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.     

Biogenic amines profile and concentration in commercial milks for infants and young children

Commercial milks for infants and young children (CMIYC) received much attention during last years for their impact on the nutritional status, health and development of the new-born and babies. Among possible contaminants contained in these foods, biogenic amines (BAs) have rarely been determined although they can exert toxic effects in humans if ingested at high concentrations. Spermine, spermidine, putrescine, histamine, tyramine, β-phenylethylamine and cadaverine have been quantified in CMIYC samples by LC-UV after derivatisation with dansyl-chloride. Once optimised in terms of linearity (R² ≥ 0.989), recovery percentages (92.9–97.3), LOD (0.2–0.4 μg g^?1 or 0.03–0.05 μg mL^?1 depending on the samples), LOQ (0.5–1.0 μg g^?1 and 0.08–0.13 μg mL^?1 depending on the samples) and repeatability (0.1–0.2 intra-day; 0.2–0.4 inter-day), the method has been applied to real samples. Very low total BAs concentrations have been found in reconstituted (1.18–3.12 mg L^?1) and liquid milks (0.33–2.30 mg L^?1), with different biogenic amine profiles and distributions. A risk assessment based on the available information regarding Acute Reference Doses of histamine and tyramine, as well as the application of common Biogenic Amine Indexes, showed that none of the analysed samples represented a possible risk for babies, also considering a worst case evaluation. These findings confirmed the strict safety and quality protocols adopted during the production of CMIYC.

Chemical compounds studied in this article: Ammonium chloride (PubChem CID: 25517); Cadaverine hydrochloride (PubChem CID: 5351467); Hydrochloridric acid (PubChem CID: 313); Histamine dihydrochloride (PubChem CID: 5818); Phenylethylamine hydrochloride (PubChem CID: 9075); Putrescine dihydrochloride (PubChem CID: 9532); Sodium hydroxide (PubChem CID: 14798); Spermine tetrahydrochloride (PubChem CID: 1103); Spermidine trihydrochloride (PubChem CID: 1102); Tyramine hydrochloride (PubChem CID: 66449).     

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).     

Determination of biogenic amines in traditional Chinese fermented foods by reversed-phase high-performance liquid chromatography (RP-HPLC)

A survey of biogenic amine content of traditional Chinese fermented foods (douchi, sufu, fermented sausage, yulu, and shrimp paste) was carried out. Eight major biogenic amines including putrescine, cadaverine, histamine, phenylethylamine, tyramine, spermine, spermidine and tryptamine were separated by reversed-phase HPLC-DAD on Inertsil ODS-SP column after pre-column derivatisation with dansyl chloride. The results showed that spermine (1.65–3.96 mg 100 g^?1), putrescine (0.20–10.89 mg 100 g^?1), cadaverine (3.60–12.14 mg 100 g^?1), and histamine (0.57–20.24 mg 100 g^?1) were the most represented amines. All amines were detected in yulu, while sufu showed a much higher content of most amines. Shrimp paste showed the lowest level of total biogenic amines. Moreover, the composition and content of eight biogenic amines in the selected samples varied among different food types, origins and batches. Although the average content for each amine is within the range that may elicit direct adverse reactions, consumers should be aware of the potential synergistic effect among different amines and limit their consumption at each meal.     

Effects of gamma irradiation on the biogenic amines in pepperoni with different packaging conditions

Combined effects of gamma irradiation and packaging on biogenic amine (BA) formation in pepperoni sausage were investigated during storage. Pepperoni (fermented sausage) was made and packaged with air, vacuum and CO₂/N₂ (25%/75%) gas and then gamma-irradiated at 0, 5, 10 and 20 kGy. The pH was decreased after storage and non-irradiated samples showed a lower pH than irradiated. Lactic acid bacteria (LAB) were not detected after 5 kGy of gamma irradiation, while the LAB in the non-irradiated samples were 4–5 log counts during storage. A total of six different BAs, putrescine, cadaverine, β-phenylethylamine, spermidine, spermine and tyramine, were found in the pepperoni sausage. Detected BAs were statistically low under the irradiation or packaging conditions, except for cadaverine and β-phenylethylamine. Gamma irradiation was effective in reducing putrescine, spermidine, spermine and tyramine. Irradiation effects were not observed on the β-phenylethylamine level, while the CO₂/N₂ packaging caused increasing of the level. Most BAs detected were reduced by gamma irradiation of the pepperoni sausage during storage. However, CO₂/N₂ packaging was an improper condition for BA reduction in the pepperoni 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.     

Aluminium,nickel, cadmium and lead in candy products and assessment of daily intake by children in Spain

Aluminium (Al), nickel (Ni), cadmium (Cd) and lead (Pb) levels in a total of 263 samples of 12 types of candies widely consumed in Spain, were evaluated. Samples were analysed using an ICP-MS method after acidic sample mineralization. Concentration ranges of Al, Ni, Cd and Pb were 21.28–62.91 µg g^?1, 0.40–1.27 µg g^?1, 0.12–1.01 µg g^?1 and 1.03–7.14 µg g^?1, respectifgvely. Statistically significant positive correlations were calculated between concentrations of Ni–Al and Pb–Cd (p-values < 0.05). Taking into consideration the relatively high metal content, together with the high caloric density of these products, as well as high content of particular nutrients such as sugars that can induce development of certain pathologies like obesity and caries, indicate that frequent consumption of candy products is not recommended.     

Biogenic amines in alcoholic beverages produced in Taiwan

The biogenic amines tyramine, putrescine, cadaverine, 2-phenylethylamine, histamine and tryptamine were determined in 13 kinds of alcoholic beverages produced in Taiwan. They were derivatised with dansyl chloride and analysed using a high-performance liquid chromatographic method. Total amine content ranged from 0.23 to 11.4 μg ml^?1 with Charng Chuen liqueur containing the highest level and Mei Kwei Lu the lowest. Tryptamine was found in all samples in amounts between 0.01 and 1.79 μg ml^?1. The levels of the other five amines in the tested samples were 0-7.0 μg ml^?1 for 2-phenylethylamine, 0-3.0 μg ml^?1 for putrescine, 0-2.9 μg ml^?1 for cadaverine, 0-4.5 μg ml^?1 for histamine, and 0-1.4 μg ml^?1 for tyramine. These levels seem unlikely to have adverse effects on human health.     

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.     

Investigation of pyrrolizidine alkaloids including their respective N-oxides in selected food products available in Hong Kong by liquid chromatography electrospray ionisation mass spectrometry

ABSTRACT

This study determined the levels of pyrrolizidine alkaloids (PAs), including their respective N-oxides, in foodstuffs available in Hong Kong by liquid chromatography-electrospray ionisation tandem mass spectrometry. A total of 234 samples (48 food items) were collected randomly from a local market and analysed. About 50% of samples were found to contain detectable amount of PAs. Amongst the 48 food items, PAs were not detected in 11 food items, including barley flour, beef, cattle liver, pork, pig liver, chicken meat, chicken liver, milk, non-fermented tea, Melissa tea and linden tea. For those found to contain detectable PAs, the summed PA content ranged up to 11,000 µg kg^?1. The highest sum of PA content among the 37 food items calculated with lower bound was cumin seed, then followed by oregano, tarragon and herbs de Provence with ranges of 2.5–11,000, 1.5–5100, 8.0–3300 and 18–1300 µg kg^?1 respectively. Among the samples, the highest sum of PA content was detected in a cumin seed sample (11,000 µg kg^?1), followed by an oregano (5100 µg kg^?1), a tarragon (3300 µg kg^?1) and a herbs de Provence (1300 µg kg^?1). In general, the results of this study agreed well with other published results in peer-reviewed journals, except that the total PAs in honey and specific tea infusion in this study were comparatively lower.     

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.     

Occurrence of ochratoxin A and biogenic amines in Asian beers sold in French markets

Fifty‐one samples of 17 Asian beer brands commercially available in France were analysed for the occurrence of ochratoxin A and biogenic amines, including ethanolamine, histamine, methylamine, ethylamine, tyramine, tryptamine, 2‐phenylethylamine, putrescine, isopenthylamine and cadaverine. The types and levels of biogenic amines varied from one brand to another. None of the ochratoxin A or biogenic amine contaminants in the Asian beer samples exceeded the recommended levels for safe consumption. Ochratoxin A was found to range from n.d. to 0.175 µg L^?1. Biogenic amines were detected in all samples at a level ranging from 6.15 to 47.30 mg L^?1. Isoamylamine was not detected in any of the beers. Ethanolamine, tyramine, putrescine, and cadaverine were the most abundant of the nine biogenic amines found in this study. The concentrations were 2.75–12.88 mg L^?1 for ethanolamine, 0.58–17.37 mg L^?1 for tyramine, 1.54‐9.78 mg L^?1 for putrescine and 0.31–11.39 mg L^?1 for cadaverine. The work revealed significant, strong correlations (p < 0.01) between concentrations of ochratoxin A and biogenic amines, especially tyramine, tryptamine and cadaverine. Copyright © 2013 The Institute of Brewing & Distilling     

Biogenic amines in commercially produced Yulu,a Chinese fermented fish sauce

Seven biogenic amines were determined in 35 commercially produced Yulu samples from three provinces of China by pre-column derivatisation with dansyl chloride (Dns-Cl) and high-performance liquid chromatography with fluorescence detection (HPLC-FLD). Putrescine, cadaverine, histamine and tyramine were the major biogenic amines (more than 100 mg kg^?1), while tryptamine, spermidine and spermine were regarded as minor biogenic amines (less than 25 mg kg^?1). Twenty samples contained more than 50 mg kg^?1 histamine (the limit for histamine in seafood products as suggested by the Food and Drug Administration). Twenty-one samples contained more than 100 mg kg^?1 tyramine and 10 contained more than 1000 mg kg^?1 total biogenic amines. This study provided data on biogenic amine levels in Chinese fermented fish sauce. The results suggested that biogenic amine content should be monitored in commercially produced Yulu.     

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).     

Determination and surveillance of hydrocortisone and progesterone in livestock products by liquid chromatography-tandem mass spectrometry

A simple analytical method for the determination of hydrocortisone and progesterone in bovine, swine, and chicken muscle and eggs was developed. Hydrocortisone and progesterone were extracted with acetonitrile and subsequently cleaned-up using an Oasis^® HLB mini-cartridge. The method was validated in accordance with Japanese guidelines and exhibited trueness from 86.6% to 104.3% and precision (relative standard deviations (RSDs) of repeatability and within reproducibility were under 8.7% and 11.7%, respectively). The method was applied to 103 bovine muscle, 137 swine muscle, 69 chicken muscle and 52 egg samples that were commercially available in Tokyo, Japan. The hydrocortisone concentration was 0.9–41.2 µg kg^?1 in all bovine muscle samples, with an average of 7.7 µg kg^?1 and a median of 6.2 µg kg^?1. The progesterone concentration in 50 samples exceeded the limit of quantification (LOQ) and reached a maximum of 95.4 µg kg^?1. Hydrocortisone was also detected in all swine muscle samples at concentrations of 2.0–56.0 µg kg^?1. Its average and median concentrations amounted to 13.1 and 11.3 µg kg^?1, respectively. Twenty-three samples contained progesterone levels surpassing the LOQ, with a maximum concentration of 107.0 µg kg^?1. No chicken muscle samples contained any of the analytes. The progesterone concentration was 15.5–200.0 µg kg^?1 in all egg samples, with an average of 95.4 µg kg^?1 and a median of 90.5 µg kg^?1.     

Aflatoxins and ochratoxin A in export quality raisins collected from different areas of Pakistan

During 2012–2014, 170 samples of export quality raisins were collected from different vendors in Pakistan. The collected samples were analysed for the presence of aflatoxins (AFs) and Ochratoxin A (OTA) contamination using high-performance liquid chromatography technique. The limit of detection and limit of quantification of AFs/OTA were 0.12/0.10 and 0.36/0.30 µg kg^?1, respectively. Only 5% of the samples were contaminated with AFs, ranging 0.15–2.58 µg kg^?1 with a mean of 0.05 ± 0.26 µg kg^?1. None of the raisin samples exhibited AFs contamination above the maximum limit (ML = 4 µg kg^?1) as set by the European Union (EU). About 72% of the samples were contaminated with OTA, ranging 0.14–12.75 µg kg^?1 with a mean of 2.10 ± 1.9 µg kg^?1. However, in 95.3% of the tested samples, OTA level was lower than the ML of 10 µg kg^?1 as regulated by the EU. Apparently, a strict and continuous monitoring plan, including regulatory limits, improves food safety and quality for all types of commodities.     

Benzoic and sorbic acid in soft drink,milk, ketchup sauce and bread by dispersive liquid–liquid microextraction coupled with HPLC

Benzoic acid and sorbic acid are widely used for food preservation. These preservatives are generally recognised as safe. The aim of this study was to determine the level of benzoic and sorbic acid in food samples that are usually consumed in Iran. Therefore, 54 samples, including 15 soft drinks, 15 ultra-high-temperature milk, 15 ketchup sauces and 9 bread samples, were analysed by high-performance liquid chromatography with UV detection. Benzoic acid was detected in 50 (92.5%) of the samples ranging from 3.5 to 1520 µg mL^?1, while sorbic acid was detected in 29 (50.3%) samples in a range of 0.8 and 2305 µg mL^?1. Limits of detection and limits of quantification for benzoate were found to be 0.1 and 0.5 µg mL^?1, respectively, and for sorbate 0.08 and 0.3 µg mL^?1, respectively. The results showed that benzoic acid and sorbic acid widely occur in food products in Iran.     

Tropane and ergot alkaloids in grain-based products for infants and young children in the Netherlands in 2011–2014

An LC-MS/MS multi-method was developed to simultaneously quantify ergot alkaloids (EAs) and tropane alkaloids (TAs) in 113 cereal-based food for infants and young children. To assess yearly variation, samples were collected in 2011, 2012 and 2014. EAs were detected in 54% and TAs in 22% of the samples. Mean EA levels in the three sampling years were 10.6, 6.2 and 8.6 µg kg^?1, respectively (maximum: 115.4 µg kg^?1), indicating that exposure to EAs would not have exceeded the health-based guidance values set by EFSA in 2012. Mean TA levels were 3.9, 2.4 and 0.4 µg kg^?1, respectively (maximum: 80.8 µg kg^?1). The acute reference dose for TAs, derived by EFSA in 2013, would have been exceeded by young children when consuming some of the products sampled in 2011–2012. TA levels had decreased drastically in 2014, possibly due to measures taken by producers as response to the EFSA Opinion.     

Variation in content of tyramine,histamine, 2-phenylethylamine,tryptamine and their precursor amino acids in a Chardonnay wine by using different commercial active dry yeasts and nitrogen sources

Wine is a fermented beverage that could be affected by high concentrations of biogenic amines, thus altering organoleptic and health properties. In this work, the capacity of different selected yeast strains regarding to affect the content of tyramine, histamine, 2-phenylethylamine, tryptamine and their precursor amino acids (pAA) during fermentation has been described. Overall, biogenic amines (BAs) and pAA contents appeared significantly related both to the strain and to the nutrient supplementation applied. Among BAs, without and (with) nitrogen source addition, the concentrations covered a range between 0.1 and 9.5 (0.1–10.3) mg L⁻¹ for tyramine; 0.1 and 4.4 (0.1–4.6) mg L⁻¹ for histamine; 0.4 and 1.1 (0.4–1.4) mg L⁻¹ for 2-phenylethylamine; and 0.02 and 0.14 (0.01–0.12) mg L⁻¹ for tryptamine, respectively. Considering that also wine yeast species are responsible for BAs formation, the evidence from fermentation trials underlined the relevance of a careful choice of ADY strains and nitrogen sources in the management of alcoholic fermentation in wine.     

Deoxynivalenol in wheat,maize, wheat flour and pasta: surveys in Hungary in 2008–2015

Among Fusarium mycotoxins, deoxynivalenol (DON) is the most common contaminant in case of cereals and cereal-based foods in Hungary. In this study, Hungarian wheat (n = 305), maize (n = 108), wheat flour (n = 179) and pasta (n = 226) samples were analysed (N = 818). The samples were collected during 2008–2015 in Hungary. Applied methods of analysis were enzyme-linked immunosorbent assay and liquid-chromatography coupled with a mass spectrometer. Results were compared and evaluated with Hungarian weather data. Among cereal samples, in 2011, wheat was contaminated with DON (overall average ± standard deviation was 2159 ± 2818 µg kg^?1), which was above the maximum limit (ML). In case of wheat flour and pasta, no average values above ML were found during 2008–2015, but higher DON contamination could be observed in 2011 as well (wheat flour: 537 ± 573 µg kg^?1; pasta: 511 ± 175 µg kg^?1).