In order to improve the quality standard of Marsdenia tenacissima, a quantitative determination method of tenacissoside H was developed using high performance liquid chromatography. The method was carried out on a YMC ODS-H80 (4.6 mm x 250 mm, 4 microm) column eluted with a mixture of acetonitrile and water (50:50) as the mobile phase. The flow rate was 0.8 mL x min(-1) and the column temperature was 35 degrees C. An evaporative light scattering detector (ELSD) was used with the temperature of drift tube set at 60 degrees C and the gas flow rate of nitrogen set at 1.5 mL x min(-1). The calibration curve was linear in the range from 0.5625 to 36.00 microg (r = 0.9998). The average recovery and RSD were 99.41% and 1.8%, respectively. The contents of tenacissoside H in the 11 samples from different habitats varied from 0.201% to 0.862%. The method established in this paper is specific and reliable to control and evaluate the quality of M. tenacissima.
Chromatography, High Pressure Liquid ; Glycosides ; analysis ; Marsdenia ; chemistry ; Reproducibility of Results

OBJECTIVETo establish a HPLC-ELSD fingerprint of Marsdenia tenacissima from different habitats, in order to provide a reliable method for scientific assessment and effective quality control.

METHODHPLC-ELSD was adopted to determine 25 baches of M. tenacissima herbs from different habitats. Traditional Chinese medicine chromatographic fingerprint similarity software assessment system 2004 developed by China Pharmacopoeia Committee was adopted to establish a common mode chart and assess chromatographic similarity based on the degree of correlation.

RESULTThe common mode for M. tenacissima herb C21 steroidal fingerprint was established, including 11 common characteristic peaks. Among them, 10 were identified. According to the assessment on the similarity of 25 batches of samples, 80% of them showed a similarity of over 0.80 in steroidal HPLC-ELSD fingerprint.

CONCLUSIONThe method can be used to assess the quality of M. tenacissima.

Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Ecosystem ; Light ; Marsdenia ; chemistry ; classification ; Scattering, Radiation

To ascertain current situation of wild Marsdenia tenacissima resources in Honghe, Yunnan province, the distribution, habitat characteristic and resources reserves of M. tenacissima were surveyed based on interviews and investigation. The results showed that M. tenacissima was found in 7 counties such as Jinping, Mengzi etc, and distributed mainly on the mountainsides from 800 m to 1 200 m. And distribution was affected by many factors, such as light, heat, topography, soil, and vegetation. M. tenacissima grew well in distribution areas. M. tenacissima had averagely a weight of 2.8 kg per plant. Resources reserve of M. tenacissima in Honghe was estimated to 1 300 tons by now but it reduced rapidly in resent years, the wild resources reserve may not meet demand of market. Resources protection and wildlife tending would be conducted to deal with increasing medication requirements.
China ; Ecosystem ; Marsdenia ; classification ; growth & development ; Plants, Medicinal ; classification ; growth & development ; Soil ; chemistry

OBJECTIVETo establish a quantitative method to deter mine C21 steroidal glycosides in Marsdenia tenacissima.

METHODMethanol was used as the extraction solvent and the samples were purified by macroporous resin ADS-7. A mixture of sulfuric acid-methanol (4:1)was used as color-producing reagent. The absorbance was measured at 325 nm.

RESULTThere is a good linearity (r = 0.999 9, n = 6) within the range of 10.6-148.4 microg. The average recoveries of tenacissoside-H at different concentrations were 95.8% to 97.1% with RSD less than 2.4% (n = 5).

CONCLUSIONThis method is reliable as a quantitative analytical method for the quality assessment of M. tenacissima.

Colorimetry ; methods ; Glycosides ; analysis ; Marsdenia ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Powders ; Quality Control ; Reproducibility of Results ; Steroids ; analysis

OBJECTIVETo identify the original plant of "Daibaijie", commonly used Dai herb.

METHODThe literature review, morphology and anatomy, pharmacognosy, molecular biology, chemistry were used to analysis.

RESULTDaibaijie's historical scientific name, Dregea sinensis Hemsl., was mistakenly given "Daibaijie" and D. sinensis have significant differences from the distribution, morphology and anatomy, pharmacognosy, molecular biology and chemical composition. "Daibaijie" matches with the characteristics of Marsdenia tenacissima (Roxb.) Moon in Flora of China in English.

CONCLUSIONDaibaijie's original plant is M. tenacissima (Roxb.) Moon. The description and illustration of M. tenacissima (Roxb.) Moon in Flora of China in China are wrong. The illustration of M. tenacissima in Flora of China in English is wrong too.

China ; ethnology ; Herbal Medicine ; Marsdenia ; anatomy & histology ; classification ; Medicine, Chinese Traditional ; Plant Components, Aerial ; anatomy & histology ; classification

AIMTo study the chemical constituents of Marsdenia tenacissima (Roxb.) Wight et Arn.

METHODSTo separate compounds with various chromatography technology and to elucidate their structures by chemical and spectral analysis.

RESULTSTwo compounds were isolated from Marsdenia tenacissima and their stuctures were determined as tenacissosides J (I) and tenacissosides K (II).

CONCLUSIONCompounds I and II are new C21 steroidal glycosides.

Glycosides ; chemistry ; isolation & purification ; Marsdenia ; chemistry ; Molecular Conformation ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Steroids ; chemistry ; isolation & purification

To study the C21 steroids of the stems of Marsdenia tenacissima (Roxb.) Wight et Arn, various chromatography methods were used for the isolation of the constituents and their structures were identified by spectral analysis. Eight C21 steroids were isolated from the CHCl3 extract, which were identified as 11alpha-O-tigloyl-17beta-tenacigenin B (1), 17beta-tenacigenin B (2), tenacigenoside A (3), 11alpha-O-2-methylbutyryl-12beta-O-acetyl tenacigenin B (4), tenacissoside H (5), marsdenoside A (6), tenacissoside G (7), and tenacissoside I (8). Among them, compound 1 is a new compound.
Marsdenia ; chemistry ; Molecular Structure ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Saponins ; chemistry ; isolation & purification ; Steroids ; chemistry ; isolation & purification

OBJECTIVETo study characteristic fingerprints of the fat-soluble components of different Marsdenia tenacissima samples by GC and GC-MS.

METHODThe sample was split in the 280 degrees C injection port, with 10:1 split ratio, and separated on a fused silica capillary column (DB-5, 30 m x 0.25 mm I. D., 0.25 microm film). The temperature program was 70 degrees C for 5 min, 8 degrees C x min(-1) to 200 degrees C, then 5 degrees C x min-1 to 260 degrees C, 260 degrees C for 10 min. The high pure nitrogen was used as a carrier gas, The FID temperature was 300 degrees C.

RESULTThere are 15 common peaks in the GC fingerprints of M. tenacissima and they are identified by GC-MS. The analyses on cluster and similarity degree of the finger the crude drugs from various habitats have been performed.

CONCLUSIONThe method has good repeatability. The GC fingerprint combined with the HPLC fingerprint of M. tenacissim can be used to evaluate its quality integrally.

China ; Chromatography, Gas ; methods ; Drugs, Chinese Herbal ; analysis ; chemistry ; standards ; Ecosystem ; Gas Chromatography-Mass Spectrometry ; Marsdenia ; chemistry ; growth & development ; Plants, Medicinal ; chemistry ; growth & development ; Quality Control ; Reproducibility of Results

OBJECTIVEMethylation-specific epigenetic process and gene expression profiles of HeLa cells treated with ultra-high dilutions (HDs) of two plant extracts, Hydrastis canadensis (HC-30) and Marsdenia condurango (Condu-30), diluted 1060 times, were analyzed against placebo 30C (Pl-30) for alterations in gene profiles linked to epigenetic modifications.

METHODSSeparate groups of cells were subjected to treatment of Condu-30, HC-30, and Pl-30 prepared by serial dilutions and succussions. Global microarray data recorded on Affymetrix platform, using 25-mer probes were provided by iLifeDiscoveries, India. Slides were scanned with 3000 7G microarray scanner and raw data sets were extracted from Cel (raw intensity) files. Analyses of global microarray data profile, differential gene expression, fold change and clusters were made using GeneSpring GX12.5 software and standard normalization procedure. Before microarray study, concentration of RNA (ng/μL), RIN value and rRNA ratio for all the samples were analysed by Agilant Bioanalyzer 2100. Reverse transcriptase polymerase chain reaction (RT-PCR) and quantitative RT-PCR were done for analyzing SMAD-4 expression. Fluorescence-activated cell sorting study was further made to elucidate fate of cells at divisional stages. Methylation-specific restriction enzyme assay was conducted for ascertaining methylation status of DNA at specific sites.

RESULTSHDs of HC-30 and Condu-30 differentially altered methylation in specific regions of DNA and expression profiles of certain genes linked to carcinogenesis, as compared to Pl-30. Two separate cut sites were found in genomic DNA of untreated and placebo-treated HeLa cells when digested with McrBC, compared to a single cut observed in Condu-30-treated genomic DNA. SMAD-4 gene expression validated the expression pattern observed in microarray profile. Methylation-specific restriction enzyme assay elucidated differential epigenetic modifications in drug-treated and control cells.

CONCLUSIONHDs triggered epigenetic modifications and alterations in microarray gene expression profiles of many genes associated with carcinogenesis in HeLa cells in vitro.

Cell Cycle ; Cluster Analysis ; DNA Methylation ; Epigenesis, Genetic ; drug effects ; HeLa Cells ; Humans ; Hydrastis ; Marsdenia ; Plant Extracts ; pharmacology ; Transcriptome ; drug effects

In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; classification ; standards ; Geography ; Marsdenia ; chemistry ; classification ; Medicine, Chinese Traditional ; Principal Component Analysis ; Quality Control ; Reproducibility of Results ; Spectroscopy, Fourier Transform Infrared ; methods