This study established high-performance liquid chromatography(HPLC) fingerprints of Chinese medicines derived from Apocynum venetum and Poacynum pictum in Xinjiang and explored their composition differences with the combination of content determination, similarity analysis, cluster analysis and principal component analysis. The HPLC conditions included Phenomenex Kinetex C_(18) column(4.6 mm ×100 mm, 2.6 μm), acetonitrile-0.01% trifluoroacetic acid aqueous solution as mobile phase, gradient elution, flow rate of 0.6 mL·min~(-1), detection wavelength of 281 nm and column temperature of 25 ℃. The content of chlorogenic acid, quercetin-3-O-sophoroside, rutin, hyperin, isoquercitrin, trifolin and astragalin was determined in 31 batches of medicinal materials, and fingerprint research and chemometric analysis were performed with Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(Version 2004 A) and SPSS 21.0. In the Chinese Pharmacopoeia 2020, the quality of Apocyni Veneti Folium is controlled by character identification, microscopic identification, thin layer chromatography identification and quantitative determination of hyperin. There were 21 common peaks of A. venetum and P. pictum in the HPLC fingerprints, 5 of which were identified as chlorogenic acid, hyperin, isoquercitrin, trifolin and astragalin, with their content also determined. Except for 3 batches of medicinal materials, the similarity of other 28 batches was higher than 0.83, indicating good similarity. Two categories were formed in the cluster analysis based on content determination, which showed that some differences existed in similarities between different regions of Xinjiang. The medicinal materials were ranked by quality with principal component analysis, and the results indicated that the top 15 all came from northern Xinjiang. The quality difference of A. venetum and P. pictum had a correlation with the place of origin. This study provides a reference for the analysis and evaluation of A. venetum and P. pictum from different habitats and the selection of introduction and cultivation areas.
Apocynum ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; Medicine, Chinese Traditional

OBJECTIVETo establish a sensitive and specific HPLC fingerprint for the quality controlling of total flavonoids of Folium Apocyni Veneti.

METHODHPlC analysis was performed on a Kromasil C18 column (4.6 mm x 250 mm, 5 microm) with the mixture of solvent A [acetonitrile-phosphoric acid (95:5)] and solvent B (0.05% phosphoric acid) in gradient mode at a flow rate of 1.0 mL x min(-1). The detection wavelength was set at 360 nm. The column temperature was set at 25 degrees C and the injection volume was 20 microL.

RESULTThe chromatographic fingerprint of total flavonoids was established which showed 17 characteristic peaks from 7 patches of total flavonoids products. The similarity from different patches was 0.95-1.00 analyzed by the software of 'Computer-aided Similarity Evaluation' and showed high similitude in peak numbers and the retention time. Moreover, comparison of the HPLC profiles of the total flavonoids with the corresponding Folium Apocyni Veneti leaves indicated that they were closely related to each other.

CONCLUSIONThe chromatographic fingerprint of the total flavonoids with high specificity and can be used to control its quality and assure the homogenicity for each patch of the total flavonoids.

Apocynum ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; chemistry ; Reproducibility of Results

OBJECTIVETo develop a high-performance liquid chromatography (HPLC) method for the simultaneous determination of rutin, hyperoside, isoquercetin, astragulin, quercetin, and kaempferol in Apocynum venetum and its extracts.

METHODThe separation was carried out on a Shim pack ODS (4.6 mm x 250 mm, 5 microm) colum eluted with in mobile phases of water containing 0.2% phosphoric acid and acetonitrile containing 0.2% phosphoric acid in acetonitrile gradient mode. The column temperature was 40 degrees C, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm.

RESULTThe good seperation of six flavonoids was achieved within 40 min, with the relative standard deviations (RSD) of intra- and inter-day precision < or = 2.0%. Calibration curves of rutin, hyperoside, isoquercetin, astragulin, quercetin, and kaempferol showed good linear relationship (R2 > 0.999 7, n = 6). The average recoveries of the six flavonoids were within 97.30% - 105.8% (RSD 2.6%). Three batches of A. venetum and 2 batches of its extracts were determined.

CONCLUSIONThe developed method is simple, accurate, and repeatable, and can be readily used as a powerful tool for the quality control of A. venetum and its extracts.

Apocynum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; Quality Control ; Reproducibility of Results

In this study, laser scanning confocal microscopy (LSCM) was used to determine the location and relative quantity of flavonoids in the leaves of Apocynum venetum L. from the top, middle and basal parts of the branch. The leaves of the plants of one, two and three years old, separately, were collected in July. ANOVA and LSD test were employed in the statistical analysis. The results indicated that flavonoids located mainly in xylem conduit of vein, collenchyma, epidermic cells and cuticle. The data of flavonoids contents under statistical analysis showed that difference existed in the leaves of different parts and different ages. This study provided the reliable scientific material about the analysis of the ecological and the exploitation of the leaves of Apocynum venetom L.
Apocynum ; chemistry ; Flavonoids ; analysis ; Microscopy, Confocal ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Time Factors

A system for the Agrobacterium rhizogenes-mediated transformation and plant regeneration of A. venetum has been developed. The highest transformation frequency was 100%, achieved by using strain LBA9402 with root explants. The highest density of hairy roots reached 22 when root explants transformed by R1000 cultured in the dark. Adventitious shoots were obtained from profusely branched, fast-growing (type PBF) hairy roots, and the adventitious shoot induction frequency was 20%. Regenerated shoots rooted easily on hormone-free 1/2 MS solid medium in 2 weeks. Approximately 1/3 regenerated plants derived from hairy roots exhibited prolific roots with shortened internodes. Whereas other regenerated plants showed another phenotype: long intemodes, strong stems, and fleshy blades. However, all regenerated plants displayed a relatively fast development procedure and stronger than the aseptic seedlings. Polymerase chain reaction (PCR) analyses confirmed the hairy root lines and regenerated plants were induced by A. rhizogenes.
Apocynum ; genetics ; growth & development ; Plant Roots ; growth & development ; Regeneration ; Rhizobium ; genetics ; Tissue Culture Techniques ; Transformation, Genetic

OBJECTIVETo study the chemical constituents of the flowers of Apocynum venetum.

METHODChromatographic methods were used to isolate compounds from the flowers of A. venetum and spectral methods were used to identify the structures of the isolated compounds.

RESULTSeven compounds, kaempferol (I), quercetin (II), quercetin-3-O-beta-D-glucoside (III), kaempferol-3-O-beta-D-glucoside (IV), vanillic acid (V), baimaside (VI), daucosterol (III), were isolated from the flowers of A. venetum.

CONCLUSIONCompound I, V, VI, VII were obtained from this plant for the first time.

Apocynum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Flowers ; chemistry ; Kaempferols ; isolation & purification ; Sitosterols ; isolation & purification ; Vanillic Acid ; isolation & purification

OBJECTIVETo study the flavonoids in the leaves of Apocynum ventum.

METHODCompounds were separated by repeated polyacrylamide column chromatgraphy combined with Sephadex LH-20 column chromatgraphy, their structures were confirmed by the analysis of spectral data and physicochemical properties.

RESULTSix compounds were isolated from A. ventum. They were elucidated as quercetin (1), kaempferol (2), kaempferol-3-O-(6"-O-acetyl)-beta-D-glucopyranoside (3), quercetin-3-O-(6"-O-acetyl )-beta-D-glucopyranoside (4), quercetin-3-O-(6"-O-acetyl)-beta-D-galactopyranoside (5), kaempferol-7-O-alpha-L-rhamnopyranoside (6).

CONCLUSIONCompound 5 and 6 were isolated from the genus Apocynum for the first time.

Apocynum ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Kaempferols ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification

OBJECTIVETo investigate the purification process of total flavones and total tannins from Apocynum venetum leaves with macroreticular resin.

METHODThe static capacity absorption, dynamic absorption ratio and dynamic elution ratio of different types of macroreticular resins were studied and compared in order to find the best one among the eight macroreticular resins, and the technical process of the type of HPD-400 type macroreticular resin was studied in detail.

RESULTThe type of HPD-400 type macroreticular resin showed the best comprehensive absorption property with the following technological conditions: the current velocity was 1 BV x h(-1), the volume of distilled water was 2 BV, the eluting reagent was 60% ethanol, and the volume of 60% ethanol was 3 BV.

CONCLUSIONThe purity of total flavones and total tannins in the product is up to 80% after purified with HPD-400 macroreticular resin. Therefore, this purification process is feasible.

Apocynum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavones ; analysis ; Plants, Edible ; chemistry ; Plants, Medicinal ; chemistry ; Reference Books, Medical ; Tannins ; analysis

OBJECTIVETo investigate the purification process of total flavones and total tannins from Apocynum venetum leaves with macroreticular resin.

METHODThe static capacity absorption, dynamic absorption ratio and dynamic elution ratio of different types of macroreticular resins were studied and compared in order to find the best one among the eight macroreticular resins, and the technical process of the type of HPD-400 type macroreticular resin was studied in detail.

RESULTThe type of HPD-400 type macroreticular resin showed the best comprehensive absorption property with the following technological conditions: the current velocity was 1 BV x h(-1), the volume of distilled water was 2 BV, the eluting reagent was 60% ethanol, and the volume of 60% ethanol was 3 BV.

CONCLUSIONThe purity of total flavones and total tannins in the product is up to 80% after purified with HPD-400 macroreticular resin. Therefore, this purification process is feasible.

Adsorption ; Apocynum ; chemistry ; Chromatography, Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavones ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Resins, Synthetic ; chemistry ; Tannins ; chemistry ; isolation & purification

To investigate the resources of medicinal plant, such as wild Apocynum, supervised classification based on Principal Component Analysis (PCA) and texture feature were used to monitor wild medicinal plants from image captured by ZY-3 and World-view-2 and compare which satellite Image are more appropriate to monitor the wild medicinal plants. The research results shows that: for more complex growth conditions wild medicinal plants Apocynum, high-resolution images Worldview-2 is more suitable for its remote identification, the low-resolution satellite ZY-3 can only recognizes the wild medicinal plants which distributed intensively. If the study target distribution is more intensive and larger scale, and cultivated type medicinal plants, the use of satellite ZY-3 in low resolution remote sensing data to identify the target can be a good choice, it is not necessary to buy high-resolution data, in order to avoid waste of expenditure, for the scattered distribution, the high-resolution satellite imagery data may be indispensable to identify targets.
Apocynum ; chemistry ; growth & development ; China ; Conservation of Natural Resources ; Geographic Information Systems ; Plant Dispersal ; Plants, Medicinal ; chemistry ; growth & development ; Remote Sensing Technology ; methods