One-pot preparation of quercetin using natural deep eutectic solvents

In this study, we have established a green and efficient preparation method of quercetin. Rutin was first extracted from Sophora japonica using natural deep eutectic solvents (NADESs), then hydrolyzed into quercetin by rutin degrading enzyme (RDE) obtained from germinated tartary buckwheat in situ. Rutin solubility tests showed that most of the 11 NADESs increased the solubility of rutin by 67-3116 times compared to water. Thus, NADESs could be prior candidate to extract rutin. Extraction efficiency of rutin varied with different NADESs, and a maximum of 291.57 mg g⁻¹ was achieved in NADES ChGly, which was prepared by mixing choline chloride and glycerol at a molar ratio of 1:1. After that hydrolysis was performed directly in extraction system by adding RDE with degradation rate of up to 8.36 mg min^-1·L^-1. Our findings suggest that preparation of quercetin using NADESs was simple and feasible to operate, environmentally friendly, efficient, and inspired the preparation method of bioactive components from a new perspective.     

Rapid solubility determination of the triterpenes oleanolic acid and ursolic acid by UV-spectroscopy in different solvents

Salvia triloba (Greek sage) has been used for the treatment of various diseases and contains two bioactive triterpene acids of major interest: oleanolic acid (OA) and ursolic acid (UA). The determination of the solubility of OA and UA in different solvents is a prerequisite to select the optimal solvent. The main goal of this work was to develop a quick method of predicting the solubility of OA/UA in different solvents to get a first indication of which solvents could be considered suitable for extraction from any plant material containing at least one of these triterpenes. A novel and simple ultra-violet spectroscopy method was developed for this purpose.The best solubilities were determined in THF, dioxane and n-butanol as well as in blends of dioxane and n-butanol.     

1-Aminocyclopentane-1,2,4-tricarboxylic acids screening on glutamatergic and serotonergic systems

Enantiopure constrained 1-aminocyclopentane-1,2,4-tricarboxylic acids containing the glutamic acid skeleton were prepared as two diastereomers characterized by having the carboxylic groups in position two and four cis-oriented to each other and trans with respect to 1-carboxylic group and all cis-oriented carboxylic groups, respectively. A biochemical screening of activity of the above amino acids was investigated on glutamate and 5-HT receptors to find a possible metabotropic agonist, acting on the serotoninergic system.     

Effect of the cation and the anion of an electrolyte on the solubility of DL-aminobutyric acid in aqueous solutions: measurement and modelling

The solubilities at 298.2 K of dl-aminobutyric acid in aqueous solutions of NaCl, KCl, NaNO(3) and KNO(3) were measured. The solubility of DL-aminobutyric acid was found to be influenced by the concentration and by the nature of both the cation and the anion of the electrolyte. Comparison of the results obtained in this study and those for other amino acids reported in the literature, indicates that the structure of the hydrocarbon backbone of an amino acid plays an important role in the interactions of an amino acid with an electrolyte. A thermodynamic model has been used to correlate the solubilities of DL-aminobutyric acid in aqueous electrolyte solutions. The activity coefficients of the amino acid in the electrolyte solutions, were represented by a model proposed by Khoshkbarchi and Vera [M.K. Khoshkbarchi, J.H. Vera, AIChE J. 42 (1996) 2354; M.K. Khoshkbarchi, J.H. Vera, Ind. Eng. Chem. Res. 35 (1996) 4755]. This model, which considers a combination of both long- and short-range interactions, contains only two adjustable parameters. All other parameters are available in the literature. The model can accurately correlate the solubility of dl-aminobutyric acid in aqueous solutions of electrolytes.     

Natural deep eutectic solvents are viable solvents for plant cell culture-assisted stereoselective biocatalysis

We report a plant (Beta vulgaris L. subsp. vulgaris.) cell culture-mediated reduction of prochiral 1-(3,4-dimethylphenyl)ethanone into the chiral alcohol (1R)-1-(3,4-dimethylphenyl)ethanol in natural deep eutectic solvents (NADES). To the best of our knowledge, this is the first report on plant culture cell behaviour during incubation in these solvents, where the novelty is transformation in callus culture. Three different choline chloride-based NADES (aqueous solutions containing 30, 50 and 80 % water [w/w]) containing sugar (glucose) or polyalcohols (glycerol and ethylene glycol) were screened for conversion and enantiomeric excess during the bioreduction. Both the conversion and the enantiomeric excess differed considerably when using different hydrogen bond donors, with the (R)-alcohol configuration predominating in the reactions conducted in most NADES. Changing the water content in NADES also altered the enantioselectivity. Testing the biocompatibility of NADES with sugar beet cell cultures revealed that NADES cause permeabilisation of cell membranes, leading to stress conditions that change plant metabolism. The potential for recycling and reusing plant biomass was tested for sugar beet cell cultures. The results indicate that recycling may be possible after 3–7 days of incubation, but longer incubations lead to too high a toxicity to cellular metabolism.     

Olfactory sensitivity and odor structure-activity relationships for aliphatic carboxylic acids in CD-1 mice

Using a conditioning paradigm, the olfactory sensitivity of CD-1 mice for a homologous series of aliphatic n-carboxylic acids (ethanoic acid to n-octanoic acid) and several of their isomeric forms was investigated. With all 14 odorants, the animals significantly discriminated concentrations as low as 0.03 ppm (parts per million) from the solvent, and with four odorants the best-scoring animals even detected concentrations as low as 3 ppt (parts per trillion). Analysis of odor structure-activity relationships showed that the correlation between olfactory detection thresholds of the mice for the unbranched carboxylic acids and carbon chain length can best be described as a U-shaped function with the lowest threshold values at n-butanoic acid. A significant positive correlation between olfactory detection thresholds and carbon chain length of the carboxylic acids with their branching next to the functional carboxyl group was found. In contrast, no such correlation was found for carboxylic acids with their branching at the distal end of the carbon chain relative to the functional carboxyl group. Finally, a significant correlation was found between olfactory detection thresholds and the position of the branching of the carboxylic acids. Across-species comparisons suggest that mice are more sensitive for short-chained (C(2) to C(4)) aliphatic n-carboxylic acids than other mammalian species, but not for longer-chained ones (C(5) to C(8)). Further comparisons suggest that odor structure-activity relationships are both substance class- and species-specific.     

Lipase catalyzed esterification of glycidol in organic solvents

We studied the resolution of racemic glycidol through esterification with butyric acid catalyzed by porcine pancreatic lipase in organic media. A screening of seven solvents (log P values between 0.49 and 3.0, P being the n-octanol-water partition coefficient of the solvent) showed that neither log P nor the logarithm of the molar solubility of water in the solvent provides good correlations between enantioselectivity and the properties of the organic media. Chloroform was one of the best solvents as regards the enantiomeric purity (e. p.) of the ester produced. In this solvent, the optimum temperature for the reaction was determined to be 35 degrees C. The enzyme exhibited maximum activity at a water content of 13 +/- 2% (w/w). The enantiomeric purity obtained was 83 +/- 2% of (S)-glycidyl butyrate and did not depend on the alcohol concentration or the enzyme water content for values of these parameters up to 200 mM and 25% (w/w), respectively. The reaction was found to follow a BiBi mechanism. (c) 1993 John Wiley & Sons, Inc.     

Cadmium mobilisation and plant availability – the impact of organic acids commonly exuded from roots

The present work highlights metal-organic acid interactions with special reference to their plant availability. Pot experiments were conducted to investigate the effect of various organic (carboxylic and amino) acids on the uptake and translocation of root-absorbed Cd by maize (Zea mays) plants grown in sand and soil culture. Statistically significant increases in Cd accumulation from Cd-treated plants in the presence of increasing concentrations of organic acids, suggest the existence of Cd-organic acid interactions in the soil-plant system. In order to support the above hypothesis of formation of organically bound Cd, separate experiments were performed to synthesize and estimate its various forms viz. cationic, anionic and neutral. The chemical nature of the organically bound forms was ascertained by electrophoretic experiments. Amino acids have been found to be less effective in the mobilisation of cadmium compared to carboxylic acids. The results are discussed on the basis of the potential of organic acids to form complexes with Cd.     

Deterrent effect of carboxylic acids on cabbage root fly oviposition

In laboratory experiments, 11 selected carboxylic acids were tested to determine which part of the sinapic acid molecule is responsible for deterring cabbage root fly from laying its eggs on otherwise-acceptable cauliflower host-plants. The deterrent effect was only obtained with compounds containing at least one carboxylic group in the molecule. Hence, the aliphatic acids were as deterrent as the aromatic acids to the fly and all the carboxylic acids were as deterrent as sinapic acid, reducing oviposition by > 50%. The inclusion of two carboxylic groups in the molecule, (e.g. phthalic acid and oxalic acid) did not increase the deterrent effect observed with sinapic acid. Some of the long chain fatty acids, with low volatility, low water solubility and thus greater persistence, offer practical opportunities for deterring Delia radicurn from laying its eggs on plants in the field.     

基于天然低共熔溶剂的甜叶菊中甜菊糖绿色提取方法及优化

尝试利用天然低共熔溶剂(NADES)提取甜叶菊(Stevia rebaudiana)中的甜菊糖, 探索一种高效、绿色和环保的甜菊糖提取新方法。以甜叶菊干叶为原料, 对照传统提取溶剂水, 以甜菊糖中甜菊苷和莱鲍迪苷A的提取浓度作为指标, 筛选出最优的NADES提取配方, 然后通过Box-Behnken响应面法对NADES提取甜叶菊中甜菊糖的工艺条件进行筛选优化。结果表明, 提取效率最高的NADES配方为1,2-丙二醇:甘油:水=8:1:1 (v/v/v), 提取的甜菊苷浓度为2.59 mg∙mL^-1, 比水提取高16.40%, 提取的莱鲍迪苷A浓度为1.06 mg∙mL^-1, 比水提取高12.62%; 通过响应面法得到最优提取条件: 提取时间90分钟, 提取温度60°C, 超声功率为80 J∙s^-1, 预测甜菊苷提取浓度为3.49 mg∙mL^-1, 莱鲍迪苷A提取浓度为1.43 mg∙mL^-1, 与实验验证值(甜菊苷浓度为3.48 mg∙mL^-1, 莱鲍迪苷A浓度为1.42 mg∙mL^-1)接近。在最优条件下, 甜菊苷提取浓度比初始条件提高了34.36%, 莱鲍迪甘A提取浓度比初始条件提高了33.96%。NADES绿色环保, 且提取效率高于传统溶剂, 可用于甜叶菊中甜菊糖的绿色提取; 同时, 该提取方法可为后续推广至其它大宗经济植物类天然产物的绿色工业生产提供参考。     

Recognition modes of hypoxanthine,xanthine and their derivatives by amino acid carboxylic group: UV spectroscopic and quantum chemical data

UV absorption spectra of Hyp, Xan, their nucleosides and methyl derivatives were studied in anhydrous dimethylsuloxide and the changes in these spectra on the interactions with neutral and deprotonated carboxylic groups of amino acids were traced. By the semiempirical quantum-chemical method MNDO/H it was shown, that interaction with carboxylate-ion fixes Hyp as the rare N7H enolic tautomer and converts Xan into its N9H diketo tautomeric form with a probable admixture of the N7H O6-enolic form. Significant changes in the UV spectra of Xan, m3Xan, m9Xan and X under interaction with carboxylate-ion are determined by essential contribution to complex formation of proton transfer from bases to ligands, m9Xan and X proving to be slightly protonated even by the solvent. The methylation of the N7 position in m7I and m7X was established to result in the practical absence of their interactions with carboxylate-ion and initiation of a new ability of forming complex with the neutral carboxylic group. Substitution of the C8H group by N in 8-azaXan does not change the interaction specificity of the base with two forms of carboxylic group.     

Toxicity of organic solvents used in situ in microbial fermentation

Summary Toxicity of organic solvents for in situ solvent extraction of carboxylic acids from aqueous solutions was studied on bacteria Bacillus subtillis and Acetobacter spp., yeasts Kluyveromyces marxianus and Pichia fermentans and fungi Aspergillus terreus and Rhizopus arrhizus. It has been found that trialkylamine and tributylphosphate are much more toxic than other carriers as trialkylphosphineoxide, triisobutylphosphinesulfide and trihexylphosphate. For R. arrhizus trihexylphosphate was found to be the less toxic solvent.     

Solvation properties of ubiquinone-10 in solvents of different polarity

The solvation properties of ubiquinone-10 and ubiquinol-10 in a wide variety of solvents of polarity varying from alkanes to water are reported. Greatest solubility is observed in solvents of intermediate polarity and particularly where low polarity is combined with a pronounced tendency to interact with the benzoquinone substituent of the ubiquinone molecule. This includes solvents like chloroform and benzene. Ubiquinone-10 is somewhat less polar than ubiquinol-10 as judged by comparative solubilities of the two molecules. Proton-NMR chemical shift measurements and aggregation studies in selected solvents indicate that in ubiquinone-10 in the liquid phase and in solution in hydrocarbons like dodecane the molecules have a preferred association possibly involving stacking of the benzoquinone rings. Surface balance studies indicated that the surface-active character of ubiquinone-10 is relatively weak and only in a comparatively polar and highly structured solvent, formamide, was there evidence of an effect on surface tension of the solvent. The critical micelle concentratiom in this solvent was estimated to be about 5 M on the basis of surface tension measurements. Ubiquinone-10 is well known to form virtually insoluble monolayers at the air/water interface. Studies of the partition of ubiquinone-10 in binary mixtures of solvents suggest that the interaction of the benzoquinone ring substituent with structured polar solvents is considerably weaker than the internal cohesion between molecules of the solvent. No evidence on the basis of wide-angle X-ray diffraction measurements was obtained to indicate that solvent molecules were a component of the crystal lattice of ubiquinone-10 that had precipitated from solvent mixtures.     

Selector Screening for Enantioseparation of dl‐α‐Methyl Phenylglycine Amide by Liquid–Liquid Extraction

Enantioseparation through liquid extraction technology is an emerging field, e.g., enantioseparations of amino acids (and derivatives thereof), amino alcohols, amines, and carboxylic acids have been reported. Often, when a new selector is developed, the versatility of substrate scope is investigated. From an industrial point of view, the problem is typically approached the other way around, and for a target racemate, a selector needs to be found in order to accomplish the desired enantioseparation. This study presents such a screening approach for the separation of the enantiomers of dl ‐α‐methyl phenylglycine amide (dl ‐α‐MPGA), a model amide racemate with high industrial relevance. Chiral selectors that were reported for other classes of racemates were investigated, i.e., several macrocyclic selectors and Pd‐BINAP complexes. It appeared very challenging to obtain both high extraction yields and good enantioselectivity for most selectors, but Pd‐BINAP‐based selectors performed well, with enantioselectivities up to 7.4 with an extraction yield of the desired enantiomer of 95.8%. These high enantioselectivities were obtained using dichloromethane as solvent. Using less volatile chlorobenzene or 1‐chloropentane, reasonable selectivities of up to 1.7 were measured, making these the best alternative solvents for dichloromethane. Chirality 27:123–130, 2015. © 2014 Wiley Periodicals, Inc.     

Antimicrobial activity of basic cholane derivatives. X. Synthesis of 3 alpha- and 3 beta-amino-5 beta-cholan-24-oic acids.

A simple and convenient route to 3 alpha- and 3 beta-amino-5 beta-cholan-24-oic acids was developed via Leuckart-Wallach amination reduction and subsequent acid hydrolysis. Two epimeric formylamino derivatives were produced (alpha and beta), approximately in a 1:1 ratio, as determined by 13C nuclear magnetic resonance spectroscopy. The two isomers were separated by making use of their different solubilities in ethyl ether. The absolute configuration of the two amino acids was assigned by comparison with authentic reference samples.     

Determination of isokinetic ratios necessary for equimolar incorporation of carboxylic acids in the solid-phase synthesis of mixture-based combinatorial libraries

The methods used to study the relative reaction rates of 45 different aliphatic and aromatic carboxylic acids when coupled to resin-bound amino acid amides is described. Competition experiments involving the coupling of incoming carboxylic acids to resin-bound amino acid amides were performed. The relative composition of each N-acylated amino acid amide in the resulting mixtures was compared to controls prepared by physically mixing equal aliquots of individual compounds in order to study the relative reaction rates of the incoming carboxylic acids. The ratios of the incoming carboxylic acids were then iteratively adjusted to yield as close to equimolar products as possible. As expected, the steric and electronic nature of the incoming carboxylic acids was found to influence their relative reaction rates. The steric hindrance of the resin-bound amino acid appears to have a proportional effect on the reaction rates of the incoming carboxylic acids. N-acylated amino acid amides in the final mixtures, prepared using the final isokinetic ratios, were found to be approximately equimolar.     

Natural deep eutectic systems as alternative nontoxic cryoprotective agents

Natural deep eutectic systems (NADES) are mostly composed of natural primary metabolites such as sugars, sugar alcohols, organic acids, amino acids and amines. These simple molecules have been identified in animals living in environments with extreme temperature amplitudes, being responsible for their survival at negative temperatures during winter. Herein, we report for the first time the use of NADES based on trehalose (Treh) and glycerol (Gly) in cryopreservation, as cryoprotective agents (CPA). The evaluation of the thermal behaviour of these eutectic systems, showed that NADES have a strong effect on the water crystallization/freezing and melting process, being able to reduce the number of ice crystals and hence ice crystal damage in cells, which is a crucial parameter for their survival, upon freezing. Using this NADES as CPA, it is possible to achieve similar or even better cellular performance when compared with the gold standard for cryopreservation dimethyl sulfoxide (DMSO). In this sense, this work relates the physical properties of the NADES with their biological performance in cryopreservation. Our comprehensive strategy results in the demonstration of NADES as a promising nontoxic green alternative to the conventional CPA's used in cryopreservation methods.     

Structural requirements for nucleophilic substrates of carboxypeptidase Y

The composition and structural aspects of the amino and carboxylic acid groups required for incorporation into peptides by transpeptidation and inhibition of hydrolysis in carboxypeptidase Y-catalyzed reactions were studied. Separation of these two groups by even one carbon prevents incorporation by transpeptidation and does not inhibit incorporation of other amino acids into model peptides. Substitution of phosphonic or sulfonic acids for the carboxylic acid group also results in loss of incorporation by transpeptidation. Only the sulfonic acid analog of glycine causes inhibition of hydrolysis and this inhibition is lost when serine is included in the reaction. d-Serine is not incorporated by carboxypeptidase Y, and its presence in the reaction mixture does not inhibit the incorporation of the L-isomer.     

Interaction of purple membrane with solvents. II. Mode of interaction.

Using the solubility parameter mapping technique (Eisenbach, M., Caplan, S.R. and Tanny, G (1979) Biochim. Biophys. Acta 554, 269-280) we studied spectroscopically the mode of interaction between the purple membrane of Halobacterium halobium and pure organic solvents or solvent mixtures. Although the interacting solvents formed a well-defined closed region in the interaction maps, mapping the modes of interaction did not reveal a closed region for each spectrally classifiable type. A suggested interpretation for this is that interaction with the purple membrane chromophore requires that a solvent (or solvent mixture) possess apolar groups in order to obtain access to the chromophore, together with a polar character and hydrogen-bonding capacity. The mode of interaction, however, is dependent on the specificity of the reactive group of the solvent for retinal, and this has nothing to do with membrane properties. We also examined the influence of the duration of the interaction and of illumination. Some solvents appeared to react more sluggishly than others, but no generalization in terms of the solubility parameter mapping was found, probably because the map describes thermodynamic rather than kinetic phenomena. The only effect of illumination was to enhance the reaction of some of these solvents. It did not change the solubility parameters of purple membrane.     

Effect of cosolvents on solubility of protein in equilibrium between different states

This paper describes a theoretical work on the solubility of a protein in equilibrium between different forms such as isomerization (A ? B) and dimerization (2A ? B). Under the assumptions that A and B have different solubilities and form their own solid phases (in the form of either precipitation or crystal), it was shownthat only either A or B precipitates at a given condition, another form being in equilibrium in solution with the precipitating form, provided that the solubility ratio of A to B is not identical to their equilibrium composition in the solution phase determined by the equilibrium constant. It follows, then, that the total concentration of the protein in the solution phase can be calculated by summing the solubility of the precipitating form and the concentration in solution of another form in equilibrium with the precipitating form. Assuming various preferential protein-solvent interactions for the two forms in solution as well as solid phases, dependences of the total protein solubility on the additive concentration were examined. It was shown that this dependence may be complex, showing maximum, minimum, or inflection point, depending on the preferential protein interactions with solvent components.