Acetonitrile Dichloromethane Research Articles

The Effect of Weakly Coordinating Solvent and Counter Anion on Diphosphine‐Palladium(II) Catalytic Reactivity for Copolymerization of 1‐Butene with Carbon Monoxide in Aqueous Media

AbstractWater soluble bis(acetonitrile) diphospalladium (II) complexes cis‐[Pd(L1–L5)(NCCH3)2][BF4]2 (C1–C5) were obtained by the reaction [Pd(NCCH3)4][BF4]2 with diphosphines ligands L1–L5 (CH2(CH2PR2)2: where R=(CH2)nP(O)(OEt)2, n=2–4, n=6 and n=8)) in dichloromethane–acetonitrile. The isolated complexes were characterized by elemental analysis, FTIR, 1H‐ and 31P‐NMR spectroscopy. The complexes were applied as catalysts for the copolymerization of 1‐butene with carbon monoxide in aqueous media. The present work emphasized on the effect of weakly coordinated solvent and counterion on the catalytic behavior of cis‐[Pd(L1–L5)(NCCH3)2][BF4]2 complexes compared to previously prepared cis‐[ Pd(L1–L5)(OAc)2] (A1–A5). The exchange of π‐bonding NCCH3 ligand with π‐bonding CO and alkene was more favored than the exchange of σ‐bonding OAc− as reflected by the much higher catalytic activity (TON) of the former compared to the latter. In order to compare the catalytic efficiency of water soluble complexes with the performance of a standard, the water insoluble complex, cis‐[Pd(CH2(CH2P(CH2CH3)2)2(NCCH3)2][BF4]2, C6, was prepared and incorporated in the investigations. The isolated alternating 1‐butene/CO copolymer exhibits glass transition temperature of Tg=−3–−15 °C, which is in the range desirable for latex emulsion application. The solid content is up to 21 % with high molecular weight (Mw=4×104 g mol−1) and narrow polydispersities (Mw/Mn=2–3). The C5 complex was found to be fourteen times more active in aqueous polymerization medium (6369 mol substr. mol Pd−1 h−1) than C6 complex in nonaqueous medium (441 mol substr. mol Pd−1 h−1).

Sequential combination of solid-phase sorbents to enhance the selectivity of organosulfur compounds for flavour analysis.

Flavour analysis remains challenging due to the range of selectivity demands, from the extraction of multiclass volatile compounds to the purification of low-concentration odourants (e.g. organosulfur compounds) amidst the high food matrix noise. In this study, the varying selectivities of solid-phase extraction (SPE) were leveraged upon for both multiclass and organosulfur compound analysis, using coffee as a model matrix. Polymeric SPE (Bond Elut ENV) was screened for significant (p<0.05) parameters affecting the recovery of 37 multiclass compounds, and the most influential parameters were optimised using a Box-Behnken design (elution solvent of 67:33 dichloromethane:ethyl acetate, loading pH of 4.8, and wash solvent of water). Following this, low-concentration organosulfur compounds which were challenging to detect in the complex coffee matrix were purified by adding a sequential SPE step for selectivity. A silver-based ligand-exchange SPE step (MetaSEP IC-Ag) was optimised for organosulfur compound recovery (wash solvents of dichloromethane and 43% acetonitrile in dichloromethane, elution solvent of 90mmol/L 1,4-dithiothreitol in dichloromethane). This was found to be complementary to polymeric SPE's matrix clean-up effect (which improved average organosulfur compound recovery by 5.45 times). Finally, both multiclass and sequential organosulfur extraction techniques demonstrated good performance in terms of reproducibility (1.0-9.0%) and linearity (R2>0.995), allowing for the detection of 3-mercapto-3-methylbutyl formate (3.741±0.387ng/mL) in coffee samples. In conclusion, this study highlights the potential for SPE to address a variety of complex flavour analysis demands with the appropriate selection and combination of solid-phases.

Isolation of Four Major Compounds of γ-Oryzanol from Rice Bran Oil by Ionic Liquids Modified High-Speed Countercurrent Chromatography and Antimicrobial Activity and Neuroprotective Effect of Cycloartenyl Ferulate In Vitro

This study presents an effective high-speed countercurrent chromatography method of two-phase solvent system combined with ionic liquids for the isolation and purification of four major compounds of γ-oryzanol from rice bran oil. The conditions of high-speed countercurrent chromatography, including solvent systems and the content of ionic liquids, were optimized to improve separation efficiency. The optimal solvent system was n-hexane–dichloromethane–acetonitrile–[C4mim][PF6] at a volume ratio of (10:1:10:0.15, v/v). Adding a small amount of ionic liquids significantly improved the separation efficiency. Four compounds of γ-oryzanol, cycloartenyl ferulate, 24-methylene cycloartanyl ferulate, campesteryl ferulate, and sitosteryl ferulate were successfully purified in one run from rice bran oil with the purities of 95.25%, 92.48%, 92.47%, and 93.43%, respectively. The result of bioassay showed that rice bran oil and separated individual cycloartenyl ferulate could significantly inhibit mycelium growth of crop diseases Rhizoctonia solani and Fusarium graminerum, the bioactivity of cycloartenyl was higher than that of the rice bran oil. The cellular antioxidant activity experimental result indicted that cycloartenyl ferulate could alleviate H2O2-induced oxidative stress.

Novel ruthenium(ii) and (iii) carborane complexes with diphosphine ligands and their application in radical polymerization

Ruthenium(ii) and (iii) carborane complexes containing XantPhos as a ligand were synthesized for the first time. It was shown that the reaction of the known complex exo-5,6,10-[Cl(Ph3P)2Ru]-5,6,10-(µ-H)3-10-H-nido-7,8-C2B9H8 with a 10% molar excess of XantPhos in benzene at 80 °C leads to a new closo-ruthenacarborane 3-Cl-3,3-[x2-XantPhos]-closo-3,1,2-RuC2B9H11, which can be easily converted to the corresponding acetonitrile complex 3-CH3CN-3,3-[x2-XantPhos]-closo-3,1,2-RuC2B9H11 by the reaction with isopropylamine in a dichloromethane—acetonitrile solvent mixture at 40 °C. These compounds, as well as previously synthesized ruthenium(ii) carborane complexes, were used as a basis for new catalyst systems allowing to conduct controlled radical polymerization at high rates even at low catalyst loading. The specific features of methyl methacrylate polymerization under the action of the indicated catalyst systems were considered and the mechanism of the process was investigated.

Reversible Interconversion of a Static Metallosupramolecular Cage Assembly into a High-Speed Rotor: Stepless Adjustment of Rotational Exchange by Nucleophile Addition.

The self-assembled cage ROT-1 was prepared from the pyridine-terminated rotator 1, the phenanthroline-appended stator 2, DABCO, and copper(I) ions in a ratio of 1:1:1:4. This four-component assembly is held together by two pyridine→[Cu(phenAr2)]+ as well as two DABCO→zinc porphyrin interactions (phenAr2 = 2,9-diarylphenanthroline) and does not show any motion on the NMR time scale ( k < 0.1 s-1, 298 K). However, it is converted to the fast nanorotor ROT-1 xCD3CN by addition of CD3CN [ x = (v/v)% of acetonitrile in dichloromethane] due to acceleration of both pyridine→copper(I) dissociation steps. Now the rotator is able to visit all four copper(I)-loaded phenanthroline stations of the stator. Depending on the amount of CD3CN, the exchange frequency of the nanorotor varies from 0.7 s-1 (CD3CN:CD2Cl2 = 1:29) to 8000 s-1 (CD3CN:CD2Cl2 = 1:5) at 25 °C. When iodide (I-) is added to the static assembly ROT-1, the rotational speed increases even more drastically ( k = 20 000 s-1), again due to accelerating the rate-determining pyridine→copper(I) dissociation step. In both cases, a sigmoidal relationship is established between exchange frequency and the concentration of added nucleophile (CD3CN or iodide) that suggests the presence of a cooperative effect. Reversible switching between the static assembly and fast rotor was performed several times without any decomposition of the system. In contrast, addition of the common nucleophile PPh3 to ROT-1 does not increase the rotational speed, a finding that is explained on thermodynamic grounds.

Electrochemical deposition of poly[ethylene-dioxythiophene] (PEDOT) films on ITO electrodes for organic photovoltaic cells: control of morphology, thickness, and electronic properties

In this article, controlled changes on morphology, thickness, and band gap of poly[ethylenedioxythiophene] (PEDOT) polymer films fabricated by electrochemical polymerization (potentiostatically) are analyzed. Electropolymerization of the monomer ethylenedioxythiophene (EDOT) was carried out on indium tin oxide (ITO) electrodes, in different dry organic electrolytic media, such as acetonitrile, acetonitrile–dichloromethane, and toluene–acetonitrile mixtures. It was found that electropolymerization kinetics can be controlled by changing the polarity of the electrolytic media, and kinetics is slower for those with low polarity. This fact combined with an accurate control of EDOT monomer concentration and electropolymerization at Epeak/2 potential, allows to control the morphology and thickness of the electropolymerized PEDOT films (E-PEDOT:ClO4); toluene/ACN (4:1, v/v) and [EDOT] = 0.3 mM gave the best films for application in organic photovoltaic (OPV) cells. The performance of the E-PEDOT:ClO4 films was tested on ITO electrodes as anode buffer layer in OPV cells with the configuration ITO/E-PEDOT:ClO4/P3HT:PC61BM/Field’s metal, where Field’s metal (cathode) is a eutectic alloy that lets to fabricate OPV devices easily and in a fast and economical way at free vacuum conditions. The performance of these devices was compared with an OPV device constructed with a buffer layer anode, prepared using the classical spin coating of PEDOT:PSS on ITO. Results showed that OPV cells fabricated with E-PEDOT:ClO4 have a slightly increased PV performance.

Pesticides in fine airborne particles: from a green analysis method to atmospheric characterization and risk assessment

The intensive use of pesticides such as herbicides, insecticides, fungicides and acaricides has been lead to ubiquitous contamination, being present not only in soils, water bodies and/or crops, but also in the atmosphere. Considering the massive amount of pesticides employed globally, together to their persistence, this may be an important concern regarding air quality and human health worldwide. In the present study we developed a green sensitive sample preparation method for determination of nine organophosphates, two pyrethroids, one carbamate, and one strobirulin in PM2.5 collected in a tropical coastal area in the Southern Hemisphere for the first time. Extraction of PM2.5 sample masses, as low as 206 µg, were performed in a miniaturized device using 500 μL of a mixture containing 18% acetonitrile in dichloromethane followed by sonication for 23 minutes and injection into GC-MS. A total of 12 pesticides were identified and quantified successfully, among them, eight banned pesticides. A risk assessment exposure and cancer risk for possible carcinogenic pesticides (bifenthrin, malathion, parathion and permethrin) were performed for exposure of adults, children and infants. Hazard Quotient and cumulative exposure for organophosphate and pyrethroid pesticides were less than 1, showing that cumulative risk is within acceptable range.

Multiresidue Analysis of Five Neonicotinoid Insecticides and Their Primary Metabolite in Cucumbers and Soil Using High-Performance Liquid Chromatography with Diode-Array Detection.

A sensitive, selective, and validated HPLC-diode-array detection method was developed for the simultaneous determination of five neonicotinoid insecticides-acetamiprid, imidacloprid, nitenpyram, flonicamid, and thiacloprid-and their primary metabolite, 6-chloronicotinic acid, in cucumbers and soil based on the quick, easy, cheap, effective, rugged, and safe (QuEChERS) technique as a pretreatment procedure. In the QuEChERS procedure, cucumber samples were extracted with acetonitrile and cleaned using C18, whereas soil samples were extracted with an acetonitrile-dichloromethane mixture (1 + 2). The HPLC conditions were optimized by separating neonicotinoids using an acetonitrile-water mixture (25 + 75) and a Synergi Hydro RP C18 column. Matrix-matched calibration standards were prepared in cucumber and soil to eliminate any matrix interference. RSDs were ≤9% in all recovery tests. LODs and LOQs for the five neonicotinoids were in the ranges of 0.006-0.122 and 0.018-0.366 μg/g, respectively. This method was successfully applied to determine residues, the rate of disappearance of the five neonicotinoids from cucumber and soil, and the half-lives of the neonicotinoids.

The use of chemometrics to study multifunctional indole alkaloids from Psychotria nemorosa (Palicourea comb. nov.). Part I: Extraction and fractionation optimization based on metabolic profiling

Extraction methods evaluation to access plants metabolome is usually performed visually, lacking a truthful method of data handling. In the present study the major aim was developing reliable time- and solvent-saving extraction and fractionation methods to access alkaloid profiling of Psychotria nemorosa leaves. Ultrasound assisted extraction was selected as extraction method. Determined from a Fractional Factorial Design (FFD) approach, yield, sum of peak areas, and peak numbers were rather meaningless responses. However, Euclidean distance calculations between the UPLC-DAD metabolic profiles and the blank injection evidenced the extracts are highly diverse. Coupled with the calculation and plotting of effects per time point, it was possible to indicate thermolabile peaks. After screening, time and temperature were selected for optimization, while plant:solvent ratio was set at 1:50 (m/v), number of extractions at one and particle size at ≤180μm. From Central Composite Design (CCD) results modeling heights of important peaks, previously indicated by the FFD metabolic profile analysis, time was set at 65min and temperature at 45°C, thus avoiding degradation. For the fractionation step, a solid phase extraction method was optimized by a Box-Behnken Design (BBD) approach using the sum of peak areas as response. Sample concentration was consequently set at 150mg/mL, % acetonitrile in dichloromethane at 40% as eluting solvent, and eluting volume at 30mL. Summarized, the Euclidean distance and the metabolite profiles provided significant responses for accessing P. nemorosa alkaloids, allowing developing reliable extraction and fractionation methods, avoiding degradation and decreasing the required time and solvent volume.

Determination of vitamin K1 in blood serum by high-performance liquid chromatography

A method is proposed for the determination of vitamin K1 concentration in blood serum using HPLC on a Gemini C18 (250 mm × 4.6 mm ID, 5 μm) column and elution with a methanol–acetonitrile–dichloromethane (45 : 50 : 5, v/v) mobile phase at a flow rate of 1 mL/min and detection at 248 nm. The limit of detection for the vitamin is 0.5 ng/mL and relative error is 19%. The volume of sample in the column is 100 μL. The method has been applied to the evaluation of vitamin K1 deficit in a human body in medical practice.

A simple, comprehensive, and miniaturized solvent extraction method for determination of particulate-phase polycyclic aromatic compounds in air

The method allowed simultaneous characterization of PAHs, nitro-PAHs and quinones in atmospheric particulate matter. This method employs a miniaturized micro-extraction step that uses 500μL of an acetonitrile–dichloromethane mix and instrumental analysis by means of a high-resolution GC–MS. The method was validated using the SRM1649b NIST standard reference material as well as deuterated internal standards. The results are in good agreement with the certified values and show recoveries between 75% and 145%. Limit of detection (LOD) values for PAHs were found to be between 0.5pg (benzo[a]pyrene) to 2.1pg (dibenzo[a,h]anthracene), for nitro-PAHs ranged between 3.2pg (1-nitrobenzo[e]pyrene) and 22.2pg (3-nitrophenanthrene), and for quinones ranged between 11.5pg (1,4-naphthoquinone) and 458pg (9,10-phenanthraquinone). The validated method was applied to real PM10 samples collected on quartz fiber filters. Concentrations in the PM10 samples ranged from 0.06 to 15ngm−3 for PAHs, from<LOD to 69.4ngm−3 for nitro-PAHs, and from 0.27 to 115ngm−3 for quinones. This procedure was found to be precise, accurate and suitable to be employed for determination of polycyclic aromatic compounds in airborne particles from both polluted and non-polluted atmospheres.

Low band gap polymers synthesized by electrochemical polymerization for electrochromic devices

Two novel monomers, 10,13-bis(4-octyl-2-thienyl)-dibenzo[a,c]phenazine (OTBP) and 10,13-bis(4-octyl-2-thienyl)-dipyrido[3,2-a:2′,3′-c]phenazine (OTPP), were synthesized. Electrochemical polymerization of OTPP and OTBP were carried out in acetonitrile/dichloromethane (ACN/DCM) solvent mixture containing tetra-n-butylammoniumhexafluorophosphate (TBAPF6), and electrochromic properties of POTPP and POTBP are described. Furthermore, the effects of structural difference on electronic properties of electron acceptor units, electrochemical and electrochromic properties of two resulting polymers were examined. The results showed that the OTBP was polymerized at lower potential than OTPP, and this is because OTPP contains two more N atoms in electron acceptor unit and its electron withdrawing ability is stronger than that of OTBP. The UV-Vis-NIR spectra analysis revealed that the POTPP film has three absorption bands centered at 570 nm, 925 nm and 1581 nm, and the POTBP film has two absorptions at 566 nm and 1009 nm, respectively. The electrochromic properties indicated that the two polymer films exhibits reasonable optical contrasts and response time with low band gaps, defined as the onset of the absorption band at 570 nm for POTPP and the absorption band at 566 nm for POTBP, as 1.74 eV for POTPP and as 1.69 eV for POTBP.

A Three-Phase Solvent System in High-Speed Counter-Current Chromatographic for the Separation and Purification of Bioactive Constituents from Acanthus ilicifolius

A new three-phase solvent system, n-hexane–acetonitrile–dichloromethane–water–ethyl acetate (5:5:1:5:1.5, v/v/v/v/v) was developed for the high-speed counter-current chromatographic (HSCCC) separation and purification of five bioactive constituents, syringic acid (1), vomifoliol (2), vanillic acid (3), 6-hydroxy-2-benzoxazolinone (4), and 2-benzoxazolinone (5), from Acanthus ilicifolius.

Estimation of Lipophilicity of Some Polyoxygenated Steroids by the Means of Normal-Phase Thin-Layer Chromatography

Unmodified silica gel in combination with two mobile phases, acetone–n-hexane and acetonitrile–dichloromethane, was used in order to evaluate the capability of normal-phase (NP) chromatography on bare silica gel in estimation of lipophilicity of some polyoxygenated steroids. Soczewinski equation coefficients were employed as a measure of lipophilicity. The RM0 values obtained in NP-systems were correlated with those derived by extrapolation from reversed-phase (RP) systems. In addition, retention data, i.e., lipophilicity parameters determined in NP systems were compared with logP values calculated by use of several commercial computer programs. The results showed that chromatographic parameters RM0, and m obtained in NP system consisted of silica gel as stationary phase and acetone–n-hexane as mobile phase, are acceptable as the measures of lipophilicity of polyoxygenated steroids. The mechanism of retention was discussed.

Conductometric Studies of the Thermodynamics for Complexation of 1,1-Diethyl-3-(4-methoxybenzoyl)thiourea and Cobalt(II) Cation in Aqueous Binary Mixtures of Polar Organic Solvents

The complexation reaction of Co2+ (M) and the 1,1-diethyl-3-(4-methoxybenzoyl)thiourea (L) ligand in the binary mixtures: acetonitrile–dimethylsulfoxide (MeCN–DMSO), acetonitrile–dichloromethane (MeCN–DCM), water–dimethylsulfoxide (H2O–DMSO) and acetonitrile–water (MeCN–H2O), was investigated using a conductometric method at temperatures of (288.15, 298.15, 308.15 and 318.15) K. In all cases, the conductance data showed that the stoichiometry of the neutral complex formed between M and L is 1:3 [M:3L], which indicates that the Co2+ ion was oxidized to Co3+ during the reaction. The best estimate of the stability constant, log10 K f, = 3.31 for the [CoL3] complex, was observed with the MeCN–DCM (20:80 %) binary mixture at 298.15 K. The values of the thermodynamic parameters (i.e., $$ \Delta G_{\text{c}}^{^\circ } $$ and $$ \Delta S_{\text{c}}^{^\circ } $$ ) for the formation of the [CoL3] complex were obtained from the temperature dependence of the stability constant via van’t Hoff plots. The results show that, in most cases, the [CoL3] complexes are enthalpy destabilized but entropy stabilized. The values and signs of the thermodynamic parameters are affected by the nature and composition of the mixed solvents but are independent of the temperature.

Thermodynamic study of complex formation between Kryptofix-5 and Sn2+ in several individual and binary non-aqueous solvents using a conductometric method

The complex formation between 1,13-bis(8-quinolyl)-1,4,7,10,13-pentaoxatridecane (Kryptofix-5) and Sn2+ ions was studied in pure acetonitrile (AN), dimethylformamide (DMF), 1,4-dioxane (DOX), and methanol (MeOH) and in acetonitrile-1,4-dioxane (AN-DOX), acetonitrile-dichloromethane (AN-DCM), acetonitrile-methanol (AN-MeOH), and acetonitrile-dimethylformamide (AN-DMF) binary mixed solvent solutions at different temperatures using conductometric method. 1: 1 [ML] complex is formed between the metal cation and ligand in most solvent systems but in the cases of AN-MeOH (MeOH = 90 mol %) binary mixture and in pure MeOH a 2: 1 [M2L] complex was observed, that is the stoichiometry of complexes may be changed by the nature of the medium. The stability order of the (Kryptofix-5·Sn)2+ complex in the studied binary mixed solvent solutions at 25°C was found to be AN-DOX > AN-DCM > AN-MeOH > AN-DMF and in the case of pure solvents at 25°C the sequence was the following: AN > DMF > DOX. A non-linear behavior was observed for changes of logK f of (Kryptofix-5·Sn)2+ complex versus the composition of the binary mixed solvents, which was explained in terms of solvent-solvent intractions and also by the preferential solvation of the f species involved in the complexation reaction. The values of standard enthalpy changes (ΔH c ○ ) for complexation reactions were obtained from the slope of the Van’t Hoff plots and the changes in standard entropy (ΔS c ○ ) were calculated from the relationship ΔG c, 298.15 ○ = ΔH c ○ − 298.15ΔS c ○ . The results show that in most cases, the (Kryptofix-5·Sn)2+ complex is both enthalpy and entropy stabilized.

A complexation study of 15-crown-5 with Co2+ cation in some pure and mixed organic solvents

The stability constant (log Kf) and the thermodynamic parameters (free energies, enthalpies, and entropies) of the complexation of Co2+ cation with 15-crown-5 (15C5) in acetonitrile-methanol (AN/MeOH), acetonitrile-nitrobenzene (AN/NB), acetonitrile-dichloromethane (AN/DCM) and acetonitrile-1,2-dichloroethane (AN/DCE) binary solvent solutions were calculated from the experimental conductance data at different temperatures. The complexation behavior of the crown ether used in these media was discussed in view of the estimated parameters. In all solvent systems, 15-crown-5 formed a 1: 1 complex with Co2+ cation. The stability order of (Co-15C5)2+ complex in the binary mixed solvents at 25°C was found to be: AN/NB > AN/DCM ≈ AN/DCE > AN/MeOH. In most cases, a non-linear relationship was observed for changes of log Kf of (Co-15C5)2+ complex versus the composition of the binary mixed solvent systems. The experimental results show that the standard thermodynamic parameters of the complexation process change with the nature and composition of the binary solvent solutions.

Conformational Polymorphism and Isomorphism of Molecular Rotors with Fluoroaromatic Rotators and Mestranol Stators

We report the synthesis and characterization of four molecular rotors 2, 3, 4, and 5 containing 2-fluoro-, 2,3-difluoro-, 2,5-difluoro-, and 2,3,5,6-tetrafluoro-substituted 1,4-phenylene rotators, respectively, that are axially linked through the triple bonds of rigid mestranol (3-methoxy-17α-ethynylestradiol) stators. Crystallization experiments using solvent mixtures of hexanes–ethyl acetate and acetonitrile–dichloromethane gave rise to polymorphic, pseudopolymorphic, and isomorphic crystals. Whereas two solids were obtained for compound 2, four were indentified in the cases of compounds 3 and 4, and three for compound 5. The 13 solid forms were characterized by single-crystal and powder X-ray diffraction, infrared spectroscopy, differential scanning calorimetry, and thermogravimetric analysis. Two sets of isomorphous structures were obtained for solvated and solvent-free structures of compounds 3–5. While polymorphic behavior of these compounds arises from their conformational freedom in solution, their isomorphism arises from the closely isosteric relation that exists between hydrogen and fluorine atoms.

The separation of flavonoids from Pongamia pinnata using combination columns in high-speed counter-current chromatography with a three-phase solvent system

The mangrove plant Pongamia pinnata (Leguminosae) is well known as a plant pesticide. Previous studies have indicated that the flavonoids are responsible of the biological activities of the plant. A new high-speed counter-current chromatography (HSCCC) method for the separation of three flavonoids, karanjin (1), pinnatin (2), and pongaflavone (3), from P. pinnata was developed in the present study. The lower and intermediate phase (LP and IP) of a new three-phase solvent system, n-hexane–acetonitrile–dichloromethane–water, at a volume ratio of 5:5:1:5, were used as the stationary phases, while the upper phase (UP) was used as the mobile phase, and the volume ratio between the stationary phases in the CCC column could be tuned by varying the initial pumped volume ratio of the stationary phases. The CCC columns containing all three phases of the solvent system were considered combination columns. According to the theories of combination column, it is possible to optimize the retention time of the target compounds by varying the volume ratio of the stationary phases in the HSCCC combination columns, as well as the suitable volume ratios of the stationary phases for the separation of the target compounds were predicted from the partition coefficients of the compounds in the three-phase solvent system. Then, three HSCCC separations using the combination columns with initial pumped LP:IP volume ratios of 1:0, 0.9:0.1, and 0.7:0.3 were performed separately based on the prediction. Three target compounds were prepared with high purity when the initial pumped volume ratio of the stationary phases was 0.9:0.1. The baseline separation of compounds 2 and 3 was achieved on the combination column with an initial pumped volume ratio of 0.7:0.3. Furthermore, the three experiments clearly demonstrated that the retentions and resolutions of the target compounds increased with an increasing volume ratio of IP, which is consistent with the prediction for the retention times for the solutes on combination columns. The method proposed here reduces the need for solvent selection compared with the conventional method and may have broad potential applicability in the preparation of natural products.

Chemical Constituents of Cinnamomum rupestre and Cinnamomum nanophyllum

Cinnamomum rupestre Kosterm. (Lauraceae) is an endemic species in the Philippines originally discovered in Palawan [1], while Cinnamomum nanophyllum Kosterm. is a widespread endemic species in the country. In Cebu, the local people collect and prepare the leaves and the bark as a decoction for relief of stomach and abdominal ailments. There are no reported chemical and biological studies on C. rupestre and C. nanophyllum. These are two of the 21 species of Cinnamomum found in the Philippines, of which 16 are known to be endemic [1]. This study was conducted as part of our research on the chemical constituents of Cinnamomum species found in the Philippines. We earlier reported the isolation of eugenol and safrole from the bark, polyprenol from the leaves, and trilinolein from the roots of Cinnamomum cebuense [2]. Silica gel chromatography of the dichloromethane (DCM) extract of the bark of C. rupestre afforded 4 -hydroxy5,7,3 -trimethoxyflavan-3-ol (1), 4-hydroxy-3-methoxycinnamaldehyde (2), and 4-allyl-2-methoxyphenol or eugenol (3), while the leaves yielded -sitosterol (4). The structure of 1 was elucidated by extensive 1D and 2D NMR spectroscopy and confirmed by comparison of its 13C NMR data with those reported in the literature for 4 -hydroxy-5,7,3 -trimethoxyflavan-3-ol (1) [3]. The structures of 4-hydroxy-3-methoxycinnamaldehyde (2) [4], 4-allyl-2-methoxyphenol or eugenol (3) [5], and -sitosterol (4) [6] were confirmed by comparison of their 13C NMR data with those reported in the literature. The DCM extract of the bark of C. nanophyllum afforded 1, 2, 4, and trilinolein (5), while the leaves yielded 4. The structure of trilinolein (5) [7] was confirmed by comparison of its 13C NMR data with those reported in the literature. To the best of our knowledge, this is the first report on the chemical constituents of C. rupestre and C. nanophyllum. The bark and leaves of Cinnamomum rupestre and Cinnamomum nanophyllum were collected from Hagnaya, Carmen, Cebu, Philippines in June 2010. Voucher specimens of Cinnamomum rupestre and Cinnamomum nanophyllum were authenticated by one of the authors (EMGA) and deposited in the De La Salle University-Manila Herbarium (DLSU-3101 and DLSU-3102, respectively). The air-dried bark and leaves of C. rupestre were chopped into small pieces and then air dried. The air-dried bark (371.8 g) and leaves (89.1 g) were soaked in DCM for 3 days and then filtered. The filtrates were concentrated under vacuum to afford the crude extracts of bark (16.6 g) and leaves (7.0 g), which were fractionated by silica gel chromatography using increasing proportions of acetone in DCM (10% increment by volume) as eluents. The DCM fraction from the bark was rechromatographed in petroleum ether (7 ) to afford 3 (28 mg). The 30% acetone in DCM fraction was rechromatographed with 12.5% ethyl acetate in petroleum ether as eluent to afford 2 (18 mg). The 40% acetone in DCM fraction was rechromatographed (5 ) in DCM, then (8 ) in diethyl ether–acetonitrile–DCM (0.5:0.5:9) to afford 1 (12 mg). The DCM fraction from the leaves was rechromatographed (3 ) in 5% ethyl acetate in petroleum ether to afford 4 (10 mg). The air-dried bark and leaves of C. nanophyllum were chopped into small pieces and then air dried. The air-dried bark (493.5 g) and leaves (49.5 g) were soaked in DCM for three days and then filtered. The filtrates were concentrated under vacuum to afford the crude extracts of bark (8.0 g) and leaves (9.5 g), which were fractionated by silica gel chromatography using increasing proportions of acetone in DCM (10% increment by volume) as eluents. The crude DCM extract of the bark of C. nanophyllum was chromatographed in increasing proportions of acetone in DCM at 10% increment as eluents. The 10%