Crystallographic and NMR studies on sterols from green alga Chaetomorpha basiretorsa Setchell

Chemical investigation of the ethanol extract of the marine green alga Chaetomorpha basiretorsa Setchell led to the isolation of a new sterol stigmast-4,28-dien-3 alpha 6 beta-diol 1 in addition to the five known sterols of beta-lawsaritol 2, saringosterol 3, 24-hydroperoxy-24-vinyl - cholesterol 4, beta-stigmasterol 5, 29-hydroxystigmasta-5, 24(28) -dien-3 beta-ol 6. Compounds were isolated by normal phase silica gel and Sephadex LH - 20 gel colum chromatography, reverse phase HPLC and recrystalization. Their structures were elucidated by spectroscopic methods including MS, IR 1D/2D NMR and X-ray analysis. Cytotoxicity of compounds was screened by using the standard WIT method. All these compounds were isolated from the green alga Chaetomorpha basiretorsa Setchell for the first time and they were inactive (50% inhibitory concentration was greater than 10 mu g /cm(3)) against KB, Bel -7402, PC - 3M, Ketr 3 and MCF - 7 cell lines.

Steroids from green alga Chaetomorpha basiretorsa Setchell

Six steroids have been isolated from ethanolic extract of green alga Chaetomorpha basiretorsa Setchell by a combination of repeated normal phase silica gel and Sephadex LH-20 gel column chromatography as well as recrystallization. Using spectroscopic methods including MS and NMR, their structures were determined as beta-lawsaritol (1), saringosterol (2), 24-hydroperoxy-24-vinyl-cholesterol (3), beta-stigmasterol (4), stigmast-4-en-3 alpha, 6 beta-diol (5), 29-hydroxystigmasta-5, 24 (28)-dien-3 beta-ol (6). All these compounds were obtained from this genus for the first time and they were inactive (IC50 > 10 mu g /ml) against KB, Bel-7402, PC-3M, Ketr 3 and MCF-7 cell lines.

The chemical constituents from red alga Gymnogongrus flabelliformis Harv.

Eight compounds were isolated from red alga Gymnogongrus flabelliformis Harv. In normal phase silica gel, Sephadex LH-20 gel column chromatography, reverse phase HPLC, and recrystallization. Based on MS and 1D NMR spectroscopic data, their structures were determined as: stigmast-4-en-3-one (I), cholest-4-en-3-one (II), cholesterol (III), uracil (IV), uridine (V), adenosine (VI), succinic acid (VII), and 5-hydroxy-4-methyl-5-pentyl-2,5-dihydro-furan-2-on (VIII). All of them were obtained from this species for the first time. Cytotoxicity of these compounds was screened using standard MTT method, but all the compounds were inactive (IC50 > 10 mu g/ml).

辐状肋柱花的苷类成分

利用硅胶柱层析、Sephadex LH-20及反相硅胶RP-18分离及纯化技术，从辐状肋柱花全草乙醇提取物的正丁醇萃取部分得到7个水溶性成分，经^1H NMR、^13C NMR等波谱技术鉴定为异荭草苷、芒果苷、Swertipunicoside、当药醇苷、异牡荆苷、当药黄素、和7-O-[α-L-吡喃鼠李糖-（1-2）-β-D-吡喃木糖]-1，8-二羟基-3-甲氧基[口山]酮。除异荭草苷外，其余化合物均首次从该植物中得到。

藏药大果大戟中的巨大戟烷型二萜酯类成分

目的研究藏药大果大戟Euphorbia wallichii丙酮提取物中的化学成分。方法用溶剂提取，常规硅胶柱色谱分离和葡聚糖凝胶Sephadex LH-20纯化，采用化学方法和现代波谱分析技术（包括IR，HRESIMS，HRSIMS，1D和2DNMR等）鉴定其化学结构。结果从青海产大果大戟根的丙酮提取物中分离得到6个化合物，分别鉴定为：羊毛甾醇（lanosterol，Ⅰ）、巨大戟二萜-20-肉豆蔻酸酯（ingenol-20-myristinate，Ⅱ）、巨大戟二萜-3-肉豆蔻酸酯（ingenol-3-myristinate，Ⅲ）、没食子酸（gallicacid，Ⅳ）、1-O-a-L-阿拉伯糖-（1→6）-β-D-葡萄糖苷-3，7-二甲基-2-烯-7-羟基-辛醇（1-O-a-L-arabinofuranosyl-（1→6）-β-D-glucopyranosyl-3，7-dimethyl-oct-2-en-7-ol，Ⅴ）、1-O-没食子酰葡萄糖苷（1-O-galloyl-β-D-glucose，Ⅵ）。结论巨大戟烷型二萜酯类化合物Ⅱ和Ⅲ为新化合物，其他化合物均为首次从该植物中分离得到，单萜二糖苷类化合物Ⅴ系首次在该属中发现。

珠芽蓼全草化学成分研究

本文探讨了珠芽蓼全草的化学成分.我们利用硅胶柱多次层析分离和Sephadex LH-20纯化等方法分离得到5个化合物,经NMR,IR,HR-ESI-MS等技术及理化性质鉴定结构,5个化合物分别为β-谷甾醇(β-sitosterol,1)、胡萝卜苷(daucosterol,2)、槲皮素(quercetin,3)、6-O-没食子酰熊果苷(6-O-galloylarbutin,4)、蔗糖(sucrose,5).其中化合物3、4为首次从该植物中分离得到.

珠芽蓼果实化学成分研究

利用硅胶柱层析分离和Sephadex LH20纯化等方法从珠芽蓼果实分离得到6个化合物,经1H NMR、13 C NMR等技术及理化性质鉴定为β-谷甾醇、胡萝卜苷、没食子酸、正丁基-β-D-吡喃果糖苷、槲皮素-5-O-β-D-葡萄糖苷、蔗糖.6种化合物均为首次从该植物分离得到.

尖苞雪莲的化学成分

利用硅胶柱层析、Sephadex LH-20和MCI等分离和纯化手段,从产于青海的菊科风毛菊属植物尖苞雪莲的全草中,分离得到5个化合物,经IR、NMR(1D、2D)、MS等现代波谱技术鉴定它们为芦丁、槲皮素-5-O-β-D-葡萄糖苷、丁香苷、胡萝卜苷和β-谷甾醇,其化学成分为首次报道。

红直獐牙菜的苷类成分

目的：研究红直獐牙菜的苷类成分。方法：采用硅胶柱层析分离和Sephadex LH20纯化，经理化性质和波谱分析鉴定基化学结构。结果：从红直獐牙菜的水溶性部分得到7个化合物，他们分别为8－O－β－D-吡喃葡萄糖－1，5－二羟基－3－甲氧基Shan酮（I）、8－O－β－D－吡喃葡萄糖－1，3，5－三羟基Shan酮（Ⅱ）、1－O－β－D－吡喃葡萄糖－3，7，8－三羟基Shan酮（Ⅲ）、异荭草苷（Ⅳ）、落干酸（V）、龙胆苦苷（Ⅵ）和β－龙胆二糖（Ⅶ）。结论：化合物Ⅲ、Ⅳ、V和Ⅶ为首次从该植物中分得。

The isolation and purification of tris-2,2'-bipyridine complexes of ruthenium(II) containing unsymmetrical ligands

Monomeric ruthenium(II) complexes [Ru(L)3]2+ containing unsymmetric bipyridine ligands [Where L = 5-methyl-2,2'-bipyridine (L1), 5-ethyl-2,2'-bipyridine (L2), 5-propyl-2,2'-bipyridine (L3), 5-(2-methylpropyl)-2,2'-bipyridine (L4), 5-(2,2-dimethylpropyl)-2,2'-bipyridine (L5) and 5-(carbomethoxy)-2,2'-bipyridine (L6)] have been studied and the meridional and facial isomers isolated by the use of cation-exchange column chromatography (SP Sephadex C-25) eluting with either sodium toluene-4-sulfonate or sodium hexanoate. The relative yield of the facial isomer was found to decrease with increasing steric bulk, preventing the isolation of fac-[Ru(L5)3]2+. The two isomeric forms were characterized by 1H NMR, with the complexes [Ru(L1-3)3]2+ demonstrating an unusually large coupling between the H6 and H4 protons. Crystals suitable for X-ray structural analysis of [Ru(L1)3]2+ were obtained as a mixture of the meridional and facial isomers, indicating that separation of this isomeric mixture could not be achieved by fractional crystallisation. The optical isomers of the complex [Ru(L3)3]2+ were chromatographically separated on SP Sephadex C-25 relying upon the inherent chirality of the support. It is apparent that chiral interactions can inhibit geometric isomer separation using this technique.

First report of a novel plant lysozyme with both antifungal and antibacterial activities.

A novel lysozyme exhibiting antifungal activity and with a molecular mass of 14.4 kDa in SDS–polyacrylamide gel electrophoresis was isolated from mung bean (Phaseolus mungo) seeds using a procedure that involved aqueous extraction, ammonium sulfate precipitation, ion exchange chromatography on CM-Sephadex, and high-performance liquid chromatography on POROS HS-20. Its N-terminal sequence was very different from that of hen egg white lysozyme. Its pI was estimated to be above 9.7. The specific activity of the lysozyme was 355 U/mg at pH 5.5 and 30 °C. The lysozyme exhibited a pH optimum at pH 5.5 and a temperature optimum at 55 °C. It is reported herein, for the first time, that a novel plant lysozyme exerted an antifungal action toward Fusarium oxysporum, Fusarium solani, Pythium aphanidermatum, Sclerotium rolfsii, and Botrytis cinerea, in addition to an antibacterial action against Staphylococcus aureus.

Savory peptides present in Moromi obtained from soy sauce fermentation of yellow soybean

The presence of savory peptides in moromi has been investigated. Moromi was prepared by fermenting yellow soybean using Aspergillus oryzae as the starter at the first step (mold fermentation) and 20% brine solution at the next step (brine fermentation). The moromi was then ultrafiltered stepwise using membranes with MW cut-offs of 10,000, 3,000, and 500 Da, respectively. The fraction with MW <500 Da was chromatographed using Sephadex G-25 SF to yield four fractions, 1-4. Analysis of soluble peptides, NaCl content, alpha-amino nitrogen, amino acid composition, peptide profile using CE coupled with DAD, taste profile and free glutamic acid content, were performed for each fraction. Fraction 2 contained a relatively high total glutamic acid content, but a relatively low free glutamic acid content and had the highest umami taste. This fraction also had more peptides containing non-aromatic amino acids than the other fractions. The peptides present in fraction 2 may play a role, at least in part, in its intense umami taste.

Purification of a cathepsin L-like proteinase secreted by adult Fasciola hepatica

A cysteine proteinase released in vitro by Fasciola hepatica was purified to homogeneity by Sephacryl S-200 gel filtration chromatography followed by QAE-Sephadex chromatography. The purified enzyme resolves as a single band with an apparent molecular size of 27 kDa on reducing SDS-polyacrylamide gel electrophoresis; however, under non-reducing conditions it migrates as multiple bands, each with enzymatic activity, in the apparent molecular size range 60-90 kDa. The sequence of the first 20 N-terminal amino acids of the enzyme shows considerable homology with cathepsin L-like proteinases. Immunolocalisation studies revealed that the cathepsin L-like proteinase is concentrated within vesicles in the gut epithelial cells of liver fluke.

Activities of an aqueous plant extract against Herpes simplex virus type 2 (HSV 2): role in virus replication and progeny yield

Tese de mestrado, Biologia Molecular e Genética, Universidade de Lisboa, Faculdade de Ciências, 2015

Carcinoembryonic antigen (CEA) in non digestive cancerous and normal tissues.

Carcinoembryonic antigen (CEA), immunologically identical to CEA derived from colonic carcinoma, was identified and purified from perchloric acid (PCA) extracts of bronchial and mammary carcinoma. CEA extracted from bronchial and mammary carcinoma was quantitated by single radial immunodiffusion and was found to be in average about 50-75 times less abundant in these tumors than in colonic carcinoma. CEA could also be detected in one normal breast in lactation and at lower concentrations in normal lung (1000-4000 times lower than in colonic carcinoma). The small amounts of CEA present in normal tissues are distinct from the glycoprotein of small mol. wt showing only partial identity with CEA, that we recently identified and extracted in much larger quantities from normal lung and spleen. The demonstration of the presence of CEA in non digestive carcinoma by classical gel precipitation analysis suggests that the CEA detected in the plasma of such patients by radioimmunoassay is also identical to colonic carcinoma CEA. Our comparative study of plasma CEA from bronchial and colonic carcinoma, showing that CEA from both types of patient has the same elution pattern on Sephadex G-200 and gives parallel inhibition curves in the radioimmunoassay, is in favor of this hypothesis. However, it should not be concluded that all positive CEA radioimmunoassay indicate the presence of an antigen identical to colonic carcinoma CEA. A word of warning concerning the interpretation of radioimmunoassay is required by the observation that the addition of mg amounts of PCA extract of normal plasma, cleared of CEA by Sephadex filtration, could interfere in the test and mimic the presence of CEA.