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A dual enzyme-mimicking nanoplatform based on CoMn-LDHs for ultrasensitive detection of L-cysteine: Toward environmental and food safety monitoring 基于con - ldhs的双酶模拟纳米平台超灵敏检测l -半胱氨酸:用于环境和食品安全监测
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-11-17 DOI: 10.1016/j.talo.2025.100596
Salma Shalaby , Jenny Jeehan Mohamed Nasr , Asmaa Kamal El-Deen
Cysteine (CYS) is a pivotal biomolecule involved in both biological and environmental processes, yet conventional detection methods suffer from low sensitivity, complex procedures, and interference issues. In this study, we report the development of cobalt-manganese layered double hydroxides (CoMn-LDHs) as a novel nanomaterial-based sensing platform with dual oxidase/peroxidase mimicry for label-free CYS detection. The synthesized CoMn-LDHs exhibit exceptional catalytic activity, efficiently oxidizing the chromogenic substrate TMB through the generation of reactive species such as superoxide anion (O₂•⁻) and hydroxyl radicals (•OH), which significantly enhance sensitivity and selectivity. This dual-enzyme mimic mechanism enables a rapid, colorimetric detection method with ultra-low detection limits of 0.024 μM and 0.12 μM for oxidase and peroxidase modes, respectively, outperforming traditional techniques. The sensor demonstrates excellent performance in real samples, including mineral water, tap water, and milk, with high recovery rates (> 98 %) and low relative standard deviation (%RSD < 2 %), showcasing its robust stability and reproducibility in complex matrices. This work presents a cost-effective, highly sensitive, and selective platform for CYS detection, offering significant advancements in environmental monitoring and food safety applications through the innovative use of nanomaterial-based enzyme mimics.
半胱氨酸(CYS)是一种参与生物和环境过程的关键生物分子,但传统的检测方法存在灵敏度低、程序复杂和干扰问题。在这项研究中,我们报道了钴锰层状双氢氧化物(com - ldhs)的发展,作为一种新型的基于纳米材料的传感平台,具有双氧化酶/过氧化物酶模拟,用于无标记CYS检测。合成的com - ldhs具有特殊的催化活性,通过生成超氧阴离子(O₂•毒血症)和羟基自由基(•OH)等活性物质,有效地氧化显色底物TMB,显着提高了灵敏度和选择性。该双酶模拟机制可实现氧化酶和过氧化物酶模式的快速比色检测,超低检出限分别为0.024 μM和0.12 μM,优于传统技术。该传感器在矿泉水、自来水和牛奶等实际样品中表现出优异的性能,具有高回收率(98%)和低相对标准偏差(%RSD < 2%),在复杂基质中具有良好的稳定性和重复性。这项工作提出了一个具有成本效益,高灵敏度和选择性的CYS检测平台,通过创新地使用基于纳米材料的酶模拟物,在环境监测和食品安全应用方面取得了重大进展。
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引用次数: 0
Biomimetic 3D printable electrochemical filament integrated with CuO nanopowder for enzyme-free biosensing 集成CuO纳米粉末的仿生3D打印电化学丝用于无酶生物传感
IF 4.1 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-05-24 DOI: 10.1016/j.talo.2025.100480
Leonidas Papagiannakopoulos , Vasiliki Polyzopoulou , Loizos Tsolakis , Eleni Sorkou, Eleni Koukouviti, Anastasios Economou, Christos Kokkinos
3D printing technology enables the on-demand fabrication of low-cost thermoplastic electrodes, which have shown promising results in enzymatic bioassays. To fully harness the potential of 3D printing in electrochemical biosensing, this work introduces a new generation of tailor-made conductive filament integrated with a biomimetic functional material for the 3D printing of ready-to-use sensors designed for enzyme-free biodeterminations. The filament was synthesized using the solvent casting method and was composed of polylactic acid as the base, carbon black as the conductive filler, and CuO nanopowder as the artificial nanozyme. The filament was characterized using thermogravimetry, energy-dispersive X-ray spectroscopy, scanning electron microscopy, and electrochemical techniques. The as-printed sensors enabled direct electrochemical monitoring of crucial bioindicators—glucose (GLU), lactic acid (LA), and creatinine (CRE)—with exceptional sensitivity, offering limits of detection of 5.1 µM, 0.12 mM, and 1.5 µM, respectively. Moreover, the sensors exhibited high anti-interference capability and were successfully applied to the determination of the target biomarkers in blood, sweat, and urine samples. The amperometric determination of GLU was based on the conversion of integrated CuO to CuOOH, which subsequently oxidized GLU to gluconic acid. The voltammetric determination of CRE was based on the formation of copper-creatinine complexes, resulting in the suppression of the oxidation signal of electrogenerated copper. Meanwhile, LA voltammetric detection relied on the characteristic increases in the redox signals of CuO. This work paves the way for the development of accessible, point-of-need printable, and ready-to-use electrochemical biomimetic sensors for easy applications in the biosensing field.
3D打印技术能够按需制造低成本的热塑性电极,在酶促生物分析中显示出有希望的结果。为了充分利用3D打印在电化学生物传感方面的潜力,这项工作引入了新一代定制的导电丝和仿生功能材料,用于3D打印用于无酶生物测定的即用型传感器。以聚乳酸为基料,炭黑为导电填料,纳米氧化铜粉为人工纳米酶,采用溶剂浇铸法合成该长丝。利用热重法、能量色散x射线光谱学、扫描电子显微镜和电化学技术对灯丝进行了表征。打印的传感器能够直接电化学监测关键的生物指标-葡萄糖(GLU),乳酸(LA)和肌酐(CRE) -具有卓越的灵敏度,分别提供5.1µM, 0.12 mM和1.5µM的检测限。此外,该传感器表现出高抗干扰能力,并成功应用于血液、汗液和尿液样本中目标生物标志物的测定。GLU的安培测定是基于整合的CuO转化为CuOOH, CuOOH随后将GLU氧化为葡萄糖酸。伏安法测定CRE是基于铜-肌酐络合物的形成,从而抑制电生铜的氧化信号。同时,LA伏安检测依赖于CuO氧化还原信号的特征增加。这项工作为开发可访问的、可打印的、即用型电化学仿生传感器铺平了道路,使其易于在生物传感领域应用。
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引用次数: 0
A Novel and Green UFLC-MS/MS Method for Quantification of Amantadine and Levodopa in Polymeric Nanoparticles: Application to determine Drug loading (%DL), Drug entrapment (%DEE) and Drug release profile 一种新型、绿色的UFLC-MS/MS定量纳米聚合物中金刚烷胺和左旋多巴的方法:用于测定载药量(%DL)、包裹度(%DEE)和药物释放谱
IF 4.1 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-05-21 DOI: 10.1016/j.talo.2025.100476
Akanksha Lahiri , Balamuralidhara V , Hemanth Vikram PR , Karthika Paul

Introduction

Levodopa, a dopamine precursor, widely prescribed drug in Parkinson’s disease management possesses a side effect of Levodopa-induced dyskinesia (LID). Amantadine hydrochloride, an NMDA receptor antagonist is co-encapsulated along with levodopa and formulates as polymeric nanoparticles (NPs) so as to overcome the side effects and act synergistically to enhance therapeutic outcome.

Methodology

In current work, we developed a novel, green Ultra-Fast Liquid Chromatography-Tandem Mass Spectrometry (UFLC-MS/MS) technique for the simultaneous quantification of amantadine and levodopa in polymeric nanoparticles. A triple quadrupole analyser with a multiple reaction monitoring (MRM) scan mode and an atmospheric pressure chemical ionization (APCI) source. A Waters Symmetry C8 column (150 × 4.6 mm, 3.5 μm) maintained at 40 °C was used for the chromatographic separation. In order to ensure sensitive and specific analyte detection, the mobile phase consisted of 0.1 % formic acid in water and methanol (40:60) with a total run time of 5 min. Excellent linearity, recovery, accuracy, and sensitivity were validated by method validation. Greenness assessment was done by AGREE, GAPI and AES metrics.

Results

Proposed green UFLC-MS/MS method effectively quantifies Levodopa and Amantadine in polymeric nanoparticles, followed by accurate evaluation of %DL, %DEE, and drug release profiles. %DEE values were observed for Levodopa (89.6%) and Amantadine (90.16 %), with corresponding %DL values of 20.5% and 24.10%, indicating substantial drug loading capacity. The in vitro drug release profiles demonstrated sustained release behaviour, with 91.89% ± 0.362 of Levodopa and 92% ± 0.362 of Amantadine released over the study period.

Conclusion

Using GAPI, AGREE, and AES criteria, the created analytical method's greenness was carefully assessed. The outcomes were then compared to previously published methods in the literature. For the simultaneous measurement of amantadine and levodopa in nanoparticles, our innovative UFLC-MS/MS technology offered a dependable and extremely sensitive method.
左旋多巴是一种多巴胺前体,是帕金森病治疗中广泛使用的处方药,它具有左旋多巴诱导的运动障碍(LID)的副作用。盐酸金刚烷胺是一种NMDA受体拮抗剂,与左旋多巴共包被,形成聚合物纳米颗粒(NPs),以克服副作用并协同作用,提高治疗效果。在目前的工作中,我们开发了一种新型的绿色超快速液相色谱-串联质谱(UFLC-MS/MS)技术,用于同时定量聚合物纳米颗粒中的金刚烷胺和左旋多巴。具有多反应监测(MRM)扫描模式和大气压化学电离(APCI)源的三重四极杆分析仪。色谱柱为Waters Symmetry C8柱(150 × 4.6 mm, 3.5 μm),保持温度为40℃。为确保分析物检测的敏感性和特异性,流动相为0.1%甲酸水溶液和甲醇(40:60),总运行时间为5 min。通过方法验证验证了良好的线性、回收率、准确性和灵敏度。绿色评估采用AGREE、GAPI和AES指标。结果建立的绿色UFLC-MS/MS方法能有效地定量左旋多巴和金刚烷胺在聚合物纳米颗粒中的含量,并能准确评价其%DL、%DEE和药物释放谱。左旋多巴(89.6%)和金刚烷胺(90.16%)的DEE值为%,相应的DL值分别为20.5%和24.10%,说明其载药量较大。体外释药表现为缓释行为,左旋多巴释药率为91.89%±0.362%,金刚烷胺释药率为92%±0.362%。结论采用GAPI、AGREE和AES标准对所建立的分析方法进行了绿色度评价。然后将结果与文献中先前发表的方法进行比较。对于纳米颗粒中金刚烷胺和左旋多巴的同时测定,我们创新的UFLC-MS/MS技术提供了一种可靠且灵敏度极高的方法。
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引用次数: 0
Developments of compact, energy-efficient thin-film-based multilayered oxygen sensors 紧凑、节能的薄膜多层氧传感器的发展
IF 4.1 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-05-25 DOI: 10.1016/j.talo.2025.100479
Nair Afijith Ravindranath , V. Jayaraman , K.I. Gnanasekar
This short review is intended to provide an overview of the recent developments in the field of multilayered thin films of solid electrolyte-based electrochemical oxygen sensors. This work reviews the factors that prevent the realization of multilayer-based thin film electrochemical sensors and outlines the methodologies to be adopted to circumvent them. It demonstrates the feasibility of energy-efficient compact sensors as they offer low thermal mass and ease of integration with the thin film heater for operation. Design of a multilayered sensor, deposition of high-quality thin films of various components of electrochemical oxygen sensing device, density of YSZ thin film electrolyte, interdiffusion of components and their impact on electrical property, long-term stability of thin layered device and its operation at lower temperatures of about 623 to 673 K as against 923 K for bulk devices have been addressed.
本文综述了固体电解质基多层薄膜电化学氧传感器领域的最新进展。这项工作回顾了阻碍实现多层薄膜电化学传感器的因素,并概述了采用的方法来绕过它们。它证明了节能紧凑型传感器的可行性,因为它们提供低热质量和易于与薄膜加热器集成操作。本文讨论了多层传感器的设计、电化学氧传感器件各组分高质量薄膜的沉积、YSZ薄膜电解质的密度、各组分间的相互扩散及其对电性能的影响、薄层器件的长期稳定性及其在623 ~ 673 K(较低温度为923 K)下的运行。
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引用次数: 0
Quantification of phytochemicals of different parts of Oroxylum indicum (L.) Kurz for the evidence-based substitution of the official part (root) with aerial part 籼稻(Oroxylum indicum)不同部位植物化学成分的定量分析Kurz为官方部分(根)与空中部分的循证替代
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-08-05 DOI: 10.1016/j.talo.2025.100528
Megha Nigam , Yashika Gandhi , Vijay Kumar , Hemant Soni , Rishi Kumar Saxena
Oroxylum indicum (L.) Kurz (OI) or Shyonaka or Sona Patha is an endangered medicinal plant used in various traditional medicines. The roots of OI are the most commonly used part in preparing numerous traditional medicines worldwide. To conserve the environment, it is necessary to check the substitution of official parts with aerial parts like leaf and stem bark using sophisticated techniques. The present study developed HPLC and ICP-OES methods to quantify OI's main phytoconstituents and metal ions in four parts (root, root bark, stem bark, and leaf). The phytochemicals viz. vanillic acid, trans-ferulic acid, baicalein, and chrysin were quantified by HPLC study, where the significantly better amounts of baicalein and chrysin (baicalein > chrysin) were quantified in aerial parts (leaf and stem bark) as compared to root and root bark. ICP-OES elemental analysis has revealed that all parts of OI are good sources of Ca, K, Na, Mg, Mn, Fe, and Zn. GC–MS study was performed to identify the volatile compounds of all parts of OI. In antioxidant studies (TPC, TFC, and DPPH assays), the leaf showed a better IC50 value, followed by stem bark, root bark, and root. In the antacid study, the leaf has shown better activity, followed by root bark, stem bark, and root. In the anti-bacterial assay, all parts of OI significantly inhibited Staphylococcus aureus and Salmonella typhi strains, where root bark and leaf demonstrated improved activities. In computational studies, the invitro antacid, antioxidant, and anti-bacterial activities were confirmed, where most of the phytochemicals demonstrated binding energies over standard drugs. Overall, the study revealed that all parts of OI, including roots and aerial parts, might be medicinally useful, and leaves may be used as a nutritional food. Moreover, the official part, i.e., root or root bark used in traditional medicines, may be replaced with aerial parts (leaf or stem bark) to conserve the environment. After the in-depth pharmacological and toxicological studies, all parts of OI Oroxylum indicum might be incorporated into pharmaceutics.
籼稻(Oroxylum indicum)Kurz (OI)或Shyonaka或Sona Patha是一种濒危的药用植物,用于各种传统药物中。成骨不全症的根源是世界范围内许多传统药物制备中最常用的部分。为了保护环境,有必要使用复杂的技术检查用树叶和茎皮等空中部分代替官方部分。本研究采用HPLC和ICP-OES两种方法定量测定了黄芪根、根皮、茎皮和叶四个部分的主要植物成分和金属离子。采用高效液相色谱法测定了植物化学物质香草酸、反式阿魏酸、黄芩素和菊花素的含量,其中黄芩素和菊花素的含量显著高于黄芩素和菊花素;与根和根皮相比,在地上部分(叶和茎皮)中量化了菊花素。ICP-OES元素分析表明,OI的所有部分都是Ca, K, Na, Mg, Mn, Fe和Zn的良好来源。采用气相色谱-质谱法对成骨成骨各部位的挥发性化合物进行鉴定。在抗氧化研究(TPC, TFC和DPPH测定)中,叶显示出较好的IC50值,其次是茎皮,根皮和根。在抗酸研究中,叶表现出较好的抗酸活性,其次是根皮、茎皮和根。在抗菌试验中,OI的所有部分都能显著抑制金黄色葡萄球菌和伤寒沙门氏菌菌株,其中根皮和叶子的活性有所提高。在计算研究中,体外抗酸、抗氧化和抗菌活性得到证实,其中大多数植物化学物质比标准药物表现出结合能。总的来说,这项研究揭示了成骨不全的所有部分,包括根部和地上部分,可能都有药用价值,叶子可能被用作营养食品。此外,官方部分,即传统药物中使用的根或根皮,可以用空气部分(叶或茎皮)代替,以保护环境。经过深入的药理学和毒理学研究,该植物的各个部位都有可能被纳入到药物中。
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引用次数: 0
Accuracy assessment of a micro-Raman spectroscopy method for small microplastic particles in infant milk formula 微拉曼光谱法测定婴儿配方奶粉中塑料微粒的准确性评价
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-10-31 DOI: 10.1016/j.talo.2025.100586
Mara Putzu , Marta Barbaresi , Marta Fadda , Alessio Sacco , Maurizio Piergiovanni , Matteo Masino , Federica Bianchi , Korinna Altmann , Nizar Benismail , Laureen Coïc , Ivana Fenoglio , Monica Mattarozzi , Andrea Mario Rossi , Maria Careri , Andrea Mario Giovannozzi
The presence of microplastics (MPs) in the food chain is increasingly documented, raising concerns over potential risks to human health. Despite growing efforts, standardized methods for MPs detection in food matrices remain limited. This study presents an interlaboratory comparison (ILC) aimed at assessing the accuracy and comparability of an analytical approach for the identification and quantification of small MPs (5–100 µm) in infant milk powder using µ-Raman spectroscopy and a representative polyethylene terephthalate (PET) reference material (RM). The RM, formulated as water-soluble tablets, was designed to replicate the morphology, size distribution, and polymer composition of environmentally relevant MPs, and was previously assessed for homogeneity and stability for mass fraction and particle numbers.
The approach was assessed using two PET RM batches with different MPs particle numbers (high load batch: 1759 ± 141 MPs; low load batch: 160 ± 22 MPs), subjected to an enzymatic–chemical digestion, followed by µ-Raman analysis performed independently in two laboratories with different instruments and operators. Results are reported as absolute particle counts per analyzed sample and demonstrated excellent recovery across all size classes, including the smallest particles (down to 5 µm), with recovery rates ranging from 82 % to 88 %, in good agreement with the RM reference values.
The analytical approach proved to be robust, reproducible, and suitable for low-level MPs quantification in complex food matrices, supporting ongoing efforts toward method harmonization and standardization for reliable MPs monitoring in the food sector.
微塑料(MPs)在食物链中的存在被越来越多地记录下来,引发了人们对人类健康潜在风险的担忧。尽管越来越多的努力,在食品基质中检测MPs的标准化方法仍然有限。本研究提出了一种实验室间比较(ILC),旨在评估使用微拉曼光谱和具有代表性的聚对苯二甲酸乙二醇酯(PET)标准物质(RM)鉴定和定量婴儿奶粉中小MPs (5-100 μ m)的分析方法的准确性和可比性。制剂为水溶性片剂,旨在复制环境相关MPs的形态、大小分布和聚合物组成,并预先评估了质量分数和颗粒数的均匀性和稳定性。采用两个具有不同MPs颗粒数的PET RM批次(高负载批次:1759±141 MPs;低负载批次:160±22 MPs)进行酶化学消化,然后在两个实验室使用不同的仪器和操作人员独立进行µ拉曼分析。结果报告为每个分析样品的绝对颗粒计数,并且在所有尺寸类别中都表现出出色的回收率,包括最小的颗粒(低至5 μ m),回收率从82%到88%,与RM参考值很好地一致。该分析方法被证明是稳健的、可重复的,并且适用于复杂食品基质中的低水平多磺酸盐定量,为食品部门可靠的多磺酸盐监测方法的统一和标准化提供了支持。
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引用次数: 0
Validation of combination of protodioscin and shatavarin iv from medicinal extracts for alleviating menopausal symptoms by computational deep learning models 通过计算深度学习模型验证药用提取物中原薯蓣皂苷和沙伐林iv联合用于缓解更年期症状的有效性
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-09-08 DOI: 10.1016/j.talo.2025.100552
Rithika Naveencharan , Ashwin Sivakumar , Rishi Senthil Kumar , Sheena Christabel Pravin , Reena Monica P , V. Kiruthika , Abhijeet Morde , Arun Balakrishnan , Muralidhara Padigaru , Ninad Puranik , Ravindra Nayakwadi , Tejas Namjoshi
Shatavarin-IV and Protodioscin, saponins of Asparagus racemosus and Trigonella foenum-graecum respectively are phytoestrogens commonly used to alleviate post-menopausal symptoms such as hot flashes, sleeping disorder, and mood sways. While their individual benefits are well-documented, the synergistic effects of these compounds remain underexplored. This study aims to validate the combination of Shatavarin-IV and Protodioscin on molecular mediators of menopausal symptoms using advanced computational analyses. Deep learning model for drug additivity analysis was developed and trained on the DrugComb database using the Loewe’s additivity score as the target variable, with a focus on combinations tested on the MCF7 cell line and validated of the 786-O cell line. Molecular docking simulations were performed using PyRx, PyMOL, FPocket and ChimeraX to investigate the binding affinities of Protodioscin and Shatavarin-IV with key neuronal modulators of hot flashes, including Kisspeptin, Neurokinin B, TRPV1, c-FOS, and GnRH. IC50 values were derived from binding free energies. Dose-response simulations were evaluated across concentrations (0.01 µM to 100 µM) using a four-parameter logistic equation. Absorption, Distribution, Metabolism, and Elimination (ADME) profiling was determined to predict bioavailability and toxicity. The dose-response simulations confirmed significant therapeutic potential at relevant concentrations and additive interaction between both the compounds suggests enhanced therapeutic efficacy. ADME profiling indicated favorable bioavailability, non-toxicity, and efficient metabolism for both compounds and provides novel insights into their molecular mechanism of action in the body. The results of this research indicate that the combined additive effect of Protodioscin and Shatavarin-IV in alleviating menopausal symptoms exhibits favorable pharmacokinetic profile, and could potentially be promising candidates for future human clinical studies.
总状芦笋皂苷和三角芦笋皂苷原薯蓣皂苷分别是植物雌激素,常用于缓解绝经后潮热、睡眠障碍、情绪波动等症状。虽然它们各自的益处已被充分证明,但这些化合物的协同效应仍未得到充分探讨。本研究旨在通过先进的计算分析验证Shatavarin-IV和原薯蓣皂苷对绝经期症状分子介质的联合作用。以Loewe加性评分为目标变量,在DrugComb数据库上开发并训练了药物加性分析的深度学习模型,重点对MCF7细胞系和786-O细胞系进行了组合测试和验证。利用PyRx、PyMOL、FPocket和ChimeraX进行分子对接模拟,研究Protodioscin和Shatavarin-IV与热潮热的关键神经元调节剂Kisspeptin、Neurokinin B、TRPV1、c-FOS和GnRH的结合亲和力。IC50值由束缚自由能导出。使用四参数logistic方程评估不同浓度(0.01µM至100µM)的剂量-反应模拟。吸收、分布、代谢和消除(ADME)分析被确定预测生物利用度和毒性。剂量-反应模拟证实了在相关浓度下显著的治疗潜力,两种化合物之间的加性相互作用表明治疗效果增强。ADME分析表明,这两种化合物具有良好的生物利用度、无毒性和有效的代谢,并为其在体内的分子作用机制提供了新的见解。本研究结果表明,原diooscin和Shatavarin-IV在缓解绝经期症状方面的联合加性作用表现出良好的药代动力学特征,可能是未来人类临床研究的潜在有希望的候选者。
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引用次数: 0
SensEDNA: An innovative optical CRISPR platform for rapid environmental DNA monitoring SensEDNA:一个创新的光学CRISPR平台,用于快速环境DNA监测
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-09-10 DOI: 10.1016/j.talo.2025.100551
Molly Ann Williams , Ciprian Briciu-Burghina , Sean Power , Joyce O’Grady , Elvira de Eyto , Nigel Kent , Anne Parle-McDermott , Fiona Regan
Biodiversity loss is at an all-time high increasing the need for simple and rapid species monitoring systems. Management and conservation of fish species within aquatic environments requires knowledge of distribution, traditionally gained through visual detection and counting. These methods are expensive, time consuming and can lead to harm of the species of interest. Environmental DNA (eDNA) offers a solution to this through detection of DNA that may be shed into the environment by a given target species. Integrating eDNA based molecular assays to a biosensor device enables onsite sample testing for rapid species assessment.
Herein, a portable fluorometer with incubating capabilities (SensEDNA) was custom designed and built to conduct the simultaneous incubation and fluorescent detection steps required for isothermal eDNA detection. The system features three optical cells to enable triplicate analysis. Each optical cell is set to incubate at 37 °C and control within 0.5 °C. Fluorescence detection with excitation at 485 nm and emission at 535 nm was integrated and optimised to provide highest signal/noise ratio while minimising fluorescence bleaching. The system comprises a simple LED/high pass filter/photodiode configuration and is controlled by a Wixel board. This affordable and user-friendly optical detection platform, coupled with RPA-CRISPR-Cas, provides a crucial step towards on-site single species identification from eDNA.
生物多样性的丧失正处于空前的高度,增加了对简单和快速物种监测系统的需求。水生环境中鱼类的管理和保护需要有关分布的知识,传统上是通过目测和计数获得的。这些方法成本高,耗时长,而且可能导致对感兴趣的物种的伤害。环境DNA (eDNA)通过检测可能由给定目标物种释放到环境中的DNA提供了解决方案。将基于eDNA的分子分析集成到生物传感器设备中,可以进行现场样品测试,以进行快速物种评估。为此,我们定制了一种具有孵育功能的便携式荧光仪(SensEDNA),用于同时进行等温eDNA检测所需的孵育和荧光检测步骤。该系统具有三个光学单元,可以进行三次重复分析。每个光学细胞设置在37°C孵育,控制在0.5°C内。集成并优化了485 nm激发和535 nm发射的荧光检测,以提供最高的信噪比,同时最大限度地减少荧光漂白。该系统包括一个简单的LED/高通滤波器/光电二极管配置,并由一个Wixel板控制。这种价格合理且用户友好的光学检测平台,加上RPA-CRISPR-Cas,为eDNA的现场单物种鉴定提供了关键的一步。
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引用次数: 0
Design and fabrication of microfluidic chip and integrated nucleic acid rapid diagnosis device 微流控芯片及集成核酸快速诊断装置的设计与制造
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-10-24 DOI: 10.1016/j.talo.2025.100582
Wenju Ren , Chenyang Qi , Rui Yang , Jie Zhou , Liu Yang , Yuandi Yu , Taixiong Zheng ([email protected]) , Lizhi Fu
African Swine Fever (ASF) poses a severe threat to the global pig husbandry. In the absence of an effective vaccine, developing rapid and accurate diagnostic methods is crucial for epidemic control. This study designed a low-cost microfluidic chip and integrated nucleic acid rapid detection device (INARDD) based on colorimetric principles combined with magnetic bead based nucleic acid extraction technology. Manufactured using micro computer numerical control machine tools (CNC) and 3D printing, the device fully automates the entire process include nucleic acid extraction, transfer, loop-mediated isothermal amplification (LAMP), and detection without requiring external equipment or manual intervention. Experimental results demonstrate high sensitivity (94.1 %, 32/34), specificity (97.7 %, 42/43), and accuracy (96.1 %, 74/77), with a detection limit of 10² copies/μL. The device features low production costs (approximately $69.1), low per-test expenses, and a portable design (2 kg), facilitating field deployment. This study provides an economical and efficient solution for rapid on-site diagnosis of African swine fever, holding significant implications for enhancing epidemic control and ensuring meat safety. It also offers technical reference for developing portable diagnostic systems for other infectious diseases.
非洲猪瘟(ASF)对全球养猪业构成严重威胁。在缺乏有效疫苗的情况下,开发快速和准确的诊断方法对于流行病控制至关重要。本研究设计了一种基于比色法原理结合磁珠核酸提取技术的低成本微流控芯片及集成核酸快速检测装置(INARDD)。该设备采用微型计算机数控机床(CNC)和3D打印制造,完全自动化了整个过程,包括核酸提取、转移、环介导等温扩增(LAMP)和检测,无需外部设备或人工干预。实验结果表明,该方法灵敏度为94.1%(32/34),特异度为97.7%(42/43),准确度为96.1%(74/77),检出限为10²copies/μL。该设备的特点是生产成本低(约69.1美元),每次测试费用低,便携式设计(2公斤),便于现场部署。本研究为非洲猪瘟现场快速诊断提供了一种经济高效的解决方案,对加强疫情控制和确保肉类安全具有重要意义。为开发其他传染病便携式诊断系统提供了技术参考。
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引用次数: 0
Development of molecularly imprinted polymers via dual polymerisation strategies for targeted isolation of Ethyl p-Methoxycinnamate from Kaempferia galanga L. extract 双聚合技术在山柰提取物中对甲氧基肉桂酸乙酯分子印迹聚合物中的应用
IF 3.7 Q1 CHEMISTRY, ANALYTICAL Pub Date : 2025-12-01 Epub Date: 2025-10-18 DOI: 10.1016/j.talo.2025.100580
Marisa Dwi Ariani , Ade Zuhrotun , Panagiotis Manesiotis , Aliya Nur Hasanah
The rhizome of Kaempferia galanga L. contained ethyl p-methoxycinnamate (EPMC) as a major component of its essential oil. Despite the abundance of EPMC in the plant, conventional isolation methods yielded only 0.5 – 2.5 %. This study developed molecularly imprinted polymers (MIPs) via bulk and suspension polymerisation to enhance the selective isolation of EPMC from K. galanga extracts. Six functional monomers were screened for their binding affinity with EPMC. 2-hydroxyethyl methacrylate (HEMA) in chloroform and methacrylic acid (MAA) in n-hexane were selected for further investigation. Stoichiometric analysis established optimal template-to-monomer ratios of 1:6 for HEMA and 1:7 for MAA. Eight MIP formulations and their corresponding non-imprinted polymers (NIPs), were synthesised using these monomers via both polymerisation methods. Characterisation using Fourie Transform Infra-Red (FTIR), Scanning Electron Microscope (SEM), Brunauer-Emmett-Teller (BET), and Particle Size Analysis (PSA) revealed that bulk polymers exhibited larger, irregular, and non-uniform particles compared to those produced by suspension polymerisation. Adsorption studies confirmed that the MIPs follow Freundlich isotherms, with MIP B2 (bulk, MAA, 1:7 ratio) exhibiting the highest binding affinity (KF = 0.081 mg/g). MIP B2 also demonstrated superior performance in the solid-phase extraction of EPMC from extracts, achieving recoveries of up to 82.4 % ± 5.52 and imprinting factors above 1.3. Selectivity tests confirmed strong discrimination of EPMC over structural analogues. In conclusion, MIP B2 offers a selective, efficient, and scalable method for EPMC isolation. These findings supported the continued development of tailored MIPs for natural product purification and provided a foundation for future optimisation of monomer-initiator systems and polymerisation parameters.
山柰根茎精油的主要成分是对甲氧基肉桂酸乙酯(EPMC)。尽管EPMC在植物中含量丰富,但传统的分离方法只能得到0.5 - 2.5%。为了提高高良姜提取物中EPMC的选择性分离,研究了分子印迹聚合物(MIPs)的制备方法。筛选了6个功能单体与EPMC的结合亲和力。以氯仿中的2-甲基丙烯酸羟乙酯(HEMA)和正己烷中的甲基丙烯酸(MAA)为研究对象。化学计量学分析确定HEMA和MAA的最佳模板-单体比分别为1:6和1:7。利用这些单体通过两种聚合方法合成了8种MIP配方及其相应的非印迹聚合物(NIPs)。利用傅里变换红外(FTIR)、扫描电镜(SEM)、布鲁诺尔-埃米特-泰勒(BET)和粒度分析(PSA)进行表征表明,与悬浮聚合产生的聚合物相比,大块聚合物表现出更大、不规则和不均匀的颗粒。吸附研究证实,MIP符合Freundlich等温线,其中MIP B2(体积,MAA, 1:7比)具有最高的结合亲和力(KF = 0.081 mg/g)。MIP B2固相萃取EPMC的回收率高达82.4%±5.52,印迹因子在1.3以上。选择性试验证实了EPMC对结构类似物的强烈区分。总之,MIP B2提供了一种选择性、高效和可扩展的EPMC分离方法。这些发现支持了用于天然产物纯化的定制mip的持续发展,并为未来优化单体引发剂系统和聚合参数提供了基础。
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引用次数: 0
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Talanta Open
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