Biogenic Amines In Beer Research Articles

Ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction of trace biogenic amines in fish and beer based on solidification of floating organic droplet.

It is important to detect the presence of biogenic amines (BAs) as indicators of food freshness. The purpose of this study was to develop a novel ultrasonic-microwave synergistic supramolecular solvent liquid-liquid microextraction based on solidification of floating organic droplet (UMS-SUPRAS-SFO-LLME) combined with high-performance liquid chromatography for the determination of BAs. The physical properties and microstructure of SUPRAS based on 1-dodecanol and tetrahydrofuran were studied, and the extraction conditions such as the SUPRAS volume, the UMS process, and the centrifugal conditions were optimized. The results for the extraction kinetics and thermodynamics showed that UMS-SUPRAS-SFO-LLME is a spontaneous, endothermic diffusion process. The linear ranges of this method are 0.1-2.0×105 ng·mL-1 (R2>0.994), the limits of detection are 4.0×10-3-6.0×10-2 ng·mL-1, and the recoveries were 96.28-103.15%. Compared with existing analysis methods, UMS-SUPRAS-SFO-LLME is a sensitive, green and economical sample pretreatment method for analyzing the enrichment of BAs in beer and fish.

The Measurement of Hazardous Biogenic Amines in Non-Alcoholic Beers: Efficient and Applicable Miniaturized Electro-Membrane Extraction Joined to Gas Chromatography-Mass Spectrometry.

The determination of biogenic amines (BAs) as serious food contaminants and chemical indicators of unwanted microbial contamination or deficient processing conditions in non-alcoholic beers is of great interest for the beverage industries. In the present investigation, the combination of hollow fiber-electro-membrane extraction (HF-EME) and dispersive liquid-liquid microextraction (DLLME) followed by gas chromatography-mass spectrometry (GC/MS) was applied for the analysis of histamine, putrescine, tyramine, cadaverine in non-alcoholic beers. EME is fundamentally based on the electrostatic attraction, diffusion and solvability of analytes in a selected acceptor phase. This membrane-based extraction technique promoted selectivity and the enrichment factor. The DLLME process reduced the volumes of organic solvents and make the coupling of HF-EME to the CG/MS conceivable. The leading variables, which have a great effect on extraction recovery, were optimized. The relative standard deviation was achieved between 4.9 and 7.0%. The recoveries were between 94% and 98%. The limit of detection and limit of quantification were found to be 0.92-0.98 ng mL-1 and 3.03-3.23 ng mL-1, respectively. The enrichment factor was calculated in the range 36-41. The achievements revealed that putrescine and tyramine, with concentrations of 3.87 and 2.33 µg g-1, were at the highest concentration in non-alcoholic beers. This offered method with great benefits could help beverage industries to monitor the concentration of BAs in beers and control them.

Occurrence and Levels of Biogenic Amines in Beers Produced by Different Methods.

The concentration of biogenic amines (BAs) in beer depends, among other factors, on the activity of microorganisms, in particular lactic acid bacteria. In this work an analytical method based on derivatization with tosyl chloride and high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was used to determine 17 BAs in samples of commercially available beers, and to monitor the changes in concentration of several BAs throughout the fermentation process. In some of the analysed samples the concentration of BAs exceeded the safety threshold for consumers. During the fermentation stage of home-brewing of ale the concentration of spermine in the wort increased until the end of the stormy fermentation, to then drop below the initial concentration at the end of fermentation, and below the LOQ after refermentation. The results of the study indicate that monitoring the total content of BAs is required due to the potential risk to human health.

Solid phase “on-situ” quadraplex isotope dimethyl labeling for the analysis of biogenic amines in beers by liquid chromatography-high resolution mass spectrometry

A simple magnetization method was developed for the preparation of magnetic materials from conventional solid phase packing though coprecipitation and solvothermal approaches. And the prepared magnetic materials were used for magnetic solid phase extraction (MSPE) of biogenic amines (BAs) from beers. Furthermore, to improve the analytical throughput, a solid phase “on-situ” quadraplex isotope dimethyl labeling method was developed for the quantification of BAs by liquid chromatography-high resolution mass spectrometry (LCHRMS). Compared to conventional in-solution labeling, the “on-situ” labeling could simplify the sample preparation procedure and efficiently remove the residuals such as inorganic salts and excessive labeling reagents. The quadraplex labeling, which enabled three real samples and one internal standard sample to be analyzed simultaneously in a single LCHRMS run. For the tested 8 BAs (cadaverine, phenethylamine, spermine, spermidine, tyramine, histamine, putrescine and tryptamine), LODs of 0.02–0.05 μg/L and LOQs of 0.05–0.1 μg/L were achieved at good reproducibility (RSD of 0.5–4.6% and 2.2–7.0% for intra- and inter-day reproducibility, respectively). With this method, six beer samples were analyzed, and these 8 BAs were all detected in the range of low μg/L to 2.9 mg/L, which were lower than maximal residual level (MRL) required in the regulations of China and EU.

Occurrence of biogenic amines in beers from Chilean market

The presence of tryptamine, phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine was determined for the first time in beers commercialized in Chilean market. Dansylated amines were separated on a Zorbax-XDB C18 column using a binary mobile phase composed of acetonitrile and 20 mmol L−1 ammonium formate, pH 5.5. Chromatographic conditions were optimized using an experimental design, and validation was carried out following ICH recommendations. Calibration data fitted a linear regression model with R2 > 0.996. Repeatability (n = 6) and intermediate precision (n = 3) in matrix showed RSD values lower than 5.00% and 5.21%, respectively. Recoveries at three different concentrations ranged from 75.50 to 96.48%. The proposed method was applied to determine the biogenic amines content in 101 beer samples produced by macro- (n = 65) and microbreweries (n = 36) using low (n = 63) and high fermentations (n = 38). Biogenic amines content in beer samples ranged from 0.53 to 85.04 mg L−1, from which beers produced by microbreweries (19.13 ± 16.92 mg L−1) showed a significant higher biogenic amines content (P = 0.0021) than beers produced by macrobreweries (9.65 ± 4.50 mg L−1). Putrescine was the principal biogenic amine found in all kind of beers regardless origin, and the type of fermentation and brewery. Only 2 samples presented relevant levels of histamine and tyramine, but both below the limits reported by the European Food Safety Authority (EFSA). Thus, it can be concluded that beers commercialized in Chile are not a serious toxicological risk (e.g. hypertensive crisis) regarding the type and content of biogenic amines.

Determination of biogenic amines in beer and wine by capillary electrophoresis–tandem mass spectrometry

A capillary electrophoresis–tandem mass spectrometry (CE–MS/MS) method for the simultaneous assessment of nine biogenic amines (spermine, spermidine, putrescine, cadaverine, histamine, phenylethylamine, tryptamine, tyramine, and urocanic acid) in commercial samples of beer and wine is introduced. The samples were submitted to a simple clean-up step with poly(vinylpolypyrrolidone) followed by filtration. Electrophoretic separation in a polyvinyl alcohol (PVA)-coated capillary using 0.5molL−1 acetic acid (pH 2.5) as background electrolyte and detection by electrospray-tandem mass spectrometry was employed. The range of the correlation coefficients of the calibration curves of the analyzed compounds was 0.996–0.999, and the limits of detection and limits of quantification were in the range of 1–2μgL−1 and 3–8μgL−1, respectively. The recovery values for samples spiked at three concentration levels (0.2, 0.5, and 1.0mgL−1) ranged from 87 to 113% with standard deviation not greater than 5.8%. The use of a PVA-coated silica capillary allows suppressing the electroosmotic flow and, consequently, increasing of the separation efficiency. The method was successfully used to determine biogenic amines in commercial samples of beer and wine.

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.

A novel dispersive liquid–liquid microextraction (DLLME) gas chromatography-mass spectrometry (GC–MS) method for the determination of eighteen biogenic amines in beer

A novel dispersive liquid–liquid microextraction (DLLME) gas chromatography mass-spectrometry (GC–MS) method was developed for the determination of 18 biogenic amines in beers. The method features the simultaneous extraction/derivatization of the amines providing a simple and fast mode of extract enrichment. A mixture of acetonitrile (dispersive solvent; 1.0 mL), toluene (extractive solvent; 325 μL), and isobutyl choloroformate (derivatizing reagent; 25 μL) was used as extractive/derivatizing reagent, added to 5 mL of sample. The proposed method showed good linearity (correlation coefficients > 0.997), good recoveries (from 72 to 113%), and good intra-day precision (below 13%) and inter-day precision (below 14%). Moreover, detection limits were never over 2.9 μg L−1. The developed method was successfully applied to the analysis of 22 beer samples commercialized in Portugal. Fourteen of the eighteen biogenic amines analyzed were found in most of the beers, with predominance of putrescine, tyramine, dimethylamine, cadaverine, pyrrolidine and 1,3-diaminopropane.

Simultaneous determination of 23 amino acids and 7 biogenic amines in fermented food samples by liquid chromatography/quadrupole time-of-flight mass spectrometry

A novel liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-Q-TOFMS) method was developed for the simultaneous determination of 23 amino acids and 7 biogenic amines in food samples. These analytes were pre-column derivatized with dansyl chloride and then separated in an Acquity column (1.7 μm; 2.1 mm × 100 mm). The separation of 31 compounds including an internal standard was achieved within 25 min at a flow rate of 0.2 mL/min. The method linearity for each amino acid and biogenic amine had a relatively wide range with r(2)>0.99. The intra- and inter-day precision, expressed as relative standard deviation (RSD), ranged from 1.1 to 4.6% and from 2.0 to 11.2%, respectively. The limit of detection was between 0.005 and 0.4 μg/mL. With a simple dilution, recoveries of around 80-120% were obtained for most of the compounds. No significant matrix effect was observed, and the developed method was successfully applied to the analysis of amino acids and biogenic amines in beer, cheese and sausage samples.

Determination of biogenic amines in beer with pre-column derivatization by high performance liquid chromatography

Eighteen samples of commercially available Chinese beer were analyzed in order to determine the content of biogenic amines. The method involves pre-column derivatization of the amines with 4-chloro-3,5-dinitrobenzotrifluoride (CNBF) and subsequent analysis by RP-HPLC (reversed phase-high performance liquid chromatography) with diode array detection. The labeled biogenic amines were separated on a Kromasil C18 column (250 mm × 4.6 mm, 5 μm) at room temperature and UV detection was applied at 254 nm. The separation of seven labeled biogenic amines was achieved within 22 min by elution acetonitrile and HAc–NaAc buffers. The method linearity, calculated for each biogenic amine, has a correlation coefficient higher than 0.9925, in concentrations ranging from 2.9 μmol L −1 to 565 μmol L −1. Detection limits of biogenic amines were 0.056–0.87 μmol L −1, at a signal-to-noise ratio of 3. The proposed method has been applied to the quantitative determination of spermine, phenethylamine, spermidine, histamine, tyramine, tryptamine and putrescine in beer with recoveries of 91.9–103.1% and R.S.D. of 2.86–5.63%. Quantitation is relative to external standards. The results showed that each kind of beer examined contained at least three biogenic amines. Putrescine, histamine and tyramine were detected in all samples. Spermidine was detected in 89% of the beers. Spermine, tryptamine and phenylethylamine occurred in 78%, 61% and 44% of the beers examined, respectively. These levels were below the level that may elicit direct adverse reactions for most consumers.

Determination of aliphatic amines by cation‐exchange chromatography with suppressed conductivity detection after solid phase extraction*

A sensitive method for the determination of underivatized aliphatic amines based on cation exchange chromatography coupled with suppressed conductivity detection scheme and solid phase extraction (SPE) procedure has been developed. A surface modified styrene divinylbenzene polymeric sorbent, based on a reversed-phase (RP) and strong cation exchange (SCX) mixed mode was used as an active material for the SPE of amines. The conductometric capabilities of several aliphatic mono- and polyamines, expressed in terms of molar sensitivity (nS/microM), were determined. The LODs, obtained without the SPE treatment, ranged between 20 and 65 nM for putrescine and 2-butylamine, respectively. The resulting calibration plots for the aliphatic amines were generally linear over about three orders of magnitude, with correlation coefficients >0.98. The LODs of amines decreased generally by one factor when SPE procedure, using BaCl(2)/H(2)SO(4) eluents, has been adopted. The proposed SPE procedure, seems to offer good results in terms of preconcentration, recoveries and cleanup of samples. The proposed methodology was successfully tested for the quantitative determination of some biogenic amines in beer and tuna.

Determination of biogenic amines in beers and brewing-process samples by capillary electrophoresis coupled to laser-induced fluorescence detection

We have developed a sensitive capillary electrophoretic method for the simultaneous determination of 10 biogenic amine which usually appears in beer samples, using laser induced-fluorescence. Sample amines were first derivatized and filtered and then separated with an uncoated capillary tubing in the presence of 50 mM sodium borate and 20% acetone at pH 9.3 at 30 kV. It was possible to analyze biogenic amines in brewing-process samples and in beer samples in less than 30 min, obtaining detection limits between 0.3 μg l −1 for ethylamine and 11.9 μg l −1 for 1,6-hexanodiamine.

The influence of the brewing process on the formation of biogenic amines in beers.

Biogenic amines, as dabsyl derivatives, were determined in beer samples, intermediate products, and raw materials (malt and maize) by HPLC. A procedure for the extraction of the amines from malt and maize with diluted hydrochloric acid was optimised by combining a Response Surface Methodology with a Simultaneous Decision Making Approach. The results of the analysis indicate that, in brewing, technology and hygiene are the decisive factors that determine the amine concentrations in the final product.

Biogenic amine profile of the most popular Polish beers

Biogenic amines play an important physiological role in mammals and high amounts of some exogenous amines in the human diet may contribute to a wide variety of toxic effects. These amines are commonly found in many foodstuffs, particularly in fermented products such as cheese, meat products, beer and wine. Therefore the typical level of biogenic amines and some polyamine-derived secondary metabolites in the most popular Polish beers was evaluated. Twenty-seven samples of five types of beer purchased from commercial sources were analysed by reversed-phase high performance liquid chromatography (HPLC) and spectrophotometric detection after post-column derivatization with dansyl chloride. The total average level of biogenic amines in beers was 16.15±2.89 mg/L. The highest concentration of biogenic amines were: spermine 8.43±3.61 mg/L (n=27), spermidine 3.37±2.07 mg/L (n=25) and putrescine 1.75±0.79 mg/L (n=27). Other biogenic amines were determined at concentration less than 2.0 mg/L. There were no essential differences in the total average amine levels among five types of tested beers: lager, pilsner, strong, stout and non-alcoholic. Individual differences in the biogenic amines present and their levels between breweries were pointed out.

A GC-MS method for quantitation of histamine and other biogenic amines in beer

A GC-MS method has been developed for simultaneous measurement of the concentrations of 22 volatile and non-volatile biogenic amines (methylamine, dimethylamine, ethylamine, diethylamine, propylamine, isopropylamine, butylamine, isobutylamine, amylamine, isoamymine, 2-methylbutylamine, hexylamine, pyrrolidine, piperidine, morpholine, 1,3-diaminopropane, putrescine, cadaverine, 1,6-diaminohexane, β-phenylethylamine, tyramine, and histamine) in alcoholic beverages. The method was based on a previously reported two-phase derivatization procedure with isobutyl chloroformate, which reacted quantitatively with all the amines in 10 min. After eliminating excess reagent, by treatment with alkaline methanol or by evaporation (tyramine and histamine determination), the derivatized extracts were analyzed by GC-MS with single-ion monitoring (SIM). Quantitation of amines was achieved by use of a set of eight different internal standards, including five deuterated isotopic analogs. The method has excellent analytical characteristics. It has been used to assay the concentrations of these biogenic amines in thirteen samples of commercially available beers. Eleven biogenic amines were found in all the samples; putrescine (maximum 6.9 mg L−1) and tyramine (maximum 1.7 mg L−1) were the most abundant. Histamine was found in all samples at very low levels (maximum 147 μg L−1).

Concentrations of five biogenic amines in Czech beers and factors affecting their formation

Biogenic amines in beer and in solid raw materials were determined by a high-performance liquid chromatographic method as N-benzamides. In a survey of 78 proprietary bottled bottom-fermented lager beers from 32 Czech breweries, mean concentrations were 0.55, 1.21, 6.85, 8.84 and 12.9 mg/l and standard deviations 1.08, 1.46, 5.19, 7.06 and 12.3 mg litre −1 for histamine, tryptamine, tyramine, putrescine and cadaverine, respectively. No significant differences were observed between beer types (pale/dark, original extract of wort 10%/12%) and amine concentrations. Testing three batches for changes in the amine concentration during the brewing process showed that the formation of amines occurred principally during the main fermentation.

Determination of Biogenic Amines in Beers and Their Raw Materials by Ion-Pair Liquid Chromatography with Postcolumn Derivatization

Abstract A liquid chromatographic method is described for the determination in one run of the following 10 biogenic amines in beers: histamine, tyramine, serotonin, (β-phenylethylamine, tryptamine, cadaverine, putrescine, agmatine, spermine, and spermidine. The method is based on ion-pair chromatographic partition on a reversed-phase column and involves a postcolumn reaction with o-phthalaldehyde (OPT) to form fluorescent derivatives with amines. Treatment of beer samples before injection requires only a filtration through a 0.45 μm filter. It is necessary to obtain a perchloric extract for analysis of raw materials (malt and hops). The method was tested for lack of interference from amino acids and other amines. Linearity, precision, recovery, and sensitivity were satisfactory. Detection limits ranged from 0.30 to 0.40 mg/L, and determination limits ranged from 0.40 to 0.50 mg/L, except for serotonin and spermine, which were slightly higher. The simple preparation of sample and the automatic accomplishment of derivatization steps considerably reduce analysis time and effort.