Sephadex LH-20 Column Research Articles

A new cembranoid diterpene from Olibanum

Five compounds were isolated from the alcohol extract of Olibanum by MCI, silica gel, ODS, and Sephadex LH-20 column chromatographies and preparative high-performance liquid chromatography(HPLC). On the basis of spectral data and literature data, the compounds were identified as:(1S,3R,4S,7R,11S,12R)-1:12,4:7-diepoxisonane-8(19)-ene-3,11-diol(1), boscartin A(2),(+)-resinolin(3),(+)-5-hydroxy-3,4-dimethyl-5-pentylfuran-2(5H)-one(4), and acerogenin A(5). Compound 1 is a new compound, and compounds 3-5 were isolated from Olibanum for the first time. The structure of compound 1 was determined by spectroscopic analysis and single-crystal X-ray diffraction. Compounds 1 and 2 were tested for PC12 neurotoxicity, and the results showed that they were both safe compounds.

Evaluation of Scopoletin from Penicillium janthinellum Snef1650 for the Control of Heterodera glycines in Soybean.

Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) is responsible for causing a major soybean disease globally. The fungal strain Penicillium janthinellum Snef1650 was evaluated against H. glycines. However, the effective determinants of the P. janthinellum strain are unknown. By performing pot experiments, a functioning compound was isolated from P. janthinellum Snef1650 through organic solvent extraction, semi-preparative HPLC, Sephadex LH-20 column chromatography, and silica gel column chromatography, and the isolated compound was identified to be scopoletin through 1H NMR, 13C NMR, and HPLC–MS. The pot experiments indicated that the treatment of soybean seeds with scopoletin drastically reduced the SCN population. The field experiments performed in 2017 and 2018 revealed that scopoletin decreased over 43.7% juveniles in the roots and over 61.55% cysts in the soil. Scopoletin treatment also promoted soybean growth and improved its yield, with an increase in plot yield by >5.33%. Scopoletin obtained from P. janthinellum Snef1650 could be used as an anti-H. glycines biocontrol agent.

Chemoenzymatic and Protecting-Group-Free Synthesis of 1,4-Substituted 1,2,3-Triazole-α-d-glucosides with Potent Inhibitory Activity toward Lysosomal α-Glucosidase.

α-Glucosyl triazoles have rarely been tested as α-glucosidase inhibitors, partly due to inefficient synthesis of their precursor α-d-glucosylazide (αGA1). Glycosynthase enzymes, made by nucleophile mutations of retaining β-glucosidases, produce αGA1 in chemical rescue experiments. Thermoanaerobacterium xylanolyticus glucosyl hydrolase 116 β-glucosidase (TxGH116) E441G nucleophile mutant catalyzed synthesis of αGA1 from sodium azide and pNP-β-d-glucoside (pNPGlc) or cellobiose in aqueous medium at 45 °C. The pNPGlc and azide reaction product was purified by Sephadex LH-20 column chromatography to yield 280 mg of pure αGA1 (68% yield). αGA1 was successfully conjugated with alkynes attached to different functional groups, including aryl, ether, amine, amide, ester, alcohol, and flavone via copper-catalyzed azide-alkyne cycloaddition (CuAAC) click chemistry reactions. These reactions afforded the 1,4-substituted 1,2,3-triazole-α-d-glucoside derivatives AGT2-14 without protection and deprotection. Several of these glucosyl triazoles exhibited strong inhibition of human lysosomal α-glucosidase, with IC50 values for AGT4 and AGT14 more than 60-fold lower than that of the commercial α-glucosidase inhibitor acarbose.

Inflammatory and Cytotoxic Activities of Abietane Terpenoids from Nepeta bracteata Benth.

Nepeta bracteata Benth. is used clinically to treat tracheal inflammation, coughs, asthma, colds, fevers, adverse urination, and other symptoms, along with functions in clearing heat and removing dampness. However, there have been few studies characterizing the material basis of its efficacy. Therefore, the aim of this study was to screen for compounds with anti-inflammatory activities in N. bracteata Benth. Using silica gel, ODS C18, and Sephadex LH-20 column chromatography, as well as semipreparative HPLC, 10 compounds were separated from N. bracteata Benth. extract, including four new diterpenoids (1–4), one amide alkaloid (5), and five known diterpenoids (6–10). The structures of all the isolates were elucidated by HR-ESI-MS, NMR, and CD analyses. Using lipopolysaccharide (LPS)-stimulated RAW 264.7 cells, we investigated the anti-inflammatory activities of compounds 1–10. It is worth noting that all were able to inhibit nitric oxide (NO) production with IC50 values < 50 μM and little effect on RAW 264.7 macrophage viability. Compounds 2 and 4 displayed remarkable inhibition with IC50 values of 19.2 and 18.8 μM, respectively. Meanwhile, screening on HCT-8 cells demonstrated that compounds 2 and 4 also had moderate cytotoxic activities with IC50 values of 36.3 and 41.4 μM, respectively, which is related to their anti-inflammatory effects.

Prenylated Flavonoids and C-15 Isoprenoid Analogues with Antibacterial Properties from the Whole Plant of Imperata cylindrica (L.) Raeusch (Gramineae).

The local botanical Imperata cylindrica in Cameroon was investigated for its antibacterial potency. The methanol extract afforded a total of seven compounds, including five hitherto unreported compounds comprising three flavonoids (1–3) and two C-15 isoprenoid analogues (4 and 5) together with known derivatives (6 and 7). The novelty of the flavonoids was related to the presence of both methyl and prenyl groups. The potential origin of the methyl in the flavonoids is discussed, as well as the chemophenetic significance of our findings. Isolation was performed over repeated silica gel and Sephadex LH-20 column chromatography and the structures were elucidated by (NMR and MS). The crude methanol extract and isolated compounds showed considerable antibacterial potency against a panel of multi-drug resistant (MDR) bacterial strains. The best MIC values were obtained with compound (2) against S. aureus ATCC 25923 (32 µg/mL) and MRSA1 (16 µg/mL).

Bibenzyls from Dendrobium officinale

Fifteen bibenzyls were isolated and purified from the ethyl acetate extract of the stems of Dendrobium officinale by macroporous resin, MCI, silica gel, Sephadex LH-20, and ODS column chromatographies, as well as preparative thin-layer chromatography and preparative HPLC. The structures of compounds were identified according to the spectra data of ~1H-NMR, ~(13)C-NMR, and MS, and the physical and physiochemical properties: dendrocandin X(1), 3,4'-dihydroxy-4,5-dimethoxybibenzyl(2), 6″-de-O-methyldendrofindlaphenol A(3), 3,4-dihydroxy-4',5-dimethoxybibenzyl(4), dendrosinen B(5), 3,4,4'-trihydroxy-5-methoxybibenzyl(6), 3,3'-dihydroxy-4,5-dimethoxybibenzyl(7), 3,4'-dihydroxy-5-methoxybibenzyl(8), moscatilin(9), gigantol(10), 4,4'-dihydroxy-3,5-dimethoxybibenzyl(11), 3,4',5-trihydroxy-3'-methoxybibenzyl(12), 3-O-methylgigantol(13), dendrocandin U(14), and dendrocandin N(15). Compound 1 was a novel compound. Compound 2 was isolated from Dendrobium species for the first time. Compounds 3-7 were isolated from D. officinale for the first time.

Chemical constituents of sesquiterpenes from Chloranthus multistachys

With repeated silica gel, octadecyl silica(ODS), and Sephadex LH-20 column chromatography, normal-phase and reverse-phase high performance liquid chromatography(HPLC), etc., a pair of new enantiomers and 5 known compounds were separated from the 95% ethanol extract of Chloranthus multistachys. These compounds were identified by the nuclear magnetic resonance spectroscopy(including 1 D-NMR and 2 D-NMR), single-crystal X-ray diffraction, circular dichroism(CD) spectroscopy, mass spectrometry(MS), and some other methods as(1R,4R,5R,8S,10R)-chloraeudolide H(1 a),(1S,4S,5S,8R,10S)-chloraeudolide H(1 b), hydroxyisogermafurenolide(2), 4α-hydroxy-5α,8β(H)-eudesm-7(11)-en-8,12-olide(3), chloraniolide A(4), chlorantene D(5), 4α,8β-dihydroxy-5α(H)-eudesm-7(11)-en-8,12-olide(6). Compounds 1 a and 1 b are a pair of new eudesmane-type sesquiterpene enantiomers, and compounds 2-4 were isolated from C. multistachys for the first time.

Anti-Neuroinflammatory and Anti-Inflammatory Activities of Phenylheptatriyne Isolated from the Flowers of Coreopsis lanceolata L. via NF-κB Inhibition and HO-1 Expression in BV2 and RAW264.7 Cells.

Aging is associated with immune disregulation and oxidative stress which lead to inflammation and neurodegenerative diseases. We have tried to identify the anti-neuroinflammatory and anti-inflammatory components of Coreopsis lanceolata L. The dried flowers of C. lanceolata were extracted with 70% EtOH, and the obtained extract was divided into CH2Cl2, EtOAc, n-BuOH, and H2O fractions. The CH2Cl2 fraction was separated using silica gel and C-18 column chromatography to yield phenylheptatriyne (1), 2′-hydroxy-3,4,4′-trimethoxychalcone (2), and 4′,7-dimethoxyflavanone (3). Additionally, the EtOAc fraction was subjected to silica gel, C-18, and Sephadex LH-20 column chromatography to yield 8-methoxybutin (4) and leptosidin (5). All the compounds isolated from C. lanceolata inhibited the production of nitric oxide (NO) in LPS-induced BV2 and RAW264.7 cells. In addition, phenylheptatriyne and 4′,7-dimethoxyflavanone reduced the secretion of inflammatory cytokines, tumor necrosis factor alpha (TNF-α), and interleukin (IL)-6. Among them, phenylheptatriyne was significantly downregulated in the expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2). Subsequently, phenylheptatriyne also effectively inhibited nuclear factor-kappa B (NF-κB) activation in LPS-stimulated BV2 and RAW264.7 cells. Based on these results, the anti-neuroinflammatory effect of phenylheptatriyne isolated from C. lanceolata was confirmed, which may exert a therapeutic effect in treatment of neuroinflammation-related diseases.

Erythrina alkaloids from stems of Erythrina corallodendron

This study aims to investigate Erythrina alkaloids from the stems of Erythrina corallodendron. Eighteen Erythrina alkaloids were isolated from the 95% ethanol extract of the stems of E. corallodendron by silica gel,octadecyl silica( ODS),Sephadex LH-20 column chromatography and preparative HPLC. With nuclear magnetic resonance( NMR) spectroscopy and mass spectrometry( MS),their structures were identified as crstanine A( 1),erytharbine( 2),cristamine C( 3),( +)-erystramindine( 4),10,11-dioxoerythraline( 5),8-oxoerythraline( 6),8-oxo-11β-methoxyerythradine( 7),11-methoxyerythradine( 8),( ±)-11-epi-methoxyerythraline( 9),( +)-erythraline( 10),crystamidine( 11),8-oxoerythrinine( 12),( +)-11α-hydroxyerysotrine( 13),erythrinine( 14),erysodine( 15),erysotrine-N-oxide( 16),( +)-erythratidine( 17),erythratine( 18). Compounds 1-4,7,9,11,13,16 and 17 were isolated from E. corallodendron for the first time. Furthermore,the cytotoxic activities of these Erythrina alkaloids were screened by MTT assay. The results showed that all compounds had no obvious cytotoxic activity. The analgesic activities of compounds1,6 and 8 were evaluated using an acetic acid-induced writhing test in mice. The writhing inhibition rates of compounds 1,6 and 8 at20 mg·kg~(-1)( ip) were 69%,70% and 62%,respectively( P&lt;0. 01),indicating they have significant analgesic activity.

Chemical constituents from the leaves of Cinnamomum camphora var. linaloolifera and their anti-inflammatory activities

Thirteen compounds were isolated and purified from the leaves of Cinnamomum camphora by the macroporous resin,silica gel,and Sephadex LH-20 column chromatographies. Those compounds were further identified by IR,UV,MS,and NMR techniques:( 2 S)-1-( 3″,4″-methylenedioxy phenyl)-3-( 2&apos;,6&apos;-dimethoxy-4&apos;-hydroxyphenyl)-propan-2-ol( 1),( 2 R,3 R)-5,7-dimethoxy-3&apos;,4&apos;-methylenedioxy flavanol( 2),9-hydroxysesamin( 3),sesamin( 4),piperitol( 5),kobusin( 6),(-)-aptosimon( 7),acuminatolide( 8),1β,11-dihydroxy-5-eudesmene( 9),lasiodiplodin( 10),vanillin( 11),p-hydroxybenzaldehyde( 12),and p-hydroxybenzoic acid ethyl ester( 13). Compound 1 was a novel compound,and compounds 2,6,7,9 and 10 were isolated from Cinnamomum plants for the first time. Compounds 4,7 and 10 were found to possess good inhibitory effect on IL-6 production in LPS-induced BV2 cells at a concentration of 20 μmol·L-1 in the in vitro bioassay,with inhibition rates of 51. 26% ± 4. 13%,67. 82% ± 3. 77% and85. 81%±1. 19%,respectively.

Examining the Performance of Two Extraction Solvent Systems on Phenolic Constituents from U.S. Southeastern Blackberries

Two common extraction solvent systems, namely acidified aqueous methanol and acidified aqueous acetone, were used to extract blackberry phenolics, and the antioxidant properties of the recovered extracts were compared. The crude extracts were fractionated into low- and high-molecular-weight phenolics by Sephadex LH-20 column chromatography. The hydrophilic-oxygen radical absorbance capacity (H-ORACFL), ferric reducing antioxidant power (FRAP), and the cellular antioxidant activity (CAA) assays were employed as indices to assess antioxidant capacity of the extracts and their respective fractions. The methanolic solvent system displayed a greater efficiency at extracting anthocyanin and flavonol constituents from the blackberries, while the acetonic solvent system was better at extracting flavan-3-ols and tannins. Anthocyanins were the dominant phenolic class found in the blackberries with 138.7 ± 9.8 mg C3G eq./100 g f.w. when using methanol as the extractant and 114.6 ± 3.4 mg C3G eq./100 g f.w. when using acetone. In terms of overall antioxidant capacity of blackberry phenolics, the acetonic solvent system was superior. Though present only as a small percentage of the total phenolics in each crude extract, the flavan-3-ols (42.37 ± 2.44 and 51.44 ± 3.15 mg/100 g f.w. in MLF and ALF, respectively) and ellagitannins (5.15 ± 0.78 and 9.31 ± 0.63 mg/100 g f.w. in MHF and AHF, respectively) appear to account for the differences in the observed antioxidant activity between the two solvent systems.

Structural Characterization and Assessment of Anti-Inflammatory and Anti-Tyrosinase Activities of Polyphenols from Melastoma normale.

Polyphenols, widely distributed in the genus Melastoma plants, possess extensive cellular protective effects such as anti-inflammatory, anti-tyrosinase, and anti-obesity, which makes it a potential anti-inflammatory drug or enzyme inhibitor. Therefore, the aim of this study is to screen for the anti-inflammatory and enzyme inhibitory activities of compounds from title plant. Using silica gel, MCI, ODS C18, and Sephadex LH-20 column chromatography, as well as semipreparative HPLC, the extract of Melastoma normale roots was separated. Four new ellagitannins, Whiskey tannin C (1), 1-O-(4-methoxygalloyl)-6-O-galloyl-2,3-O-(S)-hexahydroxydiphenoyl-β-d-glucose (2), 1-O-galloyl-6-O-(3-methoxygalloyl)-2,3-O-(S)-hexahydroxydiphenoyl-β-d-glucose (3), and 1-O-galloyl-6-O-vanilloyl-2,3-O-(S)-hexahydroxydiphenoyl-β-d-glucose (4), along with eight known polyphenols were firstly obtained from this plant. The structures of all isolates were elucidated by HRMS, NMR, and CD analyses. Using lipopolysaccharide (LPS)-stimulated RAW2 64.7 cells, we investigated the anti-inflammatory activities of compounds 1–4, unfortunately, none of them exhibit inhibit nitric oxide (NO) production, their IC50 values are all > 50 μM. Anti-tyrosinase activity assays was done by tyrosinase inhibition activity screening model. Compound 1 showed weak tyrosinase inhibitory activity with IC50 values of 426.02 ± 11.31 μM. Compounds 2–4 displayed moderate tyrosinase inhibitory activities with IC50 values in the range of 124.74 ± 3.12–241.41 ± 6.23 μM. The structure–activity relationships indicate that hydroxylation at C-3′, C-4′, and C-3 in the flavones were key to their anti-tyrosinase activities. The successful isolation and structure identification of ellagitannin provide materials for the screening of anti-inflammatory drugs and enzyme inhibitors, and also contribute to the development and utilization of M. normale.

Purified antioxidant from the medicinal mushroom Phellinus pini protects rat H9c2 cell against H2 O2 -induced oxidative stress.

In this study, through the combination of AB-8 macroporous resin, Sephadex LH-20 column chromatography and semi-preparative HPLC, an antioxidant component was purified from the crude extract of Phellinus pini, thereby evaluating the cardioprotective effect of the fraction. As a result, total phenolic content of the 60% ethanol elution was increased by 4.8-fold after one run treatment on Sephadex LH-20 chromatography with gradient elution. After semi-preparative HPLC separation, the first peak (PP-S4-1) showed that inhibition ratio of erythrocyte hemolysis was 91.9%, and inhibition ratio of lipid peroxidation was also increased by 87.6%, at 50μg/ml (p<.01). Based on the results of ESI-MS, 1 HNMR, 13 CNMR, and RP-HPLC compared to many published results, PP-S4-1was identified as catechin (MW 290.015, C15 H14 O6 ). The results showed that PP-S4-1 pretreatment made cell viability increased, and the generation of reactive oxygen species (ROS) inhibited. Meanwhile, PP-S4-1 remarkably decreased the fluorescence intensity of Ca2+ , and increased mitochondrial membrane potential (MMP; ΔΨm). In addition, PP-S4-1 could significantly inhibit the decrease of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) activity as well as the increase of MDA content in H9c2 cells induced by H2 O2 . Moreover, pretreatment with PP-S4-1 significantly improved the morphological changes and prevented H2 O2 -induced DNA damage. Therefore, this study clarifies the ability of PP-S4-1 to treat H9c2 cell oxidative stress damage induced by H2 O2 through its antioxidant effect. PRACTICAL APPLICATIONS: This research is not only helpful to elaborate the cardioprotective effect of Phellinus pini but also can contribute to the development of health foods or drug supplements for heart disease in the future. This is the first report dealing with phenolic component and cardioprotective activity of a medicinal mushroom P. pini belonging to the genus Phellinus.

Two new sphingolipids from the stem bark of Synsepalum msolo (Sapotaceae)

Synsepalum msolo commonly known as Bang Bali in Bali-Nguemba, Cameroon is used in traditional medicine against various diseases. The leaves and stem bark extracts were subjected to silica gel and Sephadex LH20 column chromatography to yield pure compounds. The structures of the compounds were determined by detail analysis of NMR and Mass spectroscopic data and by comparison with data reported in the literature. Amongst the isolates, were two new sphingolipids: synsepaloside B (1), synsepaloside C (2), and five known compounds: (+)-catechin (3), (−)-epicatechin (4), myricitrin (5), triacontanol (6), and aurantiamide acetate (7). Compounds 1–5 were screened for their antibacterial and anti-yeast activities on several microorganisms. All the tested compounds exhibited weak antibacterial (MIC ≥ 200 μg/mL) and anti-yeast (MIC > 200 μg/mL) activities as compared to standard: ciprofloxacin 0.468 < MIC >0.234 μg/mL and fluconazole MIC = 0.05 μg/mL, respectively.

Antioxidant activity of Malpighia glabra L., leaves extract

Malpighia glabra. L (Acerola) fruits have wide global medicinal uses due to rich antioxidant contents. Little studies were carried on other parts of plant. Malpighia glabra. L leaves (1.5 kg) was subjected to soxhlet exhaustive extraction with methanol and concentrated under vacuum. The dried extract was defatted with petroleum-ether then fractionated with ethyl acetate and n-butanol 3 successive times respectively. The ethyl acetate fraction (9 gm) was subjected to series of silica gel and Sephadex LH-20 column chromatography. Phytochemical investigation of the Ethyl acetate fraction of Malpighia glabra. L leaves in this study resulted in separation of four pure compounds (Caffeic acid, Chlorogenic acid, Quercetin & Kaempferol). The structures of the separated compounds were assigned based on NMR spectral data and mass spectrometer analysis. The methanolic extract showed high values of total phenolics (96.36 mg GAE/g), flavonoids (30.71 mg QE/g) and higher antioxidant activity towards DPPH radical with IC50 = 49.8(mg/ml).

Triterpenoids from leaves of Ilex guayusa

The chemical constituents from the leaves of Ilex guayusa were investigated. Sixteen triterpenoids were isolated from the 95% ethanol extract of dried leaves of I. guayusa by silica gel, Sephadex LH-20, and ODS column chromatographies and semi-prepa-rative HPLC. Those triterpenoids were identified by NMR, HR-MS, and literature analysis: 3β-hydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13β-olide(1), 3β-hydroxy-24-nor-4(23),12-oleanadien-28-methyl ester(2), oleanolic acid(3), 3β,28-dihydroxy-12-oleanene(4), 2α,3β-dihydroxy-11α,12α-epoxy-24-'nor-olean-4(23)-ene-28,13β-olide(5), ursolic acid(6), 3β,23-dihydroxy ursolic acid(7), 3β,28-dihydroxy-12-ursene(8), 3β-28-nor-urs-12-ene-3,17-diol(9), 3β-hydroxyurs-11-ene-28,13β-olide(10), 13β,28-epoxy-3β-hydroxy-11-ursene(11), 3β-hydroxy-28,28-dimethoxy-12-ursene(12), 3β-hydroxy-24-nor-urs-4(23),12-dien-28-oic acid(13), 3β-hydroxy-24-nor-urs-4(23),12-dien-28-methyl ester(14), 2α,3β-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23)-ene-28,13β-olide(15) and 2α,3β-dihydroxy-11α,12α-epoxy-24-nor-urs-4(23),20(30)-dien-28,13β-olide(16). Compounds 1-2 were new compounds, and compounds 4-5, 7 and 9-16 were isolated from I. guayusa for the first time.

A novel flavonoid with antioxidant activity from Patrinia villosa (Thunb.) Juss

Patrinia villosa (Thunb.) Juss (P. villosa), a perennial herb, is widely used as a medicinal plant in Chinese folk. This study aims to isolate and identify the chemical constituents from P. villosa and evaluate their antioxidant activity. Normal silica column chromatography, ODS silica column chromatography, Sephadex LH-20 column chromatography and semi-preparative HPLC methods were used to obtain a new compound named 3-n-pentadecyl-4’-methoxyluteolin (1) and two known compounds including luteolin-7-O-β-D-glucuronide methyl ester (2) and apigenin-7-O-β-D-glucuronide methyl ester (3). The antioxidant activity of these compounds was determined by DPPH and ABTS methods and the IC50 values were calculated. The IC50 values of ABTS scavenging activity of 1, 2 and 3 were 12.99 ± 0.09 μM, 7.13 ± 0.07 μM and 5.15 ± 0.08 μM, respectively, and the IC50 values of DPPH scavenging activity of 1, 2 and 3 were 51.86 ± 0.41 μM, 23.95 ± 0.71 μM and 25.06 ± 0.65 μM, respectively. All the compounds exhibited good antioxidant activities in vitro.

Diagnostic ion filtering targeted screening and isolation of anti-inflammatory iridoid glycosides from Hedyotis diffusa.

Efficient and targeted screening and isolation of bioactive compounds from complex natural products is still a challenging work. Herein, diagnostic ion filtering based high-performance liquid chromatography-quadrupole time-of-flight-tandem mass spectrometry was firstly developed to screen six main iridoid glycosides from Hedyotis diffusa. Then, online extraction-high-speed counter current chromatography was proposed for targeted enrichment and preparative isolation using ethyl acetate/n-butanol/water (4.5:0.5:5, v/v/v) as solvent system. After that, Sephadex LH-20 column chromatography using methanol as solvent system was selected for further purification of six iridoid glycosides with purities over 98%. They were finally identified as monotropein, desacetylasperuloside acid, asperuloside, 6-O-(Z)-p-coumaroyl scandoside methyl ester, 6-O-(Z)-feruloyl scandoside methyl ester, and 6-O-(E)-p-coumaroyl scandoside methyl ester. And their anti-inflammatory activities were evaluated and confirmed by lipopolysaccharide activated RAW 264.7 macrophages. Obviously, the results provide a scientific basis for the potential applications of H. diffusa, and the developed methodology is efficient and reliable for targeted screening and isolation of bioactive compounds from natural products.

The phytochemical investigation from n-hexane extract of the lichen Roccella montagnei

Introduction: R occella montagnei is widely distributed in subtropical regions. As the continuous study on the hexane extract of Roccella montagnei lichen, the isolation and structural determination of five compounds were addressed.&#x0D; Method: The crude extract was obtained from the dried lichen powder's extraction at room temperature. The n-hexane, n-hexane-ethyl acetate, and ethyl acetate extracts were obtained by the liquid-liquid partition method. The organic compounds were isolated from n-hexane extract by silica gel and Sephadex LH-20 column chromatography. Their chemical structures were identified by the NMR and HR-ESI-MS data analysis and the comparison of their NMR data with the published data.&#x0D; Results: Five compounds were isolated and chemically structural identified, consisting of 3b -hydroxy-7a-methoxystigmast-5-ene (1), sekikaic acid (2), lichenxanthone (3), (+)-6,8-dihydroxy-3-propyl-3,4-dihydroisocoumarin (4), and ar-turmerone (5).&#x0D; Conclusion: To the best of our knowledge, except 3 which was reported from this species for the first time, four isolated compounds left did not known to be present in Roccella genus before.

Gallotannins from Nut Shell Extractives of Camellia oleifera

Camellia nut shell was collected, dried at room temperature and ground to get fine powder. The powder was extracted three times with 95% EtOH, combined, evaporated, and then freeze dried. The crude powder was dissolved in H2O and then sequentially fractionated with n-hexane, CH2Cl2, EtOAc and n-BuOH. A part of EtOAc fraction was chromatographed on a silica gel and on a Sephadex LH-20 columns using MeOH, aqueous MeOH, EtOAc-n-hexane and EtOH-n-hexane to isolate gallotannins. Three gallotannins, 1,2-di-O-galloyl-β-D-glucopyranoside (2), 1,2,6-tri-O- galloyl-β-D-glucopyranoside (3) and 1,2,3,6-tetra-O-galloyl-β-D-glucopyranoside (4), including gallic acid (1), were isolated and elucidated by NMR and Mass spectroscopies. Although nothing new, these gallotannins were first reported from the nut shell extractives of camellia tree (Camellia oleifera C. Abel). This study was to investigate the chemical constituents, especially hydrolysable tannins, of nut shell extractives of Camellia oleifera and to provide basic information for the future chemical utilization of this species.