Morphological and Molecular Phylogenetic Data of the Chinese Medicinal Fungus Cordyceps liangshanensis Reveal Its New Systematic Position in the Family Ophiocordycipitaceae

A cordycipitoid fungus infecting Hepialidae sp. in Nepal was supposed to be identical to Cordyceps liangshanensis, originally described from southwestern China, and thus, transferred to the genus Metacordyceps or Papiliomyces in previous studies. However, our multi-gene (nrSSU-nrLSU-tef-1α-rpb1-rpb2) phylogenetic and morphological studies based on the type specimen and additional collections of C. liangshanensis revealed that the fungus belongs to the genus Ophiocordyceps (Ophiocordycipitaceae). Therefore, a new combination O. liangshanensis was made, and a detailed description of this species was provided.

Morphological and Molecular Phylogenetic Data of the Chinese Medicinal Fungus Cordyceps liangshanensis Reveal Its New Systematic Position in the Family Ophiocordycipitaceae

A cordycipitoid fungus infecting Hepialidae sp. in Nepal was supposed to be identical to Cordyceps liangshanensis, originally described from southwestern China, and thus, transferred to the genus Metacordyceps or Papiliomyces in previous studies. However, our multi-gene (nrSSU-nrLSU-tef-1α-rpb1-rpb2) phylogenetic and morphological studies based on the type specimen and additional collections of C. liangshanensis revealed that the fungus belongs to the genus Ophiocordyceps (Ophiocordycipitaceae). Therefore, a new combination O. liangshanensis was made, and a detailed description of this species was provided.

Simultaneous determination of thirteen components in Tong'an injection by LC-MS/MS / 中国中药杂志

The aim of this study was to develop a simple, sensitive ultra performance liquid chromatography mass spectrometry (UPLC-MS/MS) method for the determination of syringaresinol, N-trans-feruloyltyramine, chelerythrine chloride, sinomenine, coptisine chloride, sanguinarine, chelidonine, magnolflorine, allocryptopine, protopine, farrerol, stylopine and dihydrosanguin-arine in Tong'an injection (TAI), which could be used for the quality control of TAI. The UPLC analysis was performed on Agilent Zorbax SB-Aq column (2.1 mm×150 mm,3.5 μm), with 0.1% formic acid solution (A) -acetonitrile (B) as the mobile phase for gradient elution (0.01-2 min, 5%B; 2-8 min, 5%-30%B; 8-11 min, 30%-95%B; 11-13 min, 95%B; 13-13.1 min, 95%-5%B; 13.1-14 min, 5%B). The flow rate was 0.5 mL•min⁻¹, and the column temperature was 25 ℃; multiple reaction monitoring (MRM) was performed in electrospray ion source positive ion mode for quantitative determination. The calibration curves for the above thirteen compounds showed good linear relationship in corresponding mass concentration range (r>0.999 0). The average recovery rate of the compounds ranged from 95.70% to 104.8%, with RSD of less than 1.9%. The contents of thirteen active components in 10 batches of TAI were 0.021 2-0.029 0, 0.001 7-0.002 3, 0.000 9-0.001 3, 5.952-6.205 2, 0.195 4-0.240 5, 0.002 0-0.002 9, 0.693-0.798 2, 0.069 3-0.078 2, 0.089 29-0.102 9, 0.386 5-0.420 1, 0.014 3-0.015 9, 0.755 3-0.842 1, and 0.008 2-0.011 2 g•L⁻¹ respectively. Methodology validation proved that this method was simple, rapid, sensitive and accurate, which can be used to provide reference for the comprehensive evaluation of TAI quality. The determination results of 10 batches of TAI showed the content of each batch had no significant difference. The results may provide a basis for the quality control of TAI.

Determination of residual organic solvents and macroporous resin residues in Akebia saponin D / 中国中药杂志

According to ICH, Chinese Pharmacopoeia and supplementary requirements on the separation and purification of herbal extract with macroporous adsorption resin by SFDA, hexane, acetidine, ethanol, benzene, methyl-benzene, o-xylene, m-xylene, p-xylene, styrene, diethyl-benzene and divinyl-benzene of residual organic solvents and macroporous resin residues in Akebia saponin D were determined by headspace capillary GC. Eleven residues in Akebia saponin D were completely separated on DB-wax column, with FID detector, high purity nitrogen as the carry gases. The calibration curves were in good linearity (0.999 2-0.999 7). The reproducibility was good (RSD < 10%). The average recoveries were 80.0% -110%. The detection limit of each component was far lower than the limit concentration. The method is simple, reproducible, and can be used to determine the residual organic solvents and macroporous resin residues in Akebia saponin D.

Absorption characteristics of the total alkaloids from Mahonia bealei in an in situ single-pass intestinal perfusion assay / 中国天然药物

AIM@#To investigate the absorption characteristics of the total alkaloids from Mahoniae Caulis (TAMC) through the administration of monterpene absorption enhancers or protein inhibitors.@*METHOD@#The absorption behavior was investigated in an in situ single-pass intestinal perfusion (SPIP) assay in rats.@*RESULTS@#The intestinal absorption of TAMC was much more than that of a single compound or a mixture of compounds (jatrorrhizine, palmatine, and berberine). Promotion of absorption by the bicyclic monoterpenoids (borneol or camphor) was higher than by the monocyclic monoterpenes (menthol or menthone), and promotion by compounds with a hydroxyl group (borneol or menthol) was higher than those with a carbonyl group (camphor or menthone). The apparent permeability coefficient (Papp) of TAMC was increased to 1.8-fold by verapamil, while it was reduced to one half by thiamine. The absorption rate constant (Ka) and Papp of TAMC were unchanged by probenecid and pantoprazole.@*CONCLUSION@#The intestinal absorption characteristics of TAMC might be passive transport, and the intestinum tenue was the best absorptive site. In addition, TAMC might be likely a substrate of P-glycoprotein (P-gp) and organic cation transporters (OCT), rather than multidrug resistance protein (MRP) and breast cancer resistance protein (BCRP). Compared with a single compound and a mixture of compounds, TAMC was able to be absorbed in the blood circulation effectively.

Isolation, structure characterization and quantification of related impurities in asperosaponin VI / 中国天然药物

AIM@#To study the related impurities in asperosaponin VI bulk drug and to develop a high performance liquid chromatography (HPLC) method for the determination of asperosaponin VI and its related impurities.@*METHODS@#The related impurities were detected in asperosaponin VI bulk drug by a newly developed HPLC method, obtained by ODS column chromatography and semi-preparative HPLC methods, and the structures were elucidated by TOF-MS, IR, and NMR techniques. The HPLC method was validated according to ICH guidelines for asperosaponin VI and its related impurities.@*RESULTS@#Seven related impurities (Imp 1-7) were isolated from asperosaponin VI bulk drug. Impurity 3 was found to be a mixture of two epimers, and was first reported in the paper. The validation results showed good sensitivity, specificity, linearity (r(2) ≥ 0.997 9), precision (RSD < 5.0%), accuracy (recoveries in the range of 94.61%-106.51%) and robustness.@*CONCLUSION@#The developed HPLC method is suitable for the quality control of asperosaponin VI bulk drug.

Study on pharmacokinetics of asperosaponin VI and its active metabolite in rats / 中国中药杂志

To study the pharmacokinetics, excretion characteristics and plasma protein binding rate of asperosaponin VI (A-VI) and its active metabolite hederagenin (M1). A-VI and M1 concentrations in plasma, bile, urine and feces were determined by established LC-MS/MS to calculate the pharmacokinetic parameters. The plasma protein binding rate of A-VI was determined by equilibrium dialysis method. the double peaks was observed in the A-VI plasma concentration-time curve, after rats were orally administered with low, medium and high doses of A-VI (0.03, 0.09, 0.27 g x kg(-1)). The Cmax1 and Cmax2 of A-VI were (28.88 +/- 49.78) and (4.480 +/- 1.872) microg x L(-1), (35.19 +/- 23.53) and (22.11 +/- 16.15) microg x L(-1), (73.37 +/- 37.28) and (132.2 +/- 160.7) microg x L(-1), respectively. The AUC0-t, of A-VI were (43.21 +/- 37.32), (133.9 +/- 102.5) and (779.6 +/- 876.9) microg x h x L(-1), respectively. The t1/2 of A-VI were (3.3 +/- 0.8), (3.2 +/- 2.3) and (4.5 +/- 1.2) h, respectively. The Cmax of M1 were (16.03 +/- 9.336), (26.41 +/- 11.95) and (28.71 +/- 5.874) microg x L(-1), respectively. The AUC0-t, of M1 were (105.6 +/- 73.60), (260.0 +/-153.9) and (323.1 +/- 107.9) microg x h x L(-1), respectively. The t1/2 of M1 were (4.1 +/- 3.4), (4.4 +/- 2.3), (3.9 +/- 0.9) h, respectively. No significant gender difference was found in the in vivo pharmacokinetics of A-VI and M1. There was no accumulation of A-VI and M1 after multiple administrations of A-VI (0.09 g x kg(-1)). After oral administration of A-VI, the double peaks were also observed in biliary and urinary excretion rate-time curves for A-VI. M1 was detected in the feces samples at 6 h after oral administration. The average plasma protein binding rate of A-VI was 92. 9% in rats.

Impacts of bicyclo-monoterpene enhancers on transdermal delivery of ligustrazine / 药学学报

The purpose of this study is to investigate the impacts of bicyclo-monoterpene promoters (i.e., borneol and camphor) on the in vitro permeation of ligustrazine (LGT) through the hairless porcine dorsal skin. Fourier transform-infrared (FT-IR), scanning electron microscope (SEM) and transdermal delivery kinetics in vitro were performed to investigate the effect of the promoters on the biophysical changes to the stratum corneum (SC), the surface changes to porcine skin and the in vitro percutaneous fluxes of ligustrazine through procine skin. FT-IR results revealed that the peak shift and the decrease in the peak area with borneol were higher than those with camphor. SEM studies demonstrated that the morphological change to SC was related to the chosen enhancer. It was observed that the SC lipid extraction with borneol and camphor led to disruption of the SC and the scutella desquamation. Apparent density (AD) was utilized to describe the desquamation extent of the scutella. Percutaneous fluxes of ligustrazine through porcine skin were evaluated in vitro by the Franz-type diffusion cells. Use of borneol led to greater penetration of ligustrazine across porcine skin. It was shown that the permeation enhancement mechanism of bicyclo-monoterpenes to ligustrazine included extracting and disordering lipids which involved the shift changes in C-H stretching and H-bonding action between enhancers and cermaide. The penetration capability of the hydroxy groups in bicyclo-monoterpenes was better than that of the ketone groups.

Effects of D-limonene and L-limonene on transdermal absorption of ligustrazine hydrochloride / 药学学报

<p><b>AIM</b>To investigate the effects and permeation mechanism of D-limonene and L-limonene on transdermal delivery of ligustrazine hydrochloride (LH).</p><p><b>METHODS</b>Transdermal flux of LH through porcine skin was determined in vitro by Franz-type diffusion cells. The peak shift and peak areas of C-H stretching vibration absorption were estimated by Fourier transform-infrared (FTIR). Morphological changes in the stratum corneum (SC) treated with enhancers were observed by a scanning electron microscope (SEM) and apparent density, a new concept, was proposed to estimate the desquamated extent of SC for the first time.</p><p><b>RESULTS</b>There were no statistic difference (P > 0.05) between the transdermal fluxs of the enantiomer enhancers which were higher than those of control and azone. But the lag time of L-limonene was 2.55 times than that of D-limonene. The FTIR results revealed that the shift and decreased peak area of C-H stretching vibrations in the SC lipids were dependent on the enhancers. The enantiomers permeation enhancers, D-limonene and L-limonene, were able to perturb and extract the SC lipids to different extent. The disordering and extracting lipids activity of L-limonene was stronger than that of D-limonene. SEM studies demonstrated that the extraction of lipids was depended on the selected penetration promoters.</p><p><b>CONCLUSION</b>D-limonene was the most effective enhancer which had the greater transdermal flux of LH and the least lag time. The results showed that the permeation enhancement mechanism of the enantiomer enhancers to LH was multiple ones including disordering and extracting the SC lipids and probably including stereoselective mechanism.</p>

Studies on the constituents in rhizome of Homalomena occuta / 中国中药杂志

<p><b>OBJECTIVE</b>Bio-active constituents were expected to abstain from Homalomena occuta.</p><p><b>METHODS</b>Extracts from the plant with 95% alcohol were distributed by several solvents and isolated via column chromatography on silica and Saphadex 20-LH gel.</p><p><b>RESULTS</b>Thirteen compounds were isolated from this plant. Among them seven natural products were identified via spectral methods as beta-stigmastol(H1) beta-D-Glucopyranoside(3)- stigmast-5-en-3-yl(H2); oplodiol(1); oplopanone(2); homalomenol(3); bullatantriol(4); 1 beta, 4 beta, 7 alpha-trihydroxyeudesmane(5).</p><p><b>CONCLUSION</b>All these compounds were isolated from this plant for the first time.</p>