The analysis of biogenic amines in fresh and processed food is of great importance not only for the potential risk these compounds have on human health, but also because these amines can perform as chemical indicators of food quality and enable the assessment of food processing conditions and/or microbial contamination. A good option for a rapid detection of biogenic amines is the application of biosensors, as these devices enable the obtainment of results in a few minutes without pretreatment of the analyzed material. Biosensors for biogenic amines comprise various combinations of different enzymes for selective biorecognition and signal transduction systems and are based on different signal mechanisms. The present review gives a condensed overview of the recent developments and issues in the construction of biosensors for the detection of most common biogenic amines found in food and other protein-containing material.
Polyphenolic compounds, plant secondary metabolites essential for plant survival, are known for their high antioxidant and anti-inflammatory activity. In addition, several polyphenols, such as phloretin, also have potential antiviral effects, making these compounds potential ingredients of biofunctional foods. A promising source for the extraction of phloretin is a by-product of apple production—apple tree leaves. Focusing on green technologies, the first aim of the present study was to optimize the direct ultrasound-assisted extraction conditions to gain the maximum yield of phloretin from air-dried apple leaves. For the optimization of process parameters, we applied the response surface method with Box–Behnken design. The optimal extraction conditions were extraction time 14.4 min, sonication amplitude 10% and 10 g of sample per 100 mL solvent (70% ethanol, w/w). Using these conditions, we assessed the content of individual and total polyphenolic compounds along with antioxidant activity in the leaves of different autumn and winter apple cultivars grown in Estonia. The analyses were carried out with chromatographic (HPLC-DAD-MS/MS) and spectrophotometric methods. The phloretin concentration ranged from 292 to 726 µg/g and antioxidant activity from 6.06 to 11.42 mg GA eq./g, these being the highest in the local winter cultivars ‘Paide taliõun’ and ‘Tellissaare’, respectively.
Research on extracellular vesicles (EVs) has intensified over the past decade, including fluorescent membrane labeling of EVs. An optimal fluorescent method requires the size of EVs to be preserved after labeling. Lipophilic fluorescent dyes, such as CellMask™ Green (CMG), have been widely used for this purpose. Here, we investigated conditions affecting the optimum CMG labeling of EVs derived from human choriocarcinoma cells (JAr) and different biological fluids using fluorescence NTA (fl-NTA). The effect of CMG labeling on the size, concentration and zeta potential (ZP) on JAr EVs purified with different methods were measured along with biological fluid-derived EVs. With the increase of CMG dye concentration, a significant decrease in the mean size of fluorescent nanoparticles (fl-NPs) was observed. The ZP of fl-NPs originating from JAr cells with the lowest and highest dye concentrations showed a significant shift towards more and less negative ZP values, respectively. Differences in the concentration of fl-NPs were observed for JAr EVs purified using size-exclusion chromatography (SEC) alone and SEC in combination with tangential flow filtration. The proportion of CMG labeling of NPs varied across different biological sources. CMG labeling may be a reliable technique for the detection of EVs using fl-NTA.