柱层析分离纯化功能性红曲米中的有效成分

为深入研究功能性红曲米浓缩物中有效成分的性质及其功能,采用硅胶柱层析法(依次用乙酸乙酯:石油醚(1∶1,v∶v)、乙酸乙酯、无水甲醇、蒸馏水进行洗脱)对功能性红曲米浓缩物中的活性成分进行分离纯化,得到红曲黄色素、洛伐他汀粗品、醇溶性红曲红色素、水溶性红曲红色素4种组分;并采用紫外-可见扫描和高效液相色谱法对4种组分进行简单的性质结构表征.     

山茱萸的化学成分及其抗氧化活性

为明确药食两用山茱萸化学成分的抗氧化活性,本研究以山茱萸乙醇提取物为原料,对乙醇提取物大孔吸附树脂20%乙醇洗脱部位中抗氧化活性组分进行研究,采用大孔吸附树脂柱层析常压正相硅胶柱层析、Sephadex LH-20凝胶柱层析、Flash闪式正反相硅胶柱层析、高压液相色谱等色谱方法对山茱萸乙醇提取物抗氧化活性部位进行系统的分离纯化,并结合核磁共振波谱数据、质谱数据与文献对比,从山茱萸乙醇提取物中共分离鉴定了5个化合物,分别为7-β-O-乙基莫诺苷(1)、7-α-O-乙基莫诺苷(2)、3-O-咖啡酰奎宁酸甲酯(3)、(7R, 9R, 10R)-3, 9-di-hidroxicalameneno(4)和吐叶醇(5);其中化合物3、4和5为首次从山茱萸中分离得到。同时采用1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)法和铁离子还原法(Ferric ion reducing antioxidant power,FRAP)对分离得到的单体化合物进行抗氧化活性筛选。在DPPH自由基清除试验和FRAP试验中,化合物1、2和3均显示一定的抗氧化能力,其中化合物3抗氧化活性最好。     

1 株具抑菌和抗氧化活性乳酸菌的筛选及鉴定

从市售酸泡菜中分离纯化乳酸菌,以植物乳杆菌PL2、大肠杆菌As1.184等为指示菌筛选具有抑菌活性的乳酸菌;以对1,1-二苯基-2-苦基肼（1,1-diphenyl-2-picrylhydrazyl,DPPH）自由基、·OH、O2－·清除率为指标,进一步筛选高抗氧化活性菌株。结果表明：从市售酸菜中分离筛选出8?株兼具抑菌活性和抗氧化活性的乳酸菌菌株,其中菌株b-2的抑菌活性和抗氧化活性最强,根据生理、生化和分子生物学特征将其鉴定为植物乳杆菌。抗氧化活性测定结果显示,该菌株的无细胞提取物对·OH的清除率最高,为67.6%;其完整细胞对O2－·的清除活性最强,清除率为62.8%;其无细胞发酵上清液对DPPH自由基清除率高达91.3%。植物乳杆菌b-2具有较好的抑菌能力和抗氧化能力,作为功能性乳酸菌发酵剂,具有十分重要的开发潜力。     

杜仲雄花乙酸乙酯提取物的抑菌活性研究

本实验以杜仲雄花为材料,通过打孔法和倍比稀释法,探讨杜仲雄花乙酸乙酯提取物及经过硅胶柱分离得到的六种不同极性提取物对真菌和细菌的抑菌活性,并确定最小抑菌浓度(MIC)。结果显示:杜仲雄花乙酸乙酯提取物对供试金黄色葡萄球菌、炭疽杆菌、枯草芽孢杆菌、大肠杆菌和黑曲霉均有较好的抑制效果,最小抑菌浓度(MIC)分别为0.0156、0.0156、0.0313、0.0313、0.1250g/m L,对根霉和青霉没有抑制作用。而分离得到的六种提取物按极性不同分别为10%甲醇洗脱所得样品、20%甲醇洗脱所得样品、30%甲醇洗脱所得样品、纯甲醇洗脱所得样品、纯甲醇含氨洗脱所得样品和水洗脱所得样品,其中10%甲醇洗脱所得样品、20%甲醇洗脱所得样品与乙酸乙酯提取物活性相似或有增强,30%甲醇洗脱所得样品、纯甲醇洗脱所得样品对部分细菌有抑制作用,而只有水洗脱所得样品对根霉有抑菌作用。说明杜仲雄花乙酸乙酯提取物和进一步的硅胶柱分离得到的不同极性提取物具有良好的抑菌活性,且硅胶柱的分离能得到更佳的抑菌活性提取物,为杜仲雄花的进一步开发提供依据。     

微拟球藻活性成分分离及抑菌活性研究

海洋微藻因其特殊的生存环境使其可以合成高附加值的代谢产物而成为天然抗菌素筛选的重要途径。采用纸片法对微拟球藻的不同极性提取物及其分离组分进行抗4种病原菌活性研究,并采用柱层析和薄层层析方法对石油醚提取物进行进一步的分离纯化及成分分析,结果显示:6种提取物在较高的受试浓度下对大肠杆菌都呈现出较强的抑菌活性,此外,乙醇相提取物对其它3种受试菌都表现出明显的抑制效果,而石油醚相对铜绿假单胞菌有一定的抑菌活性,乙酸乙酯相和正丁醇相提取物对白色念珠菌有明显的抑菌效果。石油醚相提取物进一步分离获得3个纯度较高的组分,成分分析结果显示PEⅢ-5-1为叶绿素类化合物;PEⅤ-1中酮类物质含量最多,占55.74%,其次是酯类;PEⅤ-2中含量最多的是酯类,占50.54%;其次是酮类和有机酸类。     

苦丁茶提取物的分离纯化、鉴定及不同萃取部位活性分析

以苦丁茶为原料，对其乙醇提取物和不同溶剂萃取部位的抗氧化活性、抗补体活性、抑制癌细胞活性进行探究，并对活性最佳的萃取部位进一步分离纯化，分析鉴定出单体化合物。抗氧化活性以总还原力、清除1,1-二苯基-2-三硝基苯肼自由基和超氧阴离子自由基的能力为指标，通过溶血实验评价抗补体活性，并采用MTT法测定对人乳腺癌细胞MCF-7的存活率，结合多种分离技术获得不同单体化合物，通过对质谱、核磁共振波谱的数据分析鉴定其化学结构。结果表明，苦丁茶乙醇提取物及各部位均具有良好的抗氧化效果，其中乙酸乙酯部位不仅在实验质量浓度范围内有较强的抗氧化活性（0.8 mg/mL均达到90%以上），还具有良好的抗补体活性（79.67±0.99）%，能很好抑制人乳腺癌MCF-7细胞增殖活性（54.8±0.26）%，对乙酸乙酯部位进行分离纯化，总共分离得到11 个单体化合物。由此可见，苦丁茶乙醇提取物和各部位均具有抗氧化、抗补体活性和人乳腺癌MCF-7细胞抑制活性，乙酸乙酯作为萃取剂时活性最佳。分离的单体化合物鉴定后多为酚类、黄酮类物质。     

粗毛纤孔菌(Inonotus hispidus)的鉴定及其子实体不同溶剂提取物的抗氧化活性与抑菌活性研究

采用rDNA ITS序列分析对一株野生桑黄菌(SH)进行鉴定,研究了其子实体不同溶剂(70%乙醇、甲醇、乙酸乙酯、氯仿)提取物中的总黄酮、总多酚与抗氧化活性的相关性以及4种提取物的抑菌活性。结果表明:该野生桑黄菌为粗毛纤孔菌(Inonotus hispidus),其子实体不同溶剂提取物的总黄酮、总多酚、抗氧化活性和抑菌活性存在显著差异,其中甲醇提取物中的总黄酮和总多酚含量最高,分别为(14.78±0.16)mg芦丁/g和(67.42±1.41)mg没食子酸/g;70%乙醇提取物的和DPPH·清除能力ABTS+·清除能力最强,分别为(43.83±0.03)和(116.08±0.24)mg Trolox/g,甲醇提取物的铁离子还原能力最强,为(126.24±3.02)mg Trolox/g;总多酚和总黄酮含量与FRAP法之间呈极显著正相关(p<0.01),总多酚与ABTS+·清除能力和DPPH·清除能力之间呈显著相关(p<0.05),而总黄酮与ABTS+·清除能力和DPPH·清除能力不相关;70%乙醇提取物和甲醇提取物的抑菌效果最好,其中对草莓灰葡萄孢霉的抑菌效果最好(抑菌圈直径>30 mm),其次是对金黄色葡萄球菌(抑菌圈直径>15 mm),对大肠杆菌的抑菌效果最差(抑菌圈直径>10 mm)。研究发现粗毛纤孔菌甲醇提取物的总多酚、总黄酮含量最高,抗氧化活性和抑菌活性最强,是天然抗氧化剂的良好来源。     

质谱图筛选红曲红色素纯化方法

以商品化的红曲米为原料,利用质谱图定性分析,考察在红曲红色素提取过程中冷冻,索氏提取,溶剂,柱色谱,薄层色谱等因素对分离效果的影响,筛选并分离得到一种分子离子峰为332.6红曲红色素分子。研究结果表明该红曲红色素分子的纯化条件依次为:以甲醇为淋洗剂对红曲米进行柱色谱分离,以正己烷、乙酸乙酯和甲醇为萃取剂进行一次索氏提取,以CH2Cl2∶CH3OH=1∶1为展开剂进行薄层色谱分离,收集Rf=0.8的色带,以CH3CH2OH∶CCl4=9∶1为展开剂进行第二次薄层色谱分离,收集Rf=0.8的色带为目标分子,收率为0.012 6%。     

石榴皮多酚超高压提取物的纯化、分离及功能性研究

为探明超高压提取石榴皮多酚的组分和功能特性,研究了石榴皮多酚超高压提取物的大孔树脂纯化和有机溶剂分级萃取,并对提取物、纯化物和分级产物的体外抗氧化活性和抑菌性能进行了测定。结果表明:大孔树脂纯化物的总酚含量高达77.40%,且抗氧化活性和抑菌性能均有显著提高;HPLC测定结果显示,大孔树脂纯化未改变多酚种类;有机溶剂分级萃取法能有效地将纯化物进一步分离,其中正丁醇相的萃取得率达到55.28%;不同萃取相产物的总酚含量大小顺序为:正丁醇相>乙酸乙酯相>水相,它们的抗氧化活性强弱顺序为:乙酸乙酯相>正丁醇相>水相;正丁醇相产物的抑菌活性最高,显著高于水相和乙酸乙酯相。     

排草提取物抗氧化和抑菌活性的研究

为了研究排草的抗氧化和抑菌活性,以水作为溶剂得到排草水提取物,分别采用DPPH法、ABTS法和还原力法测定其抗氧化活性。同时进行体外抑菌试验,测定排草水提取物对大肠杆菌、枯草芽孢杆菌、金黄色葡萄球菌、苏云金芽孢杆菌、肠球菌等5种常见食源性污染菌的抑菌效果。结果表明:排草水提取物具有一定的抗氧化活性,对ABTS自由基的清除效果最好,EC50为22.71μg/mL。体外抑菌试验表明排草水提取物对肠球菌和苏云金芽孢杆菌有很好的抑菌效果,最低抑菌浓度为60mg/mL,抑菌强弱顺序为:肠球菌>苏云金芽孢杆菌>枯草芽孢杆菌>金黄色葡萄球菌>大肠杆菌。     

杜仲雄花正丁醇提取物抑菌活性研究

为探讨杜仲雄花正丁醇提取物的抑菌活性,以大肠杆菌、枯草芽孢杆菌、金黄色葡萄球菌、炭疽杆菌、根霉、青霉和黑曲霉为供试菌种,采用打孔法和倍比稀释法,检测杜仲雄花正丁醇粗提物及经过硅胶柱分离得到的7种不同极性提取物对真菌和细菌的抑菌活性,并确定最小抑菌浓度(MIC),分析粗提物及7种分离产物的活性。结果显示,杜仲雄花正丁醇粗取物对金黄色葡萄球菌和炭疽杆菌具有较好的抑菌作用,MIC分别为0.007 8g/mL和0.015 6g/mL;杜仲雄花正丁醇粗提物对枯草芽孢杆菌、大肠杆菌、根霉、黑曲霉和青霉没有抑制作用。然而经过硅胶柱进一步分离发现,经过分级提取的样品4和样品5对黑曲霉出现了抑制作用,对金黄色葡萄球菌、炭疽杆菌、和枯草芽孢杆菌等革兰氏阳性菌的抑制作用明显增强。说明杜仲雄花正丁醇粗提物经过硅胶柱的分离能够得到对不同菌株的特异抑菌物质,这为寻找不同菌种的抑菌活性物质提供了参考依据。     

解淀粉芽孢杆菌抑菌物质粗提取的优化及分析

为探究解淀粉芽孢杆菌（Bacillus amyloliquefaciens）A13抑菌物质适宜提取方法,分别采用硫酸铵盐析法、酸沉淀法、甲醇抽提法、乙酸乙酯萃取法进行抑菌粗提液的制备,并通过SuperdexTM75凝胶层析柱分离纯化。结果表明,以80%饱和硫酸铵法制备粗提液抑菌效果最佳,抑菌圈直径为23.67 mm,蛋白质量浓度最高,为582.50 μg/mL,其回收率为17.8%;以甲醇抽提法制备粗提液的抑菌圈直径为25.17 mm,其回收率最高,为32.1%,蛋白质量浓度为505.00 μg/mL;以酸沉淀法制备粗提液的抑菌圈直径为22.67 mm,其回收率与蛋白质量浓度分别为13.7%与273.75 μg/mL;以乙酸乙酯法制备粗提液时,乙酸乙酯与发酵液为3∶1（V/V）的抑菌效果最佳,抑菌圈直径为25.5 mm,并且其回收率和蛋白质量浓度分别为23.6%和255.20 μg/mL。因此,硫酸铵盐析法为粗提解淀粉芽孢杆菌A13抑菌物质的适宜方法。     

In vitro antioxidant/free radical scavenging and antibacterial properties of endemic oregano and thyme extracts from Greece

The present investigation was to evaluate the antioxidant and antibacterial activities of hexane and methanol extracts from Origanum vulgare ssp. hirtum and Thymus vulgaris (Komotini, Greece). The methanol extracts of oregano and thyme against DPPH radical were more active than the hexane extracts (mean values 0.94 and 0.47, respectively) and oregano exhibited stronger activity than thyme (mean values 0.82 and 0.55, respectively). The results from the β-carotene/linoleic acid assay showed that all plant extracts inhibited linoleic acid oxidation up to 70.78±1.17%. The oregano extracts exhibited the strongest inhibition against Escherichia coli, Listeria monocytogenes, and Staphylococcus aureus using the disc diffusion assay. The chemical composition of the hexane and methanol extracts, by using GC-MS, showed that carvacrol, thymol, and p-cymene were the most prominent compounds. The methanol extract of oregano was found the most potent antioxidant with the highest content of total phenolics (138.92 mg GA/g extract) and carvacrol (76.7%).     

Chemical Composition,Antioxidant, and Antibacterial Activities of the Essential Oil and Methanol Extracts of Eucalyptus oleosa Leaves

This study was designed to examine the chemical composition of essential oil and in vitro antioxidant and antibacterial activity of the essential oil and methanol extracts of Eucalyptus oleosa F. Muell. The chemical composition of the hydrodistilled essential oil of the leaves of E. oleosa was analyzed by gas chromatography and gas chromatography/mass spectrometry. The main constituents of the oil were found to be 1,8-cineole (45.1%), α-pinene (14.52%), and α-terpineol (4.35%). The essential oil showed strong antibacterial activity against the test microorganisms studied, while polar subfraction of methanol extract had moderate antibacterial activity and the nonpolar subfraction of methanol extract did not show any antibacterial activity. In contrast, the extract showed much better activity than the essential oil in antioxidant activity assays employed, e.g., in DPPH systems, the highest radical-scavenging activity was shown by the polar subfraction (15.1 ± 0.7 μg/ml). In the second case, the inhibition capacity (%) of the nonpolar subfraction (98.2% ± 1.5) was found to be the stronger one. In addition, the amounts of total phenol components in the polar subfraction (186.3 ± 2.1 μg/mg) and the nonpolar subfraction (79.6 ± 1.4) were determined.     

Antioxidant and antimicrobial activities of the methanol extracts from pummelo (Citrus grandis Osbeck) fruit albedo tissues

The present study was conducted to evaluate the antioxidant and antimicrobial properties of methanol (100 and 80% aqueous) extracts of pummelo fruits albedo (Citrus grandis Osbeck). The antioxidant and antibacterial activity for crude extracts and isolated compounds were evaluated using free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) and Paper disc diffusion method. A 100% Methanol (MeOH) extract was steeped in water at different pH levels and partitioned with ethyl acetate (EtOAc) to give basic, acidic, neutral, and phenolic fractions. The neutral extract was found to possess maximum antioxidant and antibacterial activity. Thereafter, neutral extract was carried out on a silica gel column and eluted with hexane:EtOAc:acetone and preparative TLC (PTLC) to give oil buntan compound, linoleic acid methyl ester, β-sitosterol, sigmasterol, limonin, nomilin and meranzin hydrate were isolated. While, 80% MeOH extract was fractionated also using a silica gel column and PTLC to give isomeranzin hydrate, p-coumaric acid and caffeic acid compound. The extract concentration providing 50% inhibition (IC₅₀) was as follows; oil buntan compound 95 μg/mL, caffeic acid 45 μg/mL, p-coumaric acid 105 μg/mL, limonin + nomilin (mixture) 135 μg/mL was lower than that of synthetic antioxidant butylated hydroxyanisole (BHA) 40 μg/mL. The inhibitory zone (mm) of bacteria tested was 2.9–4.1 mm caffeic acid and 11.6–15.1 mm p-coumaric acid.     

荸荠皮抑菌活性成分的研究

目的 鉴定荸荠皮中抑菌活性成分的种类及结构.方法 先将荸荠皮粗提物用不同极性的有机溶剂分步提取,通过抑菌实验筛选抑菌活性最强的提取物,通过硅胶柱色谱分离纯化,得到抑菌活性最强的两个部分,气相色谱-质谱（GC-MS）鉴定抑菌物质的种类及结构.结果 荸荠皮抑菌物质中含有甾醇类、有机酸类、酯类和黄酮类4类活性成分共11种活性物质.结论 此鉴定结果符合荸荠皮提取物的抑菌特性,为荸荠皮抑菌物质的研究提供了依据.     

Evaluation of the antioxidant potential of Pittosporum dasycaulon Miq. stem bark

The in vitro antioxidant properties of methanol and aqueous extracts of stem bark from Pittosporum dasycaulon Miq. were investigated. Fractions were screened for total antioxidant capacity, ferric reducing power, and radical scavenging activities. The total phenolic, flavonoid, and tannin contents of the methanol and aqueous extracts were also determined. The aqueous extract exhibited a stronger scavenging effect than the methanol extract against the DPPH free radical, the nitric oxide radical, and the superoxide radical. The methanol extract exhibited a stronger total antioxidant activity and reducing power than the aqueous extract. The free radical scavenging effect of P. dasycaulon stem bark extracts was comparable to an ascorbic acid reference antioxidant while the reducing power of the methanol extract was greater than ascorbic acid. P. dasycaulon stem bark is a potential source of natural antioxidant compounds.     

菱角梗、菱角壳提取物抑菌活性及其活性成分分析

采用滤纸片扩散法研究菱角梗、菱角壳乙醇提取液及其不同溶剂萃取物的抑菌活性,并考察不同温度、pH、紫外辐照对菱角梗、菱角壳乙酸乙酯萃取物抑菌活性稳定性的影响,最后,采用高效液相色谱法和超高效液质联用法对菱角梗、菱角壳乙酸乙酯萃取物中的成分进行鉴定。结果表明:菱角梗、菱角壳乙酸乙酯萃取物对金黄色葡萄球菌、大肠杆菌和酿酒酵母菌的最小抑菌浓度分别为0.391、0.391、3.125 mg/mL和0.098、0.195、1.563 mg/mL,抑菌效果显示,菱角壳乙酸乙酯萃取物优于菱角梗乙酸乙酯萃取物,二者萃取物均对革兰氏阳性菌具有显著抑菌作用,且效果强于革兰氏阴性菌,对真菌的抑菌作用最弱;菱角梗、菱角壳乙酸乙酯萃取物在温度20~100℃、pH5~9和紫外光处理10~40 min时,均表现出良好的抑菌稳定性。经鉴定,菱角梗乙酸乙酯萃取物中含量最高的是没食子酸和1,2,6-三-O-没食子酰-β-D-葡萄糖,含量为45.87%,菱角壳乙酸乙酯萃取物中含量最高的是1,2,3,6-四-O-没食子酰-β-D-葡萄糖、槲皮素-3-O-半乳糖苷和1,2,6-三-O-没食子酰-β-D-葡萄糖,含量为57.84%。本文为菱角梗、菱角壳提取物作为抑菌剂的实际应用提供了参考。     

油菜蜂花粉酚类化合物的初步分离和抗氧化性研究

采用甲醇提取油菜蜂花粉,提取物再经正丁醇萃取后,用聚酰胺层析得到各分离组分,并对各分离组分进行抗氧化性指标测定,并进行TLC、HPLC 分析。结果表明：甲醇提取物酚类化合物含量为3.69%,正丁醇萃取物为6.32%,聚酰胺层析中80% 和100% 乙醇洗脱组分酚类化合物含量分别为15.92% 和16.87%。同时,测定蜂花粉中各分离组分在浓度为1mg/ml 时的还原力和DPPH·清除率,甲醇提取物还原力为26μgVC/mg,正丁醇萃取物为84μgVC/mg,80%和100%乙醇洗脱组分提高到154μgVC/mg 和104μgVC/mg;甲醇提取物DPPH·清除率为83.45%,正丁醇萃取物为88.66%,聚酰胺层析中80% 和100% 乙醇洗脱组分DPPH·清除率分别为97.20% 和94.34%。实验表明：蜂花粉经过甲醇提取、正丁醇萃取和聚酰胺层析后,80% 乙醇和100% 乙醇分离组分酚类化合物含量和抗氧化活性均显著性提高。     

<Emphasis Type="Italic">Borago officinalis</Emphasis> L. flower: a comprehensive study on bioactive compounds and its health-promoting properties

The herbal plant Borago officinalis L. flower were analysed for its bioactive compounds and antioxidant, antibacterial, anti-inflammatory and anticancer activities using different solvent polarities (methanol, ethanol and water). The RP-HPLC analyses of the methanolic extract confirmed the presence of phenolics (gallic acid, pyrogallol, salicylic acid, caffeic acid), flavonoids (myricetin, rutin) and isoflavonoid (daidzein). Besides, the major individual fatty acids of methanolic extract were α-linolenic, stearidonic, palmitic, linoleic and γ-linolenic acids. The methanolic extract possessed the highest antioxidant properties as compared to the ethanolic and water extracts. The flower methanolic, ethanolic and water extracts showed high, moderate and weak antibacterial activities against common human and foodborne pathogenic bacteria, respectively. Furthermore, the flower extracts showed weak anti-inflammatory in murine RAW 264.7 macrophage cells and low anticancer properties against human hepatic, prostate and colon cancer cells. This high-value flower could be considered as a source of putative antioxidant and antibacterial compounds to improve the human health and to be used as biopreservative in food and cosmetic industries.