ODS Column Chromatography Research Articles

Triterpenoids from Ganoderma lucidum and their cytotoxic activities

From the ethyl acetate fraction of the fruiting body of Ganoderma lucidum, a new triterpenoid, ethyl 7β-hydroxy-4,4,14α-trimethyl-3,11,15-trioxo-5α-chol-8-en-24-oate (4), named ethyl lucidenates A, along with three known compounds, ganodermanondiol (1), lucidumol B (2) and methyl lucidenates A (3) were isolated by silica gel column, ODS column chromatography and PHPLC. Their structures were established on the basis of spectroscopic analysis and chemical evidence. The isolated compounds were tested using in vitro MTT assay for their cytotoxic activities against the K562, HL-60, CA46, HepG2, SW480 and SMMC-7221 cancer cell lines. Among them, compound 4 showed cytotoxicity against HL-60 and CA46 cancer cell lines with IC50 values of 25.98 and 20.42 µg mL−1, respectively.

A new feruloyl tyramine glycoside from the roots of Achyranthes bidentata

Aim To study the chemical constituents of the roots of Achyranthes bidentata Bl. Methods The chemical constituents were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatographies and preparetive HPLC. Their structures were elucidated on the basis of 1D and 2D NMR analyses. Results Two feruloyl tyramine glycosides and seven triterpenoid saponins were obtained and identified as N-trans-feruloyl-3-methoxytyramine-4′- O-β-D-glucopyranoside ( 1), N-trans-feruloyl-3-methoxytyra mine-4- O-β-D-glucopyranoside ( 2), PJS-1 ( 3), chikusetsusaponin IVa ( 4), oleanolic acid 3- O-[β-D-glucuronopy ranoside-6- O-methyl ester]-28- O-β-D-glucopyranoside ( 5), oleanolic acid 3- O-[β-D-glucuronopyran-oside-6- O-ethylester]-28- O-β-D-glucopyranoside ( 6), oleanolic acid 3- O-[β-D-glucuronopyranoside-6- O-butyl ester]-28- O-β-D-glucopyranoside ( 7), ginsenoside R 0 ( 8) and hederagenin-28- O-β-D-glucopyranosyl ester ( 9). Conclusion Compound 1 is a new feruloyl tyramine glycoside, while compounds 2 and 9 are reported from A. bidentata for the first time.

A new phenylpropanoid glycoside from the fruits of Illicium simonsii

Aim To study the constituents and bioactivity of Illicium simonsii. Methods The extracts of the fruits of Illicium simonsii were submitted to a combination of chromatographic materials, silica gel, ODS column chromatography and finally preparative HPLC to obtain eight compounds which were further evaluated for their cytotoxicity by MTT method. Results A new phenylpropanoid glycolside, 2, 4-dihydroxy-allylbenzene-2- O-β-D-glucopyranoside ( 1), together with seven characteristic sesquiterpene lactones, oligandrumin B ( 2), oligandrumin D ( 3), anisatin ( 4), veranisatin D ( 5), pseudomajucin ( 6), 1α-hydroxy-3-deoxy-pseudoanisatin ( 7), 8α-hydroxy-10-deoxycyclomerrillianolide ( 8) were isolated. Conclusion Compound 1 is new. Compounds 1, 2, 3, 5–8 were isolated from this plant for the first time. None of the isolates showed inhibitory effects on the growth of non-small cell lung tumor cell line A549.

A New Phragmalin-type Limonoid from Chukrasia tabularis var. velutina

Aim To study the limonoid constituents of Chukrasia tabularis var. velutina. Methods The constituents were isolated with silica gel, ODS column chromatography and preparative HPLC, their structures were elucidated on the basis of the NMR and HRESIMS spectral data. Results Two phragmalin-type limonoids, tabulalin F ( 1) and tabulalin ( 2) were isolated. Conclusion Tabulalin F ( 1) was a new compound and tabulalin ( 2) was isolated from this plant for the first time.

Dinapinones, novel inhibitors of triacylglycerol synthesis in mammalian cells, produced by Penicillium pinophilum FKI-3864

Dinapinone A, a novel biaryl dihydronaphthopyranone, was isolated from the culture broth of Penicillium pinophilum FKI-3864 by solvent extraction, silica gel and ODS column chromatography and HPLC. Dinapinone A showed very potent inhibition of triacylglycerol (TG) synthesis in intact Chinese hamster ovary K1 (CHO-K1) cells with an IC(50) value of 0.097 μM. Dinapinone A was found to be a mixture of stereoisomers, resulting in its separation into dinapinones A1 and A2 by HPLC using a C30 reverse-phase column. Dinapinone A1 did not inhibit TG synthesis in CHO-K1 cells even at 12 μM, and dinapinone A2 showed less potent inhibition (IC(50); 0.65 μM) than dinapinone A; however, a mixture of isolated dinapinones A1 and A2 (a 1:1 ratio) recovered the potent TG inhibitory activity (IC(50); 0.054 μM). A similar effect of dinapinone on TG synthesis in intact Raji cells was also observed.

Isolation of megastigmane sesquiterpenes from the silkworm (Bombyx mori L.) droppings and their promotion activity on HO-1 and SIRT1

The silkworm (Bombyx mori L.) droppings were extracted with 80% aqueous MeOH, and the concentrated extract was partitioned in succession with EtOAc, n-BuOH, and H(2)O. From the EtOAc fraction, five megastigmane sesquiterpenes were isolated through repeated silica gel and ODS column chromatography. According to the results of spectroscopic data, such as NMR, MS, and IR, the chemical structures of the isolated compounds were determined as (3S,5R,8R)-3,5-dihydroxymegastigma-6,7-dien-9-one (1), (S)-dehydrovomifoliol (2), (6R,7E,9R) -9-hydroxy-4,7-megastigmadien-3-one (3), (3S,5R,6S,7E)-3,5,6-trihydroxy-7-megastigmen-9-one (4), (6R,9R)-9-hydroxy-4-megastigmen-3-one (5). Compounds 2 through 5 were isolated for the first time from silkworm droppings. GC/MS analysis indicated silkworm powder contained compound 3, and mulberry leaves contained compound 4. Compounds 1 and 5 increased the expression of heme oxygenase-1 and SIRT1 in HepG2 and HEK239 cells, respectively. Heme oxygenase-1 is considered to be an antioxidant enzyme that catabolizes heme to carbon monoxide, free iron and biliverdin, while SIRT1 is the mammalian homologue of the yeast silent information regulator (Sir)-2, which are involved in the suppression of inflammatory mediators or factors that may be used to improve atopy-related symptoms.

Determination of Pindone in Animal Products, Fishery Products, and Honey by LC-MS/MS

A sensitive and selective analytical method for the determination of the rodenticide pindone in animal products, fishery products, and honey by LC-MS/MS was developed. Pindone was extracted with acidified acetone, and the crude extract was purified by liquid-liquid partitioning, followed by silica gel and ODS column chromatography. LC separation was performed on an ODS column with methanol/water containing ammonium acetate as the mobile phase, and detection was carried out using tandem mass spectrometry (MS/MS) with electrospray ionization (ESI) in the negative mode. The average recoveries from fortified bovine muscle, bovine liver, bovine fat, chicken muscle, salmon, eel, freshwater clam, egg, milk, and honey spiked at 0.001 mg/kg were in the range of 76-92%, and the relative standard deviations were 4-8%. The limit of quantitation (S/N≥10) of the developed method was 0.001 mg/kg for all the tested foods.

Anthraquinone Glycosides from the Roots of Prismatomeris connata

Aim To study the anthraquinone glycosides and their cytotoxicity from the roots of Prismatomeris connata. Methods The chemical constituents were isolated by Diaion HP-20 resin, silica gel, Sephadex LH-20 and ODS column chromatography and the structures were elucidated on the basis of spectral analysis. The cytotoxic activity was evaluated by MTT method. Results Six known anthraquinone glycosides were obtained and their structures were identified as 1- O-methylrubiadin 3- O-β-primeveroside ( 1), damnacanthol 3- O-β-primeveroside ( 2), rubiadin 3- O-β-primerveroside ( 3), lucidin 3- O-β-primeveroside ( 4), 1, 3-dihydroxy-2-(methoxymethyl) anthraquinone 3- O-β-primerveroside ( 5), and digiferruginol ω-gentiobiose ( 6). Conclusion All the compounds were isolated from this genus for the first time.

Five Lignans and an Iridoid from Sambucus williamsii

Aim To study the chemical constituents of the 60% ethanol extract of the stems of Sambucus williamsii Hance. Methods Compounds were isolated and purified by macroporous adsorptive resin D101, silica gel, and ODS column chromatographies and preparative HPLC. Their structures were identified by spectroscopic methods. Results Five lignans and one iridoid were isolated from the stems of Sambucus williamsii Hance. and their structures were identified as dehydrodiconiferyl alcohol-4- O-β-D-glucopyranoside ( 1), dihydrodehydrodiconiferyl alcohol-4- O-β-D-glucopyranoside ( 2), (7 S, 8 R)-dihydro-3'-hydroxy- 8-hydroxy-methyl-7-(4-hydroxy-3-methoxy-phenyl)-1'-benzofuranpropanol ( 3), erythro-1-(4-hydroxy-3-methoxyphenyl)-2-[2-hydroxy-4-(3-hydroxypropyl) phenoxy]-1, 3-propanediol ( 4), threo-1-(4-hydroxy-3-methoxyphenyl)-2-[2-hydroxy-4-(3-hydroxypropyl) phenoxy]-1, 3-propanediol ( 5), and morroniside ( 6) with about 70% α-morroniside and 30% β-morroniside. Conclusion Compounds 1, 4 and 5 were isolated from this genus for the first time, and compounds 2, 3 and 6 were obtained from the plant for the first time.

적콜라비 (Brassica oleracea var. gongylodes) 새싹으로부터 sterol 화합물의 분리 및 동정

The sprouts of Brassica oleracea var. gongylodes were extracted with 100% MeOH and the concentrated extract was partitioned with EtOAc, n-BuOH and , successively. From the EtOAc fraction, three sterols were isolated through the repeated silica gel and ODS column chromatographies. On the basis of physico-chemical and spectroscopic data including NMR, MS, and IR, the chemical structures of the sterols were determined as -sitosterol (1), brassicasterol (2), and 7-ketobrassicasterol (3). Compound 1 is usually observed in plant. Compounds 2 is observed in Brassica sp., and compounds 3 have very rarely occurred in natural source including plant.

ChemInform Abstract: Roselipins, Inhibitors of Diacylglycerol Acyltransferase, Produced by Gliocladium roseum KF-1040.

Gliocladium roseum KF-1040, a marine isolate, was found to produce a series of new inhibitors of diacylglycerol acyltransferase (DGAT). Four active compounds, designated roselipins 1A, 1B, 2A and 2B, were isolated from the fermentation broth of the producing strain by solvent extraction, ODS column chromatography and preparative HPLC. The highest production of roselipins was observed when cultured in the medium containing natural sea water. Roselipins inhibit DGAT activity with IC50 values of 15 approximately 22 microM in an enzyme assay system using rat liver microsomes.

ChemInform Abstract: Phenochalasins, Inhibitors of Lipid Droplet Formation in Mouse Macrophages, Produced by Phomopsis sp. FT-0211.

Phomopsis sp. FT-0211, a soil isolate, was found to produce inhibitors of lipid droplet formation in mouse peritoneal macrophages. Structurally related new compounds designated phenochalasins A and B were isolated from the fermentation broth of the producing strain by solvent extraction, ODS column chromatography and preparative HPLC. Phenochalasin A caused a dose-dependent reduction in the number and size of lipid droplets in macrophages without any cytotoxic effect at least up to 20 microm. On the other hand, phenochalasin B showed inhibition of lipid droplet formation with a severe cytotoxic effect on macrophages.

Chemical constituents of Acacia catechu

To study the chemical constituents of Chinese medicine Acacia catechu. Isolation and purification were carried out on normal phase silica gel, Sephadex LH-20, ODS column chromatography etc. Constituents were identified by physicochemical properties and spectral analysis. Twelve compounds were identified as 4-hydroxybenzoic acid( 1), kaempferol (2), quercetin (3), 3,4',7-trihydroxyl-3', 5-dimethoxyflavone (4), catechin (5), epicatechin (6), afzelechin (7), epiafzelechin (8), mesquitol(9), ophioglonin (10), aromadendrin (11), and phenol (12). Compounds 7, 12 were isolated from A. catechu for the first time, and compounds 4, 9-11 were isolated from the genus Acacia for the first time.

Chemical constituents from stem barks of Vernonia cumingiana

The chemical constituents from the stem barks of Vernonia cumingiana were investigated. Various chromatographic techniques such as silica gel chromatography, Sephadex LH-20, ODS column chromatography and HPLC were used to isolate and purify the constituents. The structures were elucidated by spectral methods. Twelve compounds were isolated from the 95% ethanol extract and their structures were elucidated as methyl 3,5-dicaffeoylquinate (1), methyl 3,4-dicaffeoylquinate (2), ethyl 3,4-dicaffeoylquinate (3), methyl 3,4,5-tricaffeoylquinate (4), stigmasterol (5), alpha-spinasterol (6), beta-sitosterol (7), 24-methylene-lanosta-9 (11)-en-3beta-acetate (8), ethyl gallate (9), di-n-butyl-phthalate (10), stearic acid (11) and palmitic acid (12). Compounds 1-12 were isolated from this plant for the first time.

Isolation of Salvianolic acid A, a Minor Phenolic Carboxylic Acid of Salvia miltiorrhiza

Salvianolic acid A (Sal A), a caffeic acid derivative present in Salvia miltiorrhiza (Danshen), has shown significant biological activities, some of which are more potent than those of salvianolic acid (Sal B), the most abundant and bioactive compound in Danshen. However, its low content in the raw material makes it difficult to be used in the clinic. In this study, a novel procedure was developed to isolate Sal A from Salvia miltiorrhiza. Using this procedure, Sal A could be enriched to 1.7% in the raw material by hydrolyzing Sal B. The solution was further purified by macroporous resin and ODS column chromatography. The yield of Sal A was 0.5% with a purity of 98.0%. The procedure is efficient and convenient and is considered suitable for the separation of Sal A from Danshen.

Nosokomycins, new antibiotics discovered in an in vivo-mimic infection model using silkworm larvae. I: Fermentation, isolation and biological properties

The in vivo-mimic assay system using silkworm larvae was used as a screening tool to discover antibiotics against methicillin-resistant Staphylococcus aureus (MRSA). Microbial culture broths were screened in this in vivo-mimic assay system and a culture broth of Streptomyces sp. K04-0144 was selected. New antibiotics, designated nosokomycins A-D, were isolated from the culture broth by HP-20 and ODS column chromatography and HPLC. Nosokomycins inhibited the growth of MRSA with MIC values of 0.125 microg ml(-1) using the liquid microdilution method. Furthermore, MRSA-infected silkworms survived when nosokomycin A or B was injected at a dose of 50 microg per larva.

Inhibition on LDL-oxidation by Phenolic Compounds from the Fruit Body of Phellinus linteus

The fruit body of Phellinus linteus was extracted with 80% aqueous MeOH, and the concentrated extract was successively partitioned using EtOAc, n-BuOH and . From the EtOAc and n-BuOH fractions, three phenolic compounds were isolated through repeated silica gel and ODS column chromatography. The chemical structures of these compounds were determined as 2-(3`,4`-dihydroxyphenyl)-1,3-benzodioxole-5-aldehyde (1), 4-(3,4-dihydroxyphenyl)-3-buten-2-one (2), and protocatechuic acid methyl ester (3) by spectroscopic data including NMR, MS and IR. Compounds exhibited low density lipoprotein (LDL) antioxidant activity with values of 1.7, 0.7, and , respectively.

Isolation and Identification of Lipids from the Silkworm (Bombyx mori) Droppings

Silkworm (Bombyx mori) droppings were extracted with 80% aqueous MeOH, and the concentrated extract was partitioned successively with EtOAc, n-BuOH, and H2O. Three lipids were isolated from the EtOAc fraction by repeated silica gel and ODS column chromatography. Physico-chemical data including NMR, MS, and IR indicated that the isolated lipids were β-sitosterol (1), corchorifatty acid E methyl ester (2), and (8E)-10-hydroxydec-8-enoic acid methyl ester (3). Compounds 2 and 3 have, prior to this study, not previously been documented in nature.

Isolation of Triterpene Glycosides from the Fruit of Cornus kousa

Dried, unripe fruits of Cornus kousa Burg, were extracted with 80% aqueous MeOH and the concentrated extracts were partitioned using EtOAc, n-BuOH and H2O, successively. From the EtOAc fraction, three triterpene glycosides were isolated through repeated SiO2 and ODS column chromatography. Based on NMR, MS and IR spectroscopic data, the chemical structures of the compounds were determined to be tormentoside (1), niga-ichigoside F2 (2), and arjunglucoside (3). This is the first report of these compounds being isolated from this plant.

HPLC analysis, semi-preparative HPLC preparation and identification of three impurities in salidroside bulk drug

Salidroside is a bioactive compound mainly distributed in Rhodiola L. (Crassulaceae). It has been widely used in Chinese traditional medicine. In this paper, three impurities were found during the analysis of salidroside bulk drug. The enrichment of impurities was carried out by ODS column chromatography, using methanol–water (13:87, v/v) as eluent and the purification of impurities was achieved by semi-preparative HPLC, using methanol–water (11:89, v/v) as mobile phase, respectively. Three impurities were characterized as 4-(2-hydroxylethyl)-phenol-1-O-β-d-glucopyranoside, 4-hydroxyphenacyl-d-glucopyranoside and p-acetylphenyl-O-β-d-glucopyranoside by a variety of spectral data (IR, UV, MS, 1H NMR, 13C NMR, DEPT and 2D NMR). The simultaneous quantitative determination of salidroside and its impurities (Imp. 1, 2 and 3) was performed by reverse-phase HPLC method with UV detection. Specificity, linearity, sensitivity, precision and accuracy were evaluated.