Financial support provided by National Science Foundation (CAREER award CHE-0847262. We thank Vapourtec Ltd and Enwave Optronics for equipment support, and Daniel Daleb of the University of Connecticut for his assistance in construction of the flow cell apparatus.

1. Reaksiyon İzleme uygun Sinyalleri bulun <ol><li> Tüm başlangıç ​​malzemeleri ve ürün için Raman edinin. </li><li> Spektrumları Yerleşimi ve ürüne özgü yoğun bir bant tespit. </li><li> Reaksiyonun ilerlemesini izlemek için bu Raman bandı kullanın. 1608 cm &#39;de bir bant -1 bu durumda seçildi. </li></ol> 2. Akış Hücre kurun <ol><li> Uygun bir akış hücresi elde edilir. 6,5 mm genişlik, 20 mm yükseklik ve 5 mm (Şekil 2A) &#39;in bir yol uzunluğu: Burada, aşağıdaki boyutlara sahip bir kullanın. </li><li> Ortam ışığının serbest bir ortam sağlar bir kapta akış hücresi yerleştirin. </li><li> (Bu durumda 1 mm ID PFA tüp) akış hücresi giriş ve çıkışına hortumu bağlayın. </li></ol> 3. Akış Hücre ile Raman Spektrometresi Arabirimi <ol><li> C yerleştirilebilir esnek bir optik tertibatı ile uygun bir Raman spektrometresi Eldeakış hücresi yakınlığı kaybedersiniz. </li><li> Akış hücresi düzeneğinin (Şekil 2B) ihtiva eden kutuya uygun boyutlu bir açıklık boyunca optik tertibatım yerleştirin. </li><li> Akış hücresi dokunana kadar optik aksamını kaydırın ve daha sonra ~ 2 mm&#39;lik bir boşluk bırakarak geri çekin. </li><li> % 100 aseton ile akış hücresi doldurun. </li><li> Raman spektrometresi açın ve sürekli tarama modunda spektrumları kazanır. </li><li> Hafifçe bir seferde ışık borusunun bir kısmını hareket ettirerek lazer odaklanın. Sinyal onun en büyük yoğunlukta ve zirveleri keskin ve iyi tanımlanmış kadar hafif boru hareketli tutun. </li></ol><img alt="Şekil 2," src="/files/ftp_upload/52393/52393fig2.jpg" /> Şekil 2. (A) hücre ve kullanılan (B) Raman arayüzünü Akış. <a href="https://www.jove.com/files/ftp_upload/52393/52393fig2large.jpg" target="_blank">Daha büyük bir ettik görmek için buraya tıklayınız</a>Bu rakamın rsion. 4. Reaktif ve Solvent Çözümler hazırlayın <ol><li> 50 ml&#39;lik volümetrik şişeye (6,106, 50 mmol gr, 1 eşdeğer) ve etil asetoasetat (6,507 g, 50 mmol, 1 eşdeğer) salisilaldehit ekleyin. </li><li> 50 ml&#39;lik bir toplam hacme etil asetat ekleyin ve daha sonra içeriğinin iyice karıştırın. </li><li> Bir manyetik karıştırma çubuğu içeren 20 ml&#39;lik bir cam şişeye stok çözeltisi, 10 ml&#39;lik bir şişe aktarın. Bu flakon &quot;ayıracı.&quot; Etiket </li><li> 100 ml&#39;lik bir şişe, etil asetat yerine 90 ml içinde. &quot;Solvent&quot; bu şişeyi etiketleyin. 100 ml&#39;lik bir şişe yer aseton 90 ml içinde. Bu şişeyi &quot;solvent yolunu kesmek&quot; etiketleyin. </li></ol> 5. Akış Aparatı hazırlayın <ol><li> &quot;P1&quot; ve &quot;P2&quot; akış ünitesi en az iki pompa olduğundan emin olun ve bunları etiketlemek. Her pompa için çözücü ve reaktif giriş hatlarını belirleyin. Ikiye &quot;toplamak&quot; ve &quot;atık&quot; hatlardan çıkış hatları yerleştirin individual 100 ml şişe, sırasıyla, &quot;ürün&quot; ve &quot;atık&quot; etiketli. </li><li> Bir reaktör olarak, ısıtılmadan yeteneğine sahip bir 10 ml kapasiteli PFA bobin kullanılır. </li><li> PFA reaktör bobin girişine tüp çıkan P1 bağlayın. </li><li> Reaktör bobini sonra üç girişli polieter eter keton (PEEK) tee-karıştırıcı takın. </li><li> , Tee-miksere reaktör bobin çıkış borudan 180 ° tüp çıkan P2 bağlayın. Tee karıştırıcı üçüncü bağlantı için bir boru parça bağlayın. Bu borunun diğer ucunda, bir geri-basınç regülatörü yerleştirin. </li><li> Akış hücresinin girişine geri-basınç regülatörünün çıkışından bir hattı bağlayın. &quot;Atık / toplamak&quot; anahtarına akış hücresinin çıkışından bir hattı bağlayın. </li><li> Prime P1 ve P2 ve solventle P1 için reaktif madde hattı hem de çözücü madde hatları. Reaktif şişesine çözücü şişe P1 için belirteci hattı hareket ettirin. </li><li> P1 kullanarak, reaktör boyunca etil asetat geçmesi2 ml bobin / dakika dolana kadar o. 2 dakika boyunca / 2 ml &#39;lik bir akış oranında, P2 aracılığıyla dakika aseton geçirin. </li><li> 1 ml / dakika, her iki P1 ve P2 için çözücü akış oranlarını ayarlayın. 7 barlık bir basınç geri basınç regülatörü ayarlayın. Istenilen sıcaklığa reaktör bobin sıcaklığı ayarlayın. </li><li> Şekil 3&#39;te şematik gösterildiği gibi ekipmanları yapılandırıldığını Çift kontrol edin. </li><li> Sistem sabit sıcaklık ve basınç ulaştığında, sızıntılara karşı kontrol edin ve ardından reaksiyonu çalıştırın. </li></ol><img alt="Şekil 3," src="/files/ftp_upload/52393/52393fig3.jpg" /> Şekil reaksiyon izleme deneyleri için kullanılan ekipman yapılandırma 3. şematik. <a href="https://www.jove.com/files/ftp_upload/52393/52393fig3large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> 6. Reaksiyonu Monitör <ol><li> TakEtil asetat / aseton çözücü sisteminin adet arka tarama akış hücresi içinden geçerken. Bu, otomatik olarak sonraki tüm taramalarda düşülür. </li><li> Tarar her 15 saniyede almak için spektrometre yapılandırma (Bu durumda Raman spektrometresi, bir 10 sn entegrasyon zaman yük vagonu = 3 set ve 1 = ortalama oldu). </li><li> &quot;Reaktif&quot; etiketli bir cam şişe içinde bir kerede (0.05 mi, 0.05 mmol, 0.1 eşdeğer) piperidin enjekte edilir. </li><li> İyice gelen, anahtar P1 karıştırdıktan sonra &quot;reaktif&quot;. Için çıkış akımını &quot;çözücü&quot; olarak ayarlayın &quot;toplamak.&quot; </li><li> Tüm malzeme tamamen geri &quot;reaktifi&quot; dan, anahtar P1 &quot;çözücü&quot;. Yüklendiğinde başka bir 30 dakika süreyle reaktör bobin aracılığıyla çözücü akan devam edin. Bu süre geçtikten sonra, ısıtma kapatın. </li><li> Reaktör rulo sıcaklığı 50 ° C&#39;nin altına kadar soğutulmuş, zaman açma P1 ve P2 kapalı pompalar. </li></ol> 7. Verileri analiz <ol&gt; <li> 1608 cm-1 vs bir elektronik tablo ve arsa Raman yoğunluğu Raman spektrometresi verilerini ver zaman. </li><li> Koşulları optimize etmek için, tekrarlayıcı bir biçimde debi ve sıcaklıkları reaktör bir dizi genelinde reaksiyonu gerçekleştirin. </li><li> 1608 cm-1 vs de Raman yoğunluk Yerleşimi araziler zaman. Not: Yükseköğretim Raman yoğunluğu yüksek ürün dönüşüm ile ilişkilidir. </li></ol> 8. Optimize Kullanım Koşulları Reaksiyonu çalıştırın <ol><li> Ekranlı çeşitli koşullar (değişken akış hızları / reaktör sıcaklıkları) olması, yüksek ürün dönüşüm elde etmek için optimize koşulları kullanarak reaksiyonu çalıştırın. </li></ol> 9. Ürün izole <ol><li> Ürün balonun içindekiler alın ve buz 100 ml 2 M HCI 20 ml ihtiva eden bir beher içine dökülür. </li><li> Behere etil asetat (2 mi) ve transfer az miktarda ürün şişesi durulayın. </li><li> Buzlu karışımı karıştırıntüm buz tamamen eriyene kadar. </li><li> Bir Hirsch hunisi, yan kol şişesi, lastik yaka ve kauçuk vakum boru uzunluğunda bir filtreleme sistemi kurun. </li><li> Vakum altında elde edilen çökelti filtre, soğuk dietil eter (10 mi) ile yıkayın ve bir ısı lambası (2-3 saat) ya da kuru O / N, vakum altında kurumaya bırakın. </li><li> Çözücü olarak CDCI3 kullanılarak 1 H nükleer manyetik rezonans (NMR) spektroskopi ile ürünü tanımlamakta teyit edin. Aşağıdaki gibi 500 MHz NMR spektrometresi için, 3-acetylcoumarin 1 H-NMR verileri: δ = 2.73 (s, 3H), 7,31-7,40 (m, 2 H), 7.65 (ddd, J = 7,53, 4.37, 2.60 Hz , 2 H), 8.51 (s, 1 H) ppm, 13C NMR verileri: δ = 30.84 (CH3) 117,00 (CH) 118.56 (C), 124,86 (CH) 125,27 (CH) 130,51 (CH) 134,68 (C) 147,74 (CH) 155.64 (C), 159,52 (C) 195,77 (C) ppm. </li></ol>

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The authors have nothing to disclose.

Raman spektrometresi akış birimi ile arabirim edilebileceği hızı, reaksiyon takibinde değerli Programa doğrudan sokulan bu akla getirmektedir. Reaksiyon değişkenlerinin bir dizi kullanıcı hızlı çevrimdışı yöntemlerle olduğundan daha optimum reaksiyon koşullarında varmak için izin veren bir hızlandırılmış bir şekilde incelenebilir. Burada tarif edilen tekniklerin uygulanması, uygun bir grup bulunabilir varsayarak, yan ürünlerin oluşumunun izlenmesine olanak sağlar. Koşullar, safsızlık ...

Sürekli akış işleme kullanarak, kimyagerler onlar güvenli, etkili, ve kolaylıkla 1,2 ile kimyasal reaksiyonlar bir dizi gerçekleştirebilirsiniz bulgu vardır. Sonuç olarak, akış kimya ekipman akademik kurumlarda endüstriyel ortamlarda hem tepkileri yanı sıra araştırma laboratuarları çalıştırmak için ayrılmaz bir araç haline gelmektedir. Sentetik Kimya dönüşümler çok çeşitli akış reaktörleri 3,4 gerçekleştirilmiştir. Select durumlarda, toplu işe yaramazsa reaksiyonlar sürekli akış koşulları altında 5 sorunsuz devam ettiği gösterilmiştir. Reaksiyon optimizasyon ve kalite kontrol hem de, in-line reaksiyonu akış işleme ile izleme katılması önemli avantajlar sunmaktadır. In-line izleme gerçek örnek koşullarına gerçek zamanlı tepki ile sürekli analiz sağlar. Bu, daha hızlı ve karşılaştırılabilir off-line tekniklerine göre, bazı durumlarda, daha güvenli. In-line analiz bir dizi teknik f ile ara-yüz edilmişDüşük reaktörler 7. Örnekler enfraruj dahil 8,9, UV-görünür NMR 12,13, Raman spektroskopisi 14,15 ve kütle spektrometresi 16,17, 10,11. Araştırma grubumuz bilimsel mikrodalga birimine 18 ile Raman spektrometresi arabirim etti. Bunu kullanarak, reaksiyonların bir dizi hem nitel ve nicel 19 20 açısından izlenmektedir. Bu başarısı üzerine inşa, biz son zamanlarda bizim sürekli akış birimleri biri ile Raman spektrometresi arabirim ve anahtar ilaç gibi-ilgili organik dönüşümler bir dizi sıralı reaksiyon izlenmesi için istihdam var. 21 de izlemek mümkün olan her durumda, reaksiyonları ve aynı zamanda bir örnekte, bir kalibrasyon eğrisi ile, Raman tayf verileri elde edilen ürün dönüştürme saptayabilecektir. İşte biz aparatı kurmak ve tepkileri izlemek için nasıl kullanılacağını açıklar. Bu 3-ac piperidin-katalize sentezi ileetylcoumarin (1) burada, model reaksiyon etil asetoasetat (Şekil 1) salisilaldehit. <img alt="figür 1" src="/files/ftp_upload/52393/52393fig1.jpg" /> Şekil 1. Tabanı (1). 3-acetylcoumarin vermek üzere salisilaldehid ve etil asetoasetatın arasındaki yoğunlaşma reaksiyonu katalize <a href="https://www.jove.com/files/ftp_upload/52393/52393fig1large.jpg" target="_blank">bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a>

<table><tbody><tr><td>Salicylaldehyde</td><td>Sigma-Aldrich</td><td>S356</td><td>Reagent Grade, 98%</td></tr><tr><td>Ethyl acetoacetate</td><td>Acros Organics</td><td>117970010</td><td>99%</td></tr><tr><td>Piperidine</td><td>Sigma-Aldrich</td><td>104094</td><td>Reagent Plus, 99%</td></tr><tr><td>Hydrochloric acid</td><td>Sigma-Aldrich</td><td>320331</td><td>ACS Reagent, 37%</td></tr><tr><td>Ethyl acetate</td><td>Sigma-Aldrich</td><td>34858</td><td>CHROMASOLV, for HPLC, &gt;99.7%</td></tr><tr><td>Acetone</td><td>Sigma-Aldrich</td><td>650501</td><td>CHROMASOLV, for HPLC, &gt;99.9%</td></tr><tr><td>Flow cell</td><td>Starna Cells</td><td></td><td>583.65.65-Q-5/Z20</td></tr><tr><td>Flow unit</td><td>Vapourtec</td><td></td><td>E-series system</td></tr><tr><td>Raman spectrometer</td><td>Enwave Optronics Inc</td><td></td><td>Model EZRaman-L</td></tr></tbody></table>

real-time monitoring of reactions performed using continuous-flow processing: the preparation of 3-acetylcoumarin as an example

Sıralı izleme kullanılarak, basit ve hızlı bir şekilde sürekli akış işleme ile yerine reaksiyonları optimize etmek mümkündür. Bu teknik kullanılarak, zaman içinde tutarlı ürün kalitesinin sağlanması da mümkündür. Biz burada bir Raman spektrometresi ile piyasada bulunan bir akış birimi arayüzü nasıl gösterir. Raman akış hücresi, bu buluşta atmosfer basıncında çalıştırılabilir, yani, geri-basınç regülatörü sonra yerleştirilir. Buna ek olarak, ürün akımı akış hücresi girmeden önce bir boru uzunluğunu geçmektedir olması malzemesi, oda sıcaklığında olduğu anlamına gelir. Bu Raman sinyal yoğunluğu sıcaklığa bağlıdır çünkü spektrumları izotermal şartlarda kazanılmış olması önemlidir. Aygıt birleştirilmiş ettikten sonra, bir kimyasal reaksiyon izleme şeklini göstermektedir, salisilaldehit ve etil asetoasetat 3-acetylcoumarin piperidin-katalize sentezi bir örnek olarak kullanılır. Akış, bir oranları reaksiyonu aralığında yapılabilird sıcaklıklar, in-situ izleme aracı basit ve kolay bir şekilde koşulları optimize etmek için kullanılır.

3-acetylcoumarin sürekli akışlı hazırlanması in-line izlenmesi için temsili bir tepki olarak seçildi. Çözücü olarak etil asetat kullanarak toplu olarak, reaksiyon uygun olarak ilerlemektedir. Bununla birlikte, ürünün (1), oda sıcaklığında tam olarak çözünür değildir. Geri-basınç regülatörü potansiyel tıkanmasını önlemek, hem de sinyal edinimi karıştırmayı istiyorum akış hücresi katı parçacıklara sahip riskini azaltmak için, biz bu ve diğer reaksiyonları <sup...

Watch this Scientific Journal Video about Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example at JoVE.com

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

Gerçek zamanlı izleme sürekli akış işleme ile yerine reaksiyonların hızlı optimizasyonu sağlar. Burada 3-acetylcoumarin hazırlanması Bir örnek olarak kullanılmıştır. Reaksiyonun iyi duruma getirmek için gereken adımlar gibi in-situ Raman izleme yerine getirilmesi için aygıt tarif edilmektedir.

Sürekli akış İşleme kullanma Gerçekleştirilen Reaksiyonları Gerçek zamanlı izleme: Bir Örnek olarak 3-Acetylcoumarin Hazırlanması

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 ...

real-time-monitoring-reactions-performed-using-continuous-flow

Research

JoVE Journal

Chemistry

9.7K Görüntüleme.  University of Connecticut.  Gerçek zamanlı izleme sürekli akış işleme ile yerine reaksiyonların hızlı optimizasyonu sağlar. Burada 3-acetylcoumarin hazırlanması Bir örnek olarak kullanılmıştır. Reaksiyonun iyi duruma getirmek için gereken adımlar gibi in-situ Raman izleme yerine getirilmesi için aygıt tarif edilmektedir. 

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

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications

Methods for rapid surface immobilization of bioactive small molecules with control over orientation and immobilization density are highly desirable for biosensor and microarray applications. In this Study, we use a highly efficient covalent bioorthogonal [4+2] cycloaddition reaction between trans-cyclooctene (TCO) and 1,2,4,5-tetrazine (Tz) to enable the microfluidic immobilization of TCO/Tz-derivatized molecules. We monitor the process in real-time under continuous flow conditions using surface plasmon resonance (SPR). To enable reversible immobilization and extend the experimental range of the sensor surface, we combine a non-covalent antigen-antibody capture component with the cycloaddition reaction. By alternately presenting TCO or Tz moieties to the sensor surface, multiple capture-cycloaddition processes are now possible on one sensor surface for on-chip assembly and interaction studies of a variety of multi-component structures. We illustrate this method with two different immobilization experiments on a biosensor chip; a small molecule, AP1497 that binds FK506-binding protein 12 (FKBP12); and the same small molecule as part of an immobilized and in situ-functionalized nanoparticle.

We present a method for rapid, reversible immobilization of small molecules and functionalized nanoparticle assemblies for Surface Plasmon Resonance (SPR) studies, using sequential on-chip bioorthogonal cycloaddition chemistry and antibody-antigen capture.

Tersi Küçük Moleküller veya Biosensor Uygulamaları için Çok bileşenli Yapılar hareketsiz için microfluidic On-chip Yakalama sikloeklenme Tepki

Methods  for rapid surface immobilization of bioactive small molecules with control over  orientation and immobilization density are highly desirable for ...

Kimya

Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures

Xenobiotic metabolizing enzymes play a key function in the biotransformation of medicines and toxicants by adding functional groups that increase solubility and facilitate excretion. On some occasions those structural modifications lead to the formation of new toxic products. In order to reduce animal testing, chemical risk can be assessed using metabolically competent cells. The expression of metabolic enzymes, however, is not stable over time in many in vitro primary culture systems and is often partial or absent in cell lines. Therefore, the study of medicines, additives, and environmental pollutants metabolism in vitro should ideally be conducted in cell systems where metabolic activity has been characterized. We explain here an approach to measure the activity of a class of metabolic enzymes (Human Phase I) in 2D cell lines and primary 3D cultures using chemical probes and their metabolic products quantifiable by UPLC mass spectrometry and luminometry. The method can be implemented to test the metabolic activity in cell lines and primary cells derived from a variety of tissues.

Mass Spectrometry and Luminogenic-based Approaches to Characterize Phase I Metabolic Competency of In Vitro Cell Cultures

Metabolic competency of in vitro systems is a key requirement for the biotransformation and disposition of drugs and toxicants. In this protocol, we describe the application of reference metabolic probes to assess phase I metabolism in cell cultures.

Kütle Spektrometre ve lüminojenik tabanlı Yaklaşımlar Faz I Metabolik Yetkinlik ait Characterize<em&gt; İn Vitro</em&gt; Hücre Kültürleri

Xenobiotic metabolizing enzymes play a key function in the biotransformation of medicines and toxicants by adding functional groups that increase ...

Biyokimya

Monitoring the Reductive and Oxidative Half-Reactions of a Flavin-Dependent Monooxygenase using Stopped-Flow Spectrophotometry

Aspergillus fumigatus siderophore A (SidA) is an FAD-containing monooxygenase that catalyzes the hydroxylation of ornithine in the biosynthesis of hydroxamate siderophores that are essential for virulence (e.g. ferricrocin or N',N",N'''-triacetylfusarinine C)1. The reaction catalyzed by SidA can be divided into reductive and oxidative half-reactions (Scheme 1). In the reductive half-reaction, the oxidized FAD bound to Af SidA, is reduced by NADPH2,3. In the oxidative half-reaction, the reduced cofactor reacts with molecular oxygen to form a C4a-hydroperoxyflavin intermediate, which transfers an oxygen atom to ornithine. Here, we describe a procedure to measure the rates and detect the different spectral forms of SidA using a stopped-flow instrument installed in an anaerobic glove box. In the stopped-flow instrument, small volumes of reactants are rapidly mixed, and after the flow is stopped by the stop syringe (Figure 1), the spectral changes of the solution placed in the observation cell are recorded over time. In the first part of the experiment, we show how we can use the stopped-flow instrument in single mode, where the anaerobic reduction of the flavin in Af SidA by NADPH is directly measured. We then use double mixing settings where Af SidA is first anaerobically reduced by NADPH for a designated period of time in an aging loop, and then reacted with molecular oxygen in the observation cell (Figure 1). In order to perform this experiment, anaerobic buffers are necessary because when only the reductive half-reaction is monitored, any oxygen in the solutions will react with the reduced flavin cofactor and form a C4a-hydroperoxyflavin intermediate that will ultimately decay back into the oxidized flavin. This would not allow the user to accurately measure rates of reduction since there would be complete turnover of the enzyme. When the oxidative half-reaction is being studied the enzyme must be reduced in the absence of oxygen so that just the steps between reduction and oxidation are observed. One of the buffers used in this experiment is oxygen saturated so that we can study the oxidative half-reaction at higher concentrations of oxygen. These are often the procedures carried out when studying either the reductive or oxidative half-reactions with flavin-containing monooxygenases. The time scale of the pre-steady-state experiments performed with the stopped-flow is milliseconds to seconds, which allow the determination of intrinsic rate constants and the detection and identification of intermediates in the reaction4. The procedures described here can be applied to other flavin-dependent monooxygenases.5,6

We describe the use of a stopped-flow instrument to investigate both the reductive and oxidative half-reactions of Aspergillus fumigatus siderophore A (SidA), a flavin-dependent monooxygenase. We then show the spectra corresponding to the species in the reaction of SidA and we calculate the rate constants for their formation.

Durduruldu-Akış Spektrofotometri kullanarak Flavin-Bağımlı monooksijenaz ve indirgeyici ve Oksidatif Yarı-Tepkime İzlenmesi

Aspergillus fumigatus siderophore A (SidA) is an FAD-containing monooxygenase that catalyzes the hydroxylation of ornithine in the biosynthesis of ...

Biyomühendislik

Post Column Derivatization Using Reaction Flow High Performance Liquid Chromatography Columns

A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is presented. A major difficulty in adapting PCD to modern HPLC systems and columns is the need for large volume reaction coils that enable reagent mixing and then the derivatization reaction to take place. This large post column dead volume leads to band broadening, which results in a loss of observed separation efficiency and indeed detection in sensitivity. In reaction flow post column derivatization (RF-PCD) the derivatization reagent(s) are pumped against the flow of mobile phase into either one or two of the outer ports of the reaction flow column where it is mixed with column effluent inside a frit housed within the column end fitting. This technique allows for more efficient mixing of the column effluent and derivatization reagent(s) meaning that the volume of the reaction loops can be minimized or even eliminated altogether. It has been found that RF-PCD methods perform better than conventional PCD methods in terms of observed separation efficiency and signal to noise ratio. A further advantage of RF-PCD techniques is the ability to monitor effluent coming from the central port in its underivatized state. RF-PCD has currently been trialed on a relatively small range of post column reactions, however, there is currently no reason to suggest that RF-PCD could not be adapted to any existing one or two component (as long as both reagents are added at the same time) post column derivatization reaction.

A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is presented.

Mesaj Sütun Türevlendirme Reaksiyon Akış Yüksek Performanslı Sıvı Kromatografi sütunlar kullanma

A protocol for the use of reaction flow high performance liquid chromatography columns for methods employing post column derivatization (PCD) is ...

Temperature-programmed Deoxygenation of Acetic Acid on Molybdenum Carbide Catalysts

Temperature programmed reaction (TPRxn) is a simple yet powerful tool for screening solid catalyst performance at a variety of conditions. A TPRxn system includes a reactor, furnace, gas and vapor sources, flow control, instrumentation to quantify reaction products (e.g., gas chromatograph), and instrumentation to monitor the reaction in real time (e.g., mass spectrometer). Here, we apply the TPRxn methodology to study molybdenum carbide catalysts for the deoxygenation of acetic acid, an important reaction among many in the upgrading/stabilization of biomass pyrolysis vapors. TPRxn is used to evaluate catalyst activity and selectivity and to test hypothetical reaction pathways (e.g., decarbonylation, ketonization, and hydrogenation). The results of the TPRxn study of acetic acid deoxygenation show that molybdenum carbide is an active catalyst for this reaction at temperatures above ca. 300 &#176;C and that the reaction favors deoxygenation (i.e., C-O bond-breaking) products at temperatures below ca. 400 &#176;C and decarbonylation (i.e., C-C bond-breaking) products at temperatures above ca. 400 &#176;C.

Presented here is a protocol for the operation of a micro-scale temperature-programmed reactor for evaluating the catalytic performance of molybdenum carbide during acetic acid deoxygenation.

Molibden Karbür Katalizörler üzerinde Asetik Asit Sıcaklık programlanmış deoksijenasyonu

Temperature programmed reaction (TPRxn) is a simple yet powerful tool for screening solid catalyst performance at a variety of conditions. A TPRxn system ...

Optimization of the Ugi Reaction Using Parallel Synthesis and Automated Liquid Handling

The optimization of a Ugi reaction involving the mixing of furfurylamine, benzaldehyde, boc-glycine and t-butylisocyanide is described. Triplicate runs of 48 parallel experiments are reported, varying concentration, solvent and the excess of some of the reagents. The isolation of the product was achieved by a simple filtration and wash procedure. The highest yield obtained was 66% from 0.4 M methanol with 1.2 eq. of imine. This is significantly above the 49% yield obtained from the initial reaction under equimolar concentration at 0.4 M in methanol. Methanol solutions with reagent concentrations of 0.4M or 0.2M gave superior yields while all solvent systems at 0.07M performed poorly. At 0.2M, methanol and ethanol/methanol (60/40) mixtures were statistically equally good while THF/methanol (60/40) was poor and acetonitrile/methanol (60/40) was intermediate. Good reproducibility of the precipitate yields was obtained in these replicate experiments, allowing for subtle interaction effects to be positively identified.

The Ugi reaction has proved to be a convenient way to quickly create diverse libraries of compounds. It involves the reaction of an amine, an aldehyde, a carboxylic acid and an isonitrile typically in methanol at room temperature. In this video, we utilize a 48-slot Mettler-Toledo MiniBlock equipped with filtration tubes and a Mettler-Toledo MiniMapper automated liquid handler was used to deliver the reagents and solvent. The parameters of interest were the concentration, the solvent composition and the excess of some of the reagents.

Paralel Sentezi ve Otomatik Liquid Handling kullanma Ugi Reaksiyon optimizasyonu

The optimization of a Ugi reaction involving the mixing of furfurylamine, benzaldehyde, boc-glycine and t-butylisocyanide is described.  Triplicate runs ...

Biyoloji

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.

Sürekli akışı kimya: Diphenyldiazomethane p- Nitrobenzoic asit tepki

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

Biosynthesis of a Flavonol from a Flavanone by Establishing a One-pot Bienzymatic Cascade

Flavonols are a major subclass of flavonoids with a variety of biological and pharmacological activities. Here, we provide a method for the in vitro enzymatic synthesis of a flavonol. In this method, Atf3h and Atfls1, two key genes in the biosynthetic pathway of the flavonols, are cloned and overexpressed in Escherichia coli. The recombinant enzymes are purified via an affinity column and then a bienzymatic cascade is established in a specific synthetic buffer. Two flavonols are synthesized in this system as examples and determined by TLC and HPLC/LC/MS analyses. The method displays obvious advantages in the derivation of flavonols over other approaches. It is time- and labor-saving and highly cost-effective. The reaction is easy to be accurately controlled and thus scaled up for mass production. The target product can be purified easily due to the simple components in the system. However, this system is usually restricted to the production of a flavonol from a flavanone.

The derivation of a flavonol is crucial for its application in healthcare and the food industry. Here, we provide a detailed protocol for the biosynthesis of a flavonol from a flavanone and discuss the crucial steps and its advantages over other approaches.

Bir Flavanone bir Flavonol Biyosentezi Bir-pot Bienzymatik Cascade Kurarak

Flavonols are a major subclass of flavonoids with a variety of biological and pharmacological activities. Here, we provide a method for the in vitro ...

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

A new reaction screening technology for organic synthesis was recently demonstrated by combining elements from both continuous micro-flow and conventional batch reactors, coined stop-flow micro-tubing (SFMT) reactors. In SFMT, chemical reactions that require high pressure can be screened in parallel through a safer and convenient way. Cross-contamination, which is a common problem in reaction screening for continuous flow reactors, is avoided in SFMT. Moreover, the commercially available light-permeable micro-tubing can be incorporated into SFMT, serving as an excellent choice for light-mediated reactions due to a more effective uniform light exposure, compared to batch reactors. Overall, the SFMT reactor system is similar to continuous flow reactors and more superior than batch reactors for reactions that incorporate gas reagents and/or require light-illumination, which enables a simple but highly efficient reaction screening system. Furthermore, any successfully developed reaction in the SFMT reactor system can be conveniently translated to continuous-flow synthesis for large scale production.

A protocol for organic reaction screening using stop-flow micro-tubing (SFMT) reactors employing gaseous reactants and/or visible-light mediated reactions is presented.

Stop-akış mikro-boru reaktörler kullanımı organik dönüşümleri gelişimi için

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

A Modular Microfluidic Technology for Systematic Studies of Colloidal Semiconductor Nanocrystals

Colloidal semiconductor nanocrystals, known as quantum dots (QDs), are a rapidly growing class of materials in commercial electronics, such as light emitting diodes (LEDs) and photovoltaics (PVs). Among this material group, inorganic/organic perovskites have demonstrated significant improvement and potential towards high-efficiency, low-cost PV fabrication due to their high charge carrier mobilities and lifetimes. Despite the opportunities for perovskite QDs in large-scale PV and LED applications, the lack of fundamental and comprehensive understanding of their growth pathways has inhibited their adaptation within continuous nanomanufacturing strategies. Traditional flask-based screening approaches are generally expensive, labor-intensive, and imprecise for effectively characterizing the broad parameter space and synthesis variety relevant to colloidal QD reactions. In this work, a fully autonomous microfluidic platform is developed to systematically study the large parameter space associated with the colloidal synthesis of nanocrystals in a continuous flow format. Through the application of a novel translating three-port flow cell and modular reactor extension units, the system may rapidly collect fluorescence and absorption spectra across reactor lengths ranging 3 - 196 cm. The adjustable reactor length not only decouples the residence time from the velocity-dependent mass transfer, it also substantially improves the sampling rates and chemical consumption due to the characterization of 40 unique spectra within a single equilibrated system. Sample rates may reach up to 30,000 unique spectra per day, and the conditions cover 4 orders of magnitude in residence times ranging 100 ms - 17 min. Further applications of this system would substantially improve the rate and precision of the material discovery and screening in future studies. Detailed within this report are the system materials and assembly protocols with a general description of the automated sampling software and offline data processing.

Detailed herein are the operation and assembly protocols of a modular microfluidic screening platform for the systematic characterization of colloidal semiconductor nanocrystal syntheses. Through fully adjustable system arrangements, highly efficient spectra collection may be carried out across 4 orders of magnitude reaction time scales within a mass transfer-controlled sampling space.

Kolloidal yarı iletken Nanocrystals sistematik çalışmalar için modüler bir mikrosıvısal teknolojisi

Colloidal semiconductor nanocrystals, known as quantum dots (QDs), are a rapidly growing class of materials in commercial electronics, such as light ...