Reactivity of ferulic acid and its derivatives toward hydrogen peroxide and peracetic acid.

The reactions of ferulic acid and its derivatives with hydrogen peroxide and peracetic acid in lignin-retaining bleaching conditions have been investigated to determine their susceptibility to oxidative degradation. The conjugated side chain of ferulic acid and its etherified or esterified derivative was shown to be fairly stable, especially to hydrogen peroxide. The major reaction was trans-cis isomerization that possibly involved a radical mechanism but did not cause bond cleavage. The peracetic acid reaction increased the rate of trans-cis isomerization and was also accompanied by a minor cleavage of the side chain. Esterification did not have a substantial effect on the reactivity of ferulic acid, but 4-O-etherification significantly stabilized it against these two oxidants. By contrast, aldehyde substitution tremendously enhanced the susceptibility of the cinnamyl side chain to oxidative degradation, as evidenced by an intensive degradation of coniferaldehyde.     

Reactivity of human salivary proteins families toward food polyphenols

Tannins are well-known food polyphenols that interact with proteins, namely, salivary proteins. This interaction is an important factor in relation to their bioavailability and is considered the basis of several important properties of tannins, namely, the development of astringency. It has been generally accepted that astringency is due to the tannin-induced complexation and/or precipitation of salivary proline-rich proteins (PRPs) in the oral cavity. However, this complexation is thought to provide protection against dietary tannins. Neverthless, there is no concrete evidence and agreement about which PRP families (acidic, basic, and glycosylated) are responsible for the interaction with condensed tannins. In the present work, human saliva was isolated, and the proteins existing in saliva were characterized by chromatographic and proteomic approaches (HPLC-DAD, ESI-MS, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and MALDI-TOF). These approaches were also adapted to study the affinity of the different families of salivary proteins to condensed tannins by the interaction of saliva with grape seed procyanidins. The results obtained when all the main families of salivary proteins are present in a competitive assay, like in the oral cavity, demonstrate that condensed tannins interact first with acidic PRPs and statherin and thereafter with histatins, glycosylated PRPs, and bPRPs.     

Reactivity of certain organophosphorus insecticides toward hydroxyl radicals at elevated air temperatures

Methods were developed to determine OH reaction rates for medium-weight organophosphorus pesticides in the gas phase. A 57-L chamber was constructed that utilized xenon arc irradiation (>290 nm) to photolyze the OH precursor, methyl nitrite. Experiments were performed at elevated temperatures ranging from 60 to 80 degrees C to minimize wall sorption. Solid-phase microextraction (SPME) sampling of the gas phase was employed to assess the simultaneous rate of loss of the pesticides in relation to the rate of loss of two reference substances with known OH rate constants. An internal gas-phase standard (hexachlorobenzene), relatively stable to hydroxyl radicals, was used to assess other processes, which included dilution and wall sorption. The relative reaction rates of the organophosphorus insecticides, diazinon and chlorpyrifos, when compared to reference compounds, were unaffected by various air temperatures between 60 and 80 degrees C. Although both insecticides were expected to react at similar rates on the basis of structural activity model predictions, diazinon reacted 3 times more rapidly than chlorpyrifos and gave estimated environmental OH half-lives of 0.5 and 1.4 h, respectively. The degree of sorption onto the chamber walls was minimal and similar for each of the compounds examined. Experimental gas-phase determinations at elevated temperatures may provide important information that can be used when in the assessment of the potential of airborne pesticide risks to nontarget and ecologically sensitive areas.     

Interaction of the 2,4-dichlorophenoxyacetic acid herbicide with soil organic matter moieties: a theoretical study

The determination of the structure of humic substances from soils and natural waters is an intriguing problem in soil science. Humic substances consist of molecules covering a broad distribution of molecular size and involving different functional groups. Taking this into account, we have chosen smaller model systems with functional groups typically present in humic substances. We investigated theoretically, by quantum chemical calculations, the environmental effects on the complexes formed from the interaction of 2,4‐dichlorophenoxyacetic acid and its anion with acetaldehyde, methanol, methylamine, protonated methylamine, acetic acid and water. The important case of a cation bridge mechanism, with Ca²⁺ as the bridging cation, is also included into the set of model reactions. It is found that this cation bridge belongs to one of the most stable mechanisms of fixation of organic compounds in soils. According to our calculations the hydroxyl group forms the most stable complexes with 2,4‐D in a polar solvent environment.     

Kinetics of acid hydrolysis of water-soluble spruce O-acetyl galactoglucomannans

Water-soluble O-acetyl galactoglucomannan (GGM) is a softwood-derived polysaccharide, which can be extracted on an industrial scale from wood or mechanical pulping waters and now is available in kilogram scale for research and development of value-added products. To develop applications of GGM, information is needed on its stability in acidic conditions. The kinetics of acid hydrolysis of GGM was studied at temperatures up to 90 degrees C in the pH range of 1-3. Molar mass and molar mass distribution were determined using size exclusion chromatography with multiangle laser light scattering and refractive index detection. The molar mass of GGM decreased considerably with treatment time at temperatures above 70 degrees C and pH below 2. The molar mass distribution broadened with hydrolysis time. A first-order kinetic model was found to match the acid hydrolysis. The reaction rate constants at various pH values and temperatures were calculated on the basis of the first-order kinetic model. Furthermore, the activation energy, E, was obtained from the Arrhenius plot. The activation energy E was 150 kJ mol (-1) for acid hydrolysis of spruce GGM. The apparent rate constant during acid hydrolysis increased by a factor of 10 with a decrease in pH by 1 unit, regardless of temperature. In addition, gas chromatography and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were applied to study the released GGM monomers and oligomers.     

Distribution of a new rosmarinic acid derivative in Eryngium alpinum L. and other Apiaceae

The roots of Eryngium alpinum L. (Apiaceae) demonstrated radical scavenging properties toward the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical in a TLC autographic assay. Isolation of the bioactive compounds allowed the identification of R-(+)-3'-O-beta-D-glucopyranosyl rosmarinic acid, a new rosmarinic acid derivative. The quantitative determination of the antioxidant capacity of the compound by chemoluminescence demonstrated half the activity of R-(+)-rosmarinic acid. To find another source richer in this compound, a chemotaxonomic study was conducted. The higher content was found in the aerial parts of Sanicula europeae L., also belonging to the Saniculoideae subfamily. Although present in most of the Eryngium species, this compound was not detected in Imperatoria ostruthium L., Pimpinella peregrina L., and Levisticum officinalis L. species from the Apioideae subfamily and Hydrocotyle asiatica L. from the Hydrocotyloideae subfamily. The results indicate that the new derivative R-(+)-3'-O-beta-D-glucopyranosyl rosmarinic acid is a potential chemotaxonomic marker of the Saniculoideae subfamily.     

Chemical structures of corn stover and its residue after dilute acid prehydrolysis and enzymatic hydrolysis: insight into factors limiting enzymatic hydrolysis

Advanced solid-state NMR techniques and wet chemical analyses were applied to investigate untreated corn stover (UCS) and its residues after dilute acid prehydrolysis (DAP) and enzymatic hydrolysis (RES) to provide evidence for the limitations to the effectiveness of enzyme hydrolysis. Advanced solid-state NMR spectral-editing techniques as well as 1H-13C two-dimensional heteronuclear correlation NMR (2D HETCOR) were employed. Our results indicated that dilute acid prehydrolysis selectively removed amorphous carbohydrates, increased aromatic CH/other protonated -C═C- and enriched alkyl CH and CH2 components. Cinnamic acids were increased, and proteinaceous materials and N-containing degradation or condensation compounds were absorbed or coprecipitated in RES. 2D HETCOR experiments indicated a close association between lignin and the residual carbohydrates. Ketones/aldehydes were not detected in the DAP, in contrast to a report in which an appreciable amount of ketones/aldehydes was generated from the acid pretreatment of a purified cellulose in the literature. This suggested that acid pretreatment may modify the structure of purified cellulose more than biomass and that biomass may be a better substrate than model biopolymers and compounds for assessing structural changes that occur with industrial processing. On the basis of NMR and wet chemical analyses, we found the following factors could cause the limitations to the effectiveness of enzymatic hydrolysis: (1) chemical modification of carbohydrates limited the biologically degradable carbohydrates available; (2) cinnamic acids in the residue accumulated; (3) accessibility was potentially limited due to the close association of carbohydrates with lignin; and (4) proteinaceous materials and N-containing degradation or condensation compounds were absorbed or coprecipitated.     

Kinetic study of the acid hydrolysis of various oligofructose samples

The kinetic of acid hydrolysis of five commercially available oligofructose samples used as food ingredients has been investigated as a function of the dry matter concentration, reaction pH, and temperature. The initial fructose release rate is found to be roughly proportional to the inverse of the average polymerization degree in number. A pseudo first order kinetic is found with respect to the fructosyl chain end concentration and to the proton concentration. Arrhenius plot is found to reasonably fit the data in a relatively wide temperature range (7-130 degrees C). The results allow the estimation of the fructose release rate in many foodstuff processing conditions.     

Determination of domoic acid in phytoplankton by high-performance liquid chromatography of the 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate derivative.

Domoic acid is the toxin that can cause amnesic shellfish poisoning. In this work, a new precolumn derivatization reagent, 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate, was used to react with domoic acid to form a stable unsymmetrical urea derivative, which was readily analyzed by reversed-phase HPLC with fluorescence detection. The one-step derivatizing method is rapid and straightforward. The method was applied to detect domoic acid in phytoplankton using a 3.9 x 150 mm Nova-Pak, 4 microm, C(18) column.     

Optimum conditions for formation of aflatoxin M1-trifluoroacetic acid derivative

Because thin-layer chromatographic (TLC) confirmation of identity and reverse-phase liquid chromatographic (LC) determination with fluorescence detection of aflatoxin M1 both require the derivative formed in the reaction of M1 and trifluoroacetic acid (TFA), various reaction conditions were studied to obtain complete derivative formation. Of the various organic solvents tested, the reaction between M1 and TFA proceeded best in the nonpolar solvents hexane and isooctane. Other parameters investigated were reaction temperature and time, aflatoxin M1 concentration, and solvent volume. The following procedure is considered optimum: 200 microL each of hexane and trifluoroacetic acid are mixed with M1 standard in a silylated glass vial or with milk residue in a regular glass vial with a Teflon-lined screw cap and heated 10 min at 40 degrees C. The mixture is evaporated to dryness under N2, and the derivative is saved for TLC or LC. No unreacted aflatoxin M1 was detected by reverse-phase LC after this procedure was incorporated for analysis of milk samples.     

Investigation of acid hydrolysis reactions of polyphosphates and phytic acid in Bray and Mehlich III extracting solutions

Soil test phosphorus (STP) (Bray P-1, Mehlich III) was developed to predict the plant available P in agricultural soils in the USA. The acidic STP extracting solutions were intended to extract phosphate from alkaline earth metal-based solubility products and or adsorbed species on amorphous metal hydroxides or phyllosilicates in soils. However, the accuracy of STP in predicting plant available phosphate has often been questioned due to potential acid hydrolysis of poly- and organic-P in soils during the STP extraction. The objective of the study was to investigate the acid hydrolysis reaction of reference poly- and organic-P compounds (1.25–3.75 mM) in Bray and Mehlich III extraction solutions using spectrophotometric methods. The results showed that both tripolyphosphate and pyrophosphate readily underwent the acid hydrolysis reaction over 2 h, whereas phytic acid did not hydrolyze. The results suggest an overestimation of dissolved reactive phosphate from these polyphosphates should be considered when evaluating the STP data.     

Quality protein maize: a biochemical study of enzymes involved in lysine metabolism.

Quality protein maize (QPM) varieties have been produced by the introduction of opaque-2 modifier genes. Two QPM varieties, BR451 and BR473, a wild type and an opaque-2 variety, have been used to study key enzymes controlling lysine metabolism in the endosperm during development. Aspartate kinase and homoserine dehydrogenase enzymes, which are involved in lysine and threonine biosynthesis, respectively, exhibited identical activity patterns during endosperm development, with a maximum specific activity at 16 days after pollination. The QPM varieties exhibited higher levels of aspartate kinase activity in the endosperm, suggesting an increased rate of lysine biosynthesis when compared to the opaque-2 and wild-type genotypes. Similar results were observed for the lysine ketoglutarate reductase and saccharopine dehydrogenase enzymes, which form a single bifunctional polypetide involved in endosperm lysine degradation. Both enzyme activities were strongly reduced in the opaque-2 maize variety when compared to the wild-type maize, whereas the QPM varieties exhibited even lower levels of lysine ketoglutarate reductase-saccharopine dehydrogenase activities when compared to the opaque-2 variety. The developmental pattern of enzyme activity showed a different profile when compared to the enzymes involved in lysine biosynthesis, with activity being detected only 12-16 days after pollination (DAP) and maximum activities approximately 24 DAP. These results also suggest that the modifier genes have intensified the effect of the opaque-2 mutation on lysine ketoglutarate reductase-saccharopine dehydrogenase. These alterations lead to an increase in soluble lysine in the endosperm of the QPM varieties when compared to the opaque-2 and wild type.     

Determination of gibberellic acid residues on fruits by synchronous scanning derivative spectrofluorometry

A synchronous derivative spectrofluorometric method is described for the determination of the plant growth regulator, gibberellic acid (GA3). The method is based on the formation of a fluorogen in concentrated sulfuric acid. The reaction is carried out at 85% sulfuric acid and in aqueous medium. The common fluorometric method with a linear dynamic range of 137-400 ppb, and a detection limit of 48 ppb is described. The synchronous first and second derivative method has linear dynamic ranges between 7.6-40 ppb and 12-40 ppb, with detection limits of 3.5 and 6.7 ppb, respectively. The influence of reaction variables and of other plant growth regulators present, and the application to residues on oranges, lemons, and grapes, are also described.     

NMR experiments of oleuropein biomimetic hydrolysis.

The oleuropein hydrolytic conversion, under biomimetic conditions, induced by the endogenous enzymatic system of the olive fruit, has been monitored by the intermediate derivative formation through (1)H and (13)C NMR spectra; the assigned molecular structures reveal the identity of new bioactive biophenolic metabolites, the hemiacetal aglycon, and the two epimeric dialdehydes, which influence the pathogen antagonism of olive fruits and the hedonistic-sensorial characteristics of olive oil.     

Reactivity of bovine whey proteins, peptides, and amino acids toward triplet riboflavin as studied by laser flash photolysis

The reaction between the triplet excited state of riboflavin and amino acids, peptides, and bovine whey proteins was investigated in aqueous solution in the pH range from 4 to 9 at 24 degrees C using nanosecond laser flash photolysis. Only tyrosine and tryptophan (and their peptides) were found to compete with oxygen in quenching the triplet state of riboflavin in aqueous solution, with second-order rate constants close to the diffusion limit, 1.75 x 10(9) and 1.40 x 10(9) L mol(-1) s(-1) for tyrosine and tryptophan, respectively, with beta-lactoglobulin and bovine serum albumin having comparable rate constants of 3.62 x 10(8) and 2.25 x 10(8) L mol(-1) s(-1), respectively. Tyrosine, tryptophan, and their peptides react with the photoexcited triplet state of riboflavin by electron transfer from the tyrosine and tryptophan moieties followed by a fast protonation of the resulting riboflavin anion rather than by direct H-atom abstraction, which could be monitored by time-resolved transient absorption spectroscopy as a decay of triplet riboflavin followed by a rise in riboflavin anion radical absorption. For cysteine- and thiol-containing peptides, second-order rate constants depend strongly on pH, for cysteine corresponding to pKaRSH = 8.35. H-atom abstraction seems to operate at low pH, which with rising pH gradually is replaced by electron transfer from the thiol anion. From the pH dependence of the second-order rate constant, the respective values for the H-atom abstraction (k = 1.64 x 10(6) L mol(-1) s(-1)) and for the electron transfer (k = 1.20 x 10(9) L mol(-1) s(-1)) were determined.     

Quantitative analysis of cystine, methionine, lysine, and nine other amino acids by a single oxidation--4 hour hydrolysis method

The sulfur-containing amino acids cystine and methionine play important roles in animal, especially avian, nutrition. Because these sulfur-containing amino acids are destroyed to varying extents by 6N HCl hydrolysis, oxidation and hydrolysis of cystine to cysteic acid and methionine to methionine sulfone have been widely used for determination of cystine and methionine. Lysine is considered the next limiting amino acid after the sulfur amino acids in poultry nutrition; therefore, determination of the amino acid content of rations focuses first on these 3 amino acids. The objective of this investigation was to establish whether lysine and other amino acids could be accurately determined in proteinaceous materials which had undergone performic acid oxidation. To perform this evaluation, lysine was determined in a variety of protein-containing materials both with and without performic acid oxidation. Performic acid oxidation followed by 6N HCl hydrolysis at 145 degrees C for 4 h allows accurate measurement of 3 amino acids especially important to poultry nutrition, cystine, methionine, and lysine, in a single preoxidized hydrolysate; this method can be extended to another 9 protein amino acids.     

Examination of thermal and acid hydrolysis procedures in characterization of soil organic matter

Abstract

Differential thermogravimetry (DTG), differential scanning calorimetry (DSC), and stepwise thermogravimetry (STG), together with two acid hydrolysis methods (hydrolysis with hydrochloric acid in a single step, and hydrolysis with sulfuric acid in two steps), were evaluated to determine the quality of four plant materials (Medicago sativa, Eucalyptus globulus, Quercus ilex, and Pinus halepensis) before and after mixing with a red earth. These quality indices were then compared with the same materials in the field, whether their decomposition could be predicted. All the thermal methods gave poor results. In both DTG and DSC, the presence of the mineral matrix gave rise to strong distortions in the spectra. Since the spectrum of any mixture is not simply the sum of the spectra of the two components (organic matter + mineral matter), these distortions could not be corrected by simply subtracting the spectrum of the red earth alone. STG trials also gave poor results, because the presence of the mineral matrix greatly increased the quality indices, and reduced the ability of the method to distinguish between organic matter qualities. In view of our results, the usefulness of thermal methods in the characterization of soil organic matter would seem to be restricted to certain organic horizons (L, F, and perhaps H). In contrast, methods based on acid hydrolysis were comparatively more satisfactory. Their resolution (ability to distinguish organic matter qualities) was much higher than that of thermal methods. However, they were able to distinguish carbon more accurately than nitrogen. The sulfuric acid method, unlike the hydrochloric acid method, was affected by the presence of a mineral matrix. While both methods could be improved, in their present form they seem to operate as good predictors of carbon and nitrogen mineralization.