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Colorimetric probe for the determination of osmium through a novel optical sensor in environmental samples 通过新型光学传感器测定环境样品中锇含量的比色探针
Q3 Chemistry Pub Date : 2024-03-17 DOI: 10.1016/j.talo.2024.100311
Reem F. Alshehri , Alaa S. Amin , Eman R. Darwish

A novel optical probe has been successfully developed for the selective detection of Os(VIII) ions. This osmium-sensing platform was intricately designed by incorporating 4-(thiazol-2-yldiazenyl)benzene-1,3-diol (TDBD) as the ionophore within a plasticized PVC membrane, and tributylphosphate (TBP) serving as the plasticizer. Upon exposure to Os(VIII) ions in a 0.5 M HClO4 acid milieu, the sensing membrane undergoes a distinctive chromatic transition, shifting from a yellow to a pink hue. The preparation of the sensor and the methodology for Os(VIII) determination were meticulously refined to achieve optimal performance. Under these carefully optimized experimental conditions, the proposed optode demonstrated an impressive linear detection range spanning from 2.5 × 10−9 to 2.5 × 10−5 M for Os(VIII), alongside remarkable detection and quantification limits of 7.24 × 10−10 and 2.4 × 10−9 M, respectively. The sensor consistently delivered highly reproducible results, as evidenced by relative standard deviation (RSD) values of 1.45% and 1.30% for Os(VIII) concentrations at 7.5 × 10−7 and 2.5 × 10−6 M, respectively, underscoring its precision and reliability. Furthermore, the sensor exhibited a swift response time of 3.0 min, further emphasizing its efficiency. The incorporation of PIMs into the sensor architecture led to a remarkable eight-fold enhancement in its response compared to counterparts lacking PIMs, signifying a significant performance improvement. In assessing potential interference, an investigation into the influence of other ions on Os(III) determination revealed that the prepared sensor displayed exceptional selectivity for Os(VIII) ions, with negligible responses observed in the presence of common ions. Collectively, these experimental findings underscore the sensor's efficacy as an invaluable tool for the precise analysis of osmium content in water samples, aligning it with the rigorous standards of international scientific research.

我们成功开发了一种新型光学探针,用于选择性检测 Os(VIII) 离子。这种锇传感平台设计精巧,将 4-(噻唑-2-基二氮烯)苯-1,3-二醇(TDBD)作为离子载体加入塑化聚氯乙烯膜中,并以磷酸三丁酯(TBP)作为增塑剂。在 0.5 M HClO4 酸环境中接触 Os(VIII) 离子时,传感膜会发生明显的色变,从黄色变为粉红色。为了达到最佳性能,我们对传感器的制备和 Os(VIII)的测定方法进行了细致的改进。在这些精心优化的实验条件下,拟议的光电二极管对 Os(VIII) 的线性检测范围从 2.5 × 10-9 M 到 2.5 × 10-5 M,检测限和定量限分别为 7.24 × 10-10 M 和 2.4 × 10-9 M。该传感器始终能提供可重复性很高的结果,在 Os(VIII) 浓度为 7.5 × 10-7 M 和 2.5 × 10-6 M 时,其相对标准偏差 (RSD) 值分别为 1.45% 和 1.30%,这证明了它的精确性和可靠性。此外,该传感器的快速反应时间为 3.0 分钟,进一步提高了其效率。在传感器结构中加入 PIM 后,其响应速度比未加入 PIM 的同类产品显著提高了八倍,这标志着传感器性能的显著提高。在评估潜在干扰时,对其他离子对 Os(III)测定的影响进行了调查,结果表明制备的传感器对 Os(VIII)离子具有极高的选择性,在普通离子存在的情况下,其响应可以忽略不计。总之,这些实验结果证明,该传感器是精确分析水样中锇含量的宝贵工具,符合国际科学研究的严格标准。
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引用次数: 0
Monosaccharide composition of lignocellulosic matrix—Optimization of microwave-assisted acid hydrolysis condition 木质纤维素基质的单糖成分--微波辅助酸水解条件的优化
Q3 Chemistry Pub Date : 2024-03-16 DOI: 10.1016/j.talo.2024.100312
Francesca Modugno, Jeannette Jacqueline Łucejko

Compositional analysis of polysaccharides is crucial in many applications of analytical chemistry, and typically involves hydrolysis as initial step, to break glycosidic bonds and release individual monosaccharide units. We developed a new method for the microwave-assisted acid hydrolysis of polysaccharides, here applied for determining the composition of the polysaccharide fraction within the lignocellulosic matrix of Pinus sylvestris. The optimisation of reaction conditions was carried out using a GC–MS method after a double-step sugar derivatization process.

Under the optimized conditions, specifically, employing 2 M TFA, 40 min, and 120 °C, a high depolymerisation yield was achieved with minimal degradation. Furthermore, the amount of polysaccharides detected in pine wood aligns with the results in existing literature, substantiating the determined chemical composition.

The synergy of microwave-assisted acid hydrolysis and full factorial design not only facilitated the determination of the optimal hydrolysis conditions but also enabled the assessment of the parameters influencing the polysaccharide hydrolysis process and their relationships. This approach made it feasible to achieve maximum hydrolysis efficiency while minimizing analyte degradation, employing a method that reduces environmental impact through the use of microwave energy. This strategy effectively reduced the required reaction temperature, time, and reagent quantities.

多糖的成分分析在分析化学的许多应用中至关重要,通常需要水解作为初始步骤,以打断糖苷键并释放单个单糖单元。我们开发了一种微波辅助酸水解多糖的新方法,并将其用于测定欧洲赤松木质纤维素基质中的多糖成分。在优化的条件下,特别是采用 2 M TFA、40 分钟和 120 °C,可获得较高的解聚率,且降解程度极低。微波辅助酸水解和全因子设计的协同作用不仅有助于确定最佳水解条件,还能评估影响多糖水解过程的参数及其关系。这种方法既能实现最高的水解效率,又能最大限度地减少分析物的降解,还能通过使用微波能减少对环境的影响。这一策略有效降低了所需的反应温度、时间和试剂量。
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引用次数: 0
Advanced review of the contributing factors for the Microwave Digestion of food matrices for trace elemental analysis 微波消解食品基质进行痕量元素分析的诱因高级审查
Q3 Chemistry Pub Date : 2024-03-14 DOI: 10.1016/j.talo.2024.100309
Aisling Sheehan, Ambrose Furey

Microwaves are electromagnetic waves which move at the speed of light. Samples directly exposed to this phenomenon are heated faster than conventional conduction heating methods. This is exploited in acid assisted digestion where a food sample in a suitable acid matrix can be completely destroyed leaving a clear solution of measurable elements. In pressurised systems the temperature can be rapidly increased far above the conventional boiling point of the solvent and the closed microwave vessel environment is ideal for volatile elements.

There is a constant aspiration to achieve lower levels of detection and advances in instrument capabilities with specialised plasma based techniques for the identification and quantification of individual elements at ultra-trace levels have evolved. To realise this; sample preparation, digestion efficiency and sample introduction need to be optimised. Exogenous inputs such as preparation environment, reagent grade, the analyst and apparatus cleaning steps all have the potential to leave an elemental footprint on the sample and therefore contribute to the uncertainty of the analytical result. Knowledge of sample composition and subsequent interaction with microwaves and plasma are an important consideration for complete digestion without which the result is not accurate.

微波是以光速运动的电磁波。与传统的传导加热方法相比,直接暴露在这种现象下的样品加热速度更快。在酸辅助消化中,食物样品在适当的酸基质中会被完全破坏,留下含有可测量元素的透明溶液。在加压系统中,温度可以迅速升高,远远超过溶剂的传统沸点,封闭的微波容器环境非常适合挥发性元素的检测。为实现这一目标,需要对样品制备、消化效率和样品引入进行优化。外源输入,如制备环境、试剂等级、分析师和仪器清洁步骤,都有可能在样品上留下元素足迹,从而导致分析结果的不确定性。了解样品成分以及随后与微波和等离子体的相互作用是完全消解的重要考虑因素,否则结果就不准确。
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引用次数: 0
Hydrophobic barrier-free laminated paper-based analytical device (LPAD) using a diameter-based measurement for determination of iodide in pharmaceutical products 利用基于直径的测量方法测定药品中碘化物含量的疏水无障碍层压纸基分析装置 (LPAD)
Q3 Chemistry Pub Date : 2024-03-09 DOI: 10.1016/j.talo.2024.100310
Nakarin Noirahaeng , Jirawat Salungyu , Saowapak Teerasong , Kanchana Uraisin , Phoonthawee Saetear

This work presents, for the first time, an exploitation of an argentometric Mohr's method for iodide detection using diameter-based measurement on laminated paper-based analytical device (LPAD). Our LPAD is simplified fabricated with no required hydrophobic barrier for patterning a detection flow channel. In the absence of a flow channel, the liquid is imbibed radially into the filter paper outward from the small center inlet hole. This imbibition leads to the measurement of diameter after the formation of the precipitation product. For LPAD, a rectangular piece of Whatman filter paper is cut and positioned between a top laminating sheet, which has a hole punched for inlet, and a bottom laminating sheet. The central inlet hole enables access to the liquid on the paper. For iodide analysis, silver nitrate is dispensed first, followed by the iodide standard/sample. This process leads to the formation of pale-yellow silver iodide solids on the paper. The chromate indicator solution is subsequently loaded to the paper. The color of the chromate solution imparts a tint to the circular band of silver iodide solids, resulting in an intense yellow color. When the silver iodide precipitates are stained, the chromate solution typically serves as an indicator, chemically reacting to an excess silver nitrate solution moistened on paper. This reaction generates an outer reddish-brown ring of silver chromate. The distinct color contrast between the intense yellow (tinted silver iodide) and reddish-brown (silver chromate) allows for clear observation of the measurement boundary, facilitating diameter-based measurements of the circular band of silver iodide. We demonstrate the applicability in analysis of pharmaceutical KI tablets. The quantitative results obtained from diameter-based measurement LPAD show good agreement with those obtained from the potentiometric method. Our hydrophobic barrier-free LPAD is rapid, cost-effective, low liquid consumption and applicable for onsite analysis.

这项工作首次在层压纸基分析装置(LPAD)上利用基于直径测量的精氨酸莫尔法检测碘化物。我们的 LPAD 制作简单,不需要疏水屏障来图案化检测流道。在没有流道的情况下,液体从中间的小进水孔向外径向浸入滤纸。沉淀产物形成后,通过这种浸入方式测量直径。在 LPAD 中,一张长方形的 Whatman 滤纸被切割并放置在顶部覆膜板和底部覆膜板之间。中央的进气孔可以让液体进入滤纸。在进行碘化物分析时,首先分配硝酸银,然后分配碘化物标准品/样品。这一过程会在试纸上形成淡黄色的碘化银固体。随后将铬酸盐指示剂溶液加载到试纸上。铬酸盐溶液的颜色会使碘化银固体的环形带着色,从而形成浓黄色。当碘化银沉淀被染色时,铬酸盐溶液通常作为指示剂,与湿润在纸上的过量硝酸银溶液发生化学反应。这种反应会在外部生成一个红棕色的铬酸银环。浓黄色(着色碘化银)和红棕色(铬酸银)之间明显的颜色对比可以清楚地观察到测量边界,便于对碘化银环带进行基于直径的测量。我们在分析药用碘化钾片时演示了这种方法的适用性。基于直径测量的 LPAD 所获得的定量结果与电位计方法所获得的结果非常一致。我们的疏水无屏障 LPAD 方法快速、经济、耗液少,适用于现场分析。
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引用次数: 0
Purity Determination of High-Purity BeO using Gravimetric Analysis with Stepwise Conversions of Weighing Form of Be 利用重量分析法分步转换 Be 的称量形式测定高纯度 BeO 的纯度
Q3 Chemistry Pub Date : 2024-02-28 DOI: 10.1016/j.talo.2024.100307
Tsutomu Miura

Ensuring accurate and precise measurements of analytes is crucial in gravimetric analysis but can be jeopardized by overlooking analyte in the filtrate, washing solutions, residue during analytical procedure, and weighing form of analyte. To address this issue, a stoichiometric approach for the purity determination of high-purity BeO was developed using gravimetric analysis. This method involved the systematic stoichiometric evaluation of the stepwise conversion of the different weighing forms of Be. The initial BeO sample underwent sequential conversion to the Be complex with N-benzoyl-N-phenyl hydroxylamine, BeO, and BeSO4; each was evaluated for stoichiometry via gravimetric analysis. The amounts of Be in the filtrate and washing solution, and their loss during sample preparation were measured using inductively coupled plasma-optical emission spectrometry (ICP-OES). Additionally, the sulfate content in the BeSO4 precipitate was determined by the gravimetric analysis of BaSO4 to validate the stoichiometry of the precipitate. The observed mass ratios of different Be weighing forms were compared with the theoretical values at each conversion step. The composition of the final converted BeSO4, i.e., the observed mass ratio of BaSO4/BeSO4, was comparable to the theoretical composition. The purity of the initial BeO sample was determined to be 100.03 % ± 0.17 % (k=2). The measured purity was successfully validated by comparing it with that of NIST SRM 3105a Be standard solution using ICP-OES.

在重量分析中,确保准确和精确测量分析物至关重要,但忽略滤液中的分析物、洗涤溶液、分析过程中的残留物以及分析物的称量形式可能会影响测量结果。为解决这一问题,我们开发了一种利用重量分析法测定高纯度 BeO 纯度的化学计量法。该方法涉及对不同称量形式的 Be 的逐步转换进行系统的化学计量学评估。最初的 BeO 样品依次转化为与 N-苯甲酰基-N-苯基羟胺、BeO 和 BeSO4 的 Be 复合物;每种复合物都通过重量分析法进行了化学计量学评估。使用电感耦合等离子体-光发射光谱法(ICP-OES)测量了滤液和洗涤液中的铍含量及其在样品制备过程中的损失。此外,BeSO4 沉淀中的硫酸盐含量是通过 BaSO4 重量分析法确定的,以验证沉淀的化学计量学。在每个转化步骤中,将观察到的不同 Be 称重形式的质量比与理论值进行比较。最终转化的 BeSO4 的成分,即观察到的 BaSO4/BeSO4 的质量比,与理论成分相当。经测定,初始 BeO 样品的纯度为 100.03 % ± 0.17 %(k=2)。通过使用 ICP-OES 与 NIST SRM 3105a Be 标准溶液进行比较,成功验证了所测得的纯度。
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引用次数: 0
Upconverting nanophosphors for various sensing applications 用于各种传感应用的上转换纳米磷化物
Q3 Chemistry Pub Date : 2024-02-28 DOI: 10.1016/j.talo.2024.100302
Jitender Kumar, Indrajit Roy

Background

Upconverting nanophosphors (UCNPs) are composed of certain nanosized inorganic host materials doped with specific rare earth ions. They exhibit the phenomenon of photon upconversion, whereby the sequential absorption of two or more low energy photons is followed by luminescent emission of multiple photons with higher energy. In addition, they have other attractive optical features such as NIR-absorption, sharp emission bands, exceptional photostability and high quantum yield. These properties make UCNPs lucrative probes for number of optical applications, such as imaging, sensing and theranostics.

Overview

UCNPs have been used as specific and ultrasensitive probes for sensing of variety of analytes, such as gas molecules, metal ions, fine particles, pH and a number of molecules, biochemicals and macromolecules. Their absorption and emission in the NIR range facilitates the background free biosensing in situ, in vitro and in vivo. Their multicoloured emissions allow a suitable emission band to serve as a donor for energy transfer to another attached optical probe, while a separate emission band to serve as an internal reference in ratiometric sensing applications. This comprehensive review provides an in-depth exploration of the versatile applications of UCNPs in the optical sensing of various analytes based on energy transfer mechanisms, with emphasis on the recent developments. . The focus is on synthesizing UCNPs, surface functionalization, and their effective utilization especially in detecting gas molecules, pH changes, ions, free radicals, and disease biomarkers. Through a meticulous examination of the use of UCNPs, this review aims to contribute to a deeper understanding of their potential and pave the way for future advancements and challenges in optical sensing.

背景上转换纳米磷酸盐(UCNPs)由掺杂特定稀土离子的某些纳米级无机主材料组成。它们表现出光子上转换现象,即在连续吸收两个或多个低能光子后,发出多个高能光子。此外,它们还具有其他诱人的光学特性,如近红外吸收、尖锐的发射带、优异的光稳定性和高量子产率。这些特性使 UCNPs 成为成像、传感和治疗等多种光学应用的有利探针。概述UCNPs 已被用作特异性超灵敏探针,用于传感各种分析物,如气体分子、金属离子、微粒、pH 值以及多种分子、生化物质和大分子。它们在近红外范围内的吸收和发射有助于在原位、体外和体内进行无背景生物传感。它们的多色发射允许一个合适的发射波段作为供体将能量传递给另一个附着的光学探针,而一个单独的发射波段则可作为比率测量传感应用中的内部参考。本综述深入探讨了 UCNPs 在基于能量转移机制的各种分析物光学传感中的广泛应用,并重点介绍了最近的发展情况。.重点是 UCNPs 的合成、表面功能化及其有效利用,尤其是在检测气体分子、pH 值变化、离子、自由基和疾病生物标记物方面。通过对 UCNPs 用途的细致研究,本综述旨在加深对其潜力的理解,并为未来光学传感领域的进步和挑战铺平道路。
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引用次数: 0
Developing a prediction method for physicochemical characteristics of Pontianak Siam orange (Citrus suhuiensis cv. Pontianak) based on combined reflectance-Fluorescence spectroscopy and artificial neural network 基于反射荧光光谱和人工神经网络的坤甸暹罗橙(Citrus suhuiensis cv. Pontianak)理化特性预测方法的开发
Q3 Chemistry Pub Date : 2024-02-27 DOI: 10.1016/j.talo.2024.100303
Sandra , Abdullah Said , Ahmad Avatar Tulsi , Dina Wahyu Indriani , Rini Yulianingsih , La Choviya Hawa , Naoshi Kondo , Dimas Firmanda Al Riza

The slightly sweet and acidic taste offered by Pontianak Siam oranges is influenced by the total soluble solids (TSS) and acidity in the fruit, in which, measuring these attributes is commonly performed using instruments that potentially damage the fruit's structure, thus, impractical for fresh fruit products. Moreover, the process of classifying the quality of fresh oranges has been based on physical appearance, leading to subjective results. Correspondingly, the objective of the study is to develop a prediction method for the physicochemical characteristics of Pontianak Siam oranges based on VIS-NIR-Fluorescence spectroscopy and an artificial neural network (ANN) model. The method is applicable to classify oranges based on physicochemical characteristics without damaging the fruit's structure. As a result, the best model for classifying the maturity level of Pontianak Siam oranges was obtained using a dataset with all feature combined spectra, attaining a training accuracy of 0.99 and testing accuracy of 1. The best model for predicting TSS was obtained using all feature combined spectra dataset, attaining R2 training = 0.89 and R2 testing = 0.91. The best model for predicting acidity was obtained using all feature reflectance spectra datasets, attaining R2 training = 0.96 and R2 testing = 0.97. The best model for predicting fruit firmness was obtained using all feature reflectance spectra dataset, attaining R2 training = 0.97, R2 testing = 0.89. Overall, the combination of Vis-NIR reflectance and fluorescence spectroscopy have the potential to be applied for non-destructive assessment of citrus quality in terms of visual classification and maturity parameters prediction.

坤甸暹罗橙的微甜和微酸口感受水果中总可溶性固形物(TSS)和酸度的影响,而测量这些属性通常使用可能会破坏水果结构的仪器,因此对新鲜水果产品来说并不实用。此外,新鲜橙子的质量分级过程一直以物理外观为基础,导致主观结果。因此,本研究旨在开发一种基于 VIS-NIR 荧光光谱和人工神经网络(ANN)模型的暹罗椪柑理化特性预测方法。该方法适用于在不破坏水果结构的情况下根据理化特性对橙子进行分类。结果,使用所有特征组合光谱数据集获得了坤甸暹罗橙成熟度分类的最佳模型,训练精度为 0.99,测试精度为 1。使用所有特征反射光谱数据集获得了预测酸度的最佳模型,训练 R2 = 0.96,测试 R2 = 0.97。使用所有特征反射光谱数据集获得了预测果实硬度的最佳模型,训练 R2 = 0.97,测试 R2 = 0.89。总之,可见光-近红外反射光谱和荧光光谱的结合有望在视觉分类和成熟度参数预测方面应用于柑橘质量的非破坏性评估。
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引用次数: 0
Future advances of artificial biosensor technology in biomedical applications 人工生物传感器技术在生物医学应用中的未来发展
Q3 Chemistry Pub Date : 2024-02-27 DOI: 10.1016/j.talo.2024.100301
Smriti Gaba , Nidhi Chauhan , Ramesh Chandra , Utkarsh Jain

Recent advancements in synthetic biology have facilitated the concept of a cell-based and cell-free biosensing platform, which enables the identification of molecular signals encompassing metal/chemical to disease biomarkers. The artificial sensing incorporates the concept of both whole-cell and cell-free biosensing strategies, which include highly regulated natural and synthetic components to exhibit genetically encoded molecular sensing properties. These sensors utilize protein expression to release signalling molecules as the result of received input to facilitate the detection of analytes. Intending to use modified living cells or artificial cells in biosensing, the proposed study highlights the importance of cell-based and cell-free sensors in biomedical and diagnostics. The article's first section will explain the biosensing types including cell-free, cell-based, vesicle-based, and paper-based sensing where sensing relies on cell, cellular components, and cell-free systems which mostly involve transcriptional or translational machinery. It highlights the advantages, disadvantages, and challenges of advancing approaches. The second section of the article elaborates on the principle of sensing and the strategies involved. Though very few studies have been reported on this topic, therefore, the current article focuses on the artificial sensors that have been designed for medical and diagnostic purposes. The review also marks the current and future advancements in the field including artificial intelligence, nanotechnology, stem cells, and omics. Sensing recently has a big impact on disease diagnosis as well as drug development and targeted therapies. While newly developed biology-based diagnostics technologies still have high costs, require highly trained personnel, suffer stability issues and reduce sensor performance. Therefore, this review brings readers’ attention to advances and challenges in the following field and promotes the resolution of medical and diagnostics issues in the future.

合成生物学的最新进展促进了基于细胞和无细胞生物传感平台概念的发展,该平台能够识别从金属/化学到疾病生物标志物的分子信号。人工传感结合了全细胞和无细胞生物传感策略的概念,其中包括高度调节的天然和合成成分,以显示基因编码的分子传感特性。这些传感器利用蛋白质表达来释放信号分子,作为接收输入的结果,以促进分析物的检测。拟议的研究打算在生物传感中使用经修饰的活细胞或人工细胞,这凸显了基于细胞和无细胞传感器在生物医学和诊断学中的重要性。文章的第一部分将解释生物传感的类型,包括无细胞传感、细胞传感、囊泡传感和纸质传感,其中传感依赖于细胞、细胞成分和无细胞系统,这些系统大多涉及转录或翻译机制。文章强调了这些先进方法的优缺点和挑战。文章的第二部分阐述了传感原理和相关策略。虽然有关这一主题的研究报告很少,但本文重点讨论了为医疗和诊断目的而设计的人工传感器。综述还介绍了该领域当前和未来的进展,包括人工智能、纳米技术、干细胞和 Omics。最近,传感技术对疾病诊断、药物开发和靶向治疗产生了重大影响。然而,新开发的基于生物学的诊断技术仍存在成本高、需要训练有素的人员、稳定性问题以及传感器性能降低等问题。因此,这篇综述让读者关注以下领域的进展和挑战,促进未来医疗和诊断问题的解决。
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引用次数: 0
Two-step dual-layer SPE method to separate antibacterial and antioxidant mushroom compounds 两步双层固相萃取法分离抗菌和抗氧化蘑菇化合物
Q3 Chemistry Pub Date : 2024-02-27 DOI: 10.1016/j.talo.2024.100304
Dániel Krüzselyi, Péter G. Ott, Ágnes M. Móricz

We developed a two-step dual-layer solid-phase extraction (SPE) method to separate and concentrate minor bioactive compounds of golden oyster mushroom (Pleurotus citrinopileatus) methanol extract. The mushroom extract is rich in fatty acids, including the highly abundant (20–35 %) and antibacterial linoleic acid. To eliminate the masking effect of linoleic acid in the bioassay, its removal was achieved by homemade dual-layer SPE that consisted of a lower C18 phase and an upper silica gel phase with the dried-on extract. In the first step the elution was performed with water (aqueous eluate), and then the dried C18 and silica gel phases were eluted separately with methanol (C18 methanol eluate and silica gel methanol eluate, respectively). Thin-layer chromatography (TLC)-effect-directed analysis demonstrated that the aqueous eluate contained sugars and antioxidants (TLC-DPPH); fatty acids were present in the silica gel methanol eluate, in the C18 methanol eluate the biologically active minor secondary metabolites were concentrated (TLC–Bacillus subtilis assay) beside 2 % remained linoleic acid according to HPLC-UV measurement. As a result, the antioxidant and antibacterial metabolites divided, and the low-abundant antibacterial compounds could be detected. Based on this study, the substance found at RF 0.25 by TLC-DPPH played the most significant role in generating the antioxidant effect of the aqueous eluate, equivalent to 5.07 ± 0.09 µg of gallic acid. Moreover, each SPE fraction and raw extract were tested for antibacterial effect using a microplate assay with a non-pathogenic Gram-positive Bacillus subtilis. The P. citrinopileatus extract (MIC-value – 50 µg/mL) and C18 methanol eluate (MIC-value - 25 µg/mL) inhibited the growth of B. subtilis while the other eluates did not.

我们开发了一种两步双层固相萃取(SPE)法,用于分离和浓缩金杏鲍菇甲醇提取物中的次要生物活性化合物。杏鲍菇提取物中含有丰富的脂肪酸,包括含量极高(20-35%)的抗菌亚油酸。为了消除亚油酸在生物测定中的掩蔽效应,采用了自制的双层固相萃取法来去除亚油酸。第一步用水洗脱(水洗脱液),然后用甲醇分别洗脱干燥的 C18 相和硅胶相(分别为 C18 甲醇洗脱液和硅胶甲醇洗脱液)。薄层色谱(TLC)-效应定向分析表明,水洗脱液中含有糖类和抗氧化剂(TLC-DPPH);硅胶甲醇洗脱液中含有脂肪酸,C18 甲醇洗脱液中浓缩了具有生物活性的次生代谢物(TLC-枯草杆菌测定),HPLC-UV 测定结果显示,亚油酸残留量为 2%。结果,抗氧化和抗菌代谢物被分开,低含量的抗菌化合物被检测到。根据这项研究,TLC-DPPH 在 RF 值为 0.25 时发现的物质对水洗脱液的抗氧化作用最为显著,相当于 5.07 ± 0.09 µg 没食子酸。此外,每种固相萃取物馏分和提取物原液都通过微孔板法与非致病性革兰氏阳性枯草杆菌进行了抗菌效果测试。柠檬皮提取物(MIC 值 - 50 µg/mL)和 C18 甲醇洗脱液(MIC 值 - 25 µg/mL)抑制了枯草杆菌的生长,而其他洗脱液则没有抑制作用。
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引用次数: 0
Fine-tuning of highly bright benzo[c,d]indole-oxazolopyridine cyanine dye for nucleolar RNA imaging in living cells 微调高亮度苯并[c,d]吲哚-噁唑吡啶青染料,用于活细胞核 RNA 成像
Q3 Chemistry Pub Date : 2024-02-27 DOI: 10.1016/j.talo.2024.100308
Nao Togashi, Masaaki Nagaoka, Kei Higuchi, Yukina Yoshino, Yawen Wu, Yusuke Sato, Seiichi Nishizawa

Here we report on fine-tuning of highly bright fluorogenic cyanine dye, benzo[c,d]indole-oxazolo[5,4-c]pyridine (BIOP: λem = 570 nm, Φfree = 0.00038, Φbound = 0.52), recently developed by our group for nucleolar RNA imaging in living cells. We tuned an emission maximum to the longer wavelength by replacing oxazolopyridine unit with its isomer. The resulting probe with oxazolo[4,5-b]pyridine, named BIOP [4,5-b], exhibited a significant off-on signaling ability for RNA (λem = 580 nm, Φfree = 0.002, Φbound = 0.46) that almost compared with BIOP. BIOP [4,5-b] was applicable to live-cell imaging, where wash-free protocol was available. Importantly, the slight change in spectral features would be expected to minimize the false signal from BIOP [4,5-b] in co-staining experiments with typical green-emissive dyes. In addition, thanks to the unique spectral shape that is typical of cyanine dyes, we demonstrate that yellow-emissive BIOP [4,5-b] also did work as a pseudo red-emissive dye (λem > 600 nm) for live-cell RNA imaging, for which we just switch filter set to typical one for real red-emissive dyes (Ex 560/40 nm; Em 645/75 nm).

在此,我们报告了对高亮度荧光青染料苯并[]吲哚-噁唑并[5,4-]吡啶(BIOP: = 570 nm, Φ = 0.00038, Φ = 0.52)的微调情况,该染料是我们小组最近开发的用于活细胞核 RNA 成像的染料。我们用其异构体取代了噁唑并吡啶单元,从而将发射最大值调到了更长的波长。由此产生的含有噁唑并[4,5-]吡啶的探针被命名为 BIOP [4,5-],它对 RNA(= 580 nm, Φ = 0.002, Φ = 0.46)具有显著的关断信号能力,与 BIOP 几乎不相上下。BIOP[4,5-]适用于免洗方案的活细胞成像。重要的是,光谱特征的细微变化有望将 BIOP [4,5-] 在与典型的绿色发射染料共染实验中产生的假信号降至最低。此外,得益于氰基染料典型的独特光谱形状,我们证明黄色亲和性 BIOP [4,5-] 也可以作为伪红色亲和性染料(> 600 nm)用于活细胞 RNA 成像,为此我们只需将滤光片设置切换为真正红色亲和性染料的典型滤光片(Ex 560/40 nm; Em 645/75 nm)。
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