Name: preparation of expanded chitin foams and their use in the removal of aqueous copper
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The research was sponsored by the Combat Capabilities Development Command Army Research Laboratory (Cooperative Agreement Number W911NF-15-2-0020). Any opinions, findings and conclusions, or recommendations expressed in this material are those of the authors and do not necessarily reflect the views of the Army Research Lab.
We thank the Center for Advanced Materials Processing (CAMP) at Montana Technological University for the use of some of the specialized equipment required in this study. We also thank Gary Wyss, Nancy Oyer, Rick LaDouceur, John Kirtley, and Katherine Zodrow for the technical assistance and helpful discussions.

1. Preparation of expanded chitin
<ol>
	<li>Prepare a 250 mL solution of 5 wt% LiCl in dimethylacetamide (DMAc) 
	CAUTION: The solvent DMAc is a combustible irritant that may damage fertility and cause birth defects. Handle DMAc in a fume hood using chemical resistant gloves and goggles to avoid contact with skin and eyes.
	<ol>
		<li>Add 15 g of LiCl and 285 g (268 mL) of DMAc into a 500 mL Erlenmeyer flask with, then place a 50 mm Polytetrafluoroethylene (PTFE)-lined magnetic stir bar.</li>
		<li>Cap the flask w...

<ol>
	<li>Rinaudo, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chitin+and+chitosan:+Properties+and+applications.">Chitin and chitosan: Properties and applications.</a> Progress in Polymer Science. 31 (7), 603-632 (2006).</li><li>Percot, A., Viton, C., Domard, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optimization+of+chitin+extraction+from+shrimp+shells.">Optimization of chitin extraction from shrimp shells.</a> Biomacromolecules. 4 (1), 12-18 (2003).</li><li>Austin, P. 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L., Cunha, J. M., Calgaro, C. O., Tanabe, E. H., Bertuol, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surface+modification+of+chitin+using+ultrasound-assisted+and+supercritical+CO2+technologies+for+cobalt+adsorption.">Surface modification of chitin using ultrasound-assisted and supercritical CO2 technologies for cobalt adsorption.</a> Journal of Hazardous Materials. 295, 29-36 (2015).</li><li>Phongying, S., Aiba, S., Chirachanchai, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+chitosan+nanoscaffold+formation+via+chitin+whiskers.">Direct chitosan nanoscaffold formation via chitin whiskers.</a> Polymer. 48 (1), 393-400 (2007).</li><li>Tan, T. S., Chin, H. Y., Tsai, M. L., Liu, C. 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Reaction kinetics and structure modifications.</a> Carbohydrate Polymers. 12 (4), 405-418 (1990).</li><li>Scherrer, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+the+size+and+the+internal+structure+of+colloidal+particles+by+means+of+X-rays.">Determination of the size and the internal structure of colloidal particles by means of X-rays.</a> News from the Society of Sciences in G&#246;ttingen, Mathematical- Physical Class. 2, 98-100 (1918).</li><li>Brunauer, S., Emmett, P. H., Teller, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+of+gases+in+multimolecular+layers.">Adsorption of gases in multimolecular layers.</a> Journal of the American Chemical Society. 60 (2), 309-319 (1938).</li><li>Sing, K. S. 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The proposed method for chitin foam fabrication allows for the production of such foams without the need for specialized equipment or techniques. Production of the chitin foam relies on the suspension of sodium hydride within a chitin sol-gel. Contact with water from the atmosphere induces gelling of the chitin matrix and evolution of hydrogen gas by decomposition of the sodium hydride. Therefore, the critical steps of the preparation are (1) formation of the sol-gel, (2) introduction of the sodium hydride in anhydrous c...

Chitin is a mechanically robust and chemically inert biopolymer, second only to cellulose in natural abundance1. It is the major component in the exoskeleton of arthropods and in the cell walls of fungi and yeast2. Chitin is similar to cellulose, but with one hydroxyl group of each monomer replaced with an acetyl amine group (Figure 1A,B). This difference increases the strength of hydrogen bonding between adjacent polymer chains and gives chitin its characteristic structural resilience and chemical inertness2,3. Due to its properties and abundance, chitin has attracted significant industrial and academic interest. It has been studied as a scaffold for tissue growth4,5,6, as a component in composite materials7,8,9,10,11, and as a support for adsorbents and catalysts11,12,13,14. Its chemical stability, in particular, makes chitin attractive for adsorption applications that involve conditions inhospitable to common adsorbents14. In addition, the abundance of amine groups make chitin an effective adsorbent for metal ions15. However, the protonation of the amine groups under acidic conditions reduces the metal adsorption capacity of chitin16. A successful strategy is to introduce adsorption sites more resistant to protonation17,18. Instead, herein is described a simple method to increase the specific surface area and, therefore, the number of adsorption sites in chitin.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/62301/62301fig01.jpg" /> 
Figure 1. Chemical structure. (A) cellulose, (B) chitin, (C) chitosan. <a href="https://www.jove.com/files/ftp_upload/62301/62301fig01large.jpg" target="_blank">Please click here to view a larger version of this figure.</a>
In spite of its many potential uses, chitin is underutilized. Chitin processing is challenging due to its low solubility in most solvents. A key limitation to its use in catalysis and adsorption is its low specific surface area. While typical carbon and metal oxide supports have specific surface areas in the order 102-103 m2/g, commercial chitin flakes have surface areas in the order of 10 m2/g19,20,21. Methods to expand chitin into foams exist, but they invariably rely on high temperature and pressure, strong acids and bases, or specialized equipment that represent a significant entry barrier5,21,22,23,24,25. In addition, these methods tend to deacetylate chitin to form chitosan (Figure 1C)-a more soluble and reactive biopolymer5,25,26.
Herein, a method is described to expand chitin into solid foams, increase its specific surface area and adsorption capacity, and maintain its chemical integrity. The method relies on the rapid evolution of gas from within a chitin gel and requires no specialized equipment. The increased adsorption capacity of the expanded chitin is demonstrated with aqueous Cu2+-a common contaminant in the local groundwater26.
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td></td>
			<td>Unit</td>
			<td>Neat Flake</td>
			<td>Baked Foam</td>
			<td>Lyophilised Foam</td>
		</tr>
		<tr>
			<td>Crystallinity</td>
			<td>%</td>
			<td>88</td>
			<td>74</td>
			<td>58</td>
		</tr>
		<tr>
			<td>Crystal size</td>
			<td>nm</td>
			<td>6.5</td>
			<td>4.4</td>
			<td>4.4</td>
		</tr>
		<tr>
			<td>Surface Area</td>
			<td>m2/g</td>
			<td>12.6 &#177; 2.1</td>
			<td>43.1 &#177; 0.2</td>
			<td>73.9 &#177; 0.2</td>
		</tr>
		<tr>
			<td>Cu Uptake</td>
			<td>mg/g</td>
			<td>13.8 &#177; 2.9</td>
			<td>48.6 &#177; 1.9</td>
			<td>73.1 &#177; 2.0</td>
		</tr>
		<tr>
			<td>Cu Uptake</td>
			<td>atom/nm2</td>
			<td>10.5 &#177; 2.8</td>
			<td>10.7 &#177; 0.4</td>
			<td>9.4 &#177; 0.3</td>
		</tr>
	</tbody>
</table>
Table 1. Summary of material properties. Chitin foams have lower crystallinity and crystal size relative to neat chitin flakes. However, the specific surface area and Cu uptake of the chitin foams are proportionally higher than that of the neat chitin flakes.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Ammonium bicarbonate</td>
			<td>Sigma-Aldrich</td>
			<td>9830</td>
			<td>NH4HCO3, &#8805;99.5 %</td>
		</tr>
		<tr>
			<td>Chitin</td>
			<td>Sigma-Aldrich</td>
			<td>C7170</td>
			<td>Pandalus borealis, practical grade</td>
		</tr>
		<tr>
			<td>Colorimeter</td>
			<td>Hanna Instruments</td>
			<td>HI83399-01</td>
			<td>Photometer for wastewater analysis</td>
		</tr>
		<tr>
			<td>Copper High Range Checker</td>
			<td>Hanna Instruments</td>
			<td>HI702</td>
			<td>Bicinchoninate colorimetric titration</td>
		</tr>
		<tr>
			<td>Copper nitrate hydrate&#160;</td>
			<td>Sigma-Aldrich</td>
			<td>223395</td>
			<td>Cu(NO3)2 &#183; 2.5 H2O, 98 %</td>
		</tr>
		<tr>
			<td>Dimethylacetamide (DMAc)</td>
			<td>Sigma-Aldrich</td>
			<td>271012</td>
			<td>Anhydrous, 99.8 %</td>
		</tr>
		<tr>
			<td>IR Spectrophotometer</td>
			<td>Thermo Nicolet</td>
			<td>Nexus 670</td>
			<td>Fitted with an ATR cell</td>
		</tr>
		<tr>
			<td>Lithium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>310468</td>
			<td>LiCl, &#8805;99 %</td>
		</tr>
		<tr>
			<td>N2 Physisorption Apparatus</td>
			<td>Micromeritics</td>
			<td>Tristar II</td>
			<td></td>
		</tr>
		<tr>
			<td>Nitric acid</td>
			<td>BDH</td>
			<td>BDH7208-1</td>
			<td>HNO3, 0.1 N</td>
		</tr>
		<tr>
			<td>Scanning electron microscope</td>
			<td>Zeiss LEO</td>
			<td>1430 VP</td>
			<td>15 kV, secondary electron detector, 29-31 mm working distance</td>
		</tr>
		<tr>
			<td>Sodium hydride</td>
			<td>Sigma-Aldrich</td>
			<td>223441</td>
			<td>NaH, packed in mineral oil, 90 %</td>
		</tr>
		<tr>
			<td>Thermogravimetric analyzer</td>
			<td>TA Instruments</td>
			<td>Q500</td>
			<td>100 ml/min N2, 10 &#176;C/min to 800 &#176;C</td>
		</tr>
		<tr>
			<td>Water Purification System</td>
			<td>Millipore</td>
			<td>Milli-Q</td>
			<td>Type A water (18 M&#937;)</td>
		</tr>
		<tr>
			<td>X-Ray Diffractometer</td>
			<td>Rigaku</td>
			<td>Ultima IV</td>
			<td>Cu K-&#945; radiation, 8.04 keV</td>
		</tr>
	</tbody>
</table>

preparation of expanded chitin foams and their use in the removal of aqueous copper

Chitin is an underexploited, naturally abundant, mechanically robust, and chemically resistant biopolymer. These qualities are desirable in an adsorbent, but chitin lacks the necessary specific surface area, and its modification involves specialized techniques and equipment. Herein is described a novel chemical procedure for expanding chitin flakes, derived from shrimp shell waste, into foams with higher surface area. The process relies on the evolution of H2 gas from the reaction of water with NaH trapped in a chitin gel. The preparation method requires no specialized equipment. Powder X-ray diffraction and N2-physisorption indicate that the crystallite size decreases from 6.6 nm to 4.4 nm and the specific surface area increases from 12.6 &#177; 2.1 m2/g to 73.9 &#177; 0.2 m2/g. However, infrared spectroscopy and thermogravimetric analysis indicate that the process does not change the chemical identity of the chitin. The specific Cu adsorption capacity of the expanded chitin increases in proportion to specific surface area from 13.8 &#177; 2.9 mg/g to 73.1 &#177; 2.0 mg/g. However, the Cu adsorption capacity as a surface density remains relatively constant at an average of 10.1 &#177; 0.8 atom/nm2, which again suggests no change in the chemical identity of the chitin. This method offers the means to transform chitin into a higher surface area material without sacrificing its desirable properties. Although the chitin foam is described here as an adsorbent, it can be envisioned as a catalyst support, thermal insulator, and structural material.

Expanded chitin shows the same morphology regardless of the drying method. Figure 3 shows images of neat chitin flakes (Figure 3A1), oven-dried expanded chitin (Figure 3B1), and lyophilized expanded chitin (Figure&#160;3C3). While the neat flakes have the appearance of coarse sand, the expanded chitin foam has the appearance of a kernel of popped corn. Scanning electron micrographs show a similar ch...

Watch this Scientific Journal Video about Preparation of Expanded Chitin Foams and their Use in the Removal of Aqueous Copper at JoVE.com

Preparation of Expanded Chitin Foams and their Use in the Removal of Aqueous Copper

This study describes a method to expand chitin into a foam by chemical techniques that require no specialized equipment.

Chitin is an underexploited, naturally abundant, mechanically robust, and chemically resistant biopolymer. These qualities are desirable in an adsorbent, ...

This protocol allows the novel physical transformation of chitin, a high volume waste material, that is notoriously difficult to work with. The main advantage of this technique is the simplicity of the processing methodology. As it does not require specialized materials.Only the basic equipment commonly found in chemistry laboratories. We demonstrated this method with chitin, but this technique can be modified to create expanded versions of other polymers and bio polymers that form gels. In the simplest version of this technique, performed as demonstrated.Patience is important. The jelling rate will change with daily and seasonal changes to humidity. One day before preparing the expanded chitin, dry at least 1.2 grams of chitin flakes for 24 hours in an 80 degrees Celsius oven.The next morning in a fume hood and wearing chemical resistant gloves and goggles, add 15 grams of lithium chloride to 285 grams of D-MAc, In a 500 milliliter Erlenmeyer flask, containing a 50 milliliter PTFE line magnetic stir bar. Cap the flask with a rubber septum and place it on a heating stir plate. Place, a temperature probe into the mixture through the septum and stir the mixture at 400 rotations per minute at 80 degrees Celsius, for about 4 hours.When all of the lithium chloride has dissolved, add 1 gram of the oven dried chitin flakes to the 5%lithium chloride D-MAc solution. And transfer the resulting solution into a 500 milliliter round bottom flask, containing a 50 milliliter PTFE lined magnetic stir bar. Place the flask cap with the rubber septum on a stirring heat block.Pierce to septum with a needle to allow the flask to vent and heat the solution at 80 degrees Celsius, with stirring at 400 rotations per minute for 24 to 48 hours. When all of the chitin has dissolved, allow the resultant chitin sol-gel to cool to room temperature for about one hour, with stirring. Once the solution reaches room temperature, place the flask in an ice bath, with continued stirring for approximately 20 minutes, until the temperature stabilizes.To prepare a 100 milliliter slurry of sodium hydride and D-MAc. In a fume hood, wearing chemical resistant gloves and goggles, wash approximately 1 gram of sodium hydride removed from mineral oil storage. 3 times with 10 milliliters of hexane per wash.Add 0.82 grams of the wash sodium hydride to a fresh 250 milliliter Erlenmeyer flask, containing 100 milliliters of D-MAc at a PTFE line magnetic stir bar, and swirl the mixture, to produce a sodium hydride D-MAc slurry. To form the chitin gel, add the entire volume of sodium hydride slurry to the cooled sol-gel, while vigorously stirring the gel solution. Then replace the cap and continue to stir the mixture at 400 rotations per minute, until a gel forms.After the gel is formed, add 100 milliliters of deionized water to the flask in a fume hood, and remove the expanded chitin foam from the flask, breaking the foam into pieces, if necessary. Place the foam in a crystallization dish sufficiently large enough to hold the foam, along with 1000 milliliters of deionized water. Rinse the isolated gel 3 times, with 500 milliliters of deionized water per wash, before soaking the gel in 1000 milliliters of deionized water, 500 milliliters of methanol, and 1000 milliliters of fresh deionized water for 24 per immersion.After the last deionized water wash, allow the gel to air dry for 24 to 48 hours. The gel can then be dried in the oven for 48 hours at 85 degrees Celsius under ambient air or an lyophilizer, at minus 43 degrees Celsius and 0.024 millibars of pressure, for 48 hours. When a solid chitin foam is formed, use a mortar and pestle to grind the foam into a fine powder.Before drying, the knead chitin flakes, exhibit a core sand appearance. After drying, the expanded chitin morphology resembles a kernel of popcorn, regardless of the method. Scanning electron micrographs revealed that knead chitin is a compact dense structure, while the expanded chitin, resembles crinkled paper or wrinkled sheets.In x-ray diffraction studies, knead chitin displays a strong peak at 19.3 degrees corresponding to its crystal plane. Which decreases in intensity after baking, or lyophilizing. Suggesting, that drying changes the crystallinity index of the chitin.Measurement of the specific surface area obtained from nitrogen physisorption isotherms, shows the greatest uptake volume, for the expanded foams. Confirming the more open and porous structure of these samples. Despite these changes in morphology, the expansion process does not appear to affect the chemical structure of the chitin.As observed in these representative IR spectrograms. Similar observations are noted, after thermogravimetric analysis. With the onset of thermal decomposition of all three samples occurring at 260 degrees Celsius, and the maximum decomposition rate occurring at a higher temperature for chitin flakes, due to its more compact morphology.The increase in specific surface area is accompanied by an expected increase in the maximum copper uptake by chitin. However, these differences in uptake disappear, when the copper uptake is normalized by the surface area. It's important to remember that D-MAc and NAH are dangerous chemicals, that must be handled carefully.Always work in a fume hood, and wear the appropriate personal protective equipment. The limited nitrogen adsorption isotherms we collected, can only provide specific surface area. Full nitrogen absorption isotherms, can give porosity information that is critical to determining the usefulness in size-selective applications.

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Chemistry

Preparation and Use of Samarium Diiodide (SmI2) in Organic Synthesis: The Mechanistic Role of HMPA and Ni(II) Salts in the Samarium Barbier Reaction

Although initially considered an esoteric reagent, SmI2 has become a common tool for synthetic organic chemists. SmI2 is generated through the addition of molecular iodine to samarium metal in THF.1,2-3 It is a mild and selective single electron reductant and its versatility is a result of its ability to initiate a wide range of reductions including C-C bond-forming and cascade or sequential reactions. SmI2 can reduce a variety of functional groups including sulfoxides and sulfones, phosphine oxides, epoxides, alkyl and aryl halides, carbonyls, and conjugated double bonds.2-12 One of the fascinating features of SmI-2-mediated reactions is the ability to manipulate the outcome of reactions through the selective use of cosolvents or additives. In most instances, additives are essential in controlling the rate of reduction and the chemo- or stereoselectivity of reactions.13-14 Additives commonly utilized to fine tune the reactivity of SmI2 can be classified into three major groups: (1) Lewis bases (HMPA, other electron-donor ligands, chelating ethers, etc.), (2) proton sources (alcohols, water etc.), and (3) inorganic additives (Ni(acac)2, FeCl3, etc).3
Understanding the mechanism of SmI2 reactions and the role of the additives enables utilization of the full potential of the reagent in organic synthesis. The Sm-Barbier reaction is chosen to illustrate the synthetic importance and mechanistic role of two common additives: HMPA and Ni(II) in this reaction. The Sm-Barbier reaction is similar to the traditional Grignard reaction with the only difference being that the alkyl halide, carbonyl, and Sm reductant are mixed simultaneously in one pot.1,15 Examples of Sm-mediated Barbier reactions with a range of coupling partners have been reported,1,3,7,10,12 and have been utilized in key steps of the synthesis of large natural products.16,17 Previous studies on the effect of additives on SmI2 reactions have shown that HMPA enhances the reduction potential of SmI2 by coordinating to the samarium metal center, producing a more powerful,13-14,18 sterically encumbered reductant19-21 and in some cases playing an integral role in post electron-transfer steps facilitating subsequent bond-forming events.22 In the Sm-Barbier reaction, HMPA has been shown to additionally activate the alkyl halide by forming a complex in a pre-equilibrium step.23
Ni(II) salts are a catalytic additive used frequently in Sm-mediated transformations.24-27 Though critical for success, the mechanistic role of Ni(II) was not known in these reactions. Recently it has been shown that SmI2 reduces Ni(II) to Ni(0), and the reaction is then carried out through organometallic Ni(0) chemistry.28
These mechanistic studies highlight that although the same Barbier product is obtained, the use of different additives in the SmI2 reaction drastically alters the mechanistic pathway of the reaction. The protocol for running these SmI2-initiated reactions is described.

Preparation and Use of Samarium Diiodide SmI2 in Organic Synthesis: The Mechanistic Role of HMPA and NiII Salts in the Samarium Barbier Reaction

A straightforward procedure for the preparation of samarium diiodide (SmI2) in THF is described. The role of two main additives namely hexamethylphosphoramide (HMPA) and Ni(acac)2 in Sm mediated reactions is demonstrated in the Sm-Barbier reaction.

Although initially considered an esoteric reagent, SmI2 has become a common tool for synthetic organic chemists. SmI2 is generated through the addition of ...

Preparation of Hydrophobic Metal-Organic Frameworks via Plasma Enhanced Chemical Vapor Deposition of Perfluoroalkanes for the Removal of Ammonia

Plasma enhanced chemical vapor deposition (PECVD) of perfluoroalkanes has long been studied for tuning the wetting properties of surfaces. For high surface area microporous materials, such as metal-organic frameworks (MOFs), unique challenges present themselves for PECVD treatments. Herein the protocol for development of a MOF that was previously unstable to humid conditions is presented. The protocol describes the synthesis of Cu-BTC (also known as HKUST-1), the treatment of Cu-BTC with PECVD of perfluoroalkanes, the aging of materials under humid conditions, and the subsequent ammonia microbreakthrough experiments on milligram quantities of microporous materials. Cu-BTC has an extremely high surface area (~1,800 m2/g) when compared to most materials or surfaces that have been previously treated by PECVD methods. Parameters such as chamber pressure and treatment time are extremely important to ensure the perfluoroalkane plasma penetrates to and reacts with the inner MOF surfaces. Furthermore, the protocol for ammonia microbreakthrough experiments set forth here can be utilized for a variety of test gases and microporous materials.

Herein the procedures for plasma enhanced chemical vapor deposition of perfluoroalkanes on microporous materials such as metal-organic frameworks to enhance their stability and hydrophobicity are described. Furthermore, breakthrough testing of milligram quantities of samples is described in detail.

Plasma enhanced chemical vapor deposition (PECVD) of perfluoroalkanes has long been studied for tuning the wetting properties of surfaces. For high ...

Exfoliation of Egyptian Blue and Han Blue, Two Alkali Earth Copper Silicate-based Pigments

In a visualized example of the ancient past connecting with modern times, we describe the preparation and exfoliation of CaCuSi4O10 and BaCuSi4O10, the colored components of the historic Egyptian blue and Han blue pigments. The bulk forms of these materials are synthesized by both melt flux and solid-state routes, which provide some control over the crystallite size of the product. The melt flux process is time intensive, but it produces relatively large crystals at lower reaction temperatures. In comparison, the solid-state method is quicker yet requires higher reaction temperatures and yields smaller crystallites. Upon stirring in hot water, CaCuSi4O10 spontaneously exfoliates into monolayer nanosheets, which are characterized by TEM and PXRD. BaCuSi4O10 on the other hand requires ultrasonication in organic solvents to achieve exfoliation. Near infrared imaging illustrates that both the bulk and nanosheet forms of CaCuSi4O10 and BaCuSi4O10 are strong near infrared emitters. Aqueous CaCuSi4O10 and BaCuSi4O10 nanosheet dispersions are useful because they provide a new way to handle, characterize, and process these materials in colloidal form.

The preparation and exfoliation of CaCuSi4O10&#160;and BaCuSi4O10 are described. Upon stirring in hot water, CaCuSi4O10 spontaneously exfoliates into monolayers, whereas BaCuSi4O10&#160;requires ultrasonication in organic solvents. Near infrared (NIR) imaging illustrates the NIR emission properties of these materials, and aqueous dispersions of these nanomaterials are useful for solution processing.

In a visualized example of the ancient past connecting with modern times, we describe the preparation and exfoliation of CaCuSi4O10 and BaCuSi4O10, the ...

Preparation and Use of Carbonyl-decorated Carbenes in the Activation of White Phosphorus

Here we present a protocol for the synthesis of two distinct carbonyl-decorated carbenes. Both carbenes can be prepared using nearly identical procedures in multi-gram scale quantities. The goal of this manuscript is to clearly detail how to handle and prepare these unique carbenes such that a synthetic chemist of any skill level can work with them. The two carbenes described are a diamidocarbene (DAC, carbene 1) and a monoamidoaminocarbene (MAAC 2). These carbenes are highly electron-deficient and as such display reactivity profiles that are atypical of more traditional N-heterocyclic carbenes. Additionally, these two carbenes only differ in their electrophilic character and not their steric parameters, making them ideal for studying how carbene electronics influence reactivity. To demonstrate this phenomenon, we are also describing the activation of white phosphorus (P4) using these carbenes. Depending on the carbene used, two very different phosphorus-containing compounds can be isolated. When the DAC 1 is used, a tris(phosphaalkenyl)phosphane can be isolated as the exclusive product. Remarkably however, when MAAC 2 is added to P4 under identical reaction conditions, an unexpected carbene-supported P8&#160;allotrope of phosphorus is isolated exclusively. Mechanistic studies demonstrate that this carbene-supported P8allotrope forms via a [2+2] cycloaddition dimerization of a transient diphosphene which has been trapped by treatment with 2,3-dimethyl-1,3-butadiene.

Here, we present a protocol for the synthesis of two carbonyl-decorated carbenes. The protocol makes these interesting compounds readily available to chemists of all skill levels. In addition to the synthesis of these two carbenes, their use in the activation of white phosphorus is also described.

Here we present a protocol for the synthesis of two distinct carbonyl-decorated carbenes. Both carbenes can be prepared using nearly identical procedures ...

Preparation of a Corannulene-functionalized Hexahelicene by Copper(I)-catalyzed Alkyne-azide Cycloaddition of Nonplanar Polyaromatic Units

The main purpose of this video is to show 6 reaction steps of a convergent synthesis and prepare a complex molecule containing up to three nonplanar polyaromatic units, which are two corannulene moieties and a racemic hexahelicene linking them. The compound described in this work is a good host for fullerenes. Several common organic reactions, such as free-radical reactions, C-C coupling or click chemistry, are employed demonstrating the versatility of functionalization that this compound can accept. All of these reactions work for planar aromatic molecules. With subtle modifications, it is possible to achieve similar results for nonplanar polyaromatic compounds.

Preparation of a Corannulene-functionalized Hexahelicene by CopperI-catalyzed Alkyne-azide Cycloaddition of Nonplanar Polyaromatic Units

Here, we present a protocol to synthesize a complex organic compound comprised of three nonplanar polyaromatic units, assembled easily with reasonable yields.

The main purpose of this video is to show 6 reaction steps of a convergent synthesis and prepare a complex molecule containing up to three nonplanar ...

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the development of antibody drug conjugates (ADCs) and nanomedicine, fluorescent microscopy and systems chemistry. For many of these applications, specificity of the bioconjugation method used is of prime concern. The Michael addition of maleimides with cysteine(s) on the target proteins is highly selective and proceeds rapidly under mild conditions, making it one of the most popular methods for protein bioconjugation.
We demonstrate here the modification of the only surface-accessible cysteine residue on yeast cytochrome c with a ruthenium(II) bisterpyridine maleimide. The protein bioconjugation is verified by gel electrophoresis and purified by aqueous-based fast protein liquid chromatography in 27% yield of isolated protein material. Structural characterization with MALDI-TOF MS and UV-Vis is then used to verify that the bioconjugation is successful. The protocol shown here is easily applicable to other cysteine - maleimide coupling of proteins to other proteins, dyes, drugs or polymers.

Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry

This protocol details the important steps required for the bioconjugation of a cysteine containing protein to a maleimide, including reagent purification, reaction conditions, bioconjugate purification and bioconjugate characterization.

The chemical linking or bioconjugation of proteins to fluorescent dyes, drugs, polymers and other proteins has a broad range of applications, such as the ...

Visualization of Ambient Mass Spectrometry with the Use of Schlieren Photography

This manuscript outlines how to visualize mass spectrometry ambient ionization sources using schlieren photography. In order to properly optimize the mass spectrometer, it is necessary to characterize and understand the physical principles of the source. Most commercial ambient ionization sources utilize jets of nitrogen, helium, or atmospheric air to facilitate the ionization of the analyte. As a consequence, schlieren photography can be used to visualize the gas streams by exploiting the differences in refractive index between the streams and ambient air for visualization in real time. The basic setup requires a camera, mirror, flashlight, and razor blade. When properly configured, a real time image of the source is observed by watching its reflection. This allows for insight into the mechanism of action in the source, and pathways to its optimization can be elucidated. Light is shed on an otherwise invisible situation.

This paper presents a protocol for the visualization of gaseous streams of an ambient ionization source using schlieren photography and mass spectrometry.

This manuscript outlines how to visualize mass spectrometry ambient ionization sources using schlieren photography. In order to properly optimize the mass ...

Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single molecule level. An electrostatic self-assembly and covalent fixation (ESA-CF) process is used for the synthesis of the cyclic poly(tetrahydrofuran) (poly(THF)). The diffusive motion of individual cyclic polymer chains in a melt state is visualized using single molecule fluorescence imaging by incorporating a fluorophore unit in the cyclic chains. The diffusive motion of the chains is quantitatively characterized by means of a combination of mean-squared displacement (MSD) analysis and a cumulative distribution function (CDF) analysis. The cyclic polymer exhibits multiple-mode diffusion which is distinct from its linear counterpart. The results demonstrate that the diffusional heterogeneity of polymers that is often hidden behind ensemble averaging can be revealed by the efficient synthesis of the cyclic polymers using the ESA-CF process and the quantitative analysis of the diffusive motion at the single molecule level using the MSD and CDF analyses.

A protocol for the synthesis and characterization of diffusive motion of cyclic polymers at the single molecule level is presented.

We demonstrate a method for the synthesis of cyclic polymers and a protocol for characterizing their diffusive motion in a melt state at the single ...

Preparation of Chitosan-based Injectable Hydrogels and Its Application in 3D Cell Culture

The protocol presents a facile, efficient, and versatile method to prepare chitosan-based hydrogels using dynamic imine chemistry. The hydrogel is prepared by mixing solutions of glycol chitosan with a synthesized benzaldehyde terminated polymer gelator, and hydrogels are efficiently obtained in several minutes at room temperature. By varying ratios between glycol chitosan, polymer gelator, and water contents, versatile hydrogels with different gelation times and stiffness are obtained. When damaged, the hydrogel can recover its appearances and modulus, due to the reversibility of the dynamic imine bonds as crosslinkages. This self-healable property enables the hydrogel to be injectable since it can be self-healed from squeezed pieces to an integral bulk hydrogel after the injection process. The hydrogel is also multi-responsive to many bio-active stimuli due to different equilibration statuses of the dynamic imine bonds. This hydrogel was confirmed as bio-compatible, and L929 mouse fibroblast cells were embedded following standard procedures and the cell proliferation was easily assessed by a 3D cell cultivation process. The hydrogel can offer an adjustable platform for different research where a physiological mimic of a 3D environment for cells is profited. Along with its multi-responsive, self-healable, and injectable properties, the hydrogels can potentially be applied as multiple carriers for drugs and cells in future bio-medical applications.

Here we describe a facile preparation of chitosan-based injectable hydrogels using dynamic imine chemistry. Methods to adjust the hydrogel&#8217;s mechanical strength and its application in 3D cell culture are presented.

The protocol presents a facile, efficient, and versatile method to prepare chitosan-based hydrogels using dynamic imine chemistry. The hydrogel is ...

A Convenient Method for Extraction and Analysis with High-Pressure Liquid Chromatography of Catecholamine Neurotransmitters and Their Metabolites

The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and related diseases, but their precise measurement is still a challenge. Many protocols have been described for neurotransmitter measurement by a variety of instruments, including high-pressure liquid chromatography (HPLC). However, there are shortcomings, such as complicated operation or hard-to-detect multiple targets, which cannot be avoided, and presently, the dominant analysis technique is still HPLC coupled with sensitive electrochemical or fluorimetric detection, due to its high sensitivity and good selectivity. Here, a detailed protocol is described for the pretreatment and detection of catecholamines with high pressure liquid chromatography with electrochemical detection (HPLC-ECD) in real urine samples of infants, using electrospun composite nanofibers composed of polymeric crown ether with polystyrene as adsorbent, also known as the packed-fiber solid phase extraction (PFSPE) method. We show how urine samples can be easily precleaned by a nanofiber-packed solid phase column, and how the analytes in the sample can be rapidly enriched, desorbed, and detected on an ECD system. PFSPE greatly simplifies the pretreatment procedures for biological samples, allowing for decreased time, expense, and reduction of the loss of targets. 
Overall, this work illustrates a simple and convenient protocol for solid-phase extraction coupled to an HPLC-ECD system for simultaneous determination of three monoamine neurotransmitters (norepinephrine (NE), epinephrine (E), dopamine (DA)) and two of their metabolites (3-methoxy-4-hydroxyphenylglycol (MHPG) and 3,4-dihydroxy-phenylacetic acid (DOPAC)) in infants' urine. The established protocol was applied to assess the differences of urinary catecholamines and their metabolites between high-risk infants with perinatal brain damage and healthy controls. Comparative analysis revealed a significant difference in urinary MHPG between the two groups, indicating that the catecholamine metabolites may be an important candidate marker for early diagnosis of cases at risk for brain damage in infants.

We present a convenient solid-phase extraction coupled to high-pressure liquid chromatography (HPLC) with electrochemical detection (ECD) for simultaneous determination of three monoamine neurotransmitters and two of their metabolites in infants' urine. We also identify the metabolite MHPG as a potential biomarker for the early diagnosis of brain damage for infants.

The extraction and analysis of catecholamine neurotransmitters in biological fluids is of great importance in assessing nervous system function and ...