Current methods of analysis for the determination of trichothecene mycotoxins in food

This article describes the trends in analytical techniques for the determination of trichothecene mycotoxins, namely deoxynivalenol, and T-2 and HT-2 toxins in cereals and cereal products with particular emphasis on screening and rapid approaches. The driving force behind the changing methodologies is mainly attributed to legislative demands. However, for commercial and governmental testing laboratories, the need to use validated official methods is ever increasing to ensure quality assurance of results.Much research has been undertaken to improve screening assays, highlighted by the number of new methods using a variety of formats and platforms, including optical and electrochemical biosensors. Significant advances in the traditional reference methods have also been demonstrated in addition to the emergence of a variety of commercial immunoaffinity and solid-phase extraction columns for clean up. The use of liquid chromatography coupled to tandem mass spectrometry for mycotoxin detection is ever increasing, allowing simultaneous determination of many toxins in various sample matrices.     

基于纳米抗体-碱性磷酸酶融合蛋白的一步酶联免疫吸附分析法检测黄曲霉毒素B1

从噬菌体展示的抗黄曲霉毒素B1(AFB1)纳米抗体文库中,通过四轮亲和淘选,得到抗AFB1纳米抗体G8.将编码纳米抗体G8的基因与碱性磷酸酶(AP)基因融合,构建了重组融合表达载体pET25b(+)-G8-AP,将其转化大肠杆菌BL21(Rosetta,DE3)感受态细胞,异丙基-β-D-硫代吡喃半乳糖苷(IPTG)诱导融合基因表达.SDS-PAGE结果表明,融合蛋白G8-AP为可溶性表达,Ni2+-NTA亲和层析柱纯化融合蛋白,对硝基苯磷酸二钠(pNPP)法测得纯化后的G8-AP的碱性磷酸酶比活力为(364.5±8.3)U/mg.ELISA检测结果表明,G8-AP具有特异识别AFB1的活性,与其它真菌毒素(AFB2、AFG1、AFG2、脱氧雪腐镰刀菌烯醇、玉米赤霉烯酮、伏马菌素B1)无交叉反应.基于G8-AP建立了检测AFB1的一步ELISA法.在优化的条件下,即甲醇浓度20％、盐离子浓度20 mmol/L、pH 7.4条件下,方法的半数抑制浓度(IC50)为19.8 ng/mL,线性范围为4.3 ～ 92 ng/mL,检出限为2.6 ng/mL.对玉米和小麦样品加标回收实验结果表明,基质对本方法的干扰不明显,可用于实际样品检测.     

生物活性玻璃简介

生物活性玻璃因具有良好的生物相容性、生物活性、骨传导性和可降解性,在化学材料和医学领域都受到广泛的关注。目前,生物活性玻璃已经被成功应用于骨损伤及牙科疾病的治疗和修复等领域。就生物活性玻璃的组成与结构、制备方法、生物活性机理及其在骨修复和牙科领域的应用进行了阐述,并就其发展前景进行了展望。     

基于高特异性单克隆抗体的间接竞争ELISA检测食品中胭脂红的研究

该研究设计合成了一种胭脂红半抗原,分别采用重氮化法和戊二醛法将半抗原与载体蛋白偶联制备人工抗原,通过免疫Bal b/c小鼠及杂交瘤技术成功筛选制备了胭脂红高特异性单克隆抗体,与苋菜红、柠檬黄等结构类似物无交叉反应.基于该抗体建立了间接竞争酶联免疫分析方法用于检测食品中胭脂红残留.该方法对胭脂红的半抑制浓度(IC50)和...     

Verification of the Conditions for Determination of Antioxidant Activity by ABTS and DPPH Assays—A Practical Approach

The ABTS and DPPH methods are among the most popular assays of antioxidant activity determination. Attempts to adapt them to different analytes and the search for the highest values of antioxidant activity has resulted in a large variety of assay conditions to be presented in the literature, including the way the measurement is made. This makes it difficult to relate the results to real oxidation systems, and often makes it impossible to compare them. Such a comparison is limited in advance by the use of stable radicals that do not exist in nature and that react differently from those generated in food or in vivo. Therefore, it is important to introduce measures aimed at standardizing the conditions of the activity assay, including reaction time and several reaction environments suitable for testing different groups of compounds. In this study, we used natural antioxidants of various structures: phenolic acids, flavonoids, peptides and corresponding amino acids, ascorbic acid and α-tocopherol, and also synthetic analogues of selected compounds. The curves of dependence of the measured absorbance on the concentration of antioxidants were described, the ranges of linearity were determined, and the value of the error made when reading in various ranges of dependencies was estimated. We also determined and compared the activity values using two popular methods (IC₅₀ and TEAC), taking into account different environments and reaction times. Based on the collected data, recommendations were formulated regarding the reaction conditions adapted to the studies of individual groups of antioxidants, and unified reaction times were proposed. Taking into account the state before reaching the equilibrium of antioxidants reacting in a complex manner, this approach may introduce a simplified reference to the competing reaction that occurs in reality.     

Anti-α-Glucosidase and Antiglycation Activities of α-Mangostin and New Xanthenone Derivatives: Enzymatic Kinetics and Mechanistic Insights through In Vitro Studies

Diabetes mellitus is characterized by chronic hyperglycemia that promotes ROS formation, causing severe oxidative stress. Furthermore, prolonged hyperglycemia leads to glycation reactions with formation of AGEs that contribute to a chronic inflammatory state. This research aims to evaluate the inhibitory activity of α-mangostin and four synthetic xanthenone derivatives against glycation and oxidative processes and on α-glucosidase, an intestinal hydrolase that catalyzes the cleavage of oligosaccharides into glucose molecules, promoting the postprandial glycemic peak. Antiglycation activity was evaluated using the BSA assay, while antioxidant capacity was detected with the ORAC assay. The inhibition of α-glucosidase activity was studied with multispectroscopic methods along with inhibitory kinetic analysis. α-Mangostin and synthetic compounds at 25 µM reduced the production of AGEs, whereas the α-glucosidase activity was inhibited only by the natural compound. α-Mangostin decreased enzymatic activity in a concentration-dependent manner in the micromolar range by a reversible mixed-type antagonism. Circular dichroism revealed a rearrangement of the secondary structure of α-glucosidase with an increase in the contents of α-helix and random coils and a decrease in β-sheet and β-turn components. The data highlighted the anti-α-glucosidase activity of α-mangostin together with its protective effects on protein glycation and oxidation damage.     

磁性侧流层析技术在食品污染物快速检测中的研究进展

磁性侧流层析技术（MLFA）是一种将磁性纳米材料与侧流层析检测平台相结合的快速分析技术。在保留了传统侧流层析技术操作简单、快速实时、低成本优势的基础上,引入功能化磁性纳米材料,实现了复杂基质中目标物的选择性分离和富集,从而提高检测灵敏度和准确性。该文重点关注了近五年来MLFA技术在食品分析领域的研究进展,总结了磁性纳米材料的主要种类及MLFA的主要检测原理,并对食源性病原菌和真菌毒素、兽药等小分子有害物质的检测进行了介绍。最后,总结了MLFA技术的研究现状并对未来的发展方向进行了展望。     

Design,Synthesis and Bioactivity Evaluation of Coumarin–BMT Hybrids as New Acetylcholinesterase Inhibitors

Coumarin possesses the aromatic group and showed plentiful activities, such as antioxidant, preventing asthma and antisepsis. In addition, coumarin derivatives usually possess good solubility, low cytotoxicity and excellent cell permeability. In our study, we synthesized the compound bridge methylene tacrine (BMT), which has the classical pharmacophore structure of Tacrine (THA). Based on the principle of active substructure splicing, BMT was used as a lead compound and synthesized coumarin–BMT hybrids by introducing coumarin to BMT. In this work, 21 novel hybrids of BMT and coumarin were synthesized and evaluated for their inhibitory activity on AChE. All obtained compounds present preferable inhibition. Compound 8b was the most active compound, with the value of K_i as 49.2 nM, which was higher than Galantamine (GAL) and lower than THA. The result of molecular docking showed that the highest binding free energy was −40.43 kcal/mol for compound 8b, which was an identical trend with the calculated K_i.     

Oxidative Stress and DNA Damage Effect of Dioscorea hispida Dennst. on Placental Tissues of Rats

Dioscorea hispida Dennst. locally known as “ubi gadung” has been used as a traditional remedy and source of carbohydrate among Malaysians. To assess the effect of Dioscorea hispida aqueous extract (DHAE) on the production of reactive oxygen species (ROS) and their effects on DNA damage in Sprague Dawley rat’s placental tissues, pregnant rats were randomly divided into four groups. The animals were orally treated with distilled water (negative control) and three different concentrations of DHAE (250, 500 and 1000 mg/kg body weight (BW)) from gestation day 6 until 20. The oxidative stress in placental tissues was evaluated at day 21 by measuring the level of ROS, superoxide dismutase (SOD) and lipid peroxidation biomarker, malondialdehyde (MDA) while comet assay was used for DNA damage. There was no significant production of ROS and SOD activities in all groups. Significant changes were observed in the MDA level at 1000 mg/kg BW DHAE. Comet assay revealed a significant increase (p < 0.05) of DNA damage on animals treated with 250 and 500 mg/kg BW DHAE but not at the highest concentration. It was postulated that the placental cells could have undergone necrosis which destroys all components including DNA. This occurrence simultaneously reduces the levels of DNA damage which can be represented by lower level of tail moments. This finding correlates with our histopathological examination where necrotic cells of spongiotrophoblast were observed in the basal zone of placental tissue. The high amount of hydrogen cyanide and other compounds in 1000 mg/kg BW DHAE could elevate the lipid peroxidation and directly induce cell necrosis which requires further investigation.     

Novel Chemical and Biological Insights of Inositol Derivatives in Mediterranean Plants

Inositols (Ins) are natural compounds largely widespread in plants and animals. Bio-sinthetically they derive from sugars, possessing a molecular structure very similar to the simple sugars, and this aspect concurs to define them as primary metabolites, even though it is much more correct to place them at the boundary between primary and secondary metabolites. This dichotomy is well represented by the fact that as primary metabolites they are essential cellular components in the form of phospholipid derivatives, while as secondary metabolites they are involved in a plethora of signaling pathways playing an important role in the surviving of living organisms. myo-Inositol is the most important and widespread compound of this family, it derives directly from d-glucose, and all known inositols, including stereoisomers and derivatives, are the results of metabolic processes on this unique molecule. In this review, we report the new insights of these compounds and their derivatives concerning their occurrence in Nature with a particular emphasis on the plant of the Mediterranean area, as well as the new developments about their biological effectiveness.