The authors would like to convey their gratitude for the technical support provided by National Synchrotron Radiation Research Center (NSRRC) (Taiwan). The authors are grateful for the financial support provided by the Ministry of Science and Technology of the Republic of China (Taiwan) and the Industrial Technology Research Institute (Taiwan).

注意！一些化学物质（ 如丙烯酰胺，1,1&#39;-二氯乙烯）在这些过程中使用的是剧毒和致癌。查阅所有相关的材料安全数据表（MSDS）后才能使用。在进行实验时，请遵循适当的安全措施。 注:除非另有说明，开展在环境温度下的所有程序在100级层流罩。 1.偶极辅助Microchip的SPE的制作<ol><li> 聚甲基丙烯酸甲酯Microchip的制备 注意:在芯片的制造协议是相似的别处描述的8。 <ol><li>绘制芯片的网络图形（ 图1（a））使用根据制造商的协议计算机辅助设计（CAD）软件。 </li><li>装入的PMMA片材上的工作表（350毫米（长）×20毫米（宽）×2毫米（高））激光微加工系统，然后集中在PMMA片材的表面上的激光源。 </li><li>通过微加工系统的控制面板中选择在CAD软件打印 ，然后将力量 ， 速度和笔模式为45％（4.5 W），13％（99.06毫米秒-1），VECT。 注:参数，如力量，速度和笔模式影响了渠道的功能提前进行了调查。的评价方法相似，由元和达沙17在本研究中所选择的参数提议被用来机适应适当的信道，以导管无需复杂的学术目的。人们可以根据自己的需要选择激光加工的另一个条件。 </li><li>根据制造商的协议打印由该激光微加工系统的绘制图案，然后机聚甲基丙烯酸甲酯片。 图1（b） </强&gt;显示制造微芯片的布局图图1（c）显示该加工板的横截面的照片。 使用激光系统时避免因暴露于激光辐射严重的眼损伤注意！佩戴护目镜。适当的排气系统，是因为激光加工在生产烟雾/烟雾的建议。 </li><li>钻头三个1/16英寸的样品入口，缓冲器入口和在底板洗脱液入口和一个用于在盖板图1汇合出口直径接入孔（b）中 。 注意！避免在加工过程与钻头体接触，以防止人身伤害。戴手套钻孔时被禁止。 </li><li>通过10分钟的超声振荡器浸入加工板成1升的0.1％在1升烧杯（重量/体积）十二烷基硫酸钠（SDS）的溶液在搅拌。 </li><li>更换SDS水溶液W第i个去离子水，并通过10分钟的超声振荡器搅拌。 </li><li>用新的替换剩余的DI H 2 O和然后通过超声波振荡器浸入加工板在1L的去离子水中2 O搅拌10分钟。之后，干燥氮气的轻柔气流2分钟各清洗板。 </li><li>对齐用肉眼在两个加工板，然后通过使用粘合剂夹夹着两个玻璃板之间的两个板。 </li><li>接合在压缩下两个板在105℃下30分钟。 </li><li>酷三明治到环境温度，然后取出长尾夹和玻璃板。 </li><li>刀片1 / 16-英寸外径的聚（醚醚酮）（PEEK）管子插入通道孔，然后用双组分环氧系粘合剂固定导管。 </li><li>干燥在常温下的粘接剂12小时。 </li></ol></li><li> 聚甲基丙烯酸甲酯微芯片的通道内饰的修改 <br/&gt; 注:以下部分是指用细微的修改发表程序8,18,19 <ol><li>递送在100微升min的流速的饱和氢氧化钠（NaOH）溶液-通过蠕动泵1到微芯片12小时（72毫升总输送量）。 </li><li>除去残留的溶液中，然后冲洗用DI H 2澳通道内以100微升min的流速-经30分钟（3毫升总递送体积）蠕动泵1。 </li><li>除去残留的去离子H 2 O和然后在100微升min的流速提供0.5％（体积/体积）硝酸（HNO 3）溶液倒入微芯片-经由蠕动泵1 30分钟（3毫升总输送量）。 </li><li>除去残留的溶液中，然后传送一个50％（重量/体积）丙烯酰胺溶液到在100微升min的流速在黑暗中的微芯片- 1 VIA蠕动泵8小时（48毫升总交付量）。 </li><li>除去残留的溶液中，然后冲洗用DI H 2澳通道内以100微升min的流速-经30分钟（3毫升总递送体积）蠕动泵1。 </li><li>空气泵除去残留去离子水中2 O用蠕动泵再涂微芯片具有一个内部的内置光掩模允许提取通道的所需的区域进行曝光。 注:在内部建光掩模由黑色纸（114毫米（长）×22毫米（宽））包含一个开放的窗口（94毫米（长）×2 mm（宽））允许所需的区域提取通道的要暴露于光。 </li><li>在CL-SPE含形成溶液的制备<ol><li>冲洗用乙醇金额至少有三个墨盒的卷抑制剂去除固相萃取柱。 </li><li>冲洗墨盒1,1&#39;-二氯乙烯达在使用前至少有三个墨盒的卷。 </li><li>通过1毫升1,1&#39;-二氯乙烯的经处理过的暗盒，然后收集在一个样品瓶包裹在铝箔的级分（20ml）中。 </li><li>添加491微升1,1&#39;-二氯乙烯到含有12毫克的2,2&#39;-偶氮二异丁腈（AIBN），3.18毫升乙醇溶液中，并在100毫升的玻璃瓶1.65毫升己烷。 </li></ol></li><li>通过注射器注射填充与含氯-SPE形成溶液（约200微升）的芯片信道，然后暴露的微芯片，以紫外（UV 365）与365nm的最大发射波长照射10分钟（光强度〜2.65毫瓦厘米-2）。 注意！建议，因为紫外线照射在生产臭氧的适当的排气系统。 </li><li>更换新鲜含CL-SPE形成溶液（约200微升）用注射器仁济残留溶液ction并且把微芯片再次紫外线照射365 10分钟（光照强度〜2.65毫瓦厘米-2）。 </li><li>重复步骤1.2.9 18倍。 </li><li>通过30分钟（3毫升总投放量），蠕动泵1 -冲洗以100微升min的流速用乙醇通道内部。删除与蠕动泵的残留溶液后，存储在一个拉链袋为后续使用制造微芯片。 </li></ol></li></ol> 2.修改PMMA表面验证<ol><li> 接触角分析 <ol><li>切有机玻璃片（350毫米（长）×20毫米（宽）×2毫米（高））转换成聚甲基丙烯酸甲酯基底（50毫米（长）×20毫米（宽）×2毫米（高））的激光微加工系统。 </li><li>浸泡在饱和NaOH溶液40毫升聚甲基丙烯酸甲酯基底在50毫升锥形管，然后通过一个摇床上搅拌所得的混合物12小时。 </li><li>去除残留的溶液，然后用清水冲洗在P用40ml去离子H 2 O的MMA基板</li><li>沉浸在40毫升去离子水中2 O的PMMA基片，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的去离子H 2 O浸入的PMMA基板在40毫升0.5％（体积/体积）HNO 3溶液中，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的溶液。浸入的PMMA基板在40毫升的50％（重量/体积）丙烯酰胺溶液中，然后通过一个摇床上搅动所得的混合物在黑暗中8小时。 </li><li>去除残留的溶液中，然后冲洗用40毫升DI H 2 O的PMMA基板</li><li>浸入的PMMA基板在40毫升去离子水中2 O中，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的去离子H 2 O和然后干燥氮气的轻柔气流2分钟每PMMA衬底。 </li><li>编制CL-SPE含形成液<ol><li>冲洗用乙醇金额至少有三个墨盒的卷抑制剂去除固相萃取柱。 </li><li>同花顺1,1&#39;-二氯乙烯达在使用前至少三盒，卷盒。 </li><li>通过6毫升1,1&#39;-二氯乙烯的经处理过的暗盒，然后收集在一个样品瓶包裹在铝箔的级分（20ml）中。 </li><li>添加4.91毫升1,1&#39;-二氯乙烯到含有120毫克AIBN，31.8mmol毫升乙醇，并且在100-ml玻璃瓶16.5毫升己烷的溶液中。 </li></ol></li><li>敷2毫升含Cl--SPE形成溶液到PMMA基板的表面，然后暴露衬底于UV 365照射10分钟（光强度〜2.65毫瓦厘米-2）。 注意！建议，因为紫外线照射在生产臭氧的适当的排气系统。 </li><li>更换残留SOLU化用2ml新鲜的含氯-SPE形成溶液，然后暴露在基片再次UV 365照射10分钟（光强度〜2.65毫瓦厘米-2）。 </li><li>重复步骤2.1.12 18倍。 </li><li>除去残留的溶液中，然后在50毫升锥形管用40ml乙醇冲洗的PMMA基板。 </li><li>去除残留的溶液中，然后冲洗用40毫升DI H 2 O的PMMA基板</li><li>除去残留的去离子H 2 O和然后干燥氮气的轻柔气流2分钟每PMMA衬底。 </li><li>下降5微升去离子水中2 O到PMMA基材和通过根据制造商的协议的接触角计测定的接触角。 注意:使用三个重复测量值的平均值，以确定在每一种情况下报道的接触角。 </li></ol></li><li> 激光烧蚀（LA）-Inductively耦合等离子体质谱仪（ICP-MS）分析&lt;/ STRONG&gt; <ol><li>通过研钵和研杵研磨将8g PMMA珠成聚甲基丙烯酸甲酯粉末。 </li><li>浸泡在饱和NaOH溶液40毫升聚甲基丙烯酸甲酯粉末，在50-ml锥形管中，然后通过一个摇床上搅拌所得的混合物12小时。 </li><li>用5毫升的提示删除由数字吸管残留溶液，再用清水冲洗用40毫升DI H 2 O的PMMA粉末</li><li>沉浸在40毫升去离子水中2 O的聚甲基丙烯酸甲酯粉末，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的去离子H 2 O浸入聚甲基丙烯酸甲酯粉末在40毫升的0.5％（体积/体积）HNO 3溶液中，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的溶液。浸入聚甲基丙烯酸甲酯粉末在40毫升的50％（重量/体积）丙烯酰胺溶液中，然后通过一个摇床上搅动所得的混合物在黑暗中8小时。 </li><li>去除残留的溶液中，然后冲洗PMMA粉末机智ħ将40ml二H 2 O的</li><li>浸入聚甲基丙烯酸甲酯粉末在40毫升去离子水中2 O中，然后通过一个摇床上搅拌所得的混合物30分钟。 </li><li>除去残留的去离子H 2 O和然后在60℃下烘烤的PMMA粉末8小时。 </li><li>在CL-SPE含形成溶液的制备<ol><li>冲洗用乙醇金额至少有三个墨盒的卷抑制剂去除固相萃取柱。 </li><li>同花顺1,1&#39;-二氯乙烯达在使用前至少三盒，卷盒。 </li><li>通过16毫升1,1&#39;-二氯乙烯通过处理卡盒，然后收集在用铝箔包裹的试样小瓶（20毫升）的分数。 </li><li>添加14.73毫升1,1&#39;-二氯乙烯到含有360毫克AIBN 95.4毫升乙醇，和49.5毫升己烷在250毫升的玻璃瓶的溶液。 </li></ol></li><li>在6毫升含CL-SPE形成液混合PMMA粉末50-ml锥形管中，并同样转将1ml混合物从锥形管在24孔组织培养板六个孔中。 </li><li>封面用PMMA板组织培养板，然后暴露组织培养板在紫外线照射365 10分钟（光照强度〜2.65毫瓦厘米-2）。 注意！建议，因为紫外线照射在生产臭氧的适当的排气系统。 </li><li>替换用1ml的每个的新鲜含Cl--SPE形成溶液的残留溶液以及然后暴露组织培养板再次UV 365照射10分钟（光强度〜2.65毫瓦厘米-2）。 </li><li>重复步骤2.2.13 18倍。 </li><li>除去残留的溶液中，然后在每一个用1ml乙醇冲洗的PMMA粉末很好。 </li><li>除去残留的溶液中，然后在每一个1毫升的DI H 2 O冲洗的PMMA粉末以及</li><li>除去残留的去离子水中&lt;子&gt; 2 O，然后在60℃下烘烤的PMMA粉末8小时。 </li><li>经由液压机压缩干燥粉末（1克）为粒料，然后通过LA-ICP-MS系统测量为氯的信号。 注意:在m为氯的信号/ Z 35被选定为植入的C-Cl残基的一个指标。 一个193-nm激光用作消融源。 能源 ， 通量 ， 光斑尺寸 ，和重复率分别设定为75％，8.85Ĵ厘米-2，100微米，5赫兹。被要求至少有7个为每个结果重复测量。 LA-ICP-MS分析程序是指在别处发表的程序。20 </li></ol></li><li> 拉曼光谱分析 <ol><li>从步骤2.2.1执行协议步骤2.2.17。 </li><li>经由液压机压缩干燥粉末（1克）为粒料，然后用拉曼光谱仪采取的光谱。 注:使用100毫瓦的最大激光功率作为激发源的780纳米激光线。使用拉曼光谱范围从550至900厘米的区域-1调查的C-Cl残基，以聚甲基丙烯酸甲酯的附件。 </li></ol></li></ol> 3.偶极辅助SPE反应的表征<ol><li>从步骤2.2.1执行协议步骤2.2.17。 </li><li>在5毫升的20％（重量/体积）硝酸锰四水合物（锰（NO 3）2 4H 2 O） 的溶液浸入0.5克PMMA粉末，然后同样将所得的混合物用5毫升40毫马来酸盐缓冲溶液混合。 </li><li>通过使用纯的HNO 3水溶液调节所得混合物的pH至8，然后再通过一个摇床上搅拌混合物1小时。 </li><li>除去残留的溶液中，然后在60℃下烘烤的PMMA粉末8小时。在15-ml锥形管包裹在铝箔的X射线absor存储粉末ption近边结构（XANES）分析。 注:锰K边XANES谱用07A和国家同步辐射研究中心（NSRRC，台湾新竹）的光束线17C1收集。电子储存环用的1.5电子伏特的能量和100-200毫安的电流操作。的Si（​​111）双晶单色用于提供与第1能量高度单色光子束15千电子伏特和解析力（E /ΔE）高达5,000.The光子能量是通过使用公知的Mn的Mn标准物校准的K在6539.0电子伏特边吸收拐点。 6530和6,570伏特之间被用来研究对所提出的固相萃取反应表征偶极离子相互作用在该区域的Mn K边缘XANES光谱。 </li></ol>

<ol>
	<li>De Mello, A. J., Beard, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dealing+with+'real'+samples:+sample+pre-treatment+in+microfluidic+systems.">Dealing with 'real' samples: sample pre-treatment in microfluidic systems.</a> Lab Chip. 3 (1), 11-19 (2003).</li><li>Lichtenberg, J., de Rooij, N. F., Verpoorte, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sample+pretreatment+on+microfabricated+devices.">Sample pretreatment on microfabricated devices.</a> Talanta. 56 (2), 233-266 (2002).</li><li>Song, S., Singh, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On-chip+sample+preconcentration+for+integrated+microfluidic+analysis.">On-chip sample preconcentration for integrated microfluidic analysis.</a> Anal. Bioanal. Chem. 384 (1), 41-43 (2006).</li><li>Lafleur, J. P., Salin, E. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pre-concentration+of+trace+metals+on+centrifugal+microfluidic+discs+with+direct+determination+by+laser+ablation+inductively+coupled+plasma+mass+spectrometry.">Pre-concentration of trace metals on centrifugal microfluidic discs with direct determination by laser ablation inductively coupled plasma mass spectrometry.</a> J. Anal. At. Spectrom. 24 (11), 1511-1516 (2009).</li><li>Xue, S., Liu, Y., Li, H. -. F., Uchiyama, K., Lin, J. -. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+microscale+solid-phase+extraction+poly(dimethylsiloxane)+chip+for+enrichment+and+fluorescent+detection+of+metal+ions.">A microscale solid-phase extraction poly(dimethylsiloxane) chip for enrichment and fluorescent detection of metal ions.</a> Talanta. 116, 1005-1009 (2013).</li><li>Khoai, D. V., Kitano, A., Yamamoto, T., Ukita, Y., Takamura, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+high+sensitive+liquid+electrode+plasma-Atomic+emission+spectrometry+(LEP-AES)+integrated+with+solid+phase+pre-concentration.">Development of high sensitive liquid electrode plasma-Atomic emission spectrometry (LEP-AES) integrated with solid phase pre-concentration.</a> Microelectron. Eng. 111, 343-347 (2013).</li><li>Khoai, D. V., Yamamoto, T., Ukita, Y., Takamura, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=On-chip+solid+phase+extraction-liquid+electrode+plasma+atomic+emission+spectrometry+for+detection+of+trace+lead.">On-chip solid phase extraction-liquid electrode plasma atomic emission spectrometry for detection of trace lead.</a> Jpn. J. Appl. Phys. 53 (5S1), 1-5 (2014).</li><li>Shih, T. -. T., Chen, W. -. Y., Sun, Y. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Open-channel+chip-based+solid-phase+extraction+combined+with+inductively+coupled+plasma-mass+spectrometry+for+online+determination+of+trace+elements+in+volume-limited+saline+samples.">Open-channel chip-based solid-phase extraction combined with inductively coupled plasma-mass spectrometry for online determination of trace elements in volume-limited saline samples.</a> J. Chromatogr. A. 1218 (16), 2342-2348 (2011).</li><li>Chen, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+solid+phase+microextraction+on+a+microchip+combined+with+electrothermal+vaporization-inductively+coupled+plasma+mass+spectrometry+for+determination+of+Cd,+Hg+and+Pb+in+cells.">Magnetic solid phase microextraction on a microchip combined with electrothermal vaporization-inductively coupled plasma mass spectrometry for determination of Cd, Hg and Pb in cells.</a> J. Anal. At. Spectrom. 25 (12), 1931-1938 (2010).</li><li>Kim, Y. H., Kim, G. Y., Lim, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Micro+Pre-concentration+and+Separation+of+Metal+Ions+Using+Microchip+Column+Packed+with+Magnetic+Particles+Immobilized+by+Aminobenzyl+Ethylenediaminetetraacetic+Acid.">Micro Pre-concentration and Separation of Metal Ions Using Microchip Column Packed with Magnetic Particles Immobilized by Aminobenzyl Ethylenediaminetetraacetic Acid.</a> Bull. Korean Chem. Soc. 31 (4), 905-909 (2010).</li><li>Strabburg, S., Wollenweber, D., Wunsch, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contamination+caused+by+ion-exchange+resin!?+Consequences+for+ultra-trace+analysis.">Contamination caused by ion-exchange resin!? Consequences for ultra-trace analysis.</a> Fresenius J. Anal. Chem. 360 (7-8), 792-794 (1998).</li><li>Watts, J. F., Chehimi, M. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X-ray+Photoelectron+Spectroscopy+Investigations+of+Acid-Base+Interactions+in+Adhesion.">X-ray Photoelectron Spectroscopy Investigations of Acid-Base Interactions in Adhesion.</a> J. Adhes. 41 (1-4), 81-91 (1993).</li><li>Eboatu, A. N., Diete-Spiff, S. T., Ezenweke, L. O., Omalu, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Use+of+Polymers+as+Sequestering+Agents+for+Toxic+Metal+Ions.">The Use of Polymers as Sequestering Agents for Toxic Metal Ions.</a> J. Appl. Polym. Sci. 85 (13), 2781-2786 (2002).</li><li>Fiorini, G. S., Chiu, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Disposable+microfluidic+devices:+fabrication,+function,+and+application.">Disposable microfluidic devices: fabrication, function, and application.</a> Biotechniques. 38 (3), 429-446 (2005).</li><li>Shadpour, H., Musyimi, H., Chen, J., Soper, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiochemical+properties+of+various+polymer+substrates+and+their+effects+on+microchip+electrophoresis+performance.">Physiochemical properties of various polymer substrates and their effects on microchip electrophoresis performance.</a> J. Chromatogr. A. 1111 (2), 238-251 (2006).</li><li>Shih, T. -. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+dipole-assisted+solid-phase+extraction+microchip+combined+with+inductively+coupled+plasma-mass+spectrometry+for+online+determination+of+trace+heavy+metals+in+natural+water.">A dipole-assisted solid-phase extraction microchip combined with inductively coupled plasma-mass spectrometry for online determination of trace heavy metals in natural water.</a> Analyst. 140 (2), 600-608 (2015).</li><li>Yuan, D., Das, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+and+theoretical+analysis+of+direct-write+laser+micromachining+of+polymethyl+methacrylate+by+CO2+laser+ablation.">Experimental and theoretical analysis of direct-write laser micromachining of polymethyl methacrylate by CO2 laser ablation.</a> J. Appl. Phys. 101 (2), 1-6 (2007).</li><li>Xie, S., Svec, F., Frechet, J. M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+Porous+Hydrophilic+Monoliths:+Effect+of+the+Polymerization+Conditions+on+the+Porous+Properties+of+Poly(acrylamide-co-N,N'-methylenebisacrylamide)+Monolithic+Rods.">Preparation of Porous Hydrophilic Monoliths: Effect of the Polymerization Conditions on the Porous Properties of Poly(acrylamide-co-N,N'-methylenebisacrylamide) Monolithic Rods.</a> J. Polym. Sci. Pol. Chem. 35 (6), 1013-1021 (1997).</li><li>Ericson, C., Liao, J. -. L., Nakazato, K., Hjerten, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+of+continuous+beds+for+electrochromatography+and+reversed-phase+liquid+chromatography+of+low-molecular-mass+compounds.">Preparation of continuous beds for electrochromatography and reversed-phase liquid chromatography of low-molecular-mass compounds.</a> J. Chromatogr. A. 767 (1-2), 33-41 (1997).</li><li>Shih, T. -. T., Lin, C. -. H., Hsu, I. -. H., Chen, J. -. Y., Sun, Y. -. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+Titanium+Dioxide-Coated+Microfluidic-Based+Photocatalyst-Assisted+Reduction+Device+to+Couple+High-Performance+Liquid+Chromatography+with+Inductively+Coupled+Plasma-Mass+Spectrometry+for+Determination+of+Inorganic+Selenium+Species.">Development of a Titanium Dioxide-Coated Microfluidic-Based Photocatalyst-Assisted Reduction Device to Couple High-Performance Liquid Chromatography with Inductively Coupled Plasma-Mass Spectrometry for Determination of Inorganic Selenium Species.</a> Anal. Chem. 85 (21), 10091-10098 (2013).</li><li>Carre, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Polar+interactions+at+liquid/polymer+interfaces.">Polar interactions at liquid/polymer interfaces.</a> J. Adhes. Sci. Technol. 21 (10), 961-981 (2007).</li><li>Willis, H. A., Zichy, V. J. I., Hendra, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Laser-Raman+and+Infra-red+Spectra+of+Poly(Methyl+Methacrylate).">The Laser-Raman and Infra-red Spectra of Poly(Methyl Methacrylate).</a> Polymer. 10, 737-746 (1969).</li><li>Hendra, P. J., Mwkenzie, J. R., Holliday, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+laser-Raman+spectrum+of+polyvinylidene+chloride.">The laser-Raman spectrum of polyvinylidene chloride.</a> Spectroc. Acta Pt. A-Molec. Biomolec. Spectr. 25 (8), 1349-1354 (1969).</li></ol>

The authors have nothing to disclose.

为偶极辅助SPE微芯片的制备的详细程序上面作了介绍。在本节中，修改协议的关于在PMMA的C-Cl残基和含氯-PMMA，将其用作用于痕量金属离子的测定提取介质的可行性的植入的效用，是评估一步一步的。对于表面验证目的，样品类型是其与分析仪器的兼容性的基础上选择的。换句话说，该类型通过类似的方法制备的测试样品在根据分析仪器的要求进行了测定。例如，使用一个衬底型样品的接触角的测定，而被用于对LA-ICP-MS，拉曼分光的粉末包装型样品，和XANES分析。 最初，以监测由所述化学官能武官发生变化所提出的程序期间d，来聚甲基丙烯酸甲酯的表面上，对应于每个步骤进行所得产物的接触角分析（ 图2）。如显示在图2中，接触角的变化清楚地表明，在修改的程序发生表面的变化，而 ​​测定该最终产物是在协议与先前报告的结果为80.3°±0.43°的接触角。21 此外，关于改性的PMMA的C-Cl残基的存在通过LA-ICP-MS分析也证实。与由烧蚀天然PMMA获得的结果相比，被烧蚀具有C-Cl残基改性聚甲基丙烯酸甲酯expectably观察氯不同信号（ 图3的（a））。 拉曼光谱收集用于进一步验证的C-Cl残基，以聚甲基丙烯酸甲酯的附件。如图Figu再如图3（b），682 -1和718 -1中的改性的PMMA的光谱中观察到与四氯化碳2不对称伸缩振动有关的两个特征峰，并与由Willis 等人报道的结果，在合理的良好的一致性22和亨德拉等人 23换言之，该C-Cl残基，以聚甲基丙烯酸甲酯的附件可以成功地修改后实现的。 此外，为了澄清这项研究提出了萃取机理，被采用的XANES分析。如在图4所示，高度电负性的C-Cl残基和带正电的金属离子之间的相互作用可以由主导吸收边缘在对应于与锰2+离子处理的改性的PMMA的XANES光谱的存在来确认。因此，偶极 - 静电相互作用将确实应用到TR片上提取王牌金属分析。在台湾从两江收集的水样品的详细分析结果已在其他地方描述。16 据我们所知，这是利用一种创新的工作战略的痕量金属离子的测定芯片上SPE反应的第一次尝试，并与其他芯片SPE技术相比，开发的设备是显著耐用（ 即 160多个作品的分析可以不显著恶化的提取效率方面实现）。然而，因为这样的取出机构主要依赖于高度电负性的C-Cl残基和带正电的金属离子之间的相互作用，所提出的技术预计是不适于带负电荷的物质的提取为止。

从环境管理和污染防治的观点，微量金属污染严重或毒性问题是一个世界性的问题。芯片样品前处理技术已被广泛接受为重点，以处理和分析通过基于芯片的平台实际样品的成功，因为原料样品中意外并存的化学物质往往妨碍痕迹量存在分析物的准确测定一个适当的1在现有的技术中，芯片上的固相萃取（SPE）是特别受欢迎的痕量金属分析，因为该技术允许样品净化，也可以同时进行分析物富集是用于从复杂的盐基质金属离子的隔离非常有用的。 2,3 片上SPE技术用于痕量元素测定的进步一直在稳步发展。在创业初期，T他SPE通过加载市售的树脂进入微通道构造树脂填充的SPE单元制备芯片。4-7这偶尔需要的分析物进行衍生，以使金属离子转变成树脂保持性形式4的另一种方法用于基于芯片的SPE装置的制备是利用芯片信道作为SPE吸附剂微量金属的简单表面改性后的收集。8最近几年，涉及磁性纳米颗粒（的MNP）和特定化学物质的掺入一种新兴趋势包含能够金属离子的有效保留的官能团。在对比的商业树脂，所述的MNP被修改以如后它们被包装到微通道与外部磁场的t借助于γ巯基（γ-MPTS）9和氨基苄乙二胺四乙酸（ABEDTA）10化合物Ø实现金属离子的选择性提取。 虽然在芯片上SPE技术的发展显著的进步已经见证，报道的技术通常功能的基础上无论是离子交换或螯合作用。使用的技术，如这些具有需要不可避免操作程序，包括那些有空调，洗衣机，或再生相关联，以保持所述分析性能的缺点。不幸的是，需要额外的操作程序，不仅扩展了对每个分析所需要的时间，而且造成的风险高空白值和复制的结果。11因此，对于芯片SPE技术合作战略的选择是当务之急痕量金属分析。 1993年，瓦特和Chehimi 12发现，金属离子具有朝向聚合材料滞留倾向，多数分析物有效地保留在氯（Cl）的-containi纳克聚合物材料，聚（氯乙烯）（PVC），除了钠离子。因此，在2002年，Eboatu 等 13还对一些有毒金属从PVC的解决方案封存的报道。因为这表明含CL-的表现分析物富集和盐基体消除优异性能的聚合物材料，用CL-含SPE功能的基于芯片的装置被认为是一种新型的芯片固相萃取技术的发展，为确定一个有吸引力的战略的微量金属离子。考虑材料的特性，如易于制造的，希望的化学/机械特性，并且光学透明度，14,15这项研究了聚甲基丙烯酸甲酯（PMMA）的优点来制造微型装置。然后，将含有氯-SPE的功能植入用于痕量金属离子的测定的新颖的片SPE技术的发展所制造的装置。16 - [Remarkably，关于在信道内的高度电负性的C-Cl残基和带正电的金属离子之间的偶极子 - 离子相互作用的创新提取机构的依赖，使得有可能避免在一般芯片上的SPE程序所采取的措施，从而导致任污染显着减少因使用过量的试剂或归因于额外的步骤的劳力。在这方面的贡献提供的协议将使来自不同背景的研究人员制造了他们的工作偶极辅助SPE微芯片。对于所制造的微芯片详细的表征程序被描述为良好。

<table border="0" cellpadding="0" cellspacing="0" width="1311">
	<tbody>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">AutoCAD</td>
			<td style="width:231px;">Autodesk</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">http://www.autodesk.com/education/free-software/autocad</td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;">Poly(methyl methacrylate) (PMMA) sheet</td>
			<td style="width:231px;">Kun Quan Engineering Plastics</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">350 mm (L) x 20 mm (W) x 2 mm (H). The glass transition temperature (Tg) of PMMA sheets is ranged from 102&#8211;110 &#176;C. The UV transmittance of the PMMA at 365 nm is 91.2%.</td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;width:383px;">Micromachining system</td>
			<td style="width:231px;">Laser Life</td>
			<td>LES-10</td>
			<td style="width:423px;">Maximum laser power: 10 W. Maximium engraving speed: 762 mm s&#8722;1.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">High-resolution optical microscope</td>
			<td style="width:231px;">Ching Hsing Computer-Tech</td>
			<td style="width:276px;">FS-230</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Power Image Analysis system (PIA)</td>
			<td style="width:231px;">Ching Hsing Computer-Tech</td>
			<td style="width:276px;">PIA V16.1</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Multi drilling machines</td>
			<td style="width:231px;">N/A</td>
			<td>LT-848</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Deionized water (D. I. H2O)</td>
			<td style="width:231px;">Millipore</td>
			<td>Milli-Q Integral 5 System</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Sodium dodecyl sulfate (SDS)</td>
			<td style="width:231px;">J. T. Baker</td>
			<td>4095-04</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Ultrasound oscillator</td>
			<td style="width:231px;">Elma</td>
			<td>Transsonic Digital</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Glass board</td>
			<td style="width:231px;">N/A</td>
			<td style="width:276px;">N/A</td>
			<td>160 mm (L) x 35 mm (W) x 2 mm (H); fragile</td>
		</tr>
		<tr height="70">
			<td height="70" style="height:70px;">Binder clip</td>
			<td style="width:231px;">SDI</td>
			<td style="width:276px;">0234T-1</td>
			<td style="width:423px;">http://stationery.sdi.com.tw/product_detail.php?Key=322&#38;cID=55&#38;uID=6</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Precision oven</td>
			<td style="width:231px;">Yeong Shin</td>
			<td style="width:276px;">DK-45</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;">Poly(etheretherketone) (PEEK) tube</td>
			<td style="width:231px;">VICI</td>
			<td style="width:276px;">JR-T-6002 (0.5 mm i.d.); JR-T-6001 (0.25 mm i.d.)</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Polymer tubing&#160; cutter</td>
			<td style="width:231px;">Upchurch Scientific</td>
			<td style="width:276px;">A-327</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:53px;">Two-component epoxy-based adhesive</td>
			<td style="width:231px;">Richwang</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">Skin irritative. The major components are an epoxy resin and a hardener.</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Peristaltic pump</td>
			<td style="width:231px;">Gilson</td>
			<td>Minipuls 3</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Peristaltic tube</td>
			<td style="width:231px;">Gilson</td>
			<td>F117934</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Sodium hydroxide (NaOH)</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td style="width:276px;">30620</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Nitric acid (HNO3)</td>
			<td style="width:231px;">J. T. Baker</td>
			<td style="width:276px;">959834</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Acrylamide (prop-2-enamide, C3H5NO)</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td style="width:276px;">A8887</td>
			<td>Acutely toxic and carcinogenic</td>
		</tr>
		<tr height="78">
			<td height="78" style="height:78px;width:383px;">In-house-built photomask</td>
			<td style="width:231px;">N/A</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">The in-house-built photomask was made of a black paper (114 mm (L) &#215; 22 mm (W)) that contained an open window (94 mm (L) &#215; 2 mm (W)) allowing the desired region</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">1,1-Dichloroethylene</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td style="width:276px;">163032</td>
			<td>Acutely toxic and carcinogenic</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Cartridge</td>
			<td style="width:231px;">Dikma</td>
			<td style="width:276px;">ProElut AL-B&#160;</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">2,2-Azobisisobutyronitrile (AIBN, C8H12N4)</td>
			<td style="width:231px;">Showa Chemical&#160;</td>
			<td style="width:276px;">0159-2130</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Ethanol</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td>32221</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Hexanes (C6H14)</td>
			<td style="width:231px;">Millinckrodt Chemical</td>
			<td style="width:276px;">5189-08</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;width:383px;">&#160;In-house-built irradiation system</td>
			<td style="width:231px;">Great Lighting (UV-A lamp)</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">An opaque box with an UV-A lamp (40 W, maximum emission at 365 nm)</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Glass vial</td>
			<td style="width:231px;">Yeong Shin</td>
			<td style="width:276px;">132300019</td>
			<td>Fragile</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Aluminum foil</td>
			<td style="width:231px;">Diamond</td>
			<td style="width:276px;">N/A</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;">Conical tubes with screw caps</td>
			<td style="width:231px;">labcon</td>
			<td style="width:276px;">3181-345-008 (50 mL); 3131-345-008 (15 mL)</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Rocking shaker</td>
			<td style="width:231px;">TKS</td>
			<td style="width:276px;">RS-01</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Contact angle meter</td>
			<td style="width:231px;">First Ten Angstroms</td>
			<td>FTA 125</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">PMMA bead</td>
			<td style="width:231px;">Scientific Polymer Products</td>
			<td style="width:276px;">037A</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Mortar and pestle, agate&#160;</td>
			<td style="width:231px;">Yeong Shin</td>
			<td style="width:276px;">139000004</td>
			<td>Fragile</td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;">Tissue culture plate</td>
			<td style="width:231px;">AdvanGene Life Science Plasticware</td>
			<td style="width:276px;">AGC-CP-24S-50EA</td>
			<td>24-Well, non-treated, sterilized</td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Hydraulic press</td>
			<td style="width:231px;">Panchum</td>
			<td style="width:276px;">Press-200</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Laser ablation</td>
			<td style="width:231px;">New Wave Research</td>
			<td>NWR193</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:26px;width:383px;">Inductively coupled plasma-mass spectrometer</td>
			<td style="width:231px;">Agilent Technologies</td>
			<td>Agilent 7500a</td>
			<td></td>
		</tr>
		<tr height="52">
			<td height="52" style="height:52px;width:383px;">Glass bottle</td>
			<td style="width:231px;">DURAN</td>
			<td style="width:276px;">21801245 (100 mL); 21801365 (250 mL)</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Dispersive Raman spectrometer</td>
			<td style="width:231px;">Thermo Fisher Scientific</td>
			<td>Nicolet Almega XR</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Manganese nitrate tetrahydrate (Mn(NO3)2&#215;4H2O)</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td style="width:276px;">63547</td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;width:383px;">Maleic acid disodium salt hydrate (C4H4Na2O5)</td>
			<td style="width:231px;">Sigma&#8211;Aldrich</td>
			<td style="width:276px;">M9009</td>
			<td></td>
		</tr>
		<tr height="78">
			<td height="78" style="height:78px;width:383px;">X-ray absorption near edge structure (XANES)&#160;</td>
			<td style="width:231px;">N/A</td>
			<td style="width:276px;">N/A</td>
			<td style="width:423px;">The Mn K-edge XANES analyses were conducted at 07A and 17C1 beamlines of the National Synchrotron Radiation Research Center (NSRRC) in Taiwan.</td>
		</tr>
	</tbody>
</table>

fabrication of a dipole-assisted solid phase extraction microchip for trace metal analysis in water samples

本文介绍了可用于水样中痕量金属分析偶极辅助固相萃取（SPE）微芯片的制造协议。提供的基于芯片的SPE技术演进的简要概述。这之后是介绍特定的聚合物材料以及它们在SPE作用。开发一个创新的偶极辅助SPE技术中，含SPE功能的氯（Cl）溶液注入到聚（甲基丙烯酸甲酯）（PMMA）的微芯片。这里，不同的分析技术，包括接触角分析，拉曼光谱分析中，以及激光烧蚀电感耦合等离子体 - 质谱（LA-ICP-MS）的分析被雇用来验证对的C- Cl残基的植入协议的效用PMMA。的X射线吸收近边结构的分析结果（XANES）分析还凭借偶极的证实用作萃取介质中的含氯-PMMA的可行性高电负性的C-Cl残基和带正电的金属离子之间的离子相互作用。

图2描述了在聚甲基丙烯酸甲酯微芯片的流路变更手续发生反应。采用接触角分析，监测建议的程序在表面的变化。一个LA-ICP-MS系统和色散型拉曼光谱仪被雇用来验证C-C1的成功修饰部分的PMMA基板上形成（ 如图3（a），（b））的 。所提出的偶极辅助SPE反应的特点是XANES分析（ 图4）。 <img alt="图1" src="/files/ftp_upload/53500/53500fig1.jpg" /> 图1:PMMA微芯片。（ 一 ）为了制造微芯片的模式文件的快照。 （ 二 ）所制造的微芯片的布局:S，E和B代表的引入端口，用于将样品，洗脱剂，和BUF外汇储备的解决方案，分别为; O代表出口。黑色圆圈表示每个所钻的进入孔。用于引入样品和缓冲溶液的通道都形成与提取通道30°的角度。有效提取通道，其被定义为从样品和缓冲液至汇合出口的流动的会聚点的距离的长度，是94毫米。 （c）该加工板的横截面的照片。从参考转载。 16通过化学的英国皇家学会的许可。 <a href="https://www.jove.com/files/ftp_upload/53500/53500fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/53500/53500fig2.jpg" /> 图2. 方案的PMMA微通道修改。插图photogrAPHS显示对应于序列中的所得产物的接触角。接触角是通过使用一水滴的图像来确定。被用于确定在每一种情况下报道的接触角三个重复测量的平均值。从参考转载。 16通过化学的英国皇家学会的许可。 <a href="https://www.jove.com/files/ftp_upload/53500/53500fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/53500/53500fig3.jpg" /> 图3. 表面PMMA修饰的验证。（a）由消融二者的PMMA和聚甲基丙烯酸甲酯与C-Cl残基改性得到的信号为氯。插图示出对应于每个获得的信号的消融位置。 （ 二 ）原生拉曼光谱和修改PMMA。从参考转载。 16由许可皇家化学学会。 <a href="https://www.jove.com/files/ftp_upload/53500/53500fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/53500/53500fig4.jpg" /> 图4. 锰K边XANES与锰 离子 离子 处理改性PMMA和PMMA修饰谱 ，修饰的PMMA的光谱是作为红线。显示的吸收光谱修饰PMMA的高度负电的C-Cl残基和锰离子的离子之间的相互作用是作为蓝线。从参考转载。 16通过化学的英国皇家学会的许可。 <a href="https://www.jove.com/files/ftp_upload/53500/53500fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a>

Watch this Scientific Journal Video about Fabrication of a Dipole-assisted Solid Phase Extraction Microchip for Trace Metal Analysis in Water Samples at JoVE.com

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

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 ...

fabrication-dipole-assisted-solid-phase-extraction-microchip-for

Research

Education

JoVE Core

JoVE Journal

Physics

Bioengineering

Unit 1

Electric Charges and Fields

SCIENTISTS IN ACTION

8.6K 次观看.  National Tsing Hua University. The fabrication protocol of a dipole-assisted solid phase extraction microchip for the trace metal analysis is presented.

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

Fabrication of Electrochemical-DNA Biosensors for the Reagentless Detection of Nucleic Acids, Proteins and Small Molecules

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. Many patients would be better served by rapid, bedside tests. To this end our laboratory and others have developed a versatile, reagentless 
biosensor platform that supports the quantitative, reagentless, electrochemical detection of nucleic acids (DNA, RNA), proteins (including antibodies) and small 
molecules analytes directly in unprocessed clinical and environmental samples. In this video, we demonstrate the preparation and use of several biosensors in this 
"E-DNA" class. In particular, we fabricate and demonstrate sensors for the detection of a target DNA sequence in a polymerase chain reaction mixture, an HIV-specific antibody and the drug cocaine. The preparation procedure requires only three hours of hands-on effort followed by an overnight incubation, and their use requires only minutes.

"E-DNA" sensors, reagentless, electrochemical biosensors that perform well even when challenged directly in blood and other complex matrices, have been adapted to the detection of a wide range of nucleic acid, protein and small molecule analytes. Here we present a general procedure for the fabrication and use of such sensors.

无试剂检测核酸，蛋白质和小分子的电化学DNA生物传感器的制备

As medicine is currently practiced, doctors send specimens to a central laboratory for testing and thus must wait hours or days to 
receive the results. ...

科研

生物工程

Creating Two-Dimensional Patterned Substrates for Protein and Cell Confinement

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1 While microcontact printing can be employed to directly print a large number of molecules, including proteins,2 DNA,3 and silanes,4 the formation of self-assembled monolayers (SAMs) from long chain alkane thiols on gold provides a simple way to confine proteins and cells to specific patterns containing adhesive and resistant regions. This confinement can be used to control cell morphology and is useful for examining a variety of questions in protein and cell biology. Here, we describe a general method for the creation of well-defined protein patterns for cellular studies.5 This process involves three steps: the production of a patterned master using photolithography, the creation of a PDMS stamp, and microcontact printing of a gold-coated substrate. Once patterned, these cell culture substrates are capable of confining proteins and/or cells (primary cells or cell lines) to the pattern.
The use of self-assembled monolayer chemistry allows for precise control over the patterned protein/cell adhesive regions and non-adhesive regions; this cannot be achieved using direct protein stamping. Hexadecanethiol, the long chain alkane thiol used in the microcontact printing step, produces a hydrophobic surface that readily adsorbs protein from solution. The glycol-terminated thiol, used for backfilling the non-printed regions of the substrate, creates a monolayer that is resistant to protein adsorption and therefore cell growth.6 These thiol monomers produce highly structured monolayers that precisely define regions of the substrate that can support protein adsorption and cell growth. As a result, these substrates are useful for a wide variety of applications from the study of intercellular behavior7 to the creation of microelectronics.8
While other types of monolayer chemistry have been used for cell culture studies, including work from our group using trichlorosilanes to create patterns directly on glass substrates,9 patterned monolayers formed from alkane thiols on gold are straight-forward to prepare. Moreover, the monomers used for monolayer preparation are commercially available, stable, and do not require storage or handling under inert atmosphere. Patterned substrates prepared from alkane thiols can also be recycled and reused several times, maintaining cell confinement.10

Self-assembled monolayers (SAMs) formed from long chain alkane thiols on gold provide well-defined substrates for the formation of protein patterns and cell confinement. Microcontact printing of hexadecanethiol using a polydimethylsiloxane (PDMS) stamp followed by backfilling with a glycol-terminated alkane thiol monomer produces a pattern where protein and cells adsorb only to the stamped hexadecanethiol region.

蛋白质和细胞坐月子创建二维图案衬底

Microcontact printing provides a rapid, highly reproducible method for the creation of well-defined patterned substrates.1  While microcontact printing ...

Solid-phase Submonomer Synthesis of Peptoid Polymers and their Self-Assembly into Highly-Ordered Nanosheets

Peptoids are a novel class of biomimetic, non-natural, sequence-specific heteropolymers that resist proteolysis, exhibit potent biological activity, and fold into higher order nanostructures. Structurally similar to peptides, peptoids are poly N-substituted glycines, where the side chains are attached to the nitrogen rather than the alpha-carbon. Their ease of synthesis and structural diversity allows testing of basic design principles to drive de novo design and engineering of new biologically-active and nanostructured materials.
Here, a simple manual peptoid synthesis protocol is presented that allows the synthesis of long chain polypeptoids ( up to 50mers) in excellent yields. Only basic equipment, simple techniques (e.g. liquid transfer, filtration), and commercially available reagents are required, making peptoids an accessible addition to many researchers' toolkits. The peptoid backbone is grown one monomer at a time via the submonomer method which consists of a two-step monomer addition cycle: acylation and displacement. First, bromoacetic acid activated in situ with N,N'-diisopropylcarbodiimide acylates a resin-bound secondary amine. Second, nucleophilic displacement of the bromide by a primary amine follows to introduce the side chain. The two-step cycle is iterated until the desired chain length is reached. The coupling efficiency of this two-step cycle routinely exceeds 98% and enables the synthesis of peptoids as long as 50 residues. Highly tunable, precise and chemically diverse sequences are achievable with the submonomer method as hundreds of readily available primary amines can be directly incorporated.
 
Peptoids are emerging as a versatile biomimetic material for nanobioscience research because of their synthetic flexibility, robustness, and ordering at the atomic level. The folding of a single-chain, amphiphilic, information-rich polypeptoid into a highly-ordered nanosheet was recently demonstrated. This peptoid is a 36-mer that consists of only three different commercially available monomers: hydrophobic, cationic and anionic. The hydrophobic phenylethyl side chains are buried in the nanosheet core whereas the ionic amine and carboxyl side chains align on the hydrophilic faces. The peptoid nanosheets serve as a potential platform for membrane mimetics, protein mimetics, device fabrication, and sensors. Methods for peptoid synthesis, sheet formation, and microscopy imaging are described and provide a simple method to enable future peptoid nanosheet designs.

A simple and general manual peptoid synthesis method involving basic equipment and commercially available reagents is outlined, enabling peptoids to be easily synthesized in most laboratories. The synthesis, purification and characterization of an amphiphilic peptoid 36mer is described, as well as its self-assembly into highly-ordered nanosheets.

固相Submonomer Peptoid聚合物及其自组装成高度有序的纳米片的合成

Peptoids are a novel class of biomimetic, non-natural, sequence-specific heteropolymers that resist proteolysis, exhibit potent biological activity, and ...

Cellular Lipid Extraction for Targeted Stable Isotope Dilution Liquid Chromatography-Mass Spectrometry Analysis

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including numerous stereoisomers1,2. These metabolites can be formed from free or esterified fatty acids. Many of these oxidized metabolites have biological activity and have been implicated in various diseases including cardiovascular and neurodegenerative diseases, asthma, and cancer3-7. Oxidized bioactive lipids can be formed enzymatically or by reactive oxygen species (ROS). Enzymes that metabolize fatty acids include cyclooxygenase (COX), lipoxygenase (LO), and cytochromes P450 (CYPs)1,8. Enzymatic metabolism results in enantioselective formation whereas ROS oxidation results in the racemic formation of products.
While this protocol focuses primarily on the analysis of AA- and some LA-derived bioactive metabolites; it could be easily applied to metabolites of other fatty acids. Bioactive lipids are extracted from cell lysate or media using liquid-liquid (l-l) extraction. At the beginning of the l-l extraction process, stable isotope internal standards are added to account for errors during sample preparation. Stable isotope dilution (SID) also accounts for any differences, such as ion suppression, that metabolites may experience during the mass spectrometry (MS) analysis9. After the extraction, derivatization with an electron capture (EC) reagent, pentafluorylbenzyl bromide (PFB) is employed to increase detection sensitivity10,11. Multiple reaction monitoring (MRM) is used to increase the selectivity of the MS analysis. Before MS analysis, lipids are separated using chiral normal phase high performance liquid chromatography (HPLC). The HPLC conditions are optimized to separate the enantiomers and various stereoisomers of the monitored lipids12. This specific LC-MS method monitors prostaglandins (PGs), isoprostanes (isoPs), hydroxyeicosatetraenoic acids (HETEs), hydroxyoctadecadienoic acids (HODEs), oxoeicosatetraenoic acids (oxoETEs) and oxooctadecadienoic acids (oxoODEs); however, the HPLC and MS parameters can be optimized to include any fatty acid metabolites13.
Most of the currently available bioanalytical methods do not take into account the separate quantification of enantiomers. This is extremely important when trying to deduce whether or not the metabolites were formed enzymatically or by ROS. Additionally, the ratios of the enantiomers may provide evidence for a specific enzymatic pathway of formation. The use of SID allows for accurate quantification of metabolites and accounts for any sample loss during preparation as well as the differences experienced during ionization. Using the PFB electron capture reagent increases the sensitivity of detection by two orders of magnitude over conventional APCI methods. Overall, this method, SID-LC-EC-atmospheric pressure chemical ionization APCI-MRM/MS, is one of the most sensitive, selective, and accurate methods of quantification for bioactive lipids.

This protocol will demonstrate the extraction and analysis of free and esterified bioactive fatty acids from cells. Fatty acids are accurately quantified using stable isotope dilution, chiral liquid chromatography, electron capture atmospheric chemical ionization multiple reaction monitoring mass spectrometry (SID-LC-ECAPCI-MRM/MS).

有针对性的稳定同位素稀释液相色谱 - 质谱法分析细胞脂质提取

The metabolism of fatty acids, such as arachidonic acid (AA) and linoleic acid (LA), results in the formation of oxidized bioactive lipids, including ...

Microfluidic Fabrication of Polymeric and Biohybrid Fibers with Predesigned Size and Shape

A &#8220;sheath&#8221; fluid passing through a microfluidic channel at low Reynolds number can be directed around another &#8220;core&#8221; stream and used to dictate the shape as well as the diameter of a core stream.&#160;Grooves in the top and bottom of a microfluidic channel were designed to direct the sheath fluid and shape the core fluid.&#160;By matching the viscosity and hydrophilicity of the sheath and core fluids, the interfacial effects are minimized and complex fluid shapes can be formed.&#160;Controlling the relative flow rates of the sheath and core fluids determines the cross-sectional area of the core fluid.&#160;Fibers have been produced with sizes ranging from 300 nm to ~1 mm, and fiber cross-sections can be round, flat, square, or complex as in the case with double anchor fibers. Polymerization of the core fluid downstream from the shaping region solidifies the fibers.&#160;Photoinitiated click chemistries are well suited for rapid polymerization of the core fluid by irradiation with ultraviolet light.&#160;Fibers with a wide variety of shapes have been produced from a list of polymers including liquid crystals, poly(methylmethacrylate), thiol-ene and thiol-yne resins, polyethylene glycol, and hydrogel derivatives. Minimal shear during the shaping process and mild polymerization conditions also makes the fabrication process well suited for encapsulation of cells and other biological components.

Two adjacent fluids passing through a grooved microfluidic channel can be directed to form a sheath around a prepolymer core; thereby&#160;determining both shape and cross-section. Photoinitiated polymerization, such as thiol click chemistry, is well suited for rapidly solidifying the core fluid into a microfiber with predetermined size and shape.

预设计尺寸和形状的聚合物纤维和生物贿赂纤维的微流体制造

A &#8220;sheath&#8221; fluid passing through a microfluidic channel at low Reynolds number can be directed around another &#8220;core&#8221; stream and ...

Fabrication of Mechanically Tunable and Bioactive Metal Scaffolds for Biomedical Applications

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness and low bioactivity of metals. In this study, we have developed a new method towards fabricating a new type of bioactive and mechanically reliable porous metal scaffolds-densified porous Ti scaffolds. The method consists of two fabrication processes, 1) the fabrication of porous Ti scaffolds by dynamic freeze casting, and 2) coating and densification of the porous scaffolds. The dynamic freeze casting method to fabricate porous Ti scaffolds allowed the densification of porous scaffolds by minimizing the chemical contamination and structural defects. The densification process is distinctive for three reasons. First, the densification process is simple, because it requires a control of only one parameter (degree of densification). Second, it is effective, as it achieves mechanical enhancement and sustainable release of biomolecules from porous scaffolds. Third, it has broad applications, as it is also applicable to the fabrication of functionally graded porous scaffolds by spatially varied strain during densification.

Bioactive and mechanically reliable metal scaffolds have been fabricated through a method which consists of two processes, dynamic freeze casting for the fabrication of porous Ti, and coating and densification of the Ti scaffolds. The densification process is simple, effective and applicable to the fabrication of functionally graded scaffolds.

加工机械可调和生物活性金属支架的生物医学应用

Biometal systems have been widely used for biomedical applications, in particular, as load-bearing materials. However, major challenges are high stiffness ...

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the identification, localization, and characterization of nanoscale materials in biological samples is on the rise. Currently available methods, such as electron microscopy, tend to be resource-intensive, making their use prohibitive for much of the research community. Enhanced darkfield microscopy complemented with a hyperspectral imaging system may provide a solution to this bottleneck by enabling rapid and less expensive characterization of nanoparticles in histological samples. This method allows for high-contrast nanoscale imaging as well as nanomaterial identification. For this technique, histological tissue samples are prepared as they would be for light-based microscopy. First, positive control samples are analyzed to generate the reference spectra that will enable the detection of a material of interest in the sample. Negative controls without the material of interest are also analyzed in order to improve specificity (reduce false positives). Samples can then be imaged and analyzed using methods and software for hyperspectral microscopy or matched against these reference spectra in order to provide maps of the location of materials of interest in a sample. The technique is particularly well-suited for materials with highly unique reflectance spectra, such as noble metals, but is also applicable to other materials, such as semi-metallic oxides. This technique provides information that is difficult to acquire from histological samples without the use of electron microscopy techniques, which may provide higher sensitivity and resolution, but are vastly more resource-intensive and time-consuming than light microscopy.

Enhanced darkfield microscopy and hyperspectral imaging with spectral mapping enable screening, localization, and identification of nanoscale materials in histological samples with improved speed and accuracy over traditional methods. The goal of this paper is to provide methods for darkfield imaging and hyperspectral mapping of metal oxide nanoparticles in histological samples.

金属氧化物纳米颗粒的组织学样品鉴定增强暗场显微镜和高光​​谱遥感制图

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the ...

Three-Dimensionally Printed Microfluidic Cross-flow System for Ultrafiltration/Nanofiltration Membrane Performance Testing

Minimization and management of membrane fouling is a formidable challenge in diverse industrial processes and other practices that utilize membrane technology. Understanding the fouling process could lead to optimization and higher efficiency of membrane based filtration. Here we show the design and fabrication of an automated three-dimensionally (3-D) printed microfluidic cross-flow filtration system that can test up to 4 membranes in parallel. The microfluidic cells were printed using multi-material photopolymer 3-D printing technology, which used a transparent hard polymer for the microfluidic cell body and incorporated a thin rubber-like polymer layer, which prevents leakages during operation. The performance of ultrafiltration (UF), and nanofiltration (NF) membranes were tested and membrane fouling could be observed with a model foulant bovine serum albumin (BSA). Feed solutions containing BSA showed flux decline of the membrane. This protocol may be extended to measure fouling or biofouling with many other organic, inorganic or microbial containing solutions. The microfluidic design is especially advantageous for testing materials that are costly or only available in small quantities, for example polysaccharides, proteins, or lipids due to the small surface area of the membrane being tested. This modular system may also be easily expanded for high throughput testing of membranes.&#160;

Design and fabrication of a three-dimensionally (3-D) printed microfluidic cross-flow filtration system is demonstrated. The system is used to test performance and observe fouling of ultrafiltration and nanofiltration (thin film composite) membranes.

超滤/纳滤膜性能测试三维印刷微流控横流系统

Minimization and management of membrane fouling is a formidable challenge in diverse industrial processes and other practices that utilize membrane ...

Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity

Applications such as sensors, batteries, and fuel cells have been improved through the use of highly porous aerogels when functional compounds are encapsulated within the aerogels. However, few reports on encapsulating proteins within sol&#8211;gels that are processed to form aerogels exist. A procedure for encapsulating cytochrome c (cyt. c) in silica (SiO2) sol-gels that are supercritically processed to form bioaerogels with gas-phase activity for nitric oxide (NO) is presented. Cyt. c is added to a mixed silica sol under controlled protein concentration and buffer strength conditions. The sol mixture is then gelled and the liquid filling the gel pores is replaced through a series of solvent exchanges with liquid carbon dioxide. The carbon dioxide is brought to its critical point and vented off to form dry aerogels with cyt. c encapsulated inside. These bioaerogels are characterized with UV-visible spectroscopy and circular dichroism spectroscopy and can be used to detect the presence of gas-phase nitric oxide. The success of this procedure depends on regulating the cyt. c concentration and the buffer concentration and does not require other components such as metal nanoparticles. It may be possible to encapsulate other proteins using a similar approach making this procedure important for potential future bioanalytical device development.

Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity

This procedure describes how to encapsulate cytochrome c (cyt. c) in silica (SiO2) sol-gels, process these gels to form bioaerogels, and use these bioaerogels to rapidly recognize nitric oxide (NO) through a gas-phase reaction. This type of protocol may aid in the future development of biosensors or other bioanalytical devices.

细胞色素封装<em&gt;ç</em&gt;在二氧化硅气凝胶Nanoarchitectures没有金属纳米粒子，同时保持气相生物活性

Applications such as sensors, batteries, and fuel cells have been improved through the use of highly porous aerogels when functional compounds are ...

Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of medicine. In addition to efforts to identify novel small-molecule antimicrobial drugs, there has been great interest in the use of metal nanoparticles as coatings for medical devices, wound dressings, and consumer packaging, due to their antimicrobial properties. The wide variety of methods available for nanoparticle synthesis results in a broad spectrum of chemical and physical properties which can affect antibacterial efficacy. This manuscript describes the pulsed laser-ablation in liquids (PLAL) method to create nanoparticles. This approach allows for the fine tuning of nanoparticle size, composition, and stability using post-irradiation methods as well as the addition of surfactants or volume excluders. By controlling particle size and composition, a large range of physical and chemical properties of metal nanoparticles can be explored which may contribute to their antimicrobial efficacy thereby opening new avenues for antibacterial development.

The antimicrobial properties of metals such as copper and silver have been recognized for centuries. This protocol describes pulsed laser-ablation in liquids, a method of synthesizing metal nanoparticles that provides the ability to fine tune the properties of these nanoparticles to optimize their antimicrobial effects.

液体中脉冲激光烧蚀制备金属纳米粒子:研究纳米粒子抗菌性能的工具

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of ...

水样中偶极辅助固相萃取Microchip的痕量金属分析的制作

摘要

探索更多视频

此视频中的章节

无试剂检测核酸，蛋白质和小分子的电化学DNA生物传感器的制备

蛋白质和细胞坐月子创建二维图案衬底

固相Submonomer Peptoid聚合物及其自组装成高度有序的纳米片的合成

有针对性的稳定同位素稀释液相色谱 - 质谱法分析细胞脂质提取

预设计尺寸和形状的聚合物纤维和生物贿赂纤维的微流体制造

加工机械可调和生物活性金属支架的生物医学应用

金属氧化物纳米颗粒的组织学样品鉴定增强暗场显微镜和高光谱遥感制图

超滤/纳滤膜性能测试三维印刷微流控横流系统

细胞色素封装

液体中脉冲激光烧蚀制备金属纳米粒子:研究纳米粒子抗菌性能的工具

水样中偶极辅助固相萃取Microchip的痕量金属分析的制作

摘要

探索更多视频

此视频中的章节

无试剂检测核酸，蛋白质和小分子的电化学DNA生物传感器的制备

蛋白质和细胞坐月子创建二维图案衬底

固相Submonomer Peptoid聚合物及其自组装成高度有序的纳米片的合成

有针对性的稳定同位素稀释液相色谱 - 质谱法分析细胞脂质提取

预设计尺寸和形状的聚合物纤维和生物贿赂纤维的微流体制造

加工机械可调和生物活性金属支架的生物医学应用

金属氧化物纳米颗粒的组织学样品鉴定增强暗场显微镜和高光​​谱遥感制图

超滤/纳滤膜性能测试三维印刷微流控横流系统

细胞色素封装

液体中脉冲激光烧蚀制备金属纳米粒子:研究纳米粒子抗菌性能的工具

金属氧化物纳米颗粒的组织学样品鉴定增强暗场显微镜和高光谱遥感制图