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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.
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Apocynum , China , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Medicine, Chinese TraditionalABSTRACT
OBJECTIVE: To establish a method for simultaneous determination of vitamin B1, dihydralazine sulfate, vitamin B6 and vitamin B5 in Compound Apocynum venetum tablets I.METHODS: HPLC method was used for content determination of 8 batches of samples from 2 enterprises. The determination was performed on Waters XBridge C18 column with mobile phase consisted of acetonitrile-0. 2％ phosphoric acid (gradient elution) at the flow rate of 0. 5 mL/min. The detection wavelengths were set at 285 nm (vitamin Bi, dihydralazine sulfate, vitamin B6) and 210 nm (vitamin B5). The column temperature was 30 ℃, and sample size was 10 μL. RESULTS: The linear ranges of vitamin Bi, dihydralazine sulfate, vitamin B6 and vitamin B5 were 41. 66-208. 3 μg/mL (r=0. 999 9), 127. 0-635. 0 μg/mL (r=0. 999 9), 40. 74-203. 7 μg/mL (r=0. 999 9) and 22. 72-113. 6 μg/mL (r= 0. 999 9), respectively. The limits of quantitation were 0. 267, 0. 274, 0. 062, 0. 124 μg/mL, respectively. The limits of detection were 0. 095, 0. 088, 0. 021, 0. 039 μg/mL, respectively. RSDs of precision, stability and reproducibility tests were all lower than 2. 0％ (n=6). The average recoveries were 98. 70％ (RSD=0. 69％, n=9),99. 74％ (RSD=1. 2％, n=9), 99. 89％ (RSD= 1. 0％, n=9), 99. 25％ (RSD=1. 2％, n=9), respectively. Among 8 batches of samples, the contents of vitamin B1, dihydralazine sulfate, vitamin B6 and vitamin B5 were 91. 44％-98. 74％, 36. 68％-95. 23％, 92. 42％-97. 90％, 91. 36％-99. 48％ of labelled amount, respectively. The contents of dihydralazine sulfate were significantly different and differed by 2. 6 times between the lowest and the highest threshold value. CONCLUSIONS: The method is accurate, reliable, simple and can be used for simultaneous determination of the above 4 components in Compound A. venetum tablets I.
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Apocynum venetum belongs to apocynaceae and is a perennial medicinal plant, its stem is an important textile raw materials. The projection of potential geographic distribution of A. venetum has an important significance for the protection and sustainable utilization of the plant. This study was conducted to determine the potential geographic distribution of A. venetum and to project how climate change would affect its geographic distribution. The projection geographic distribution of A. venetum under current bioclimatic conditions in northern China was simulated using MaxEnt software based on species presence data at 44 locations and 19 bioclimatic parameters. The future distributions of A. venetum were also projected in 2050 and 2070 under the climate change scenarios of RCP2.6 and RCP8.5 described in 5th Assessment Report of the Intergovernmental Panel on Climate Change (IPCC). The result showed that min air temperature of the coldest month, annual mean air temperature, precipitation of the coldest quarter and mean air temperature of the wettest quarter dominated the geographic distribution of A. venetum. Under current climate, the suitable habitats of A. venetum is 11.94% in China, the suitable habitats are mainly located in the middle of Xinjiang, in the northern part of Gansu, in the southern part of Neimeng, in the northern part of Ningxia, in the middle and northern part of Shaanxi, in the southern part of Shanxi, in the middle and northern part of Henan, in the middle and southern part of Hebei, Shandong, Tianjin, in the southern part of Liaoning and part of Beijing. From 2050 to 2070, the model outputs indicated that the suitable habitats of A. venetum would decrease under the climate change scenarios of RCP2.6 and RCP8.5.
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Objective To study the antidepressant effect and underlying mechanism of flavonoids extracted from Apocynum venetum leaves (AVL -extracts)in rat depression model.Methods The rats were randomly divided into seven parts including the control group,depression model group,low dose AVL-extracts prevention group (40 mg/kg),high dose AVL-extracts prevention group (80 mg/kg),low dose AVL-extracts treatment group (40 mg/kg),high dose AVL-extracts treatment group (80 mg/kg ) and fluoxetine treatment group, respectively. The rat depression model was established by the method of chronic unpredictable mild stress (CUMS ).The open field test was used to evaluate the autonomic activities.The relative expression levels of S1 00A1 0 mRNA were detected by Real-time PCR.Results The open field activities decreased significantly in the depression model group including horizontal movements and vertical movements with comparison to the normal group (P <0. 05 ).The autonomic activities increased significantly after preventive administration of high dose AVL-extracts (P<0. 05 ).The antidepressant effect of AVL-extracts was similar to that of fluoxetine in increasing the autonomic activities.The S1 00A1 0 mRNA expressed in the depressive group were higher than in the control group (P<0. 05 ),and the expression levels were significantly decreased after fluoxetine and AVL-extracts administration (P <0. 05 ).The high dose AVL -extracts administration group and fluoxetine group showed no significant difference in the expression levels of S1 00A1 0 mRNA when compared to the control group (P>0. 05 ),whereas the levels of those in low dose AVL-extracts administration groups were significantly lower than those in the control group (P<0. 05 ).Conclusion The study suggests that AVL -extracts have antidepressant effects on rat depression model.The underling mechanism of AVL-extracts antidepressant effect may be associated with regulation of S1 00A1 0 mRNA expression.
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Objective To investigate the anti-atherosclerosis effect of apocynum venetum(AV)by observing the influence of AV extract on early inflammatory factor TNF-αexpression.Methods Human U937 monocytes were differentiated to macrophages by the phorbol myristate acetate(PMA)induction and acted with 100 mg/L ox-LDL to form the foam cells for establishing the early atherosclerosis model(ox-LDL group).The different concentrations(0.2,0.4,0.8 mg/L)of AV were added to co-culture for 48 h (AV1,AV2,AV3 groups).The expression level of TNF-αin the supernate was detected by ELISA and RT-PCR respectively.Re-sults Compared with control group,the expression level of TNF-αin the ox-LDL group was significantly increased,the expression level of TNF-αin various AV medication groups(AV1,AV2,AV3 groups)was significantly decreased compared with the ox-LDL group(P<0.05).The AV concentration increase was negatively correlated with the TNF-αexpression level(P<0.05).Conclusion TNF-αis an important inflammatory factor in early atherosclerosis,AV could play the anti-atherosclerosis role by inhibiting the inflammatory factors.
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In this paper, the patents of traditional medicine A pocynum venetum were retrieved till 29th May, 2014, and the patent information, including the authorization condition, developing trends, geographic distribution, competi-tion agencies and therapeutic effects, was analyzed. Totally 764 patent families were obtained,only eight among them coming from countries and districts except China and Japan. China was in a leader position on the number of patent application of A . venetum. The patent applications mainly came from pharmaceutical enterprises. Most frequently patents were about innovations on the pharmacological effects such as regulation of blood pressure and blood sugar, as well as the development of health food and products. Traditional medicines used compatible included blood-activating and stasis-eliminating drugs, heat-clearing drugs, diaphoretics, deficiency-nourishing drugs and digestive drugs. This article revealed the trend and some rules of development of A . venetum, and reflected the developmental achievement and world patent pattern after a meta-analysis, exhorting to guide a better understanding and further research.
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Objective: To establish an HPLC method for the simultaneous determination of the main flavonoids in the leaves of Apocynum venetum and Poacynum hendersonii, and to reveal the differences of flavonoids in Apocyni Veneti Herba located between Xinjiang and other areas in China. Methods: The leaves of the two kinds of Apocyni Veneti Herba were extracted with 60% ethanol by reflux extraction. The separation was carried out on an Agilent TC-C18 column (150 mm × 4.6 mm, 5 μm) eluted with the mobile phases of acetonitrile-0.1% phosphoric acid (16:84). The column temperature was 40°C, and the flow rate was 1.0 mL/min, the detection wavelength was 360 nm, and the column temperature was 40°C. Results: The linear ranges of rutin, hyperin, and isoquercitrin were 4.96-49.60 (r = 0.9992), 0.96-9.60 (r = 0.9990) and 10.24-102.40 μg/mL (r = 0.9970), respectively; and the average recoveries (n = 5) were 98.2% (RSD = 1.5%), 92.3% (RSD = 3.2%), and 100.1% (RSD = 2.7%), respectively. Conclusion: This method is simple, rapid, accurate, and reproducible, and the results show that the flavonoids in Apocyni Veneti Herba located in Xinjiang are remarkably different from those in the other areas in China. In the leaves of A. venetum located in Xinjiang, the lutin content is high, about 0.55%, close to the isoquercetin content, but the hypercetin content is very low, only about 0.07%, and in the leaves of P. hendersonii, the contents of both lutin and hypercetin are very small. Such study will offer the credible method for the quality control of A. venetum and P. hendersonii.
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Objective: To study the nonflavonoid constituents in the leaves of Apocynum venetum, the medicinal halophyte. Methods: The constituents were isolated and purified by silica gel and Sephadex LH-20 column chromatography as well as preparative HPLC, and their structures were elucidated by means of physicochemical properties and spectrosocpic analyses. Results: Fifteen nonflavonoid compounds were isolated from the methanol extract from the leaves of A. venetum, and identified as scopoletin (1), esculetin (2), methyl chlorogenate (3), chlorogenic acid (4), grasshopper ketone (5), benzyl-O-β-D-glucopyranoside (6), 2-phenylethyl-O-β-D-glucopyranoside (7), 1-β-O-benzoyl-D-glucopyranoside (8), tyrosol (9), isovanillic acid (10), vanillic acid (11), protocatechuic acid (12), lupeol (13), β-amyrin (14), and α-linolenic acid (15). Conclusion: Compounds 2, 3, 5-10, 14, and 15 are isolated from this plant and the plants in Apocynum Linn. for the first time.
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Objective: To establish an optimized HPLC method for simultaneous determination of rutin, hyperoside, isoquercitrin, and quercetin in Apocynum venetum obtained from different locations. Methods: The separation was performed on a Shiseido C18 (3.0 μm, 30mmX100mm) column by gradient elution with acetonitrile: methanol = 10: 1 as the mobile phase A and 0.1% formic asid as the mobile phase B (0-4 min, 5% → 16%A; 4-15 min, 16% → 23%A; 15-20 min, 23% → 35%A; 20-22 min, 35% → 55%A) at a flow rate of 0.6 ml • min-1. The detection wave length of DAD was set at 360 nm, and the column temperature was 30°C. The injection volume was 5 μl. Results: Rutin, hyperoside, isoquercitrin, and quercetin were separated at base line within 22 min with good linearity (r = 0.9999), with the linearity range being 0.308 4 ~ 30.84 μg • ml-1 (r = 0.9999), 0.877 6-87.76 μg • ml-1 (r=0.9999), 0.9020 ~ 90.20 μg • ml-1(r = 0.9999),and 0.2498 ~ 24.98 μg • ml-1 (r = 0.9999), respectively. The result of intra-day and inter-day precisions, limits of detection and quantitation were all within the normal ranges. The recovery rates (n = 3) were 102.0%, 96.40%, and 103.8% for rutin, 98.50%, 101.0%, and 102.9% for hyperoside, 98.40%, 100.4%, and 101.4% for isoquercitrin, and 104.4%, 103.1%, and 103.5% for quercetin. The contents of the above 4 flavonoids were determined in Apocynum venetum from 9 different locations. Conclusion: The method developed in this study is rapid, simple, accurate, reliable, and with good repeatability; the method provides evidence for the quality control of Apocynum venetum.
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OBJECTIVE: To determine the contents of total flavonoids and quercetin in different parts of Apocynum venetum collected in Songnen grassland and study their antioxidant activities so as to provide references for efficient utilization of the herbal plant.METHODS: Ultrasonic extraction was applied to extract total flavonoids.The content of total flavonoids was determined by chromatometry and the content of quercetin was determined by HPLC.The antioxidant activities of the extracts from different parts of A.venetum were evaluated by determining their abilities in scavenging 1,1-diphenyl-2-picrylhydrazyl(DPPH?) radicals and?OH radicals.RESULTS: The contents of quercetin and total flavonoids were richer in both leaves and flowers of A.venetum than in stems.In addition,the total flavonoids extracted from the leaves and flowers of A.venetum showed higher antioxidant activity than from the stems.CONCLUSION: The leaves,flowers and stems of A.venetum can all be used as raw materials of natural antioxidants.
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OBJECTIVE:To establish the HPLC fingerprints of Apocynum Venetum leaves, determine the content of Quercetin in the sample and evaluate the variety and quality of commercial Apocynum Venetum leaves. METHODS: Quercetin was separated on Zorbarx-C18(250 mm?4.6 mm, 5 ?m),and the mobile phase consisted of methanol and 0.4% H3PO4(gradient elution) at a flow rate of 0.9 mL?min-1 with the detective wavelength set at 360 nm. RESULTS: The precision of Quercetin was satisfactory(RSD=0.21%,n=6),and the solution of Apocynum Venetum leaves was reproducible and stable within 10 hours. CONCLUSION: This study serves as a scientific basis for the identification and quality evalution of Apocynum Venetum leaves.