Name: a simple method for automated solid phase extraction of water samples for immunological analysis of small pollutants
Uploaded: 2016-01-01T14:00:00.000+00:00
Duration: 7 min 26 s
Description: Watch this Scientific Journal Video about A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants at JoVE.com

This work was funded by the European Union Seventh Framework Program FP7/2007-2013 under grant agreement no. 265721. The authors thank the RIKILT Institute for Food Safety (NL) for their support in this project.

注意:下面的协议说明了SPE 100毫升的水样品与C18的吸附剂和洗脱用50％体积/体积甲醇进行。富集的样品稀释分析用酶联免疫吸附试验（ELISA）试剂盒的酶之前达到10％体积/体积甲醇。 1.准备试剂<ol><li>制备水样<ol><li>之前的任何其它步骤，筛选各100毫升的水样品与0.2μm孔径的过滤器。 </li><li>通过稀释的参考溶液适当体积入水体积穗与期望浓度的样品。例如，制备100毫升的水样品与E2的100纳克/升的稀释3.3微升的E2参考溶液在300微克/升的浓度。稀释此溶液十次以获得一个样品掺入10纳克/升的E2。稀释后者另一个十倍，得到100ml的样品与1纳克/升的E2。 </li><li>放置过滤后的样品（未修饰的或加标）在玻璃瓶中以GL45线程。使用示例在同一天。取一小部分，以与ELISA法进行分析，来估计初始浓度。 </li></ol></li><li>通过在去离子（二 - ）水在紧管稀释甲醇至50％体积/体积制备300μl的洗脱液。 </li></ol> 2.制备固相萃取柱的<ol><li>通过加入第一1600微升甲醇中，然后加入400μl的二 - 水至40mg反相吸附剂的制备20毫克/毫升悬浮十八烷基二氧化硅吸附剂颗粒。盖紧盖子，搅拌用旋涡。 </li><li>安装在塔底部的膜<ol><li>选择一个尼龙膜孔径为11微米，将其放置在防尘组织构成的双层。用3毫米直径冲头，切断在膜2的小零件。 </li><li>抓住小膜用平头镊子过滤器之一，并将其​​放置在列的一侧。 </li><li>拧平底接头与管并拧紧。该膜为n流到位和吸着能够可靠地加入。上朝向的最后一列指示的体绘制的箭头，其中膜置于。 </li></ol></li><li>准备填充吸附柱<ol><li>固定柱向底部的箭头指向在支架上。列必须是垂直地设置。 </li><li>附加空10毫升一次性注射器到列的管的端部，通过使用鲁尔锁定连接​​器。 </li><li>准备微量移液器用20-200微升小费，定容至100微升。此枪头格式适于吸附剂柱的要制备的大小。该程序将具有较大的枪头，如用1毫升吸管中使用的那些更加困难。 </li><li>搅动吸附剂悬浮液用涡旋，并迅速吸管100微升在中心上的列。同时注入，吸轻轻所有移液溶液通过使用注射器用另一只手槽膜。在这届年龄，溶液灌装注射器必须明确微粒，粒子床最终可以在列中观察到。 </li><li>通过搅拌所有移液步骤之间的纸浆悬浮液，以确保在溶液中的颗粒的均匀悬浮液重复此过程2次。由此产生的列包含吸附剂6毫克。 </li><li>当将300μl悬浮液已被加载并通过用注射器抽吸干燥，保持注射器在适当位置，并放置在顶部的第二尼龙膜通过使用钳子用另一只手。 </li><li>拧紧第二个连接器管和处置注射器。该固相萃取柱就可以使用了。 </li></ol></li></ol> 3.准备系统<ol><li>通过使用鲁尔锁接头拧紧设备上的固相萃取柱。箭头必须是指向相同的方向在设备上显示的。 </li><li>通过拧紧连接含有样品到装置中的瓶它提供了GL45安全帽。 </li><li>加载200微升洗脱液中的"淋洗液"水库。 </li><li>负载800微升二水的"稀释"水库。 </li><li>将一个瓶在排污口在提取步骤收集处理过的水。格式并不重要，但体积需要足够大的方面的样品体积。 </li><li>将小瓶在1.5毫升体积最小容量的传感器出口。 1 mL的最大容积将过小，收集丰富洗脱液和稀释液时气泡会形成在该出口。 </li><li>验证压力调节器处于关闭位置时通过手动转动它，反向旋转，直至进一步的移动是不可能的。 </li></ol> 4. SPE与原型<ol><li>通过按下背面的按钮开关上的原型。 </li><li>制备用户界面和选择程序<ol><li>打开用户界面。选择计算机的通讯端口来进行设备从列表连接，然后点击按钮"下一步"。 </li><li>输入值580 PSET和30 dpset。泵将调整运行时，以维持在加压系统和储层压力，在580±30毫巴。 </li><li>选择自动模式。 </li><li>在自动模式下的角落，装入箱子"配置文件路径"程序文件。 </li></ol></li><li>调节压力调节器<ol><li>启动泵。 </li><li>手动转动压力调节器，直到读取预浸料值小于但接近320毫巴。 </li><li>停止泵。 </li></ol></li><li>启动SPE程序<ol><li>按在自动模式下的角落"开始"。泵将接通和提取，洗脱和稀释步骤将自动跟随另一个。在50分钟中进行，整个过程在100毫升样品。 </li><li>验证预浸的值。它必须是在范围320 - 350毫巴在提取步骤以确保最佳的流速。 </li><li>关闭小瓶并储存在4-5°C黑暗直至分析。在下面的30小时进行分析，以防止分析物的降解。 </li><li>处置处理过的水。 </li></ol></li><li>清洁系统 注意:在待清洁每次萃取步骤的系统需要防止交叉污染。 <ol><li>制备甲醇70％体积/体积的在一个GL 45玻璃瓶10ml的溶液中。 </li><li>拔下固相萃取柱和插头与连接器的管道。 </li><li>在自动模式下，选择"清洗"文件和相同的压力设置启动。 </li></ol></li></ol> 5.检测与ELISA雌二醇浓度<ol><li>制备校准样品浓度在设有所使用的ELISA试剂盒的协议表示。准备一张CALIBRA中置具有相同的矩阵作为样品，和一个校准设置用甲醇10％体积/体积。 </li><li>分配在盘上的孔中的样品的所需量，根据制造商的说明。使用校准样品，过滤但未处理与SPE，和丰富的样品甲醇10％V不变并加标水样/ V。使用每样本3个孔，以减少与测定相关的误差。 </li><li>遵循即随试剂盒提供的反应物的加入，培养时间，洗涤和进一步反应与酶的协议的指示。 </li><li>读出的光信号在每个孔中根据试剂盒制造商的用酶标仪仪器指令和收集数据。 </li><li>使用平板读取器仪器的软件，以适合校准曲线，并确定与相同矩阵的初始样品中的E2浓度，并且在富集样品用甲醇10％体积/体积。 </li></ol>

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The authors have nothing to disclose.

提出了水样然后分析使用免疫的制备新方法。该仪器能够以自动化和用户友好的方式进行固相萃取。 水样其注射之前到系统的过滤是非常关键的。溶液中存在的任何微粒仍然会潜在地引起流体网络的堵塞和阻碍SPE柱。另一个重要步骤是在SPE柱的制备。在列颗粒的量是至关重要的，以达到最佳的性能成为可能。制备吸附剂颗粒的悬浮液时，避免了粒子的附聚特必须小心​​。?...

样品制备是在任何分析过程中的重要一步。特别是，除去基体效应，干扰的缩减，且分析物的富集是必要的，以获得精确的结果，并达到检测的低限。内分泌干​​扰化合物（内分泌干扰物）是特别值得关注的，因为它们对活的生物体的动作，即使在环境中存在的非常低的水平。天然激素17β雌二醇存在于欧盟水污染的收藏和容易被加入到根据欧洲水框架指令调节优先物质清单。固相萃取（SPE）通常适用于小的污染物在水中的分析，与两个化学1-5（色谱，质谱）和免疫6-9检测方法。后者积累了环境监测领域的兴趣，如免疫测定法是在大量的各种格式提供，特定于たrget分析物，和达到低检测极限。6，7，10，11的各种酶联免疫吸附测定（ELISA）是可商购，并使得能够分析多个样品同时上的多孔板。该过程包括在连续的反应步骤，可能需要几个小时。反应的最终产物，可以检测光学来确定基于校准曲线对目标分子的浓度。 经典的SPE程序包括吸附剂预调节，样品萃取，清洗，洗脱，浓缩由洗脱液蒸发。用于稀释该提取物的溶剂选自根据检测方法。对于免疫学方法中，有机溶剂的影响强烈的方法的灵敏度的量。12 除了回收和预浓缩的表演中，该方法也需要简单和成本效益。在再修改的自动化Ë有助于减少人类相关的错误。在我们以前的工作13我们推出了样机的自动固相萃取，而我们的方法适用于天然激素17β雌二醇海水样品中的分析。在本视频中，我们想强调我们方法的技术优势相对于传统的离线和在线SPE，以及它与通过检测免疫反应的具体的兼容性。我们描述适用于水样品17β雌二醇的检测的协议。 SPE与十八烷基硅胶（C18）吸附剂相和洗脱用稀甲醇进行执行。

<table><tbody><tr><td>Filter membrane 0.2 &amp;mu;m pore size</td><td>Merck Millipore</td><td>GNWP04700</td><td>For sample filtration</td></tr><tr><td>Nylon membrane 11 &amp;mu;m pore size</td><td>Merck Millipore</td><td>NY1104700</td><td>For SPE column</td></tr><tr><td>Disposable biopsy punch 5 mm</td><td>Medical Budget</td><td>39302439</td><td></td></tr><tr><td>Nucleodur C18 ec&amp;nbsp;</td><td>Macherey Nagel</td><td>713550.01</td><td>50 &amp;mu;m particle diameter</td></tr><tr><td>Synthetic sea water</td><td>Sigma Aldrich</td><td>SSWS500-500ML</td><td></td></tr><tr><td>Methanol</td><td>VWR</td><td></td><td></td></tr><tr><td>17beta-estradiol standard</td><td>Enzo Life Science</td><td></td><td>300 ng/ml</td></tr><tr><td>17beta-estradiol ELISA kit</td><td>Enzo Life Science</td><td>ADI-900-008</td><td>96 wells, range 30 - 3,000 ng/L</td></tr></tbody></table>

a simple method for automated solid phase extraction of water samples for immunological analysis of small pollutants

对环境水样的固相萃取（SPE）的新方法。所开发的原型是成本效益和用户友好的，并能进行快速，自动化和简单的SPE。预浓缩的溶液与通过免疫测定分析兼容，具有低的有机溶剂含量。一种方法被描述为天然激素17β雌二醇在100ml水样提取和预浓缩。反相SPE执行与十八烷基二氧化硅吸附剂和洗脱用200μl甲醇50％体积/体积的完成。洗脱剂是通过将二 - 水，以降低甲醇的量稀释。手动制备SPE柱后，整体过程1小时内自动执行。在该过程结束时，雌二醇浓度通过使用市售酶联免疫吸附测定（ELISA）进行测定。 100倍预浓缩得以实现，在只有10％体积/体积的甲醇含量。完全恢复的分子用1ng得以实现/升掺入去离子和合成海水样品。

吸附剂填料的再现性通过干燥评价和加权在玻璃小瓶的移液吸附剂，结果示于图1。重复性的喷射时100ml中的样品进行了测试，如在初始和预如图 2所示。浓度浓缩加标样品通过使用商业ELISA试剂盒17β雌二醇测定，并示于图3。 所提出的方法包括在制备吸附剂相（ <st...

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A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

对水样的自动固相萃取小污染物的免疫学分析的简单方法

A new method for solid phase extraction (SPE) of environmental water samples is proposed. The developed prototype is cost-efficient and user friendly, and ...

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13.7K 次观看.  Centre Suisse d'Electronique et de Microtechnique. A protocol for the extraction and pre-concentration of estradiol from water samples by using an automated and miniaturized system is presented.

视频: A Simple Method for Automated Solid Phase Extraction of Water Samples for Immunological Analysis of Small Pollutants

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA libraries are needed for optimal results from next-generation sequencing. Environmental samples such as water may have low quality and quantities of DNA as well as contaminants that co-precipitate with DNA. The mechanical and enzymatic processes involved in extraction and library preparation may further damage the DNA. Gel size selection enables purification and recovery of DNA fragments of a defined size for sequencing applications. Nevertheless, this task is one of the most time-consuming steps in the DNA library preparation workflow. The protocol described here enables complete automation of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments. 
In this study, we describe a high-throughput approach to prepare high quality DNA libraries from freshwater samples that can be applied also to other environmental samples. We used an indirect approach to concentrate bacterial cells from environmental freshwater samples; DNA was extracted using a commercially available DNA extraction kit, and DNA libraries were prepared using a commercial transposon-based protocol. DNA fragments of 500 to 800 bp were gel size selected using Ranger Technology, an automated electrophoresis workstation. Sequencing of the size-selected DNA libraries demonstrated significant improvements to read length and quality of the sequencing reads.

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

自动凝胶尺寸选择，以提高下一代测序文库的质量，从环境水样准备

Next-generation sequencing of environmental samples can be challenging because of the variable DNA quantity and quality in these samples. High quality DNA ...

科研

环境科学

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

准备和测试阻抗为基础与RTgill-W1细胞饮用水进行快速评估水样进行毒性射流生物芯片

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

Characterization and Application of Passive Samplers for Monitoring of Pesticides in Water

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study provides a protocol for the passive sampler preparation, calibration, extraction method and instrumental analysis. Sampling rates (RS) and passive sampler-water partition coefficients (KPW) were calculated for silicone rubber, polar organic chemical integrative sampler POCIS-A, POCIS-B, SDB-RPS and C18 disk. The uptake of the selected compounds depended on their physicochemical properties, i.e., silicone rubber showed a better uptake for more hydrophobic compounds (log octanol-water partition coefficient (KOW) &#62; 5.3), whereas POCIS-A, POCIS-B and SDB-RPS disk were more suitable for hydrophilic compounds (log KOW &#60; 0.70).

A protocol about the characterization and application of five different passive sampling devices is presented.

表征和被动采样的应用在农药水监测

Five different water passive samplers were calibrated under laboratory conditions for measurement of 124 legacy and current used pesticides. This study ...

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and detritus, and excreting particulate and dissolved compounds, they serve as important agents for benthic-pelagic coupling. Accurately measuring the compounds removed and excreted by suspension feeders (such as sponges, ascidians, polychaetes, bivalves) is crucial for the study of their physiology, metabolism, and feeding ecology, and is fundamental to determine the ecological relevance of the nutrient fluxes mediated by these organisms. However, the assessment of the rate by which suspension feeders process particulate and dissolved compounds in nature is restricted by the limitations of the currently available methodologies. Our goal was to develop a simple, reliable, and non-intrusive method that would allow clean and controlled water sampling from a specific point, such as the excurrent aperture of benthic suspension feeders, in situ. Our method allows simultaneous sampling of inhaled and exhaled water of the studied organism by using minute tubes installed on a custom-built manipulator device and carefully positioned inside the exhalant orifice of the sampled organism. Piercing a septum on the collecting vessel with a syringe needle attached to the distal end of each tube allows the external pressure to slowly force the sampled water into the vessel through the sampling tube. The slow and controlled sampling rate allows integrating the inherent patchiness in the water while ensuring contamination free sampling. We provide recommendations for the most suitable filtering devices, collection vessel, and storing procedures for the analyses of different particulate and dissolved compounds. The VacuSIP system offers a reliable method for the quantification of undisturbed suspension feeder metabolism in natural conditions that is cheap and easy to learn and apply to assess the physiology and functional role of filter feeders in different ecosystems.

VacuSIP, an Improved InEx Method for In Situ Measurement of Particulate and Dissolved Compounds Processed by Active Suspension Feeders

We introduce the VacuSIP, a simple, non-intrusive, and reliable method for clean and accurate point sampling of water. The system was developed and evaluated for the simultaneous collection of the water inhaled and exhaled by benthic suspension feeders in situ, to cleanly measure removal and excretion of particulate and dissolved compounds.

VacuSIP，对改进的方法INEX<em&gt;原位</em&gt;颗粒物与测量溶解的化合物加工后主动悬挂给料机

Benthic suspension feeders play essential roles in the functioning of marine ecosystems. By filtering large volumes of water, removing plankton and ...

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better characterize the prevalence of these pathogens, EPA Method 1623 was developed and used to monitor levels of these organisms in US drinking water supplies 12. The method has three main parts; the first is the sample concentration in which at least 10 L of raw surface water is filtered. The organisms and trapped debris are then eluted from the filter and centrifuged to further concentrate the sample. The second part of the method uses an immunomagnetic separation procedure where the concentrated water sample is applied to immunomagnetic beads that specifically bind to the Cryptosporidium oocysts and Giardia cysts allowing for specific removal of the parasites from the concentrated debris. These (oo)cysts are then detached from the magnetic beads by an acid dissociation procedure. The final part of the method is the immunofluorescence staining and enumeration where (oo)cysts are applied to a slide, stained, and enumerated by microscopy.
Method 1623 has four listed sample concentration systems to capture Cryptosporidium oocysts and Giardia cysts in water: Envirochek filters (Pall Corporation, Ann Arbor, MI), Envirochek HV filters (Pall Corporation), Filta-Max filters (IDEXX, Westbrook, MA), or Continuous Flow Centrifugation (Haemonetics, Braintree, MA). However, Cryptosporidium and Giardia (oo)cyst recoveries have varied greatly depending on the source water matrix and filters used1,14. A new tangential flow hollow-fiber ultrafiltration (HFUF) system has recently been shown to be more efficient and more robust at recovering Cryptosporidium oocysts and Giardia cysts from various water matrices; moreover, it is less expensive than other capsule filter options and can concentrate multiple pathogens simultaneously1-3,5-8,10,11. In addition, previous studies by Hill and colleagues demonstrated that the HFUF significantly improved Cryptosporidium oocysts recoveries when directly compared with the Envirochek HV filters4. Additional modifications to the current methods have also been reported to improve method performance. Replacing the acid dissociation procedure with heat dissociation was shown to be more effective at separating Cryptosporidium from the magnetic beads in some matrices9,13 .
This protocol describes a modified Method 1623 that uses the new HFUF filtration system with the heat dissociation step. The use of HFUF with this modified Method is a less expensive alternative to current EPA Method 1623 filtration options and provides more flexibility by allowing the concentration of multiple organisms.

A Modified EPA Method 1623 that Uses Tangential Flow Hollow-fiber Ultrafiltration and Heat Dissociation Steps to Detect Waterborne Cryptosporidium and Giardia spp.

This protocol describes the use of a tangential flow hollow-fiber ultrafiltration sample concentration system and a heat dissociation as alternative steps for the detection of waterborne Cryptosporidium and Giardia species using EPA Method 1623.

一种改进的EPA方法1623使用切向流中空纤维超滤和热解离步骤检测水性<em&gt;隐</em&gt;和<em&gt;贾第虫属。</em

Cryptosporidium and Giardia species are two of the most prevalent protozoa that cause waterborne diarrheal disease outbreaks worldwide. To better ...

免疫与感染

Identifying Per- and Polyfluorinated Chemical Species with a Combined Targeted and Non-Targeted-Screening High-Resolution Mass Spectrometry Workflow

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the local to federal levels. The continuing evolution of PFAS chemistries presents a challenge to the environmental monitoring, where ongoing development of targeted methods necessarily lags the discovery of new chemical compounds. There is a need, therefore, to have forward-looking methodologies that can detect emerging and unexpected compounds, monitor these species over time, and resolve details of their chemical structure to enable future work in human health. To this end, non-targeted analysis by high-resolution mass spectrometry offers a broad base detection approach that can be combined with almost any sample preparation scheme and provides significant capabilities for compound identification after detection. Herein, we describe a solid-phase extraction (SPE) based sample concentration method tuned for shorter chain and more hydrophilic PFAS chemistries, such as per fluorinated ether acids and sulfonates, and describe analysis of samples prepared in this fashion in both targeted and non-targeted modes. Targeted methods provide superior quantification when reference standards are available but are intrinsically limited to expected compounds when performing analysis. In contrast, a non-targeted approach can identify the presence of unexpected compounds and provide some information about their chemical structure. Information about chemical features can be used to correlate compounds across sample locations and track abundance and occurrence over time.

Here, we present a protocol for the sequential targeted quantification and non-targeted analysis of fluorinated compounds in water by mass spectrometry. This methodology provides quantitative levels of known fluorochemical compounds and identifies unknown chemicals in related samples with semi-quantitative estimates of their abundance.

利用目标筛选和非靶向筛选高分辨率质谱相结合的工作流程识别跨氟和多氟化学物种

Historical and emerging per- and polyfluoroalkyl substances (PFASs) have garnered significant interest from the public and government agencies from the ...

Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water samples. A brief overview of the evolution of chip-based SPE techniques is provided. This is followed by an introduction to specific polymeric materials and their role in SPE. To develop an innovative dipole-assisted SPE technique, a chlorine (Cl)-containing SPE functionality was implanted into a poly(methyl methacrylate) (PMMA) microchip. Herein, diverse analytical techniques including contact angle analysis, Raman spectroscopic analysis, and laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) analysis were employed to validate the utility of the implantation protocol of the C-Cl moieties on the PMMA. The analytical results of the X-ray absorption near-edge structure (XANES) analysis also demonstrated the feasibility of the Cl-containing PMMA used as an extraction medium by virtue of the dipole-ion interactions between the highly electronegative C-Cl moieties and the positively charged metal ions.

The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

水样中偶极辅助固相萃取Microchip的痕量金属分析的制作

This paper describes a fabrication protocol for a dipole-assisted solid phase extraction (SPE) microchip available for trace metal analysis in water ...

生物工程

Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay

Electrowetting is the effect by which the contact angle of a droplet exposed to a surface charge is modified. Electrowetting-on-dielectric (EWOD) exploits the dielectric properties of thin insulator films to enhance the charge density and hence boost the electrowetting effect. The presence of charges results in an electrically induced spreading of the droplet which permits purposeful manipulation across a hydrophobic surface. Here, we demonstrate EWOD-based protocol for sample processing and detection of four categories of antigens, using an automated surface actuation platform, via two variations of an Enzyme-Linked Immunosorbent Assay (ELISA) methods. The ELISA is performed on magnetic beads with immobilized primary antibodies which can be selected to target a specific antigen. An antibody conjugated to HRP binds to the antigen and is mixed with H2O2/Luminol for quantification of the captured pathogens. Assay completion times of between 6 and 10 min were achieved, whilst minuscule volumes of reagents were utilized.

Electrowetting-based digital microfluidic is a technique that utilizes a voltage-driven change in the apparent contact angle of a microliter-volume droplet to facilitate its manipulation. Combining this with functionalized magnetic beads enables the integration of multiple laboratory unit operations for sample preparation and identification of pathogens using Enzyme-linked Immunosorbent Assay (ELISA).

基于电润的数字微流体平台，用于自动酶链接免疫吸附测定

Electrowetting is the effect by which the contact angle of a droplet exposed to a surface charge is modified. Electrowetting-on-dielectric (EWOD) exploits ...

Solid-Liquid Extraction

Source: Laboratory of Dr. Jay Deiner &#8212; City University of New York
Extraction is a crucial step in most chemical analyses. It entails removing the analyte from its sample matrix and passing it into the phase required for spectroscopic or chromatographic identification and quantification. When the sample is a solid and the required phase for analysis is a liquid, the process is called solid-liquid extraction. A simple and broadly applicable form of solid-liquid extraction entails combining the solid with a solvent in which the analyte is soluble. Through agitation, the analyte partitions into the liquid phase, which may then be separated from the solid through filtration. The choice of solvent must be made based on the solubility of the target analyte, and on the balance of cost, safety, and environmental concerns.

Solid-Liquid Extraction: Principle, Procedure, Applications

固-液萃取

Source: Laboratory of Dr. Jay Deiner &#8212; City University of New York
Extraction is a crucial step in most chemical analyses. It entails removing the ...

教育

JoVE Science Education

化学

有机化学

Extraction of Biomarkers from Sediments - Accelerated Solvent Extraction

Source: Laboratory of Jeff Salacup&#160;- University of Massachusetts Amherst
The distribution of a group of organic biomarkers called glycerol-dialkyl glycerol-tetraethers (GDGTs), produced by a suite of archaea and bacteria, were found in modern sediments to change in a predictable manner in response to air or water temperature1,2. Therefore, the distribution of these biomarkers in a sequence of sediments of known age can be used to reconstruct the evolution of air and/or water temperature on decadal to millennial timescales (Figure 1). The production of long high-resolution records of past climates, called paleoclimatology, depends on the rapid analysis of hundreds, possibly thousands of samples. Older extraction techniques, such as sonication or Soxhlet, are too slow. However, the newer Accelerated Solvent Extraction technique was designed with efficiency in mind.
<img alt="Figure 1" src="/files/ftp_upload/10097/10097fig1.jpg" /> 
Figure 1. An example of a paleoclimate record showing changes in sea surface temperature (SST) in the eastern Mediterranean Sea during the past ~27,000 years3. This record comprises ~115 samples and is based on the isoprenoidal GDGT-based TEX86 SST proxy.

Accelerated Solvent Extraction of Total Lipid from Sediments

生物标志物提取沉积物-加速溶剂萃取

Source: Laboratory of Jeff Salacup&#160;- University of Massachusetts Amherst
The distribution of a group of organic biomarkers called glycerol-dialkyl ...

对水样的自动固相萃取小污染物的免疫学分析的简单方法

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