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Paper mulberry, a fast-growing and vigorous plant, is a potential substrate for producing lignocellulosic bioethanol and an important renewable alternative to fossil fuels. In order to improve the economic feasibility of ethanol production from paper mulberry, H3PO4/H2O2 pretreatment was selected as the most suitable pretreatment method that could produce the highest glucose concentration (131 g/L) compared with other pretreatments (73.2–89.3 g/L) at high solid loading. Whereafter, the final solid loading of H3PO4/H2O2 pretreatment was significantly increased to 40% (w/v) without any decrease in the final glucose concentration. Finally, a novel Fed-nonisothermal-simultaneous saccharification and fermentation was constructed using H3PO4/H2O2 pretreated paper mulberry, which bypassed the inhibition caused by paper mulberry solid and high temperature on the traditional simultaneous saccharification and fermentation, improving ethanol concentration (63.9 g/L), ethanol productivity (1.33 g/L/h) and ethanol yield (0.160 g/g-biomass) by 30.4%, 30.4% and 30.1%, respectively, compared to those obtained from SSF (simultaneous saccharification and fermentation) process. Thus, we have opened up a novel way to produce ethanol or other biofuels using the paper mulberry as an outstanding alternative substrate.

This work presents a novel development that exploits the concept of in-situ gas-separation together with a specific enzymatic colorimetric detection to produce a portable biosensor called “Blood Alcohol Micro-pad” for direct quantitation of ethanol in whole blood. The thin square device (25 mm × 25 mm × 1.8 mm) comprises two layers of patterned filter paper held together with a double-sided mounting tape with an 8-mm circular hole (the headspace). In operation, the reagent is deposited on one layer and covered with sticky tape. Then 8 μL of a blood sample is dispensed onto the opposite layer and covered with sticky tape. Diffusion of ethanol across the 1.6 mm narrow headspace permits selective detection of ethanol by the enzymatic reagents deposited on the opposite layer. This reagent zone contains alcohol oxidase, horseradish peroxidase and 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, as the chromogenic reagent. The color intensity, measured from the recorded digital image, resulting from the enzymatic assay of ethanol, correlates with the concentration of blood alcohol. The results obtained with spiked mice and sheep blood samples, using an external calibration in the range of 1–120 mg dL−1ethanol, gave recoveries of 93.2–104.4% (n = 12). The “Blood Alcohol Micro-pad” gave good precision with %RSD

Thermophilic methane fermentation was a valid approach for handling the stillage eluted from ethanol fermentation of waste paper and kitchen waste. The wide organic loading rate (OLR) range (2-14 g VTS/(L d)) for stable performance and relatively high energy recovery efficiency (79.0%) were achieved, and OLR of 8 g VTS/(L d) was optimum for achievement of highest biogas evolution and VTS removal efficiency. Microbial community analysis revealed that hydrolysis of cellulose was the critical step for methane production from the stillage.

A novel visual and fluorometric-sensor paper is prepared for the monitoring of atmospheric humidity and ethanol. The paper contains a combination of copper nanoclusters (CuNCs) and carbon dots (CDs), immobilized on a filter paper. This makes it a dual-emission system. The visual sensor shows bright fluorescence and has a diameter of ∼8 cm with uniform and homogeneous surface. The detection mechanism of the sensing paper is based on the fluorescence intensity variation of CuNCs in different environments. Desirable linear relationship is observed between fluorescence intensity ratio (red/blue) and humidity. Visible color transformation of this sensor paper is observed in the humidity range ∼40-80%, implying utility for easy and facile atmospheric humidity determination. Furthermore, due to the water and ethanol have different impacts on the spectrum, this paper-based fluorescent probe shows excellent response to ethanol. Therefore dual-emitting sensing paper, when suitably calibrated, shows promise for visual and fluorometric sensing platforms aimed at humidity or ethanol detection.

Paper is a perfect example of the circular economy as it remains the furthermost recycled product in Europe, creating significant environmental benefits and raw materials resources to the industry. Indeed, maintaining a consistent level of quality whilst limiting the environmental footprint of the product has become a major challenge for the industry. In this direction, paper is proving to be the promising feedstock for biorefinery and biomaterials. The future of paper recycling is slowly going beyond fibre recovery to address the needs of other industries because for the earth's environmental well-being various paper products need to be recycled and reused persistently. In this article, we outline the ambitious use of wastepaper (WP) for high-value applications such as; production of cellulose nanocrystals (CNC), composite reinforcement, high performance electrical components and biofuels.

This study investigated the extent of enzyme savings that could be achieved by recycling enzyme-containing supernatant or whole fermentation broth from simultaneous saccharification and fermentation (SSF) of paper waste sludge (PS). Kinetic models were developed to identify the preferred enzyme recycling method. Kinetic models accounted for the loss of enzyme activity with time and the effect of enzyme supplementation on ethanol concentrations and predicted the performance of PS fermentation with acceptable accuracy. Process performance of SSF cultures was maintained during successive recycling steps by recycling either the enzyme-containing supernatant or whole broth without resorting to enzyme separation methods. Recycling of enzymes in the clarified supernatant during 5 L bioreactor fermentations allowed up to 23% savings on the overall enzyme loading. Recycling enzymes in the supernatant further increased the ethanol concentration in the fermentation broth from 40 to 70 g/L. The combined advantages of high ethanol concentrations and savings on enzyme consumption, opens up the possibility of achieving significant improvements in the economic viability of ethanol production from lignocellulosic substrates.

Efficient ethanol production from waste paper requires the addition of expensive nutrients. To reduce the production cost of ethanol from waste paper, a study on how to produce ethanol efficiently by adding kitchen waste (potentially as a carbon source, nutrient source, and acidity regulator) to waste paper was performed and a process of successive liquefaction, presaccharification, and simultaneous saccharification and fermentation (L+PSSF) was developed. The individual saccharification performances of waste paper and kitchen waste were not influenced by their mixture. Liquefaction of kitchen waste at 90°C prior to presaccharification and simultaneous saccharification and fermentation (PSSF) was essential for efficient ethanol fermentation. Ethanol at concentrations of 46.6 or 43.6g/l was obtained at the laboratory scale after fermentation for 96h, even without pH adjustment and/or the addition of extra nutrients. Similarly, ethanol at a concentration of 45.5g/l was obtained at the pilot scale after fermentation for 48h. The ethanol concentration of L+PSSF of the mixture of waste paper and kitchen waste was comparable to that of PSSF of waste paper with added nutrients (yeast extract and peptone) and pH adjustment using H2SO4, indicating that kitchen waste is not only a carbon source but also an excellent nutrient source and acidity regulator for fermentation of the mixture of waste paper and kitchen waste.

As traditional barriers are vulnerable to organic solvents or surfactants, a new fluoroalkyl-silica barrier with superior confining ability was developed for paper-based analytical devices (PADs). The as-prepared fluoroalkyl-silica barrier is able to contain surfactant solutions and pure organic solvents even alcohols. Based on the fluoroalkyl-silica barrier, PADs associated with condensing-enrichment effect were explored for Ni(II) assays involving ethanol and bacteria assays including sodium dodecyl sulfate (SDS) and chloroform. The exhibited work has the potential to significantly expand the applications of paper-based microfluidics or assays to include process/analysis involving organic solvents or surfactant solutions.

In this study we investigated the technical feasibility of convert paper mill sludge into fuel ethanol. This involved the removal of mineral fillers by using either chemical pretreatment or mechanical fractionation to determine their effects on cellulose hydrolysis and fermentation to ethanol. In addition, we studied the effect of cationic polyelectrolyte (as accelerant) addition and hydrogen peroxide pretreatment on enzymatic hydrolysis and fermentation. We present results showing that removing the fillers content (ash and calcium carbonate) from the paper mill sludge increases the enzymatic hydrolysis performance dramatically with higher cellulose conversion at faster rates. The addition of accelerant and hydrogen peroxide pretreatment further improved the hydrolysis yields by 16% and 25% (g glucose / g cellulose), respectively with the de-ashed sludge. The fermentation process of produced sugars achieved up to 95% of the maximum theoretical ethanol yield and higher ethanol productivities within 9h of fermentation.

Herein, to apply porous gold electrode for the electrooxidation of ethanol, we proposed a simple and facile method for the synthesis of paper-based porous gold (PAu) with large surface area using gold nanoparticles (AuNPs) and filter paper. The results showed that Pd/PAu exhibited a large electrochemical surface because of pore size of 100 nm and high electrochemical reaction compared to smooth Au foil and PAu. The as-prepared Pd/PAu electrode exhibited high activity and stability for ethanol electrooxidation in alkaline solution and was reusable even after 100 cycles.

This study optimized the simultaneous saccharification and citric acid (CA) production from banana pseudostem (BP). Thereafter, kinetic assessment of Aspergillus brasiliensis growth and CA production were determined for the optimum conditions using fresh water (SSF

We developed a convenient method by coupling frontal elution paper chromatography with desorption corona beam ionization mass spectrometry (DCBI-MS) for rapid determination of chlorphenamine added in herbal medicines or dietary supplements. In this method, the ethanol extract of the herbal products was spotted directly onto an isosceles triangular filter paper sheet, and then the paper sheet was developed under strong elution condition with the sample zone migrating at the solvent front. The analyte was finally condensed at the V-shaped tip which could then be placed under the visible plasma beam of DCBI for ionization. The overall procedure took less than 5 min. The frontal elution paper chromatography on a triangular plate used in this work improved the signal intensity of chlorphenamine by 30-fold due to the analyte condensing at the tip and the reduction of the background suppression. Furthermore, the paper sheet also functioned as a filter in the analysis of solid or powder samples, which can increase the analytical throughput by omitting the step of centrifugation. The proposed method in current study was successfully applied in the determination of chlorphenamine in herbal medicines. Chlorphenamine was detected in four of the twelve types of herbal medicines examined in this study. The limit of detection was 200 ng/mL (2.0 ng absolute) in full-scan positive-ion mode and the linear range was from 5.0 μg/mL to 50 μg/mL with satisfactory linear coefficient (R(2) (the square of the correlation coefficient)=0.895). Good reproducibility was achieved with relative standard deviations (RSDs) less than 15.0% and the recoveries of chlorphenamine ranged from 84.3 to 90.6%.

Pulping and papermaking of Leucaena diversifolia by soda-anthraquinone-ethanol was studied using an experimental design in order to investigate the effects of cooking variables: temperature, time, soda concentration, ethanol concentration and wash-disintegrate temperature on the pulp yield and the physico-chemical characteristics of paper sheets (tensile index, burst index, tear index and brightness). Previously, in order to assess the potential of plants of this raw material grown over short periods, its results were compared with those of other leucaena varieties and the best crop among three grown for 1–3 years was selected. The results were evaluated using the response surface methodology with a view to identifying the most suitable operating conditions. In accordance with biomass production and the features of the raw materials and cellulose pulp obtained, the L. diversifolia grown for 2 years was found to be the most suitable choice for obtaining pulp and paper among the five leucaena varieties examined. Suitable physical characteristics of paper sheets (tensile index, burst and tear index) and acceptable yield pulping and brightness could be obtained by operating at medium temperature, active alkali concentration, pulping time, ethanol concentration and wash-disintegrate temperature.

This work examined the simultaneous saccharification and fermentation (SSF) process for the biological conversion of pulp & paper sludge (PPS) and monosodium glutamate waste liquor (MGWL) into ethanol using Saccharomyces cerevisiae CICC 1001 in batch condition. At the SSF conditions of sterilization, pH 6.0, 6% TS (total solid), and adding nutrient solution, the highest ethanol yield of 42.5 g L−1 was obtained at the enzyme loading of 40 A.U. (activity unit) g−1 VSfed (volatile solid); only if the pH value was changed to 4.5, the ethanol yield reduced by 47%; only if the TS concentration was changed to 3%, the ethanol yield decreased by 11%; only if the substrates were not sterilized, 5% decrease occurred to the ethanol production; only if no nutrient solution was added to the reactor, 97% reduction of ethanol yield happened; the extent of influence of the impact factors ranked in the order of nutrient solution > pH > TS concentration > sterilization. The Fourier Transform Infrared (FTIR) spectra of substrates after SSF further verified the results of process performances. Therefore, this SSF of PPS and MGWL reduced the residue streams and land-filling costs while benefiting in ethanol production.

The hydrolytic kinetics of mixtures of cotton gin waste (CGW) and recycled paper sludge (RPS) at various initial enzyme concentrations of Spezyme™ AO3117 was investigated. The experiments showed that the concentrations of reducing sugars and the conversions of the mixtures increased with increasing initial enzyme concentration. The reducing sugar concentration and conversion of the mixture of 75% CGW and 25% RPS were higher than those of the mixture of 80% CGW and 20% RPS. The conversion of the former can reach 73.8% after a 72-h hydrolysis at the initial enzyme loading of 17.4 Filter Paper Unit (FPU)/g substrate. A three-parameter kinetic model based on enzyme deactivation and its analytical expression were derived. Using nonlinear regression, the parameters of the model were determined for the experimental data of hydrolytic kinetics of the mixtures. Based on this kinetic model of hydrolysis, two profit rate models, representing two kinds of operating modes with and without feedstock recycling, were developed. Using the profit rate models, the optimal enzyme loading and hydrolytic time can be predicted for the maximum profit rate in ethanol production according to the costs of enzyme and operation, enzyme loading, and ethanol market price. Simulated results from the models based on the experimental data of hydrolysis of the mixture of 75% CGW and 25% RPS showed that use of a high substrate concentration and an operating mode with feedstock recycle can greatly increase the profit rate in ethanol production. The results also demonstrated that the hydrolysis at a low enzyme loading is economically required for systematic optimization of ethanol production.

High-solid paper sludge was investigated for its potential to produce renewable biofuel. As a pretreatment method without consuming a large amount of energy, mechanical grinding for a comparatively short time followed by chemical swelling was used for optimal enzyme saccharification and ethanol production from paper sludge. Chemical swelling by phosphoric acid was more favorable than ball mill grinding. However, sequential pretreatment system using ball milling for 2 min and then phosphoric acid swelling for 1 h enhanced the enzyme saccharification rate and the reducing sugar productivity, 84.1% and 28.1 mg/g/h, respectively. The simultaneous saccharification and fermentation (SSF) from the sequential pretreatment system resulted to 81.5% ethanol conversion rate, the productivity being 1.27 g/L/h compared to untreated raw paper sludge which gave 54.3% ethanol conversion rate and 0.424 g/L/h productivity. Our work shows that consecutive mechanical grinding and chemical swelling are effective pretreatment methods for the enhancement of enzyme saccharification and efficient ethanol production from paper sludge.

During past decades, the considerable efforts have been made to utilize lignocellulose as biomass feedstock for the optimal production of bio-ethanol as an alternative source of fuel. In this work, the effective bioconversion of paper sludge to ethanol was investigated by using Zymomonas mobilis NBRC 13756. Major components of the raw paper sludge were carbohydrates and ash. Fermentation utilizing strains of Z. mobilis instead of traditional yeasts and the use of simultaneous saccharification and fermentation (SSF) process have been proposed due to their high ethanol yields with cost-effectiveness. Free cells of Z. mobilis resulted in no ethanol production even after 24 h of incubation because cells’ growth was inhibited by metal ions contained in the paper sludge. The application of SSF with Ca-alginate-immobilized cells of Z. mobilis was performed and 18 g/L of ethanol was obtained after 48 h of incubation at an initial paper sludge concentration of 200 g/L. In addition, the repeated batch fermentation of Ca-alginate-immobilized Z. mobilis cells was attempted for producing ethanol up to run 4. Our work suggests that Ca-alginate-immobilized cells of Z. mobilis could effectively produce ethanol from paper sludge not only under the batch fermentation but also under the repeated batch fermentation.

A central composite factorial design was used to examine the influence of independent variables of the ethanol pulping of wheat straw (viz. processing temperature and time, and ethanol concentration) on the yield and Shopper–Riegler index of the resulting pulp, and on various physical properties of paper sheets (viz. breaking length, stretch, burst index, tear index and brightness) obtained from it. By using the BMDP software suite, equations that relate each dependent variable to the different independent variables were obtained, and these reproduce the experimental results for yield, Shopper–Riegler index and brightness with errors less than 10%, and all other properties of the paper sheets to within 25%, over the temperature (140–180 °C), time (60–120 min) and ethanol concentration (40–80%) ranges allowing the production of pulps spanning a broad range of yields (from 37–79%). Ensuring the optimal breaking length (3550 m), stretch (1.42%), burst index (1.62 kN/g) and tear index (4.77 mNm 2 /g) entails using high temperature and medium-to-low ethanol concentration and time values; these conditions decrease the yield and brightness below their optimum levels (78.91 and 30.50%, respectively). However, high temperature, in combination with low ethanol concentration and short time, can be used to save solvent in the operations of recovery and recycling and to increase production. The values thus obtained for the dependent variables differ by less than 22% from their predicted optimum levels in the worst case.

The cultivation conditions for Rhizopus oryzae grown in synthetic medium and paper pulp spent sulfite liquor (SSL) were investigated to achieve high biomass and ethanol yields using shake flasks and bioreactors. The fungus assimilated the hexoses glucose, mannose and galactose, and the pentoses xylose and arabinose as well as acetic acid which are present in SSL. The assimilation of hexoses was faster than pentoses during cultivation in a synthetic medium. However, all sugars were assimilated concomitantly during growth in SSL supplemented with ammonium, magnesium, calcium, phosphate, sulfate and trace amounts of some other metal ions (SSL-S). The medium composition had an important influence on biomass yield. The highest biomass yields, viz. 0.18 and 0.43 g biomass/g sugar were obtained, when the cells were cultivated in shake flasks with a synthetic medium containing glucose as carbon and energy source and SSL-S, respectively. The corresponding yields in a bioreactor with more efficient aeration were 0.22 and 0.55 g/g. In addition to the biomass, ethanol, lactic acid, and glycerol were important extracellular metabolites of the cultivation with maximum yields of 0.37, 0.30 and 0.09 g/g, respectively. When the source of sugars in the medium was exhausted, the fungus consumed the metabolites produced, such that the liquid medium was depleted of potential oxidizable nutrients. In general, there was a direct competition between lactic acid and ethanol among the metabolites. Poor medium compositions and cultivation conditions resulted in higher yields of lactic acid, whereas the ethanol and biomass yields were higher in rich media. SSL-S supported good growth of mycelium and a high ethanol yield.

Here we developed a flow-based system resulting in improved performance of enzyme cascade-based biofuel cells. A paper-based biofuel cell with passive laminar flow was build to show the impact of flow on the performance of two different enzyme cascades – methanol and ethanol cascade. Both cascades demonstrated enhanced electrochemical output as a consequence of the decreased diffusion path of reaction intermediates identifying the intermediates diffusion in between enzymatic active sites as the rate limiting step in cascade operating systems.

The cultivation of Trametes versicolor for laccase production and cell growth were strongly dependent on experimental conditions namely physical and chemical parameters as well as nutrient availability and inducer stimulation. Biomass growth was compared for a rich medium and for a defined medium in two different temperatures. The best temperature was 28oC and the maximum specific growth rates were µ max = 0.083 h -1 for the rich medium and µ max = 0.043 h -1 for the defined medium. It was clearly shown that laccase production is not associated with cell growth, indicating that this ligninolytic enzyme must be produced in the defined medium by a secondary metabolism. In order to obtain laccase induction, addition of solid lignin, lignosulphonates, veratryl alcohol, xylidine and ethanol was tested at different concentrations. To optimise laccase activity, the combined effect of inducer addition and simultaneously glucose suppression was studied. The best result for laccase induction (1240 U/L) was obtained with solid lignin, a by-product of pulp and paper industry and the higher laccase activity attained (1583 U/L) was obtained with the combined effect of xylidine addition and glucose suppression.

An unexpected product was detected during simultaneous saccharification and co-fermentation (SSCF) of paper sludge using added commercial cellulase (Spezyme CP) by Saccharomyces cerevisiae RWB222, S. cerevisiae D5A, and Zymomonas mobilis 8b. Based on glycosyl composition analysis, linkage analysis and NMR analysis, the compound was identified as ethyl β-xylopyranoside (EXP). The carbon mass balance analysis showed up to 25% of xylan originally present in paper sludge was converted to EXP. EXP formation was found in simultaneous saccharification of beech wood xylan as well, and later proved to be produced by the Trichoderma reesei derived cellulase and hemicellulase mixture (Spezyme CP) during the course of xylan hydrolysis in the presence of ethanol, and its production increased with an increased concentration of ethanol, xylan, and T. reesei enzyme. Similar condensation reactions were also observed with other alcohols. These alcoholysis reactions were found to be reversible. Thermoanaerobacterium saccharolyticum was found to be able to degrade EXP.

Flexible sensors for the detection of ethanol at room temperature have been developed by depositing sensitive layers consisting of ITO nanopowder dispersed into poly-diallyldimethylammonium chloride (PDDAC), on the top of interdigitated electrodes based on conductive carbon nanotubes, previously printed onto flexible glossy paper substrates. The morphology and the microstructure of the interdigitated electrodes together with organic–inorganic sensing layer have been investigated by means of SEM, XRD and FT-IR. The electrical behaviour of the developed sensors at room temperature in response to low ethanol concentrations has been discussed as object of our research. The good ethanol sensing properties exhibited at room temperature by the composite film was supposed to originate from the electrostatic interaction occurring between the positively charged PDDAC polyelectrolyte and negatively charged ITO particles.

The spin trapping method combined with EPR spectroscopy has been employed by a number of laboratories to study alcohol-induced oxidative stress. Ethanol is converted to a free radical metabolite, the 1-hydroxyethyl radical in chemical reactions that generate hydroxyl radicals, in incubations of rat liver microsomes and in vivo. Furthermore, both acute and chronic ethanol administration leads to formation of radicals in the liver that are presumed to be products of lipid peroxidation. In general, overall radical production is enhanced by treatments known to be involved in alcoholic liver injury, such as chronic alcohol administration along with high levels of dietary fat.

Malondialdehyde and acetaldehyde react together with proteins in a synergistic manner and form hybrid protein adducts, designated as MAA adducts. MAA-protein adducts are composed of two major products whose structures and mechanism of formation have been elucidated. MAA adduct formation, especially in the liver, has been demonstrated in vivo during ethanol consumption. These protein adducts are capable of inducing a potent immune response, resulting in the generation of antibodies against both MAA epitopes, as well as against epitopes on the carrier protein. Chronic ethanol administration to rats results in significant circulating antibody titers against MAA-adducted proteins, and high anti-MAA titers have been associated with the severity of liver damage in humans with alcoholic liver disease. In vitro exposure of liver endothelial or hepatic stellate cells to MAA adducts induces a proinflammatory and profibrogenic response in these cells. Thus, during excessive ethanol consumption, ethanol oxidation and ethanol-induced oxidative stress result in the formation of acetaldehyde and malondialdehyde, respectively. These aldehydes can react together synergistically with proteins and generate MAA adducts, which are very immunogenic and possess proinflammatory and profibrogenic properties. By virtue of these potentially toxic effects, MAA adducts may play an important role in the pathogenesis of alcoholic liver injury.

The aim of this study was to characterize CYP2E1 degradation in vivo using PS-341, a potent proteasome inhibitor. Previously, only in vitro evidence showed that CYP2E1 induced by ethanol is degraded by the proteasome. Male Wistar rats were given ethanol intragastrically for 30 d. Ethanol was withdrawn at the same time that PS-341 was injected, 24 h before the rats were sacrificed. The liver proteasomal chymotrypsin-like activity (ChT-L) in rats fed ethanol was inhibited. After ethanol withdrawal, the proteasomal ChT-L activity returned to control levels. In the ethanol-withdrawn rats injected with PS-341, the ChT-L activity was significantly inhibited before withdrawal (p < .001). Ethanol treatment induced a 3-fold increase in CYP2E1 levels determined by Western blot. When ethanol was withdrawn, CYP2E1 decreased to control levels. In ethanol-withdrawn rats injected with PS-341, CYP2E1 remained at the induced level. These results show, for the first time, that the proteasome is responsible for ethanol-induced CYP2E1 degradation in vivo. Supported by NIH/NIAAA 8116.

A simple and sensitive fluorometric assay method for taurine (2-aminoethanesulfonic acid) has been developed. For the separation of taurine, high voltage paper electrophoresis subsequent to column chromatographic procedures was employed. Fluorescent product of taurine was yielded by spraying fluorescamine (4-phenylspiro [furan-2(3H), 1'-phthalan]-3, 3'-dione) and borate buffer on the paper, and the fluorescence was assayed spectro-fluorometrically after eluting with 50% ethanol. The linear relationship between the concentration of taurine and fluorescence developed was achieved over the concentration ranges of 0.5-10 nmoles, and the recoveries obtained were 90-100%. The specificity of this method for taurine was satisfactory and structural analogues involved in the metabolic pathway of taurine did not interfere with the assay. Examples for tissue levels of taurine in various organs of the rat as determined by this new method are also presented.

A simple quantitative procedure for sugar analysis of plant materials by paper chromatography with good reproducibility is described. The alcoholic extract was partially purified by a tertiary solvent system and ion exchange resins and then paper chromatography with the following solvent: water-saturated n -butanol, 95% ethanol and trichloroethylene (6:2:2, v/v). The sugars were selected by a guide strip technique, and eluted from the “cut-out squares” of paper with 3 ml distilled water by a micro-elution tube, here described. Theyb were determined by a micro-colorimetric method. The method permitted 72 determinations to be done at one time.

Ascending paper chromatographic methods for the separation of 2,4-dinitrophenylsulfide derivatives of mercaptans and mercapto-acids are presented. A solvent system composed of methanol as the stationary phase and heptane as the mobile phase gave satisfactory separations of the normal C 1 to C 7 mercaptan derivatives on What-man No. 1 paper. Two procedures were devised to separate the mercapto-acids derivatives. One method employed a solvent system of n -butanol:ethanol:water (20:5:25) with the water-rich layer as the stationary phase and the butanol-rich layer as the mobile phase. The solvent system for the second procedure was composed of equal parts of benzene and 1% acetic acid with the benzene-rich layer as the mobile phase in this case.

We describe a method for the assay of adenyl cyclase in whole tissue homogenates. Adenosine 3′:5′-cyclic monophosphate (cAMP) formed from α-32P-, 14C- or 3H-labeled adenosine 5′-triphosphate (ATP) substrate is isolated from all known ATP metabolites and an unknown metabolite by electrophoresis in 1% sodium borate for 40 min, followed by overnight descending chromatography in 95% ethanol:1 m ammonium acetate (70:30). The purity of the cAMP isolated is established by chromatographic techniques as well as by utilizing a purified cyclic nucleotide phosphodiesterase. The method described here also makes possible the measurement of phosphodiesterase activity in homogenates. It is rapid enough to allow routine assay of 180 samples per day, although the number of samples processed depends on the number of electrophoretic and chromatographic units available.

Ascending paper chromatography procedures are described for the separation of nitro and nitrato complexes of nitrosylruthenium in nitric acid solution. The elutants are methyl isopropyl ketone and dibutyl cellosolve. Tables of R/sub F/ for these elutants and each complex are given. (D.L.C.)

The influence of variations in temperature, presence and absence of buffers, and presence of other compounds in the applied solution on the R F values of 70 organic compounds has been evaluated in phenol-water and butanol-propionic acid-water systems. R F values of selected compounds in ethanol-acetic acid, butanol-ethanol-water, butanol-acetic acid-water, and ethanol are given. The production of methionine sulfoxide from methionine in phenol-water is discussed.

A two-reagent system, p -anisidine/ethanol followed by sodium periodate/acetone solution, is described for the detection and differentiation of sugars and sugar derivatives by their color reactions. The sensitivity of the method ranges from 1 to 20 μg, depending on the substances assayed. Polyols, sugar acids, 2-deoxy sugars, amino sugars and their acetyl derivatives, pentoses, and hexoses can be identified and characterized by this method.

Engineering microbes to produce value-added chemicals from C6/C5 sugars sometimes requires long biosynthetic pathways, which causes carbon loss due to involving multiple metabolic branch nodes, leading to a lower product yield. Using C2 feedstocks derived from gaseous, cellulosic, and plastic wastes could establish shorter biosynthetic pathways to produce some target chemicals, for example, acetyl-CoA-derived natural products. Utilizing these waste-derived feedstocks would also contribute to reducing the carbon footprint of the chemical industry. In this review, we highlighted the promising waste-processing technologies that could provide C2 feedstocks that are compatible with microbial fermentation. We also analyzed the recent metabolic engineering works in which the microorganisms/fermentation processes were modified/optimized to utilize acetate, ethanol, or ethylene glycol more efficiently.

Problems of stability were found for biomarkers of alcohol consumption: ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanols (PEths), and fatty acid ethyl esters (FAEEs) in whole blood. The purpose of this study was to establish a method for the determination of these four kinds of ethanol's non-oxidative metabolites in dried blood spots (DBS) by liquid chromatography tandem mass spectrometry (LC-MS/MS), and to evaluate their stability. In this method, 50 μL of human blood was spotted onto a filter paper for DBS analysis. Samples were extracted by methanol, reconstituted by 2-propanol, and injected into the LC-MS/MS system. Limits of detection were among 0.5–50 ng/mL, and deviations in accuracy and precision were all lower than 15% at three quality control levels. The stability of the four kinds of ethanol non-oxidative metabolites in DBS was investigated during a 90-day range under three temperatures, −20 °C, 4 °C, and 25 °C. EtG and EtS showed a high level of stability in DBS in the 90-day range, regardless of the temperature. FAEEs were unstable after three days. PEths showed stability within 15 days in postmortem DBS and 60 days in antemortem DBS, respectively, at all temperatures.

A general fluorescence procedure utilizing 0.5% and 1% solutions of quinine sulfate. 2H2O in absolute ethanol or 1% sulfosalicylic acid in acetone has been developed for the non-selective detection of a variety of organic compounds on paper chromatograms. When the developed and dried papers are dipped in the appropriate solution, dried and observed under ultraviolet light, organic compounds are visible against the background fluorescence of the paper. Inorganic or non-volatile developing solvents interfere with this procedure. Included in the list of compounds which were tested is the sapogenin lucernic acid which has heretofore defied detection on paper chromatograms.

Cassava processing wastes are suitable substrates for bioethanol production due to their high starch contents. However, these cassava wastes often contain various substances that inhibit cell growth and ethanol production. The abilities of multi-stress tolerant Saccharomyces cerevisiae LC269108 and Pichia kudriavzevii LC375240 to produce ethanol from cassava industrial wastes were evaluated. Analysis of cassava peels and pulps from two cassava starch factories showed that the cassava peels contained high cyanide (KCN) concentrations while cassava pulps contained high concentrations of sodium dodecyl benzene sulfonate (SDBS) used to improve starch extraction in the factory. Although S. cerevisiae LC269108 produced ethanol efficiently from the cassava peel, its growth and ethanol production were inhibited in the cassava pulp. On the other hand, the growth and ethanol production by P. kudriavzevii LC375240 were inhibited slightly in the cassava peel but not in the cassava pulp. Based on the intracellular pyruvate concentrations in the presence of KCN and SDBS, it was inferred that their inhibition site is on the pyruvate synthetic pathways. With a co-culture of the two strains, ethanol production from the cassava peel increased significantly but there was no significant improvement in ethanol production from the cassava pulp.

The retention of aniline derivatives, sulfonamides, and several N-heterocyclics on paper impregnated with a strong acidic H + -form resin was investigated. Solvent systems used were ethanol, acetonitrile, dimethylformamide, dimethylsulfoxide, pyridine, and butylamine mixed with water. Either aprotic, amphiprotic, basic, or acidic developing mixtures can be used for separations. The retention data can be correlated, in general, to the p K a values for the weak bases and to column and batch retention data obtained with strongly acidic cation resin.

Lignin valorisation into materials such as resins is essential to increase the value obtained from biomass. However, biomass recalcitrance limits the selective isolation of lignin for economic gains. This study developed a new process for fractionating alkaline treated mango seed husk into high purity lignin and cellulose-rich pulp, using high shear homogenization-organosolv (HSHO) process. The HSHO process conditions (ethanol concentration (50-70%), temperature (130-150 °C) and homogenizing time (10-20 min)) were optimized using response surface methodology to maximize the solubilised lignin with high purity while obtaining a fibrillated cellulose-rich pulp. Optimum process conditions of 60% ethanol, 148.41 °C, and 15 min homogenization, yielded 70.23% lignin of 96.18% purity, higher than those of the non-assisted process (68.58% and 94.74%, respectively). Nuclear magnetic resonance spectroscopy showed syringyl and guaiacyl lignin units with a molecular weight of 3247 g/mol and thermal degradation temperature of 298 °C. Sulphur and nitrogen contents in the resulting lignin were lower than 0.15%. Fibrillated cellulose pulp with diameters of1-10 μm were obtained. This study has established the proficiency of an HSHO process for biomass fractionation and more so, for the extraction of lignin with90% purity suitable for varied applications.

A paper chromatographic method has been developed for the separation of the six platinum metals. The metals are first separated in two groups by distilling off osmium and ruthenium as tetroxides and absorbing them in caustic alkali. The two mixtures are then separated chromatographically employing a mixture of n -butanol, ethanol, hydrochloric acid, and water in the proportion of 60:10:10:20 by volume as the developing solvent.

A study is described of the chromatography on paper of some alkylhydrazines and some monoand diacylhydrazines by both the circular and ascending methods. For the acylhydrazines the solvent system preferred was the upper layer from isoamyl alcohol, acetic acid, and water in the volume proportions of 10:1.5 :10 respectively. The upper layer from a mixture of n -butyl alcohol, acetic acid, and water in the volume proportions of 4 : 1 : 5 respectively, was preferred for the alkylhydrazines. For detection of the hydrazine derivatives on the chromatogram a spray of p -dimethylaminobenzaldehyde hydrochloride in ethanol (Ehrlich's reagent) was most satisfactory, although ammoniacal silver nitrate or ninhydrin could be used in some cases.

Palm biodiesel-ethanol blends could be the better automotive fuel over palm biodiesel alone owing to the high oxygen content and high volatility of ethanol. The evaporation and burning characteristics of palm biodiesel with 10%, 20%, and 30% ethanol by volume concentration respectively, were investigated by using the droplet experiment. Rapid bubble growth and bubble explosions during the heating stage were observed to be substantial for the palm biodiesel droplet added with 30% volume of ethanol (BE30). Adding ethanol in palm biodiesel increased the ignition delay and burn-rate constant by up to 38.6% and 23.2%, respectively whereas, the evaporation duration and burning period decreased by up to 20.6% and 22.5%, respectively. Overall, the relatively shorter evaporation duration and higher burn-rate found for the BE30 droplet could promote high thermal efficiency and has the potential to achieve ultra-low carbon monoxide (CO) and unburned hydrocarbons (UHCs) emissions in diesel and gas turbine engines.

Life cycle assessments of fuel bioethanol produced in Uruguay from liquid hot water pretreated switchgrass were performed for two scenarios: the production of ethanol and electricity and a biorefinery producing ethanol, electricity, furfural, acetic, and formic acid. Experimental data and material and energy balances derived from the simulations were used to analyze the global warming potential of fuel bioethanol. Greenhouse gas (GHG) emissions in both scenarios were lower than the reference emissions for fossil fuel. The biorefinery scenario had a better performance in terms of environmental impacts, and the biofuel produced met GHG reduction requirements. Switchgrass composition, enzyme dosage, fermentation and hydrolysis efficiency, and solids content had a significant effect on environmental performance, with enzyme use being the most significant factor. A multi-objective optimization was used to find the values of solid content and enzyme dose that minimize GHG emissions and the minimum ethanol selling price (MESP). An enzyme dosage of 37 mgprotein/gglucan and solids content of 21% w/w were the optimal conditions for bioethanol production with an associated GHG value of 2 ± 5 g CO2 eq./MJethanol and a MESP of $0.84/L.

Industrial production processes will produce a large amount of inorganic carbon (CO/CO2), which can be used as a carbon substrate for gaseous fermentation to reduce environmental pollution and promote the development of renewable energy. To solve the bottleneck that the strains used for the gaseous fermentation cannot adapt well to the mixed gas (CO, CO2 and H2) and the low ethanol production, the mixed gas was aerated continuously into the culture with rabbit faeces to enrich ethanol-producing strains. After CO/CO2 domestication, the structure of the microbial community changed significantly. Blautia, which can use CO/CO2 to produce ethanol, became the dominant bacteria, and the relative abundance significantly increased by 5.4 times to 41.1%. What's more, 14.07% of CO/CO2 was converted to organic carbon through fermentation by the domesticated bacteria, and the ethanol concentration reached 1.41 g L−1. The results indicated that with the continuous CO/CO2 domestication, the bacteria could adapt the syngas better and showed an improved fermentation performance.

Decreasing carbon dioxide (CO2) emission by converting to higher-valued product has become of interest. Hydrogen (H2) is an important feedstock required in thermochemical conversion of CO2 to chemicals such as methanol. The cost and availability of H2 affect the cost of CO2 conversion. This study is focused on the process simulation of H2 production from ethanol feedstock. Steam reforming of ethanol is compared with dehydrogenation of ethanol to H2 with valued products including ethyl acetate and acetaldehyde. Form this study, steam reforming of ethanol presents the lowest cost of H2 production at 1.58 USD/kg H2 while dehydrogenation of ethanol presents the cost at 3.24 and 1.97 USD/kg H2, respectively. Although presenting the lowest cost, steam reforming of ethanol provides a net positive CO2 emission in the overall CO2 conversion to methanol process. In contrast, ethanol dehydrogenation to H2 and byproducts, ethyl acetate and acetaldehyde, promotes a net negative CO2 emission of −819.20 kg/ton methanol and −5.42 kg/ton methanol, respectively. The results present a decreasing CO2 emission with an increasing cost of H2 production.

Unwanted agricultural waste is largely comprised of lignocellulosic substrate which could be transformed into sugars. The production of bioethanol from garbage manifested an agreeable proposal towards waste management as well as energy causation. The goal of this work is to optimize parameters for generation of bioethanol through fermentation by different yeast strains while Saccharomyces cerevisiae used as standard strain. The low cost fermentable sugars from pomegranate peels waste (PPW) were obtained by hydrolysis with HNO3 (1 to 5%). The optimum levels of hydrolysis time and temperature were elucidated via RSM (CCD) ranging from 30 to 60 min and 50 to 100 °C respectively. The result shows that optimum values (g/L) for reducing sugars was 61.45 ± 0.01 while for total carbohydrates was 236 ± 0.01. These values were found when PPW was hydrolyzed with 3% HNO3, at 75 °C for one hour. The hydrolyzates obtained from the dilute HNO3 pretreated PPW yielded a maximum of 0.43 ± 0.04, 0.41 ± 0.03 g ethanol per g of reducing sugars by both Metchnikowia sp. Y31 and M. cibodasensis Y34 at day 7 of ethanologenic experiment. The current study exhibited that by fermentation of dilute HNO3 hydrolyzates of PPW could develop copious amount of ethanol by optimized conditions.

Carbon loss in the form of CO2 is an intrinsic and persistent challenge faced during conventional and advanced biofuel production from biomass feedstocks. Current mechanisms for increasing carbon conservation typically require the provision of reduced co-substrates as additional reducing equivalents. This need can be circumvented, however, by exploiting the natural heterogeneity of lignocellulosic sugars mixtures and strategically using specific fractions to drive complementary CO2 emitting vs. CO2 fixing pathways. As a demonstration of concept, a coculture-coproduction system was developed by pairing two catabolically orthogonal Escherichia coli strains; one converting glucose to ethanol (G2E) and the other xylose to succinate (X2S). 13C-labeling studies reveled that G2E + X2S cocultures were capable of recycling 24% of all evolved CO2 and achieved a carbon conservation efficiency of 77%; significantly higher than the 64% achieved when all sugars are instead converted to just ethanol. In addition to CO2 exchange, the latent exchange of pyruvate between strains was discovered, along with significant carbon rearrangement within X2S.