Sephadex LH-20 Column Research Articles

Study on non-flavonoids chemical constituents from Spatholobi Caulis

To study the non-flavonoids chemical constituents in water extract of Spatholobi Caulis. Some purification and analysis techniques like silica gel, D101-macroporous adsorptive resins, and Sephadex LH-20 column chromatographies as well as reversed phase high-performance liquid chromatography were used to isolate and analyze the phenolic acid esters and other type compounds from Spatholobi Caulis integrally. The structures of these compounds were identified by spectroscopic techniques such as nuclear magnetic resonance and high resolution mass spectrometries. Twenty-seven compounds, including phenolic acid, coumarin, lignan, terpene, alkaloid, and steroid compounds, were isolated from ethyl acetate and n-butanol fractions in water extract of Spatholobi Caulis, and they were identified as β-sitosterol(1), feruli acid methyl ester(2), syringaresinol(3),(+)-medioresinol(4),(+)-epipinoresinol(5), p-acetylphenol(6), bolusanthin Ⅳ(7), evofolin B(8), salicylic acid(9), trans-p-hydroxy-cinnamic acid(10), abscisic acid(11), m-hydroxyphenol(12), C-veratroylglycol(13), p-hydroquinone(14), 8,9-dihydroxymegastigma-4,6-dien-3-one(15), p-hydroxybenzoic acid(16), 6,9-dihydroxymegastigma-4,7-dien-3-one(17), protocatechuic acid(18), protocatechuic acid methyl ester(19), 5,7-dihydroxycoumarin(20), isolariciresinol(21), nicotinic acid(22), daucosterol(23),(+)-pinoresinol(24), stigmasterol(25), allantoin(26) and koaburaside(27), respectively. Furthermore, compounds 2-15, 19-22, 24 and 26 were isolated from genus Spatholobus for the first time.

Furanocoumarin A: A Novel Anticancer Agent on Human Lung Cancer A549 Cells from Fructus liquidambaris.

The fruit of Fructus liquidambaris, which is recently being used for cancer treatment, has a history to be used as a traditional medicine in China for thousands of years. Ten kg of dried F. liquidambaris was obtained with 70% alcohol-water solution under reflux for three times. The condensed extract was obtained from petroleum ether, ethyl acetate and N-butyl alcohol, respectively. Ethyl acetate extract was subjected to silica gel column, Sephadex LH-20, ODS column chromatography and RP-HPLC column chromatography to yield a new compound (1). The structure was identified through intensive analysis of NMR and MS spectra. The antitumor mechanism of the furanocoumarin A on human lung cancer A549 cells was confirmed by detecting the apoptosis-related proteins. Furanocoumarin A (1), a novel furanocoumarin constituent was isolated and identified from F. Liquidambaris. The IC50 value of furanocoumarin A on A549 cell lines was 65.28±5.36µM obtained by the method of MTT. The compound could induce the apoptosis of A549 cells by inducing 21.5% early apoptosis and 32.4% late apoptosis at the concentration of 60µmol/L. Western blot analysis indicated that protein expressions of p53, caspase 3 and Bax increased in a dose-dependent manner between the concentrations from 40 to 80µM. The protein expression of Bcl-2 decreased the concentration of 60 and 80µM. The ratio of Bcl-2 to Bax was inversely proportional to the dose concentration. Furanocoumarin A could be a novel anticancer agent from herbal medicine.

Isolation and characterization of some flavonoids from the leaf of Tapinanthus globiferus growing on Vitex doniana

Tapinanthus globiferus (A. Rich) Tiegh (Loranthacae) is a semi-parasitic plant growing on several plant species such as Vitex doniana. it is used in ethno-medicine for the treatment of fungal infection, itching, hypertension, ulcers, epilepsy, diabetes and cancer. The aim of this study was to isolate bioactive compound(s) from the leaf of T. globiferus. The powdered plant material was extracted with 90% methanol using maceration method and the resulting crude methanol leaf extract was partitioned into n-hexane, chloroform, ethylacetate and n-butanol fractions. The ethylacetate fraction was chromatographed on a silica gel and sephadex LH-20 column which led to the isolation of two flavonoids, and the identity of the compounds was determined on the basis of chemical test and NMR analysis. Based on the 1D and 2D NMR data, the compounds were 2-(3’4’-dihydroxyphenyl)-3,5,7-trihydroxy-4H-chromane-4-one (quercetin) and 2-(2,4-dihydroxyphenyl)-5,7-dihydroxy-3-(3-methylhexyl)-4H-chromen-4-one. This is the first report of isolation of these compounds from T. globiferus growing on Vitex doniana.

Constituents of flavonoids from Tridax procumbens L. and antioxidant activity

Background: To study the chemical constituents and antioxidant activities of flavonoids in Tridax procumbens L. Materials and Methods: Diaion HP-20, MCI-Gel CHP-20, Sephadex LH-20, and silica gel column chromatography were used to isolate and purify the chemical components of T. procumbens L., and the structures of the compounds were identified by physical and chemical properties and spectral techniques (1H-nuclear magnetic resonance [NMR] and 13C-NMR). The antioxidant activities of five flavonoids were evaluated by 2,2-diphenyl-1-picryl-hydrazyl method. Results: Six compounds were isolated and identified as luteolin-7-O-β-D-glucoside (1), 5,7,3'-trimethyl-4'-methoxyl-3-O-β-D-flavonoid glucoside (2), 8,3'-dihydroxyl-3,7,4'-trimethoxy-6-O-β-D-flavonoid glucoside (3), 4,2',4'-trihydroxy-6'-methoxyl-3'-isopentenyl chalcone (4), quercetin-7-O-β-D-glucopyranosyl-(2→1)-α-L-rhamnose (5), and nodakenin (6). Conclusions: All compounds except compound 3 were isolated from the plant for the first time. It was found that compounds 1–5 had significant antioxidant and free radical scavenging activities.

Chemical constituents from Saussurea pachyneura

Background: Genus Saussurea is famous for its use in traditional Chinese medicine. Objectives: Saussurea pachyneura, a plant from genus Saussurea, was studied for its secondary metabolites. Materials and Methods: The air-dried whole plant material was extracted by ethanol. The compounds from the ethanolic extract were isolated and purified by silica gel, octadecylsilyl-silica gel, and Sephadex LH-20 column chromatography. Their structures were identified based on the spectral analysis. Results: A total of 13 compounds, including a new long-chain fatty acid ester (4), were isolated and identified from the ethanolic extract of S. pachyneura. Conclusion: This is the first study to report the phytochemical analysis of S. pachyneura and also the first study to isolate the compounds from ethanolic extract of the whole plant material of S. pachyneura.

Free Radical Scavenging Principles of Salvia reuterana Boiss. Aerial Parts.

Salvia reuterana Boiss. is an aromatic perennial plant traditionally used for its anxiolytic and sedative properties. In the present study, various fractions and essential oil of S. reuterana aerial parts were investigated to find its free radical scavenging principles. Hydroalcoholic fraction with IC50 value of 112.6 ±3.2 μg mL-1 in DPPH assay demonstrated the highest free radical scavenging activity and was selected to further phytochemical investigation. RP-18 and Sephadex LH-20 column chromatography of the hydroalcoholic fraction resulted in the isolation and structural elucidation of four phenolic derivatives, including apigenin-7-O-β-D-glucopyranoside (1), luteolin-7-O-β-D-glucopyranoside (2), rosmarinic acid (3), and luteolin (4). Isolated compounds showed potent free radical scavenging activities (5.1-34.2 μg mL-1), compared with BHT (21.30 ± 1.9 μg mL-1). Twenty four compounds were also identified in GC-MS analysis of the plant essential oil, of which benzyl benzoate (26.64%), n-hexyl benzoate (22.99%) and n-hexyl isovalerate (6.04%) were the main compounds. The results of the present study introduced S. reuterana as a valuable source of natural phenolic antioxidants which can be utilized in prevention of oxidative stress related diseases. Moreover, interesting composition of S. reuterana essential oil, dominated by non-terpenes compounds (76.17%) especially aromatic derivatives, make it an appropriate candidate for more detailed studies.

Gastroprotective Effects of β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside from Bridelia ferruginea Stem Bark

The methanol extract of Bridelia ferruginea Benth. (Euphorbiaceae) stem bark (BFME) was partitioned in chloroform-methanol-water (2:2:1) mixture to obtain the Chloroform (CF) and Aqueous Methanol (AMF) fractions. The BFME, CF and AMF were screened for antiulcer activity using indomethacin-induced ulcer as activity guide. The CF provided the highest gastroprotection and was subsequently fractionated in a silica gel (60-200 mesh) column eluted with different mixtures of n-hexane and ethyl acetate (100:0; 95:5; 90:10; 80:20) to obtain six fractions (I-VI). Fractions III and VI offered the highest protection against indomethacin-induced ulcer and were further purified in a sephadex LH-20 column eluted with methanol to yield two compounds, BF1 and BF2. Using Nuclear Magnetic Resonance (1H-NMR, 13C-NMR) and electron impact mass spectroscopies, BF1 and BF2 were confirmed to be β-sitosterol and β-sitosterol-3-O-βD glucopyranoside respectively. The BFME, fractions, β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside elicited dose-related and significant (P<0.05) protection against various ulcers in rats. β-sitosterol, 100 and 300 mg/kg, produced 79.70, 82.18, 42.31, 44.87, 65.97, 70.83, 80.22 and 87.91% gastroprotection; while β-sitosterol-3-O-β-D-glucopyranoside, 100 and 300 mg/kg, caused 69.80 and 74.26, 33.33, 35.26, 84.03, 95.83, 83.52 and 85.71% gastroprotection against indomethacin-, ethanol-, cold restraint stress- and pylorus ligation- induced ulcers, respectively. Results demonstrated gastroprotective effects of B. ferruginea stem bark, attributable to β-sitosterol and β-sitosterol-3-O-β-D-glucopyranoside.

Barbaram, a bicyclic neolactam from the root and stem of Lycium barbarum

Barbaram (1), a neolactam together with other five known compunds (2–6) was isolated from the EtOAc-soluble fraction of an EtOH extract of the root and stem of Lycium barbarum by using silica gel, Sephadex LH-20 column chromatography and semi-preparative RP-C18 HPLC. Based on HR-TOF-MS, NMR spectral data, quantum chemistry ECD calculations and referencing to the data reported earlier, the structures of these compounds (1–6) were determined, specially including the absolute configuration of the neolactam (1). Compounds 2 and 4 showed significant anti-inflammatory activity in LPS-induced RAW 264.7 macrophages with the IC50 values of 8.98 ± 0.57 μM, 6.68 ± 0.77 μM.

A new guaiane-type sesquiterpenoid from Croton yunnanensis

Five sesquiterpenoids were isolated from 90% ethanol extract of Croton yunnanensis by silica gel,Sephadex LH-20 column chromatography,as well as prep-HPLC methods. Based on MS,1 D and 2 D NMR spectral analyses,the structures of the five compounds were identified as 11-methoxyl alismol(1),6β,7β-epoxy-4α-hydroxyguaian-10-ene(orientalol C,2),multisalactone D(3),arvestonol(4),and 4,5-dihydroblumenol A(5). Compound 1 was a new guaiane-type sesquiterpenoid. Compounds 2-4 were isolated from the Croton genus for the first time,and compound 5 was obtained from this plant for the first time.

A novel bacteriocin PE-ZYB1 produced by Pediococcus pentosaceus zy-B isolated from intestine of Mimachlamys nobilis: Purification, identification and its anti-listerial action

A novel bacteriocin, PE-ZYB1, was produced by Pediococcus pentosaceus zy-B that was isolated from the intestine of Mimachlamys nobilis and collected from the South China Sea. The bacteriocin PE-ZYB1 was purified using acetic ether extraction, Sephadex LH-20 column chromatography, sepharose cation exchange chromatography and reversed-phase high-performance liquid chromatography (RP-HPLC), in succession. Its molecular mass was determined to be MW 2019.22 Da, and its N-terminal amino acid sequence was DLFNKFNRSMQYGNGVE. The PE-ZYB1 was confirmed as a novel bacteriocin through a homology search, using UniProt BLAST (NO. A0A1Y0VLN8). Bacteriocin PE-ZYB1 was found to have extensive spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria. Bacteriocin PE-ZYB1 showed resistance to heat, was highly stable over pH 2–7 and was sensitive to trypsin. Increased extracellular electrical conductivity was detected for Listeria. monocytogenes treated by bacteriocin PE-ZYB1. Furthermore, bacteriocin PE-ZYB1 bactericidally destroyed the cell membrane integrity of L. monocytogenes, resulting in cell membrane perforation and permeabilization. These results indicate the potential of the bacteriocin PE-ZYB1 for use as a food bio-preservative.

Investigation on seco-prezizaane sesquiterpenes from fruits of Illicium lanceolatum and their neuroprotection activity

Ten seco-prezizaane sesquiterpenes were isolated from the water-soluble fraction of the fruit of Illicium lanceolatum using the combined methods of silica gel column chromatography,Sephadex LH-20 column chromatography,and RP-preparative HPLC. They were elucidated as majusanol E( 1),2α-hydroxycycloparviflorolide( 2),2β-hydroxy-3,6-dedioxypseudoanisatin( 3),majusanol A( 4),merrillianone( 5),cycloparvifloralone( 6),3α-hydroxycycloparvifloralone( 7),1,2-dehydrocycloparvifloralone( 8),henrylactone C( 9),and( 11) 7,14-ortholactone-3α-hydroxyfloridanolide( 10) according to the NMR data. All compounds were obtained from this plant for the first time. Neuroprotection activity,anti-Coxsackie B3 virus,and anti-H3 N2 virus experiments were carried out to test their bioactivities. The bioassay results showed that compounds 1,4,6,7,9 and 10 displayed weak protective effects of the damage of nerve SH-SY5 Y cell induced by monosodium glutamate.

A phytochemical study on Dichocarpum auriculatum, an endangered medicinal plant peculiar to China

ABSTRACT Objective Dichocarpum auriculatum, an endangered perennial herb, is endemic to China and has been used as folk medicines for the treatment of cough, hepatitis, scrofula, and epilepsy. However, there is no phytochemical report on this herbal so far. For the resource development and protective importance of this endangered medicinal plant, a phytochemical study was undertaken. Methods The chemical constituents were purified by silica gel column, Sephadex LH-20 column, and semi-preparative reversed phase HPLC. NMR and MS spectra were used for structural identification. Results Thirteen compounds were isolated from D. auriculatum. Their structures were characterized as jatrorrhizine (1), berberine (2), steponine (3), magnoflorine (4), coclauril (5), menisdaurin (6), menisdaurilide (7), aquilegiolide (8), (6R, 9S)-3-oxo-α-ionol-β-D-glucopyranoside (9), blumenol C glucoside (10), palmitic acid (11), dibutylphthalate (12), and auriculatum A (13). Conclusion Compound 13 is a new diester terephthalate derivative. All the compounds are obtained from the genus Dichocarpum for the first time, and compounds 9 and 10 have potential chemotaxonomic significance to the genus Dichocarpum.

Chemical constituents from heartwoods of Caesalpinia sappan with antiplatelet aggregation activities

ObjectiveTo clarify the active constituents of the heartwoods of Caesalpinia sappan, a traditional Chinese medicine with the functions of promoting blood circulation (Huoxue in Chinese) and removing blood stasis (Quyu in Chinese). MethodsThe chemical constituents were isolated and purified by combination of silica gel and Sephadex LH-20 column chromatography, along with semipreparative HPLC. Their chemical structures were established by multiple spectroscopic methods and comparison with literature data. The in vitro antiplatelet aggregation activities were evaluated using mouse platelet induced by AYPGKF-NH2, a gold agonist of protease-activated receptor 4 (PAR4). ResultsTwo new phenols, methyl 2-(4,4′,5′-trihydroxy-2′-(methoxymethyl) biphenyl-2-yloxy) acetate (1) and 1′-methylcaesalpin J (2), together with 24 known compounds (3–26), were isolated from the heartwoods of C. sappan. Among them, sappanchalcone (16) and brazilin (20) showed inhibitory activities against mouse platelet aggregation with IC50 values of 114.8 µmol/L and 100.8 µmol/L, respectively. ConclusionAntiplatelet compounds from C. sappan targeting at PAR4 are reported for the first time.

Sequences of Acretocins, Peptaibiotics Containing the Rare 1-Aminocyclopropanecarboxylic Acid, from Acremonium crotocinigenum CBS 217.70.

Sequences of Acretocins, Peptaibiotics Containing the Rare 1-Aminocyclopropanecarboxylic Acid, from Acremonium crotocinigenum CBS 217.70.

Chemical Profile and Antioxidant Activity of Zinnia elegans Jacq. Fractions.

Zinnia elegans (syn. Zinnia violacea) is a common ornamental plant of the Asteraceae family, widely cultivated for the impressive range of flower colors and persistent bloom. Given its uncomplicated cultivation and high adaptability to harsh landscape conditions, we investigated the potential use of Z. elegans as a source of valuable secondary metabolites. Preliminary classification of compounds found in a methanolic extract obtained from inflorescences of Z. elegans cv. Caroussel was accomplished using HR LC-MS techniques. The extract was then subjected to solid-phase extraction and separation using Sephadex LH-20 column chromatography, which resulted in several fractions further investigated for their antioxidant properties through lipoxygenase inhibition and metal chelating activity assays. Moreover, following additional purification procedures, structures of some active ingredients were established by NMR spectroscopy. The investigated fractions contained polyphenolic compounds such as chlorogenic acids and apigenin, kaempferol, and quercetin glycosides. Antioxidant assays showed that certain fractions exhibit moderate 15-LOX inhibition (Fr 2, IC50 = 18.98 μg/mL) and metal chelation (e.g., Fr 1-2, EC50 = 0.714–1.037 mg/mL) activities as compared to positive controls (20.25 μg/mL for kaempferol and 0.068 mg/mL for EDTA, respectively). For Fr 2, the 15-LOX inhibition activity seems to be related to the abundance of kaempferol glycosides. The NMR analyses revealed the presence of a kaempferol 3-O-glycoside, and a guanidine alkaloid previously not described in this species.

A new antioxidant flavonoid glycoside from flowers of Hosta plantaginea

Phytochemical investigation of the flowers of Hosta plantaginea led to isolate of one new flavonoid glycoside,plantanone C( 1) by silica gel,Sephadex LH-20,and RP-HPLC column chromatographies. Its structure was extensively determined on basis of HR-ESI-MS and NMR spectroscopic data. Compound 1 exhibited moderate antioxidant activity against DPPH radical scavenging activity,with an IC50 value of 240. 2 μmol·L~(-1).

Flavonoids from Chionanthus retusus (Oleaceae) Flowers and Their Protective Effects against Glutamate-Induced Cell Toxicity in HT22 Cells.

The dried flowers of Chionanthus retusus were extracted with 80% MeOH, and the concentrate was divided into EtOAc, n-BuOH, and H2O fractions. Repeated SiO2, octadecyl SiO2 (ODS), and Sephadex LH-20 column chromatography of the EtOAc fraction led to the isolation of four flavonols (1–4), three flavones (5–7), four flavanonols (8–11), and one flavanone (12), which were identified based on extensive analysis of various spectroscopic data. Flavonoids 4–6 and 8–11 were isolated from the flowers of C. retusus for the first time in this study. Flavonoids 1, 2, 5, 6, 8, and 10–12 significantly inhibited NO production in RAW 264.7 cells stimulated by lipopolysaccharide (LPS) and glutamate-induced cell toxicity and effectively increased HO-1 protein expression in mouse hippocampal HT22 cells. Flavonoids with significant neuroprotective activity were also found to recover oxidative-stress-induced cell damage by increasing HO-1 protein expression. This article demonstrates that flavonoids from C. retusus flowers have significant potential as therapeutic materials in inflammation and neurodisease.

Chemical compositions from roots of Erythrina corallodendron

This project is to investigate chemical compositions from the roots of Erythrina corallodendron. Through the methods of silica gel,ODS,Sephadex LH-20 column chromatography and preparative HPLC,15 compounds were isolated from the 95% ethanol extract of the roots of E. corallodendron. Based on spectroscopic techniques,the structures of these compounds were identified as 10,11-dioxoerythraline( 1),erythrinine( 2),erythraline( 3),11-methoxyerythraline( 4),cristanines B( 5),erythratine( 6),erysotrine( 7),medioresinol( 8),( ±)-ficusesquilignan A( 9),( +)-pinoresinol( 10),nicotinic acid( 11),dibutyl phthalate( 12),vanillic acid( 13),3-hydroxy-1-( 4-hydroxy-3-methoxyphenyl)-1-propanone( 14),and syringic acid( 15). Compounds 8-10 are isolated from genus Erythrina for the first time and all compounds are isolated from E. corallodendron for the first time. Furthermore,this paper screened the antioxidant and cytotoxic activities of the compounds using models of liver microsomal oxidation inhibition and MTT.

Antidiabetic Activities of Korean Red Pine (Pinus densiflora) Inner Bark Extracts

This study was carried out to investigate the potential of Korean red pine (Pinus densiflora) inner bark extracts as an antidiabetic agent. The ethyl acetate soluble fraction of the bark extracts was chromatographed on a Sephadex LH-20 column to yield five compounds, which structures were elucidated by NMR spectroscopy. The isolated compounds were (+)-catehin, (−)-epicatechin, taxifolin, taxifolin-3′-O-β-D-(+)-glucose and ñ-courmaric acid. The antidiabetic activity of the different fractions, including the crude extracts and isolated compounds, was evaluated by β-cells insulin secretion and glucose uptake in skeletal muscle cells. The insulin secretion was 128% for taxifolin at 25 μg/mL. However, the other samples had no effect on this test. For the glucose uptake activity assay, 1 μM insulin and 2 mM metformin were used as controls. Both the crude extract and taxifolin showed relatively low activity values, but the other samples yielded glucose uptake values over 260%. ρ-courmaric acid showed the highest uptake (270%). The results confirmed that Korean red pine extracts may be used as a hypoglycemic agent.

6-(2-Formyl-5-methyl-1H-pyrrol-1-yl)hexanoic Acid as a Novel Retronasal Burnt Aroma Compound in Soy Sauce Aroma-Type Chinese Baijiu.

Two-step high-performance liquid chromatography (HPLC) separation in combination with taste dilution analysis was used to screen the retronasal burnt aroma compounds in soy sauce aroma-type Chinese Baijiu (Chinese traditional liquor). A major retronasal burnt aroma region was purified by Sephadex LH-20 column chromatography and HPLC separation from the soy sauce aroma-type Baijiu sample. A novel retronasal burnt aroma compound was identified as 6-(2-formyl-5-methyl-1H-pyrrol-1-yl)hexanoic acid (1) by means of high-resolution mass spectrometry and 1D/2D nuclear magnetic resonance. The retronasal burnt aroma threshold of compound 1 was determined to be 209.5 μg/L in 53% ethanol aqueous solution. Quantitative analysis showed that the concentration of compound 1 ranged from below the limit of quantitation (3.8 μg/L) to 224.2 μg/L in the Baijiu samples. Moreover, the sensory evaluation also revealed that compound 1 may contribute to the retronasal burnt aroma of the Baijiu at its subthreshold concentration by the additive effect with the recently reported retronasal burnt aroma compound 2-hydroxymethyl-3,6-diethyl-5-methylpyrazine.