保加利亚乳杆菌对绞股蓝皂苷Gypenoside XLⅥ的微生物转化

目的：分析绞股蓝皂苷Gypenoside ⅩLⅥ的微生物转化产物。方法：利用益生菌德氏乳杆菌保加利亚亚种的脱脂牛奶培养基对Gypenoside ⅩLⅥ进行微生物转化,并利用液相色谱离子阱飞行时间串联质谱方法鉴定Gypenoside ⅩLⅥ的微生物转化产物。结果：Gypenoside ⅩLⅥ的微生物转化产物为绞股蓝皂苷Gypenoside L、Gypenoside LI、Damulin B 和Damulin A。结论：通过微生物转化方法可以获得更多天然产物活性成分。     

绞股蓝皂苷成分的分离与鉴定

目的 提取、分离、鉴定绞股蓝皂苷主要成分。方法 利用硅胶柱色谱及反相半制备柱色谱等分离手段, 分离、纯化绞股蓝皂苷成分, 并用核磁共振波谱(1H-NMR、13C-NMR)、液相色谱离子阱飞行时间质谱(LCMS-IT-TOF)等方法鉴定化合物结构, 并对相关成分的抑癌作用进行了研究。结果 绞股蓝总提物的主要成分经分离得到2种达玛烷类皂苷, 分别为绞股蓝皂苷gypenoside XLVI和gypenoside LVI, 而且它们具有一定的抑制癌细胞增殖作用。结论 利用硅胶柱色谱和反相半制备色谱方法从绞股蓝总提物中分离得到主要的皂苷成分, 为进一步研究其抑癌有效成分提供了参考。     

福建绞股蓝皂苷的优化提取及其抗肝癌活性

绞股蓝素有"南方人参"美誉,含有与人参皂苷相同骨架达玛烷型结构的三萜皂苷。通过不同体积分数乙醇溶液优化提取绞股蓝皂苷,测定其抗癌活性,为后续活性化合物的分离奠定基础。利用HPLC分析绞股蓝的皂苷成分并确定提取绞股蓝皂苷的乙醇体积分数,正交实验优化提取方法,进而MTT法测定提取的绞股蓝皂苷抗肝癌活性。HPLC分析确定用体积分数75%乙醇溶液提取绞股蓝总皂苷(GYP),体积分数45%乙醇提取极性绞股蓝皂苷(GYP1),体积分数90%乙醇提取弱极性绞股蓝皂苷(GYP2);正交实验结果表明在70℃水浴中以30质量的体积分数45%乙醇加热提取12 h为最佳条件提取GYP1,在60℃水浴中以30质量的体积分数90%乙醇加热提取6 h为最佳条件提取GYP2;利用MTT法测定GYP、GYP1和GYP2对肝癌细胞的抑制作用,GYP和GYP2对肝癌细胞SMCC7721具有抗癌活性,半抑制率IC50值分别为(165.83±14.12)μg/mL和(113.97±9.26)μg/mL,GYP1没有抗癌活性,表明绞股蓝抗癌药效成分主要是由弱极性绞股蓝皂苷发挥功效。     

壮药“国虾薄”(绞股蓝)热处理产物抗肺癌的有效成分研究进展

细胞凋亡通路受阻是导致肿瘤发生、发展及产生耐药性的主要原因, 通过死亡受体信号转导途径和线粒体途径诱导细胞凋亡, 是近年来发现的抗肿瘤治疗的主要机制。壮药“国虾薄”通三道两路, 具有清热解毒的作用其热处理产物具有很强的抑制非小细胞肺癌(NSCLC)A549细胞增殖活性, 并诱导细胞凋亡。但其有效成分尚不明确, 作用机制尚不清楚。本文介绍了细胞凋亡通路、壮医药抗肿瘤基本理论、热处理方法在天然药物中的应用、壮药“国虾薄”(绞股蓝)的生物活性及其热处理产物抗NSCLC有效成分研究。     

蔬菜种子中5种异硫代氰酸酯类化合物对人肺癌细胞抑制作用的研究

以人肺癌A549细胞为研究对象,采用MTT法检测5种异硫代氰酸酯对A549细胞生长的抑制作用,计算其IC50值。采用倒置显微镜观察细胞形态变化。通过研究5种异硫代氰酸酯对A549细胞的影响,探讨这些化合物的抗癌作用及其结构与活性之间关系。结果表明,5种单体对肿瘤细胞均有不同程度的剂量依赖性抑制作用,并且能使A549细胞的形态发生改变;对A549细胞的抑制活性是:萝卜子素萝卜硫素≈3-(甲磺酰基)丙基异硫代氰酸酯3-(甲硫基)丙基异硫代氰酸酯3-丁烯基异硫代氰酸酯,IC50值分别为(15.43±0.82)μmol/L,(22.09±0.94)μmol/L,(29.82±1.36)μmol/L,(47.40±3.51)μmol/L,(79.57±3.05)μmol/L。萝卜子素的有效浓度为3.125～100μmol/L;萝卜硫素、3-(甲磺酰基)丙基异硫代氰酸酯、3-(甲硫基)丙基异硫代氰酸酯、3-丁烯基异硫代氰酸酯的有效浓度均为6.25～100μmol/L。说明异硫代氰酸酯的特征基团-N=C=S为主要活性基团,侧链上的磺酰基和硫基可能是异硫代氰酸酯的活性辅助位点。侧链上含有磺酰基的异硫代氰酸酯类化合物具有较强的抑制肿瘤活性。     

提取方法对木槿叶挥发油成分及其对肺癌A549细胞抑制作用的影响

分别采用超临界CO_2萃取(SFE—CO_2)和水蒸气蒸馏法(SDE)提取木槿叶中挥发油,利用气相色谱—质谱(GC—MS)鉴定化合物,检测细胞凋亡指数和凋亡率评定挥发油抑制肺癌A549细胞作用。从SFE—CO_2挥发油中鉴定40个化合物,从SDE挥发油中鉴定47个化合物。结果表明:木槿叶SFE—CO_2挥发油与SDE挥发油对肺癌A549细胞抑制率(浓度25mg/L,作用48h)分别为80.08%,62.75%,说明不同方法提取的挥发油抗肿瘤活性差异显著。验证实验表明,雌甾-1,3,5(10)-三烯-17-醇、1-己基-6-羟基-4-甲基六氢嘧啶-2-硫酮、6,6-二甲基-9-(3-环氧丙烷-2-基)-1,4-二氧杂螺[4.5]癸烷、2,4-二叔丁基苯酚、丁香酚是抑制肺癌细胞A549的主要活性成分。     

绞股蓝皂苷生物转化与活性的研究进展

绞股蓝为葫芦科绞股蓝属植物,含皂苷、黄酮、多糖等多种成分,具有防衰老、降血糖、降血脂、抑制肿 瘤等多种生物活性。绞股蓝皂苷是绞股蓝的主要活性成分之一,因其具有与人参皂苷相同的四环三萜达玛烷型母核 结构而备受关注。近年来,通过生物转化对绞股蓝皂苷上的糖基进行改造以制备活性较高的次生皂苷成为研究热 点,并出现大量相关的研究报道,但鲜有较为系统的文献整理。鉴于此,本文综述了绞股蓝皂苷的结构与类别、绞 股蓝皂苷与人参皂苷的异同、绞股蓝皂苷的生物转化及其生物活性的研究进展,并就绞股蓝皂苷未来的研究方向提 出建议,以期为绞股蓝皂苷的进一步研究提供思路。     

高效液相色谱法测定绞股蓝提取物中的皂苷成分

目的建立绞股蓝提取物皂苷成分的高效液相色谱测定法,并测定市场上绞股蓝提取物皂苷含量。方法比较甲醇、乙醇、水做提取溶剂及超声、回流提取方式下的皂苷得率。色谱柱为Waters Synnetry C_(18)(4.6 mm×250 mm,5μm);柱温:35℃;流速1.0 m L/min;检测波长为203 nm。结果以甲醇为溶剂,超声30 min皂苷含量较高。绞股蓝皂苷XLⅨ、绞股蓝皂苷XLⅥ、绞股蓝皂苷A、皂苷XⅦ分别在2.605~130.24μg/mL(r~2=0.9992)、2.205~110.24μg/mL(r~2=0.9995)、1.981~99.04μg/mL(r~2=0.9992)、1.794~89.68μg/m L(r~2=0.9995)范围内线性关系良好,平均回收率分别为98.5%~100.4%、97.3%~101.4%、98.0%~101.1%及99.1%~100.6%。结论本方法灵敏、准确、专属性强,能准确有效地检测绞股蓝提取物中4种皂苷的含量。     

盐生海芦笋来源真菌Salcoli6发酵产物抗肿瘤活性成分研究

采用活性追踪的方法,对一株盐生海芦笋来源真菌Salcoli6发酵产物中的抗肿瘤活性成分进行分离和鉴定。利用溶剂萃取、硅胶柱色谱及制备HPLC等分离手段从发酵产物中分离得到7个化合物,通过理化性质分析及波谱学方法鉴定其化学结构分别为枝孢菌素(1)、异枝孢菌素(2)、2-乙酰基喹唑啉(3)、萘满酮(4)、3-羟基-2-甲级色原酮(5)、3-甲级异苯并呋喃(6)、对-羟基苯乙酸(7);以SRB法评价化合物的抗肿瘤活性,化合物3、4和7对p388细胞具有强的细胞毒活性,IC50值分别为12、24μg/mL和6μg/mL,其他化合物没有活性。     

多穗柯挥发性组分分析及抗癌活性的初步研究

本研究以多穗柯为材料,采用同时蒸馏萃取法(SDE)提取挥发性组分,通过气质联用仪(GC-MS)分析挥发性组分组成,并以Hepa1c1c7小鼠肝癌细胞模型,以细胞存活率大于50%,诱导醌还原酶(QR)活性等于或大于2为活性指标,研究多穗柯挥发性组分及其主要单体物质的抗癌活性。结果显示,GC-MS共鉴定出80种多穗柯挥发性物质,占总挥发性组分相对百分含量的92.56%,其中酮类、醇类和醛类物质是多穗柯挥发性组分的主要类型,其主要成分有香叶基丙酮、β-紫罗酮、壬醛和桉油烯醇等。抗癌活性鉴定表明,多穗柯挥发性组分的浓度为(27.50±0.10)μg/mL时可诱导QR活性达到倍增,具有较强的抗癌活性;其中芳樟醇、香叶基丙酮和壬醛三个主要挥发性单体成分诱导QR活性达到倍增的浓度分别为(54.53±0.10)μg/mL、(283.10±0.10)μg/mL和(297.77±0.12)μg/mL。     

绞股蓝皂苷微丸的制备工艺

研究绞股蓝皂苷微丸的制备工艺。以挤出频率、滚圆频率、滚圆时间为考察因素,以微丸的圆整度为评价指标,利用单因素法研究制备微丸的最优工艺条件。采用挤出滚圆法制备空白微丸;采用流化床法制备绞股蓝皂苷微丸。并对微丸的粉体学性质和体外释放度进行测定;用红外光谱法、X-射线粉末衍射法、扫描电子显微镜法对其进行表征。结果显示,制备微丸的最佳工艺参数为挤出频率30 Hz、滚圆频率50 Hz、滚圆时间4 min。采用流化床底喷上药包衣法制备的绞股蓝皂苷微丸体外释放度为95.98%,符合药典规定;脆碎度为0.90%;圆整度为0.80,表面较为光滑;堆密度较好、大小均匀;休止角为38.70°,流动性较好。电镜、红外、X-衍射等表征结果表明微丸圆整,绞股蓝皂苷分布均匀。利用挤出滚圆-流化床包衣法制备绞股蓝皂苷微丸,该工艺简便易行,可工业化生产。     

优质绞股蓝及其产品研制

以日本引种的201缢股蓝为原料,研制出含有丰富绞股蓝皂甙和氨基酸的绞股蓝饮料、绞股蓝口服液、绞股蓝饼干等保健产品。通过201绞股蓝在生产绞股蓝制品时的工艺特点以及我国一些省份的绞股蓝品种与201绞股蓝在总皂甙、总氨基酸含量上对比,证明此种绞股蓝为值得推广与栽培的优良品,是生产绞股蓝保健品的优质原料。     

高效液相色谱法测定保健食品中绞股蓝皂苷XLIX的含量

目的建立高效液相色谱法检测保健食品中绞股蓝皂苷XLIX含量的检测方法。方法样品经甲醇提取,采用Ultimate XB-C18柱(4.6 mm×100 mm, 3μm)进行色谱分离,以乙腈-水作为流动相进行梯度洗脱,流速为0.5 mL/min,检测波长为203 nm,柱温40℃。结果绞股蓝皂苷XLIX在0.001～1 mg/mL范围内线性关系良好(线性方程:Y=53.06X+86.3,r=0.999);绞股蓝皂苷XLIX在0.1、0.2、1.0g/kg~3个添加水平上进行加标回收实验,平均回收率为86.1%～104.9%,相对标准偏差为1.6%～6.0%,方法检出限为5mg/100g或5 mg/100 mL,定量限为10 mg/100 g或10 mg/100 mL。结论本方法前处理过程简单、分析时间短、准确度高,适用于保健食品中绞股蓝皂苷XLIX的检测。     

Reliable identification of grapevine rootstock varieties using RAPD PCR on woody samples

Since grapevine ( Vitis spp .) rootstock material is being traded increasingly as disbudded woody material a lack of distinctive morphological features on such material necessitates an alternative and reliable means of identification. Methods described here were developed for rapid and efficient extraction of DNA from woody samples rich in phenolic compounds and polysaccharides, and for subsequent identification of varieties by RAPD PCR. Using these methods, and with the application of only one selected RAPD primer, we were able to differentiate sixteen rootstock varieties, including the seven varieties most commonly used in Germany. Problems commonly encountered with reproducibility of RAPD patterns were avoided by choosing primers with a dinucleotide sequence and a high G/C content that allowed a rather high annealing temperature of 45°C. Methods described here should also be useful for other horticultural crops, especially those with woody tissues rich in phenolic compounds and polysaccharides.     

An internet compendium of analytical methods and spectroscopic information for monomers and additives used in food packaging plastics

An internet website (http://cpf.jrc.it/smt/) has been produced as a means of dissemination of methods of analysis and supporting spectroscopic information on monomers and additives used for food contact materials (principally packaging). The site which is aimed primarily at assisting food control laboratories in the European Union contains analytical information on monomers, starting substances and additives used in the manufacture of plastics materials. A searchable index is provided giving PM and CAS numbers for each of 255 substances. For each substance a data sheet gives regulatory information, chemical structures, physico-chemical information and background information on the use of the substance in particular plastics, and the food packaging applications. For monomers and starting substances (155 compounds) the infra-red and mass spectra are provided, and for additives (100 compounds); additionally proton NMR are available for about 50% of the entries. Where analytical methods have been developed for determining these substances as residual amounts in plastics or as trace amounts in food simulants these methods are also on the website. All information is provided in portable document file (PDF) format which means that high quality copies can be readily printed, using freely available Adobe Acrobat Reader software. The website will in future be maintained and up-dated by the European Commission's Joint Research Centre (JRC) as new substances are authorized for use by the European Commission (DG-ENTR formerly DGIII). Where analytical laboratories (food control or other) require reference substances these can be obtained free-ofcharge from a reference collection housed at the JRC and maintained in conjunction with this website compendium.     

Aroma characterization of various apricot varieties using headspace–solid phase microextraction combined with gas chromatography–mass spectrometry and gas chromatography–olfactometry

The characterization of the aromatic profile of several apricot cultivars with molecular tracers in order to obtain objective data concerning the aromatic quality of this fruit was undertaken using headspace–solid phase microextraction (HS–SPME). Six apricot cultivars were selected according to their organoleptic characteristics: Iranien, Orangered, Goldrich, Hargrand, Rouge du Roussillon and A4025. The aromatic intensity of these varieties measured by HS–SPME–Olfactometry were defined and classified according to the presence and the intensity of grassy, fruity and apricot like notes. In the six varieties, 23 common volatile compounds were identified by HS–SPME–GC–MS. Finally, 10 compounds, ethyl acetate, hexyl acetate, limonene, β-cyclocitral, γ-decalactone, 6-methyl-5-hepten-2-one, linalool, β-ionone, menthone and (E)-hexen-2-al were recognized by HS–SPME–GC–O as responsible of the aromatic notes involved in apricot aroma and considered as molecular tracers of apricot aromatic quality which could be utilized to discriminate apricot varieties.     

Tests of potential functional barriers for laminated multilayer food packages. Part I: Low molecular weight permeants

The advent of the functional barrier concept in food packaging has brought with it a requirement for fast tests of permeation through potential barrier materials. In such tests it would be convenient for both foodstuffs and materials below the functional barrier (sub-barrier materials) to be represented by standard simulants. By means of inverse gas chromatography, liquid paraffin spiked with appropriate permeants was considered as a potential simulant of sub-barrier materials based on polypropylene (PP) or similar polyolefins. Experiments were performed to characterize the kinetics of the permeation of low molecular weight model permeants (octene, toluene and isopropanol) from liquid paraffin, through a surrogate potential functional barrier (25 μm-thick oriented PP) into the food simulants olive oil and 3% (w/v) acetic acid. These permeation results were interpreted in terms of three permeation kinetic models regarding the solubility of a particular model permeant in the post-barrier medium (i.e. the food simulant). The results obtained justify the development and evaluation of liquid sub-barrier simulants that would allow flexible yet rigorous testing of new laminated multilayer packaging materials.