Preparative isolation and purification of sinomenine from Sinomenium acutum by centrifugal partition chromatography

Preparative centrifugal partition chromatography (CPC) was successfully carried out for the separation of sinomenine from the methanolic extract of Sinomenium acutum stems and rhizomes. The optimum two-phase solvent system of CPC was composed of n-hexane/ethyl acetate/methanol/water at a volume ratio of 1:6:2:8 (v/v/v/v) with 0.1% triethylamine (TEA). Preparative CPC yielded 44.3 mg of sinomenine from 400 mg of MeOH extract with a purity of 96.9%.     

Supercritical fluid extraction of sinomenine from Sinomenium acutum (Thumb) Rehd et Wils

Supercritical carbon dioxide, with and without a methanol modifier, was used to extract sinomenine from Sinomenium acutum (Thumb) Rehd et Wils. Sinomenine determinations were carried out using high-performance liquid chromatography (HPLC). The results show that the yield obtained after 2.5 h extraction with methanol-modified supercritical carbon dioxide was the highest (7.47 mg/g), while that obtained with only supercritical carbon dioxide was the lowest (0.17 mg/g). The recovery obtained with supercritical carbon dioxide, with and without a methanol modifier, could not be increased greatly by the method of the alkalinization of sample. Higher recoveries were obtained than extraction using methanol in Soxhlet extractor.     

毛细管电泳高频电导法测定青风藤药材及制剂中青藤碱的含量

青藤碱（Sinomenine)是具有镇痛、消炎、怯风除湿、活血化瘀、增强免疫和抗癌等作用的生物碱单体(结构式见Scheme 1),它存在于防己科植物青风藤和毛青藤的干燥藤茎中,在临床上,已有正青风痛宁片、盐酸青藤碱注射液和毛青藤总片等制剂,用于治疗各种急慢性风湿痛、肿胀以及心率失常等病症．青藤碱含量测定常用非水滴定法,但该法专属性差,实验结果易偏高．此外,     

Identification of Sinomenine from Sinomenium actum and Its Simultaneous Quantitation by GC–MS and Non-Aqueous CE

In the present study, the qualitative and quantitative analysis of alkaloids in the stem and root of Sinomenium acutum (S. acutum) is presented by gas chromatography-mass spectrometry (GC–MS) and non-aqueous capillary electrophoresis (NACE). The extract of alkaloids in S. acutum was examined by GC–MS and the major alkaloids were identified. Sinomenine (SIN) was found as the principal alkaloid in the extracts (about 84.38%). The quantitation of SIN was then accomplished by GC–MS and NACE with diode array detection. NACE was selected in order to use a running buffer fully compatible with samples in organic solvent. Optimum separation was achieved with a fused-silica capillary column and a running buffer containing 80 mM ammonium acetate, 2.0% acetic acid and 20% (v/v) acetonitrile in methanol medium. The applied voltage was 22 kV. The different selectivity displayed by these techniques allowed different separation profiles that could be useful in phytochemical characterization of the sample. The GC–MS and NACE methods were successfully validated and applied for the quantitation of SIN in S. acutum.     

Pharmacokinetics and penetration into synovial fluid of systemical and electroporation administered sinomenine to rabbits

Sinomenine is an anti‐rheumatoid arthritis (RA) drug derived from the Sinomenium acutum. The major site of RA treatment is within the synovial compartment. However, the pharmacokinetic and penetration into synovial fluid (SF) of sinomenine have not been reported. In our study, the pharmacokinetics and penetration into SF of systemic and electroporation administered sinomenine were investigated by microdialysis incorporated with HPLC‐MS/MS. Sinomenine went into plasma and SF more rapidly with higher peak concentration (C_max) by intramuscular injection compared with oral administration. The area under the concentration–time graph (AUC_0–∞) of intramuscularly injected sinomenine was 1,403,294.75 ± 125,534.567 ng min/mL in plasma and 456,116.37 ± 62,648.36 ng min/mL in SF, which were equivalent with those for an oral dose. These results indicated that equal amounts of sinomenine could penetrate into SF by the two administration routes, and the permeation ratios were approximately 1:3. The AUC_0–∞ and C_max were lower with electroporation compared with systemic administration, but the C_SF/C_Plasma (concentration of sinomenine in SF vs that of plasma) at 90, 120, 150, 180, 240 and 480 min by electroporation was 3‐ to 10‐fold higher relative to systemic administration. This illustrated that sinomenine can be targeted into joints by electroporation, and electroporation is a potential technique for sinomenine's transdermal delivery. Copyright © 2014 John Wiley & Sons, Ltd.     

Sinoracutine, a novel skeletal alkaloid with cell-protective effects from Sinomenium acutum

From the stems of Sinomenium acutum, sinoracutine (1) has been isolated, an alkaloid with a structurally novel skeletal framework, whose structure has been established by spectral and single crystal X-ray diffraction analysis. In vitro experiments show that sinoracutine increases cell viability against Hydrogen peroxide-induced oxidative injury.     

青风藤有效化学成分与环氧合酶的对接研究

将青风藤中23个有效化学成分与COX-2酶对接,有四个化合物具有较低的结合自由能,其中以青藤碱为最低.进一步将这四个化合物与COX-1酶对接,发现这四个化合物与COX-1酶结合能力较弱,预示这四个化合物具有选择性抑制COX-2酶的能力.对青藤碱和COX-2以及COX-1酶的结合模式进行分析,发现青藤碱主要结合于COX-2酶的S′口袋,而COX-1酶的S′结合口袋中第523号残基由COX-2酶中的Val523变成了体积较大的Ile523,使得COX-1酶的S′结合口袋相对COX-2酶的结合口袋要小,从而导致青藤碱分子不能进入COX-1酶S′结合口袋.这成功解释了青藤碱选择性抑制COX-2酶的原因,与早期有关文献报道的实验结果相吻合,充分表明了对接模型的合理性,青藤碱等化合物可作为设计COX-2酶选择性抑制剂的先导化合物.     

Crystal Structure of 8-Demethoxyrunanine

A new hasubanane-type alkaloid, 8-demethoxyrunanine, was isolated from Sino- menium acutum and characterized by melting point, HREIMS, 1H NMR, and X-ray diffraction analysis. X-ray diffraction reveals that the title compound crystallizes in the orthorhombic system, space group P212121 with a = 7.308（1）, b = 21.742（5）, c = 22.893（4）A, V = 3637.5（11） A^3, Z = 8, Dx = 1.254 g/cm^3, F（000） = 1472, μ（MoKα） = 0.087 mm^-1, the final R = 0.0438 and wR = 0.0575 for 4497 independent reflections with Rint = 0.0192 and 2091 observed reflections with I 〉 2σ（I）. Four rings （ring A： one benzene ring, ring B： one hexagon carbon ring in a half-chair conformation, ring C： one hexagon carbon ring with α,β-unsaturated ketone segment （-CR2=CR1-C=O） in a screw-boat conformation, and ring D： one nonplanar tetrahydropyrrole） form a hasubanane-type alkaloid.     

New alkaloids sinomacutines A–E,and cephalonine-2-O-β-d-glucopyranoside from rhizomes of Sinomenium acutum

Phytochemical investigation on the rhizomes of Sinomenium acutum led to the isolation of new alkaloids sinomacutines A–E (1–5), and cephalonine-2-O-β-d-glucopyranoside (6), along with known tetrahydroisoquinoline alkaloids (7–9). Among them, sinomacutines A–C were new bistetrahydroisoquinoline alkaloids with morphinane–proaporphine and morphinane–benzyltetrahydroisoquinoline types, and coupled with unique C–CH₂–N unit. Their structures with absolute configuration were elucidated on the basis of 1D and 2D NMR, and ECD spectra analysis. The new compounds were evaluated for their anti-inflammatory and cytotoxic activities, and sallisonine B (2) showed moderate ability to inhibit NO production of LPS-stimulated RAW264.7 macrophages, as well as sallisonine A (1) showed weak inhibitory effects against five cancer cell lines.     

青藤碱和黄芩甙的极谱特性

用单扫示小极谱法研究了青藤碱和黄芩甙的电化学行为，青藤碱在０．１ｍｏｌ／Ｌ Ｎａ２Ｂ４Ｏ７中有两个还原峰，峰Ｐ１和Ｐ２的电位分别为－１．４５Ｖ和－１．６７Ｖ，峰Ｐ１的峰电流与浓度在０．０６－１．８ｍｇ／Ｌ和２．０－３４．６ｍｇ／Ｌ范围内呈线性关系，检测限为０．０２ｍｇ／Ｌ，在０．０５ｍｏｌ／Ｌ棕檬酸钠＋０．２ｍｏｌ／Ｌ ＨＣｌ底液中，黄芩甙于－１．１４Ｖ（Ｐ３０和－１．３２Ｖ（Ｐ４）处产生两还原峰     

Application of nonaqueous capillary electrophoresis for quantitative analysis of quinolizidine alkaloids in Chinese herbs

An easy, rapid method for simultaneous determination of tetrandrine (TET), fangchinoline (FAN), sinomenine (SIN) and tetrahydropalmatine (TEP) in Chinese herbs was developed by nonaqueous capillary electrophoresis without pretreatment for the first time. Optimum separation was achieved with a fused-silica capillary column ( i.d.) and a running buffer containing 50 mM ammonium acetate, 1.0% acetic acid and 20% acetonitrile in methanol medium. The applied voltage was 20.0 kV. The analytes were detected by UV at 214 nm. The effects of concentration of ammonium acetate, acetic acid and organic modifier on electrophoretic behavior of the analytes were studied. Regression equations revealed linear relationships (correlation coefficients: 0.9991-0.9999) between the peak area of each analyte and the concentration. The levels of analytes in Stephania tetrandra S. Moore, Rhizoma corydalis and Sinomenium acutum Rehd. et. Wils were easily determined with recoveries ranging from 96 to 107%.     

Chemical enhancement by nanomaterials under X-ray irradiation

We report here a new phenomenon of dynamic enhancement of chemical reactions by nanomaterials under hard X-ray irradiation. The nanomaterials were gold and platinum nanoparticles, and the chemical reaction employed was the hydroxylation of coumarin carboxylic acid. The reaction yield was enhanced 2000 times over that predicted on the basis of the absorption of X-rays only by the nanoparticles, and the enhancement was found for the first time to depend on the X-ray dose rate. The maximum turnover frequency was measured at 1 × 10(-4) s(-1) Gy(-1). We call this process chemical enhancement, which is defined as the increased yield of a chemical reaction due to the chemical properties of the added materials. The chemical enhancement described here is believed to be ubiquitous and may significantly alter the outcome of chemical reactions under X-ray irradiation with the assistance of nanomaterials.     

Determination of sinomenine in Sinomenium acutum by capillary electrophoresis with electrochemiluminescence detection

A Ru(bpy)₃²⁺-based electrochemiluminescence (ECL) detection coupled with capillary electrophoresis (CE) has been established for the determination of sinomenine for the first time. Optimum separation was achieved with a fused-silica capillary column (50 cm × 25 μm i.d.) and a background electrolyte of 50 mM sodium phosphate (pH 5.0) at a separation voltage of 15 kV. The content of sinomenine was detected by ECL at the detection voltage of 1.15 V (versus Ag/AgCl) with 5 mM Ru(bpy)₃²⁺ in 75 mM phosphate solution (pH 8.0) when a chemically modified platinum electrode by europium(III)-doped prussian blue analogue (Eu-PB) was used as a working electrode. Under the optimized conditions, the ECL intensity was in proportion to sinomenine concentration in the range from 0.01 to 1.0 μg mL⁻¹ with a detection limit of 2.0 ng mL⁻¹ (3σ). The relative standard derivations of migration time and ECL intensity were 0.93 and 1.11%, respectively. The level of sinomenine in Sinomenium acutum Rehd. et Wils was easily determined with recoveries between 98.6 and 102.7%.     

Structure of a novel epidithiodioxopiperazine, emethallicin A, a potent inhibitor of histamine release from Emericella heterothallica

A new epidithiodioxopiperazine derivative, emethallicin A (1), was isolated along with ergosterol from the mycelial extract of the heterothallic fungus, Emericella heterothallica (mating type A). The structure of emethallicin A (1) was established on the basis of chemical and spectroscopic investigations and finally by chemical transformation to apoaranotin (11). Emethallicin A (1) has the same basic skeleton as apoaranotin (11), with C6-C2 carboxylic acid (mandelic and phenylacetic acids) diester moieties, and showed a potent inhibitory activity on histamine release from mast cells.     

Complete assignments of (1)H and (13)C NMR spectroscopic data for two new triterpenoid saponins from Ilex pernyi

Ilexpernoside A and ilexpernoside B, two new pentacyclic C(4)-nortriterpenoid saponins, were isolated from the leaves of Ilex pernyi Franch. Their chemical structures were determined by MS, NMR spectroscopy and chemical analysis. Complete assignments of the (1)H and (13)C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, (1)H-(1)H COSY and NOESY).     

Complete assignments of 1H and 13C NMR spectroscopic data for two new triterpenoid saponins from Ilex hainanensis

Two novel unsaturated E-ring pentacyclic triterpenoid saponins, ilexhainanoside A and ilexhainanoside B, were isolated from the leaves of Ilex hainanensis. Their chemical structures were determined by MS, NMR spectroscopy and chemical analysis. Complete assignments of the 1H and 13C NMR spectroscopic data were achieved by 1D and 2D NMR experiments (HSQC, HMBC, ROESY and 1H-1H COSY).     

Rehybridization and the Chemical Reaction: POAV and 3D-HMO Analysis

In this study chemical reactions are examined from a new standpoint — rehybridization. With the possible exception of oxidation-reduction processes, all chemical reactions involve rehybridization. Bondmaking and bond-breaking processes are nothing more than changes in hybridization. ln the present study two categories of pericyclic reactions are examined with the π-orbital axis vector (POAV) analysis and-the three-dimensional (3D) HMO theory. The first makes use of a series of related molecular structures derived from X-ray crystallography which provide information on the reaction pathway between the valence tautomers 1,6-methano[10]annulene (1a) and the bisnorcaradiene (1b). The second category involves structures calculated by Dewar and coworkers along the minimum energy reaction pathway of the Cope rearrangement of 1,5-hexadiene (2). The analysis provides a clear delineation of the structural and electronic changes which accompany chemical reaction. The sense of the π-orbital axis vectors at the bridgehead atoms is shown to be the determining feature in dividing the potential surface of 1 between bridged-[10]annulene (1a) and bisnorcaradiene (1b).     

Two new steroidal glycosides from Ophiopogon japonicus

Two new C27-steroidal glycosides were isolated from the fibrous roots of Ophiopogon japonicus. The spectral analysis and chemical evidence revealed their chemical structures to be （25R）-spirost-5,14-dien-3β-yl-O-α-L-rhamnopyranosyl-（1 → 2）-β-D- xylopyranosyl-（1 → 4）-β-D-glucopyranoside （1） and ophiogenin 3-O-β-D-glucopyranoside （2）.     

Toward direct determination of conformations of protein building units from multidimensional NMR experiments. V. NMR chemical shielding analysis of N-formyl-serinamide,a model for polar side-chain containing peptides

Knowledge of chemical shift-structure relationships could greatly facilitate the NMR chemical shift assignment and structure refinement processes that occur during peptide/protein structure determination via NMR spectroscopy. To determine whether such correlations exist for polar side chain containing amino acid residues the serine dipeptide model, For-L-Ser-NH(2), was studied. Using the GIAO-RHF/6-31+G(d) and GIAO-RHF/TZ2P levels of theory the NMR chemical shifts of all hydrogen ((1)H(N), (1)H(alpha), (1)H(beta1), (1)H(beta2)), carbon ((13)C(alpha), (13)C(beta), (13)C') and nitrogen ((15)N) atoms have been computed for all 44 stable conformers of For-L-Ser-NH(2). An attempt was made to establish correlation between chemical shift of each nucleus and the major conformational variables (omega(0), phi, psi, omega(1), chi,(1) and chi(2)). At both levels of theory a linear correlation can be observed between (1)H(alpha)/phi, (13)C(alpha)/phi, and (13)C(alpha)/psi. These results indicate that the backbone and side-chain structures of For-L-Ser-NH(2) have a strong influence on its chemical shifts.     

Synthesis and application in polypropylene of a novel of phosphorus-containing intumescent flame retardant

A novel phosphorus-containing triazine oligomer poly(2-morpholinyl-4-penta-erythritol phosphate-1,3,5-triazine) (PMPT) was synthesized as a kind of tri-component intumescent flame retardant (IFR). The chemical structure of PMPT was characterized by FTIR, 1H-NMR and 31P-NMR, and the mechanical and flammability properties of FR-PP were measured. The FTIR results showed that the expected chemical reactions had happened at each step. The 1H-NMR and 31P-NMR spectra also agreed with the chemical structure of PMPT. The slight effect of PMPT on the mechanical properties of FR-PP suggested that PMPT and PP are compatible. The high limited oxygen index (LOI) values of FR-PP revealed that PMPT was an efficient IFR and there was the synergistic effect between PMPT and ammonium polyphosphate/ pentaerythritol (APP/PER).