Untersuchungen zur Anreicherung von Ergosterol und Ubichinonen aus Lipid-Kohlenwasserstoff-Fraktionen

Adsorption on Silicagel was used for the enrichment of ergosterol and ubiquinone-9 from a lipid-hydrocarbon extract. The main components of the lipid-hydrocarbon extract were hydrocarbons of gas oil (B. p. 513 to 653 K)phosphatides, glycerides, and fatty acids. By adsorption on silicagel (1 part silicagel to 2–3 parts of lipid-hydrocarbon extract dissolved in hexane) were ergosterol and ubiquinone-9 enriched and nearly completely separarated from hydrocarbons and phosphatides, partly from glycerides and fatty acids. Ergosterol and ubiquinone-9 can be obtained by a simplified separation from the enriched fraction.     

Untersuchungen zur Anreicherung von Ubichinon-9 aus Lipid-Kohlenwasserstoff-Fraktionen mittels Kurzwegdestillation

Short path distillation was used for the enrichment of ubiquinone-9 from a lipid hydrocarbon extract. The main components of the lipid hydrocarbon extract were hydrocarbons of gas oil (b. p. 513 to 653 K), phosphatides, glycerides, and fatty acids. Phosphatides were isolated by extraction with acetone before distillation. The acetone soluble fraction was distilled by a pressure of ≦ 0.5 kPa and a temperature of 433 to 453 K. The concentration of the ubiquinone-9 in the residue of distillation was three times higher than in the acetone soluble fraction. Ubiquinone-9 can be obtained by a simplified separation from the enriched fraction.     

Untersuchungen zur chemischen Zusammensetzung der Lipidfraktion von Acinetobacter calcoaceticus

Lipids were extracted from the cells of Acinetobacter calcoaceticus EB 10 C IMET B 395 grown on gas oil (Bp. 240–360 °C) with benzine/alcohol (80 : 20). The lipid-hydrocarbon-extract obtained by this extraction method was 18.4%. The extract was composed of hydrocarbons, waxes, phospholipids, fatty acids, glycerides, and ubiquinones. The main components among the lipids were waxes. The compositions of phospholipid, fatty acid, wax, and ubiquinone fractions were analysed.     

Disulphates of 16-oxygenated ketonic c19 steroids as biliary metabolites of androsterone sulphate in female rats

The chemical synthesis of 16β-hydroxyandrosterone was described preparatory to studies of the disulphates of the 16-oxygenated ketonic C₁₉ steroids present in the bile of female rats dosed with [³H]androsterone sulphate. The biliary metabolites were separated by chromatography on Sephadex LH-20 to afford monosulphate and dicon jugate fractions. After solvolysis of the diconjugate fraction, the liberated steroids were separated by partition chromatography on Celite 545 and analyzed by gas chromatography-mass spectrometry. In addition to 3α, 17β-dihydroxy-5α-androstan-16-one isolated previously, 16β-hydroxyandrosterone was identified as a disulphate.     

Mass spectrometric analysis of cytokinins in plant tissues : v. Identification of the cytokinin complex of datura innoxia crown gall tissue

The cytokinin complex of Datura innoxia Mill. crown gall tissue was purified by ion exchange, Sephadex LH-20 chromatography and reversed-phase high performance liquid chromatography. By gas chromatography-mass spectrometry using ²H-labeled compounds, the following cytokinins were identified in the basic fraction eluting from a cation exchange column: zeatin, zeatin riboside, dihydrozeatin, dihydrozeatin riboside, their corresponding O-glucosides, 7- and 9-glucosides of zeatin, 9-glucoside of dihydrozeatin, isopentenyladenine, and isopentenyladenosine. Zeatin riboside 5′-monophosphate was the major cytokinin nucleotide in the tissue. In addition, dihydrozeatin riboside and isopentenyladenosine were identified in the nucleotide fraction following enymic degradation.     

Inhibition of nitric oxide production in LPS-stimulated RAW 264.7 cells by stem bark of Ulmus pumila L.

This study was designed to isolate and identify a potent inhibitory compound against nitric oxide (NO) production from the stem bark of Ulmus pumila L. Ethyl acetate fraction of hot water extract registered a higher level of total phenolics (756.93 mg GAE/g) and also showed strong DPPH (IC₅₀ at 5.6 μg/mL) and ABTS (TEAC value 0.9703) radical scavenging activities than other fractions. Crude extract and its fractions significantly decreased nitrite accumulation in LPS-stimulated RAW 264.7 cells indicating that they potentially inhibited the NO production in a concentration dependent manner. Based on higher inhibitory activity, the ethyl acetate fraction was subjected to Sephadex LH-20 column chromatography and yielded seven fractions and all these fractions registered appreciable levels of inhibitory activity on NO production. The most effective fraction F1 was further purified and subjected to ¹H, ¹³C-NMR and mass spectrometry analysis and the compound was identified as icariside E₄. The results suggest that the U. pumila extract and the isolated compound icariside E₄ effectively inhibited the NO production and may be useful in preventing inflammatory diseases mediated by excessive production of NO.     

25-OHD3-26,23-lactone: a metabolite of vitamin D3 that is 5 times more potent than 25-OHD3 in the rat plasma competitive protein binding radioassay.

A new metabolite of Vitamin D₃ (25-OHD₃-26,23-lactone) has been found in the plasma of Vitamin D₃-toxic pigs and cows. This metabolite is at least 5 times more potent than 25-OHD₃ in the displacement of [³H]-25-OHD₃ from rat plasma protein binding sites under short-term incubation. This metabolite co-migrates with 24,25-(OH)₂D₃ on Sephadex LH-20 columns developed in chloroform:hexane 65:35 and with 25,26-(OH)₂D₃ on Sephadex LH-20 columns developed in hexane:chloroform:methanol 9:1:1. The presence of 25-OHD₃-26,23-lactone represents a possible contaiminant in the assay of 24,25-(OH)₂D₃ or 25,26-(OH)₂D₃ if only Sephadex LH-20 is used for pre-assay purification. 25-OHD₃-26,23-lactone is, however, resolved from 24,25-(OH)₂D₃ by high pressure liquid chromatography (HPLC) using Zorbax Sil silicic acid columns developed in either isopropanol:hexane 8:92 or isopropanol:methylene chloride 2.5:96.5. We assayed for the presence of this new metabolite of Vitamin D₃ and found it to be present in normal pig plasma and undetectable in normal cow plasma. Concentrations were elevated to 10–20 ng/ml following massive injection of Vitamin D₃ to both species.     

Identification of 5-hydroxytryptamine binding substances from rat brain stem

Abstract— Godwin & Sneddon (1975) reported the binding of 5-hydroxy-[³H]tryptamine (5-HT) on a Sephadex LH-20 column to‘proteolipid material’extracted with n-butanol from rat brain stem. An examination of this‘proteolipid material’with TLC showed the main constituents to be cerebroside sulfate (CS), monophosphoinositide (PI), and diphosphoinositide. The elution profiles of [³H]5-HT incubated with purified CS or with a mixture of CS and PI were similar to that of the brain extract on the same column. Because the elution profile of the mixture of CS and PI was more similar to that of the brain extract, it was concluded that what was suggested to be a possible proteolipid‘5-HT receptor’was mainly two acidic lipids. The elution profile of [³H]5-HT incubated with purified PI, however, was similar to [³H]5-HT eluted alone. This suggested that either PI did not bind to 5-HT or that the PI-5-HT complex possesses different Chromatographie behavior than PI. To test this latter possibility, [¹⁴C]5-HT and [³H]PI were incubated then eluted on a Sephadex LH-20 column with a continuous gradient of increasing polarity. The gradient first eluted PI, then an apparent PI-5-HT complex, and finally 5-HT. This demonstrated that PI will bind to 5-HT on a Sephadex LH-20 column and that the PI-5-HT complex is probably more polar than PI.     

Does estradiol stimulate in vivo production of 1,25-dihydroxyvitamin D3 in the rat?

Female rats were injected with estradiol benzoate (3 mg/kg daily) or corn oil for 8 days before receiving 325 picomoles of 25-[26,27³H]-hydroxyvitamin D₃ intravenously. Eighteen hours later lipids were extracted from plasma, gut mucosa and kidneys and vitamin D₃ metabolites were separated using Sephadex LH-20 and chloroform: hexanes (65:35) column chromatography. Estradiol treatment increased 1,25-[26,27-³H]-dihydroxyvitamin D₃ content in all three isssues.     

Effect of water on hydrolysis of olive oil by immobilized lipase in reverse phase system

Summary Candida rugosa lipase immobilized by adsorption on the swollen Sephadex LH-20 and Sephadex LH-60 could effectively hydrolyze olive oil at high concentration in a reverse phase system. Initial water content was found to be the most important factor that determines both the hydrolysis rate and the degree of hydrolysis; approximately 54% of water in the gels was readily utilized.     

Gas Chromatography-Flame Thermionic Detector Analysis of Cytokinins in Etiolated Squash Seedlings using 14C-BA as an Internal Standard

Cytokinin contents in cotyledon, hypocotyl and root of etiolatedsquash (Cucurbita maxima Duch.) seedlings were determined byinstrumental analysis using ¹⁴C-benzyladenine (¹⁴C-BA) as aninternal standard. Crude extracts were purified using insolublepolyvinylpyrrolidone, cellulose-phosphate column and SEP-PAKC18 cartridge, then applied to a Sephadex LH-20 column to separatezeatin riboside (ZR), isopentenyl adenosine, isopentenyl adenine,¹⁴C-BA and a mixture of zeatin (Z) and dihydrozeatin (DHZ).The recovery rate for the cytokinin fractions after LH-20 wascorrected by ¹⁴C-BA. Each cytokinin fraction was further purifiedby HPLC which also separated Z and DHZ in the LH-20 fraction.Before permethylation, ¹⁴C-BA was added to each of the cytokininfractions to correct the methylation rates. Each methylatedcytokinin fraction was again purified by HPLC, then subjectedto gas chromatography with a capillary column and flame thermionicdetector. The detection limit of cytokinins by this system was0.1 ng. cis-ZK was the most abundant cytokinin in all tissues of theetiolated squash seedlings. Active cytokinins such as trans-ZRand trans-Z were mostly found in cotyledons with lesser amountsin the roots. DHZ was most abundant in the cotyledon. All cytokininsisolated by this procedure were confirmed by gas chromatographyselectedion monitoring. (Received December 26, 1986; Accepted June 1, 1987)     

Occurrence of Indole-3-Acetic Acid in Buds of Pinus silvestris

The major ether-soluble, growth-stimulating substance detected by the Avena coleoptile straight-growth test in extract from sprouting buds of Scots pine (Pinus silvestris L.) was identified as indole-3-acetic acid by Rf values in 5 solvent systems and by its elution volume in ethanol on a Sephadex LH-20 column. When the substance was applied to the growth solution of wheat roots in a special test the growth in length of the roots was at first inhibited, but growth was recovered after about 6 hours in the same manner as when small quantities of IAA were applied. The extracts also contained large amounts of growth inhibitors which interfered with the auxin response if they were not removed.     

Cytokinins in developing fruits of Moringa pterigosperma Gaertn.

The existence of cytokinins both as a free form and as a constituent of t-RNA was investigated in young fruits of Moringa pterigosperma Gaertn. Purified methanol extract was separated into butanol insoluble and butanol soluble fractions. The cytokinin(s) in the butanol insoluble fraction was tentatively identified as zeatin nucleotide. The butanol soluble fraction contained cytokinins and was chromatographed on Sephadex LH-20 with 35% ethanol. The two active fractions from LH-20 column coincided with zeatin and zeatin riboside. Cytokinin per g tissue was high in early stages of fruit growth and then remained more or less constant. Alkaline phosphatase hydrolysis of t-RNA hydrolysate of fruit tissue showed considerable cytokinin activity.     

Analysis of metabolic profiles of prostaglandins in urine using a lipophilic anion exchanger

A method is described for the fractionation of prostaglandins and their metabolites in urine. Following acidification and extraction on Amberlite XAD-2, samples were separated by chromatography on the lipophilic anion exchanger diethyl-aminohydroxypropyl Sephadex LH-20 into fractions containing neutral compounds, monocarboxylic, dicarboxylic and polycarboxylic acids. The compounds in resulting fractions were further separated by reversed phase partition chromatography. As an application, the metabolic profiles in urine of [9β-^3H]-labeled prostaglandin F_2α¹ and prostaglandin analogs 15-methyl-PGF_2α and 16,16-dimethyl-PGF_2α were investigated in the cynomolgus monkey. It was demonstrated that the resolution of individual prostaglandin metabolites by reversed phase partition chromatography was considerably simplified by initial group separation on the anion exchanger, and several metabolites were much purified. A glucuronic acid conjugate of the main metabolite of 15-methyl-PGF_2α (dinor-15-methyl-PGF_2α) was tentatively identified using computerized gas chromatography - mass spectrometry.     

Microbial Degradation of Alkyl Carbazoles in Norman Wells Crude Oil

Norman Wells crude oil was fractionated by sequential alumina and silicic acid column chromatography methods. The resulting nitrogen-rich fraction was analyzed by gas chromatography-mass spectrometry and showed 26 alkyl (C₁ to C₅) carbazoles to be the predominant compounds. An oil-degrading mixed bacterial culture was enriched on carbazole to enhance its ability to degrade nitrogen heterocycles. This culture was used to inoculate a series of flasks of mineral medium and Norman Wells crude oil. Residual oil was recovered from these cultures after incubation at 25°C for various times. The nitrogen-rich fraction was analyzed by capillary gas chromatography, using a nitrogen-specific detector. Most of the C₁-, C₂-, and C₃- carbazoles and one of the C₄-isomers were degraded within 8 days. No further degradation occurred when incubation was extended to 28 days. The general order of susceptibility of the isomers to biodegradation was C₁ > C₂ > C₃ > C₄. The carbazole-enriched culture was still able to degrade n-alkanes, isoprenoids, aromatic hydrocarbons, and sulfur heterocycles in the crude soil.     

Anti-HCV protease of diketopiperazines produced by the Red Sea sponge-associated fungus <Emphasis Type="Italic">Aspergillus versicolor</Emphasis>

Hepatitis C virus (HCV) infection is a global problem due to the difficulties in developing a protective vaccine. In this work, we demonstrated that the ethyl acetate extract of the endophytic fungus Aspergillus versicolor exhibited significant activity against HCV NS3/4A protease with IC₅₀ value of 30 μg/mL. The fungus was isolated from the Red Sea black sponge Spongia officinalis and identified by its morphology and 18S rDNA. Large-scale fermentation of the fungus followed by chromatographic purification with silica gel, Sephadex LH-20 and semipreparative HPLC of the active extract led to isolation of some known metabolites related to cyclodipeptides and the so-called diketopiperazines (DKPs). The DKP, cyclo(L-Tyr-L-Pro), displayed strong effect as HCV protease inhibitor with IC₅₀ value of 8.2 μg/mL. A computational docking study of cyclo(L-Tyr-L-Pro) against HCV protease was used to formulate a hypothetical mechanism for the inhibitory activity of the active compound on the tested enzyme.     

Molecule-size distribution of soluble hiunic compounds from different natural waters

It this study the use of Sephadex G-25, Sephadex LH-20 and CPG-10-75 for the separation of soluble humic compoutids frotn fresh water is tested. It is shown by Sephadex G-25 gel filtration that differences in molecule-size distribution of soluble humic compounds in one lake at different titnes and between lakes can be predicted by E₂₅₀ and E₃₆₅ measurements.     

Metabolism of prostacyclin in cynomolgus monkey

The metabolism of prostacyclin (PGI₂) $\text{in vivo}$ was investigated in Cynomolgus monkey. Following intravenous infusion of 11-[³H]-PGI₂ for three days, pooled urine was extracted with Amberlite XAD-2, then chromatographed and purified by Sephadex LH-20, and reverse phase column chromatography. Radioactive fractions were converted to appropriate derivatives for identification by gas chromatography mass spectrometry. Twelve metabolites were characterized, the major of which was 6-keto-PGF_1α, accounting for 13% of the urinary radioactivity. The metabolic pathways are similar to those observed earlier in the rat. The excretion of substantial amounts of unchanged 6-keto-PGF_1α indicated that the monkey was not able to metabolize PGI₂ as avidly as the rat.     

In Vitro Gibberellin A(1) Binding in Zea mays L

The first and second leaf sheaths of Zea mays L. cv Golden Jubilee were extracted and the extract centrifuged at 100,000g to yield a supernatant or cytosol fraction. Binding of [³H]gibberellin A₁ (GA₁) to a soluble macromolecular component present in the cytosol was demonstrated at 4°C by Sephadex G-200 chromatography. The binding component was of high molecular weight (HMW) and greater than 500 kilodaltons. The HMW component was shown to be a protein and the ³H-activity bound to this protein was largely [³H]GA₁ and not a metabolite. Binding was pH sensitive but only a small percentage (20%) appeared to be exchangeable on addition of unlabeled GA₁. Both biologically active and inactive GAs and non-GAs were able to inhibit GA₁ binding. [³H]GA₁ binding to an intermediate molecular weight (IMW) fraction (40-100 kilodaltons) was also detected, provided cytosol was first desalted using Sephadex G-200 chromatography. Gel filtration studies suggest that the HMW binding component is an aggregate derived from the IMW fraction. The HMW binding fraction can be separated into two components using anion exchange chromatography.     

杠柳枝皮的化学成分研究(英文)

将杠柳枝皮粉末的乙醇提取物悬浮于水后,依次用石油醚、乙酸乙酯和正丁醇萃取.对乙酸乙酯萃取物采用各种柱层析进行多次分离后得到4个化合物,经波谱分析,鉴定它们的结构分别为β-香树素乙酸酯(1)、β-香树素(2)、periplogenin(3)和perilocoside N(4),其中化合物1和2是首次从该植物中分离得到.