我们感谢新加坡国立大学 (R-143-000-645-112、R-143-000-665-114) 和葛兰素史克-EDB (R-143-000-687-592) 提供的财政支持。

在处理任何可能的有毒和致癌化学品之前, 请参阅所有相关的材料安全数据表 (MSDS)。在开始任何反应之前进行适当的风险评估, 包括使用工程控制, 如油烟罩和气瓶, 以及佩戴足够的个人防护设备。在使用任何高度易燃气体之前, 应进行适当的训练, 以避免因气瓶处理不当而引致意外。
1. 涉及气体的反应2
<ol>
	<li>乙炔罐的研制 将乙炔罐的气体调节器设置为 20 psi (137895 pa), 高于系统中所用的 5 psi (34474 pa) 所需的背压。 
	注意:有关设置的气体调节器的详细信息, 请参阅图 1 。 
	注意:背压调节器 (BPR) 设置在油管的末端, 请参阅图 2和 3以了解有关 SFMT 设置的详细信息。</li>
	<li>4-甲醚溶液的制备
	<ol>
		<li>在10毫升的圆底烧瓶中加入一个10毫米的磁性搅拌棒。</li>
		<li>测量58.5 毫克 4-甲醚的重量平衡和转移到圆底烧瓶。 
		警告:芳香卤化物是刺激物, 可能是有害的。在继续之前, 请咨询相关 MSDSs。</li>
		<li>添加8.5 毫克 Pd (PPh3)2Cl2, 1.0mg 铜 (I) 碘化物, 21.0 毫克 1, 3, 5-苯 (内部标准) 和80µL n, n-Diisopropylethylamine (DIPEA) 到相同的圆底烧瓶中。在圆底烧瓶中加入大约2.5 毫升的二甲基亚砜。 
		警告:Pd (PPh3)2Cl2, 铜 (I) 碘化物, DIPEA 是刺激性的, 可能有害. 在继续之前, 请咨询相关 MSDSs。 
		警告: 1、3、5-苯是易燃易挥发的。远离点火源。 
		警告:亚甲基亚砜是一种有毒化学物质。在继续之前, 请咨询相关 MSDSs。</li>
		<li>用橡胶隔膜密封圆底烧瓶, 在室温和压力下将混合物搅拌在热板上, 直到所有固体都溶解。 
		注意:进一步的超声可以做, 以确保一个均匀的解决方案。</li>
		<li>将反应混合物与氩气气球混合, 在加热板上保持恒定搅拌, 约15分钟。去除两针后15分钟, 以确保一个惰性环境内的圆底烧瓶。 
		注意:有关加德过程的详细信息, 请参阅图 4 。</li>
	</ol>
	</li>
	<li>SFMT 反应器中液-气层的混合
	<ol>
		<li>从圆底烧瓶中提取所有反应混合物, 用8毫升不锈钢注射器通过橡胶隔膜通过针连接器连接到长针。取出针头并将不锈钢注射器连接到注射器泵。通过 T 型连接器将注射器连接到高纯度 Perfluoroalkoxy 烷 (HPFA) 油管 (外径 1/16 ", id 0.03", 300 厘米, 体积 = 1.37 毫升)。 
		注意:使用针接头连接不锈钢和长针, 请参阅图 5以了解有关使用针接头的详细信息。 
		注意:所有的气泡都应从不锈钢注射器中取出, 然后再附着在注射器泵上。 
		注意:确保在将反应混合物连接到设置以减少空气暴露之前, 所有油管都要拧紧, 请参阅图 2和 3连接油管。</li>
		<li>将注射器泵的流量设置为300 µL/分钟, 以便将反应混合物泵入 HPFA 油管。用针阀将乙炔的流量调整到大约1:1 液体: 沿插头的气比。平衡比是保持, 直到 HPFA 油管充满气体/液体蛞蝓试剂。 
		警告:乙炔是高度易燃的。远离点火源。 
		注意:在用乙炔气体清洗油管之前, 将 BPR 放在丙酮瓶中。 
		注意:先用乙炔气清洗油管, 直到在丙酮瓶中观察到气泡, 才能确保在将反应混合物泵入 SFMT 反应器之前, 在 SFMT 反应器内建立压力。请参阅图 6以更好地说明液体: 气体比。</li>
		<li>关闭阀门, 当所有的液体被注入到 HPFA 管或当液体开始泄漏从 BPR。泵在更多的乙炔, 直到液体停止在油管中移动, 以保持在油管内的压力。关闭阀门在起点和关闭针阀一旦完成。将整个装置转到油浴中, 孵育2小时。 
		注意:阀门保持在油浴的上方, 以防止硅油的污染。 
		注意:先将油浴加热到所需温度, 然后再将 SFMT 反应器转移到它。</li>
		<li>1小时后, 用8毫升不锈钢注射器将反应混合物泵入10毫升的小瓶中。填充8毫升不锈钢注射器与乙醚 (约4.0 毫升) 清洗任何残留的油管。 
		警告:乙醚是高度易燃的。远离火源。 
		注意:正己烷可用于清洗硅油, 然后再继续操作以避免后续步骤的污染。</li>
		<li>饱和 NH4Cl 水溶液 (4.0 毫升) 被添加到复合有机层, 其次是液-液萃取与1.5 毫升乙醚, 借助分漏斗。 
		警告:NH4Cl 可能有害。在继续之前, 请咨询相关 MSDSs。</li>
		<li>对有机层进行气相色谱质谱 (gc-ms) 分析, 以确定产量。 
		注意: 1、3、5-苯作为内部标准添加在步骤1.2.3 中。 
		注意:内部标准标定曲线是用不同质量的乘积来绘制线性回归曲线。该产品的收率从线性回归曲线中插值。有关校准曲线的详细信息, 请参阅 Ref. 2。</li>
	</ol>
	</li>
</ol>
2。照片介导的反应5
<ol>
	<li>添加30.8 毫克 benzylidenemalonitrile, 4.1 毫克 9-mesityl-10-methylacridinium 高氯酸盐, 67.3 毫克 tetramethylethylene 和2.0 毫升的氯乙烷成10毫升的硅隔膜瓶。 
	警告:Benzylidenemalonitrile, 9-mesityl-10-methylacridinium 高氯酸盐, tetramethylethylene 和氯乙烷是高度易燃。远离火源。</li>
	<li>德加约15 分钟与氩气填充气球。取出两针后15分钟, 以确保在瓶子内的惰性环境。 
	注意:有关加德过程的详细信息, 请参阅图 4 。</li>
	<li>清除 HPFA 管材 (外径 1/16 ", 身份证 0.03", 340 cm, 容量 = 1.5 毫升) 与氩气为大约5分钟由 SFMT 反应器的直接连接对氩气圆筒与一个联合身体 PEEK。关闭两个阀门, 诱骗氩气内的 HPFA 油管后达到指示时间5分钟。 
	注意:有关使用联合正文 PEEK的详细信息, 请参阅图 5 。</li>
	<li>用一个3毫升一次性注射器与长针连接, 提取反应混合物从10毫升硅隔膜瓶。取出针头, 并通过注射器连接器将一次性注射器连接到 HPFA 油管。打开两个阀门泵在反应混合物手动。再次关闭两个阀门, 一旦 HPFA 油管已填补了反应混合物。 
	注意:有关使用注射器连接器的详细信息, 请参阅图 5 。 
	注意:混合反应混合物与注射器, 以确保一个均匀的解决方案前泵入 HPFA 油管。 
	注意:可能有过剩的溶剂, 将超过油管体积。将油管末端放在废料罐上, 以收集任何溢出的反应混合物。</li>
	<li>将 SFMT 反应器置于蓝色 LED (λ最大= 425 nm、2米、20 W) 的中间, 以确保 HPFA 油管的均匀曝光。HPFA 暴露在辐射下约5-48 小时。 
	注意:蓝色 LED 条纹的长度设置为2米, 以提供足够的能量进行反应。</li>
	<li>泵出反应混合物与3毫升一次性注射器到一个干净的圆底烧瓶与一个注射器连接器片断。用3毫升一次性注射器将多余的乙醚洗净, 放入同一个圆底烧瓶。 
	注意:有关使用注射器连接器的详细信息, 请参阅图 5 。</li>
	<li>测量0.06 摩尔1、3、5-苯 (内部标准), 并加入有机混合物。用 rotavap 机在减压下去除多余的溶剂。</li>
	<li>用长针加1毫升一次性注射器测量0.6 毫升的氘氯仿, 添加到浓缩的粗品中。将氘混合物转化为一个清洁的核磁共振管, 用于原油的1H 核磁共振分析。 
	注意:6.10 ppm 的内部标准的积分 (x) 用于通过比较 3.38 ppm 形成的产品的整体 (y)来计算转换率。 
	<img alt="Equation 1" src="/files/ftp_upload/56897/56897eq1v2.jpg"></li>
</ol>
3. 照片介导的气体参与反应2
<ol>
	<li>乙炔罐的研制 将乙炔槽的气体调节器设置为约 20 psi (137895 pa), 该系统的压力高于 5 psi (34474 pa) 所需的背压。 
	注意:有关设置的气体调节器的详细信息, 请参阅图 1 。 
	注意:背压调节器 (BPR) 设置在油管的末端, 请参阅图 2 和 3以了解有关 SFMT 设置的详细信息。</li>
	<li>bromopentafluorobenzene 溶液的制备
	<ol>
		<li>在惰性气氛下, 添加74.1 毫克 bromopentafluorobenzene, 2.8 毫克 Ir (吡)2(dtbbpy) PF6和46.8 毫克 22, 66--1--1-基) 哌 (节拍) 成10毫升的硅隔膜。加入3.0 毫升的乙腈到相同的10毫升玻璃瓶溶解所有的试剂。 
		警告:Bromopentafluorobenzene 和乙腈高度易燃易挥发。远离火源。 
		警告:Ir (吡)2(dtbbpy) PF6和节拍可能有害。在继续之前, 请咨询相关 MSDSs。</li>
		<li>在冰浴中, 将反应混合物与充氩气球小心地加气10分钟。从隔膜中取出两根针, 以确保瓶子内的惰性气氛。 
		注意:有关加德过程的详细信息, 请参阅图 4 。</li>
		<li>添加56.0 µL DIPEA 到混合物与1毫升注射器和德加在一个类似于步骤3.2.2 的冰浴的另5分钟。</li>
	</ol>
	</li>
	<li>SFMT 反应器中液-气层的混合
	<ol>
		<li>用8毫升不锈钢注射器与长针连接, 通过一个针连接器, 从硅隔膜瓶中提取反应混合物。取出针头并将注射器附在注射器泵上。将插座连接至 T 型连接器。 
		注意:使用针接头连接不锈钢和长针, 请参阅图 5以了解有关使用针接头的详细信息。 
		注意:所有气体应从不锈钢注射器中取出, 然后再附着在注射器泵上。 
		注意:确保在将反应混合物连接到设置以减少气体暴露之前, 所有油管都要拧紧, 请参阅图 2和 3连接油管。</li>
		<li>将流动装置的流量设置为100 µL/分钟, 并将反应混合物泵入 HPFA 管 (外径 1/16 ", id 0.03", 300 厘米, 容量 = 1.37 毫升)。使用针阀调整乙炔的流量, 直到在插头中观察到2:1 的气液比。</li></ol></li></ol>比率插座通过估计确定在清楚的管材。 
		注意:在用乙炔气体清洗油管之前, 将 BPR 放在丙酮瓶中。 
		注意:先用乙炔气清洗油管, 直到在丙酮瓶中观察到气泡, 才能确保在将反应混合物泵入 SFMT 反应器之前, 在 SFMT 反应器内建立压力。 
		注意:请参阅图 6以更好地说明液体: 气体比, 但要注意, 气体的体积应该是通过目测的方法将液体的体积加倍。
		<li>当所有液体被注入 SFMT 反应器 (总容量0.65 毫升, 0.065 摩尔) 或当液体开始从 BPR 漏出时, 关闭阀门在末端。泵在更多的乙炔, 直到液体停止在油管中移动。关闭阀门在起点和关闭针阀一旦完成。将整个设置转换为水浴预热至60° c, 并允许在蓝色 LED 灯下 (λmax = 425 nm, 3 米, 30 W) 下对 3 h 进行反应。 
		注意:阀门留在水浴上方以防止任何污染。 
		注意:蓝色 LED 条纹的长度设置为3米, 以提供足够的能量进行反应。</li>
		<li>泵的反应混合物从 HPFA 油管与8毫升不锈钢注射器到一个圆底烧瓶。用过量的乙醚将管式反应器中的残留物冲洗到同一个圆底烧瓶中。用 rotavap 机将混合物集中在减压的压力下。 
		注意:小心地减少压力, 因为开始的材料和产品是高度挥发性的。</li>
		<li>将0.6 毫升的氘氯仿通过1毫升一次性注射器放入圆底烧瓶中, 以溶解浓缩的原油混合物。将氘混合物转化为核磁共振导管, 用于19F 核磁共振分析。 
		注意:对起始材料 (bromopentafluorobenzene) 和2产品 (2、3、4、5、6-Pentafluorostyrene 和氟) 的19F 核磁共振谱进行了分析, 找出了每个化学物质的一个重要峰值。粗略的19F 核磁共振谱用于比较这3个重要峰值的积分, 以确定产品形成的比率。有关产品转换和产品比率计算的详细信息, 请参阅 Ref. 2。</li>

<ol><li>Mallia, C. J., Baxendale, I. R. <a target="_blank" href="http://pubs.acs.org/doi/abs/10.1021/acs.oprd.5b00222">The Use of Gases in Flow Synthesis.</a> Organic Process Research & Development. 20 (2), 327-360 (2016).</li>
<li>Xue, F., Deng, H., Xue, C., Mohamed, D. K. B., Tang, K. Y., Wu, J. <a target="_blank" href="http://pubs.rsc.org/en/content/articlelanding/2017/sc/c7sc00408g#!divAbstract">Reaction discovery using acetylene gas as the chemical feedstock accelerated by the &#34;stop-flow&#34; micro-tubing reactor system.</a> Chemical Science. 8 (5), 3623-3627 (2017).</li>
<li>McTeague, T. A., Jamison, T. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Photoredox+Activation+of+SF6+for+Fluorination%5BTitle%5D)+AND+%22Angewandte+Chemie+International+Edition%22%5BJournal%5D)">Photoredox Activation of SF6 for Fluorination.</a> Angewandte Chemie International Edition. 55 (48), 15072-15075 (2016).</li>
<li>Mohamed, D. K. B., Yu, X., Li, J., Wu, J. <a target="_blank" href="http://www.sciencedirect.com/science/article/pii/S0040403916309297">Reaction screening in continuous flow reactors.</a> Tetrahedron Letters. 57 (36), 3965-3977 (2016).</li>
<li>Zhou, R., Liu, H., Tao, H., Yu, X., Wu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Metal-free+direct+alkylation+of+unfunctionalized+allylic%2Fbenzylic+sp3+C-H+bonds+via+photoredox+induced+radical+cation+deprotonation%5BTitle%5D)+AND+%22Chemical+Science%22%5BJournal%5D)">Metal-free direct alkylation of unfunctionalized allylic/benzylic sp3 C-H bonds via photoredox induced radical cation deprotonation.</a> Chemical Science. 8 (6), 4654-4659 (2017).</li>
<li>Prier, C. K., Rankic, D. A., MacMillan, D. W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Visible+Light+Photoredox+Catalysis+with+Transition+Metal+Complexes%3A+Applications+in+Organic+Synthesis%5BTitle%5D)+AND+%22Chemical+Reviews%22%5BJournal%5D)">Visible Light Photoredox Catalysis with Transition Metal Complexes: Applications in Organic Synthesis.</a> Chemical Reviews. 113 (7), 5322-5363 (2013).</li>
<li>Cambié, D., Bottecchia, C., Straathof, N. J. W., Hessel, V., Noël, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Applications+of+Continuous-Flow+Photochemistry+in+Organic+Synthesis%2C+Material+Science%2C+and+Water+Treatment%5BTitle%5D)+AND+%22Chemical+Reviews%22%5BJournal%5D)">Applications of Continuous-Flow Photochemistry in Organic Synthesis, Material Science, and Water Treatment.</a> Chemical Reviews. 116 (17), 10276-10341 (2016).</li>
<li>Straathof, N. J. W., Su, Y., Hessel, V., Noel, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Accelerated+gas-liquid+visible+light+photoredox+catalysis+with+continuous-flow+photochemical+microreactors%5BTitle%5D)+AND+%22Nat.+Protocols%22%5BJournal%5D)">Accelerated gas-liquid visible light photoredox catalysis with continuous-flow photochemical microreactors.</a> Nat. Protocols. 11 (1), 10-21 (2016).</li>
<li>Robards, K., Haddad, P. R., Jackson, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((High-performance+Liquid+Chromatography%E2%80%94Instrumentation+and+Techniques%5BTitle%5D)+AND+%22Principles+and+Practice+of+Modern+Chromatographic+Methods%22%5BJournal%5D)">High-performance Liquid Chromatography—Instrumentation and Techniques.</a> Principles and Practice of Modern Chromatographic Methods. 5, 227-303 (1994).</li>
<li>Linder, V., Sia, S. K., Whitesides, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reagent-Loaded+Cartridges+for+Valveless+and+Automated+Fluid+Delivery+in+Microfluidic+Devices%5BTitle%5D)+AND+%22Analytical+Chemistry%22%5BJournal%5D)">Reagent-Loaded Cartridges for Valveless and Automated Fluid Delivery in Microfluidic Devices.</a> Analytical Chemistry. 77 (1), 64-71 (2005).</li>
<li>Terao, K., Nishiyama, Y., Tanimoto, H., Morimoto, T., Oelgemöller, M., Morimoto, T. <a target="_blank" href="http://akademiai.com/doi/abs/10.1556/JFC-D-12-00005">Diastereoselective [2+2] Photocycloaddition of a Chiral Cyclohexenone with Ethylene in a Continuous Flow Microcapillary Reactor.</a> Journal of Flow Chemistry. 2 (3), 73-76 (2012).</li>
<li>Qian, D., Lawal, A. <a target="_blank" href="http://www.sciencedirect.com/science/article/pii/S0009250906005604">Numerical study on gas and liquid slugs for Taylor flow in a T-junction microchannel.</a> Chemical Engineering Science. 61 (23), 7609-7625 (2006).</li>
<li>Hamilton, D. S., Nicewicz, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Direct+Catalytic+Anti-Markovnikov+Hydroetherification+of+Alkenols%5BTitle%5D)+AND+%22Journal+of+the+American+Chemical+Society%22%5BJournal%5D)">Direct Catalytic Anti-Markovnikov Hydroetherification of Alkenols.</a> Journal of the American Chemical Society. 134 (45), 18577-18580 (2012).</li>
<li>Singh, A., Teegardin, K., Kelly, M., Prasad, K. S., Krishnan, S., Weaver, J. D. <a target="_blank" href="http://www.sciencedirect.com/science/article/pii/S0022328X14005208">Facile synthesis and complete characterization of homoleptic and heteroleptic cyclometalated Iridium(III) complexes for photocatalysis.</a> Journal of Organometallic Chemistry. 776, 51-59 (2015).</li>
</ol>

作者没有什么可透露的。

新开发的 SFMT 反应器是通过在微导管2上添加关闭阀来改进连续流系统。在这个系统中, 所需的试剂量的流速可以停止, 模拟一个间歇反应堆, 但在微管2,10,11。这些阀门有助于在 HPFA 或不锈钢管中捕获所需的试剂量, 同时保持内部压力, 类似于高压容器。这种方便的系统能够通过同时设置...

流动化学对绿色和可持续过程的运动有很好的准备1,2。与间歇式反应器相比, 连续流反应器具有显著的优点, 如改进的热管理、强化的混合控制和内部压力调节。这些优点大大减少了连续流系统中 by-products 的形成。此外, 连续流增强了微管内的双相气液反应, 由于不同状态的试剂界面表面积很好。连续流反应器也提供了一个良好的光合作用的平台, 由于增强和统一的光照明横跨微型油管3。
尽管在连续流技术方面取得了成功, 但在涉及催化剂、溶剂和试剂的反应筛选方面仍有局限性2。对流量系统压力的变化将会极大地影响水流平衡。此外, 一个经典的连续流系统一般只限于一次反应筛选, 使它的时间消耗有效的平行反应筛选。连续流合成的反应时间也受其微管反应器大小的限制。此外, 即使在不同的反应之间使用载体介质4, 连续流筛选也容易在较高温度下进行交叉污染。
因此, 为了解决在连续流系统中筛选离散参数的困难, 我们开发了一种用于反应筛查的停止流动微管 (SFMT) 反应器系统, 它涉及气态试剂和/或光介导的反应2。SFMT 反应堆包括间歇反应堆和连续流动反应器的元素。关闭阀的引入 entraps 了微型油管内的试剂, 这一概念与间歇式反应器类似, 当系统被加压时, SFMT 的作用是微型高压反应器。然后, SFMT 可以被淹没到水或油浴中, 将热量引入反应堆系统。在反应期间, 可见光也可以在微管上发光, 以促进照片介导的反应。
在 SFMT, 可燃或有毒气体, 如乙烯、乙炔和一氧化碳, 可用于生成有价值的化学物质的安全方式相比, 批处理反应器1,2,4。它是一种资产, 使用这种反应性气体, 因为它们是廉价的化学原料, 可以很容易地消除后, 反应完成, 提供一个更清洁的程序2。相反, 在间歇式反应堆中进行的大多数反应发展往往不使用反应性气体, 因为它的不便和爆炸的风险在升高的压力和温度。如果使用气态试剂, 通常通过冒泡或气球将它们引入间歇反应器。由于界面的混合效率低, 这通常会降低重复性或反应性。虽然高压容器通常用于提高气体的反应性和溶解度, 但它们是很费力的, 有爆炸的危险, 特别是易燃气体。此外, 那些通常使用的高压反应堆的不透明表面使它不适合于光介导的反应。因此, 由气态试剂和光介导的反应组成的反应一般未被探索。在这种情况下, SFMT 反应堆提供了一个理想的平台, 因为气体试剂可以在微导管内使用, 在背压调节器 (BPR) 的协助下, 以安全和方便的方式调节内部压力2。除涉及气态试剂的反应外, 可见光促进合成还显示了有机合成的巨大承诺5,6。然而, 可见光介导的反应的最大的衰落之一是传统间歇式反应器的伸缩性, 因为在大型容器中光子传输的衰减效应7。如果使用大功率光源, over-irradiation 可能导致 by-product 形成。此外, 气态试剂很少被应用于光化学反应, 主要是由于复杂的仪器系统使用气相反应物在高压2。通过引入窄通道, 如 SFMT, 可以在光照射下轻松实现高压气体环境。
因此, 这一详细的视频旨在帮助更多的科学家了解使用 SFMT 进行气体转化和 light-mediated 反应的条件筛选的优点和程序。

<table><tbody><tr><td>Acetylene Cylinder</td><td>Chem Gas PTE LTD (Singapore)</td><td></td><td></td></tr><tr><td>Logato 200 series Syringe pumps</td><td>KD Scientific Inc</td><td>788200</td><td></td></tr><tr><td>Blue LED Strips</td><td>Inwares Pte Ltd (Singapore)</td><td>3528 FlexiGlow LED Strips</td><td></td></tr><tr><td>PFA Tubing High Purity 1/16&quot; OD x .030&quot; ID x 50ft</td><td>IDEX Health&amp;amp;Science</td><td>1632-L</td><td>Depending on diameter of tubings needed</td></tr><tr><td>KDS Stainless Steel Syringe</td><td>KD Scientific Inc</td><td>780802</td><td></td></tr><tr><td>Shut-Off Valve Tefzel (ETFE) with 1/16&quot; Fittings</td><td>IDEX Health&amp;amp;Science</td><td>P-782</td><td></td></tr><tr><td>BPR Assembly 20 psi</td><td>IDEX Health&amp;amp;Science</td><td>P-791</td><td></td></tr><tr><td>Luer Adapter Female Luer - Female Union</td><td>IDEX Health&amp;amp;Science</td><td>P-628</td><td>Known as syringe connector in this paper</td></tr><tr><td>1/4-28 Female to Male Luer Assy</td><td>IDEX Health&amp;amp;Science</td><td>P-675</td><td>Known as needle connector in this paper</td></tr><tr><td>Union Body PEEK .020 thru hole, for 1/16&quot; OD&quot;</td><td>IDEX Health&amp;amp;Science</td><td>P-702-01</td><td></td></tr><tr><td>Super Flangeless Ferrule w/SST Ring, 1/4-28 Flat-Bottom, for 1/16&quot; OD</td><td>IDEX Health&amp;amp;Science</td><td>P-250X</td><td></td></tr><tr><td>PEEK Low Pressure Tee Assembly 1/16&quot; PEEK .020 thru hole</td><td>IDEX Health&amp;amp;Science</td><td>P-712</td><td>Known as T-connector in this paper</td></tr><tr><td>Super Flangeless Nut PEEK 1/4-28 Flat-Bottom, for 1/16&quot; &amp;amp; 1/32&quot; OD</td><td>IDEX Health&amp;amp;Science</td><td>P-255X</td><td></td></tr><tr><td>Micro Metering Valve Assembly, 1/4-28 Flat-Bottom, for 1/16&quot; OD</td><td>IDEX Health&amp;amp;Science</td><td>P-445NF</td><td>Known as Needle valve in this paper</td></tr><tr><td>Shut Off Valve Assembly PEEK .020</td><td>IDEX Health&amp;amp;Science</td><td>P-732</td><td></td></tr><tr><td>Terumo Syringe without needle</td><td>Terumo medical</td><td></td><td>1 mL and 3 mL depending on the volume needed</td></tr><tr><td>Terumo needle</td><td>Terumo medical</td><td></td><td>22G X 1&amp;frac12;&amp;rdquo;&lt;br/&gt; (0.70 X 38 mm)</td></tr><tr><td>Sterican needle</td><td>B | Braun Sharing Enterprise</td><td></td><td>21G X 4&amp;frac34;&amp;rdquo;&lt;br/&gt; (0.80 X 120 mm)</td></tr><tr><td>Bruker ACF300 (300 MHz)</td><td></td><td></td><td>For 300 MHz NMR scanning</td></tr><tr><td>AV-III400 (400 MHZ)</td><td></td><td></td><td>For 400 MHz NMR scanning</td></tr><tr><td>AMX500 (500 MHz)</td><td></td><td></td><td>For 500 MHz NMR scanning</td></tr><tr><td>Merck 60 (0.040-0.063 mm) mesh silica gel</td><td>Merck</td><td></td><td></td></tr><tr><td>4-Iodoanisole</td><td>Sigma Aldrich</td><td>I7608-100G</td><td></td></tr><tr><td>412740 ALDRICH&lt;br/&gt; Bis(triphenylphosphine)&lt;br/&gt; palladium(II) dichloride&lt;br/&gt; &amp;ge;99% trace metals basis</td><td>Sigma Aldrich</td><td>412740-5G</td><td></td></tr><tr><td>Copper(I) iodide&lt;br/&gt; purum, &amp;ge;99.5%</td><td>Sigma Aldrich</td><td>03140-100G</td><td></td></tr><tr><td>N,N-Diisopropylethylamine</td><td>Tokyo Chemical Industry Co., Ltd</td><td>D1599</td><td></td></tr><tr><td>1, 3, 5-trimethoxybenzene</td><td>Tokyo Chemical Industry Co., Ltd</td><td>P0250</td><td></td></tr><tr><td>2,3-Dimethyl-2-butene&lt;br/&gt; &amp;ge;99%</td><td>Sigma Aldrich</td><td>220159-25ML</td><td></td></tr><tr><td>Bromopentafluorobenzene&lt;br/&gt; 99%</td><td>Sigma Aldrich</td><td>B75158-10G</td><td></td></tr><tr><td>TEMPO Green Alternative&lt;br/&gt; 98%</td><td>Sigma Aldrich</td><td>214000-25G</td><td></td></tr><tr><td>Acetonitrile</td><td>Sigma Aldrich</td><td>271004-1L</td><td></td></tr><tr><td>Diethylether</td><td>Sigma Aldrich</td><td>346136-1L</td><td></td></tr><tr><td>Dimethyl sulfoxide</td><td>VWR chemical</td><td>23500.322- 25L</td><td></td></tr><tr><td>1,2-Dichloroethane</td><td>Sigma Aldrich</td><td>284505-1L</td><td></td></tr><tr><td>9-mesityl-10-methylacridinium perchlorate</td><td></td><td></td><td>Refer to Ref. &lt;sup class=&quot;xref&quot;&gt;8&lt;/sup&gt; for synthesis</td></tr><tr><td>Ir(ppy)&lt;sub&gt;2&lt;/sub&gt;(dtbbpy)PF&lt;sub&gt;6&lt;/sub&gt;</td><td></td><td></td><td>Refer to Ref. &lt;sup class=&quot;xref&quot;&gt;9&lt;/sup&gt; for synthesis</td></tr></tbody></table>

utilization of stop-flow micro-tubing reactors for the development of organic transformations

最近, 通过将连续微流和常规间歇反应器中的元素相结合, 产生了一种新的有机合成反应筛选技术, 从而生成了停止流动微管 (SFMT) 反应器。在 SFMT, 需要高压的化学反应可以通过更安全和方便的方式进行平行筛选。在连续流反应器的反应筛查中, 交叉污染是 SFMT 的一个常见问题。此外, 商业可用的 light-permeable 微管可以并入 SFMT, 作为一个极好的选择 light-mediated 反应, 由于更有效的均匀光曝光, 与间歇式反应堆。总的来说, SFMT 反应堆系统类似于连续流反应器和比间歇反应器更优越的反应, 包括气体试剂和/或要求 light-illumination, 这使得一个简单但高效的反应筛选系统。此外, 在 SFMT 反应器系统中任何成功发展的反应都可以方便地转化为大规模生产的连续流合成。

在这项研究中, SFMT 被用来进行转化, 包括气态试剂 (表 1), light-mediated 反应 (表 2), 以及涉及气体试剂和光催化反应 (表 3)。
图 1显示了一个典型的设置, 用于将气体调节器连接到气瓶, 以便调节被泵入 SFMT 系统的气体的压力。
图 2代表了如何将试剂连接到 SFMT 系统中的试剂的设置。油管的长度取决于反应中所使用...

Watch this Scientific Journal Video about Utilization of Stop-flow Micro-tubing Reactors for the Development of Organic Transformations at JoVE.com

Utilization of Stop-flow Micro-tubing Reactors for the Development of Organic Transformations

本文介绍了利用气态反应物和/或可见光介导的反应, 采用停止流动微管 (SFMT) 反应器进行有机反应筛选的协议。

停止流动微管反应器在有机转化发展中的应用

A new reaction screening technology for organic synthesis was recently demonstrated by combining elements from both continuous micro-flow and conventional ...

utilization-stop-flow-micro-tubing-reactors-for-development-organic

Research

JoVE Journal

Chemistry

39.1K 次观看.  National University of Singapore. 本文介绍了利用气态反应物和/或可见光介导的反应, 采用停止流动微管 (SFMT) 反应器进行有机反应筛选的协议。

视频: Utilization of Stop-flow Micro-tubing Reactors for the Development of Organic Transformations

Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example

By using inline monitoring, it is possible to optimize reactions performed using continuous-flow processing in a simple and rapid way. It is also possible to ensure consistent product quality over time using this technique. We here show how to interface a commercially available flow unit with a Raman spectrometer. The Raman flow cell is placed after the back-pressure regulator, meaning that it can be operated at atmospheric pressure. In addition, the fact that the product stream passes through a length of tubing before entering the flow cell means that the material is at RT. It is important that the spectra are acquired under isothermal conditions since Raman signal intensity is temperature dependent. Having assembled the apparatus, we then show how to monitor a chemical reaction, the piperidine-catalyzed synthesis of 3-acetylcoumarin from salicylaldehyde and ethyl acetoacetate being used as an example. The reaction can be performed over a range of flow rates and temperatures, the in-situ monitoring tool being used to optimize conditions simply and easily.

Real-time monitoring allows for fast optimization of reactions performed using continuous-flow processing. Here the preparation of 3-acetylcoumarin is used as an example. The apparatus for performing in-situ Raman monitoring is described, as are the steps required to optimize the reaction.

反应执行使用连续流处理的实时监控:3  - 乙酰基香豆素的制备为例

By using inline monitoring, it is possible to optimize reactions performed using continuous-flow processing in a simple and rapid way. It is also possible ...

科研

化学

Production and Measurement of Organic Particulate Matter in a Flow Tube Reactor

Organic particulate matter (PM) is increasingly recognized as important to the Earth's climate system as well as public health in urban regions, and the production of synthetic PM for laboratory studies have become a widespread necessity. Herein, experimental protocols demonstrate approaches to produce aerosolized organic PM by &#945;-pinene ozonolysis in a flow tube reactor. Methods are described for measuring the size distributions and morphology of the aerosol particles. The video demonstrates basic operations of the flow tube reactor and related instrumentation. The first part of the video shows the procedure for preparing gas-phase reactants, ozonolysis, and production of organic PM. The second part of the video shows the procedures for determining the properties of the produced particle population. The particle number-diameter distributions show different stages of particle growth, namely condensation, coagulation, or a combination of both, depending on reaction conditions. The particle morphology is characterized by an aerosol particle mass analyzer (APM) and a scanning electron microscope (SEM). The results confirm the existence of non-spherical particles that have grown from coagulation for specific reaction conditions. The experimental results also indicate that the flow tube reactor can be used to study the physical and chemical properties of organic PM for relatively high concentrations and short time frames.

This paper describes the operation procedure for the flow tube reactor and related data collection. It shows the protocols for setting the experiments, recording data and generating the number-diameter distribution as well as the particle mass information, which gives useful information about chemical and physical properties of the organic aerosols.

流管反应器中有机颗粒物的生产与测定

Organic particulate matter (PM) is increasingly recognized as important to the Earth's climate system as well as public health in urban regions, and the ...

环境科学

Microfluidic Picoliter Bioreactor for Microbial Single-cell Analysis: Fabrication, System Setup, and Operation

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described in detail. The PLBR can be utilized for the analysis of single bacteria and microcolonies to investigate biotechnological and microbiological related questions concerning, e.g.&#160;cell growth, morphology, stress response, and metabolite or protein production on single-cell level. The device features continuous media flow enabling constant environmental conditions for perturbation studies, but in addition allows fast medium changes as well as oscillating conditions to mimic any desired environmental situation. To fabricate the single use devices, a silicon wafer containing sub micrometer sized SU-8 structures served as the replication mold for rapid polydimethylsiloxane casting. Chips were cut, assembled, connected, and set up onto a high resolution and fully automated microscope suited for time-lapse imaging, a powerful tool for spatio-temporal cell analysis. Here, the biotechnological platform organism Corynebacterium glutamicum&#160;was seeded into the PLBR and cell growth and intracellular fluorescence were followed over several hours unraveling time dependent population heterogeneity on single-cell level, not possible with conventional analysis methods such as flow cytometry. Besides insights into device fabrication, furthermore, the preparation of the preculture, loading, trapping of bacteria, and the PLBR cultivation of single cells and colonies is demonstrated. These devices will add a new dimension in microbiological research to analyze time dependent phenomena of single bacteria under tight environmental control. Due to the simple and relatively short fabrication process the technology can be easily adapted at any microfluidics lab and simply tailored towards specific needs.

In this protocol the fabrication, setup and basic operation of a microfluidic picoliter bioreactor (PLBR) for single-cell analysis of prokaryotic microorganisms is introduced. Industrially relevant microorganisms were analyzed as proof of principle allowing insights into growth rate, morphology, and phenotypic heterogeneity over certain time periods, hardly possible with conventional methods.

In this protocol the fabrication, experimental setup and basic operation of the recently introduced microfluidic picoliter bioreactor (PLBR) is described ...

生物工程

Millifluidics for Chemical Synthesis and Time-resolved Mechanistic Studies

Procedures utilizing millifluidic devices for chemical synthesis and time-resolved mechanistic studies are described by taking three examples. In the first, synthesis of ultra-small copper nanoclusters is described. The second example provides their utility for investigating time resolved kinetics of chemical reactions by analyzing gold nanoparticle formation using in situ X-ray absorption spectroscopy. The final example demonstrates continuous flow catalysis of reactions inside millifluidic channel coated with nanostructured catalyst.

Millifluidic devices are utilized for controlled synthesis of nanomaterials, time-resolved analysis of reaction mechanisms and continuous flow catalysis.

Millifluidics的化学合成和时间分辨机理研究

Procedures utilizing millifluidic devices for chemical synthesis and time-resolved mechanistic studies are described by taking three examples. In the ...

Continuous Flow Chemistry: Reaction of Diphenyldiazomethane with p-Nitrobenzoic Acid

Continuous flow technology has been identified as instrumental for its environmental and economic advantages leveraging superior mixing, heat transfer and cost savings through the &#34;scaling out&#34; strategy as opposed to the traditional &#34;scaling up&#34;. Herein, we report the reaction of diphenyldiazomethane with p-nitrobenzoic acid in both batch and flow modes. To effectively transfer the reaction from batch to flow mode, it is essential to first conduct the reaction in batch. As a consequence, the reaction of diphenyldiazomethane was first studied in batch as a function of temperature, reaction time, and concentration to obtain kinetic information and process parameters. The glass flow reactor set-up is described and combines two types of reaction modules with &#34;mixing&#34; and &#34;linear&#34; microstructures. Finally, the reaction of diphenyldiazomethane with p-nitrobenzoic acid was successfully conducted in the flow reactor, with up to 95% conversion of the diphenyldiazomethane in 11 min. This proof of concept reaction aims to provide insight for scientists to consider flow technology&#39;s competitiveness, sustainability, and versatility in their research.

Continuous Flow Chemistry: Reaction of Diphenyldiazomethane with p-Nitrobenzoic Acid

Flow chemistry carries environmental and economic advantages by leveraging superior mixing, heat transfer and cost benefits. Herein, we provide a blueprint to transfer chemical processes from batch to flow mode. The reaction of diphenyldiazomethane (DDM) with p-nitrobenzoic acid, conducted in batch and flow, was chosen for proof of concept.

连续流动化学: Diphenyldiazomethane 与p硝基苯甲酸的反应

Continuous flow technology has been identified as instrumental for its environmental and economic advantages leveraging superior mixing, heat transfer and ...

A Scalable Balz-Schiemann Reaction Protocol in a Continuous Flow Reactor

The demand for aromatic fluorides is steadily increasing in the pharmaceutical and fine chemical industries. The Balz-Schiemann reaction is a straightforward strategy for preparing aryl fluorides from aryl amines, via the preparation and conversion of diazonium tetrafluoroborate intermediates. However, significant safety risks exist in handling the aryl diazonium salts when scaling up. In order to minimize the hazard, we present a continuous flow protocol that has been successfully performed at a kilogram scale that eliminates the isolation of aryl diazonium salts while facilitating efficient fluorination. The diazotization process was performed at 10 &#176;C with a residence time of 10 min, followed by a fluorination process at 60 &#176;C with a residence time of 5.4 s with about 70% yield. The reaction time has been dramatically reduced by introducing this multi-step continuous flow system.

A detailed scalable continuous flow protocol is presented to synthesize an aryl fluoride from an aryl amine through the Balz-Schiemann reaction.

连续流反应器中的可扩展巴尔茨-席曼反应方案

The demand for aromatic fluorides is steadily increasing in the pharmaceutical and fine chemical industries. The Balz-Schiemann reaction is a ...

Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of bioprocess conditions. One important aspect is the improvement of cultivation medium to provide an optimal environment for microbial formation of the product of interest. It is well accepted that the media composition can dramatically influence overall bioprocess performance. Nutrition medium optimization is known to improve recombinant protein production with microbial systems and thus, this is a rewarding step in bioprocess development. However, very often standard media recipes are taken from literature, since tailor-made design of the cultivation medium is a tedious task that demands microbioreactor technology for sufficient cultivation throughput, fast product analytics, as well as support by lab robotics to enable reliability in liquid handling steps. Furthermore, advanced mathematical methods are required for rationally analyzing measurement data and efficiently designing parallel experiments such as to achieve optimal information content.
The generic nature of the presented protocol allows for easy adaption to different lab equipment, other expression hosts, and target proteins of interest, as well as further bioprocess parameters. Moreover, other optimization objectives like protein production rate, specific yield, or product quality can be chosen to fit the scope of other optimization studies. The applied Kriging Toolbox (KriKit) is a general tool for Design of Experiments (DOE) that contributes to improved holistic bioprocess optimization. It also supports multi-objective optimization which can be important in optimizing both upstream and downstream processes.

This manuscript describes a generic approach for tailor-made design of microbial cultivation media. This is enabled by an iterative workflow combining Kriging-based experimental design and microbioreactor technology for sufficient cultivation throughput, which is supported by lab robotics to increase reliability and speed in liquid handling media preparation.

利用自动 Microbioreactor 技术优化异种蛋白质生产的通用协议

A core business in industrial biotechnology using microbial production cell factories is the iterative process of strain engineering and optimization of ...

Use of High-Throughput Automated Microbioreactor System for Production of Model IgG1 in CHO Cells

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of experimental conditions while minimizing potential process variability. Applications of this approach include: clone screening, temperature and pH shifts,&#160;media and supplement optimization. Furthermore, the small reactor volumes are conducive to large Design of Experiments that investigate a wide range of conditions. This allows upstream processes to be significantly optimized before scale-up where experimentation is more limited in scope due to time and economic constraints. Automated microscale bioreactor systems offer various advantages over traditional small scale cell culture units, such as shake flasks or spinner flasks. However, during pilot scale process development&#160;significant care must be taken to ensure that these advantages are realized. When run with care, the system can enable high level automation, can be programmed to run DOE&#39;s with a higher number of variables&#160;and can reduce sampling time when integrated with a nutrient analyzer or cell counter. Integration of the expert-derived heuristics presented here, with current automated microscale bioreactor experiments can minimize common pitfalls that hinder meaningful results. In the extreme, failure to adhere to the principles laid out here can lead to equipment damage that requires expensive repairs. Furthermore, the microbioreactor systems have small culture volumes&#160;making characterization of cell culture conditions difficult. The number and amount of samples taken in-process in batch mode culture is limited as operating volumes cannot fall below 10 mL. This method will discuss the benefits and drawbacks of microscale bioreactor systems.

A detailed protocol for the concurrent operation of 48 parallel cell cultures under varied conditions in a microbioreactor system is presented. Cell culture process, harvest and subsequent antibody titer analysis are described.

高通量自动 Microbioreactor 系统在 CHO 细胞模型 IgG1 生产中的应用

Automated microscale bioreactors (15 mL) can be a useful tool for cell culture engineers. They facilitate the simultaneous execution of a wide variety of ...

A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression

The limitations of cell-based synthetic biology are becoming increasingly apparent as researchers aim to develop larger and more complex synthetic genetic regulatory circuits. The analysis of synthetic genetic regulatory networks in vivo is time consuming and suffers from a lack of environmental control, with exogenous synthetic components interacting with host processes resulting in undesired behavior. To overcome these issues, cell-free characterization of novel circuitry is becoming more prevalent. In vitro transcription and translation (IVTT) mixtures allow the regulation of the experimental environment and can be optimized for each unique system. The protocols presented here detail the fabrication of a multilayer microfluidic device that can be utilized to sustain IVTT reactions for prolonged durations. In contrast to batch reactions, where resources are depleted over time and (by-) products accumulate, the use of microfluidic devices allows the replenishment of resources as well as the removal of reaction products. In this manner, the cellular environment is emulated by maintaining an out-of-equilibrium environment in which the dynamic behavior of gene circuits can be investigated over extended periods of time. To fully exploit the multilayer microfluidic device, hardware and software have been integrated to automate the IVTT reactions. By combining IVTT reactions with the microfluidic platform presented here, it becomes possible to comprehensively analyze complex network behaviors, furthering our understanding of the mechanisms that regulate cellular processes.

The fabrication process of a PDMS-based, multilayer, microfluidic device that allows in vitro transcription and translation (IVTT) reactions to be performed over prolonged periods is described. Furthermore, a comprehensive overview of the hardware and software required to automate and maintain these reactions for prolonged durations is provided.

用于长期无细胞基因表达传导的多层微流体平台

The limitations of cell-based synthetic biology are becoming increasingly apparent as researchers aim to develop larger and more complex synthetic genetic ...

Process Optimization using High Throughput Automated Micro-Bioreactors in Chinese Hamster Ovary Cell Cultivation

Optimization of bioprocesses to increase the yield of desired products is of importance in the biopharmaceutical industry. This can be achieved by strain selection and by developing bioprocess parameters. Shake flasks have been used for this purpose. They, however, lack the capability to control the process parameters such as pH and dissolved oxygen (DO). This limitation can be overcome with the help of an automated micro-bioreactor. These bioreactors mimic cultivation at a larger scale. One of the major advantages of this system is the integration of the Design of Experiment (DOE) in the software. This integration enables establishing a design where multiple process parameters can be varied simultaneously. The critical process parameters and optimum bioprocess conditions can be analyzed within the software. The focus of the work presented here is to introduce the user to the steps involved in process design in the software and incorporation of the DOE within the cultivation run.

Here, we present a detailed procedure to run a Design of Experiment in an automated micro-bioreactor followed by cell harvest and protein quantification using a Protein A column.

中国仓鼠卵巢细胞培养中高通量自动化微生物反应器工艺优化

Optimization of bioprocesses to increase the yield of desired products is of importance in the biopharmaceutical industry. This can be achieved by strain ...