Study on sesquiterpenoids from Aquilaria sinensis / 中草药

Objective: To investigate the sesquiterpenoid constituents from Aquilaria sinensis. Methods: The chemical constituents were isolated and purified by various separation techniques such as silica gel, ODS, Sephadex LH-20, and preparative high- performance liquid chromatography, and their structures were determined according to their physicochemical properties, MS, 1D, and 2D NMR. The antibacterial activity of the obtained compounds against methicillin-resistant Staphylococcus aureus was tested by 96-well plate microdilution method. Results: Seven sesquiterpenoids were obtained from 95% ethanol aqueous extract of Aquilaria sinensis and their structures were identified as (+)-4a,5-dimethyl-3-(prop-1-en-2yl)-octahydronaphthalene-2β,8a-diol (1), baimuxinic acid (2), baimuxinol (3), vetaspira-2(11),6-dien-14-al (4), baimuxinal (5), (-)-10-epi-γ-eudesmol (6), and 9β-hydroxyl-α-agarofuran (7). The minimum inhibitory concentration (MIC) of compound 1 against methicillin-resistant Staphylococcus aureus was 210 μmol/L. Conclusion: Compound 1 is a new compound named as 2β,8aα-dihydroxy-11-en-eremophilane, which has a good inhibitory effect against methicillin-resistant Staphylococcus aureus.

Spectrum-effect relationship of dryness of Atractylodis Rhizoma volatile oil based on grey relational grade and orthogonal partial least squares analysis / 中草药

Objective To study the spectrum-effect relationship between GC-MS fingerprint of the volatile oil of Atractylodis Rhizoma and the dryness effect of Atractylodis Rhizoma, and clarify the main dryness components of Atractylodis Rhizoma. Methods The volatile oil of different batches of Atractylodis Rhizoma (S1-S9) were analyzed by GC-MS. Using the daily drinking amount of water, the viscosity of whole blood, and the content of aquaporin 2 (AQP2) in the kidney of rats as the indexes of dryness effect of Atractylodes Rhizome, the spectrum-effect relationship was analyzed by the combination of grey relational grade and orthogonal partial least square method. Results The dryness effect of Atractylodis Rhizoma was the combined action of multiple components, and the peaks of the major contribution to dryness effect were 31, 28, and 33, respectively. The corresponding components of the three peaks were β-eudesmol, (-)-aristolene, and bulnesol, which were identified by the spectral library retrieval analysis. Conclusion The main dryness effect components of Atractylodis Rhizoma are β-eudesmol, (-)-aristolene, and bulnesol, which provide a reference for investigating the material basis of dryness effect of Atractylodis Rhizoma.

Effects of copper stress on accumulation of three medicinal compositions and expression of two key enzyme genes in biosynthesis of Atractylodes lancea / 中草药

Objective: To explore the effect on the accumulation of medicinal compositions β-eudesmol, atractylon, atractylodin and key enzyme genes 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS) expression in biosynthesis of Atractylodes lancea under copper stress. Methods: Under copper stress, the expression of key enzyme genes HMGR and FPPS in A. lancea was determined by real-time fluorescence quantitative PCR; the content of three medicinal components in A. lancea were determined by HPLC; The correlation analysis was performed with SPSS, and DPS software for grey correlation analysis. Results: When the copper stress concentration was within 100 mg/kg, the expression of FPPS and the content of atractylon in the rhizomes of A. lancea increased slightly. However, when the copper concentration continued to increase, the expression levels of HMGR and FPPS and three medicinal components content of A. lancea showed a different degrees of downward trend. The expression levels of HMGR and FPPS were positively correlated with the content of β-eudesmol, atractylon, and atractylodin (P < 0.05) under copper stress. Grey relational analysis showed that the content of β-eudesmol and atractylon in the rhizomes was significantly correlated with the expression of HMGR and FPPS of A. lancea under copper stress. The expression of FPPS gene had the larger contribution on the composition of β-eudesmol and atractylon. However, the correlation between the content of atractylodin and the expression of these two key enzyme genes was relatively small. Conclusion: This study clarified the change regulation of two key enzyme gene expression and the content of three medicinal compositions, and revealed the relationship between β-eudesmol, atractylon and HMGR and FPPS, the key enzymes in terpene biosynthesis of A. lancea under copper stress. It contributed to the further study of the molecular regulation mechanism of the synthesis of medicinal constituents under copper stress and provided a theoretical basis for improving the quality of A. lancea.

First record of the chemical composition of essential oil of Piper bellidifolium, Piper durilignum, Piper acutilimbum and Piper consanguineum from the Brazilian Amazon forest / Primeiro registro da composição química de óleos essenciais de Piper bellidifolium, Piper durilignum, Piper acutilimbum e Piperconsanguineumda Floresta Amazônica no Brasil

Piper bellidifolium, Piper durilignum, Piper acutilimbum and Piper consanguineum are bushes that occur in the Amazon and are morphologically similar. With the aim of analyzing the chemical profile of the volatile constituents of these species, essential oils from the leaves were obtained through steam distillation and analyzed using gas chromatography-flame ionization detection (GC-FID) and gas chromatograph coupled to a mass spectrometer (GC-MS). The chemical analysis enabled the identification of 95 compounds representing 96.3 ± 0.6% of the P. bellidifolium oil, 95.5 ± 0.71% of the P. durilignum oil, 98.0 ± 1.0% of the P. acutilimbum oil and 96.1 ± 2.1% of the P. consanguineum oil. Although sesquiterpenes were the predominant chemical class in the oils of the four species, qualitative and quantitative differences were found in their chemical composition. The major constituents were (E)-nerolidol (20.3 ± 0.4%) in the P. bellidifolium oil, germacrene D (11.1 ± 0.3%) in the P. durilignum oil, and -eudesmol in both the P. consanguineum (18.6 ± 0.5%) and P. acutilimbum (7.5 ± 0.4%) oils. Despite their morphological similarity, a principal component analysis (PCA) of the GC-MS data clearly separated the four species according to the chemical profile of the essential oil extracted from their leaves.

Piper bellidifolium, Piper durilignum, Piper acutilimbum e Piper consanguineum são arbustos que ocorrem na Amazônia e são morfologicamente similares. Com o intuito de analisar o perfil químico dos constituintes voláteis dessas espécies, óleos essenciais das folhas foram obtidos por hidrodestilação e analisados por cromatografia gasosa - detector por ionização de chama (CG-FID) e cromatografia gasosa acoplada a espectrometria de massa (CG-EM). A análise química permitiu identificar 95 compostos, representando 96.3 ± 0.6% do óleo de P. bellidifolium; 95.5 ± 0.71% de P. durilignum; 98.0 ± 1.0% de P. acutilimbum e 96.1 ± 2.1% de P. consanguineum. Apesar dos óleos das quatro espécies terem sesquiterpeno como classe química predominante, diferenças qualitativas e quantitativas em sua composição química foram observadas. Os principais componentes encontrados foram: (E)-nerolidol (20.3 ± 0.4%) em P. bellidifolium; germacreno D (11.1 ± 0.3%) em P. durilignum; e -eudesmol nos óleos de P. consanguineum (18.6 ± 0.5%) e P. acutilimbum (7.5 ± 0.4%). Apesar da similaridade morfológica entre as espécies, uma análise de componentes principais (PCA) dos dados de CG-EM claramente separou as quatro espécies quanto ao perfil químico do óleo essencial extríado de suas folhas.

Composición química del aceite esencial de hojas de hedyosmum translucido cuatrec., chloranthaceae (granizo) / Chemical composition of essential oil leaves hedyosmum translucidum cuatrec., chloranthaceae (granizo)

En el presente trabajo se realizó el análisis de la composición del aceite esencial de hojas de “Granizo”, Hedyosmum translucidum Cuatrec., Chloranthaceae, con certificado de identificación del Herbario PSO, recolectado en el corregimiento de Santa Bárbara, Pasto, Colombia. Este género es reportado con un índice de valor de importancia eco sistémico. El aceite esencial se obtuvo mediante hidrodestilación asistida por microondas, con un rendimiento de 1,2 por ciento. Los componentes presentes en el aceite esencial se identificaron mediante cromatografía de gases acoplada a espectroscopia de masas GC-MS. Los componentes mayoritarios fueron: alfa- eudesmol (11,4 por ciento), germacreno D (8,9 por ciento), trans-beta-cariofileno (7,8 por ciento), elemol (5,8 por ciento) y óxido de cariofileno (5,3 por ciento). Estos compuestos se clasifican como sesquiterpenos. La proporción de los componentes mayoritarios del aceite esencial de H. translucidum presenta diferencias con las reportadas para otras especies del género Hedyosmum en Bolivia, Venezuela y Brasil. Este trabajo constituye un conocimiento inicial para definirse como una especie promisoria.

Essential oil composition of granizoʼs leaves, Hedyosmum translucidum Cuatrec., Chlorantaceae, certified identification Herbarium PSO, collected in town Santa Barbara, Pasto, Colombia were analyzed in this work. This genus is reported with an index value of eco systemic importance. The essential oil was obtained by means of microwave radiation-assisted hydro distillation, extraction yield was 1.2 percent. The components in the essential oil were identified by gas chromatography-mass spectrometry (GC-MS). The most abundant components were: alpha-eudesmol (11.4 percent), germacrene D (8.9 percent), trans-beta-caryophyllene (7.8 percent), elemol (5.8 percent) and caryophyllene oxide (5.3 percent). These compounds are classified as sesquiterpenes. The observed relative amounts of the most abundant constituents of H. translucidum, were different to those reported for other species of genus Hedyosmum in Bolivia, Venezuela and Brazil. This paper is a starting knowledge to be defined as a promising species.

Citotoxicidad/capacidad antiradicalaria in-vitro y caracterización estructural por GC-MS/1H-13C-RMN de los aceites esenciales de hojas de árboles joven/adulto de annona purpurea Moc. & Sessé ex Dunal de Repelón (Atlántico, Colombia) / [in-vitro Cytotoxicity/Anti-Radical Capability and structural characterization by GC-MS/1H13C-NMR of theleaves essential oils of young/old trees of Annona purpurea Moc. & Sessé ex Dunal from Repelón (Atlántico,Colombia]

Annona purpurea es una planta empleada en etnomedicina, en la región norte de Colombia, que no tiene reportes científicos relacionados con la composición de su AE y sus actividades biológicas. En este trabajo, los AE de hojas de árboles joven/adulto se caracterizaron por GC-MS y RMN y se evaluaron sus propiedades antiradicalarias/citotóxicas. Los AE de hojas de árboles joven/adulto estuvieron representados por beta-eudesmol (68.9 por ciento) y alfa-eudesmol (16.8 por ciento), y germacreno D (55.6 por ciento) y biciclogermacreno (20.3 por ciento), respectivamente. El análisis por RMN mostró las señales distintivas de los constituyentes mayoritarios identificados. Los valores de TAA (mmol Trolox®/kg SE) obtenidos por los AE de árboles joven/adulto fueron 165 +/- 8 y 602 +/- 38. Los AE de árboles joven/adulto evaluados sobre linfocitos humanos fueron “moderadamente tóxicos” con valores de CL50 (ug/mL) de 145.5 +/- 0.7 y 346 +/- 8. Finalmente, la citotoxicidad en eritrocitos humanos reveló que el AE de árbol adulto no fue hemolítico (CL50 > 1000 μg/mL, 4.3 +/- 0.6 por ciento); mientras que, el AE de árbol joven fue hemolítico (CL50 490 +/- 48 ug/mL).

Annona purpurea is a plant used in ethnomedicine in the northern region of Colombia, which has no scientific reports on the composition of their essential oil (EO) and biological activities. In this work, the leaves EO of young/old trees were characterized by GC- MS and NMR, and their antiradical/cytotoxic properties were evaluated. beta-Eudesmol (68.9 percent) and alpha-eudesmol (16.8 percent), and germacrene D (55.6 percent) and bicyclogermacrene (20.3 percent), were the representative compounds of the leaves EO of young/old trees, respectively. The NMR analysis showed the distinctive signals of the main constituents identified. The TAA values (mmol Trolox®/kg ES) obtained from the EO of young/old trees were 165 +/- 8 and 602 +/- 38. The EO of young/old trees evaluated on human lymphocytes were “moderately toxic” with LC50 (μg/mL) of 145.5 +/- 0.7 and 346 +/- 8. Finally, the cytotoxicity in human erythrocytes revealed that the old tree EO was not haemolytic (LC50 > 1000 ug/mL, 4.3 +/- 0.6 percent); while the young tree EO was hemolytic (LC50 490 +/- 48 ug/mL).

Determination of β-Eudesmol in Cortex Magnoliae Officinalis by Capillary Gas Chromatography / 中国药师

Objective: To establish a quantitative determination method for β-eudesmol in Cortex Magnoliae Officinalis by GC. Methods:β-Phenethanol was used as the internal standard substance;the column was a Zebron ZB-WAX capillary column ( 60 m × 320 μm,0. 5μm) with the column temperature of 200℃;the detector was FID and the vaporizer temperature was 250℃; the carrier gas was nitrogen with the flow rate of 1. 3 ml · min-1 and the split ratio was 4 ∶1. Results: The linear range of β-eudesmol was 0. 015 1-0. 271 2 mg·ml-1(r=0. 999 8);the average recovery was 99. 28%(RSD =1. 17%, n=6). Conclusion:The method is simple and accurate with good reproducibility, which can be used for the quality control of medicinal materials and decoction pieces of Cortex Magnoliae Officinalis.

Determination of atractylodin, atractylon, and β-eudesmol and study on characteristic spectrum of Atractylodis Rhizoma / 中草药

Objective: To study the content determination of atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma by GC and characteristic spectrum in order to provide a scientific basis for the quality control. Methods: Using GC and Agilent HP-5 capillary column, taking nitrogen as carrier gas, FID as detector, temperature programming, split ratio, injection port temperature: 250℃, detector temperature: 250℃, column temperature: 130℃; The contents of atractylodin, atractylon and β-eudesmol of 25 samples between Chengde and purchased from other markets were determined by external standard method. The characteristic spectrum was set up and the similarity was analyzed by Estimating System of Similarity on the Chinese Medicine Fingerprint Chromatogram. Results: The determination method and characteristic spectrum by GC for atractylodin, atractylon, and β-eudesmol in Atractylodis Rhizoma were established. Nine characteristic peaks were identified; The linear range of β-eudesmol was 20.00 - 406.10 μg/mL (r = 0.999 9), and the average recovery was 100.75%, RSD = 1.17% (n = 6), and the limit of detection was 0.12 ng. The linear range of atractylon was 35.00 - 348.70 μg/mL (r = 0.999 5), and the average recovery was 99.84%, RSD = 1.29% (n = 6), and the limit of detection was 0.04 ng; The linear range of atractylodin was 16.46 - 329.30 μg/mL (r = 0.999 6), and the average recovery was 100.12%, RSD = 0.88% (n = 6), and the limit of detection was 0.06 ng. Conclusion: The concent determination and characteristic spectrum method of atractylodin, atractylon, and β-eudesmol established by this study are sensitive, simple, stability, which could make the determination result accurate and reliable.

GC-MS fingerprint of in vitro anti-gastric cancer active parts from roots of Uygur medicine Ferula ferulaeoides / 中草药

Objective: To preliminarily ascertain the anti-gastric cancer active parts from the roots of Ferula ferulaeoide and to study its GC-MS fingerprint Methods: The inhibitory effects of different extraction from the roots of F. ferulaeoide on the cell proliferation of SGC-7901 cells were determined with MTT colorimetric method. GC-MS fingerprint of in vitro anti-gastric cancer active parts from F. ferulaeoide was investigated and analyzed by GC-MS and principal component analysis (PCA). Results: The chloroform extract showed the best inhibition on the growth of SGC-7901 cells. The method on GC-MS fingerprint of in vitro anti-gastric cancer active parts from F. ferulaeoide was established, showing 28 common characteristic peaks. The PCA demonstrated that the common peaks 1, 2, 4, 6, 7, 8, 9, 12, 21, 22, and 23 were connected closely with the in vitro anti-gastric cancer activity, and the seven compounds were 3-methoxy-1,2-propanediol, D-limonene, L-borneol acetate, terpinyl acetate, 1,5,9-undecatriene, 2,6,10-trimethyl, α-cedrene, and a-bergsmotene, β-cedrene, 8-epi-γ-eudesmol, γ-eudesmol, hinesol. Totally 28 common compounds of 10 batches of samples accounted for over 92% of the volatile contents, and the similarity of the GC-MS fingerprints from the 10 batches of samples was over 0.90. Conclusion: The chloroform extract from the roots of F. ferulaeoides with potential inhibitory effect on gastric cancer SGC-7901 cells is tentatively confirmed, and needs further to verify by animal cells in vivo. The method of GC-MS fingerprinting is rapid, simple, and accurate with good reproducibility and stability, and can be used to control the quality of active parts of F. ferulaeoides.

Traditional Herbal Medicine for the Control of Tropical Diseases

Throughout history, traditional herbal medicine has afforded a rich repository of remedies with diverse chemical structures and bioactivities against several health disorders. A common issue of herbal medicine is the limitation of information on their pharmacological activities and their active constituents. Traditionally, the use of herbal medicine has been based on empirical treatment and passed on from generation to generation with information available only in local journals. This prevents several herbal medicines from being developed to their full potential. The presentation will focus on research and development of Atractylodes lancea (Thunb) DC. (AL: family Compositae) as a potential chemotherapeutic for cholangiocarcinoma (CCA), the bile duct cancer commonly found in Southeast Asia. The dried rhizome of AL is a medicinal plant used in Chinese (“Cang Zhu”), Japan (“So-jutsu”) and Thai (“Khod-Kha-Mao”) traditional medicine for its various pharmacological properties including anticancer, anti-inflammation and antimicrobial activities, activities on central nervous, cardiovascular, and gastrointestinal systems. The major constituents in the essential oils from AL rhizome are β-eudesmol, hinesol and atractylon. Preliminary investigation has demonstrated its promising anti-CCA activity both <i>in vitro</i> and animal (Opisthorchis viverrini/dimethylnitrosamine-induced CCA in hamsters and CCA—xenografted nude mice) models with high selectivity index comparing with the standard drug, 5-fluorouracil. It also showed virtually no toxicity with only minimal CNS effects on locomotor activity at the maximum dose of 5,000 mg/kg body weight. Studies are underway to identify active constituent(s) which contribute to anti-CCA activity as well as its pharmacokinetic and pharmacodynamic properties. The main research interest of my research group is the discovery and development of traditional herbal medicine for the treatment of two important tropical diseases, cholangiocarcinoma and malaria. As the time is quite limited, I am going to give you the summary of the conceptual framework and highlight some important findings which will illustrate how different approaches have been used or applied for the discovery of the promising candidates for these two diseases.

Traditional Herbal Medicine for the Control of Tropical Diseases

Throughout history, traditional herbal medicine has afforded a rich repository of remedies with diverse chemical structures and bioactivities against several health disorders. A common issue of herbal medicine is the limitation of information on their pharmacological activities and their active constituents. Traditionally, the use of herbal medicine has been based on empirical treatment and passed on from generation to generation with information available only in local journals. This prevents several herbal medicines from being developed to their full potential. The presentation will focus on research and development of Atractylodes lancea (Thunb) DC. (AL: family Compositae) as a potential chemotherapeutic for cholangiocarcinoma (CCA), the bile duct cancer commonly found in Southeast Asia. The dried rhizome of AL is a medicinal plant used in Chinese (“Cang Zhu”), Japan (“So-jutsu”) and Thai (“Khod-Kha-Mao”) traditional medicine for its various pharmacological properties including anticancer, anti-inflammation and antimicrobial activities, activities on central nervous, cardiovascular, and gastrointestinal systems. The major constituents in the essential oils from AL rhizome are β-eudesmol, hinesol and atractylon. Preliminary investigation has demonstrated its promising anti-CCA activity both in vitro and animal (Opisthorchis viverrini/dimethylnitrosamine-induced CCA in hamsters and CCA—xenografted nude mice) models with high selectivity index comparing with the standard drug, 5-fluorouracil. It also showed virtually no toxicity with only minimal CNS effects on locomotor activity at the maximum dose of 5, 000 mg/kg body weight. Studies are underway to identify active constituent(s) which contribute to anti-CCA activity as well as its pharmacokinetic and pharmacodynamic properties. The main research interest of my research group is the discovery and development of traditional herbal medicine for the treatment of two important tropical diseases, cholangiocarcinoma and malaria.As the time is quite limited, I am going to give you the summary of the conceptual framework and highlight some important findings which will illustrate how different approaches have been used or applied for the discovery of the promising candidates for these two diseases.

Promotion of HMGR activation and β-eudesmol biosynthesis in Atractylodes lancea suspension cell culture by hydrogen peroxide-mediated endopytic fungal elicitor / 中草药

Objective: To investigate the effect of endophytic fungal elicitor on key enzyme activity, inducing pathway and mechanism involved in the secondary metabolites of Atractylodes lancea. Methods: NADPH oxidase, HMGR activities, the concentration of hydrogen peroxide (H2O2) and β-eudesmol were determined by the co-culture of endophytic fungal elicitor and A. lancea suspension cell. Results: NADPH oxidase activity was notably enhanced by Fusarium sp5 elicitor which could induce oxidative burst, significantly promote H2O2 accumulation, and activate HMGR in the sesquiterpenoids metabolic pathway. Compared with the control, the yield of β-eudesmol increased 257.6% and reached 66.59 μg/g. CAT and DPI could inhibit the HMGR activity and β-eudesmol biosynthesis in A. lancea cell induced by Fusarium sp5 elicitor. Exogenous H2O2 also induced HMGR and promoted the β-eudesmol biosynthesis. Conclusion: H2O2 is necessary to induce β-eudesmol synthetic signal molecule by activating the HMGR.

Chemical Constituents of the Essential Oil of Laggera pterodonta (DC.) Sch. Bip. From North-Central Nigeria.

The aerial part of Laggera pterodonta plant found in North-Central part of Nigeria was hydrodistilled and the volatile oil subjected to GCMS analysis. 23 components were identified in the essential oil of which n-Triacontane was the major constituent (~43%). Other major volatile constituents include, Dimethoxydurene (~9%), Caryophyllene oxide (~7%), Linoleoyl chloride (~7%), oleic acid (~4%), gamma-Eudesmol (~4%), 2,6,10-trimethylundeca-1,3,diene (~3%) and n-Dotriacontane (~3%). This is the first time that n-triacontane is being found as the major constituent of an essential oil, and also the first time the composition of the essential oil of Laggera pterodonta from North-central Nigeria is being reported.

Determination of ?-eudesmol in atractylis oil from Rhizomes of Atractylodes Lancea (THUNB.)DC.by gas chromatography / 中成药

AIM: To establish the determination of ? eudesmol in atractylis oil from Rhizomes of Atractylodes Lancea (THUNB.) DC. by gas chromatography. METHODS: Pentadecanol had been used as the internal standard substance in internal standard method. The GC system consisted of capillary column, 10%SF 30 as the stationary phase, nitrogen as the carrier gas, and FID as the detector. RESULTS: Both ? eudesmol in essential oil and pentadecanol had got satisfactory separation under the chromatographic condition. The mean recovery of ? eudesmol was 99.60%, and RSD was 1.30%. CONCLUSIONS: The method is sensitive, accurate and reproducible, and it can be used to control the quality of the essential oil from Rhizomes of Atractylodes Lancea (THUNB.) DC.

Quality study of multi-chemical compositions in Ermiao Pills / 中成药

AIM: To establish methods of determing ?-eudesmol in atractylis oil from Ermiao Pills(Rhizoma Atractylodis and Cortex Phellodendri Chinensis) by gas chromatography and determing and comparing the chemical compositions between atractylis oil and poultice of Ermiao Pills by decocting separately and together. METHODS: ?-eudesmol content in Ermiao Pills decocting separtely and together was determined and compared by GC using paeonol as the internal standard substance,and contents of berberine,palmatine,jatrorrhizine in Ermiao Pills decocting separately and together were determined and compared by HPLC. RESULTS: The ?-eudesmol content in the volatile oil of decoction together was more than that of the decoction separately.But there was no remarkable difference.Berberine,palmatine,jatrorrhizine contents of decoction separately were more than that of the together. CONCLUSION: The method of determing ?-eudesmol in Ermiao Pills is established.And it has mutual effect in compatibility with Cortex Phellodendri Chinensis and Rhizoma Atractylodis.