Effects of Spray Drying on Physicochemical Properties of Chitosan Acid Salts

The effects of spray-drying process and acidic solvent system on physicochemical properties of chitosan salts were investigated. Chitosan used in spray dryings was obtained by deacetylation of chitin from lobster (Panulirus argus) origin. The chitosan acid salts were prepared in a laboratory-scale spray drier, and organic acetic acid, lactic acid, and citric acid were used as solvents in the process. The physicochemical properties of chitosan salts were investigated by means of solid-state CP-MAS ¹³C nuclear magnetic resonance (NMR), X-ray powder diffraction (XRPD), differential scanning calorimetry, and Fourier transform infrared spectrometry (FTIR) and near-infrared spectroscopy. The morphology of spray-dried chitosan acid salts showed tendency toward higher sphericity when higher temperatures in a spray-drying process were applied. Analysis by XRPD indicated that all chitosan acid salts studied were amorphous solids. Solid-state ¹³C NMR spectra revealed the evidence of the partial conversion of chitosan acetate to chitin and also conversion to acetyl amide form which appears to be dependent on the spray-drying process. The FTIR spectra suggested that the organic acids applied in spray drying may interact with chitosan at the position of amino groups to form chitosan salts. With all three chitosan acid salts, the FTIR bands at 1,597 and 1,615 cm⁻¹ were diminished suggesting that –NH groups are protonated. The FTIR spectra of all chitosan acid salts exhibited ammonium and carboxylate bands at 1,630 and 1,556 cm⁻¹, respectively. In conclusion, spray drying is a potential method of preparing acid salts from chitosan obtained by deacetylation of chitin from lobster (P. argus) origin.     

Spectrophotometric assay of long-chain unsaturated and hydroxy fatty acids in concentrated sulfuric acid

A spectrophotometric method has been developed for the determination of long-chain unsaturated and hydroxy fatty acids in concentrated sulfuric acid. The assay is based on the absorbance produced in the 290 to 300-nm range from their reaction with sulfuric acid at 100°C. α,β-Unsaturated aliphatic acids give absorption bands at 235–240 nm and thus can be easily differentiated from unsaturated fatty acids having the double bond(s) at positions not conjugated with the carboxyl group. A certain minimum chain length is required for full development of the absorption band at 300 nm. Position and intensity of the so-formed absorption band is independent on the position and number of the double bonds or hydroxyl groups. Carboxyl groups are not essential, as unsaturated hydrocarbons and higher alcohols likewise react with sulfuric acid to produce the absorbing species at 300 nm, providing a minimum chain length of 5 carbon atoms is present. The nature of the absorbing species at 300 nm is discussed.     

Tetradecane 1,14-Dicarboxylic Acid Production from n-Hexadecane by Candida cloacae

The better condition of cultivation for tetradecane 1,14-dicarboxylic acid (DC-16) production from n-hexadecane (n-C₁₆) by Candida cloacae MR-12 was investigated by using acetic acid as carbon source for the growth. In general, the condition suitable for the growth was also favorable for the production of DC-16. The change of pH during cultivation, the use of NaOH solution as pH controlling agent after pH-change and the addition of antifoam stimulated the production of DC-16.

Under the optimum conditions where the culture medium contained 15% (v/v) n-C₁₆, 1.4% (w/v) acetic acid, inorganic salts and growth factors, and pH was changed from 6.5 to 7.75 at 16 hr after the inoculation, the highest level of DC-16 production was attained after about 72 hr cultivation and the amount of the product accumulated was 61.5 g per liter of the medium.

When a mixture of various n-alkanes was used as starting material, DCs corresponding to the respective n-alkanes were produced as mixture.     

Biotechnological Applications of Acetic Acid Bacteria

The acetic acid bacteria (AAB) have important roles in food and beverage production, as well as in the bioproduction of industrial chemicals. In recent years, there have been major advances in understanding their taxonomy, molecular biology, and physiology, and in methods for their isolation and identification. AAB are obligate aerobes that oxidize sugars, sugar alcohols, and ethanol with the production of acetic acid as the major end product. This special type of metabolism differentiates them from all other bacteria. Recently, the AAB taxonomy has been strongly rearranged as new techniques using 16S rRNA sequence analysis have been introduced. Currently, the AAB are classified in ten genera in the family Acetobacteriaceae. AAB can not only play a positive role in the production of selected foods and beverages, but they can also spoil other foods and beverages. AAB occur in sugar- and alcohol-enriched environments. The difficulty of cultivation of AAB on semisolid media in the past resulted in poor knowledge of the species present in industrial processes. The first step of acetic acid production is the conversion of ethanol from a carbohydrate carried out by yeasts, and the second step is the oxidation of ethanol to acetic acid carried out by AAB. Vinegar is traditionally the product of acetous fermentation of natural alcoholic substrates. Depending on the substrate, vinegars can be classified as fruit, starch, or spirit substrate vinegars. Although a variety of bacteria can produce acetic acid, mostly members of Acetobacter, Gluconacetobacter, and Gluconobacter are used commercially. Industrial vinegar manufacturing processes fall into three main categories: slow processes, quick processes, and submerged processes. AAB also play an important role in cocoa production, which represents a significant means of income for some countries. Microbial cellulose, produced by AAB, possesses some excellent physical properties and has potential for many applications. Other products of biotransformations by AAB or their enzymes include 2-keto-L-gulonic acid, which is used for the production of vitamin C; D-tagatose, which is used as a bulking agent in food and a noncalorific sweetener; and shikimate, which is a key intermediate for a large number of antibiotics. Recently, for the first time, a pathogenic acetic acid bacterium was described, representing the newest and tenth genus of AAB.     

Fatty Acid Metabolism in Microorganisms

During the investigation on the metabolism of azelaic acid by Micrococcus sp., it was found that the bacterium produced a large amount of keto acid (α-ketoglutaric acid) under the restricted condition for nitrogen source. The acid was identified as α-ketoglutaric acid by physico-chemical and biological methods. The mechanism of the production of α-ketoglutaric acid from azelaic acid was investigated. From the result, it was suggested that α-ketoglutaric acid production proceeded thrpugh the further oxidation of acetic acid produced from azelaic acid and that the production might be functioned by TCA cycle enzymes of the bacterium. Similarly, α-ketoglutaric acid was found to be produced remarkably from other various fatty acids.     

STIMULATION OF FUNDULUS BY HYDROCHLORIC AND FATTY ACIDS IN FRESH WATER,AND BY FATTY ACIDS,MINERAL ACIDS,AND THE SODIUM SALTS OF MINERAL ACIDS IN SEA WATER

1. Fundulus heteroclitus was found to be a reliable experimental animal for studies on chemical stimulation in either fresh or sea water. 2. The response of Fundulus to hydrochloric, acetic, propionic, butyric, valeric, and caproic acids was determined in fresh water, while the same acids plus sulfuric and nitric, as well as the sodium salts of the mineral acids, were tested in sea water. 3. Stimulation of Fundulus by hydrochloric acid in fresh water is correlated with the effective hydrogen ion concentration. Stimulation by the n-aliphatic acids in the same environment is correlated with two factors, the effective hydrogen ion concentration and the potential of the non-polar group in the molecule. However, as the number of CH₂ groups increases the stimulating effect increases by smaller and smaller amounts, approaching a maximum value. 4. Stimulation of Fundulus by hydrochloric, sulfuric, and nitric acids in sea water is correlated with the forces of primary valence which in turn are correlated with the change in hydrogen ion concentration of the sea water. The n-aliphatic acids increase in stimulating efficiency in sea water as the length of the carbon chain increases, but a limiting value is not reached as soon as in fresh water. 5. Only a slight difference in stimulation by hydrochloric acid is found in sea water and in fresh water. However, there is a significant difference in stimulation by the fatty acids in fresh and in sea water, which is partly explained by the different buffering capacities of the two media. It is to be noted that in the same environment two different fish, Fundulus and Eupomotis, give different results, while the same fish (Fundulus) in two different environments responds similarly to mineral acids but differently to fatty acids. These results illustrate that stimulation is a function of the interaction between environment and receptors, and that each is important in determining the response. 6. Stimulation by sodium chloride, nitrate, and sulfate is correlated with equivalent concentrations of the salts added to sea water, or with the forces of primary valence. Although the threshold for stimulation by the salts is considerably higher than for the acids, the efficiency of stimulation by the salts is greater.     

The estimation of acid glycosaminoglycan-Alcian blue complexes eluted from electrophoretic strips

The present report describes a new and simple method for the detection and quantitative determination of pentoses and hexuronic acids either separately or in a mixture. The method involves treatment of the sample with sulfuric acid followed by the addition of harmine and then cysteine. Hexuronic acids produce a pink color with harmine, while pentoses react only after the addition of cysteine. Hexosamines, methylpentoses, and moderate amounts of hexoses do not interfere with these determinations.The method is simple, accurate, and allows the determination of small amounts of pentoses in the presence of larger amounts of hexuronic acids and other sugars.     

Ion-pair extraction of bile acids with Lipidex gel

A new method for the extraction of bile acids from aqueous solutions, urine, plasma, and bile is described. A buffered solution of decyltrimethylammonium bromide is added to the sample to give a 0.03 m concentration of the counter-ion. The mixture is passed through a bed of Lipidex 1000, which is then washed with the buffered solution of counter-ion followed by water. The decyltrimethylammonium salts of bile acids are sorbed by the Lipidex and are eluted with methanol. Recoveries of unconjugated, taurine- and glycine-conjugated, sulfated, and glucuronidated bile acids are close to 100%. Unconjugated bile acids can also be quantitatively extracted from aqueous solutions and urine after acidification with acetic acid.     

Synchronous fluorescence determination of ferulic acid with Ce(IV) and sodium tripolyphosphate

In this study, a synchronous fluorescence detection method for ferulic acid (FA) is proposed based on a redox reaction between FA and Ce(IV) sulfate in dilute sulfuric acid medium at room temperature. It was found that FA could reduce Ce(IV) to Ce(III) in acidic medium, and sodium tripolyphosphate could further enhance the intrinsic fluorescence of the Ce(III) produced. The enhanced extent of synchronous fluorescence intensity was in proportion to the concentration of FA over the range 3.0 × 10^‐8 to 1.0 × 10^‐5 mol/L. The corresponding limit of determination (S/N = 3) was 1.3 × 10^‐8 mol/L. The proposed method was applied to the determination of sodium ferulate for injection sample with satisfactory results. Copyright © 2013 John Wiley & Sons, Ltd.     

Succinic Acid Production with Actinobacillus succinogenes ZT-130 in the Presence of Succinic Acid

Glucose fermentation with Actinobacillus succinogenes was carried out at different initial concentrations of succinic acid (SA₀) to determine its effect on growth and on the production of succinic acid itself. The specific rates of biomass production, succinic, formic and acetic acids decreased with SA₀ (0–40 g/l). The partially dissociated form of succinic acid had a higher effect on cell growth and production of succinic acid as compared to the non-dissociated forms of the acids, a fact that has not been reported until now. Cell growth fitted the Jerusalimski model, and the Aiba–Shoda model was suitable for quantification of the inhibition for the production of succinic acid. The growth inhibition constants K _is and K _ip and their summatory were consistent with the experimental values obtained, i.e., 22 g/l for the produced acids and 38 g/l for total acids that were the limits at which the biomass formation ceased.     

Effects of different pretreatment strategies on corn stalk acidogenic fermentation using a microbial consortium

The effects of sulfuric acid, acetic acid, aqueous ammonia, sodium hydroxide, and steam explosion pretreatments of corn stalk on organic acid production by a microbial consortium, MC1, were determined. Steam explosion resulted in a substrate that was most favorable for microbial growth and organic acid productions. The total amounts of organic acids produced by MC1 on steam exploded, sodium hydroxide, sulfuric acid, acetic acid, and aqueous ammonia pretreated corn stalk were 2.99, 2.74, 1.96, 1.45, and 2.21 g/l, respectively after 3 days of fermentation at 50 °C. The most prominent organic products during fermentation of steam-exploded corn stalks were formic (0.86 g/l), acetic (0.59 g/l), propanoic (0.27 g/l), butanoic (0.62 g/l), and lactic acid (0.64 g/l) after 3 days of fermentation; ethanol (0.18 g/l), ethanediol (0.68 g/l), and glycerin (3.06 g/l) were also produced. These compounds would be suitable substrates for conversion to methane by anaerobic digestion.     

The Colorimetric Determination of Isobutyl and Isoamyl Alcohol in Fusel Oil Separately

The amounts of isobutyl and isoamyl alcohol which are the main constituents of fusel oil are determined fractionally. Isobutyl and isoamyl alcohol give different colors by treatment with p-dimethylaminobenzaldehyde in sulfuric acid. Using this color difference the amount of each of these alcohols in fusel oil is determined colorimetrically. These alcohols arise during alcoholic fermentation and are concentrated in the distillate by distillation, and affect the flavor of distilled spirits, such as brandy and whisky, i.e., butyl alcohol is buttery and amyl alcohol is aromatic. So determination of the amounts of these alcohols fractionally in these spirits furnishes a useful criterion for evaluating the character of these spirits.     

Conformational Behavior of Chitosan in the Acetate Salt: An X-Ray Study

A well-defined X-ray fiber pattern of chitosan acetate was obtained by immersing a tendon chitosan, prepared from a crab tendon chitin by a solid-state N-deacetylation, in an aqueous acetic acid-isopropanol solution at 110°C. This pattern was very similar to that of chitosan salts with some inorganic acids, such as HF, HCl, and H₂SO₄, in which chitosan chains form an 8/5 helix, indicating that chitosan acetate also take up this conformation. This information may give an influential clue to the chitosan conformation in the aqueous acetic acid solution, the most popular solvent for chitosan. However, after one month of storage of the chitosan acetate, the fiber pattern, the density and its IR spectrum changed to those of the anhydrous polymorph of chitosan, suggesting that the acetic acid was removed accompanied with water molecules from the crystal during storage and that the polymorph can be obtained not only by annealing chitosan, but also through the chitosan acetate.     

An Interpretation of the Mass Spectra of Salinomycin and Its Derivatives

Salinomycin is a new polyether antibiotic elaborated by Streptomyces albus. An interpretation of the mass spectra of salinomycin and its derivatives has been achieved with the aid of high resolution mass spectrometry along with measurement of the metastable ions. Interesting aspects of the mass spectral data of these compounds are that the characteristic cleavage occurred at the ring C of the tricyclic spiroketal system, and that the sodium salts of salinomycin itself and some derivatives thereof are volatile enough to give distinct spectra which showed fragmentations comparable with those of the free acid and the methyl ester.

An interpretation of the principal fragment ions and their formation mechanism for salinomycin and its derivatives is discussed.     

Weathering of calcite,pyrite, and sulfur by Thermothrix thiopara in a thermal spring

The mineral weathering capabilities of Thermothrix thiopara were investigated by scanning electron microscopy and energy dispersive X‐ray analysis. Thermothrix thiopara is an extremely thermophilic, sulfur‐oxidizing bacterium that grows in a thermal spring whose principal minerals are calcium carbonate, pyrite, and sulfur. Crystals of these minerals were incubated in situ for periods up to eight days, removed, and examined. Results indicated that T. thiopara is partially responsible for weathering calcium carbonate by the production of sulfuric acid, thereby contributing to the formation of a porous tufa mound. Examination of ultravioletirradiated control crystals indicated that the sulfuric acid produced by T. thiopara caused solubilization of calcium carbonate even in the absence of direct bacterial colonization. Pyrite and sulfur were not visibly weathered, but instead were coated with calcium carbonate precipitate. During eight days incubation, growth forms of T. thiopara colonizing the minerals progressed from unicells to filaments to nets of filaments. Bacteria other than T. thiopara appeared after eight days, indicating an increased diversity.     

A simple and specific determination of glycine in biological samples

A simple specific assay was developed for the determination of glycine in a solution containing other amino acids. Hippuric acid was obtained after reacting glycine with benzoyl chloride and was extracted with ethyl acetate. It was then reacted with acetic anhydride, p-dimethylaminobenzaldehyde, and pyridine for color development. The amount of glycine (1 to 100 μg) in the original solution could be determined by measuring the absorbance (458 nm) of this chromogen. This procedure was applied on an amino acid mixture, urine, serum, blood, and liver homogenate.     

Rapid chromatographic method to determine polyamines in urine and whole blood

A procedure is described for the rapid determination of putrescine, spermine and spermidine in urine and whole blood. The samples are hydrolyzed with barium hydroxide and are neutralized with sulfuric acid. The polyamines are concentrated and separated from amino acids on a small bed of ion-exchange resin that then serves to load the samples on a two-channel, automated ion-exchange chromatography apparatus. As many as 100 samples can be analyzed in a 24-h period. The method has been shown to be applicable to the analysis of urine and whole blood samples, but further development is needed for application to serum samples.     

Acetic Acid Production by an Electrodialysis Fermentation Method with a Computerized Control System

In acetic acid fermentation by Acetobacter aceti, the acetic acid produced inhibits the production of acetic acid by this microorganism. To alleviate this inhibitory effect, we developed an electrodialysis fermentation method such that acetic acid is continuously removed from the broth. The fermentation unit has a computerized system for the control of the pH and the concentration of ethanol in the fermentation broth. The electrodialysis fermentation system resulted in improved cell growth and higher productivity over an extended period; the productivity exceeded that from non-pH-controlled fermentation. During electrodialysis fermentation in our system, 97.6 g of acetic acid was produced from 86.0 g of ethanol; the amount of acetic acid was about 2.4 times greater than that produced by non-pH-controlled fermentation (40.1 g of acetic acid produced from 33.8 g of ethanol). Maximum productivity of electrodialysis fermentation in our system was 2.13 g/h, a rate which was 1.35 times higher than that of non-pH-controlled fermentation (1.58 g/h).     

Determination of volatile fatty acids in plasma after ethanolic extraction

1. A new rapid micro-method for measuring plasma volatile fatty acids is described. The volatile fatty acids are extracted from plasma with ethanol in the presence of a known quantity of internal standard (sodium isobutyrate). After evaporation of the ethanolic solution of the sodium salts, the residue is dissolved in a dilute solution of orthophosphoric acid to permit analysis by g.l.c. 2. A technique of g.l.c. analysis is described which permits the separation of all the volatile fatty acids from the other plasma constituents at temperatures below 100 degrees C in 5 min. 3. Steam-distillation techniques are unsatisfactory when the acetic acid concentrations in the plasma are below 0.2mm. Heating of a number of plasma constituents in acid conditions gives rise to acetic acid. 4. The binding of volatile fatty acids to plasma proteins was studied; this binding is negligible for acetic acid, but increases with the length of the fatty acid carbon chain. 5. The limits of use of the method and the physiological implications are discussed.     

Feulgen Hydrolysis with Perchloric Acid

In tissue fixed with Carnoy's acetic alcohol (1:3), the hydrochloric acid hydrolysis performed as part of the Feulgen reaction is optimal for only a very short period of time. When 10% perchloric acid is used as the hydrolytic agent, the same color maximum is obtained, and the optimal hydrolysis time at 25°C. extends from 12 hours to 24 hours. During this time the intensity of color does not change. The events which take place during the period of suboptimal hydrolysis are the same as diose which take place during the corresponding period of hydrochloric acid hydrolysis. Part of the decrease in ultraviolet extinction of nuclei during the first 12 hours is due to the splitting off of purine bases from the desoxyribose nucleic acid. This is consistent with the increase in the amount of Feulgen dye bound by nuclei during this period of time. Between 12 hours and 24 hours no ultraviolet absorbing material is lost from nuclei, which is consistent with the fact that during this time the Feulgen color produced remains at a maximum.