Name: sulfate separation by selective crystallization with a bis-iminoguanidinium ligand
Uploaded: 2016-09-08T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Sulfate Separation by Selective Crystallization with a Bis-iminoguanidinium Ligand at JoVE.com

This work was supported by the U.S. Department of Energy, Office of Science, Basic Energy Sciences, Chemical Sciences, Geosciences, and Biosciences Division. We thank the University of North Carolina Wilmington for providing the seawater.

1. Synthesis of 1,4-Benzene-bis(iminoguanidinium) Chloride (BBIG-Cl)
<ol>
	<li>In Situ Synthesis of the 1,4-Benzene-bis(iminoguanidinium) Chloride Ligand (BBIG-Cl) and Its Crystallization with Sulfate
	<ol>
		<li>Add 0.067 g of terephthalaldehyde and 2.2 ml of a 0.5 M aqueous solution of aminoguanidinium chloride to 10 ml of deionized water in a 25 ml round bottom flask equipped with a magnetic stir bar.</li>
		<li>Stir the solution magnetically for four hours at 20 &#176;C. This will yield a slightly yellow solution of BBIG-Cl.</li>
		<li>Add 0.5 ml of a 1 M aqueous solution of sodium sulfate. This will result in the instant precipitation of BBIG-SO4 as a crystalline white solid.</li>
		<li>Filter the solid using vacuum filtration to recover BBIG-SO4. Wash the solid on the filter paper five times with 5 ml aliquots of water in order to obtain the pure sulfate salt.</li>
		<li>Check the phase purity of the crystalline BBIG-SO4 obtained by powder X-ray diffraction12. Compare with the pattern shown in Figure 1.</li>
	</ol>
	</li>
	<li>Ex Situ Synthesis of 1,4-Benzene-bis(iminoguanidinium) Chloride11
	<ol>
		<li>Add 4 g of terephthalaldehyde and 7.26 g of aminoguanidinium chloride to 20 ml of ethanol in a 50 ml round bottom flask equipped with a magnetic stir bar.</li>
		<li>Heat the solution to 60 &#176;C using a hotplate, and stir with a magnetic stir bar for 2 hr. Cool the solution to 20 &#176;C and let it sit for 3 hr, then collect the solid by vacuum filtration through a filter-paper equipped B&#252;chner funnel.</li>
		<li>Suspend the obtained solid in 20 ml of ethanol and heat on a hotplate until boiling. If the solid does not go completely into solution at this point, add small aliquots (1 ml) of ethanol, allowing each time the solution to reach boiling temperature, until all solid is dissolved.</li>
		<li>Allow the flask to cool to room temperature, then place in a 0 &#176;C freezer overnight. Collect the solid by filtering through a filter-paper equipped B&#252;chner funnel using vacuum filtration.</li>
		<li>Confirm the identity and purity of BBIG-Cl by 1H NMR spectroscopy13. Compare with the spectrum shown in Figure 2.</li>
	</ol>
	</li>
</ol>
2. Sulfate Separation from Seawater
<ol>
	<li>Sulfate crystallization as BBIG-SO4 
	NOTE: The amount of BBIG-Cl necessary to remove the sulfate depends on the exact amount of sulfate in the seawater. It was found that using 1.5 equivalents of BBIG-Cl relative to sulfate results in 99% removal of sulfate. The seawater used in this protocol has a concentration of 30 mM sulfate, as determined by titration with BaCl2.
	<ol>
		<li>Filter the seawater with a 0.22 &#956;m syringe filter or filtration membrane with small pore size to remove suspended particulates and bio organisms.</li>
		<li>Make a 30 mM solution of BBIG-Cl using deionized water and solid BBIG-Cl prepared as described in the previous section.</li>
		<li>Add the BBIG-Cl solution to the seawater in a 1.5:1 (v/v) proportion.</li>
		<li>Stir the mixture for a few hours to ensure quantitative (&#62; 99%) removal of sulfate.</li>
		<li>Collect the solid by filtering through a filter-paper equipped B&#252;chner funnel using vacuum filtration. Wash the solid on the filter paper five times with 5 ml aliquots of water.</li>
		<li>Dry the isolated solid under vacuum and weigh it to determine the yield.</li>
	</ol>
	</li>
	<li>Ligand Recovery
	<ol>
		<li>Add 53.1 mg of BBIG-SO4 to a 2 ml solution of NaOH (10%) in a 20 ml scintillation vial equipped with a magnetic stir bar.</li>
		<li>Stir the mixture for two hours at 20 &#176;C. A slightly yellow precipitate will form.</li>
		<li>Filter the solid through a filter-paper equipped B&#252;chner funnel using vacuum filtration. Wash the solid on the filter paper with 0.2 ml of water, and dry under vacuum.</li>
		<li>Characterize the recovered solid by NMR13 to confirm its identity as the bis(guanidine) free base. Compare with the NMR spectrum shown in Figure 3.</li>
	</ol>
	</li>
	<li>Determination of the Amount of Sulfate Removed from Seawater by &#946; Liquid Scintillation Counting 
	CAUTION: This technique involves the use of radioisotopes, which pose a different class of hazards than what is normally encountered in most labs. Special radiation protection equipment is usually required when handling the radionuclides. Thus, it is essential that the procedure is followed carefully and that a safety officer is consulted for advice and guidance.
	<ol>
		<li>Calculate the volume of the stock solution of the sulfur-35 radioisotope (5 mCi/ml) used to ensure there is more than 5 million counts per minute (cpm) per milliliter of seawater solution, using the following equations (cpm and curies (Ci) are both units of measure for radioactivity): 
		<img alt="Equation 1" src="/files/ftp_upload/54411/54411eq1.jpg" /> 
		<img alt="Equation 2" src="/files/ftp_upload/54411/54411eq2.jpg" /> 
		<img alt="Equation 3" src="/files/ftp_upload/54411/54411eq3.jpg" /></li>
		<li>Spike 25 ml of the seawater with 0.0112 ml of 5.0 mCi/ml solution of 35S radiolabeled sodium sulfate solution.</li>
		<li>Prepare 0, 15, 30, 33, 45, and 60 mM solutions of BBIG-Cl in deionized water and combine 0.750 ml of these solutions with an equal volume of 35S-radiolabeled sulfate spiked seawater in a 2 ml centrifuge tube.</li>
		<li>Stir the mixture via a rotating wheel or vortex in an incubator/air-box maintained at a constant temperature of 25 &#177; 0.2 &#176;C for 24 hr.</li>
		<li>Centrifuge the solutions at 1,500 x g for 10 min at 25 &#176;C.</li>
		<li>After centrifugation, remove 1.2 ml of each solution using a syringe, then filter it through a 0.22 &#956;m syringe filter to remove the suspended precipitate. Pipette 1.0 ml from each of these solutions into 20 ml of scintillation cocktail in polypropylene scintillation vials. The solution containing no BBIG-Cl (the control solution) should be diluted ten-fold with deionized water prior to addition to the scintillation cocktail.</li>
		<li>Place the scintillation vials containing the samples and the scintillation cocktail on a liquid scintillation counter and let it sit for 1 hr prior to counting to allow the samples to dark-adapt. 
		NOTE: Prior to counting the samples, calibrate the instrument and allow each sample to count for 30 min. Count additional vials containing only scintillation cocktail in order to allow for a background correction that is used when determining the concentrations of sulfate in solution.</li>
		<li>Determine the amount of sulfate removed, using the following equations: 
		<img alt="Equation 4" src="/files/ftp_upload/54411/54411eq4.jpg" /> 
		<img alt="Equation 5" src="/files/ftp_upload/54411/54411eq5.jpg" /> 
		<img alt="Equation 6" src="/files/ftp_upload/54411/54411eq6.jpg" /></li>
	</ol>
	</li>
</ol>

<ol>
	<li>Langton, M. L., Serpell, C. J., Beer, P. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anion+Recognition+in+Water:+Recent+Advances+from+Supramolecular+and+Macromolecular+Perspective.">Anion Recognition in Water: Recent Advances from Supramolecular and Macromolecular Perspective.</a> Angew. Chem. Int. Ed. 55, 1974-1987 (2016).</li><li>Busschaert, N., Caltagirone, C., Van Rossom, W., Gale, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+Supramolecular+Anion+Recognition.">Applications of Supramolecular Anion Recognition.</a> Chem. Rev. 115, 8038-8155 (2015).</li><li>Moyer, B. A., Custelcean, R., Hay, B. P., Sessler, J. L., Bowman-James, K., Day, V. W., Sung-Ok, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Case+for+Molecular+Recognition+in+Nuclear+Separations:+Sulfate+Separation+from+Nuclear+Wastes.">A Case for Molecular Recognition in Nuclear Separations: Sulfate Separation from Nuclear Wastes.</a> Inorg. Chem. 52, 3473-3490 (2013).</li><li>Kim, S. K., Lee, J., Williams, N. J., Lynch, V. M., Hay, B. P., Moyer, B. A., Sessler, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bipyrrole-Strapped+Calix[4]pyrroles:+Strong+Anion+Receptors+That+Extract+the+Sulfate+Anion.">Bipyrrole-Strapped Calix[4]pyrroles: Strong Anion Receptors That Extract the Sulfate Anion.</a> J. Am. Chem. Soc. 136, 15079-15085 (2014).</li><li>Jia, C., Wu, B., Li, S., Huang, X., Zhao, Q., Li, Q., Yang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+Efficient+Extraction+of+Sulfate+Ions+with+a+Tripodal+Hexaurea+Receptor.">Highly Efficient Extraction of Sulfate Ions with a Tripodal Hexaurea Receptor.</a> Angew. Chem. Int. Ed. 50, 486-490 (2011).</li><li>Rajbanshi, A., Moyer, B. A., Custelcean, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sulfate+Separation+from+Aqueous+Alkaline+Solutions+by+Selective+Crystallization+of+Alkali+Metal+Coordination+Capsules.">Sulfate Separation from Aqueous Alkaline Solutions by Selective Crystallization of Alkali Metal Coordination Capsules.</a> Cryst. Growth Des. 11, 2702-2706 (2011).</li><li>Custelcean, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urea-Functionalized+Crystalline+Capsules+for+Recognition+and+Separation+of+Tetrahedral+Oxoanions.">Urea-Functionalized Crystalline Capsules for Recognition and Separation of Tetrahedral Oxoanions.</a> Chem. Commun. 49, 2173-2182 (2013).</li><li>Custelcean, R., Sloop, F. V., Rajbanshi, A., Wan, S., Moyer, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sodium+Sulfate+Separation+from+Aqueous+Alkaline+Solutions+via+Crystalline+Urea-Functionalized+Capsules:+Thermodynamics+and+Kinetics+of+Crystallization.">Sodium Sulfate Separation from Aqueous Alkaline Solutions via Crystalline Urea-Functionalized Capsules: Thermodynamics and Kinetics of Crystallization.</a> Cryst. Growth Des. 15, 517-522 (2015).</li><li>Custelcean, R., Williams, N. J., Seipp, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aqueous+Sulfate+Separation+by+Crystallization+of+Sulfate-Water+Clusters.">Aqueous Sulfate Separation by Crystallization of Sulfate-Water Clusters.</a> Angew. Chem. Int. Ed. 54, 10525-10529 (2015).</li><li>Custelcean, R., Williams, N. J., Seipp, C. A., Ivanov, A. S., Bryantsev, V. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aqueous+Sulfate+Separation+by+Sequestration+of+[(SO4)(H2O)4]4-+Clusters+within+Highly+Insoluble+Imine-Linked+Bis-Guanidinium+Crystals.">Aqueous Sulfate Separation by Sequestration of [(SO4)(H2O)4]4- Clusters within Highly Insoluble Imine-Linked Bis-Guanidinium Crystals.</a> Chem. Eur. J. 22, 1997-2003 (2016).</li><li>Khownium, K., Wood, S. J., Miller, K. A., Balakrishna, R., Nguyen, T. B., Kimbrell, M. R., Georg, G. I., David, S. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+Endotoxin-Sequestering+Compounds+with+Terephthaldehyde-bis-guanylhydrazone+Scaffolds.">Novel Endotoxin-Sequestering Compounds with Terephthaldehyde-bis-guanylhydrazone Scaffolds.</a> Bioorg. Med. Chem. Lett. 16, 1305-1308 (2006).</li><li>Pecharsky, V. K., Zavalij, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fundamentals of Powder Diffraction and Structural Characterization of Materials. , (2005).</li><li>Goldenberg, D. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Principles of NMR Spectroscopy: An Illustrated Guide. , (2016).</li></ol>

The authors have nothing to disclose.

This technique is rather tolerant to many deviations from the written procedure, which makes it quite robust. There are however two critical steps that must be followed. First, the BBIG-Cl ligand needs to be as pure as possible. Impurities will not only affect the crystallization and the solubility of the resulting sulfate salt, but will also make it difficult to calculate the amount required for quantitative sulfate removal from solution. Second, all steps in the &#946; liquid scintillation counting section need to be followed meticulously, as this technique may be very sensitive to subtle changes.
Due to the simplicity of the crystallization technique, troubleshooting will most likely not be needed. Some common issues are discussed as follows. In the case that the BBIG-Cl ligand does not appear to be removing the sulfate present in solution, one of two issues is the most likely culprit. If using the ex situ synthesized BBIG-Cl, confirm its identity and purity. Take an 1H-NMR of the starting material and compare it with the reference spectrum in Figure 2. Another common culprit of this problem is the pH of the solution. If synthesizing the BBIG-Cl ligand in situ, make sure the pH of the solution is slightly acidic (pH = 5-6). Due to the fact that the active species is the protonated ligand, the method is sensitive to the solution pH. Basic solutions will deprotonate the guanidinium groups, yielding a neutral ligand that is incapable of crystallizing sulfate. If the pH is basic, a simple adjustment with HCl to a pH of about 5-6 will provide the optimal conditions for quantitative sulfate removal. This problem does highlight one of the main limitations of this technique, in that it is unable to remove sulfate from basic solutions. However, the BBIG ligand is quite acid-stable, so adjusting the pH of the solution provides a simple remedy to this problem. Another variable that may affect the sulfate crystallization efficiency is the ionic strength of the solution. While sulfate separation from seawater proved very efficient, it is possible the yield of sulfate separation to be lower when this method is applied to solutions with very high ionic strength.
The technique demonstrated in this protocol is extremely efficient, selective, green, and cost effective. By comparison, alternative sulfate removal methods involve expensive and high-maintenance membranes or ion exchange columns with low separation selectivity.3 Furthermore, compared to existing methods, the technique presented here is very simple and requires little technical knowledge and experience in separation chemistry.
This crystallization technique offers a general approach to quantitative removal of sulfate from aqueous solutions. While seawater was used in this protocol to demonstrate the technique, this crystallization method is not limited to seawater, and could be used for sulfate removal from virtually any aqueous solutions. Since the class of bis(iminoguanidinium) ligands employed can be easily synthesized in one step from readily available dialdehyde and aminoguanidinium precursors, there are potentially many other simple combinations that may result in an effective precipitation agent for sulfate or other oxoanions. Thus, mastering the techniques presented in this protocol will enable one to develop his/her own crystallization ligand with potentially even better selectivity and efficacy than the BBIG ligand presented here.

Selective separation of hydrophilic oxoanions (e.g., sulfate, chromate, phosphate) from competitive aqueous solutions represents a fundamental challenge with relevance to environmental remediation, energy production, and human health.1,2 Sulfate in particular is difficult to extract from water due to its intrinsic reluctance to shed its hydration sphere and migrate into less polar environments.3 Making aqueous sulfate extraction more efficient typically requires complex receptors that are difficult and tedious to synthesize and purify, often involving toxic reagents and solvents.4,5
Selective crystallization offers a simple yet effective alternative to sulfate separation from water.6-9 Though some metal cations such as Ba2+, Pb2+, or Ra2+ form very insoluble sulfate salts, their use in sulfate separation is not always practical due to their high toxicity and sometimes-low selectivity. Employing organic ligands as sulfate precipitants takes advantage of the structural diversity and amenability to design characteristic to organic molecules. An ideal organic ligand for aqueous sulfate crystallization should be soluble in water, yet form an insoluble sulfate salt or complex in a relatively short time and in the presence of high concentrations of competing ions. Additionally, it should be easy to synthesize and recycle. One such a ligand, 1,4-benzene-bis(iminoguanidinium) (BBIG), self-assembled in situ from two commercially available precursors, terephthalaldehyde and aminoguanidinium chloride, was recently found to be extremely effective in aqueous sulfate separation.10 The ligand is water-soluble in the chloride form, and selectively crystallizes with sulfate into an extremely insoluble salt that can be easily removed from solution by simple filtration. The BBIG ligand can then be recovered by deprotonation with aqueous NaOH and crystallization of the neutral bis-iminoguanidine, which can be converted back into the chloride form with aqueous HCl, and reused in another separation cycle. The efficacy of this ligand in removing sulfate from water is so great that monitoring the remaining sulfate concentration in solution is no longer a trivial task, requiring a more advanced technique that allows accurate measurement of trace amounts of the anion. For this purpose, radiolabeled 35S sulfate tracer in conjunction with &#946; liquid scintillation counting was employed, a technique commonly utilized in liquid-liquid extractive separations, and recently demonstrated to be effective in monitoring sulfate crystallization.8
This protocol demonstrates the one-pot in situ synthesis of the BBIG ligand and its crystallization as the sulfate salt from aqueous solutions. The ex situ synthesis of the ligand11 is also presented as a convenient method for the production of larger amounts of BBIG-Cl, which can be stored in the crystalline form until ready to use. Sulfate removal from seawater using the previously prepared BBIG-Cl ligand is then demonstrated. Finally, the use of 35S-labeled sulfate and &#946; liquid scintillation counting for measuring the sulfate concentration in seawater is demonstrated. This protocol is intended to provide a tutorial for those broadly interested in exploring the use of selective crystallization for aqueous anion separation.

<table><tbody><tr><td>Terephthalaldehyde</td><td>Sigma</td><td>T2207</td><td></td></tr><tr><td>Aminoguanidinium Chloride</td><td>Sigma</td><td>#396494</td><td></td></tr><tr><td>Sodium Sulfate</td><td>Sigma</td><td>#239313</td><td></td></tr><tr><td>Barium Chloride</td><td>Sigma</td><td>#342920</td><td>Highly Toxic</td></tr><tr><td>Ethanol</td><td>Any</td><td></td><td>Reagent Grade (190 proof)</td></tr><tr><td>Sodium Hydroxide</td><td>EMD</td><td>SX0590-1</td><td></td></tr><tr><td>Hydrochloric Acid</td><td>Sigma</td><td>#258148</td><td></td></tr><tr><td>Filter Paper</td><td>Any</td><td>-</td><td>Any qualitative or analytical filter paper will work</td></tr><tr><td>Syringe Filter (0.22 &amp;#956;m)</td><td>Any</td><td>-</td><td>Nylon filter</td></tr><tr><td>&lt;sup&gt;35&lt;/sup&gt;S Labeled Sulfate</td><td>Perkin Elmer</td><td>NEX041005MC</td><td></td></tr><tr><td>Ultima Gold Scintillation Cocktail</td><td>Perkin Elmer</td><td>#6013329</td><td></td></tr><tr><td>Polypropylene Vials&amp;nbsp;</td><td>Any</td><td>-</td><td></td></tr><tr><td>Disposable Syringe (2-3 ml)</td><td>Any</td><td>-</td><td>Any disposable plastic syringe works</td></tr></tbody></table>

sulfate separation by selective crystallization with a bis-iminoguanidinium ligand

A simple and effective method for selective sulfate separation from aqueous solutions by crystallization with a bis-guanidinium ligand, 1,4-benzene-bis(iminoguanidinium) (BBIG), is demonstrated. The ligand is synthesized as the chloride salt (BBIG-Cl) by in situ imine condensation of terephthalaldehyde with aminoguanidinium chloride in water, followed by crystallization as the sulfate salt (BBIG-SO4). Alternatively, BBIG-Cl is synthesized ex situ in larger scale from ethanol. The sulfate separation ability of the BBIG ligand is demonstrated by selective and quantitative crystallization of sulfate from seawater. The ligand can be recycled by neutralization of BBIG-SO4 with aqueous NaOH and crystallization of the neutral bis-iminoguanidine, which can be converted back into BBIG-Cl with aqueous HCl and reused in another separation cycle. Finally, 35S-labeled sulfate and &#946; liquid scintillation counting are employed for monitoring the sulfate concentration in solution. Overall, this protocol will instruct the user in the necessary skills to synthesize a ligand, employ it in the selective crystallization of sulfate from aqueous solutions, and quantify the separation efficiency.

The powder X-ray diffraction pattern of BBIG-SO4 (Figure 1) allows for unambiguous confirmation of the identity of the crystallized solid. In comparing the obtained pattern versus the reference one, peak intensity matters less than peak positioning. All strong peaks shown in the reference should be present in the obtained sample. The appearance of strong peaks in the sample that are absent in the reference pattern indicates the presence of impurities.
1H-NMR of BBIG-Cl and the recovered ligand (Figures 2 and 3) enable an assessment of both the identity of the compounds as well as their purity to about 5%. Comparison with these spectra help to ensure that the ligand was fully formed and that any impurities were adequately removed during the filtrations and/or recrystallizations. In comparing the obtained spectrum versus the reference, it is important to make sure all peaks are present in the exact positions shown. Use the same solvents used in the reference spectra so that the relative shift of the peaks do not change.
The results of sulfate separation from seawater are shown in Table 1, with over 99% of the sulfate being removed using only 1.5 molar equivalents of BBIG-Cl. This represents near-quantitative removal of sulfate from seawater despite the high ionic strength of the medium, demonstrating the efficacy of the technique described.
BBIG-Cl was obtained in a 70% yield via the ex situ method, while BBIG-SO4 was obtained in 86% yield via the in situ synthesis of BBIG-Cl. The ligand recovery yield was 93%. All organic reactions carried out in this procedure are high yielding and operationally simple, making the compounds easily accessible even to a novice chemist.
<img alt="Figure 1" src="/files/ftp_upload/54411/54411fig1.jpg" /> 
Figure 1: Powder X-ray diffraction pattern of BBIG-SO4. The pattern was obtained with a powder X-ray diffractometer using a flat sample stage in reflection mode. The strongest peaks are marked in red. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54411/54411fig1large.jpg">Please click here to view a larger version of this figure.</a>
<img alt="Figure 2" src="/files/ftp_upload/54411/54411fig2.jpg" /> 
Figure 2: 1H-NMR spectrum of BBIG-Cl. The spectrum was taken in DMSO-d6 with a 400 MHz NMR instrument. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54411/54411fig2large.jpg">Please click here to view a larger version of this figure.</a>
<img alt="Figure 3" src="/files/ftp_upload/54411/54411fig3.jpg" /> 
Figure 3: 1H-NMR spectrum of the recovered BBIG ligand. The spectrum was taken in MeOD with a 400 MHz NMR instrument. <a href="https://www-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54411/54411fig3large.jpg">Please click here to view a larger version of this figure.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>BBIG equiv</td>
			<td>[Sulfate] left (mM)</td>
			<td>Sulfate removed (%)</td>
		</tr>
		<tr>
			<td>1</td>
			<td>3.5</td>
			<td>88</td>
		</tr>
		<tr>
			<td>1.1</td>
			<td>1.6</td>
			<td>95</td>
		</tr>
		<tr>
			<td>1.5</td>
			<td>0.3</td>
			<td>99</td>
		</tr>
		<tr>
			<td>2</td>
			<td>0.3</td>
			<td>99</td>
		</tr>
	</tbody>
</table>
Table 1: Representative results from sulfate separation from seawater. The data shows removal of up to 99% of sulfate from seawater using only 1.5 M equivalents of BBIG-Cl. The initial sulfate concentration in seawater was 30 mM.

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Sulfate Separation by Selective Crystallization with a Bis-iminoguanidinium Ligand

A protocol for in situ aqueous synthesis of a bis(iminoguanidinium) ligand and its utilization in selective separation of sulfate is presented.

A simple and effective method for selective sulfate separation from aqueous solutions by crystallization with a bis-guanidinium ligand, ...

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8.3K Views.  Oak Ridge National Laboratory.  Een protocol voor in situ synthese van een waterige bis (iminoguanidinium) ligand en het gebruik ervan in selectieve scheiding van sulfaat wordt gepresenteerd. 

Video: Sulfate Separation by Selective Crystallization with a Bis-iminoguanidinium Ligand

Synthesis and Purification of Iodoaziridines Involving Quantitative Selection of the Optimal Stationary Phase for Chromatography

The highly diastereoselective preparation of cis-N-Ts-iodoaziridines through reaction of diiodomethyllithium with N-Ts aldimines is described. Diiodomethyllithium is prepared by the deprotonation of diiodomethane with LiHMDS, in a THF/diethyl ether mixture, at -78 &#176;C in the dark. These conditions are essential for the stability of the LiCHI2 reagent generated. The subsequent dropwise addition of N-Ts aldimines to the preformed diiodomethyllithium solution affords an amino-diiodide intermediate, which is not isolated. Rapid warming of the reaction mixture to 0 &#176;C promotes cyclization to afford iodoaziridines with exclusive cis-diastereoselectivity. The addition and cyclization stages of the reaction are mediated in one reaction flask by careful temperature control.
Due to the sensitivity of the iodoaziridines to purification, assessment of suitable methods of purification is required. A protocol to assess the stability of sensitive compounds to stationary phases for column chromatography is described. This method is suitable to apply to new iodoaziridines, or other potentially sensitive novel compounds. Consequently this method may find application in range of synthetic projects. The procedure involves firstly the assessment of the reaction yield, prior to purification, by 1H NMR spectroscopy with comparison to an internal standard. Portions of impure product mixture are then exposed to slurries of various stationary phases appropriate for chromatography, in a solvent system suitable as the eluent in flash chromatography. After stirring for 30 min to mimic chromatography, followed by filtering, the samples are analyzed by 1H NMR spectroscopy. Calculated yields for each stationary phase are then compared to that initially obtained from the crude reaction mixture. The results obtained provide a quantitative assessment of the stability of the compound to the different stationary phases; hence the optimal can be selected. The choice of basic alumina, modified to activity IV, as a suitable stationary phase has allowed isolation of certain iodoaziridines in excellent yield and purity.

A protocol for the diastereoselective one-pot preparation of cis-N-Ts-iodoaziridines is described. The generation of diiodomethyllithium, addition to N-Ts aldimines and cyclization of the amino gem-diiodide intermediate to iodoaziridines is demonstrated. Also included is a protocol to rapidly and quantitatively assess the most appropriate stationary phase for purification by chromatography.

The highly diastereoselective preparation of cis-N-Ts-iodoaziridines through reaction of diiodomethyllithium with N-Ts aldimines is described. ...

A Direct, Early Stage Guanidinylation Protocol for the Synthesis of Complex Aminoguanidine-containing Natural Products

The guanidine functional group, displayed most prominently in the amino acid arginine, one of the fundamental building blocks of life, is an important structural element found in many complex natural products and pharmaceuticals. Owing to the continual discovery of new guanidine-containing natural products and designed small molecules, rapid and efficient guanidinylation methods are of keen interest to synthetic and medicinal organic chemists. Because the nucleophilicity and basicity of guanidines can affect subsequent chemical transformations, traditional, indirect guanidinylation is typically pursued. Indirect methods commonly employ multiple protection steps involving a latent amine precursor, such as an azide, phthalimide, or carbamate. By circumventing these circuitous methods and employing a direct guanidinylation reaction early in the synthetic sequence, it was possible to forge the linear terminal guanidine containing backbone of clavatadine A to realize a short and streamlined synthesis of this potent factor XIa inhibitor. In practice, guanidine hydrochloride is elaborated with a carefully constructed protecting array that is optimized to survive the synthetic steps to come. In the preparation of clavatadine A, direct guanidinylation of a commercially available diamine eliminated two unnecessary steps from its synthesis. Coupled with the wide variety of known guanidine protecting groups, direct guanidinylation evinces a succinct and efficient practicality inherent to methods that find a home in a synthetic chemist&#39;s toolbox.

Here, we present a protocol for direct, early stage guanidinylation that enables rapid total synthesis of aminoguanidine-containing small organic molecules. An advanced synthetic intermediate used in the synthesis of a blood coagulation factor XIa inhibitor was prepared using this protocol.

The guanidine functional group, displayed most prominently in the amino acid arginine, one of the fundamental building blocks of life, is an important ...

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain

Identification of natural ligands of chemoreceptors and structural studies aimed at elucidation of the molecular basis of the ligand specificity can be greatly facilitated by the production of milligram amounts of pure, folded ligand binding domains. Attempts to heterologously express periplasmic ligand binding domains of bacterial chemoreceptors in Escherichia coli (E. coli) often result in their targeting into inclusion bodies. Here, a method is presented for protein recovery from inclusion bodies, its refolding and purification, using the periplasmic dCACHE ligand binding domain of Campylobacter jejuni (C. jejuni) chemoreceptor Tlp3 as an example. The approach involves expression of the protein of interest with a cleavable His6-tag, isolation and urea-mediated solubilisation of inclusion bodies, protein refolding by urea depletion, and purification by means of affinity chromatography, followed by tag removal and size-exclusion chromatography. The circular dichroism spectroscopy is used to confirm the folded state of the pure protein. It has been demonstrated that this protocol is generally useful for production of milligram amounts of dCACHE periplasmic ligand binding domains of other bacterial chemoreceptors in a soluble and crystallisable form.

A procedure is presented for the refolding of the dCACHE periplasmic ligand binding domain of Campylobacter jejuni chemoreceptor Tlp3 from inclusion bodies and the purification to yield milligram quantities of protein.

Identification of natural ligands of chemoreceptors and structural studies aimed at elucidation of the molecular basis of the ligand specificity can be ...

Biochemistry

The Synthesis of [Sn10(Si(SiMe3)3)4]2- Using a Metastable Sn(I) Halide Solution Synthesized via a Co-condensation Technique

The number of well-characterized metalloid tin clusters, synthesized by applying the disproportionation of a metastable Sn(I) halide in the presence of a sterically demanding ligand, has increased in recent years. The metastable Sn(I) halide is synthesized at&#160;&#34;outer space conditions&#34; via the preparative co-condensation technique. Thereby, the subhalide is synthesized in an oven at high temperatures, around 1,300 &#176;C, and at reduced pressure by the reaction of elemental tin with hydrogen halide gas (e.g., HCl). The subhalide (e.g., SnCl) is trapped within a matrix of an inert solvent, like toluene at -196 &#176;C. Heating the solid matrix to -78 &#176;C gives a metastable solution of the subhalide. The metastable subhalide solution is highly reactive but can be stored at -78 &#176;C for several weeks. On heating the solution to room temperature, a disproportionation reaction occurs, leading to elemental tin and the corresponding dihalide. By applying bulky ligands like Si(SiMe3)3, the intermediate metalloid cluster compounds can be trapped before complete disproportionation to elemental tin. Hence, the reaction of a metastable Sn(I)Cl solution with Li-Si(SiMe3)3 gives [Sn10(Si(SiMe3)3)4]2- 1 as black crystals in high yield. 1 is formed via a complex reaction sequence including salt metathesis, disproportionation, and degradation of larger clusters. Further, 1 can be analyzed by various methods like NMR or single crystal X-ray structure analysis.

The Synthesis of [Sn10SiSiMe334]2- Using a Metastable SnI Halide Solution Synthesized via a Co-condensation Technique

The disproportionation reaction of a metastable Sn(I) chloride solution, obtained via the preparative co-condensation technique, is used for the synthesis of a metalloid tin cluster compound.

The number of well-characterized metalloid tin clusters, synthesized by applying the disproportionation of a metastable Sn(I) halide in the presence of a ...

A Dual-Functional Electroactive Filter Towards Simultaneously Sb(III) Oxidation and Sequestration

We have designed a facile method to synthesize a dual-functional electrochemical filter consisting of two 1-D materials: titanate nanowires and carbon nanotubes. The hybrid titanate-CNT filter was prepared by a sonication coupled with a post-filtration route. Due to the synergistic effects of the increased number of exposed sorption sites, electrochemical reactivity, small pore size of the titanate-CNT network coupled with a flow-through design, simultaneous Sb(III) oxidation and sequestration can be readily achieved. Atomic fluorescence spectrometer technology demonstrated that the applied electrical field accelerates the Sb(III) conversion rate and the as-obtained Sb(V) were adsorbed effectively by the titanate nanowires due to their Sb specificity. This protocol provides a practical solution for the removal of highly toxic Sb(III) and other similar heavy metal ions.

A Dual-Functional Electroactive Filter Towards Simultaneously SbIII Oxidation and Sequestration

A protocol for the rational design of a dual-functional electroactive filter consisting of carbon nanotubes and titanate nanowires is reported and their environmental applications towards Sb(III) oxidation and sequestration is presented.

We have designed a facile method to synthesize a dual-functional electrochemical filter consisting of two 1-D materials: titanate nanowires and carbon ...

Environment

Synthesis of a Thiol Building Block for the Crystallization of a Semiconducting Gyroidal Metal-sulfur Framework

We present a method for preparing thioester molecules as the masked form of the thiol linkers and their utilization for accessing a semiconducting and porous metal-dithiolene network in the highly ordered single crystalline state. Unlike the highly reactive free-standing thiols, which tend to decompose and complicate the crystallization of metal-thiolate open frameworks, the thioester reacts in situ to provide the thiol species, serving to mitigate the reaction between the mercaptan units and the metal centers, and to improve crystallization consequently. Specifically, the thioester was synthesized in a one-pot procedure: an aromatic bromide (hexabromotriphenylene) reacted with excess sodium thiomethoxide under vigorous conditions to first form the thioether intermediate product. The thioether was then demethylated by the excess thiomethoxide to provide the thiolate anion that was acylated to form the thioester product. The thioester was conveniently purified by standard column chromatography, and then used directly in the framework synthesis, wherein NaOH and ethylenediamine serve to revert in situ the thioester to the thiol linker for assembling the single-crystalline Pb(II)-dithiolene network. Compared with other methods for thiol synthesis (e.g., by cleaving alkyl thioether using sodium metal and liquid ammonia), the thioester synthesis here uses simple conditions and economical reagents. Moreover, the thioester product is stable and can be conveniently handled and stored. More importantly, in contrast to the generic difficulty in accessing crystalline metal-thiolate open frameworks, we demonstrate that using the thioester for in situ formation of the thiol linker greatly improves the crystallinity of the solid-state product. We intend to encourage broader research efforts on the technologically important metal-sulfur frameworks by disclosing the synthetic protocol for the thioester as well as the crystalline framework solid.

Here, we present a one-pot, transition-metal-free synthesis of thiols and thioesters from aromatic halides and sodium thiomethoxide, followed by the preparation of single crystals of a metal-dithiolene network using thiol species generated in situ from the more stable and tractable thioester.

We present a method for preparing thioester molecules as the masked form of the thiol linkers and their utilization for accessing a semiconducting and ...

Separation of Aldehydes and Reactive Ketones from Mixtures Using a Bisulfite Extraction Protocol

The purification of organic compounds is an essential component of routine synthetic operations. The ability to remove contaminants into an aqueous layer by generating a charged structure provides an opportunity to use extraction as a simple purification technique. By combining the use of a miscible organic solvent with saturated sodium bisulfite, aldehydes and reactive ketones can be successfully transformed into charged bisulfite adducts that can then be separated from other organic components of a mixture by the introduction of an immiscible organic layer. Here, we describe a simple protocol for the removal of aldehydes, including sterically-hindered neopentyl aldehydes and some ketones, from chemical mixtures. Ketones can be separated if they are sterically unhindered cyclic or methyl ketones. For aliphatic aldehydes and ketones, dimethylformamide is used as the miscible solvent to improve removal rates. The bisulfite addition reaction can be reversed by basification of the aqueous layer, allowing for the re-isolation of the reactive carbonyl component of a mixture.

Here, we present a protocol to remove aldehydes and reactive ketones from mixtures by a liquid-liquid extraction protocol directly with saturated sodium bisulfite in a miscible solvent. This combined protocol is rapid and facile to perform. The aldehyde or ketone can be re-isolated by the basification of the aqueous layer.

The purification of organic compounds is an essential component of routine synthetic operations. The ability to remove contaminants into an aqueous layer ...

Easy Access to Aliphatic Sulfonamides using Sulfamoyl Chlorides Under Visible Light Activation

Sulfonamides are prevalent motifs in marketed drugs and natural products. Their synthesis represents a great interest to the pharmaceutical industry, due to their unique biological properties. Recently, several methods for the synthesis of aryl sulfonamides have been developed, but little effort has focused on developing one-step methodologies to access sulfonamides flanked by two alkyl groups. This protocol describes a practical and facile method for the net hydrosulfamoylation of electron-deficient alkenes using sulfamoyl chlorides as radical precursors under blue-light activation. This practical and cost-effective methodology is performed in the presence of the metal-free photocatalyst Eosin Y and uses light as a clean and traceless energy source. The procedure is scalable, displays a broad functional group tolerance, and can be applied for late-stage functionalization. All reagents used in this protocol are commercially available. Simple reaction set-up, the absence of work-up and easy purification, demonstrate the convenience of this protocol. The reaction is best applied to electron-deficient alkenes.

Presented here is a protocol for the easy synthesis of aliphatic sulfonamides using sulfamoyl chlorides, (TMS)3SiH and Eosin Y under blue-light irradiation.

Sulfonamides are prevalent motifs in marketed drugs and natural products. Their synthesis represents a great interest to the pharmaceutical industry, due ...

Crystallization of ABC Transporter Membrane Proteins in Lipidic Bicelles

ABCG5/G8 Crystallization in a Lipidic Bicelle Environment for X-Ray Crystallography

ATP-binding cassette (ABC) transporters constitute lipid-embedded membrane proteins. Extracting these membrane proteins from the lipid bilayer to an aqueous environment is typically achieved by employing detergents. These detergents disintegrate the lipid bilayer and solubilize the proteins. The intrinsic habitat of membrane proteins within the lipid bilayer poses a challenge in maintaining their stability and uniformity in solution for structural characterization. Bicelles, which comprise a blend of long and short-chain phospholipids and detergents, replicate the natural lipid structure. The utilization of lipid bicelles and detergents serves as a suitable model system for obtaining high-quality diffraction crystals, specifically to determine the high-resolution structure of membrane proteins. Through these synthetic microenvironments, membrane proteins preserve their native conformation and functionality, facilitating the formation of three-dimensional crystals. In this approach, the detergent-solubilized heterodimeric ABCG5/G8 was reintegrated into DMPC/CHAPSO bicelles, supplemented with cholesterol. This setup was employed in the vapor diffusion experimental procedure for protein crystallization.

Author Spotlight: A Bicelle Crystallization Setup for ABC Transporter Membrane Proteins to Advance Drug Development 

This protocol describes a setup for the crystallization of the sterol transporter ABCG5/G8. ABCG5/G8 is reconstituted into bicelles for hanging-drop crystallization. The protocol does not require specialized materials or substrates, making it accessible and easy to adapt in any laboratory for determining the protein structure through X-ray crystallography.

ATP-binding cassette (ABC) transporters constitute lipid-embedded membrane proteins. Extracting these membrane proteins from the lipid bilayer to an ...

Synthesis of Nine-atom Deltahedral Zintl Ions of Germanium and their Functionalization with Organic Groups

Although the first studies of Zintl ions date between the late 1890's and early 1930's they were not structurally characterized until many years later.1,2 Their redox chemistry is even younger, just about ten years old, but despite this short history these deltahedral clusters ions E9n- (E = Si, Ge, Sn, Pb; n = 2, 3, 4) have already shown interesting and diverse reactivity and have been at the forefront of rapidly developing and exciting new chemistry.3-6 Notable milestones are the oxidative coupling of Ge94- clusters to oligomers and infinite chains,7-19 their metallation,14-16,20-25 capping by transition-metal organometallic fragments,26-34 insertion of a transition-metal atom at the center of the cluster which is sometimes combined with capping and oligomerization,35-47 addition of main-group organometallic fragments as exo-bonded substituents,48-50 and functionalization with various organic residues by reactions with organic halides and alkynes.51-58 
This latter development of attaching organic fragments directly to the clusters has opened up a new field, namely organo-Zintl chemistry, that is potentially fertile for further synthetic explorations, and it is the step-by-step procedure for the synthesis of germanium-divinyl clusters described herein. The initial steps outline the synthesis of an intermetallic precursor of K4Ge9 from which the Ge94- clusters are extracted later in solution. This involves fused-silica glass blowing, arc-welding of niobium containers, and handling of highly air-sensitive materials in a glove box. The air-sensitive K4Ge9 is then dissolved in ethylenediamine in the box and then alkenylated by a reaction with Me3SiC&equiv;CSiMe3. The reaction is followed by electrospray mass spectrometry while the resulting solution is used for obtaining single crystals containing the functionalized clusters [H2C=CH-Ge9-CH=CH2]2-. For this purpose the solution is centrifuged, filtered, and carefully layered with a toluene solution of 18-crown-6. Left undisturbed for a few days, the so-layered solutions produced orange crystalline blocks of [K(18-crown-6)]2[Ge9(HCCH2)2]&bull;en which were characterized by single-crystal X-ray diffraction. 
The process highlights standard reaction techniques, work-up, and analysis towards functionalized deltahedral Zintl clusters. It is hoped that it will help towards further development and understanding of these compounds in the community at large.

We present the high-temperature synthesis of intermetallic precursors K4Ge9, their dissolution in ethylenediamine to form Ge94- deltahedral Zintl ions, and the reaction of the clusters with alkynes to form organo-Zintl ions. The latter are characterized by electrospray mass spectrometry in solutions and by single-crystal X-ray diffraction in the solid state.

Although the first studies of Zintl ions date between the late 1890's and early 1930's they were not structurally characterized until many years later.1,2 ...

Sulfate Separation by Selective Crystallization with a Bis-iminoguanidinium Ligand

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