Name: determination of carbonyl functional groups in bio-oils by potentiometric titration: the faix method
Uploaded: 2017-02-07T14:00:00.000+00:00
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Description: Watch this Scientific Journal Video about Determination of Carbonyl Functional Groups in Bio-oils by Potentiometric Titration: The Faix Method at JoVE.com

This work was supported by the U.S. Department of Energy under Contract No. DE-AC36-08GO28308 with the National Renewable Energy Laboratory. Funding provided by U.S. DOE Office of Energy Efficiency and Renewable Energy Bioenergy Technologies Office. The U.S. Government retains and the publisher, by accepting the article for publication, acknowledges that the U.S. Government retains a nonexclusive, paid-up, irrevocable, worldwide license to publish or reproduce the published form of this work, or allow others to do so, for U.S. Government purposes.

Caution: Please review all relevant material safety data sheets (MSDS) before beginning. Ethanol is flammable. All applicable chemical handling procedures should be followed, as well as all applicable waste disposable and handling procedures.
1. Reagent Solutions
<ol>
	<li>Prepare the hydroxylamine hydrochloride solution (Solution A): Add 7.7 g of hydroxylamine hydrochloride and 50 mL of deionized water to a 250 mL volumetric flask. When all solids have dissolved, dilute to mark with ethanol. This results in a 0.55 M hydroxylamine hydrochloride solution in 80% (v/v) ethanol.</li>
	<li>Prepare the triethanolamine solution (Solution B): Add 17.4 mL of triethanolamine to a 250 mL volumetric flask. Add 10 mL of water, and then dilute up to the mark with ethanol. 95% ethanol can also be used if the addition of water is skipped. This results in a 0.48 M triethanolamine solution in 96 % (v/v) ethanol</li>
	<li>Prepare the hydrochloric acid solution. Either purchase 0.1 N solution or prepare using 10 mL concentrated HCl and 1 L water.</li>
</ol>
2. Bio-oil Sampling and Handling
<ol>
	<li>Make sure the oil sample is at room temperature prior to withdrawing a sample. Thoroughly homogenize (mix by shaking vigorously for at least 1 minute, and visually inspect the sample to ensure it is homogenous. Some bio-oils may require longer shaking times) bio-oil to obtain a representative sample.</li>
	<li>For the bio-oil sample, use 100 to 150 mg of bio-oil.</li>
	<li>Minimize exposure to oxygen and heat to prevent sample degradation prior to analysis.</li>
</ol>
3. Analytical Procedure
<ol>
	<li>Standardization of the Base Solution
	<ol>
		<li>Dry Na2CO3 primary standard in an oven at 105 &#176;C overnight to ensure a dry sample. Allow the Na2CO3 to cool to room temperature before weighing.</li>
		<li>Weigh 100 to 150 mg of sodium carbonate to a titration vessel, record the actual weight, add a stir bar and add enough water to cover the pH electrode bulb and junction.</li>
		<li>Transfer the sample to a titration vessel, washing the reaction vial several times separately with ethanol and water in proportions to make a final 80% solution of ethanol/water. Titrate with the acid solution using an automatic titrator to the endpoint. Record the endpoint. The endpoint is defined as the inflection point on the titration curve.</li>
		<li>Repeat the process twice to obtain three points.</li>
		<li>Use the average value as the normality of the acid solution.</li>
	</ol>
	</li>
	<li>Preparation of Titration Blanks
	<ol>
		<li>For Blank A: add 0.5 mL dimethyl sulfoxide (DMSO) to a 5 mL vial with a spin vane.</li>
		<li>Add 2 mL hydroxylamine hydrochloride solution (solution A).</li>
		<li>Add 2 mL triethanolamine solution (solution B).</li>
		<li>Cap tightly, place in preheated (80 &#176;C) heater block or water bath and stir for 2 hours.</li>
		<li>Titrate with acid solution using an automatic titrator to the endpoint, and record endpoint.</li>
		<li>For Blank B: If mineral acid is suspected to be present in sample, add 0.5 mL DMSO to 5 mL vial with a spin vane.</li>
		<li>Add 2 mL triethanolamine solution (solution B).</li>
		<li>Cap tightly and stir at 80 &#176;C for 2 hours.</li>
		<li>Transfer the sample to a titration vessel, washing the reaction vial several times separately with ethanol and water in proportions to make a final 80% solution of ethanol/water. Titrate with the acid solution using an automatic titrator to the endpoint. Record the endpoint.</li>
		<li>Repeat process three times to obtain three points.</li>
	</ol>
	</li>
	<li>Validation of the Method Using a Known Carbonyl
	<ol>
		<li>Weigh ~ 100 mg of 4-(benzyloxy)benzaldehyde (4-BBA) to a 5 mL vial, record the actual weight, add a spin vane.</li>
		<li>Add 0.5 mL DMSO.</li>
		<li>Dissolve the sample in 2 mL hydroxylamine hydrochloride solution (solution A).</li>
		<li>Add 2 mL triethanolamine solution (solution B).</li>
		<li>Close lid tightly and stir at 80 &#176;C for 2 hours.</li>
		<li>Transfer the sample to a titration vessel, washing the reaction vial several times separately with ethanol and water in proportions to make a final 80% solution of ethanol/water. Titrate with the acid solution using an automatic titrator to the endpoint. Record the endpoint.</li>
		<li>Repeat the process three times, to obtain three points.</li>
	</ol>
	</li>
	<li>Analysis of Bio-oil Using the Method
	<ol>
		<li>Weigh close to 100 mg of the bio-oil to 5 mL vial, record the actual weight and add a spin vane.</li>
		<li>Add 0.5 mL DMSO.</li>
		<li>Dissolve the sample in 2 mL hydroxylamine hydrochloride solution (solution A).</li>
		<li>Add 2 mL triethanolamine solution (solution B).</li>
		<li>Close lid tightly and stir at 80 &#176;C for 2 hours.</li>
		<li>Transfer the sample to a titration vessel, washing the reaction vial several times separately with ethanol and water in proportions to make a final 80% solution of ethanol/water. Titrate with the acid solution using an automatic titrator to the endpoint. Record the endpoint.</li>
		<li>Repeat the process three times, to obtain three points.</li>
		<li>Blank C: If mineral acid is suspected to be present in sample, weigh close to 100 mg of the bio-oil into a 5 mL vial, record the weight and add a spin vane.</li>
		<li>Add 0.5 mL DMSO.</li>
		<li>Add 2 mL triethanolamine solution (solution B).</li>
		<li>Cap tightly and stir at 80 &#176;C for 2 hours.</li>
		<li>Transfer the sample to a titration vessel, washing the reaction vial several times separately with ethanol and water in proportions to make a final 80% solution of ethanol/water. Titrate with the acid solution using an automatic titrator to the endpoint. Record the endpoint.</li>
		<li>Repeat process three times to obtain three points.</li>
	</ol>
	</li>
</ol>
4. Calculations
<ol>
	<li>Standardization of the Base Solution
	<ol>
		<li>Calculate the concentration of the acid solution (mol/L) using the following equation. The weight of the dry sodium carbonate in grams is w1, the purity is written as a fraction (i.e., 99% is 0.99), and the endpoint is in mL. 
		<img alt="Equation 1" src="/files/ftp_upload/55165/55165eq1.jpg" />&#160;&#160;&#160;</li>
	</ol>
	</li>
	<li>Validation of the Method Using a Known Carbonyl
	<ol>
		<li>Calculate the concentration of 4BBA (mol/L) in the sample using the following equation. The weight of the 4BBA in grams is w2, the 4BBA purity is written as a fraction (i.e., 99% is 0.99), the concentration of the acid solution (mol/L) is [acid], the triethanolamine/hydroxylamine&#8226;HCl blank endpoint is EPBA (the average value of three blanks, in mL), and the endpoint is EP (in mL). 
		<img alt="Equation 2" src="/files/ftp_upload/55165/55165eq2.jpg" />&#160;&#160;&#160;</li>
	</ol>
	</li>
	<li>Analysis of Bio-oil
	<ol>
		<li>Calculate the concentration of carbonyls in bio-oils [CO] (mmol/g-bio oil) using the following equation. The weight of the bio-oil in grams is w3, the concentration of the acid solution (mol/L) is [acid], the triethanolamine/hydroxylamine&#8226;HCl blank endpoint is EPBA (the average value of three blanks in mL), and the endpoint is EP (in mL). 
		<img alt="Equation 3" src="/files/ftp_upload/55165/55165eq3.jpg" />&#160;&#160;&#160;</li>
	</ol>
	</li>
	<li>Acid Correction 
	NOTE: The presence of mineral acids or organic acids with pKa &#60; 2 can cause artificially low carbonyl values due to the reaction of the acid with triethanolamine.
	<ol>
		<li>If this is suspected, perform the blanks detailed in sections 3.2.6 and 3.4.8. The weight of bio-oil in the sample in grams is w3, EP is the endpoint of the sample and EPBA is the triethanolamine/hydroxylamine blank. EPBB is the endpoint of Blank B, EPBC is endpoint of Blank C and the weight (g) of oil used in Blank C is wBC: 
		<img alt="Equation 4" src="/files/ftp_upload/55165/55165eq4.jpg" />&#160;&#160;&#160; 
		For bio-oil samples that contain mostly acetic acid, this is an unnecessary step.</li>
	</ol>
	</li>
</ol>

<ol>
	<li>Oasmaa, A., Kuoppala, E., Solantausta, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+pyrolysis+of+forestry+residue.+2.+physicochemical+composition+of+product+liquid.">Fast pyrolysis of forestry residue. 2. physicochemical composition of product liquid.</a> Energy Fuels. 17 (2), 433-443 (2003).</li><li>Olarte, M., 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=Stabilization+of+Softwood-Derived+Pyrolysis+Oils+for+Continuous+Bio-oil+Hydroprocessing.">Stabilization of Softwood-Derived Pyrolysis Oils for Continuous Bio-oil Hydroprocessing.</a> Top. Catal. 59 (1), 55-64 (2016).</li><li>Nicolaides, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The chemical characterization of pyrolytic oils. , (1984).</li><li>Oasmaa, A., Korhonen, J., Kuoppala, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+approach+for+stability+measurement+of+wood-based+fast+pyrolysis+bio-oils.">An approach for stability measurement of wood-based fast pyrolysis bio-oils.</a> Energy Fuels. 25 (7), 3307-3313 (2011).</li><li>Chen, C. L., Lin, S. Y., Dence, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Methods in Lignin Chemistry. , 446-457 (1992).</li><li>Scholze, B., Hanser, C., Meier, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+water-insoluble+fraction+from+fast+pyrolysis+liquids+(pyrolytic+lignin)+Part+II.+GPC,+carbonyl+groups,+and+13C-NMR.">Characterization of the water-insoluble fraction from fast pyrolysis liquids (pyrolytic lignin) Part II. GPC, carbonyl groups, and 13C-NMR.</a> J. Anal. Appl. Pyrolysis. 58-59, 387-400 (2001).</li><li>Bayerbach, R., Meier, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+water-insoluble+fraction+from+fast+pyrolysis+liquids+(pyrolytic+lignin).+Part+IV:+Structure+elucidation+of+oligomeric+molecules.">Characterization of the water-insoluble fraction from fast pyrolysis liquids (pyrolytic lignin). Part IV: Structure elucidation of oligomeric molecules.</a> J. Anal. Appl. Pyrolysis. 85 (1-2), 98-107 (2009).</li><li>Faix, O., Andersons, B., Zakis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+Carbonyl+Groups+of+Six+Round+Robin+Lignins.">Determination of Carbonyl Groups of Six Round Robin Lignins.</a> Holzforschung. 52, 268-272 (1998).</li><li>Black, S., Ferrell, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+Carbonyl+Groups+in+Pyrolysis+Bio-oils+Using+Potentiometric+Titration:+Review+and+Comparison+of+Methods.">Determination of Carbonyl Groups in Pyrolysis Bio-oils Using Potentiometric Titration: Review and Comparison of Methods.</a> Energy Fuels. 30 (2), 1071-1077 (2016).</li><li>Ferrell, J., 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=Standardization+of+Chemical+Analytical+Techniques+for+Pyrolysis+Bio-oil:+History,+Challenges,+and+Current+Status+of+Methods.">Standardization of Chemical Analytical Techniques for Pyrolysis Bio-oil: History, Challenges, and Current Status of Methods.</a> Biofuels, Bioprod. Biorefin. 10, 496-507 (2016).</li></ol>

The authors have nothing to disclose.

Representative titration curves are shown in Figure 2. A blank titration, as well as a titration for a pyrolysis oil sample, are shown. Furthermore, the first derivative of the titration curve (dpH/dV) is shown, which allows for easy recognition of the titration endpoint. The inset table on Figure 2 shows triplicate data for both pyrolysis oil and blank titrations, with average values and standard deviations. The endpoint values shown (in mL) are used in Section 4 to calculate the total ...

While pyrolysis bio-oils are comprised of a large variety of compounds and chemical functional groups, quantification of carbonyl groups is especially important. Carbonyls are known to be responsible for the instability of bio-oil during both storage1 and processing2. The titration method presented here is a simple technique which can reliably quantify the total carbonyl content of bio-oils. Only aldehyde and ketone functional groups are quantified using this method; carboxylic acid and lactone groups are not quantified.
For analysis of bio-oils, quantification of carbonyl groups by titration has traditionally been accomplished using the method of Nicolaides3. This method has been commonly used in the bio-oil literature4,5,6,7. This is a simple procedure where carbonyls are converted to the corresponding oxime (see Figure 1). The liberated HCl reacts with pyridine to force the equilibrium to completion. The conjugate acid of pyridine is titrated with a known amount of NaOH (base titrant). The number of equivalents of NaOH used is stoichiometrically equivalent to the moles of carbonyl present in the bio-oil.
The Nicolaides method, however, has several limitations. It can require reaction times in excess of 48 hours to reach completion. This severely limits sample throughput. It utilizes pyridine, which is toxic. Sample weights of 1 to 2 g are required. Sample weight used is dependent on the amount of hydroxylamine HCl present and the carbonyl content of the sample. If initial estimates of the sample weight used are incorrect, the titration has to be repeated.
Faix et al.8 developed a method that has been modified here to address the issues of the Nicolaides method. The reaction is carried out at 80 &#176;C for 2 hours, thereby increasing sample throughput. Pyridine has been replaced with triethanolamine, which is a less toxic chemical. The sample size can be reduced to 100 to 150 mg. The triethanolamine consumes the liberated HCl, driving the reaction to completion and the unconsumed triethanolamine is titrated directly. A secondary titration of the hydroxylamine is unnecessary. Comparison of these titration methods has shown that the Nicolaides method significantly underestimates carbonyl content of bio-oils9.
The method described here has been modified from the original method8 to be more applicable to the analysis of pyrolysis bio-oils. This method was developed for the analysis of raw pyrolysis bio-oils, but it has been successfully applied to other types of biomass-derived oils, including hydrotreated bio-oils. Additionally, this method has been used to monitor changes in carbonyl content during both aging and upgrading.

<table><tbody><tr><td>Analytical balance</td><td></td><td></td><td>accurate to 0.1 mg</td></tr><tr><td>dry block heater with magnetic stirrer, or hot water bath with magnetic stirrer</td><td></td><td></td><td></td></tr><tr><td>Automatic titrator</td><td></td><td></td><td>We used a Metrohm Titrando 809 automatic titrator, though other equivalent systems are acceptable</td></tr><tr><td>Deionized water</td><td></td><td></td><td></td></tr><tr><td>Ethanol (reagent grade)</td><td></td><td></td><td>CAS # 64-17-5</td></tr><tr><td>Hydroxylamine hydrochloride&amp;nbsp;</td><td></td><td></td><td>CAS # 5470-11-1</td></tr><tr><td>Triethanolamine&amp;nbsp;</td><td></td><td></td><td>CAS #102-71-6</td></tr><tr><td>Hydrochloric acid (37%)&amp;nbsp;</td><td></td><td></td><td>CAS # 7647-01-0</td></tr><tr><td>Sodium Carbonate (primary standard)&amp;nbsp;</td><td>SigmaAldrich</td><td>223484</td><td></td></tr><tr><td>4-(benzyloxy)benzaldehyde&amp;nbsp;</td><td></td><td></td><td>CAS # 4397-53-9</td></tr><tr><td>Dimethyl sulfoxide</td><td></td><td></td><td>CAS # 67-68-5</td></tr><tr><td>5 mL glass Reacti-vials with solid lid and teflon spinvane</td><td>Thermoscientific</td><td>TS-13223</td><td></td></tr><tr><td>200 mL volumetric flask</td><td></td><td></td><td></td></tr><tr><td>Volumetric or mechanical pipettes</td><td></td><td></td><td></td></tr></tbody></table>

determination of carbonyl functional groups in bio-oils by potentiometric titration: the faix method

Carbonyl compounds present in bio-oils are known to be responsible for bio-oil property changes upon storage and during upgrading. Specifically, carbonyls cause an increase in viscosity (often referred to as &#39;aging&#39;) during storage of bio-oils. As such, carbonyl content has previously been used as a method of tracking bio-oil aging and condensation reactions with less variability than viscosity measurements. Additionally, carbonyls are also responsible for coke formation in bio-oil upgrading processes. Given the importance of carbonyls in bio-oils, accurate analytical methods for their quantification are very important for the bio-oil community. Potentiometric titration methods based on carbonyl oximation have long been used for the determination of carbonyl content in pyrolysis bio-oils. Here, we present a modification of the traditional carbonyl oximation procedures that results in less reaction time, smaller sample size, higher precision, and more accurate carbonyl determinations. While traditional carbonyl oximation methods occur at room temperature, the Faix method presented here occurs at an elevated temperature of 80 &#176;C.

A typical titration curve consists of a single endpoint, as shown in Figure 2. Typical titrations for both a raw bio-oil sample, and a blank titration, are shown. As the endpoint lies at the inflection point in the titration curve; the endpoint can be easily identified by plotting the first derivative of the titration curve (shown on the right axis, dpH/dV, in Figure 2). Many automatic titration systems have software that calculates the derivative of the ...

Watch this Scientific Journal Video about Determination of Carbonyl Functional Groups in Bio-oils by Potentiometric Titration: The Faix Method at JoVE.com

Determination of Carbonyl Functional Groups in Bio-oils by Potentiometric Titration: The Faix Method

Here we present a potentiometric titration technique for accurately quantifying carbonyl compounds in pyrolysis bio-oils.

Carbonyl compounds present in bio-oils are known to be responsible for bio-oil property changes upon storage and during upgrading. Specifically, carbonyls ...

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Organic Chemistry

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Unit 1

Aldehydes and Ketones

Complexometric Titration, Precipitation Titration, and Gravimetry

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11.0K Views.  National Renewable Energy Laboratory. Here we present a potentiometric titration technique for accurately quantifying carbonyl compounds in pyrolysis bio-oils.

Video: Determination of Carbonyl Functional Groups in Bio-oils by Potentiometric Titration: The Faix Method

Identification of Fatty Acids in Bacillus cereus

The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the double bond. Changes in FA composition play a crucial role in the adaptation of bacteria to their environment. These modifications entail a change in the ratio of iso versus anteiso branched FAs, and in the proportion of unsaturated FAs relative to saturated FAs, with double bonds created at specific positions. Precise identification of the FA profile is necessary to understand the adaptation mechanisms of Bacillus species.
Many of the FAs from Bacillus are not commercially available. The strategy proposed herein identifies FAs by combining information on the retention time (by calculation of the equivalent chain length (ECL)) with the mass spectra of three types of FA derivatives: fatty acid methyl esters (FAMEs), 4,4-dimethyl oxazoline derivatives (DMOX), and 3-pyridylcarbinyl ester (picolinyl). This method can identify the FAs without the need to purify the unknown FAs.
Comparing chromatographic profiles of FAME prepared from Bacillus cereus with a commercial mixture of standards allows for the identification of straight-chain saturated FAs, the calculation of the ECL, and hypotheses on the identity of the other FAs. FAMEs of branched saturated FAs, iso or anteiso, display a constant negative shift in the ECL, compared to linear saturated FAs with the same number of carbons. FAMEs of unsaturated FAs can be detected by the mass of their molecular ions, and result in a positive shift in the ECL compared to the corresponding saturated FAs.
The branching position of FAs and the double bond position of unsaturated FAs can be identified by the electron ionization mass spectra of picolinyl and DMOX derivatives, respectively. This approach identifies all the unknown saturated branched FAs, unsaturated straight-chain FAs and unsaturated branched FAs from the B. cereus extract.

Identification of Fatty Acids in Bacillus cereus

We propose a protocol to identify fatty acids without the need to purify them. It combines information on the retention times with the mass spectra of three types of fatty acid derivatives: fatty acid methyl esters (FAMEs), 4,4-dimethyl oxazoline derivatives (DMOX), and 3-pyridylcarbinyl esters (picolinyl).

The Bacillus species contain branched chain and unsaturated fatty acids (FAs) with diverse positions of the methyl branch (iso or anteiso) and of the ...

Biochemistry

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The toolkit consists of multiple standard interchangeable parts (BioBricks)9 addressing the conversion of alkanes, regulation of gene expression and survival in toxic hydrocarbon-rich environments.
A three-step pathway for alkane degradation was implemented in E. coli to enable the conversion of medium- and long-chain alkanes to their respective alkanols, alkanals and ultimately alkanoic-acids. The latter were metabolized via the native &beta;-oxidation pathway. To facilitate the oxidation of medium-chain alkanes (C5-C13) and cycloalkanes (C5-C8), four genes (alkB2, rubA3, rubA4and rubB) of the alkane hydroxylase system from Gordonia sp. TF68,21 were transformed into E. coli. For the conversion of long-chain alkanes (C15-C36), theladA gene from Geobacillus thermodenitrificans was implemented. For the required further steps of the degradation process, ADH and ALDH (originating from G. thermodenitrificans) were introduced10,11. The activity was measured by resting cell assays. For each oxidative step, enzyme activity was observed.
To optimize the process efficiency, the expression was only induced under low glucose conditions: a substrate-regulated promoter, pCaiF, was used. pCaiF is present in E. coli K12 and regulates the expression of the genes involved in the degradation of non-glucose carbon sources.
The last part of the toolkit - targeting survival - was implemented using solvent tolerance genes, PhPFD&alpha; and &beta;, both from Pyrococcus horikoshii OT3. Organic solvents can induce cell stress and decreased survivability by negatively affecting protein folding. As chaperones, PhPFD&alpha; and &beta; improve the protein folding process e.g. under the presence of alkanes. The expression of these genes led to an improved hydrocarbon tolerance shown by an increased growth rate (up to 50%) in the presences of 10% n-hexane in the culture medium were observed.
Summarizing, the results indicate that the toolkit enables E. coli to convert and tolerate hydrocarbons in aqueous environments. As such, it represents an initial step towards a sustainable solution for oil-remediation using a synthetic biology approach.

A sustainable auto regulating bacterial system for the remediation of oil pollutions was designed using standard interchangeable DNA parts (BioBricks). An engineered E. coli strain was used to degrade alkanes via &beta;-oxidation in toxic aqueous environments. The respective enzymes from different species showed alkane degradation activity. Additionally, an increased tolerance to n-hexane was achieved by introducing genes from alkane-tolerant bacteria.

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The ...

Bioengineering

Analyzing the Photo-oxidation of 2-propanol at Indoor Air Level Concentrations Using Field Asymmetric Ion Mobility Spectrometry

We demonstrate a versatile protocol to be used for determining the effectiveness of photocatalysts in degrading indoor air concentration (ppb) volatile organic carbons (VOCs), illustrating this with a titanium dioxide based catalyst, and the VOC 2-propanol. The protocol takes advantage of field asymmetric ion mobility spectroscopy (FAIMS), an analysis tool that is capable of continuously identifying and monitoring the concentration of VOCs such as 2-propanol and acetone at the ppb level. The continuous nature of FAIMS allows detailed kinetic analysis, and long-term reactions, offering a significant advantage over gas chromatography, a batch process traditionally used in air purification characterization. The use of FAIMS in photocatalytic air purification has only recently been used for the first time, and with the protocol illustrated here, the flexibility in allowing alternative VOCs and photocatalysts to be tested using comparable protocols offers a unique system to elucidate photocatalytic air purification reactions at low concentrations.

A protocol for determining the effectiveness of photocatalysts in degrading indoor air concentration (ppb) model volatile organic carbons such as 2-propanol is described.

We demonstrate a versatile protocol to be used for determining the effectiveness of photocatalysts in degrading indoor air concentration (ppb) volatile ...

Original Experimental Approach for Assessing Transport Fuel Stability

The study of fuel oxidation stability is an important issue for the development of future fuels. Diesel and kerosene fuel systems have undergone several technological changes to fulfill environmental and economic requirements. These developments have resulted in increasingly severe operating conditions whose suitability for conventional and alternative fuels needs to be addressed. For example, fatty acid methyl esters (FAMEs) introduced as biodiesel are more prone to oxidation and may lead to deposit formation. Although several methods exist to evaluate fuel stability (induction period, peroxides, acids, and insolubles), no technique allows one to monitor the real-time oxidation mechanism and to measure the formation of oxidation intermediates that may lead to deposit formation. In this article, we developed an advanced oxidation procedure (AOP) based on two existing reactors. This procedure allows the simulation of different oxidation conditions and the monitoring of the oxidation progress by the means of macroscopic parameters, such as total acid number (TAN) and advanced analytical methods like gas chromatography coupled to mass spectrometry (GC-MS) and Fourier Transform Infrared - Attenuated Total Reflection (FTIR-ATR). We successfully applied AOP to gain an in-depth understanding of the oxidation kinetics of a model molecule (methyl oleate) and commercial diesel and biodiesel fuels. These developments represent a key strategy for fuel quality monitoring during logistics and on-board utilization.

Oxidation stability of transport fuels has become a concern for future fuel development. This work presents an original methodology developed by IFP Energies Nouvelles for assessing fuel stability using two different reactors. This methodology was successfully applied to gain an in-depth understanding of the oxidation kinetics and pathways of model molecules and commercial fuels.

The study of fuel oxidation stability is an important issue for the development of future fuels. Diesel and kerosene fuel systems have undergone several ...

Using Capillary Electrophoresis to Quantify Organic Acids from Plant Tissue: A Test Case Examining Coffea arabica Seeds

Carboxylic acids are organic acids containing one or more terminal carboxyl (COOH) functional groups. Short chain carboxylic acids (SCCAs; carboxylic acids containing three to six carbons), such as malate and citrate, are critical to the proper functioning of many biological systems, where they function in cellular respiration and can serve as indicators of cell health. In foods, organic acid content can have significant impact on taste, with increased SCCA levels resulting in a sour or &#34;acid&#34; taste. Because of this, methods for the rapid analysis of organic acid levels are of particular interest to the food and beverage industries. Unfortunately, however, most methods used for SCCA quantification are dependent on time-consuming protocols requiring the derivatization of samples with hazardous reagents, followed by costly chromatographic and/or mass spectrometric analyses. This method details an alternate method for the detection and quantification of organic acids from plant material and food samples using free zonal capillary electrophoresis (CZE), sometimes simply referred to as capillary electrophoresis (CE). CZE provides a cost-effective method for measuring SCCAs with a low limit of detection (0.005 mg/ml). This article details the extraction and quantification of SCCAs from plant samples. While the method provided focuses on measurement of SCCAs from coffee beans, the method provided can be applied to multiple plant-based food materials.

Using Capillary Electrophoresis to Quantify Organic Acids from Plant Tissue: A Test Case Examining Coffea arabica Seeds

This article presents a method for the detection and quantification of organic acids from plant material using free zonal capillary electrophoresis. An example of the potential application of this method, determining the effects of a secondary fermentation on organic acid levels in coffee seeds, is provided.

Carboxylic acids are organic acids containing one or more terminal carboxyl (COOH) functional groups. Short chain carboxylic acids (SCCAs; carboxylic ...

Extraction of Lignin with High &#946;-O-4 Content by Mild Ethanol Extraction and Its Effect on the Depolymerization Yield

Lignin valorization strategies are a key factor for achieving more economically competitive biorefineries based on lignocellulosic biomass. Most of the emerging elegant procedures to obtain specific aromatic products rely on the lignin substrate having a high content of the readily cleavable &#946;-O-4 linkage as present in the native lignin structure. This provides a miss-match with typical technical lignins that are highly degraded and therefore are low in &#946;-O-4 linkages. Therefore, the extraction yields, and the quality of the obtained lignin are of utmost importance to access new lignin valorization pathways. In this manuscript, a simple protocol is presented to obtain lignins with high &#946;-O-4 content by relatively mild ethanol extraction that can be applied to different lignocellulose sources. Furthermore, analysis procedures to determine the quality of the lignins are presented together with a depolymerization protocol that yields specific phenolic 2-arylmethyl-1,3-dioxolanes, which can be used to evaluate the obtained lignins. The presented results demonstrate the link between lignin quality and potential for the lignins to be depolymerized into specific monomeric aromatic chemicals. Overall, the extraction and depolymerization demonstrates a trade-off between the lignin extraction yield and the retention of the native aryl-ether structure and thus the potential of the lignin to be used as the substrate for the production of chemicals for higher-value applications.

Here, we present a protocol to perform ethanol extraction of lignin from several biomass sources. The effect of the extraction conditions on the lignin yield and &#946;-O-4 content are presented. Selective depolymerization is performed on the obtained lignins to obtain high aromatic monomer products.

Lignin valorization strategies are a key factor for achieving more economically competitive biorefineries based on lignocellulosic biomass. Most of the ...

Characterizing Lewis Pairs Using Titration Coupled with In Situ Infrared Spectroscopy

Lewis acid-activation of carbonyl-containing substrates is a fundamental basis for facilitating transformations in organic chemistry. Historically, characterization of these interactions has been limited to models equivalent to stoichiometric reactions. Here, we report a method utilizing in situ infrared spectroscopy to probe the solution interactions between Lewis acids and carbonyls under synthetically relevant conditions. Using this method, we were able to identify 1:1 complexation between GaCl3 and acetone and a highly ligated complex for FeCl3 and acetone. The impact of this technique on mechanistic understanding is illustrated by application to the mechanism of Lewis acid-mediated carbonyl-olefin metathesis in which we were able to observe competitive binding interactions between substrate carbonyl and product carbonyl with the catalyst.

Here, we present a method for the observation of solution interactions between Lewis acids and bases by employing in situ infrared spectroscopy as a detector for titration under synthetically relevant conditions. By examining solution interactions, this method represents a complement to X-ray crystallography, and provides an alternative to NMR spectroscopy.

Lewis acid-activation of carbonyl-containing substrates is a fundamental basis for facilitating transformations in organic chemistry. Historically, ...

Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

Fast pyrolysis is being increasingly applied in commercial plants worldwide. They run exclusively on woody biomass, which has favorable properties for conversion with fast pyrolysis. In order to increase the synergies of food production and the energetic and/or material use of biomass, it is desirable to utilize residues from agricultural production, e.g., straw. The presented method is suitable for converting such a material on an industrial scale. The main features are presented and an example of mass balances from the conversion of several biomass residues is given. After conversion, fractionated condensation is applied in order to retrieve two condensates &#8212; an organic-rich and an aqueous-rich one. This design prevents the production of fast pyrolysis bio-oil that exhibits phase separation. A two phase bio-oil is to be expected because of the typically high ash content of straw biomass, which promotes the production of water of reaction during conversion.
Both fractionated condensation and the use of biomass with high ash content demand a careful approach for establishing balances. Not all kind of balances are both meaningful and comparable to other results from the literature. Different balancing methods are presented, and the information that can be derived from them is discussed.

A procedure for thermochemical conversion of biomass residues is presented that aims at maximizing the yield of liquid products (fast pyrolysis). It is based on a technology proven on an industrial scale and especially suitable for treating a straw type of biomass.

Fast pyrolysis is being increasingly applied in commercial plants worldwide. They run exclusively on woody biomass, which has favorable properties for ...

Fractionation of Lignocellulosic Biomass using the OrganoCat Process

The shift from a petroleum-based to a more sustainable and bio-based economy requires the development of new refinery concepts to maintain the supply of raw materials and energy. For these novel and sustainable biorefinery concepts, it is important to use catalysts and solvents that are aligned with the principles of Green Chemistry. Therefore, the implementation of biogenic alternatives can be a promising solution. The lignocellulose pretreatment and fractionation process presented herein-OrganoCat-is an integrated fractionation of lignocellulose into its main components using biogenic acids such as 2,5-furandicarboxylic acid as catalyst. Hemicelluloses and other non-cellulosic polysaccharides are selectively depolymerized by the diluted acid and dissolved, while the crystalline cellulose remains in the solid pulp. In the presence of a second organic phase consisting of biogenic 2-methyltetrahydrofuran, disentangled lignin is extracted in situ. The process allows for the efficient fractionation of the three main components-lignin, cellulose, and non-cellulosic sugars. This helps to focus on the quality of the lignin, the improvement of enzymatic hydrolysis of the cellulose-enriched pulp, and the mild non-cellulosic sugar extraction with low degradation.

OrganoCat is a method for the pretreatment and fractionation of lignocellulose under mild conditions into lignin, fermentable sugars, and cellulose pulp. In a biogenic, biphasic solvent system of water and 2-methyltetrahydrofuran with 2,5-furancarboxylic acid as catalyst, the OrganoCat products are separated in situ for straightforward product recovery.

The shift from a petroleum-based to a more sustainable and bio-based economy requires the development of new refinery concepts to maintain the supply of ...

Ultrafast Lignin Extraction from Unusual Mediterranean Lignocellulosic Residues

Pretreatment is still the most expensive step in lignocellulosic biorefinery processes. It must be made cost-effective by minimizing chemical requirements as well as power and heat consumption and by using environment-friendly solvents. Deep eutectic solvents (DESs) are key, green, and low-cost solvents in sustainable biorefineries. They are transparent mixtures characterized by low freezing points resulting from at least one hydrogen bond donor and one hydrogen bond acceptor. Although DESs are promising solvents, it is necessary to combine them with an economic heating technology, such as microwave irradiation, for competitive profitability. Microwave irradiation is a promising strategy to shorten the heating time and boost fractionation because it can rapidly attain the appropriate temperature. The aim of this study was to develop a one-step, rapid method for biomass fractionation and lignin extraction using a low-cost and biodegradable solvent. 
In this study, a microwave-assisted DES pretreatment was conducted for 60 s at 800 W, using three kinds of DESs. The DES mixtures were facilely prepared from choline chloride (ChCl) and three hydrogen-bond donors (HBDs): a monocarboxylic acid (lactic acid), a dicarboxylic acid (oxalic acid), and urea. This pretreatment was used for biomass fractionation and lignin recovery from marine residues (Posidonia leaves and aegagropile), agri-food byproducts (almond shells and olive pomace), forest residues (pinecones), and perennial lignocellulosic grasses (Stipa tenacissima). Further analyses were conducted to determine the yield, purity, and molecular weight distribution of the recovered lignin. In addition, the effect of DESs on the chemical functional groups in the extracted lignin was determined by Fourier-transform infrared (FTIR) spectroscopy. The results indicate that the ChCl-oxalic acid mixture affords the highest lignin purity and the lowest yield. The present study demonstrates that the DES-microwave process is an ultrafast, efficient, and cost-competitive technology for lignocellulosic biomass fractionation.

Deep eutectic solvent-based, microwave-assisted pretreatment is a green, fast, and efficient process for lignocellulosic fractionation and high-purity lignin recovery.

Pretreatment is still the most expensive step in lignocellulosic biorefinery processes. It must be made cost-effective by minimizing chemical requirements ...

Determination of Carbonyl Functional Groups in Bio-oils by Potentiometric Titration: The Faix Method

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Using Capillary Electrophoresis to Quantify Organic Acids from Plant Tissue: A Test Case Examining Coffea arabica Seeds

Extraction of Lignin with High β-O-4 Content by Mild Ethanol Extraction and Its Effect on the Depolymerization Yield

Characterizing Lewis Pairs Using Titration Coupled with In Situ Infrared Spectroscopy

Fast Pyrolysis of Biomass Residues in a Twin-screw Mixing Reactor

Fractionation of Lignocellulosic Biomass using the OrganoCat Process

Ultrafast Lignin Extraction from Unusual Mediterranean Lignocellulosic Residues