Rapid Identification of Alkaloids in Evodia rutaecarpa by HPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

Objective:To obtain the information of alkaloids in Evodia rutaecarpa by HPLC-Q-TOF-MS/MS. Method:Inter Sustain-C18 column (4.6 mm×250 mm,5 μm) was used with 0.2% formic acid water-acetonitrile as the mobile phase for gradient elution. The column temperature was 25℃,the volume flow rate was 1.0 mL·min-1,and the sample volume is 5 μL. The detection wavelength was 245 nm,and the chromatographic effluent was detected and analyzed by using both positive and negative ions. Result:According to molecular ion peaks and secondary mass spectrometry characteristic fragment ions,as well as the mass spectrometry information of reference substances and relevant literature reports,more than 40 major peaks were analyzed,and 21 alkaloids were identified from the methanol extract of E. rutaecarpa, including 10 kinds of indole alkaloids,10 kinds of quinolone alkaloids,and 1 kind of ephedrine. Main types of alkaloids in E. rutaecarpa were basically clarified. And the research found that the alkaloids have a good response mainly in the positive mode. Conclusion:Based on HPLC-Q-TOF-MS/MS technology, high-performance liquid chromatography (HPLC) separation,mass spectrometry determination of molecular mass,pyrolysis data,literature analysis and retrieval were performed to quickly,accurately and comprehensively identify alkaloids in E. rutaecarpa, so as to provide a scientific basis for the further extraction and separation of the chemical constituents of E. rutaecarpa.

Evodiamine Reduces Caffeine-Induced Sleep Disturbances and Excitation in Mice

Worldwide, caffeine is among the most commonly used stimulatory substances. Unfortunately, significant caffeine consumption is associated with several adverse effects, ranging from sleep disturbances (including insomnia) to cardiovascular problems. This study investigates whether treatment with the Evodia rutaecarpa aqueous extract (ERAE) from berries and its major molecular component, evodiamine, can reduce the adverse caffeine-induced sleep-related and excitation effects. We combined measurements from the pentobarbital-induced sleep test, the open field test, and the locomotor activity test in mice that had been dosed with caffeine. We found that ERAE and evodiamine administration reduced the degree of caffeine-induced sleep disruption during the sleep test. Additionally, we found that evodiamine significantly inhibits caffeine-induced excitation during the open field test, as well as decreasing hyperlocomotion in the locomotor activity test. Additional in vitro experiments showed that caffeine administration decreased the expression of γ-aminobutyric acid (GABA)(A) receptor subunits in the mouse hypothalamus. However, evodiamine treatment significantly reversed this expression reduction. Taken together, our results demonstrate that ERAE and its major compound, evodiamine, provide an excellent candidate for the treatment or prevention of caffeine-induced sleep disturbances and excitatory states, and that the mechanism of these beneficial effects acts, at least in part, through the GABA(A)-ergic system.

Chemical constituents from the fruits of Evodia rutaecarpa and their biological activities / 中成药

AIM To study the chemical constituents from the fruits of Evodia rutaecarpa (Juss.) Benth.and their biological activities.METHODS The ethyl acetate fraction of 80％ ethanol extract from E.rutaecarpa was isolated and purified by silica column and recrystallization,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antitumor and antifungal activities were determined by MTT and mycelium growth method,respectively.RESULTS Ten compounds were isolated and identified as rutaecarpine (1),evodiamine (2),1-methyl-2-undecyl-4 (1H)-quinolone (3),1-methyl-2-tridecyl-4 (1H)-quinolone (4),wuchuyuamide Ⅰ (5),wuchuyuamide Ⅲ (6),wuchuyuamide Ⅳ (7),β-sitosterol (8),β-daucosterol (9),dehydroevodiamine (10).Compounds 1,2,10 had certain inhibitory effects on MDA-MB-231,LoVo,A2780 and HeLa with the IC50values of 0.65-29.45 μmol/L.Compound 1 had a certain inhibitory effect on Botrytis cinerea,Glomerella cingulata and Rhizoctonia solani with the inhibitory rates of 18.12％-49.57％.CONCLUSION Compounds 3-7 are isolated from this plant for the first time,compounds 1,2,10 have strong biological activities.

Simultaneous Determination of Chlorogenic Acid,Evodiamine and Rutecarpine in Different Places of Evodia rutaecarpa by UPLC / 中国药房

OBJECTIVE:To establish a method for the simultaneous determination of chlorogenic acid,evodiamine,rutecar-pine in different places of Evodia rutaecarpa. METHODS:UPLC was performed on the column of ACQUITY UPLC BEH C18 with mobile phase of acetonitrile-0.1% Phosphoric acid aqueous solution(gradient elution)at a flow rate of 0.40 ml/min,the detection wavelength was 326 nm and 220 nm,column temperature was 30 ℃,and the volume injection was 2 μl. RESULTS:The linear range was 7.67-76.67 μg/ml for chlorogenic acid(r=0.999 2),13.33-133.33 μg/ml for evodiamine(r=0.999 7)and 13.33-133.33μg/ml for rutecarpine(r=0.999 8);the limits of quantitation were 0.11 ng,0.05 ng and 0.05 ng,the limits of detection were 0.03 ng,0.01 ng and 0.01 ng,respectively;RSDs of precision,stability and reproducibility tests were lower than 3%;recoveries were 96.19%-101.90%(RSD=2.19%,n=6),95.35%-101.16%(RSD=2.27%,n=6)and 95.92%-98.98%(RSD=1.33%,n=6),re-spectively. CONCLUSIONS:The method is rapid and accurate,and suitable for the simultaneous determination of chlorogenic ac-id,evodiamine,rutecarpine in different places of E. rutaecarpa.

Effects of Four Interior-warming Drugs on the Tension of Ileum Smooth Muscle and Ca2+-ATPase in Rabbits / 医药导报

Objective To investigate the effects of four interior-warming drugs( galangal,cinnamon,evodia rutaecarpa,and dried ginger)on the tension of ileum smooth muscle and Ca2+-ATPase on the cell membrane in rabbits. Methods The effects of galangal,cinnamon,evodia rutaecarpa,and dried ginger were examined on normal ileum smooth muscle,in vitro intestinal muscle contraction caused by acetylcholine(ACh),barium chloride(BaCl2 )and histamine(His), and ACh-induced calcium release by using BL-420E+ biological signal collection and processing system.The average tension was measured within 1 min before delivery and within 3 minutes after the treatment,and the inhibition rate was calculated according to the average tension value.The effects of sera containing galangal,cinnamon,evodia rutaecarpa,and dried ginger on Ca2+-ATPase activity on the cell membrane of the intestinal smooth muscle were examined by phosphorus method. Results Galangal,cinnamon,evodia rutaecarpa,and dried ginger at high concentrations could restrain in vitro intestinal contraction in normal circumstances(P<0.05 or P<0.01).Significant inhibitory effects on intestinal contraction caused by ACh,His and BaCl2 were found in low,medium and high concentration groups(P<0.01).There was a dose-effectiveness relationship between the inhibition rate and final drug concentrations.The ACh-induced intracellular and extracellular calcium dependent contraction were significantly inhibited by the four interior-warming drugs( P < 0. 05 or P < 0. 01). The Ca2+-ATPase activities were( 0. 384 ± 0.070),(0.302±0.016),(0.307±0.016),(0.296±0.016),(0.313±0.003)U·mg-1 ,respectively,in intestinal smooth muscle in normal control group and high concentration groups of galangal,cinnamon,evodia rutaecarpa,and dried ginger(P<0.01). Conclusion Interior-warming drugs may relax intestinal smooth muscle by reducing the intracellular calcium release and the extracellular calcium inflow via receptor-controlled calcium channels,and inhibiting the Ca2+-ATPase activity in smooth muscle.

Separation of Five Quinolone Alkaloids from Fruits of Evodia rutaecarpa by High-speed Counter-current Chromatography / 中草药·英文版

Objective: To develop a suitable method for the large-scale separation of quinolone alkaloids from the fruits of Evodia rutaecarpa by high-speed counter-current chromatography (HSCCC). Methods: Two solvent systems were developed for the separation method. The upper phase was used as the stationary phase, and the lower phase was used as the mobile phase at 35 °C with the flow rate of 2 mL/min and rotation speed of 855 r/min. Results Using the described method, 1-methyl-2-undecyl-4(1H)-quinolone (1), evocarpine (2), 1-methy-2-[(6Z,9Z)]-6,9-pentade-cadienyl-4-(1H)-quinolone (3), dihydroevocarpine (4), and the mixture (5) of 1-methyl-2-[(Z)-10-pentadecenyl]-4(1H)-quinolone (Va) and 1-methyl-2-[(Z)-6-pentadecenyl]-4(1H)-quinolone (Vb) could be isolated from a petroleum ether extract. They were identified by 1H-NMR, 13 C-NMR, and MS/MS, and the purities were 94.3%, 95.2%, 96.8%, 98.3%, and 96.8%, respectively. Conclusion: Five quinolone alkaloids from the fruits of E. rutaecarpa could be systematically isolated and purified using HSCCC. The presented method is simple and efficient with good potentials on the preparation of reference substances, especially on the quality control of Chinese materia medica. © 2014 Tianjin Press of Chinese Herbal Medicines.

Separation and purification of total alkaloids from Evodia rutaecarpa using 732 cation exchange resin and investigations on its anti-breast cancer bioactivity / 中国生化药物杂志

Objective To optimize conditions for extraction of alkaloids from Evodia rutaecarpa by means of 732 cation exchange resin and performe its anti-breast cancer bioactivity.Methods The optimum processing route on extraction of alkaloids from Evodia rutaecarpa was investigated by means of 732 cation exchange resin.The performance of 732 cation exchange resin was compared with tranditonal process ( aqueous extraction-ethanol precipitation in extraction-purification process).The alkaloid reagent-potassium mercuric iodide test and MTT test were performed on the crudes.Results Compared with traditional purification process, it was much better to use 732 cation exchange resin approach for extraction of alkaloids from Evodia rutaecarp with high yield(16.81%) The best eluent should be saturated brine with excellent purification.No obvious correlations were found between the toxin of breast cancer cell and concentration of total alkaloids.When the concentration of total alkaloids was 50μmol/mL, cellular survival rate was 68%afterward 24 h.When the concentration of total alkaloids comes to 100 μmol/mL and 150 μmol/mL, cellular survival rates were slightly decreased by 66% and 60%.Conclusion 732 cation exchange resin performes much higher than traditional process in purification of total alkaloids extracted from Evodia rutaecarpa.Simultaneously, the extracted total alkaloids showe remarkable inhibition in breast cancer cell.

Simultaneous Determination of Five Active Chemical Components in Tibetan Medicine Pterocephalus Hookeri Hoeck by UFLC-PDA / 世界科学技术-中医药现代化

This study was aimed to establish an Ultra Fast Liquid Chromatography-Photo Diode Array (UFLC-PDA) method for the simultaneous determination of five chemical components, which included chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, in Pterocephalus hookeri h eck. Agilent Poroshell 120 SB-C18 (100 mm í 4.6 mm, 2.7 μm) was adopted, with acetonitrile-0.2% phosphoric acid solution in gradient elution as the mobile phase at the flow rate of 1.0 mL·min-1. And the injection volume was 0.4 μL. The detection wavelength was set up at 237 nm and 325 nm. And the column temperature was 30℃. The results showed that the calibration curve was linear within the range of 8.72~218.0, 1.52~38.0, 2.44~61.0, 29.36~734.0, 3.00~75.0μg·mL-1 (r > 0.999 6, n=9) for chlorogenic acid, loganin, sweroside, evodia rutaecarpa glycosides and triplostoside A, respectively. The average recovery rates were 99.46%, 99.41%, 100.14%, 98.89%, and 99.42%, respectively. The RSD was 0.69%, 0.66%, 0.60%, 1.21%, and 0.64%, respectively (n = 9). It was concluded that this method was simple, accurate and reproducible, which can be used for the simultaneous determination of the content of five chemical components in P. hookeri.

Analgesic effects of Evodia extract cream and its mechanisms in mice / 中国药学杂志

OBJECTIVE: To study the analgesic effects of Evodia extract cream and its mechanisms. METHODS: Kunming mice were used as model animals, and hot plate test and formalin test were adopted in analgesia experiments. ELISA test was used to detect the contents of CGRP and PGE2. In addition, the tissue section technique was used to investigate the extent of inflammation cell infiltration in the area of pain. RESULTS: Evodia extract cream with a drug loading ranging from 0.1% to 0.4% could relieve pain induced by noxious heat (P < 0.05), and demonstrated significant analgesic effects in the mice formalin test (P < 0.05) in a dose-dependent manner. RESULTS: from ELISA test showed that Evodia extract cream with a drug loading of 0.2% significantly decreased the expression of CGRP in the pain zone of skin (P < 0.05) 3 min after subcutaneous injection of formalin (0.2% EE), and the expression of CGRP showed significant reduction over time (P < 0.05). But Evodia extract cream with a drug loading ranging from 0.1% to 0.4% had no significant effects on the contents of CGRP and PGE2 in plasma or the content of PGE2 in mice skin. Histological section RESULTS: indicated that the content of inflammation cell infiltration induced by formalin was decreased significantly after topical administration of Evodia extract cream with a drug loading ranging from 0.1% to 0.4% (P < 0.05). CONCLUSION: Topical administration of Evodia extract cream has significant dose-dependent analgesic effects. These findings suggest that the analgesic mechanism of Evodia extract cream is most likely related with exhausting the CGRP and interdicting its conduction and coordinating with anti-inflammatory effect.

Effect of Evodia rutaecarpa cultivated in Guizhou Yuqing on rabbit thoracic aorta in vitro and its mechanism / 中国药学杂志

OBJECTIVE: To investigate the effect of Evodia rutaecarpa cultivated in Guizhou Yuqing on rabbit thoracic aorta in vitro and its possible mechanism. METHODS: Evodia rutaecarpa water decoction at concentrations from 0.1 to 1.6 mg · mL-1 was cumulatively added into organ bath, the isometric tensions of rabbit thoracic aortic rings with intact endothelium or denuded endothelium were recorded, then the thoracic aortic rings were pre-incubated with L-NAME, indometacin and barium chloride respectively to observe the effect of Evodia rutaecarpa. RESULTS: The water decoction of Evodia rutaecarpa cultivated in Guizhou Yuqing could dose-dependently increase the tension of rabbit thoracic aortic rings, and the constrictive effect was stronger on endothelium-intact aortic rings than on endothelium-denuded aortic rings. The contraction induced by Evodia rutaecarpa on endothelium-intact aortic rings was decreased by pre-incubation with L-NAME and indometacin. However, Evodia rutaecarpa had no significant effect on endothelium-intact aortic rings when pre-incubated with barium chloride. CONCLUSION: The water decoction of Evodia rutaecarpa cultivated in Guizhou Yuqing has endothelium-dependent contracting effect on rabbit thoracic aorta, and it may also stimulate the release of NO and PGI2from endothelium cells, which partly compromises its vasoconstrictive effects.

Hepatoprotective effect of Evodia officinalis Dode on carbon tetrachloride induced liver injury in rats / 中国医师杂志

Objective To investigate the protective effect of ethanol extract from Evodia officinalis Dode(EEEO) in a rat model of acute hepatic necrosis induced by carbon tetrachloride (CCl4).Methods Wistar rats were divided into four groups (control,CCl4,EEEO + CCl4,and Silmyarin + CCl4),the four groups were given intragastrically with normal saline,EEEO for 5 d,respectively.In the last one day,these groups except for control group were injected peritoneally with CCl4.Serum levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP) were detected by automatic biochemistry analyzer.Pathological changes of hepatic tissues were assessed by hematoxylin-eosine (HE) staining.The levels of superoxide dismutase (SOD),catalase (CAT) and malondialdehyde (MDA) in liver homogenate were analyzed using xanthinoxidase and thio-barbituric acid,respectively.Results Compared the ALT [(345.4 ±51.6)U/ml] and AST [(621.7 ± 143.5) U/ml)] of CCl4 group with ALT [(41.1 ± 2.2) U/ml] and AST [(85.2 ± 22.2) U/ml] of control group,the serum levels of ALT and AST in the CCl4 group were increased significantly (P ＜ 0.05).HE staining of liver tissue,the degeneration and necrosis were implicated to the whole hepatic lobules in the CCl4 group.In EEEO + CCl4 group,compared the ALT [(308.1 ± 44.6) U/ml] and AST [(546.4 ± 131.6) U/ml] of low dose EEEO + CCl4 group with the ALT [(210.6 ±34.5) U/ml] and AST [(379.3 ± 112.3) U/ml] of high dose EEEO +CCl4 group the serum levels of ALT and AST were decreased significantly in low dose EEEO + CCl4 group (P ＜0.05).The denaturation and necrosis of hepatic lobules,the level of SOD,CAT were increased and MDA decreased (P ＜ 0.05) inendochylema.Concluslons EEEO can significantly relieve the CCl4-induced hepatonecrosis.The role may be related to anti-lipid peroxidation.

Simultaneous Analysis of Thirteen Bioactive Components in Evodia rutaecarpa and Its Varieties by HPLC-DAD-MS / 中草药·英文版

Objective To control the quality of Evodia rutaecarpa better. Methods An HPLC-DAD-MS/MS method was established for the rapid and efficient identification of bioactive constituents and for simultaneous quantitative analysis of four bioactive ingredients including evodiamine, rutaecarpine, dehydroevodiamine, and evodin in E.rutaecarpa, which was applied to evaluating eight samples of E. rutaecarpa and its varieties from different areas.Results Thirteen potentially bioactive constituents including one flavonoid glycoside, one limonin, four indoloquinazoline alkaloids, and seven quinolone alkaloids were identified in all samples and the contents of dehydroevodiamine, evodine, evodiamine, and rutaecarpine varied widely from 0.10% to 0.51%, 0.49% to 3.12%,0.07% to 1.56%, and 0.10% to 0.69%, respectively. Conclusion This method is found to be convenient, fast,accurate, and it is facilitated to improve the quality control standard of E. rutaecarpa and related products.

X-ray crystallographic analysis and revision of NMR spectral assignments for rhetsinine / 中国药科大学学报

Rhetsinine has been isolated from the fruits of Evodia rutaecarpa ( Juss. ) Benth. ~1H NMR and ~(13)C NMR assignments reported previously for rhetsinine were revised on the basis of UV, IR, ESI-MS,~1H NMR, ~(13)C NMR and X-ray crystallographic analysis.

Optimization Research on Technology of Extracting Evodia rutaecarpa Juice——Crude Drug Processing Adjuvant of Yuhuanglian / 中国中医药信息杂志

Objective To discuss the best extracting technology of Evodia rutaecarpa juice——crude drug processing adjuvant of Yuhuanglian with water.Methods The content of 9 samples extracted according to orthogonal tests were mensurated by UV-spectrophotometry,and the extract methods were estimated with the content of total alkaloid in Evodia rutaecarpa as index.Results The technology of A3B2C1 that was marinated 60 minutes,added the water with 12,9.6 folds,extracted for 40 minutes,and extracted 2 times,which obtained the highest content of total alkaloid.Conclusion This test offered a pressing,feasible and best extracting technology of preparation of crude drug processing adjuvant of Yuhuanglian——Evodia rutaecarpa juice.

Comparison of components in Coptis chinensis processed with various quantity of Evodia rutaecarpa / 中草药

Object To investigate the berberine in Coptis chinensis Franch. processed with various quantity of Evodia rutaecarpa (Juss.) Benth. and the components absorbed from E. rutaecarpa. Methods Taking berberine, evodiamine and rutaecarpine as targets, HPLC method was used to determine the components of C. chinensis, E. rutaecarpa, C. chinensis processed with 10%, 20%, 30%, 40%, 50% E. rutaecarpa. Results The content of berberine in C. chinensis processed with E. rutaecarpa decreased, and that of C. chinensis processed with 20% and 30% E. rutaecarpa was higher than the rest. C. chinensis processed with E. rutaecarpa absorbed the components of E. rutaecarpa really. Conclusion The suitable quantity of E. rutaecarpa is 20% when processing for C. chinensis.

Contens and scavenging effect on hydroxyl radical of flavonoids in Evodia rutaecarpa / 中成药

AIM: To determine the contents of flavonoids and its scavenging effect of flavonoids in Evodia rutaecarpa on hydroxyl radical. METHODS: The flavonoids were determined by Al(NO_3)_3-NaNO_2 spectrophotography method, with sodium salicylate captured the hydroxyl radical based on Fenton reaction to make development, its absorbance was measured at 510 nm and adopted as clearance of scavenging effect on hydroxyl radical. RESULTS: The contents of flavonoids were within 29.39 mg?g -1 - 59.64 mg?g -1 from Evodia rutaecarpa and its different processed product. CONCLUSION: The flavonoids in Evodia rutaecarpa showed better effect on scavenging hydroxyl radical.