中压制备液相色谱快速分离制备儿茶素单体

以反相中压制备液相色谱为工具,甲醇-水作为洗脱溶剂,通过C18 (ODS-AQ)填料从茶多酚中一步分离出表没食子儿茶素(EGC)、表没食子儿茶素没食子酸酯(EGCG)、表儿茶素(EC)、表儿茶素没食子酸酯(ECG)等四种儿茶素单体.1.0g纯度为92.6％的茶多酚经过中压色谱,制备得到了90 mg EGC、355 mg EGCG、23 mgEC和92mg ECG,它们的纯度分别为91.8％、97.6％、97.7％、99.3％,纯品得率56.0％,四种单体总回收率达到68.2％.四种儿茶素单体的结构经核磁共振氢谱、碳谱以及高分辨质谱加以确证.     

杜仲醇的制备及其表征

采用大孔吸附树脂-制备液相色谱联用技术分离制备杜仲醇。以杜仲叶为原料,采用超声波辅助提取,提取液经过正丁醇萃取、D101大孔吸附树脂柱分离纯化后,再通过反相半制备液相色谱分离,以V(甲醇):V(水)=15∶85为流动相进行洗脱,制备得到杜仲醇单体。采用紫外光谱(UV)、红外光谱(IR)、核磁共振波谱(NMR)及液相色谱-质谱联用(LC-MS)等方法对所得单体进行了结构验证。液相色谱法分析检测表明制备所得杜仲醇纯度达95.12%。     

RP-HPLC分离制备桔梗皂苷

在对桔梗总皂苷粗分的基础上,利用制备高效液相色谱分离制备桔梗单体皂苷,共分离得到四个化合物,经波谱法分别鉴定为platycodinD（1）,platycoside E（2）,deapio-platycoside E（3）,未知化合物（4,鉴定中）,经HPLC分析,纯度均大于98％．对比以前的分离手段,该方法分离制备桔梗单体皂苷工艺简单,效率高,为在桔梗中寻找新的天然活性物质提供了新方法。     

应用高速逆流色谱(HSCCC)分离纯化山茱萸中的没食子酸

应用高速逆流色谱（ＨＳＣＣＣ）分离山茱萸中的没食子酸并结合波谱技术进行结构鉴定。经过一次逆流色谱分离,可以得到纯度在７０％以上的没食子酸,第二次分离即可使其纯度达到９７％以上。     

二维制备高效液相色谱分离塞隆骨强极性多肽

本实验建立了一种新型的利用高正交性的二维制备型高效液相色谱系统分离强极性动物药多肽的方法。本文以塞隆骨水提取物为研究对象,以亲水性C18AQ制备型高效色谱柱为第一维分离柱,首先在一维分离中将目标混合物分成若干组份;然后以C18MP制备型高效液相色谱柱为第二维色谱分离柱,将第一维分离后得到的组份纯化为单体化合物。本研究最终得到5个塞隆骨单体化合物,化合物纯度均超过98%。经Nano-LCESI-MS/MS鉴定和搜库分析,这些多肽的序列分别为:KTAILVKE、RGAPQDQE、LVGPGAPGR、GFAGD和KPQWHP。此研究方法速度快、效率高且重复性好,可以对类似的研究提供借鉴。     

二维制备高效液相色谱分离塞隆骨强极性多肽北大核心CSCD

本实验建立了一种新型的利用高正交性的二维制备型高效液相色谱系统分离强极性动物药多肽的方法。本文以塞隆骨水提取物为研究对象,以亲水性C18AQ制备型高效色谱柱为第一维分离柱,首先在一维分离中将目标混合物分成若干组份;然后以C18MP制备型高效液相色谱柱为第二维色谱分离柱,将第一维分离后得到的组份纯化为单体化合物。本研究最终得到5个塞隆骨单体化合物,化合物纯度均超过98%。经Nano-LCESI-MS/MS鉴定和搜库分析,这些多肽的序列分别为:KTAILVKE、RGAPQDQE、LVGPGAPGR、GFAGD和KPQWHP。此研究方法速度快、效率高且重复性好,可以对类似的研究提供借鉴。     

蝙蝠葛的化学成分研究

目的:对防己科蝙蝠葛属植物蝙蝠葛(Menispermum dauricum DC)的干燥根茎化学成分进行研究.方法:采用抗肿瘤细胞活性实验与中药化学实验相结合的方法,最终确定蝙蝠葛95％乙醇提取物,再经大孔树脂以70％乙醇溶液洗脱部位为其抗肿瘤活性部位,再经硅胶柱色谱法、Sephadex LH-20凝胶柱色谱法、中低压液相色谱及高效液相色谱法等现代分离技术对其化学成分进行系统研究,分离得到单体化学成分,并通过其理化性质与波谱数据(Uv、IR、NMR、MS等)分析鉴定其化学结构.结果:从蝙蝠葛药材95％乙醇提取物中分离得到7个化学成分,分别鉴定为蝙蝠葛新诺林碱(1)、northalifoline(2)、青藤碱(3)、粉防己碱(4)、荷苞牡丹碱(5)、紫堇定(6)、香草酸(7).结论:其中化合物5、6、7均为首次从该属植物中分离得到.     

从马钱子中选择分离马钱子碱新方法的研究

本文采用分子模板聚合物技术对中草药马钱子(Strychnos nux-uomica L.)粗提物中有效成分马钱子碱(brucine)、士的宁(strychnine)进行了选择性分离。用丙烯酸为单体,TMPTA为交联剂,合成对马钱子碱有特异选择性的模板聚合物(MIP)。选择性分离出粗提物中的马钱子碱,纯度可达99．7％,同时得到士的宁为主的生物碱,纯度为94．6％。     

极地海洋真菌赛氏曲霉活性成分研究

通过对菌株r DNA-ITS序列和系统发育多样性进行分析,鉴定该菌株为赛氏曲霉;对赛氏曲霉用大米培养基进行固体发酵培养,对其发酵产物采用有机溶剂萃取、硅胶柱色谱层析、反相ODS柱色谱层析、薄层色谱层析、Sephadex LH-20凝胶柱色谱层析和制备型高效液相色谱等分离纯化手段进行分离,得到7个单体化合物。对分离得到单体化合物采用质谱、核磁共振氢谱、核磁共振碳谱、二维核磁共振等现代波谱技术对其进行结构鉴定;对单体化合物的MIC值进行测定,其中化合物L-1对耐甲氧西林金黄色葡萄球(MRSA)、耐甲氧西林表皮葡萄球菌(MRSE)有一定的抑菌活性,对大肠杆菌有明显的抑菌活性。     

凝胶色谱法制备寡糖

采用已知结构的多糖,控制其水解条件,使之产生所需寡糖片断。用聚丙烯酰胺凝胶色谱(BiO-6el P-4)分离,对中性糖可以分离到含十一个糖残基的寡糖。对氨基糖可分离到含七个糖残基的寡糖。用薄层色谱和快原子轰击质谱鉴定了它们的纯度。     

Comparison of the antioxidant action of the alcoholic extract of Rubia cordifolia with rubiadin.

The inhibition of FeSO4 induced lipid peroxidation in rat liver by alcoholic extract of Rubia cordifolia and by one of its constituent rubiadin (1, 3-dihydroxy-2-methyl anthraquinone) (pure form) has been compared. Both have been found to inhibit lipid peroxidation in a dose dependent manner. Whereas the former shows both oxidising and reducing properties with Fe2+ and Fe3+, the latter shows oxidising property only by converting Fe2+ to Fe3+. The former inhibits the oxidation of reduced glutathione while the latter does not.     

Flavonoids from Wyethia glabra

Ten flavonoid compounds, including three new natural products, were isolated from a dichloromethane extract of Wyethia glabra. The known compounds are: orobol 7-methyl ether, orobol 3′-methyl ether, naringenin 7-methyl ether, eriodictyol, 8-C-prenyleriodictyol, 6-C-prenyleriodictyol and 8-C-prenylnaringenin. Eriodictyol 7-methyl ether, 2′,4′,6′-trihydroxy-4-methoxychalcone and 6-C-prenylnaringenin are new natural products. An additional prenylated flavanone was isolated and partially characterized.     

Anthraquinones from Hedyotis capitellata

Four new furanoanthraquinones, 2-hydroxymethyl-3,4-[2'-(1-hydroxy-1-methylethyl)-dihydrofurano]-8-hydroxyanthraquinone, 2-hydroxymethyl-3,4-[1'-hydroxy-2'-(1-hydroxy-1-methylethyl)-dihydrofurano]-8-hydroxyanthraquinone, 2-hydroxymethyl-3,4-[2'-1-hydroxy-1-methylethyl)-dihydrofurano]anthraquinone and 2-methyl-3,4-[2'-(1-hydroxy-1-methylethyl)-dihydrofurano] anthraquinone or capitellataquinone A-D and four known anthraquinones, rubiadin, anthragallol 2-methyl ether, alizarin 1-methyl ether and digiferruginol, together with scopoletin were isolated from the stems of Hedyotis capitellata Wall (Rubiaceae). Lucidin-3-O-beta-glucoside was isolated from the roots of the plant. Characterization of the new compounds was carried out by extensive NMR studies using FGCOSY, FGHMQC, FGHMBC and DEPT-135 in addition to other spectroscopic methods.     

3-Hydroxyisoflavanones from the stem bark of Dalbergia melanoxylon: Isolation,antimycobacterial evaluation and molecular docking studies

Two new 3-hydroxyisoflavanones, (S)-3,4′,5-trihydroxy-2′,7-dimethoxy-3′-prenylisoflavanone (trivial name kenusanone F 7-methyl ether) and (S)-3,5-dihydroxy-2′,7-dimethoxy-2″,2″-dimethylpyrano[5″,6″:3′,4′]isoflavanone (trivial name sophoronol-7-methyl ether) along with two known compounds (dalbergin and formononetin) were isolated from the stem bark of Dalbergia melanoxylon. The structures were elucidated using spectroscopic techniques. Kenusanone F 7-methyl ether showed activity against Mycobacterium tuberculosis, whereas both of the new compounds were inactive against the malaria parasite Plasmodium falciparum at 10 μg/ml. Docking studies showed that the new compounds kenusanone F 7-methyl ether and sophoronol-7-methyl ether have high affinity for the M. tuberculosis drug target INHA.     

Hydropiperoside,a novel coumaryl glycoside from the root of Polygonum hydropiper

From the methanol extract of the root of Polygonum hydropiper, a novel coumaryl glycoside hydropiperoside was isolated together with anthraquinone, ellagic acid 3,3′-di-O-methyl ether, gallic acid, two quercetin glycosides and an unidentified aromatic δ-lactone possessing antifertility activity. The structure of hydropiperoside was established as β-d-(1,3,6-tri-p-coumaryl)-fructofuranosyl-α-d-glucopyranoside by combination of extensive ¹H NMR and ¹³C NMR spectra, and the FD/MS spectrum.     

红厚壳枝条的化学成分

从红厚壳(Calophyllum inophyllum Linn.)枝条乙醇提取物中分离得到11个化合物,通过光谱分析,鉴定其结构为:6-羟基-2,3-二甲氧基 酮 ( 1 ) ,1,3,7-三羟基 酮 ( 2 ) ,1,3,7-三羟基-8-甲氧基 酮( 3 ) ,7-羟基-1,3-二甲氧基 酮( 4 ) ,伪蒲公英甾醇( 5 ) ,1,3,6-三羟基-5,7-二甲氧基 酮( 6 ) ,2-羟基-1-甲氧基 酮 ( 7 ) ,2-羟基-1,8-二甲氧基 酮 ( 8 ) ,1,3,5-三羟基-2-甲氧基 酮 ( 9 ) ,4-羟基 酮 ( 10 ) ,1,3,5-三羟基 酮 ( 11 ) 。化合物 2 ~ 5 为首次从红厚壳属植物中得到,化合物 6 ~ 8 为首次从该植物中得到,化合物 1 为一新的天然产物。细胞毒活性测试结果表明,化合物 9 对人胃癌细胞(SGC-7901)的增殖显示出生长抑制活性, 其IC₅₀值为1.8×10^-5 mol/L。     

Flavonoids from Artemisia ludoviciana var. ludoviciana

Nineteen flavonoids were isolated from Artemisia ludoviciana var. ludoviciana, including a new 2′- hydroxy- 6-methoxyflavone, 5,7,2′,4′-tetrahydroxy-6,5′-dimethoxyflavone. The known compounds include quercetagetin 3,6,3′,4′-tetramethyl ether, eupatilin, 5,7-dihydroxy-3,6,8,4′-tetramethoxyflavone, luteolin 3′,4′-dimethyl ether, jaceosidin, 5,7,4′-trihydroxy-3,6-dimethoxyflavone, tricin, hispidulin, chrysoeriol, kaempferol 3-methyl ether, apigenin, axillarin, eupafolin, selagin and luteolin together with three flavones which were previously isolated for the first time from Artemisia frigida: 5,7,4′-trihydroxy-6, 3′,5′-trimethoxyflavone, 5,7,3′-trihydroxy-6,4′,5′-trimethoxyflavone and 5,7,3′,4′-tetrahydroxy-6,5′- dimethoxyflavone.     

Induction of anthraquinone biosynthesis in Rubia cordifolia cells by heterologous expression of a calcium-dependent protein kinase gene

Calcium-dependent protein kinases (CDPKs) play an important role in plant cell responses to stress and pathogenic attack. In this study, we investigated the effect of heterologous expression of the Arabidopsis CDPK gene, AtCPK1, on anthraquinone production in transgenic Rubia cordifolia cells. AtCPK1 variants (a constitutively active, Ca(2+) -independent form and a non-active form used as a negative control) were transferred to callus cells by agrobacterial transformation. Overexpression of the constitutively active, Ca(2+) -independent form in R. cordifolia cells caused a 10-fold increase in anthraquinone content compared with non-transformed control cells, while the non-active form of AtCPK1 had no effect on anthraquinone production. Real-time PCR measurements showed that the activation of anthraquinone biosynthesis in transgenic calli correlated with the activation of isochorismate synthase gene expression. The activator effect of AtCPK1 was stable during prolonged periods of transgenic cell cultivation (more than 3 years) and the transgenic cultures exhibited high growth. Our results provide the first evidence that a CDPK gene can be used for the engineering of secondary metabolism in plant cells.     

Pharmacological and toxicological activity of Heterophyllaea pustulata anthraquinone extracts

Benzenic extracts from both stems and leaves of Heterophyllaea pustulata showed the most significant activity in vivo in the Brine Shrimp Lethally Test (BST), relative to others of different polarity. They were therefore selected for in vitro antimicrobial activity studies. Bacteriostatic activity against Micrococcus luteus ATCC 9341 was detected, selectively inhibiting both oxacillin-sensitive and -resistant Staphylococcus aureus, among several gram-positive and gram-negative bacterial species tested. Antifungal activity against important opportunist microorganisms and against those involved in superficial mycosis, all from nosocomial origin was also detected. A chemical screening revealed the presence of anthraquinones as major compounds. Among them, we identified damnacanthal, rubiadin, 2-hydroxy-3-methyl anthraquinone, soranjidiol, rubiadin-1-methyl ether, and damnacanthol in the benzenic stem extract. The benzenic leaf extract shows a similar chemical composition, except for damnacanthal, damnacanthol, soranjidiol-1-methyl ether, and 3 anthraquinones whose structures have not yet been elucidated. Acute toxicity studies revealed a low toxicity in mice for the anthraquinonic extracts, as measured in the LD50 value (123 mg/kg body wt. i.v.), and death was not observed at doses of up to 4000 mg/kg body wt. s.c.     

茜草叶绿体全基因组序列及其系统发育分析

以茜草科(Rubiaceae)植物茜草(Rubia cordifolia L.)为研究对象，采用Illumination高通量测序技术对其叶绿体全基因组进行测序，并运用MAGA11等生信学工具进行全基因组解析及系统发育分析。结果表明，(1)茜草叶绿体基因组呈典型的四分环状结构，总GC含量37.2%,长153 959 bp,其中包含大单拷贝(LSC)区83 844 bp、小单拷贝(SSC)区17 083 bp和2个反向重复(IR)区26 516 bp;共注释得到124个基因，包括79个蛋白质编码基因、37个tRNA和8个rRNA。(2)序列共鉴定到169个SSR位点，以A、T组成为主，包括129个单核苷酸、18个双核苷酸、11个三核苷酸、9个四核苷酸和2个五核苷酸，六核苷酸SSR未检测到；边界分析显示，茜草叶绿体基因组LSC区差异性最大，变异程度最高，而IRa区则差异性最小，最为保守。(3)最大似然法(ML)构建系统发育树显示，样品茜草与同属植物Rubia horrida以100%支持率聚为一类，茜草亚科、仙丹花亚科与金鸡纳亚科聚为姐妹类群，证明茜草科植物在进化过程中保守发育。