ABSTRACT
OBJECTIVE：To establish a method for the simultaneous determination of euphol and euphorbol in Euphorbium resinifera，and to optimize the extraction technology. METHODS ：HPLC method was adopted. The determination was performed on Agilent ZORBAX Eclipse Plus C 8 column with mobile phase consisted of acetonitrile-water （90∶10，V/V）at the flow rate of 1.0 mL/min. The detection wavelength was set at 210 nm，and column temperature was 30 ℃. The sample size was 10 μL. Using the contents of euphol and euphorbol ，yield of the extract as evaluation index ，comprehensive score was conducted. The extraction technology was optimized by L （9 34）orthogonal tests ，with ethanol volume fraction ，extraction time and solvent dosage as factors. RESULTS：The linear ranges of euphol and euphorbol were 0.030 4-1.216 mg/mL（r＝0.999 6）and 0.01-0.4 mg/mL（r＝0.999 9）， respectively. RSDs of precision ，stability（24 h）and producibility tests were all lower than 2％. Average recoveries were 100.46％ （RSD＝1.03％，n＝6）and 99.36％（RSD＝0.91％，n＝6）. The optimized technology was extracting with 40 mL 95％ ethanol for 1 h. After 3 times of validation tests showed that average content of euphol was 94.43 mg/g（RSD＝0.92％，n＝3），and that of euphorbol was 25.42 mg/g（RSD＝0.98％，n＝3）；average yield of the extract was 51.42％（RSD＝1.95％，n＝3），and average comprehensive score was 98.87（RSD＝0.92％，n＝3）. CONCLUSIONS ：Established method is simple ，accurate and reproducible ， which can be used for the quality control of E. resinifera . The optimized extraction technology is simple and stable.
ABSTRACT
OBJECTIVE:To establish a preparative separation method for euphol reference substance in Euphorbia pekinensis. METHODS:Parts of petroleum ether extraction from E. pekinensis ethanol extract were separated by silica gel column chromatogra-phy with petroleum ether-ethyl acetate(95∶5-70∶30,V/V)by gradient elution. The enriched fractions of euphol were collected. With the methanol repeated recrystallization,nuclear magnetic resonance spectroscopy(NMR)method,mass spectrometry(MS)meth-od and other spectroscopic methods were used to identify the chemical structures,and thin layer chromatography (TLC),UV, HPLC-UV,HPLC-MS were combined to detect the mass fraction. RESULTS:The mass fraction of euphol reference substance sepa-rated from E. pekinensis was>99%. CONCLUSIONS:The reference substance of euphol acquired by this method meet the relative requirements of the chemical reference substance in the content dertermination of TCM new drug quality standard. It provides chemi-cal reference substance for the quality control of E. pekinensis and prescription preparations containing E. pekinensis and the basic research of effective substances.
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Objective: To study the isolation and identification of the chemical constituents in the root tubers of Euphorbia kansui and screen the cytotoxic activity of monomer compounds. Methods: The silica gel column chromatography and high performance liquid chromatography (HPLC) methodx were used. The structures were identified by the spectrum data analysis (MS, 1H-NMR, and 13C-NMR); The cytotoxic activity of monomer compounds was determined by MTT method. Results: Nine compounds were identified from the ethyl acetate part, respectively. They were kansuinin B (1), 3-O-(2, 3-dimethylbutanoyl)-13-O-dodecanoyl-ingenol (2), 5-O-benzoyl-20-deoxyingenol (3), euphol (4), isoscopletin (5), adenosine (6), hydroxytyrosol (7), 2', 4', 7-trihydroxy isoflavone (8), and emodin (9). Among them compound 1 is jatrophane-type diterpene, compounds 2 and 3 are the ingenane-type diterpenes, compound 4 is triterpene compound. Conclusion: Compounds 6-8 are isolated from this plant for the first time. Compounds 1 and 9 have the good inhibition on intestinal epithelial cells ICE-6.
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Objective: To study the chemical constituents of Ledum palustre. Methods: Compounds were isolated from n-hexane fraction in the ethanol extract from L. palustre by silica gel column chromatography and HPLC. The structures of the chemical constituents were identified by analysis on their spectral data. Results: Thirteen compounds were isolated from n-hexane fraction in the ethanol extract from L. palustre and identified as euphol acetate (1), lanosta-8, 24-dien-3-one (2), taraxerol (3), labda-8(17), 14-dien-13-ol (4), (24S)-stigmast-4-en-3-one (5), euphol (6), isopimaric acid (7), larixyl acetate (8), 28-hydroxyl-lupenone (9), β-sitosterol (10), isocalamendiol (11), erythrodiol (12), and uvaol (13). Conclusion: Compounds 2, 4-9, 11, and 12 are first obtained from the plants of Ledum L.
ABSTRACT
The anti-inflammatory properties of Euphol (EU) a diterpene, isolated from the rhizomes of the plant Euphorbia acaulis, were evaluated. This study was designed to examine the effects of EU on inflammation in a rat model of pleurisy compared with a steroidal anti-inflammatory drug (DEX, dexamethasone). The rat model of pleurisy was used to evaluate the effectiveness of EU on leukocyte migration, exudation volume, proinflammatory cytokines i.e. Tumor Necrosis factor α (TNFα), Interleukin-1 (IL-1β) and Interleukin-6 (IL-6) as well as on proinflammatory mediators PGE2, nitrite/nitrate levels, and LTB4. Pleurisy was induced by carrageenan (cg ), administered by intra pleural route (i.pl.) and the leukocyte infiltration and inflammatory parameters analyzed 4 h after pleurisy induction. EU (8mg/kg-1, p.o.) treated groups caused significant decrease in the infiltration of neutrophils and exudate volume . There was an observed marked reduction in TNFα, IL-1β and IL-6 levels. Flow cytometric analysis of intracellular TNFα by leukocytes present in pleural exudates showed a significant decrease relative to the total extracellular TNFα inhibition in the pleural fluid. The proinflammatory mediators PGE2, LTB4 release and the nitrite/nitrate levels were remarkably deregulated which are interrelated to TNFα and IL-1β levels.
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Objective To study the producing method for Euphorbia pekinensis promoted by endophytic fungi.Methods The tissue culture plantlets were innoculated with endophytic fungi,then it was transplanted to the pods and cultured for one year.The biomass and content of terpenoids(isoeuphpekinensin and euphol)were determined.Results The growth of the E.pekinensis was promoted by endophytic fungi E4(Fusarium sp.Ⅰ)and E5(Fusarium sp.Ⅱ).The fresh weight of the root increased 49.45% and 59.74% for E4 and E5 treated,respectively.The plants dry weight coefficient increased 2.99% and 6.48% for E4 and E5 treated,respectively.The contents of isoeuphpekinensin and euphol in the plants were increased for tissue cultured in both bottles and pods.For the plants cultured in pods for one year,the isoeuphpekinensin and euphol increased 92.79% and 40%,respectively treated by E4.The isoeuphpekinensin and euphol increassed 105.32% and 241.38%,respectively treated by E5.Conclusion The endophytic fungi E4 and E5 could promote the growth of E.pekinensis and accumulated terpenoids.
ABSTRACT
Objective To elucidate the constituents from the root of Euphorbia pekinensis and to look for new products with antitumor activity.Methods The chromatography on silica gel was used and the structures were identified by IR,NMR,and MS spectral analyses and their physicochemical properties were comparied.Results Eight compounds were isolated and identified as octadecanol(Ⅰ),octadecanyl-3-methoxy-4-hydroxybenzeneacrylate(Ⅱ),?-sitosterol(Ⅲ),triacontanoic acid(Ⅳ),2,2′-dimethoxy-3,3′-dihydroxy-5,5′-oxo-6,6′-biphenylformic anhydride(Ⅴ),euphol(Ⅵ),tirucallol(Ⅶ),and euphpekinensin(Ⅷ).Conclusion The compounds Ⅰ,Ⅳ,Ⅵ,and Ⅶ are isolated from the roots of E.pekinensis for the first time.