLC determination of biogenic amines in food samples

A sorbent material based on a newly synthesized hydrazone ligand, 4-hydroxy- N′-[( E)-(2-hydroxyphenyl)methylidene]benzohydrazide was prepared by immobilizing the ligand into a silica sol–gel matrix. The capability of the sorbent material for the extraction of seven biogenic amines (BAs), i.e., tryptamine (TRY), β-phenylethylamine (PEA), putrescine (PUT), cadaverine (CAD), histamine (HIS), tyramine (TYR), and spermidine (SPD) was studied. Under the adopted conditions, the sorbent showed good selectivity towards PUT, CAD, HIS and SPD (% extraction (%E) > 96) while %E for TYR, TRY and PEA were 82.0, 78.9 and 46.4%, respectively. The sorbent could be used up to six extraction cycles for SPD, CAD and PUT and was applied to the determination of food samples (“budu”, ketchup, orange juice, soy sauce) that were spiked with 20 mg L −1 of the BAs. The extracted analytes were derivatized with dansyl chloride before the HPLC determination. With the exception of HIS and TYR in “budu” sample, reasonable recoveries were found for the other analytes in all the tested food samples.

Non‐ionic micellar electrokinetic chromatography with laser‐induced fluorescence: A new method tested with biogenic amines in brined and dry‐salted fish

A non-ionic micellar electrokinetic chromatography method coupled to laser-induced fluorescence detection was developed for the quantitative determination of biogenic amines. Complete resolution of six biogenic amines - FITC derivatives - was achieved in less than 10 min, employing 10 mM Brij 35 in 75 mM borate buffer and at pH 9.7 as the running electrolyte. Relative fluorescence intensities of biogenic amines enhanched considerably and separation time decreased considerably when Brij 35 was substituted for SDS in the same buffer. The detection limits of the method for biogenic amines were found between 0.416 and 1.26 nM. The precisions for the corrected peak areas were calculated as between 1.63 and 3.24 %RSD. The applicability of the method was demonstrated by analyzing biogenic amines in the processed, brined, and dry-salted fish samples. The method is simple and rapid, and widely applicable for the determination of biogenic amines in food samples.

Effect of grape storage time on biogenic amines content in must

The presence of amines in wine can produce a negative impact on the aroma of the product and, in some cases, detrimental health effects. Several factors affect the content of amines in wine, among those, the presence of biogenic amines in must. The aim of this work was to value the effect of three storage times of grapes (0, 24 and 48 h), in the winery, and the effect on the concentration of biogenic amines in musts derived from three white grape cultivars Chardonnay, Riesling and Sauvignon blanc. The determination of biogenic amines in must was carried out by HPLC with fluorimetric detector. In musts from sterilised grapes, only ethanolamine, ethylamine, putrescine, and tyramine (only for Chardonnay) were found; besides these abovementioned amines, in musts from non-sterilised grapes which were stored in the winery before crushing, cadaverine, tyramine and agmatine were also found. The individual and total amine contents of the must are significantly influenced by the time of grape storage in the winery.

Disposable biosensors for determination of biogenic amines

This work reports monoamine oxidase (MAO)/horseradish peroxidase (HRP) and diamine oxidase (DAO)/horseradish peroxidase (HRP) based biosensors using screen-printed carbon electrodes for the determination of biogenic amines (BA). The enzymes have been covalently immobilized onto the carbon working electrode, previously modified by an aryl diazonium salt, using hydroxysuccinimide and carbodiimide. The detection has been performed by measuring the cathodic current due to the reduction of the mediator hydroxymethylferrocene at a low potential, 250 mV vs screen-printed Ag/AgCl reference electrode. The experimental conditions for the enzymes immobilization, as well as for the main variables that can influence the chronoamperometric current have been optimized by the experimental design methodology. Under these optimum conditions, the disposable biosensors have been characterized. A linear response range from 0.2 up to 1.6 μM and from 0.4 to 2.4 μM of histamine was obtained for DAO/HRP and MAO/HRP based biosensors, respectively. The biosensor construction was highly reproducible, yielding relative standard deviations of 10% and 11% in terms of sensitivity for DAO/HRP and MAO/HRP based biosensors, respectively. The capability of detection, 0.18 ± 0.01 μM in the case of DAO/HRP and 0.40 ± 0.04 μM ( α = 0.05 and β = 0.005) for MAO/HRP based biosensors, and the biosensor sensitivity towards different BA has also been analyzed. Finally, the developed biosensors have been applied to the determination of the total amine content in fish samples.

Determination of biogenic amines in Korean traditional fermented soybean paste (Doenjang)

Biogenic amines, produced by bacterial decarboxylation of amino acids, have been associated with toxicological symptoms in food products. Twenty-three samples of traditionally available Korean fermented soybean paste samples (Doenjang) were analyzed in order to determine the content of biogenic amines. Amines were extracted with 0.4 M perchloric acid and derivatized with dansyl chloride. Nine biogenic amines were separated from Doenjang samples by high performance liquid chromatography using gradient elution (acetonitrile and ammonium acetate), and detected with spectrophotometric UV–vis detection at 254 nm. The pH value of all the samples was ranged from 4.8 to 6.0, and the strong amino acid decarboxylase activity was found to be in an acidic environment. The mean values of biogenic amines (tryptamine, 2-phenyl–ethylamine, putrescine, cadaverine, agmatine, histamine, tyramine, spermidine and spermine) determined in 23 Doenjang samples were found to be 18.37, 82.03, 70.84, 34.24, 47.32, 26.79, 126.66, 74.41 and 244.36 mg%, respectively. The findings of this study enhance the safety of not only Doenjang but other salted and/or fermented food products.

Sensitive profiling of biogenic amines in urine using CE with transient isotachophoretic preconcentration

A transient ITP-CZE system is proposed for the determination of biogenic amines in urine. The complete separation and identification of dopamine, tyramine (TA), tryptamine (T), serotonin, epinephrine, norepinephrine, and normetanephrine have been achieved in a twofold diluted urine sample (in which the analytes were below the LODs by normal CZE). The tITP preconcentration conditions were created by introducing a 30 mM solution of NaOH, containing 0.1% hydroxypropylcellulose (pH 6.5 adjusted with MES), and 0.1 M HCl before and after the sample zone to work as leading and terminating electrolytes, respectively. This allowed the LODs of direct UV absorption detection to be decreased down to the 10(-8) M level that is comparable with the sensitivity thresholds of LIF detection or inline SPE-CE. The RSDs of migration time and peak area for real-biofluid analysis were in the range of 0.1-4.5% and 0.8-16% (n=3), respectively. Quantification of dopamine, TA, T, and serotonin was performed using internal calibration. To the best of our knowledge, this is the first report on probing urinal biogenic amines and their metabolites by tITP-CZE.

High-Performance Liquid Chromatography−Ultraviolet Detection Method for the Simultaneous Determination of Typical Biogenic Amines and Precursor Amino Acids. Applications in Food Chemistry

A reversed-phase high-performance liquid chromatography (HPLC) method was developed for the simultaneous determination in food of biogenic amines and their precursor amino acids after a precolumn derivatization with dansyl chloride. The chromatographic conditions, selected to be suitable for mass spectrometry detection, were optimized through experimental design and artificial neural networks. The HPLC-UV method was validated by comparing the separation results with those obtained through a HPLC method, working under the same chromatographic conditions but employing mass spectrometry detection. The HPLC-UV method was then applied to the analysis of different food samples, namely, cheese, clams, salami, and beer. For all of the matrices, recoveries (relative standard deviation always <5%) always >92% were obtained. The results are discussed as a function of the total biogenic amine content and of the concentration ratio between amines and precursor amino acids.