高速逆流色谱分离小孢拟盘多毛孢PM-1 菌株中的除草活性物质

【目的】应用高速逆流色谱法(High-speed counter-current chromatography,HSCCC)实现对小孢拟盘多毛孢Pestalotiopsis microspora PM-1菌株代谢产物中除草活性物质的首次分离。【方法】以正己烷:乙酸乙酯:甲醇:水(4:5:4:5,V/V/V/V)为最佳的两相溶剂体系,上相(水相)为固定相,下相(有机相)为流动相,正相洗脱。【结果】在流速2 mL/min、转速900 r/min、检测波长254 nm的条件下进行分离,得到4个馏分,其中馏分Ⅱ对马唐的活性较强。馏分Ⅱ经高效液相色谱法(High performance liquid chromatography,HPLC)以乙腈:水=75:25(V/V)为流动相经过C18柱进一步分离检测,所得馏分A对马唐种子萌发有较强的抑制作用。其保留时间为7.954 min,该峰经二极管阵列光谱检测为单一组分。【结论】利用最佳的高速逆流色谱条件和高效液相色谱条件对小孢拟盘多毛孢Pestalotiopsismicrospora PM-1菌株代谢产物进行分离,可获得除草活性化合物——馏分A。     

黄皮种子提取物对斜纹夜蛾幼虫的杀虫活性及有效成分鉴定

【目的】测定黄皮Clausena lansium种子提取物对斜纹夜蛾Spodoptera litura幼虫的拒食活性,对有效成分进行分离、鉴定,并进一步测定其拒食活性、生长发育抑制活性及细胞毒性,为筛选斜纹夜蛾生态防治植物源杀虫剂提供依据。【方法】采用叶蝶法测定黄皮种子甲醇提取物及其石油醚、乙酸乙酯、正丁醇和水4种溶剂萃取物对斜纹夜蛾3龄幼虫的非选择性拒食活性和生长发育抑制活性;用硅胶柱层析进行石油醚萃取物和乙酸乙酯萃取物拒食活性成分分离;用核磁共振谱(NMR)进行拒食有效成分结构鉴定;用MTT法测定拒食活性成分对SL细胞毒性。【结果】黄皮种子甲醇提取物对斜纹夜蛾3龄幼虫具有较强的拒食活性,活性成分主要分布于石油醚相和乙酸乙酯相中;石油醚相和乙酸乙酯相经硅胶柱层析分离得10个馏分,以黄色晶体馏分2的拒食活性最强;经分离纯化及核磁共振氢谱和碳谱鉴定,黄皮新肉桂酰胺B(lansiumamide B)为主要的拒食活性成分,对斜纹夜蛾3龄幼虫48 h拒食中浓度为214.95μg/m L,同时具有较强的细胞毒性,对SL细胞48 h抑制中浓度为10.09μg/m L。【结论】黄皮种子提取物对斜纹夜蛾幼虫具有较强的拒食活性及细胞毒性,黄皮新肉桂酰胺B是主要活性成分之一,作为植物源杀虫剂开发具有较好的发展前景。     

黄连提取物对α-葡萄糖苷酶抑制作用研究

利用体外α-葡萄糖苷酶抑制模型对黄连不同部位提取物进行活性评价,并与阳性对照Acarbose比较,发现黄连不同部位均有一定的α-葡萄糖苷酶抑制活性.其中,黄连根茎乙酸乙酯提取物的抑制活性最高(IC_(50)=20.72 μg/mL),黄连种子石油醚部位(IC_(50)=40.86 μg/mL)和黄连叶石油醚部位(IC_(50)=62.85 μg/mL)的活性次之.3个部位的提取物活性均远大于阳性对照Acarbose(IC_(50)=1081,27 μg/mL).不同部位比较,根茎对α-葡萄糖苷酶抑制活性最好,这3种提取物抑制活性均比阳性对照高;同一部位不同提取物比较,石油醚和甲醇提取物α-葡萄糖苷酶抑制活性一般要高于乙酸乙酯提取物.     

凹叶厚朴抑制α-糖苷酶与抗氧化活性研究

首次采用96微孔板法检测贵州和河南产凹叶厚朴抑制α-葡萄糖苷酶活性;并采用DPPH、ABTS和FRAP三种方法测定其抗氧化活性.贵州产凹叶厚朴乙酸乙酯(IC50 =7.22 μg,/mL)和正丁醇提取部位(IC50=36.59 μg/mL),河南产凹叶厚朴石油醚(IC50=107.04 μg/mL)和乙酸乙酯提取部位(IC50=17.17μg/mL),它们的活性都远高于于阳性对照Acarhose( IC50=1081.27 μg/mL).贵州产凹叶厚朴乙酸乙酯提取部位清除ABTS自由基的能力最强(IC50=8.81 μg/mL),强于阳性对照BHT(IC50=11.94 μg/mL);其次为河南产凹叶厚朴乙酸乙酯提取部位(IC50=12.73 μg/mL).研究结果表明,贵州产凹叶厚朴乙酸乙酯提取部位抑制α-葡萄糖苷酶和抗氧化活性最好.     

八角茴香提取物对长角扁谷盗成虫的熏蒸作用及其酶活性的影响

为探索八角茴香的杀虫活性及其作用机理,采用密闭容器熏蒸法测定八角茴香干果的甲醇、乙酸乙酯、石油醚3种溶剂提取物对长角扁谷盗成虫的杀虫活性,及其对试虫体内乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GSTs)活性的影响。结果表明:3种提取物对试虫均有较强的熏杀活性,熏杀效果随着处理剂量的提高和处理时间的延长而增强,熏蒸72 h后,20.0 mg/L甲醇、乙酸乙酯、石油醚提取物对试虫的校正死亡率分别为89.93%、96.71%和91.56%,LD50分别为7.16、4.57、5.42 mg/L,以乙酸乙酯提取物的熏杀效果最好。3种提取物对试虫体内的AChE、GSTs活性均表现出明显的剂量效应和较强的时间效应,随着处理剂量的升高,对2种酶活力多数表现为抑制作用增强,而低剂量(1.25 mg/L)的甲醇提取物对AChE具有一定的诱导作用;乙酸乙酯、石油醚提取物在48 h而甲醇提取物在60 h对AChE、GSTs活性抑制作用最强;3种溶剂提取物对AChE、GSTs活性的抑制作用大小均表现为乙酸乙酯>石油醚>甲醇。提取物对酶活性的抑制作用与熏杀效果整体呈正相关,说明AChE、GSTs活性下降可能是造成试虫死亡的重要原因之一。因此,八角茴香具有潜在的杀虫应用研究价值。     

内蒙古产白刺、小果白刺和霸王α-葡萄糖苷酶抑制活性

首次利用体外α-葡萄糖苷酶抑制模型对内蒙古产3种蒺藜科植物的9个提取物进行活性评价,并与阳性对照Acarbose比较,发现3种植物均有抑制α-葡萄糖苷酶活性。其中白刺石油醚提取物对α-葡萄糖苷酶的抑制活性(IC50=81.80 mg/L)最高,其余依次为小果白刺乙酸乙酯提取物(IC50=610.29 mg/L),霸王石油醚(IC50=627.22 mg/L)和乙酸乙酯提取物(IC50=838.40 mg/L),它们的抑制活性远大于阳性对照Acarbose(IC50=1103.01 mg/L)。结果发现,不同植物不同溶剂提取物的α-葡萄糖苷酶抑制活性不同。同一植物不同溶剂提取物相比较,甲醇提取物的α-葡萄糖苷酶抑制活性不及乙酸乙酯和石油醚提取物。     

硬枝树花粗提物的抗氧化作用

采用DPPH和ABTS法对硬枝树花石油醚(30~60)、二氯甲烷、乙酸乙酯、甲醇等4个溶剂依次提取得到的浸膏物,进行抗氧化活性测定试验。结果表明:甲醇提取物,乙酸乙酯和二氯甲烷提取物在0.4~1.0 mg/mL检测浓度范围内对.DPPH的清除效果和质量浓度呈现一定的量效关系。甲醇提取物在1 mg/mL时,清除率高达93.99%,二氯甲烷提取物在1 mg/mL时清除率为92.32%,均高于同质量浓度的BHT。乙酸乙酯和二氯甲烷提取物在0.4~1.0mg/mL检测浓度范围内对ABTS.+自由基清除率与质量浓度呈现一定的量效关系,在1 mg/mL时,二氯甲烷提取物对ABTS.+自由基清除率为65.6%,乙酸乙酯提取物对ABTS.+自由基清除率为48%。     

排风藤内生真菌PFT-2发酵产物抗氧化活性研究北大核心CSCD

采用DPPH·法及Fe3+还原力法对排风藤内生真菌PFT-2提取物的抗氧化活性进行研究,首先测定发酵液乙酸乙酯萃取物和菌丝体甲醇总提物的活性,然后测定菌丝体总提物各极性部位,即石油醚、氯仿、乙酸乙酯、正丁醇,丙酮、甲醇和水层剩余物部位的活性,同时以VC作为阳性对照。结果表明:内生真菌PFT-2菌丝体甲醇总提取物各样品抗氧化活性较强,尤其是甲醇提取物部位、丙酮提取物部位和正丁醇部位;它们对DPPH·的清除作用与Vc相当,其中甲醇提取物部位IC50值为0.023 mg/m L,活性略高于VC,同时以上提取物对Fe3+也具有很强的还原能力。     

拟青霉菌代谢产物对菜粉蝶幼虫的生物活性

淡紫拟青霉(Paecilomyces lilacinus)甲醇萃取物柱层析活性馏分DM6和拟青霉(Paeecilomyces sp.)菌株X1甲醇萃取物柱层析活性馏分X1M5对菜粉蝶Pieris rapae(L.)4龄幼虫具有较强的选择性和非选择性拒食作用,处理后24h选择性AFC50分别为5.8652mg/mL和3.5239mg/mL,非选择性AFC50分别为9.0095mg/mL和14.0234mg/mL。饲喂法、注射法、点滴法测定结果表明,DM6和X1M5对菜粉蝶4龄幼虫具有较强的毒杀活性,其中注射法处理后72h,DM6和X1M5对菜粉蝶4龄幼虫的LD50分别为0.1846μg/头和0.6784μg/头。DM6和X1M5以5.0mg/mL饲喂法处理菜粉蝶3龄幼虫后,试虫生长发育受到强烈的抑制作用,发育抑制率分别为73.34%和63.30%。这些结果对于进一步研究拟青霉菌杀虫活性成分具有重要参考意义。     

排风藤内生真菌PFT-2发酵产物抗氧化活性研究

采用DPPH·法及Fe3+还原力法对排风藤内生真菌PFT-2提取物的抗氧化活性进行研究,首先测定发酵液乙酸乙酯萃取物和菌丝体甲醇总提物的活性,然后测定菌丝体总提物各极性部位,即石油醚、氯仿、乙酸乙酯、正丁醇,丙酮、甲醇和水层剩余物部位的活性,同时以VC作为阳性对照。结果表明:内生真菌PFT-2菌丝体甲醇总提取物各样品抗氧化活性较强,尤其是甲醇提取物部位、丙酮提取物部位和正丁醇部位;它们对DPPH·的清除作用与Vc相当,其中甲醇提取物部位IC50值为0.023 mg/m L,活性略高于VC,同时以上提取物对Fe3+也具有很强的还原能力。     

Susceptibility of some clinical isolates of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> to fractions from the aerial parts of <Emphasis Type="Italic">Leuzea carthamoides</Emphasis>

The antimicrobial activity of the dichloromethane extract from aerial parts of Leuzea carthamoides DC. was tested in vitro against 19 Staphylococcus aureus strains (ATCC 25923, CNCTC Mau 43/60, clinical isolates). The extract was fractionated by column chromatography on silica gel into six fractions (petroleum ether, toluene, dichloromethane, ethyl acetate, methanol and water). The minimum inhibitory concentrations (MICs) of the fractions ranged from 64 to 1024 μg/mL. An ethyl acetate fraction (EA 1) with the widest range of activity inhibited all of the strains with MIC in the range 128–512 μg/mL. This fraction exhibited potent activity against strains which showed associated resistance to oxacillin, ciprofloxacin and erythromycin.     

Secondary metabolites of ponderosa lemon (Citrus pyriformis) and their antioxidant, anti-inflammatory, and cytotoxic activities

Column chromatography of the dichloromethane fraction from an aqueous methanolic extract of fruit peel of Citrus pyriformis Hassk. (Rutaceae) resulted in the isolation of seven compounds including one coumarin (citropten), two limonoids (limonin and deacetylnomilin), and four sterols (stigmasterol, ergosterol, sitosteryl-3-beta-D-glucoside, and sitosteryl-6'-O-acyl-3-beta-D-glucoside). From the ethyl acetate fraction naringin, hesperidin, and neohesperidin were isolated. The dichloromethane extract of the defatted seeds contained three additional compounds, nomilin, ichangin, and cholesterol. The isolated compounds were identified by MS (EI, CI, and ESI), 1H, 13C, and 2D-NMR spectral data. The limonoids were determined qualitatively by LC-ESI/MS resulting in the identification of 11 limonoid aglycones. The total methanolic extract of the peel and the petroleum ether, dichloromethane, and ethyl acetate fractions were screened for their antioxidant and anti-inflammatory activities. The ethyl acetate fraction exhibited a significant scavenging activity for DPPH free radicals (IC50 = 132.3 microg/mL). The petroleum ether fraction inhibited 5-lipoxygenase with IC50 = 30.6 microg/mL indicating potential anti-inflammatory properties. Limonin has a potent cytotoxic effect against COS7 cells [IC50 = (35.0 +/- 6.1) microM] compared with acteoside as a positive control [IC50 = (144.5 +/- 10.96) microM].     

In vitro antimicrobial activity of ethanolic fractions of Cryptolepis sanguinolenta

ABSTRACT: BACKGROUND: Following claims that some plants have antimicrobial activities against infectious microbes, the in vitro antimicrobial activities of different solvent fractions of ethanolic extract of Cryptolepis sanguinolenta were evaluated against eight standard bacteria and clinical isolates. METHODS: The solvent partitioning protocol involving ethanol, petroleum ether, chloroform, ethyl acetate and water, was used to extract various fractions of dried pulverized Cryptolepis sanguinolenta roots. Qualitative phyto-constituents screening was performed on the ethanol extract, chloroform fraction and the water fraction. The Kirby Bauer disk diffusion method was employed to ascertain the antibiogram of the test organisms while the agar diffusion method was used to investigate the antimicrobial properties of the crude plant extracts. The microplate dilution method aided in finding the MICs while the MBCs were obtained by the method of Nester and friends. The SPSS 16.0 version was used to analyze the percentages of inhibitions and bactericidal activities. RESULTS: The phytochemical screening revealed the presence of alkaloids, reducing sugars, polyuronides, anthocyanosides and triterpenes. The ethanol extract inhibited 5 out of 8 (62.5%) of the standard organisms and 6 out of 8 (75%) clinical isolates. The petroleum ether fraction inhibited 4 out of 8 (50%) of the standard microbes and 1 out of 8 (12.5%) clinical isolates. It was also observed that the chloroform fraction inhibited the growth of all the organisms (100%). Average inhibition zones of 14.0 +/- 1.0 mm to 24.67 +/- 0.58 mm was seen in the ethyl acetate fraction which halted the growth of 3 (37.5%) of the standard organisms. Inhibition of 7 (87.5%) of standard strains and 6 (75%) of clinical isolates were observed in the water fraction. The chloroform fraction exhibited bactericidal activity against all the test organisms while the remaining fractions showed varying degrees of bacteriostatic activity. CONCLUSION: The study confirmed that fractions of Cryptolepis sanguinolenta have antimicrobial activity. The chloroform fraction had the highest activity, followed by water, ethanol, petroleum ether and ethyl acetate respectively. Only the chloroform fraction exhibited bactericidal activity and further investigations are needed to ascertain its safety and prospects of drug development.     

侧柏乙醇提取物对21种植物病原真菌的抑菌活性

采用菌丝生长速率法测定了侧柏(Platycladus orientalis)叶、小枝、球果和种子4个不同部位乙醇提取物对21种植物病原真菌的抑菌活性。结果显示:(1)在供试浓度为50g/L(相当于干样)时,侧柏各部位乙醇提取物对4种供试植物病原真菌均具有较好抑制作用,其中侧柏叶提取物的抑菌效果最好,对供试葡萄白腐病菌(Conio-thyrium diplodiella)、葡萄黑痘病菌(Elsinoe ampelina)、番茄绵腐病菌(Phytophthora melongenae)和青霉病菌(Penicilliu mexpansum)的EC50分别为:5.424、3.186、8.913和19.000g/L。(2)侧柏叶乙醇提取物的石油醚萃取物和乙酸乙酯萃取物抑菌活性均较好,在供试浓度为0.5g/L时,石油醚萃取物对苹果腐烂病菌(Valsa mali)和葡萄黑痘病菌(E.ampelina)的抑菌率分别为80.35%和60.23%;乙酸乙酯萃取物对以上2种植物病原菌的抑菌率分别为81.88%和64.06%。结果表明:侧柏叶、小枝、球果和种子乙醇提取物均具有一定抑菌活性,叶乙醇提取物的活性最好,活性成分主要集中在石油醚萃取物和乙酸乙酯萃取物中。     

Activity of Nidus Vespae extract and chemical fractions against Streptococcus mutans biofilms

AIMS: To evaluate the effect of Nidus Vespae extract and chemical fractions on the viability and architecture of Streptococcus mutans biofilms. METHODS AND RESULTS: The raw material was first extracted using 95% ethanol/water. Subsequent fractions were prepared from this extract using cyclohexane/ethyl acetate, petroleum ether/ethyl acetate and chloroform/methanol. The biomass dry weight and total protein of samples treated with Nidus Vespae extract and chemical fractions were significantly less than those treated with the vehicle control (P < 0.05). Biofilms treated with Nidus Vespae also resulted in lower percentage of polysaccharide composition. The pH decrease in the biofilm matrix was retarded by Nidus Vespae compared with the vehicle control. Architecture of biofilms treated with Nidus Vespae was different than in the vehicle control and 0.05% chlorhexidine. CONCLUSIONS: Chloroform/methanol fraction was the most effective treatment. SIGNIFICANCE AND IMPACT OF THE STUDY: The significant antibiofilm activity demonstrated by Nidus Vespae shows it to be a promising source of novel anticariogenic agents.     

树脂吸附法分离瓜果腐霉毒素中的除草活性物质

本试验以对禾本科杂草具有除草活性的瓜果腐霉菌株PAM1为试验材料,研究了X-5、AB-8,H-103和S-8等4种大孔吸附树脂对其除草活性物质的吸附性能,结果表明X-5树脂能够较好地吸附该除草活性物质.选用50%乙醇、95%乙醇、乙酸乙酯和丙酮4种溶剂为供试洗脱溶剂,采用2种洗脱方案,对其最佳洗脱溶剂进行了选择,生长抑制作用和种子萌发试验结果表明,以方案1洗脱为最佳.     

Acaricidal activity of Juglans regia leaf extracts on Tetranychus viennensis and Tetranychus cinnabarinus (Acari: Tetranychidae)

Leaf extracts of the walnut, Juglans regia L., were evaluated under laboratory conditions to determine their acaricidal activity on the mites Tetranychus cinnabarinus (Boisduval) and Tetranychus viennensis Zacher (Acari: Tetranychidae). Extracts had both contact and systemic toxicity to these mites. The four solvents tested for preparing crude extracts were petroleum ether, chloroform, ethyl acetate, and methanol. Methanol was the most efficient solvent, with an extraction rate from 17.06 + 0.80 to 20.27 +/- 0.28%. Petroleum ether was the least effective solvent, with extraction rates from 2.30 +/- 0.13 to 2.71 +/- 0.13%. However, the crude extracts with petroleum ether resulted in the highest mite mortality (79.04 +/- 0.52%) in a slide dip bioassay. Mites mortalities from the concentrated extracts prepared by chloroform, ethyl acetate, methanol, or distilled water were significantly lower than petroleum ether. The mean lethal concentrations (LC50) of the extracts from petroleum ether, chloroform, ethyl acetate, methanol, and distilled water to the two mite species were 0.73 +/- 0.04, 1.66 +/- 0.28, 4.96 +/- 0.35, 7.45 +/- 0.67, and 9.91 +/- 0.32 mg/ml, respectively. After liquid chromatography and thin-layer chromatography, the concentrated extracts of petroleum ether were separated into eight fractions and tested for acaricidal activity. Fraction 6 produced significantly higher mite mortality rates than the other groups, killing approximately 90% of both species.     

斑点野生稻拒食活性组分的分离及其对斜纹夜蛾消化酶活性的影响

为了进一步评估斑点野生稻Oryza punctata的抗虫性,采用液 液分配萃取和硅胶柱层析的方法,从斑点野生稻甲醇提取物中分离获得石油醚、氯仿、乙酸乙酯、正丁醇和水的萃取物,测定了其对斜纹夜蛾Spodoptera litura 3龄幼虫拒食活性。结果显示,氯仿萃取物比其他4种萃取物具有更高的拒食活性,在25 mg/mL的浓度下,24 h和48 h的拒食率分别为55.62%和52.66%。氯仿萃取物经硅胶柱层析后,得到14个组分。比较14个组分的拒食活性,发现组分4和10为主要的活性组分。这两个组分对斜纹夜蛾幼虫中肠脂肪酶和α-淀粉酶的活性都具有一定的抑制活性,其中组分10对脂肪酶具有显著的抑制效果,以1 mg/mL的浓度活体处理48 h和72 h抑制率分别达19.82％和34.60％; 对α-淀粉酶的影响在48 h和72 h内都具有显著的抑制效果,随着处理时间的延长,其抑制率逐步提高,72 h的抑制率分别为25.06％和27.40％。结果提示斜纹夜蛾幼虫中肠脂肪酶和α-淀粉酶可能是斑点野生稻拒食活性成分的作用靶标。     

乳孔硫磺菌的化学成分和抗氧化活性

对乳孔硫磺菌子实体不同极性提取物进行了DPPH和ABTS自由基清除能力的测定,并对氯仿和乙酸乙酯提取物进行了分离纯化。结果表明,乳孔硫磺菌的石油醚、氯仿、乙酸乙酯和甲醇提取物均有一定的抗氧化活性,各提取物对两种自由基的清除能力均表现为甲醇提取物＞乙酸乙酯提取物＞氯仿提取物＞石油醚提取物,甲醇提取物对DPPH自由基的清除率最高可达到93.78%;对ABTS自由基的清除率最高可达到62.06%;从氯仿和乙酸乙酯提取物中分离得到10个化合物,分别是：(22E,24R)-5α,8α-过氧麦角甾-6,22-二烯-3β-醇（1）,阿里红酸 A（2）,麦角甾-7,22-二烯-3β-醇（3）,啤酒甾醇（4）,硫色多孔菌酸（5）,(4E,8E)-N-d-2′-hydroxypalmitoyl-1-O-β-d-glycopyranosyl- 9-methyl-4,8-sphingadienine（6）,麦角甾醇（7）,N-(2′-羟基二十四碳酰基)-1,3,4-三羟基-2-氨基-十八烷（8）,烟酸（9）和齿孔酸（10）。其中,化合物2、6、8和9为首次从硫磺菌属真菌中分离得到。     

小珊瑚藻对赤潮异弯藻的化感效应

研究了不同浓度的小珊瑚藻组织4种不同极性有机溶剂(甲醇、丙酮、乙醚、氯仿)提取物对赤潮异弯藻的生长抑制作用.结果表明：小珊瑚藻组织甲醇提取物对赤潮异弯藻的生长抑制活性最强,并且在较高浓度下能使赤潮异弯藻完全死亡,其他3种有机溶剂提取物对赤潮异弯藻的生长无明显影响.表明小珊瑚藻组织中含有的对赤潮异弯藻有抑制作用的活性物质具有较高的极性.对小珊瑚藻的甲醇提取物进行液液萃取,将其分离为石油醚相、乙酸乙酯相、正丁醇相和蒸馏水相, 并对赤潮异弯藻进行生物活性检测.结果发现石油醚相和乙酸乙酯相具有较强的杀藻活性,表明脂肪酸可能是小珊瑚藻组织内抑制赤潮异弯藻生长的化感物质的重要组成成分之一.