| Chromatographic Analysis for Targeted Metabolomics of Antioxidant and Flavor-Related Metabolites in Tomato
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Author:
Date:
2021-03-05
[Abstract] Targeted metabolomics is a useful approach to evaluate crop breeding studies. Antioxidant and flavor-related traits are of increasing interest and are considered quality traits in tomato breeding. The present study presents chromatographic methods to study antioxidants (carotenoids, vitamin C, vitamin E, phenolic compounds, and glutathione) and flavor-related characters (sugars and organic acids) in tomato. Two different extraction methods (for polar and apolar entities) were applied to isolate the targeted compounds. The extraction methods developed in this work were time and cost-effective since no further purification was needed. Carotenoids, vitamin C, glutathione, and phenolic acids were analyzed by HPLC-PDA using a RP C18 column at an appropriate wavelength for each compound. ...
[摘要] [摘要]靶向代谢麦克风是评估作物育种研究的有用方法。抗氧化剂和与风味有关的性状受到越来越多的关注，被认为是番茄育种中的品质性状。本研究提供了色谱方法来研究番茄中的抗氧化剂（类胡萝卜素，维生素C，维生素E，酚类化合物和谷胱甘肽）和与风味相关的特征（糖和有机酸）。应用了两种不同的提取方法（针对极性和非极性实体）来分离目标化合物。由于不需要进一步纯化，因此这项工作中开发的提取方法既省时又经济。使用RP C18色谱柱，通过RP-C18色谱柱在适当波长下对每种化合物进行类胡萝卜素，维生素C，谷胱甘肽和酚酸的分析。用RP C18和NH2色谱柱通过HPLC分析维生素E和糖，并分别通过FLD和RI检测器进行检测。此外，MSTFA衍生化后，使用Rtx 5DA色谱柱使用GC-FID分析有机酸。结果，开发了定量重要植物代谢物的灵敏分析方法，并在本文中进行了描述。这些方法不仅适用于番茄，但都还表征其他物种的香味相关的和抗氧化的化合物。因此，这些协议可用于指导作物育种的选择。
[背景]代谢组学是其受到关注的其潜在的援助作物育种研究施加生化途径。在番茄育种中，由于消费者对提高作物品质的偏好，抗氧化剂和与风味有关的代谢产物都受到了越来越多的关注。人类无法自己合成抗氧化剂分子; ...
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| RETRACTED: Paper Lateral Flow Biosensor for Nodavirus Reverse Transcribed RNA Detection
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Author:
Date:
2020-08-05
[Abstract] Paper nanobiosensors have been established as an excellent platform for analysis of veterinary and human pathogens causing various diseases. Especially, lateral flow assays or biosensors ideal for sensitive, rapid, robust and accurate analysis in laboratory setups and on-site analysis. Viral RNA detection is of great importance for public health as well as animal health protection. In that aspect, the present protocol focuses on the development of functionalized gold nanoparticle-based lateral flow biosensor for fish nervous necrosis virus (Nodavirus) nucleic acids detection. Total viral RNA, isolated from fish samples was subjected to reverse transcription PCR amplification and the amplification products were mixed with specific oligonucleotide probe. A red test line was formed when ...
[摘要] [摘要] 纸纳米生物传感器已经成为分析导致各种疾病的兽医和人类病原体的一个极好的平台。尤其是侧向流分析或生物传感器是实验室设备和现场分析中灵敏、快速、可靠和准确分析的理想选择。病毒RNA检测对公共卫生和动物健康保护具有重要意义。在这一方面，本协议的重点是开发功能化金纳米粒子侧流生物传感器，用于鱼类神经坏死病毒（Nodavirus）核酸检测。从鱼类标本中分离出的总病毒RNA进行逆转录PCR扩增，扩增产物与特异性寡核苷酸探针混合。当有nodavirus产物存在时，形成红色检测线。这种方法对基础研究有很大的意义，因为它消除了耗时、繁琐的电泳程序的需要，并且可以在养鱼场对养鱼户进行调整。利用这种生物分析平台进行病害监测，无需耗费大量时间和成本，对水产养殖和环境安全有很大影响。
[背景] 关注点和/或现场生物分析一直是关注人类和动物福祉的研究工作的最终目标。基于纸基的传感平台具有功能化简单、重现性好、制造成本低等优点，是一种极具吸引力的分析平台。纸基分析设备已应用于小分子、蛋白质和各种核酸的分析（Parolo和Merkoçi，2013；Bahadir和Sezgintürk，2016；Jiang等人，2019）。侧流生物传感器（LFB）是一种带有干试剂的预制材料条带，通过流体样品激活。它们专为一次性一次性使用而设计，只要有足够的开/关信号（Posthuma ...
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| Bacterial Cell Wall Precursor Phosphatase Assays Using Thin-layer Chromatography (TLC) and High Pressure Liquid Chromatography (HPLC)
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Author:
Date:
2018-03-20
[Abstract] Peptidoglycan encases the bacterial cytoplasmic membrane to protect the cell from lysis due to the turgor. The final steps of peptidoglycan synthesis require a membrane-anchored substrate called lipid II, in which the peptidoglycan subunit is linked to the carrier lipid undecaprenol via a pyrophosphate moiety. Lipid II is the target of glycopeptide antibiotics and several antimicrobial peptides, and is degraded by ‘attacking’ enzymes involved in bacterial competition to induce lysis. Here we describe two protocols using thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC), respectively, to assay the digestion of lipid II by phosphatases such as Colicin M or the LXG toxin protein TelC from Streptococcus intermedius. The TLC method can also monitor the ...
[摘要] 肽聚糖包裹细菌细胞质膜以保护细胞免于因膨胀而导致的溶解。 肽聚糖合成的最后步骤需要称为脂质II的膜锚定底物，其中肽聚糖亚基通过焦磷酸部分连接至载体脂质十一碳烯醇。 脂质II是糖肽抗生素和几种抗微生物肽的靶标，并且通过参与细菌竞争的“攻击”酶来降解以诱导裂解。 在这里，我们分别描述了两种使用薄层色谱法（TLC）和高压液相色谱法（HPLC）的方案来测定磷脂酶如Colicin M或来自中间链球菌的LXG毒素蛋白TelC对脂质II的消化，的。 TLC方法也可以监测十一异戊二烯基（pyro）磷酸盐的消化，而HPLC方法允许分离脂质II的二 - ，单 - 或非磷酸化二糖五肽产物。
【背景】肽聚糖（PG）球囊是一种必需的细菌大分子，它可以保护细胞免受由于其膨胀引起的破裂并保持细胞的形状（Vollmer和Bertsche，2008; Typas等人，2012）。 PG由通过短肽连接的聚糖链组成。来自不同物种的PG在肽的结构和二级修饰的存在方面有所不同（Vollmer等人，2008）。 ...
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