A method using high-performance liquid chromatography for the simultaneous determination of seven principal constituents including xanthoxylol, kakuol, methyleugenol,l-sesamin, safrole, sarisan, and l-asarinin in Asar...A method using high-performance liquid chromatography for the simultaneous determination of seven principal constituents including xanthoxylol, kakuol, methyleugenol,l-sesamin, safrole, sarisan, and l-asarinin in Asari Radix et Rhizoma (Xixin in Chinese) was developed. Fifty five samples of Asari Radix et Rhizoma from different sources (35 samples from Asarum heterotropoides var. mandshuricum, 3 samples from Asarum sieboldii var. seoulense, and 17 samples from Asarum sieboldii) were analyzed with this method. Chromatographic separation was achieved on an Agilent Zorbax SB-C18 column (4.6 mm×25 cm, 5 μm) using a gradient elution with mobile phase of aqueous (A) and acetonitrile (B). The assay was carried out at 20 ℃ and with detection at 287 nm. All calibration curves showed good linearity (r2〉0.999) within the tested ranges. The average recoveries were in the range of 96.0%-103.1% with relative standard deviation (RSD) less than 2.54%. The developed method was accurate with high sensitivity and good reproducibility. The content of volatile methyleugenol in LiaoXixin (the root and rhizome ofA. heterotropoides var. mandshuricum and A. sieboldii var. seoulense) was (6.17±5.13) rag/g, which was higher than (0.58±0.40) mg/g in HuaXixin (the root and rhizome ofA. sieboldii). The toxic safrole in LiaoXixin was (2.66±2.16) mg/g, which was lower than (6.44±3.89) mg/g in HuaXixin. This is the first report on the simultaneous and quantitative determination of anthoxylol and sarisan in Asari Radix et Rhizoma.展开更多
A rapid headspace single-drop micro-extraction(mix) gas chromatography mass spectrometry(SDMEGC -MS) for the analysis of the volatile compounds in Herba Asari was developed in this study.A mixed solvent of n-tride...A rapid headspace single-drop micro-extraction(mix) gas chromatography mass spectrometry(SDMEGC -MS) for the analysis of the volatile compounds in Herba Asari was developed in this study.A mixed solvent of n-tridecane and butyl acetate(1:1) was finally used for the extraction at 70 C for 15 min with sample amount of 0.750 g and 100 mesh particle size.Under the determined conditions,the pound samples of Herba Asari were directly applied for the analysis.SDME-GC-MS,SPME-GC-MS and SD-GCMS methods were compared and the results showed that SDME-GC-MS method was a simple, inexpensive and effective way to measure the volatile compounds in Herba Asari and could be used for the analysis of volatile compounds in complex samples.展开更多
OBJECTIVE: To investigate the effects of Ermiao Fang(EM) with medical guide Xixin(Herba Asari Mandshurici)(HAM) on bone marrow stem cell migration to a focal zone in osteoarthritis(OA) rats.METHODS: OA rats were induc...OBJECTIVE: To investigate the effects of Ermiao Fang(EM) with medical guide Xixin(Herba Asari Mandshurici)(HAM) on bone marrow stem cell migration to a focal zone in osteoarthritis(OA) rats.METHODS: OA rats were induced by arthrectomy and assigned to sham-operated, model, EM, or EM plus HAM groups.All rats were injected with recombinant human granulocyte colony-stimulating factor 30μg·kg-1·d-1for7 days and treated with EMor EM plus HAM at 1.6 or 1.9 g·kg-1·d-1 for 3 or 6 weeks, respectively. Chondrocyte apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining. Levels of interleukin-1 beta(IL-1β) tumor necrosis factor alpha(TNF-α) nitric oxide(NO), and inducible nitric oxide synthase(iNOS) in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay. Matrix metalloproteinases(MMPs)-13,tissue inhibitors of metalloproteinases(TIMPs)-1,Bromodeoxyuridine(BrdU), cluster of differentiation 34(CD34), and stromal cell-derived factor 1(SDF-1) were measured by immunohistochemical assay.RESULTS:The EM and EM plus HAM groups had significantly less cartilage damage and synovium inflammation the model group. Moreover, the EM and EM plus HAM groups had less chondrocyte apoptosis and more proteoglycan and collagen content than the model group.The EM and EMplus HAM groups had obviously higher MMPs-13 and TIMPs-1 expression in the cartilage than the model group. Moreover, the two formula groups had less release of IL-1β, TNF-α, NO, and iNOS than model group. Importantly, the expressions of BrdU, CD34,and SDF-1 in cartilage were significantly higher in the EM and EM plus HAM-Medtreated rats than model group. Notably, the EM plus HAM treatment seemed to have the greatest effects.CONCLUSION: HAM improves the therapeutic effects of EM on OA rats by enhancing BMSC directional homing to the focal zone.展开更多
基金National Basic Research Program of China (Grant No. 2006CB504707), National Science and Technology Major Projects for "Major New Drugs Innovation and Development" (Grant No. 2013ZX09508104) and National Natural Science Foundation of China (Grant No. 81274073).
文摘A method using high-performance liquid chromatography for the simultaneous determination of seven principal constituents including xanthoxylol, kakuol, methyleugenol,l-sesamin, safrole, sarisan, and l-asarinin in Asari Radix et Rhizoma (Xixin in Chinese) was developed. Fifty five samples of Asari Radix et Rhizoma from different sources (35 samples from Asarum heterotropoides var. mandshuricum, 3 samples from Asarum sieboldii var. seoulense, and 17 samples from Asarum sieboldii) were analyzed with this method. Chromatographic separation was achieved on an Agilent Zorbax SB-C18 column (4.6 mm×25 cm, 5 μm) using a gradient elution with mobile phase of aqueous (A) and acetonitrile (B). The assay was carried out at 20 ℃ and with detection at 287 nm. All calibration curves showed good linearity (r2〉0.999) within the tested ranges. The average recoveries were in the range of 96.0%-103.1% with relative standard deviation (RSD) less than 2.54%. The developed method was accurate with high sensitivity and good reproducibility. The content of volatile methyleugenol in LiaoXixin (the root and rhizome ofA. heterotropoides var. mandshuricum and A. sieboldii var. seoulense) was (6.17±5.13) rag/g, which was higher than (0.58±0.40) mg/g in HuaXixin (the root and rhizome ofA. sieboldii). The toxic safrole in LiaoXixin was (2.66±2.16) mg/g, which was lower than (6.44±3.89) mg/g in HuaXixin. This is the first report on the simultaneous and quantitative determination of anthoxylol and sarisan in Asari Radix et Rhizoma.
文摘A rapid headspace single-drop micro-extraction(mix) gas chromatography mass spectrometry(SDMEGC -MS) for the analysis of the volatile compounds in Herba Asari was developed in this study.A mixed solvent of n-tridecane and butyl acetate(1:1) was finally used for the extraction at 70 C for 15 min with sample amount of 0.750 g and 100 mesh particle size.Under the determined conditions,the pound samples of Herba Asari were directly applied for the analysis.SDME-GC-MS,SPME-GC-MS and SD-GCMS methods were compared and the results showed that SDME-GC-MS method was a simple, inexpensive and effective way to measure the volatile compounds in Herba Asari and could be used for the analysis of volatile compounds in complex samples.
基金Supported by Grants from the National Natural Science Foundation of China Project of Guiding Traditional Chinese Medicine Induced Bone Marrow Stem Cell Directional Homing to a Focal Zone for the Treatment of Osteoarthritis(No.81072900)
文摘OBJECTIVE: To investigate the effects of Ermiao Fang(EM) with medical guide Xixin(Herba Asari Mandshurici)(HAM) on bone marrow stem cell migration to a focal zone in osteoarthritis(OA) rats.METHODS: OA rats were induced by arthrectomy and assigned to sham-operated, model, EM, or EM plus HAM groups.All rats were injected with recombinant human granulocyte colony-stimulating factor 30μg·kg-1·d-1for7 days and treated with EMor EM plus HAM at 1.6 or 1.9 g·kg-1·d-1 for 3 or 6 weeks, respectively. Chondrocyte apoptosis and cartilage matrix components were tested by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling assay and special staining. Levels of interleukin-1 beta(IL-1β) tumor necrosis factor alpha(TNF-α) nitric oxide(NO), and inducible nitric oxide synthase(iNOS) in serum were detected by enzyme-linked immunosorbent assay or radioimmunoassay. Matrix metalloproteinases(MMPs)-13,tissue inhibitors of metalloproteinases(TIMPs)-1,Bromodeoxyuridine(BrdU), cluster of differentiation 34(CD34), and stromal cell-derived factor 1(SDF-1) were measured by immunohistochemical assay.RESULTS:The EM and EM plus HAM groups had significantly less cartilage damage and synovium inflammation the model group. Moreover, the EM and EM plus HAM groups had less chondrocyte apoptosis and more proteoglycan and collagen content than the model group.The EM and EMplus HAM groups had obviously higher MMPs-13 and TIMPs-1 expression in the cartilage than the model group. Moreover, the two formula groups had less release of IL-1β, TNF-α, NO, and iNOS than model group. Importantly, the expressions of BrdU, CD34,and SDF-1 in cartilage were significantly higher in the EM and EM plus HAM-Medtreated rats than model group. Notably, the EM plus HAM treatment seemed to have the greatest effects.CONCLUSION: HAM improves the therapeutic effects of EM on OA rats by enhancing BMSC directional homing to the focal zone.