黄疣海参皂苷 hillaside A 和 hillaside B 的体外抗真菌及抗肿瘤活性

目的阐明黄疣海参hillasideA和hillasideB的抗肿瘤及体外抗真菌活性。方法对黄疣海参的总提取物采取硅胶柱色谱及半制备高效液相色谱（HPLC）法等分离手段进行活性成分追踪分离,通过理化性质及波谱学手段进行化学结构鉴定,以磺酰罗丹明B蛋白染色（SRB）法评价化合物的抗肿瘤活性。结果分离得到了2个新的三萜皂苷元类化合物,命名为hillasideA和hillasideB。2个新化合物均显示了较强的体外抗真菌及抗肿瘤活性。结论研究为研制新的抗肿瘤药物提供了先导化合物,为充分开发利用我国的海洋生物资源提供了一定的物质基础和科学依据。     

深海来源曲霉属真菌CXCTD-06-6a次级代谢产物中的肿瘤细胞增殖抑制活性成分研究

目的 对一株来源于深海的曲霉属真菌CXCTD-06-6a发酵产物中的抗肿瘤活性成分进行分离和鉴定.方法 采用溶剂萃取、柱色谱层析及制备HPLC等方法对菌株发酵产物的活性部位进行了活性追踪分离,通过理化性质及波谱学手段进行化学结构鉴定,以MTT法评价了化合物的抗肿瘤活性.结果 从菌株CXCTD-06-6a的发酵产物中分离...     

夏枯草植物内生真菌CPCC 480171抗肿瘤活性成分的研究

利用硅胶柱色谱、凝胶色谱、制备高效液相色谱等方法,对一株分离自夏枯草内生真菌中具抗肿瘤活性代谢产物进行研究,从菌株发酵产物中分离得到单组份,根据理化性质及有机波谱鉴定其化学结构.结果:分离得到3个活性化合物,结构鉴定分别为:链格孢毒素Altertoxin-I,4',5,7-三羟基异黄酮,4',7-二羟基异黄酮.对其进行了抗肿瘤活性实验.分离得到的3个化合物均具有不同程度的抗肿瘤作用.     

唐松草属生物碱ⅩⅩ.长柱唐松草根中生物碱的分离、成份与结构鉴定

唐松草属植物富有生物碱并具有降压、抗菌及抗肿瘤活性,已研究了36种不同的唐松草属植物,并至少分离到125种生物碱。长柱唐松草(Thalictrum longistylum     

灵芝多糖分离鉴定及抗肿瘤活性的研究

目的探讨灵芝多糖的分离纯化、理化性质及初步抗肿瘤活性。方法采用热水提取 ,乙醇分级沉淀 ,DEAE 32离子交换柱色谱等方法从灵芝子实体中分离得到灵芝多糖。用SephadexG 10 0凝胶柱色谱法测定其纯度和平均分子量。用薄层色谱及气相色谱法测定其单糖组成。溴化四唑蓝法测定其初步体外抗肿瘤活性。结果从灵芝子实体中分离得到 4种均一多糖 (GLPL1 ～GLPL4 ) ,其中GLPL1 和GLPL3为主要成分 ,其分子量为 4 10 0和 5 70 0。GLPL1 由葡萄糖组成 ;GLPL3由葡萄糖和半乳糖组成。GLPL1 和GLPL3对人鼻咽癌细胞增殖有显著的抑制作用 ,GLPL3还对人胃癌细胞、人结肠癌细胞增殖有一定的抑制作用。结论灵芝多糖GLPL1 、GLPL3是很有潜力的抗肿瘤或辅助抗肿瘤药物     

茯苓抗肿瘤成分研究(Ⅰ)

用硅胶柱层析法从茯苓Poria cocos(Schw.)Wolf的干燥菌核中分离出一种三萜类有机酸,根据理化性质及光谱数据确定为茯苓酸(Pachymic acid),抗肿瘤实验表明茯苓酸对小鼠Sarcoma 180抑制率达62.8％,茯苓中的水溶性低分子量化合物无抗肿瘤活性。     

海洋来源的放线菌3295代谢产物的结构鉴定及抗肿瘤活性(Ⅰ)

目的 对海洋来源的放线菌3295代谢产物中的抗肿瘤活性成分进行分离和鉴定。方法 利用细胞周期抑制为抗肿瘤活性指标，采用Sephadex LH—20．硅胶和HPLC等柱色谱技术对3295活性菌株发酵物进行活性追踪分离，根据理化性质和光谱方法进行化学结构鉴定，用流式细胞术评价化合物的抗肿瘤活性。结果与结论 从其代谢产物中分离得到1个具有抗肿瘤活性的化合物I，结构鉴定为邻苯二甲酸二丁酯(dibutylphthalate)；化合物Ⅰ对小鼠乳腺癌温敏型tsFT210细胞具有G0／G1期细胞周期抑制作用。     

泽漆的化学成分及其抗肿瘤转移活性研究

目的 研究泽漆石油醚、醋酸乙酯部位的化学成分及其抗肿瘤转移活性。方法 泽漆全草采用50%丙酮提取、系统溶剂萃取,综合运用硅胶柱色谱、凝胶柱色谱、HPLC制备色谱等方法分离纯化泽漆石油醚、醋酸乙酯部位的化学成分;采用根据理化性质和波谱分析方法鉴定其结构;通过MTT实验筛选出非细胞毒性的化学成分,进一步通过划痕实验来评价酚酸类化合物抗肿瘤转移的活性。结果 从泽漆石油醚、醋酸乙酯部位分离鉴定了8个化合物,分别鉴定为没食子酸（1）、咖啡酸（2）、间苯二酚（3）、连苯三酚（4）、没食子酸甲酯（5）、没食子酸乙酯（6）、杨梅素（7）、槲皮素（8）。抗肿瘤转移活性筛选结果表明,化合物1、5、6均能剂量相关性地提高抗肿瘤转移活性,其IC50分别为0.475、4.568、4.612 μmol/L。结论 化合物1、5、6显示出较好的抗肿瘤细胞转移活性,其中化合物1的作用最为显著。     

抗癌散的抗肿瘤活性成分研究

目的：对抗癌散的抗肿瘤活性成分进行研究。方法：用硅胶、葡聚糖凝胶（Sephadex LH-20）柱色谱等方法进行分离纯化,通过理化性质及光谱数据确定化合物的结构;采用流式细胞术（FCM）对部分单体化合物进行抗肿瘤活性的测定。结果：分离得到β-谷甾醇（Ⅰ）、正丁基-β-D-吡喃果糖苷（Ⅱ）、人参皂苷Rg1（Ⅲ）3个化合物;活性实验结果表明,部分化合物具有一定的抗肿瘤活性。结论：本研究初步阐明了复方抗癌散中的主要化学成分,其中化合物Ⅱ为首次从该植物中分离得到;化合物（Ⅱ）和化合物（Ⅲ）对人肝癌细胞（Bel-7402）有较好的抑制活性。     

海洋放线菌分离及菌株Streptomyces sp. AH17-3的次级代谢产物研究

目的对海洋放线菌进行分离及抗肿瘤活性筛选,并对一株具有抗肿瘤活性的海洋放线菌AH17-3的次级代谢产物进行研究。方法采用溶剂萃取、柱色谱层析及制备HPLC等方法对菌株AH17-3的发酵产物进行化学分离,通过理化性质及波谱学方法并参阅文献进行化合物结构鉴定,以SRB法评价化合物的抗肿瘤活性。结果从海洋样品中分离放线菌174株,从菌株AH17-3中分离得到了4个聚酮类化合物,经鉴定其结构分别为germicidin A(1)、germicidin B(2)、daidzein(3)、genistein(4)。其中化合物1具有弱的细胞毒活性,其IC50为3.5×10-7 M。结论海洋放线菌是重要的药用微生物资源,化合物1,2均为首次从海洋放线菌中分离得到。     

应用国产色谱柱测定胺碘酮血浆浓度的研究

本文建立了在进口高效液相色谱仪上应用国产色谱柱取代进口色谱柱测定胺碘酮血浓度的方法。两法的最低检测限均为20ng,血浆中最低有效检测浓度均为0.2μg/ml,日内回收率国产色谱柱法为95.8%,进口色谱柱法为96.5%.日间回收率前法为95.0%,后法为96.7%.两法测定同一血药浓度接近,相关系数为0.999,斜率为0.9852,结果表明国产色谱柱可代替进口色谱柱用于血浆胺碘酮浓度监测。     

Determination of terbutaline enantiomers in human plasma by coupled achiral-chiral high performance liquid chromatography

Achiral-chiral column switching HPLC assay was developed to allow the separation and quantification of the enantiomers of terbutaline in human plasma by means of fluorescence detection. Plasma samples were prepared by solid-phase extraction with sep-pak silica, followed by HPLC assay. The enantiomers of terbutaline and the internal standard were separated from the biological matrix on a silica column, and the two enantiomers were resolved and quantified on a Sumichiral OA-4900 column. The two columns were connected by a switching valve equipped with silica trap column. The trap column was used to concentrate the terbutaline in the eluent from the achiral column before back flushing onto the chiral phase. For each enantiomers, the assay was linear between 2.5-125 ng/ml (r=0.9999) and detection limit was 1.0 ng/ml.     

Fractional determination of bile acids bound with protein in serum by high-performance liquid chromatography using dual-column switching system

A simple and rapid technique for the fractional determination of bile acids bound with protein in the serum was developed by using high performance liquid chromatography with a dual-column switching system. The serum samples were directly injected onto a first column (hydroxyapatite), which was initially flushed with 1 mM phosphate buffer. Serum proteins were strongly retained on a column of hydroxyapatite, but free bile acids were not retained. The bile acids were adsorbed on a second column (Serumout-25) and eluted onto a column of immobilized 3 alpha-hydroxysteroid dehydrogenase (3 alpha-HSD) with an elution solvent (CH3CN-MeOH-30 mM ammonium acetate, 30:30:40, v/v/v). Reduced nicotinamideadenine dinucleotide was produced on the immobilized 3 alpha-HSD column and then determined fluorometrically. Subsequently, the hydroxyapatite column was flushed with 20 mM phosphate buffer. Bile acids bound with albumin were eluted and condensed on Serumout-25. The phosphate buffer (400 mM) was finally used for the elution of bile acids bound with globulin from the hydroxyapatite column. Each condensed bile acid was eluted onto the immobilized 3 alpha-HSD column as described above.     

Determination of microcystins in lake water using reusable immunoaffinity column.

A reusable immunoaffinity column for purification of microcystins in lake water was prepared by coupling anti-microcystin-LR monoclonal antibodies to immunoaffinity support. Thanks to spherical shape of the immunoaffinity support Formyl-Cellulofine used in this study, applied solutions passed the column smoothly even when used repeatedly. Reusability of the column was examined by determining the recoveries of spiked microcystins-RR, -YR and -LR (100ng each) from lake water. After extraction with a Sep-Pak PS2 cartridge containing styrene-divinylbenzene copolymer, the extract was purified with the immunoaffinity column. The immunoaffinity column was regenerated by washing with Tris-HCl buffer containing bovine serum albumin for repeated uses. Recoveries of spiked microcystins from the first use of the column were 87-88%, and 83-88% from the second and third uses, and the recoveries gradually dropped to 63-77% from the 4-5th uses, the results of which indicated that the column could be used repeatedly for three times. The present method was applied to determine microcystins in water collected from three different lakes in Japan in 1999. In a sample from Lake Suwa, microcystins-RR and -LR were determined by high performance liquid chromatography with photodiode array detection and electrospray ionization-liquid chromatography/mass spectrometry.     

Pharmacokinetic study of beta-methyldigoxin by enzyme immunoassay using a novel specific antiserum in rats

We previously showed that enzyme immunoassay (EIA) of beta-methyldigoxin (MDx3) using anti-MDx3 3'-hemisuccinate-bovine serum albumin antiserum (Antiserum-I) was superior to that using commercial anti-digoxin antiserum (Antiserum-II) in terms of specificity and that pretreatment of human serum with phenyl boric acid (PBA) column was effective. In the present study, we examined the precision of EIA using Antiserum-I and the recovery of MDx3 after PBA column treatment in rat serum, and also investigated pharmacokinetic changes of MDx3 in rats. The intra- and inter-assay variations and recovery tests using Antiserum-I were good. The PBA column was effective in selectively separating MDx3 from rat serum containing MDx3 and its metabolites. The recovery tests using Antiserum-I with PBA column showed about 110% and the interference of metabolites of MDx3 was negligible. Serum concentration-time courses of MDx3 by EIA using Antiserum-I with PBA column and Antiserum-I were lower than that using Antiserum-II. The distribution volume at steady state and total body clearance values of MDx3 in these conditions were significantly higher than those using Antiserum-II. The usefulness of PBA column was ascertained, while effects of PBA column on these parameters were not significant. In addition, rapid absorption of MDx3 was observed by EIA using Antiserum-I with PBA column. These results suggest that EIA using Antiserum-I with PBA column for the pretreatment of serum samples should be a more useful and valuable system in therapeutic drug monitoring and pharmacokinetic studies of the unchanged type of MDx3 than Antiserum-II.     

Potentials of wide-bore fused silica capillary columns for substance identification by means of retention indices

A wide-bore capillary column with a methylsilicone stationary phase has been evaluated with regard to its ability to identify substances by means of their retention indices. Column efficiency was compared with a normal packed column and a medium-bore capillary column, and the impact of carrier gas flow and load capacity were investigated. It was shown that under defined conditions retention indices on the wide-bore capillary column were comparable to those available in a data bank obtained on packed columns.     

两种气相色谱法测定珍珠明目滴眼液中冰片的含量

目的 比较两种GC法测定珍珠明目滴眼液中冰片含量的差异。方法同一样品采用填充柱与毛细管柱分别测定。结果两种方法的精密度和准确度均较好,测定结果接近。结论考虑到分析周期及分离效果,建议用毛细管柱取代传统的填充柱测定样品中冰片的含量。     

Liquid chromatographic resolution of the isomers of tipredane and phenylthioproline using urea-solubilized beta-cyclodextrin in the mobile phase.

Two LC assays were developed using urea-solubilized beta-cyclodextrin (beta-CD) as a mobile phase additive in combination with reversed-phase columns. A methylsilane column gave optimal resolution of the steroid tipredane from its epimer. An investigation of the effects of beta-CD concentration, column temperature, column type, and addition of ethanol on the chromatographic separation is detailed. The enantiomer and diastereoisomers of L-cis-phenylthioproline were best resolved by urea-solubilized beta-cyclodextrin and a trimethylsilane column. The elution order of L-cis-phenylthioproline relative to its stereoisomers was reversed after adding ethanol to the beta-CD containing mobile phase or by changing from a beta-CD to an acetylated beta-CD column. The resolution factors for these separations obtained using the beta-CD mobile phase were larger than those obtained using beta-CD columns. Mobile phases containing up to 0.15 M beta-CD and 8 M urea were investigated. The separation of these isomers are dramatically affected by column polarity.     

A mathematical model to predict the size of the pellets formed in freeze pelletization techniques: parameters affecting pellet size

A mathematical model was developed based on the theory of drop formation to predict the size of the pellets formed in the freeze pelletization process. Further the model was validated by studying the effect of various parameters on the pellet size such as viscosity of the pellet forming and column liquids, surface/interfacial tension, density difference between pellet forming and column liquids; size, shape, and material of construction of the needle tips and temperatures maintained in the columns. In this study, pellets were prepared from different matrices including polyethylene glycols and waxes. The column liquids studied were silicone oils and aqueous glycerol solutions. The surface/interfacial tension, density difference between pellet forming and column liquids and needle tip size were found to be the most important factors affecting pellet size. The viscosity of the column liquid was not found to significantly affect the size of the pellets. The size of the pellets was also not affected by the pellet forming liquids of low viscosities. An increase in the initial column temperature slightly decreased the pellet size. The mathematical model developed was found to successfully predict the size of the pellets with an average error of 3.32% for different matrices that were studied.     

GC测定细辛药材中的农药残留量

目的:建立细辛药材中10种农药残留的气相色谱分析方法。方法:样品经丙酮-水(7∶3,v/v)超声提取后,用正己烷进行液液分配,提取液经弗罗里硅土柱净化,采用OV-1701弹性石英毛细管柱进行分离,用毛细管气相色谱-电子捕获检测器(GC-ECD)同时检测。结果:各农药回归方程的线性良好,相关系数为0.9903～0.9991;仪器精密度为1.2%～3.4%;方法精密度为6.5%～7.4%;加样回收率为84.0%～103.2%,RSD为2.5%～8.8%。结论:本方法可用于中药材中农药残留的测定。