This work was conducted while Bian, Chen, and Wang were visiting Ph.D. students at the US Forest Service, Forest Products Laboratory (FPL), Madison, WI, and on official government time of Zhu. This work was partially supported by the USDA Agriculture and Food Research Initiative (AFRI) Competitive Grant (No. 2011-67009-20056), the Chinese State Forestry Administration (Project No. 2015-4-54), the National Natural Science Foundation of China (Project No. 31470599), Guangzhou Elite Project of China, and China Scholarship Fund. Funding from these programs made the visiting appointments of Bian, Chen, and Wang at FPL possible.

NOTE: Bleached eucalyptus kraft pulp (BEP) and unbleached mixed hardwood kraft pulp (UMHP) fibers from commercial sources were used as feedstock for producing CNC and CNF. Commercial maleic acids purchased were used for hydrolysis. Hydrolysis conditions were acid concentrations of 60 wt% at 100 &#176;C for 60 min and 120 &#176;C (no boiling at atmospheric pressure) for 120 min, for BEP and UMHP, respectively.
1. Preparation of Concentrated Dicarboxylic Acid Solution
<ol>
	<li>Heat 40 mL deionized (DI) water in a multiple-neck flask in a liquid glycerol bath on a heating plate to approximately 85 &#176;C.</li>
	<li>Add 60 g anhydrous maleic acid into the flask to make a 60 wt% solution with magnetic stirring. Using the densities of the acid solutions reported previously 15, calculate the required amount of water and acid for making the acid solution at the specified mass concentration.</li>
	<li>Heat the solution to the desired hydrolysis temperature of 100 or 120 &#176;C (no boiling due to high dicarboxylic acid concentrations).</li>
</ol>
2. Hydrolysis Reaction
<ol>
	<li>Once the acid solution is at temperature, add 10 g oven dried (OD) of BEP or UMHP fibers into 80 mL dicarboxylic acid solution (1.1) with continuous stirring.
	<ol>
		<li>Take an aliquot of the acid hydrolysate (approximately 2 mL) at the end of the predetermined reaction time of 60 min before terminating hydrolysis by adding 160 mL of 80 &#176;C DI water.</li>
	</ol>
	</li>
	<li>Dilute 0.5 mL of the sampled hydrolysate for sugar and acid concentration analyses. 15 Observe the remaining hydrolysate sample for crystallization taking place while cooing down to room temperature.</li>
	<li>Separate the hydrolysate from the hydrolyzed pulp by vacuum filtration using a filter paper in a Buchner funnel. 
	NOTE: This separation needs to occur rather quickly before the temperature drops and the acid begins to crystallize out of solution. Due to high ionic strength from the acid solution, the CNC&#39;s generated in hydrolysis agglomerate and stay with the FCSR residuals. Approximately 80-90% of the acid will be removed with the filtrate.</li>
</ol>
3. CNC Separation
<ol>
	<li>Wash the filtered solids from section 2 using DI water and dilute to approximately 1% total solids with DI water. Centrifuge the filtrate at 11,960 x g for 10 min.</li>
	<li>Decant off the supernatant. Repeat the washing and filtration process using fresh DI water until the supernatant is turbid. The turbidity indicates that the ionic strength of the solution has fallen enough for the CNC to disperse and start to become colloidal.</li>
	<li>Mix the turbid supernatant with the settled hydrolyzed pulp (2.3). Dialyze the mixture in a dialysis bag (MWCO 14 kDa) using DI water until the conductivity of the liquid approaches that of DI water. Measure the conductivity using a conductance meter.</li>
	<li>Centrifuge the dialyzed sample at 3,500 x g for 10 min to obtain a CNC dispersion in the aqueous phase. Retain the precipitate phase, i.e., FCSR, for CNF production.</li>
	<li>Determine CNC yield from the measured amount of CNC in the dispersion using a COD method described previously 3, 18.</li>
</ol>
4. CNF Production
<ol>
	<li>Determine the yield of precipitated FCSR by gravimetric measurements after separating the CNC dispersion. Dry the FCSR at 105 &#176;C and measure the oven dry weight of the FCSR relative to the initial oven dry weight of BEP or UMHP fibers used.</li>
	<li>Mechanically fibrillate the FCSR at fiber suspension of 0.5% by consecutively passing the suspension 3 times through a 200 &#181;m orifice chamber followed by 2 times through an 87 &#181;m orifice chamber, all at 100 MPa.</li>
</ol>
5. Atomic Force Microscopy (AFM) Imaging
<ol>
	<li>Sonicate approximately 0.01 wt% CNC or CNF suspensions for 2 min. Deposit a drop of the dispersed suspension on a mica substrate. Air-dry the deposited suspension at ambient temperature.</li>
	<li>Take AFM images of the air-dried CNCs and CNFs in vibrating tapping mode using manufacturer&#39;s protocol. Analyze the AFM images of approximately 100 individual CNCs or CNFs using commercial software to obtain diameter and length distributions.</li>
</ol>
6. Fourier Transform Inferred (FTIR) Measurements
<ol>
	<li>Use a commercial FTIR spectrophotometer with a universal attenuated-total-reflection (ATR) probe to analyze the resultant CNC and CNF samples along with the original BEP and UMHP fibers to identify ester groups.</li>
	<li>Record the absorption spectra of the samples in a wavelength range between 450 - 4,000 cm-1 with a resolution of 4 cm-1 and 4 scans for each sample.</li>
</ol>
7. Conductometric Titration
<ol>
	<li>Use conductometric titration to quantify the carboxyl group contents of samples resulted from esterification by the dicarboxylic acid.</li>
	<li>Add CNC or CNF suspension with 50 mg (OD) of CNCs or CNFs into 120 mL of 1 mM NaCl solution. Titrate the mixture by adding approximately 0.2 mL of 2 mM NaOH solution at 30 s intervals.</li>
	<li>Measure the conductivity using a conductance meter. Find the inflection point (the lowest point) on the conductivity curve during the course of adding NaOH.</li>
	<li>Calculate the amount of carboxyl groups (mmol/L) based on the consumed NaOH relative to the inflection point using the following equation in which c is the concentration of NaOH solution (mol/L), v is the volume of added NaOH solution (mL), m is the mass of CNCs or CNFs in OD weight (g). 
	<img alt="Equation" src="/files/ftp_upload/55079/55079eq1.jpg" /></li>
</ol>
8. CNC and CNF Thermal Stability Determination
<ol>
	<li>Conduct thermal degradation measurements of the CNC and CNF samples by thermogravimetric analysis (TGA).</li>
	<li>Purge the furnace using a high purity nitrogen flow at 20 mL/min to prevent any unwanted oxidative decomposition. Dry the samples at 50 &#176;C for 4 h before testing. Use a sample size of 5 mg in dry weight.</li>
	<li>Record the weight of the sample as the furnace temperature is increased from ambient to 600 &#176;C at a heating rate of 10 &#176;C/min.</li>
	<li>Normalize the measured weight loss by the initial weight.</li>
	<li>Conduct separate thermal stability tests of CNC and CNF samples in an oven at 105 &#176;C. Record the color change of the samples after 4 and 24 h by conventional photography.</li>
</ol>
9. X-ray Diffraction
<ol>
	<li>Press freeze-dried CNC or CNF samples at 180 MPa to make pellets as described previously. 5 Conduct wide angle X-ray diffraction measurements of the pellet using Cu-K&#945; radiation on an X-ray diffractometer in the 2&#952; range of 10 - 38&#176; in steps of 0.02&#176;.</li>
	<li>Calculate the crystallinity index (CrI) of a pellet using the Segal method 19 (without base line subtraction).</li>
</ol>

<ol>
	<li>Giese, M., Blusch, L. K., Khan, M. K., MacLachlan, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+Materials+from+Cellulose-Derived+Liquid-Crystal+Templates.">Functional Materials from Cellulose-Derived Liquid-Crystal Templates.</a> Angew Chem Int Ed. 54 (10), 2888-2910 (2015).</li><li>Zhu, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wood-Derived+Materials+for+Green+Electronics,+Biological+Devices,+and+Energy+Applications+.">Wood-Derived Materials for Green Electronics, Biological Devices, and Energy Applications .</a> Chem. Rev. , (2016).</li><li>Wang, Q. Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Approaching+zero+cellulose+loss+in+cellulose+nanocrystal+(CNC)+production:+recovery+and+characterization+of+cellulosic+solid+residues+(CSR)+and+CND.">Approaching zero cellulose loss in cellulose nanocrystal (CNC) production: recovery and characterization of cellulosic solid residues (CSR) and CND.</a> Cellulose. 19 (6), 2033-2047 (2012).</li><li>Hamad, W. Y., Hu, T. Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure-process-yield+interrelations+in+nanocrystalline+cellulose+extraction.">Structure-process-yield interrelations in nanocrystalline cellulose extraction.</a> Can J Chem Eng. 88 (3), 392-402 (2010).</li><li>Chen, L. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tailoring+the+yield+and+characteristics+of+wood+cellulose+nanocrystals+(CNC)+using+concentrated+acid+hydrolysis.">Tailoring the yield and characteristics of wood cellulose nanocrystals (CNC) using concentrated acid hydrolysis.</a> Cellulose. 22 (3), 1753-1762 (2015).</li><li>Mukherjee, S. M., Woods, H. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=X-ray+and+electron+microscope+studies+of+the+degradation+of+cellulose+by+sulphuric+acid.">X-ray and electron microscope studies of the degradation of cellulose by sulphuric acid.</a> Biochim Biophys Acta. 10 (4), 499-511 (1953).</li><li>Camarero Espinosa, S., Kuhnt, T., Foster, E. J., Weder, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+thermally+stable+cellulose+nanocrystals+by+phosphoric+acid+hydrolysis.">Isolation of thermally stable cellulose nanocrystals by phosphoric acid hydrolysis.</a> Biomacromolecules. 14 (4), 1223-1230 (2013).</li><li>Yu, H. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facile+extraction+of+thermally+stable+cellulose+nanocrystals+with+a+high+yield+of+93%+through+hydrochloric+acid+hydrolysis+under+hydrothermal+conditions.">Facile extraction of thermally stable cellulose nanocrystals with a high yield of 93% through hydrochloric acid hydrolysis under hydrothermal conditions.</a> J Mater Chem, A. 1 (12), 3938-3944 (2013).</li><li>Leung, A. C. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characteristics+and+properties+of+carboxylated+cellulose+nanocrystals+prepared+from+a+novel+one-step+procedure.">Characteristics and properties of carboxylated cellulose nanocrystals prepared from a novel one-step procedure.</a> Small. 7 (3), 302-305 (2011).</li><li>Saito, T., Isogai, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TEMPO-mediated+oxidation+of+native+cellulose.+The+effect+of+oxidation+conditions+on+chemical+and+crystal+structures+of+the+water-insoluble+fractions.">TEMPO-mediated oxidation of native cellulose. The effect of oxidation conditions on chemical and crystal structures of the water-insoluble fractions.</a> Biomacromolecules. 5 (5), 1983-1989 (2004).</li><li>Yang, H., Chen, D. Z., van de Ven, T. G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+characterization+of+sterically+stabilized+nanocrystalline+cellulose+obtained+by+periodate+oxidation+of+cellulose+fibers.">Preparation and characterization of sterically stabilized nanocrystalline cellulose obtained by periodate oxidation of cellulose fibers.</a> Cellulose. 22 (3), 1743-1752 (2015).</li><li>Huang, Y. B., Fu, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydrolysis+of+cellulose+to+glucose+by+solid+acid+catalysts.">Hydrolysis of cellulose to glucose by solid acid catalysts.</a> Green Chem. 15 (5), 1095-1111 (2013).</li><li>Shimizu, K. I., Satsuma, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+a+rational+control+of+solid+acid+catalysis+for+green+synthesis+and+biomass+conversion.">Toward a rational control of solid acid catalysis for green synthesis and biomass conversion.</a> Energy &amp; Environ Sci. 4 (9), 3140-3153 (2011).</li><li>Wang, Q. Q., Zhu, J. Y., Considine, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strong+and+optically+transparent+films+prepared+using+cellulosic+solid+residue+(CSR)+recovered+from+cellulose+nanocrystals+(CNC)+production+waste+stream.">Strong and optically transparent films prepared using cellulosic solid residue (CSR) recovered from cellulose nanocrystals (CNC) production waste stream.</a> ACS Appl Mater Interfaces. 5 (7), 2527-2534 (2013).</li><li>Chen, L. H., Zhu, J. Y., Baez, C., Kitin, P., Elder, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Highly+thermal-stable+and+functional+cellulose+nanocrystals+and+nanofibrils+produced+using+fully+recyclable+organic+acids.">Highly thermal-stable and functional cellulose nanocrystals and nanofibrils produced using fully recyclable organic acids.</a> Green Chem. 18, 3835-3843 (2016).</li><li>Fischer, E., Speier, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Darstellungder+der+Ester.">Darstellungder der Ester.</a> Chemische Berichte. 28 (3), 3252-3258 (1895).</li><li>Allen, T. C., Cuculo, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellulose+derivatives+containing+carboxylic+acid+groups.">Cellulose derivatives containing carboxylic acid groups.</a> J Polym Sci: Macromol Rev. 7 (1), 189-262 (1973).</li><li>Wang, Q. Q., Zhao, X. B., Zhu, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Kinetics+of+strong+acid+hydrolysis+of+a+bleached+kraft+pulp+for+producing+cellulose+nanocrystals+(CNCs).">Kinetics of strong acid hydrolysis of a bleached kraft pulp for producing cellulose nanocrystals (CNCs).</a> Ind Eng Chem Res. 53 (27), 11007-11014 (2014).</li><li>Segal, L., Creely, J. J., Martin, A. E., Conrad, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+empirical+method+for+estimating+the+degree+of+crystallinity+of+native+cellulose+using+the+X-ray+diffractometer.">An empirical method for estimating the degree of crystallinity of native cellulose using the X-ray diffractometer.</a> Text Res J. 29 (10), 786-794 (1959).</li></ol>

Chen and Zhu are co-inventors of a US patent application using dicarboxylic acids for CNC and CNF production.

The thicker CNC diameters of the CNC samples from maleic acid hydrolysis resulted in a moderate average aspect ratio 7.24 and 8.53, for the CNCs from BEP and UMHP, respectively, despite their long lengths as discussed above. The CNFs had a longer length and a thinner diameter, which resulted in a large aspect ratio of 13.9 and 19.0, for the CNCs from BEP and UMHP, respectively, both greater than their respective CNCs. It is possible to use severe mechanical fibrillation to reduce CNF diameter to improve the aspect ratio ...

Sustainable economic development requires not only using feedstocks that are renewable and biodegradable but also uses green and environmental friendly manufacturing technologies to produce a variety of bioproducts and biochemicals from these renewable feedstocks. Cellulose nanomaterials, such as cellulose nanocrystals (CNC) and cellulose nanofibrils (CNF), produced from renewable lignocelluloses are biodegradable and have unique mechanical and optical properties suitable for developing a range of bioproducts 1, 2. Unfortunately, existing technologies for producing cellulose nanomaterials are either energy intensive when using pure mechanical fibrillation or environmentally unsustainable due to non-recycling or insufficient recycling of processing chemicals, such as when using the concentrated mineral acid hydrolysis process 3-8 or oxidation methods 9-11. Furthermore, oxidation methods may also produce environmentally toxic compounds by reacting with lignocelluloses. Therefore, developing green manufacturing technologies for producing cellulose nanomaterials is critically important to make full use of the abundant and renewable material - lignocelluloses.
Using acid hydrolysis to dissolve hemicellulose and depolymerize cellulose is an effective approach for producing cellulose nanomaterials. Solid acids have been used for sugar production from cellulose with the advantage of easing acid recovery 12, 13. Previous studies using concentrated mineral acids indicated that a lower acid concentration improved CNC yield and crystallinity 3, 5. This suggests that a strong acid may damage cellulose crystals while a milder acid hydrolysis might improve the properties and yield of cellulose nanomaterials through the approach of integrated production and CNC with CNF 3, 14. Here we document a method using concentrated solid dicarboxylic acids hydrolysis to produce CNC along with CNF 15. These dicarboxylic acids have low solubility at low or ambient temperatures, and therefore they can be easily recovered through the mature crystallization technology. They also have good solubility at elevated temperatures which facilitates concentrated acid hydrolysis without boiling or using pressure vessels. Since these acids also have a higher pKa than typical mineral acids used for CNC production, their use results in good CNC crystallinity, and despite lower CNC yields, with a substantial amount of fibrous cellulosic solid residue (FCSR or partially hydrolyzed fibers) remaining due to incomplete cellulose depolymerization. The FCSR can be used to produce CNF through subsequent mechanical fibrillation using low energy inputs. Therefore, cellulose loss to sugars is minimal as compared to using mineral acids.
It is well known that carboxylic acids can esterify cellulose through Fisher-Speier esterification 16. Applying dicarboxylic acids to cellulose can result in semi-acid un-crosslinked esters 17 (or carboxylation), to produce carboxylated CNC and CNF as we demonstrated 15 previously. The method documented here can produce carboxylated and thermally stable CNF and CNC that is also highly crystalline from either bleached or unbleached pulps while having relatively simple and high chemical recovery and using low energy inputs.

<table border="0" cellpadding="0" cellspacing="0" width="1222">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:349px;">Bleached eucalypus pulp&#160;</td>
			<td style="width:327px;">Aracruz Cellulose</td>
			<td style="width:248px;"></td>
			<td style="width:299px;"></td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:349px;">Unbleached mixed hardwood kraft pulp&#160;</td>
			<td>International Paper&#160;</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="26">
			<td height="26" style="height:27px;">Maleic acid</td>
			<td>Sigma-Aldrich</td>
			<td>M0375-1KG/CAS110-16-7</td>
			<td style="width:299px;">Powder; assay: 99.0%(HPLC)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glycerol</td>
			<td>Sigma-Aldrich</td>
			<td>G5516-4L/CAS56-81-5</td>
			<td></td>
		</tr>
		<tr height="37">
			<td height="37" style="height:38px;">Sodium hydroxide</td>
			<td>Fisher Scientific</td>
			<td style="width:248px;">S318-500/CAS1310-73-2, 497-19-8</td>
			<td>Certified ACS</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:25px;">Sodium chloride</td>
			<td>Mallinckrodt</td>
			<td>7581-12/CAS7647-14-5</td>
			<td>Crystal,AR</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:42px;">Cupriethylenediamine solution</td>
			<td>GFS Chemicals</td>
			<td>E32103-1L/CAS14552-35-3</td>
			<td style="width:299px;">1M, for determination of solution viscosity of pulps</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:25px;">Acetone</td>
			<td>Fisher Scientific</td>
			<td>A18-500/CAS67-64-1</td>
			<td>Certified ACS</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Accu-TestTM Vials for COD Testing</td>
			<td>Bioscience,Inc.</td>
			<td>01-215-28</td>
			<td style="width:299px;">COD testing for 20 to 900mg/L standard range concentration</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Heating plate</td>
			<td>IKA</td>
			<td></td>
			<td>Mode: C-MAD HS7 digital</td>
		</tr>
		<tr height="29">
			<td height="29" style="height:30px;">Magnetic stir bar</td>
			<td>ACE Glass</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="29">
			<td height="29" style="height:30px;">Pyrex three-neck round-bottom flask</td>
			<td>Sigma-Aldrich</td>
			<td>CLS4965B500-1EA</td>
			<td></td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;">Dialysis tubing cellulose membrane</td>
			<td>Sigma-Aldrich</td>
			<td>D9402-100FT</td>
			<td style="width:299px;">Typical molecular weight cut-off = 14000</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:23px;">Disposable aluminum dishes</td>
			<td>Sigma-Aldrich</td>
			<td>Z154857-1PAK</td>
			<td>Circles, 60mm</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Disintegrator</td>
			<td>Testing Machines Inc.(TMI)</td>
			<td></td>
			<td style="width:299px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Microfluidizer</td>
			<td>Microfluidics Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sonicator</td>
			<td>Qsonica LLC.</td>
			<td></td>
			<td>Mode: 3510R-MT, 50-60 Hz,180 W</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Zeta potential analyzer</td>
			<td>Brookhaven Instruments Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">FTIR</td>
			<td>PerkinElmer</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Conductometric titrator</td>
			<td>Yellow Springs Instrument (YSI)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">TGA analyzer</td>
			<td>PerkinElmer</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">X-ray diffractometer</td>
			<td>Bruker Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">AFM imging&#160;</td>
			<td>AFM Workshop</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SEM imaging</td>
			<td>Carl Zeiss</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

green and low-cost production of thermally stable and carboxylated cellulose nanocrystals and nanofibrils using highly recyclable dicarboxylic acids

Here we demonstrate potentially low cost and green productions of high thermally stable and carboxylated cellulose nanocrystals (CNCs) and nanofibrils (CNF) from bleached eucalyptus pulp (BEP) and unbleached mixed hardwood kraft pulp (UMHP) fibers using highly recyclable dicarboxylic solid acids. Typical operating conditions were acid concentrations of 50 - 70 wt% at 100 &#176;C for 60 min and 120 &#176;C (no boiling at atmospheric pressure) for 120 min, for BEP and UMHP, respectively. The resultant CNCs have a higher thermal degradation temperature than their corresponding feed fibers and carboxylic acid group content from 0.2 - 0.4 mmol/g. The low strength (high pKa of 1.0 - 3.0) of organic acids also resulted in CNCs with both longer lengths of approximately 239 - 336 nm and higher crystallinity than CNCs produced using mineral acids. Cellulose loss to sugar was minimal. Fibrous cellulosic solid residue (FCSR) from the dicarboxylic acid hydrolysis was used to produce carboxylated CNFs through subsequent mechanical fibrillation with low energy input.

Typical AFM images of the CNC and CNF from BEP and UMHP along with corresponding SEM images of the feed acid hydrolyzed fibers are shown in Figures 1 and 2. The images clearly show the substantial reductions in fiber length by acid hydrolysis with minimal change in fiber diameters (comparing Figure 1a with 1b, and 2a with 2b). The shortened fiber length was also reflected by the measured cellulose degree of polymerizatio...

Watch this Scientific Journal Video about Green and Low-cost Production of Thermally Stable and Carboxylated Cellulose Nanocrystals and Nanofibrils Using Highly Recyclable Dicarboxylic Acids at JoVE.c

Green and Low-cost Production of Thermally Stable and Carboxylated Cellulose Nanocrystals and Nanofibrils Using Highly Recyclable Dicarboxylic Acids

Here we demonstrate a novel method for green and sustainable productions of highly thermally stable and carboxylated cellulose nanocrystals (CNC) and nanofibrils (CNF) using highly recyclable solid dicarboxylic acids.

Here we demonstrate potentially low cost and green productions of high thermally stable and carboxylated cellulose nanocrystals (CNCs) and nanofibrils ...

green-low-cost-production-thermally-stable-carboxylated-cellulose

Research

JoVE Journal

Chemistry

11.8K Views.  Nanjing Forestry University. Here we demonstrate a novel method for green and sustainable productions of highly thermally stable and carboxylated cellulose nanocrystals (CNC) and nanofibrils (CNF) using highly recyclable solid dicarboxylic acids.

Video: Green and Low-cost Production of Thermally Stable and Carboxylated Cellulose Nanocrystals and Nanofibrils Using Highly Recyclable Dicarboxylic Acids

Seeded Synthesis of CdSe/CdS Rod and Tetrapod Nanocrystals

We demonstrate a method for the synthesis of multicomponent nanostructures consisting of CdS and CdSe with rod and tetrapod morphologies. A seeded synthesis strategy is used in which spherical seeds of CdSe are prepared first using a hot-injection technique. By controlling the crystal structure of the seed to be either wurtzite or zinc-blende, the subsequent hot-injection growth of CdS off of the seed results in either a rod-shaped or tetrapod-shaped nanocrystal, respectively. The phase and morphology of the synthesized nanocrystals are confirmed using X-ray diffraction and transmission electron microscopy, demonstrating that the nanocrystals are phase-pure and have a consistent morphology. The extinction coefficient and quantum yield of the synthesized nanocrystals are calculated using UV-Vis absorption spectroscopy and photoluminescence spectroscopy. The rods and tetrapods exhibit extinction coefficients and quantum yields that are higher than that of the bare seeds. This synthesis demonstrates the precise arrangement of materials that can be achieved at the nanoscale by using a seeded synthetic approach.

A protocol for the seeded synthesis of rod-shaped and tetrapod-shaped multicomponent nanostructures consisting of CdS and CdSe is presented.

We demonstrate a method for the synthesis of multicomponent nanostructures consisting of CdS and CdSe with rod and tetrapod morphologies. A seeded ...

Qualitative Identification of Carboxylic Acids, Boronic Acids, and Amines Using Cruciform Fluorophores

Molecular cruciforms are X-shaped systems in which two conjugation axes intersect at a central core. If one axis of these molecules is substituted with electron-donors, and the other with electron-acceptors, cruciforms&#39; HOMO will localize along the electron-rich and LUMO along the electron-poor axis. This spatial isolation of cruciforms&#39; frontier molecular orbitals (FMOs) is essential to their use as sensors, since analyte binding to the cruciform invariably changes its HOMO-LUMO gap and the associated optical properties. Using this principle, Bunz and Miljani&#263; groups developed 1,4-distyryl-2,5-bis(arylethynyl)benzene and benzobisoxazole cruciforms, respectively, which act as fluorescent sensors for metal ions, carboxylic acids, boronic acids, phenols, amines, and anions. The emission colors observed when these cruciform are mixed with analytes are highly sensitive to the details of analyte&#39;s structure and - because of cruciforms&#39; charge-separated excited states - to the solvent in which emission is observed. Structurally closely related species can be qualitatively distinguished within several analyte classes: (a) carboxylic acids; (b) boronic acids, and (c) metals. Using a hybrid sensing system composed from benzobisoxazole cruciforms and boronic acid additives, we were also able to discern among structurally similar: (d) small organic and inorganic anions, (e) amines, and (f) phenols. The method used for this qualitative distinction is exceedingly simple. Dilute solutions (typically 10-6 M) of cruciforms in several off-the-shelf solvents are placed in UV/Vis vials. Then, analytes of interest are added, either directly as solids or in concentrated solution. Fluorescence changes occur virtually instantaneously and can be recorded through standard digital photography using a semi-professional digital camera in a dark room. With minimal graphic manipulation, representative cut-outs of emission color photographs can be arranged into panels which permit quick naked-eye distinction among analytes. For quantification purposes, Red/Green/Blue values can be extracted from these photographs and the obtained numeric data can be statistically processed.

Cross-conjugated cruciform fluorophores based on 1,4-distyryl-2,5-bis(arylethynyl)benzene and benzobisoxazole nuclei can be used to qualitatively identify diverse Lewis acidic and Lewis basic analytes. This method relies on the differences in emission colors of the cruciforms that are observed upon analyte addition. Structurally closely related species can be distinguished from each other.

Molecular cruciforms are X-shaped systems in which two conjugation axes intersect at a central core. If one axis of these molecules is substituted with ...

Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

A common screen for plant antimicrobial compounds consists of separating plant extracts by paper or thin-layer chromatography (PC or TLC), exposing the chromatograms to microbial suspensions (e.g. fungi or bacteria in broth or agar), allowing time for the microbes to grow in a humid environment, and visualizing zones with no microbial growth.&#160;The effectiveness of this screening method, known as bioautography, depends on both the quality of the chromatographic separation and the care taken with microbial culture conditions.&#160;This paper describes standard protocols for TLC and contact bioautography&#160;with a novel application to amino acid-fermenting bacteria. The extract is separated on flexible (aluminum-backed) silica TLC plates, and bands are visualized under ultraviolet (UV) light.&#160;Zones are cut out and incubated face down onto agar inoculated with the test microorganism.&#160;Inhibitory bands are visualized by staining the agar plates with tetrazolium red.&#160;The method is applied to the separation of red clover (Trifolium pratense cv. Kenland) phenolic compounds and their screening for activity against Clostridium sticklandii, a hyper ammonia-producing bacterium (HAB) that is native to the bovine rumen.&#160;The TLC methods apply to many types of plant extracts&#160;and other bacterial species (aerobic or anaerobic), as well as fungi, can be used as test organisms if culture conditions are modified to fit the growth requirements of the species.

Thin-layer Chromatographic TLC Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

Methods are described for thin-layer chromatographic (TLC) separation of plant extracts and contact bioautography to identify antibacterial metabolites. The methods are applied to the screening of red clover phenolic compounds inhibiting hyper&#160;ammonia-producing bacteria (HAB) native to the bovine rumen.

A common screen for plant antimicrobial compounds consists of separating plant extracts by paper or thin-layer chromatography (PC or TLC), exposing the ...

Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA)

Fire-derived, pyrogenic carbon (PyC), sometimes called black carbon (BC), is the carbonaceous solid residue of biomass and fossil fuel combustion, such as char and soot. PyC is ubiquitous in the environment due to its long persistence, and its abundance might even increase with the projected increase in global wildfire activity and the continued burning of fossil fuel. PyC is also increasingly produced from the industrial pyrolysis of organic wastes, which yields charred soil amendments (biochar). Moreover, the emergence of nanotechnology may also result in the release of PyC-like compounds to the environment. It is thus a high priority to reliably detect, characterize and quantify these charred materials in order to investigate their environmental properties and to understand their role in the carbon cycle.
Here, we present the benzene polycarboxylic acid (BPCA) method, which allows the simultaneous assessment of PyC&#39;s characteristics, quantity and isotopic composition (13C and 14C) on a molecular level. The method is applicable to a very wide range of environmental sample materials and detects PyC over a broad range of the combustion continuum, i.e., it is sensitive to slightly charred biomass as well as high temperature chars and soot. The BPCA protocol presented here is simple to employ, highly reproducible, as well as easily extendable and modifiable to specific requirements. It thus provides a versatile tool for the investigation of PyC in various disciplines, ranging from archeology and environmental forensics to biochar and carbon cycling research.

Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids BPCA

We present the benzene polycarboxylic acid (BPCA) method for assessing pyrogenic carbon (PyC) in the environment. The compound-specific approach uniquely provides simultaneous information about the characteristics, quantity and isotopic composition (13C and 14C) of PyC.

Fire-derived, pyrogenic carbon (PyC), sometimes called black carbon (BC), is the carbonaceous solid residue of biomass and fossil fuel combustion, such as ...

Preparation of Biopolymer Aerogels Using Green Solvents

Although the first reports on aerogels made by Kistler1 in the 1930s dealt with aerogels from both inorganic oxides (silica and others) and biopolymers (gelatin, agar, cellulose), only recently have biomasses been recognized as an abundant source of chemically diverse macromolecules for functional aerogel materials. Biopolymer aerogels (pectin, alginate, chitosan, cellulose, etc.) exhibit both specific inheritable functions of starting biopolymers and distinctive features of aerogels (80-99% porosity and specific surface up to 800 m2/g). This synergy of properties makes biopolymer aerogels promising candidates for a wide gamut of applications such as thermal insulation, tissue engineering and regenerative medicine, drug delivery systems, functional foods, catalysts, adsorbents and sensors. This work demonstrates the use of pressurized carbon dioxide (5 MPa) for the ionic cross linking of amidated pectin into hydrogels. Initially a biopolymer/salt dispersion is prepared in water. Under pressurized CO2 conditions, the pH of the biopolymer solution is lowered to 3 which releases the crosslinking cations from the salt to bind with the biopolymer yielding hydrogels. Solvent exchange to ethanol and further supercritical CO2 drying (10 - 12 MPa) yield aerogels. Obtained aerogels are ultra-porous with low density (as low as 0.02 g/cm3), high specific surface area (350 - 500 m2/g) and pore volume (3 - 7 cm3/g for pore sizes less than 150 nm).

A new way for the production of biopolymer-based aerogels by carbon dioxide (CO2) induced gelation is shown. The technique utilizes pressurized carbon dioxide (5 MPa) for the production of biopolymer hydrogels and supercritical CO2 (12 MPa) to convert gels into aerogels. The only solvents needed besides CO2 are water and ethanol.

Although the first reports on aerogels made by Kistler1 in the 1930s dealt with aerogels from both inorganic oxides (silica and others) and biopolymers ...

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While metal-based nanoparticles are associated with synthetic and environmental hassles, cellulose introduces a green, sustainable alternative for nanoparticle synthesis. Here, we present the chemical synthesis and separation procedures to produce new classes of hairy nanoparticles (bearing both amorphous and crystalline regions) and biopolymers based on wood fibers. Through periodate oxidation of soft wood pulp, the glucose ring of cellulose is opened at the C2-C3 bond to form 2,3-dialdehyde groups. Further heating of the partially oxidized fibers (e.g., T = 80 &#176;C) results in three products, namely fibrous oxidized cellulose, sterically stabilized nanocrystalline cellulose (SNCC), and dissolved dialdehyde modified cellulose (DAMC), which are well separated by intermittent centrifugation and co-solvent addition. The partially oxidized fibers (without heating) were used as a highly reactive intermediate to react with chlorite for converting almost all aldehyde to carboxyl groups. Co-solvent precipitation and centrifugation resulted in electrosterically stabilized nanocrystalline cellulose (ENCC) and dicarboxylated cellulose (DCC). The aldehyde content of SNCC and consequently surface charge of ENCC (carboxyl content) were precisely controlled by controlling the periodate oxidation reaction time, resulting in highly stable nanoparticles bearing more than 7 mmol functional groups per gram of nanoparticles (e.g., as compared to conventional NCC bearing &#60;&#60; 1 mmol functional group/g). Atomic force microscopy (AFM), transmission electron microscopy (TEM), and scanning electron microscopy (SEM) attested to the rod-like morphology. Conductometric titration, Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance (NMR), dynamic light scattering (DLS), electrokinetic-sonic-amplitude (ESA) and acoustic attenuation spectroscopy shed light on the superior properties of these nanomaterials.

Synthesis schemes to prepare highly stable wood fiber-based hairy nanoparticles and functional cellulose-based biopolymers have been detailed.

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While ...

A Simple, Low-cost, and Robust System to Measure the Volume of Hydrogen Evolved by Chemical Reactions with Aqueous Solutions

There is a growing research interest in the development of portable systems which can deliver hydrogen on-demand to proton exchange membrane (PEM) hydrogen fuel cells. Researchers seeking to develop such systems require a method of measuring the generated hydrogen. Herein, we describe a simple, low-cost, and robust method to measure the hydrogen generated from the reaction of solids with aqueous solutions. The reactions are conducted in a conventional one-necked round-bottomed flask placed in a temperature controlled water bath. The hydrogen generated from the reaction in the flask is channeled through tubing into a water-filled inverted measuring cylinder. The water displaced from the measuring cylinder by the incoming gas is diverted into a beaker on a balance. The balance is connected to a computer, and the change in the mass reading of the balance over time is recorded using data collection and spreadsheet software programs. The data can then be approximately corrected for water vapor using the method described herein, and parameters such as the total hydrogen yield, the hydrogen generation rate, and the induction period can also be deduced. The size of the measuring cylinder and the resolution of the balance can be changed to adapt the setup to different hydrogen volumes and flow rates.

The study of methods to generate on-demand hydrogen for fuel cells continues to grow in importance. However, systems to measure hydrogen evolution from the reaction of chemicals with water can be complicated and expensive. This article details a simple, low-cost, and robust method to measure the evolution of hydrogen gas.

There is a growing research interest in the development of portable systems which can deliver hydrogen on-demand to proton exchange membrane (PEM) ...

Preparation and Reactivity of a Triphosphenium Bromide Salt: A Convenient and Stable Source of Phosphorus(I)

We present herein the optimized synthesis of a triphosphenium bromide salt. Apart from being a versatile metathesis reagent, this unusually stable low-valent-phosphorus-containing compound acts as a useful P+ transfer agent. Unlike traditional methods employed to access low-coordinate phosphorus species which usually require pyrophoric phosphorus-containing precursors (white phosphorus, Tris(trimethylsilyl)phosphine, etc.), or harsh reducing agents (alkali metals, potassium graphite, etc.), the current approach does not involve pyrophoric or explosive reagents and can be done on large scales (&#62;20 g) in excellent yields by undergraduates with basic air-free synthetic training. The bromide counter ion is readily exchanged with other anions such as tetraphenyl borate (described herein) using typical salt metathesis reagents to obtain materials with desired properties and reactivities. The versatility of this P+ transfer approach is exemplified by the reactions of these triphosphenium precursors with an N-heterocyclic carbene and an anionic bisphosphine, each of which readily displace the neutral bisphosphine to give an NHC-stabilized phosphorus(I) cation and a phosphorus(I) containing zwitterion, respectively.

Preparation and Reactivity of a Triphosphenium Bromide Salt: A Convenient and Stable Source of PhosphorusI

The synthesis of a triphosphenium bromide salt is described and its use as a P+ transfer agent is outlined by reactions with an N-heterocyclic carbene and an anionic bisphosphine, yielding an NHC-stabilized P(I) cation and a P(I) containing zwitterion, respectively.

We present herein the optimized synthesis of a triphosphenium bromide salt. Apart from being a versatile metathesis reagent, this unusually stable ...

Synthesis of Core-shell Lanthanide-doped Upconversion Nanocrystals for Cellular Applications

Lanthanide-doped upconversion nanocrystals (UCNs) have attracted much attention in recent years based on their promising and controllable optical properties, which allow for the absorption of near-infrared (NIR) light and can subsequently convert it into multiplexed emissions that span over a broad range of regions from the UV to the visible to the NIR. This article presents detailed experimental procedures for high-temperature co-precipitation synthesis of core-shell UCNs that incorporate different lanthanide ions into nanocrystals for efficiently converting deep-tissue penetrable NIR excitation (808 nm) into a strong blue emission at 480 nm. By controlling the surface modification with biocompatible polymer (polyacrylic acid, PAA), the as-prepared UCNs acquires great solubility in buffer solutions. The hydrophilic nanocrystals are further functionalized with specific ligands (dibenzyl cyclooctyne, DBCO) for localization on the cell membrane. Upon NIR light (808 nm) irradiation, the upconverted blue emission can effectively activate the light-gated channel protein on the cell membrane and specifically regulate the cation (e.g., Ca2+) influx in the cytoplasm. This protocol provides a feasible methodology for the synthesis of core-shell lanthanide-doped UCNs and subsequent biocompatible surface modification for further cellular applications.

A protocol is presented for the synthesis of core-shell lanthanide-doped upconversion nanocrystals (UCNs) and their cellular applications for channel protein regulation upon near-infrared (NIR) light illumination.

Lanthanide-doped upconversion nanocrystals (UCNs) have attracted much attention in recent years based on their promising and controllable optical ...

Fabrication of Spherical and Worm-shaped Micellar Nanocrystals by Combining Electrospray, Self-assembly, and Solvent-based Structure Control

Micellar nanocrystals (micelles with encapsulated nanocrystals) have become an emerging major class of nanobiomaterials. We describe a method of fabricating micellar nanocrystals based on combining top-down electrospray, bottom-up self-assembly, and solvent-based structure control. This method involves first using electrospray to generate uniform ultrafine liquid droplets, each of which functions as a micro-reactor in which self-assembly reaction occurs forming micellar nanocrystals, with the structures (micelle shape and nanocrystal encapsulation) controlled by the organic solvent used. This method is largely continuous and produces high quality micellar nanocrystal products with an inexpensive structure control approach. By using a water-miscible organic solvent tetrahydrofuran (THF), worm-shaped micellar nanocrystals can be produced due to solvent-induced/facilitated micelle fusion. Compared with the common spherical micellar nanocrystals, worm-shaped micellar nanocrystals can offer minimized non-specific cellular uptake, thus enhancing biological targeting. By co-encapsulating multiple nanocrystals into each micelle, multifunctional or synergistic effects can be achieved. Current limitations of this fabrication method, which will be part of the future work, primarily include imperfect encapsulation in the micellar nanocrystal product and the incompletely continuous nature of the process.

The present work describes a method to fabricate micellar nanocrystals, an emerging major class of nanobiomaterials. This method combines top-down electrospray, bottom-up self-assembly, and solvent-based structure control. The fabrication method is largely continuous, can produce high quality products, and possesses an inexpensive means of structure control.