Structural Investigation of the Antibiotic Sporaviridin. XV. Preparative-Scale Preparation of Sporaviridin Components by HSCCC

Abstract

Preparative-scale separation of the six sporaviridin components, very complicated basic glycoside antibiotics produced by Streptosporangium viridogriseum, was carried out by high speed countercurrent chromatography (HSCCC). The following two points, selection of proper solvent system and stationary phase retention, were carefully examined. Many two-phase solvent systems were searched, so that a solvent system, n-butanol:diethylether:water =10:4:12, was selected and other operating conditions were also optimized. The HSCCC was performed under the optimized conditions and gave satisfactory results.     

An efficient high‐speed countercurrent chromatography method for preparative separation of javanicin from Fusarium solani,a fungus isolated from the fruiting body of the mushroom Trametes trogii

To develop an efficient method for large preparation of javanicin from Fusarium solani, a rapid and simple method by high‐speed countercurrent chromatography was established based on average polarity (P′ values) and partition coefficients (K values) of crude samples. A suitable solvent system for high‐speed countercurrent chromatography was selected from many possible biphasic solvent systems. HSCCC was successfully applied to separate and purify javanicin, the main bioactive component of solid cultures of the fungus F. solani isolated from the fruiting body of Trametes trogii, with petroleum ether–ethyl acetate–methanol–water (4:3:2:1, v/v) as solvent system. A total amount of 40.6 mg of javanicin was obtained from 100 mg crude sample. The purity of javanicin was 92.2% with a recovery of 95.1%, as determined by high‐performance liquid chromatrography. The molecular structure was identified primarily by NMR and MS methods. The results indicated that high‐speed countercurrent chromatography could be a powerful technology for separating naphthoquinones from the solid cultures of the fungus F. solani. It is also of significance that the separation of javanicin from natural source was carried out for the first time utilizing high‐speed countercurrent chromatography.     

Isolation and structure determination of a new flavone glycoside from seed residues of seabuckthorn (Hippophae rhamnoides L.)

In this study, a valid method was established for the isolation and purification of flavone glycosides from Hippophae rhamnoides L. seed residues using high-speed counter-current chromatography in one step, with a solvent system of ethyl acetate-methanol-n-butyl alcohol-water (9:1:0.5:9, v/v/v/v). A total of 28.8 mg compound I and 57.3 mg compound II were obtained from 200 mg of flavone H-glycosides rich extract, with purities of 98.3 and 96.4%, respectively. The structures of two compounds were identified by MS and NMR. 3-O-β-D-Sophorosylkaempferol-7-O-{3-O-[2(E)-2,6-dimethyl-6-hydroxyocta-2,7-dienoyl]}-α-L-rhamnoside is compound I and compound II named hippophanone is a new compound were identified by MS and NMR. The method was efficient and convenient, which could be used for the preparative separation of flavone glycosides from H. rhamnoides L. seed residues.     

Separation of polar antioxidants from Rhizoma Polygonatum Odorati by high-speed counter-current chromatography with a hydrophilic solvent system

Some highly polar compounds are quality control makers for medicinal herbs. However, investigation of them has been hampered because the existing fractionation steps are difficult and laborious to purify them. Similar situations happen to Rhizoma Polygonatum Odorati, a widely used food supplement and medicinal herb with strong antioxidant activity, and up to date, only ethyl acetate fraction of Rhizoma Polygonatum Odorati has been comprehensively investigated. Here, HSCCC using a hydrophilic solvent system composed of n-butanol–ethanol–2 M ammonium sulfate (1:1:2, v/v/v) was performed to isolate highly polar antioxidants in n–butanol fraction of Rhizoma Polygonatum Odorati, guided by DPPH–HPLC experiment. Afterward, Sephadex LH-20 column chromatography eluted by methanol was selected to eliminate ammonium sulfate and purify co-eluted compounds in HSCCC collected fractions. Finally, nine compounds, including four nucleosides, cytidine (1), uridine (4), guanosine (5), and adenosine (8); two nucleobases, guanine (3), and adenine (6); and three amino acids, tyrosine (2), phenylalanine (7), and tryptophan (9) with purities over 98% were achieved and identified by UV, MS, and ¹H NMR data. Notably, compounds 1–9 were first reported in genus Polygonatum. The results indicated that the proposed method was an efficient approach to isolate and purify highly polar compounds from complex extracts.     

Application of the monomer reactivity ratios to the kinetic‐model discrimination and the solvent‐effect determination for the styrene/acrylonitrile monomer system

The impact of reactivity ratios determined with the Nelder and Mead simplex method on the kinetic‐model discrimination and the solvent‐effect determination for the styrene/acrylonitrile monomer system was investigated. For the monomer system, the penultimate unit effect was inversely proportional to the polarity of the solvent: acetonitrile < N,N‐dimethylformamide < methyl ethyl ketone < toluene. Quantitatively, the penultimate unit effect could be correlated with an absolute value of the difference between the standard deviation of the reactivity ratios determined for the terminal and penultimate models. By application of the F test, the penultimate model was justified for copolymerization in toluene. The conclusion was less certain for polymerization in methyl ethyl ketone. With a scanning procedure based on the simplex method, it was found that an equivalent representation of the copolymer‐composition data could be achieved with multiple sets of penultimate‐model reactivity ratios. However, the relationship between the triad‐sequence distribution and copolymer composition depended on the reactivity‐ratio set chosen for the microstructure determination. The microstructure calculated with the penultimate‐model reactivity ratios determined with the simplex method from the initial guess (r₁₁ = r₁, r₂₁ = 1/r₂, r₂₂ = r₂, r₁₂ = 1/r₁) did not obey the general “bootstrap effect” rule. This observation still requires some theoretical interpretation. © 2000 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 38: 846–854, 2000     

Studies on the Cationic Copolymerization of α-Pinene and Styrene with Complex SbCl3/AlCl3 Catalyst Systems. I. Effects of the Polymerization Conditions on the Copolymerization Products

Abstract

The copolymerization products of α-pinene and styrene were prepared with the compound catalyst system SbCl₃/AlCl₃ by changing the Sb/Al ratio, feeding monomer ratio, solvent, and polymerization temperature. The compositions of the copolymerization products were quantitatively characterized by the method of the combination of micro-ozonization and thin-layer chromatography in terms of the contents of the homopolymers and the copolymers containing high or low mole fractions of α-pinene, the yields of pure copolymer, and the monomer unit ratios (F ₁) of copolymers. The results show that it was easier to obtain the higher yield (up to 80%) of the pure copolymer with the complex catalyst system than with AlCl₃ alone. The F ₁ values could be controlled between 30 and 56% under the following polymerization conditions: Sb/Al  1/2, α-pinene/styrene  70%, and the conversion of styrene  90%.     

Separation of chlorogenic acid and concentration of trace caffeic acid from natural products by pH‐zone‐refining countercurrent chromatography

Chlorogenic acid and caffeic acid were selected as test samples for separation by the pH‐zone‐refining countercurrent chromatography (CCC). The separation of these test samples was performed with a two‐phase solvent system composed of methyl‐tert‐butyl‐ether/acetonitrile/water at a volume ratio of 4:1:5 v/v/v where trifluoroacetic acid (TFA; 8 mM) was added to the organic stationary phase as a retainer and NH₄OH (10 mM) to the aqueous mobile phase as an eluter. Chlorogenic acid was successfully separated from Flaveria bidentis (L.) Kuntze (F. bidentis) and Lonicerae Flos by pH‐zone‐refining CCC, a slightly polar two‐phase solvent system composed of methyl‐tert‐butyl‐ether/acetonitrile/n‐butanol/water at a volume ratio of 4:1:1:5 v/v/v/v was selected where TFA (3 mM) was added to the organic stationary phase as a retainer and NH₄OH (3 mM) to the aqueous mobile phase as an eluter. A 16.2 mg amount of chlorogenic acid with the purity of 92% from 1.4 g of F. bidentis, and 134 mg of chlorogenic acid at the purity of 99% from 1.3 g of crude extract of Lonicerae Flos have been obtained. These results suggest that pH‐zone‐refining CCC is suitable for the isolation of the chlorogenic acid from the crude extracts of F. bidentis and Lonicerae Flos.     

Thermochemical Investigations of Tautomeric Equilibrium. Variation of the Calculated Equilibrium Constant with Binary Solvent Composition

Abstract

A conventional nonelectrolyte solution model which has led to successful predictive equations for solute solubility and infinite dilution chromatographic partition coefficients is extended to systems containing a tautomeric solute dissolved in a binary solvent mixture. The derived expression predicts that the tautomeric solute concentration in a binary solvent is a geometric average of the pure solvent ratios and permits calculation of solute-solvent association constants from variation of the stoichiometric tautomeric solute concentration ratios as a function of binary solvent composition. Experimental data for phenylazonaphthol dissolved in aqueous-ethanol and aqueous-acetone solvent mixtures is discussed in relation to the theoretical model.     

PHENYLATION OF PYRIDINE BY PHOTOLYSIS OF DIPHENYL SULFONE

Abstract

The photolysis of diphenyl sulfone in neat pyridine or pyridine diluted with the organic solvent, with a high-pressure mercury arc lamp, was studied. The presence of acetone in the reaction system remarkably promoted photochemical conversion of the sulfone. The isomer distribution ratios (β > α > γ) of phenylpyridines produced during the reaction were clearly different from those (α > β > γ) so far reported for a free-radical phenylation of pyridine. Benzenesulfinic acid produced in the reaction was separated from the product mixture and identified as its S-benzylthiuronium derivative.     

Polymerizable Tautomers. VII. Radical Copolymerization of Ethyl 3-oxo-4-Pentenoate and Ethyl 4-Methyl-3-oxo-4-Pentenoate with Methyl Methacrylate

Abstract

Ethyl 3-oxo-4-pentenoate (EAA) and ethyl 4-methyl-3-oxo-4-pentenoate (EMAA) exhibit the coexistence of the ketonic and enolic forms in most organic solvents. Radical copolymerizations of EAA and EMAA with methyl methacrylate (MMA) were carried out at 60 °C in various solvents, and monomer reactivity ratios were estimated. There are minor solvent effects on monomer reactivity ratios r_MMA in both EAA/MAA and EM A A/MM A systems. On the other hand, r_EAA and r_MMA values greatly change with the solvent: The values decrease with an increase in the ketonic fraction of the polymerizable tautomers (EAA and EMAA). Regression analysis of the monomer reactivity ratios with the solvatochromic parameters reveals that polarity of the solvent is the major factor governing the relative reactivity.     

Chiral ligand exchange countercurrent chromatography: Enantioseparation of amino acids

This work deals with the enantioseparation of α‐amino acids by chiral ligand exchange high‐speed countercurrent chromatography using N‐n‐dodecyl‐l ‐hydroxyproline as a chiral ligand and copper(II) as a transition metal ion. A biphasic solvent system composed of n‐hexane/n‐butanol/aqueous phase with different volume ratios was selected for each α‐amino acid. The enantioseparation conditions were optimized by enantioselective liquid–liquid extractions, in which the main influence factors, including type of chiral ligand, concentration of chiral ligand and transition metal ion, separation temperature, and pH of the aqueous phase, were investigated for racemic phenylalanine. Altogether, we tried to enantioseparate 15 racemic α‐amino acids by the analytical countercurrent chromatography, of which only five of them could be successfully enantioseparated. Different elution sequence for phenylalanine enantiomer was observed compared with traditional liquid chromatography and the proposed interactions between chiral ligand, transition metal ion (Cu²⁺), and enantiomer are discussed.     

Light-scattering studies on solutions of cellulose in the ammonia/ammonium thiocyanate solvent system. I. Classical light-scattering

This article reports the use of classical light scattering to study cellulose in the NH3/NH4SCN solvent system. Three solvent compositions were used, 27.01 73.0,25.5/ 74.5, and 24.51 75.5 weight ammonia/weight ammonium thiocyanate. The coefficient, (dn/dc)_υ, was determined by back calculating using the molecular weight determined by solution viscometry in the solvent system cupriethylenediamine and the classical light-scattering results. Second virial coefficients were found to be similar to those values measured for cellulose in the FeTNa and LiCl/DMAC solvent systems. The characteristic ratios were found to vary with solvent composition with the highest values being at a composition of 25.5/74.5 weight ammonia/ weight ammonium thiocyanate. Persistence lengths were also found to vary with solvent composition with the highest value being 264 × 10^?8 cm at solvent composition 25.5/74.5.     

Metabolomics analysis of human plasma metabolites reveals the age- and sex-specific associations

Abstract

Objectives: The objective of this study was the investigation of age- and sex-associations in a set of blood plasma metabolites in healthy male and female subjects.

Methods: A comparison study design with male and female subjects of various ages was used. Metabolic profiling was performed using electrospray ionization tandem mass spectrometry that yielded 186 metabolite concentrations for each study participant. The key age-related metabolites were identified using an integrative analysis of absolute concentrations, metabolite ratios and the differential correlation of pairwise metabolite concentrations. All of the age-associated metabolites were adjusted prior to the analysis to account for differences in Body Mass Index (BMI).

Results: A total of 236 plasma samples from 140 female and 96 male subjects aged 20 to 82 years-old were collected and analyzed in the study. 13 and 14 age-associated metabolites (|r|?>?0.33 and p < 6.6×10^?5), 438 and 337 age-associated metabolite ratios (|r|?>?0.37 and p < 3.5×10^?6), and 5 and 10 core metabolites were discovered in the female and male groups, respectively. 80% of the metabolites displaying associations with age belonged to sphingolipids and phosphatidylcholines, and the two sexes shared less than 50% of the age-associated metabolites.

Conclusion: The study found that changes in metabolite concentrations, metabolite ratios and differential correlations were age and sex-specific.     

Studies on Copolymerization of Acrylamide and N-Vinylpyrrolidone

Abstract

Copolymers of acrylamide and N-vinylpyrrolidone were prepared using different techniques, viz., solution, precipitation, and inverse emulsion. Xylene was used as a continuous medium and sorbitane monooleate as a surfactant for the inverse emulsion technique. Acetone was used as a solvent in the precipitation technique. The initiator used in all the reactions was 2,2-azobis(2-amidinopropane)dihydrochloride. C, H, and N elemental analysis was used to determine the compositions of the copolymers formed. Reactivity ratios as determined by the Fineman and Ross method were found to be 0.61 and 0.05 for acrylamide and n-vinylpyrrolidone, respectively. Intrinsic viscosity measurements were made at temperatures ranging from 16 to 75°C.     

Quantitative Determination of Digitalis Glycosides in Digitalis Purpurea Leaves by Reversed-Phase Thin-Layer Chromatography

Abstract

A densitometric reversed-phase thin-layer chromatographic(RP-TLC) method for the determination of digitalis glycosides in Digitalis purpurea leaves has been developed. The procedure involves extraction of dry leaf powder with ethanol/chloroform (2:1) and clean-up by Sep-Pak cartridges prior to RP-TLC analysis. RP-TLC was performed on an octadecylsilyl bonded silica gel plate, using a developing solvent of acetonitrile/0.5 M NaCl(1 : 1) for secondary glycosides and acetonitrile/0.5 M NaCl(2 : 3)for primary glycosides. The plate was scanned with a reflectance densitometer at 222 nm. Quantitation of these glycosides was carried out by the internal standard method. The amounts of digitoxin, gitoxin, gitaloxin, purpurea glycoside A, purpurea glycoside B, and glucogitaloxin in a leaf powder sample of Digitalis purpurea were estimated from the calibration graphs for pure glycosides.     

Ester and Ketone Groups Substituted Mono- and Di- Azo-coupled Azocalix[4]arenes as Extractant for Hg+ and Hg2+, or Cr3+Cations

This article describes extraction properties of mono- (A1–A8) and di- (B1–B8) substituted azocalix[4]arene analogues. The ionophore solvent extractions of alkaline-earth (Sr²⁺), basic metal (Pb²⁺) and transition metal cations (Ag⁺, Hg⁺, Hg²⁺, Co²⁺, Ni²⁺, Cu²⁺, Cr³⁺) from aqueous phase to organic phase were carried out by azocalix[4]arene derivatives. It has been observed that they show a good extraction behavior toward selected heavy metal (Hg) and toxic metal (Cr), while A4 and B4 prefer Hg⁺, Hg²⁺ and Cr³⁺ among transition metal cations, respectively. The azocalix[4]arenes (A1–A8) and (B1–B8) are not efficient extractants for all of the selected metal cations, whereas A4 and B4 are selective only for Hg metal cation.     

溶剂对钯催化的叔丁基乙炔低聚反应化学选择性的调控作用

报道了溶剂对钯催化的叔丁基乙炔低聚反应化学选择性的调控作用. 反应可在苯-正丁醇双组分溶剂体系中顺利进行, 当双组分溶剂体系中苯占优势比例时, 反应发生递次的三分子炔烃顺式插入, 经由顺式s-烯钯中间体生成环三聚产物1,3,5-三叔丁基苯; 而当双组分溶剂中正丁醇组分上升至一定比例, 反应选择性生成(3Z,5Z)-2,2,7,7-四甲基- 3,6-二氯-3,5-辛二烯或(3Z,5Z)-2,2,7,7-四甲基-3,6-二溴-3,5-辛二烯, 这是由于正丁醇可显著加快C—Pd σ键的断裂, 并与叔丁基乙炔、σ-烯钯中间体形成弱氢键作用力, 同时也与Pd(II)和Cu(II)等离子存在配位效应. 在强极性质子溶剂H₂O中, 反应生成偶联双炔: 2,2,7,7-四甲基-3,5-辛二炔. 文中就反应溶剂体系、钯铜催化剂及反应可能机理等分别进行了探讨.     

Three‐phase solvent systems for the comprehensive separation of a wide variety of compounds from Dicranostigma leptopodum by high‐speed counter‐current chromatography

A three‐phase solvent system was efficiently applied for high‐speed counter‐current chromatography to separate secondary metabolites with a wide range of hydrophobicity in Dicranostigma leptopodum. The three‐phase solvent system of n‐hexane/methyl tert‐butyl ether/acetonitrile/0.5% triethylamine (2:2:3:2, v/v/v/v) was selected for high‐speed counter‐current chromatography separation. The separation was initiated by filling the column with a mixture of intermediate phase and lower phase as a stationary phase followed by elution with upper phase to separate the hydrophobic compounds. Then the mobile phase was switched to the intermediate phase to elute the moderately hydrophobic compounds, and finally the polar compounds still retained in the column were fractionated by eluting the column with the lower phase. In this research, 12 peaks were eluted out in one‐step operation within 110 min, among them, eight compounds with acceptable purity were obtained and identified. The purities of β‐sitosterol, protopine, allocryptopine, isocorydione, isocorydine, coptisine, berberrubine, and berberine were 94.7, 96.5, 97.9, 86.6, 98.9, 97.6, 95.7, and 92.8%, respectively.     

Development of a general separation strategy by countercurrent chromatography using sanshools from Zanthoxylum bungeanum oleoresin as a case study

Three kinds of sanshools were separated from Zanthoxylum bungeanum oleoresin by high-speed countercurrent chromatography. Sanshools are a series of amide compounds extracted from the Zanthoxylum bungeanum. Due to similar structures, polarities, and dissociation constants, it was challenging to select an appropriate solvent system for their complete separation by countercurrent chromatography. To address this challenge, a solvent-system-selection strategy was proposed to identify a relatively suitable solvent system. Additionally, a separation procedure incorporating multi-elution modes selection was established to separate similar compounds in a logical order. Ultimately, a solvent system comprising n-hexane:ethyl acetate:methanol:water in a ratio of 19:1:1:5.67 was selected. Three amide compounds with high purity were obtained through the use of recycling elution mode to improve separation resolution: hydroxy-ε-sanshool (8.4 mg; purity: 90.64%), hydroxy-α-sanshool (326.4 mg; purity: 98.96%), and hydroxy-β-sanshool (71.8 mg; purity: 98.26%) were obtained from 600 mg sanshool crude extract. The summarized solvent-system-selection strategy and separation procedure incorporating multi-elution modes may instruct countercurrent chromatography users, particularly novices, seeking to separate compounds with highly similar chemical properties.     

Structural parameters of concentrated aqueous solutions of LiCl under extreme conditions,as calculated by the method of integral equations. Effect of temperature

Peculiarities of structure formation of aqueous LiCl solutions at different salt : water molar ratios (LiCl : n H₂O, n = 3.15, 8.05, 14.90) under conditions of isobaric heating (p = 100 bar, T = 298, 323—523 K, T = 50 K) were studied by the method of integral equations. Heating of LiCl : 14.90H₂O solution was found to lead to disappearance of tetrahedral ordering of solvent molecules, appreciable weakening of the coordination abilities of both ions, and to an increase of the number of contact ion pairs and a decrease of the number of solvent-separated ion pairs. For the LiCl : 8.05H₂O system, the tetrahedral structure of the solvent disappears at a lower temperature and heating has a less pronounced effect on the coordination and associative abilities of the ions. In the LiCl : 3.15H₂O solution, tetrahedral ordering of the solvent molecules disappears at 298 K and the number of contact ion pairs decreases as temperature increases. Other structural changes in this system upon heating are similar to those found for the LiCl : 14.90H₂O and LiCl : 8.05H₂O solutions.