不同植物叶绿体中电子转移和能量传递超快过程

采用相同的分离技术,从三七、水葫芦和菠菜植物叶子中提取叶绿体.利用吸收光谱和低温荧光光谱及皮秒荧光单光子计数技术对这3种叶绿体的光谱性质和光系统Ⅱ荧光寿命进行了研究.这3种叶绿体吸收光谱相似,暗示着不同的植物都能高效吸收不同波长的光子.采用三指数动力学模型对测定的光系统Ⅱ荧光衰减曲线拟合,水葫芦植物叶绿体光系统Ⅱ荧光衰减寿命分别是:138,521和1494ps;菠菜体系叶绿体荧光寿命分别是:197,465和1459 ps;三七叶绿体体系荧光寿命分别是:30,274和805ps;并且归属了荧光组分,慢速度荧光衰减由叶绿素堆积造成,中等速度荧光衰减源于PSⅡ反应中心重新结合电荷组分,快速度荧光衰减归属于PSⅡ反应中心组分.定义并且基于20ps模型计算了三七、水葫芦和菠菜叶绿体光系统Ⅱ反应中心激发能转能效率,分别是60%,87%和91%.实验结果支持20 ps时间常数模型.三七叶绿体光系统Ⅱ反应中心低转能效率,2个光系统之间激发能分配平衡状态差的结果,以及它的光系统Ⅰ激发能红移的现象都与该植物生长速度慢的现象相吻合,显示植物生长速度特性可体现在光合作用原初过程中,表明植物生长速度与它的荧光性质及荧光寿命相关性,生长慢的植物对吸收的光子能量利用效率较低,而生长快的植物,转能效率则较高.     

水葫芦、三七和菠菜光系统Ⅱ颗粒的分离及超快光谱 的研究

分离了三七、水葫芦和菠菜植物光系统Ⅱ富集的颗粒,并且采用吸收光谱和低温荧光光谱及皮秒荧光单光子计数技术进行了研究。结果显示,它们的吸收光谱图具有相似性。采用三指数动力学模型对三种光系统Ⅱ颗粒实验测定的光系统Ⅱ荧光衰减曲线拟合。水葫芦植物PSⅡ颗粒光系统三个组分荧光寿命分别是:157ps,415ps和1661ps;菠菜体系的相应荧光寿命分别是:198ps,677ps和1244ps;三七体系的相应荧光寿命分别是:14ps,272ps和184ps。慢速度荧光衰减由叶绿素堆积造成的,中等速度荧光衰减源于PSⅡ反应中心重新结合电荷组分,快速度荧光衰减归属于PSⅡ反应中心组分。基于20ps模型计算出三七植物PSⅡ颗粒的光系统Ⅱ反应中心转能效率为41%,水葫芦为89%,菠菜为91%,数值结果表明三七植物生长慢的特性可以表现在光合作用原初过程中,大量的被值物吸收的光子产生的激发态能量以荧光发射和非辐射形式消耗了,而没有被有效地利用,进一步验证了我们提出的观点：植物生长速度与它们的荧光性质和荧光寿命有相关性,生长慢的植物对激发态能利用效率则较低。     

不同产地三七的~1H NMR指纹图谱对比分析

通过提取方法和检测方法的探讨,选用体积分数90%的甲醇水溶液,对三七主根样品进行超声提取,并用noesypr1d脉冲序列预饱和压制水峰实验,获取了8种不同产地三七的1H NM R指纹图谱,对三七主要成分特征信号进行了初步指认。通过图谱分析看出,不同产地三七的三萜皂苷含量和各种三萜皂苷组成比例不同,黄酮、糖类含量比重也不一样。由于1H NM R具有提供信息丰富、简便、快速和重现性好的特点,可以作为中药指纹图谱研究的方法。     

新型吸附树脂S—038对绞股蓝皂甙的吸附性能及其在绞股蓝皂甙和三七皂甙提取、纯化中的应用

绞股蓝皂甙、三七皂甙等达玛烷型皂甙可用作保健食品的添加剂和治疗药物。为发展其提取、纯化新技术,我们研究了S—038树脂对绞股蓝皂甙的静态吸附和动态吸附性能,发现该树脂具有较大的静态、动态吸附量。分别用该树脂从绞股蓝水渗滤液中提取、分离绞股蓝皂甙和从三七水浸提液中富集、纯化三七皂甙,结果提示该树脂适于这些皂甙的生产。     

多孔石墨化碳柱分析强极性化合物三七素

建立了强极性的非蛋白氨基酸三七素在石墨化碳色谱柱上的反相高效液相色谱分析方法。以Thermo Hypercarb石墨化碳柱(100 mm×4.6 mm,5 μm)为分离柱,以0.05 mol/L NaH2PO4(H3PO4调pH 2.2)为流动相,流速为1 mL/min,于柱温80 ℃、检测波长220 nm条件下分离检测。对三七素在石墨化碳柱上的保留及其影响因素的研究结果表明:三七素在石墨化碳柱上的保留机理仍主要是疏水相互作用。将建立的色谱条件用于三七药材中三七素的测定,进样量在0.22～4.4 μg范围内线性关系良好(r=0.9999),平均加标回收率为99.5%(n=9),相对标准偏差不高于2.2%,分析速度快(三七素的保留时间不到3 min)。     

三七水煎液中还原型谷胱甘肽及腺嘌呤的分离与鉴定

三七经水煎提取,阴、阳离子交换树脂分离以及RP-HPLC分离获得多种含肽类化合物组分,采用游离及酸水解后氨基酸组成及含量对照分析的方法预示了提取液中肽类化合物的存在,经氨基酸组成分析、聚丙烯酰胺薄膜双向层析、羧肽酶C-末端序列分析、质谱测定等方法分析,将YN-3H12组分鉴定为还原型谷胱甘肽,YN-3H11组分鉴定为腺嘌呤,本研究方法可为植物中水溶性肽类化合物等活性成分的研究提供参考。     

反相高效液相色谱法同时测定三七药材中4种皂苷的含量

建立了以0．02％磷酸-乙腈为流动相，梯度洗脱反相高效液相色谱同时测定中药材三七中三七皂苷R1、人参皂苷Rg1、Rb1和Rd 4种皂苷的新方法。R1、Rg1、Rb1和Rd 4种皂苷的加样回收率分别为89．54％、90．08％、82．82％与84．46％；线性范围分别为0．244-6．110、0．820-20．510、0．396-9．890与0．260-6．500μg。测定了不同规格、部位和来源的三七药材里的4种皂苷R1、Rg1、Rb1和Rd。方法准确可靠，结果稳定，重现性好，可用于三七及其制剂的质控。     

稀土元素超积累植物研究进展

概述了稀土元素的环境生态学特性及其超积累植物，并对稀土元素超积累植物研究的重要科学意义进行了介绍。在此基础上，详细综述了近年来稀土元素超积累植物的研究成果，包括其种属、空间地域分布，其体内稀土元素的分布、分异特征及影响因素，其吸收富集稀土元素的生理及生化机制，并展望了稀土元素超积累植物及其修复应用研究的发展前景。     

三七粗提液中皂甙与多糖泡沫分离的研究

测定了三七皂甙溶液的表面张力 ,考察了溶液 p H值、氮气流速、进料浓度等因素对泡沫分离的影响。实验结果表明 ,在 p H 7.0 0 ,进料浓度 2 .43× 1 0 2 μg· m L- 1(以人参皂甙 Rg1作参照 ,以下皂甙浓度均与此类同 ) ,三七皂甙的表面张力为 5 .93× 1 0 - 2 N· m- 1;氮气流速 1 5 m L· min- 1及进料体积 8.0 m L时 ,对三七粗提液进行泡沫分离 ,泡沫相三七皂甙收得率为 73.6%,液相三七多糖收得率为 87.5 %。本法具有简便、有效、无污染的优点     

砷对植物生长的影响及植物耐砷机理研究进展

砷污染问题是全球面临的重大环境污染问题之一。综述了不同浓度砷及不同形态砷对植物生长的影响，耐砷植物及植物耐砷机理，指出了今后研究的主攻方向。     

High-performance liquid chromatographic assay for the active saponins from Panax notoginseng in rat tissues

A reversed-phase liquid chromatographic method was used to determine the ginsenosides Rg1, Rb1 and Rd of Panax notoginseng in rat tissues (kidney, liver, heart, spleen and lung) after the administration of total saponins of P. notoginseng. The tissue samples were treated with solid-phase extraction prior to HPLC. The calibration curves for the three saponins were linear in the given concentration ranges. The intra-day and inter-day assay coefficients in tissues were between 76 and 120% respectively. The recoveries of all the tissues were higher than 70%. This method was applied to evaluate the distribution of the three major saponins of P. notoginseng in rat tissues.     

Hydrophilic interaction liquid chromatography with tandem mass spectrometry for the determination of underivatized dencichine (beta-N-oxalyl-L-alpha,beta-diaminopropionic acid) in Panax medicinal plant species

Dencichine (beta-N-oxalyl-L-alpha,beta-diaminopropionic acid) is a haemostatic agent present in important Chinese medicinal herbs such as Panax notoginseng, as well as other Panax species. It is also a reported neurotoxic agent found in Lathyrus sativus (grass pea seed). A selective analytical method incorporating hydrophilic interaction chromatography with positive electrospray ionization tandem mass spectrometry (HILIC/ESI-MS/MS), for the analysis of dencichine in Panax plant species, was developed. Using multiple reaction monitoring (MRM) mode, underivatized dencichine, a small and highly polar compound, was selectively detected and quantified. The contents of dencichine in raw and steamed Panax notoginseng roots, 11 pairs of raw and steamed P. notoginseng herbal products, Panax ginseng roots, and Panax quinquefolium roots, were analyzed and compared. Optimal sensitivity of 0.3 ppm (detection limit) and 1.5 ppm (quantification limit) was achieved. The method was rapid (< or =5 min), with the HILIC peak eluting at about 1 min. Steamed P. notoginseng samples were found to contain less dencichine than the corresponding raw samples, and there were also differences among the three Panax species; raw P. ginseng and P. quinquefolium contained less dencichine than the raw P. notoginseng species. This rapid and specific method may be applied to the quantification of dencichine in complex medicinal plants and their products.     

HPLC determination of four active saponins from Panax notoginseng in rat serum and its application to pharmacokinetic studies

Four main active saponins (ginsenosides Rg1, Rb1, Rd and notoginsenoside R1) in Panax notoginseng in rat serum after oral and intravenous administration of total saponins of P. notoginseng (PNS) to rats were determined using a simple and sensitive high-performance chromatographic method. The serum samples were pretreated with solid-phase extraction before analysis. The calibration curves for the four saponins were linear in the given concentration ranges. The intra-day and inter-day assay coefficients in serum were less than 10.0% and the recoveries of the method were higher than 80.0% in the high, middle and low concentrations. This method was applied to study the pharmacokinetics following oral and intravenous administration of PNS.     

Comparative study on chemical components and anti‐inflammatory effects of Panax notoginseng flower extracted by water and methanol

Methanol and water are commonly used solvents for chemical analysis and traditional decoction, respectively. In the present study, a high‐performance liquid chromatography with ultraviolet detection method was developed to quantify 11 saponins in Panax notoginseng flower extracted by aqueous solution and methanol, and chemical components and anti‐inflammatory effects of these two extracts were compared. The separation of 11 saponins, including notoginsenoside Fc and ginsenoside Rc, was well achieved on a Zorbax SB C18 column. This developed method provides an adequate linearity (r ² > 0.999), repeatability (RSD < 4.26%), inter‐ and intraday variations (RSD < 3.20%) with recovery (94.7–104.1%) of 11 saponins concerned. Our data indicated that ginsenoside biotransformation in PNF was found, when water was used as the extraction solvent, but not methanol. Specifically, the major components of Panax notoginseng flower, ginsenosides Rb1, Rc, Rb2, Rb3, and Rd, can be near completely transformed to the minor components, gypenoside XVII, notoginsenoside Fe, ginsenoside Rd2, notoginsenoside Fd, and ginsenoside F2, respectively. Total protein isolated from Panax notoginseng flower is responsible for this ginsenoside biotransformation. Additionally, methanol extract exerted the stronger anti‐inflammatory effects than water extract in lipopolysaccharide‐induced RAW264.7 cells. This difference in anti‐inflammatory action might be attributed to their chemical difference of saponins.     

Simultaneous determination of 11 saponins in Panax notoginseng using HPLC-ELSD and pressurized liquid extraction

A new HPLC coupled with evaporative light scattering detection (ELSD) method was developed for simultaneous determination of 11 major triterpene saponins, namely notoginsenoside R1 (1), ginsenosides Rg1 (2), Re (3), Rf(4), Rb1 (5), Rg2 (6), Rc (7), Rb2 (8), Rb3 (9), Rd (10), and Rg3 (11) in Panax notoginseng, a commonly used traditional Chinese medicine (TCM). Pressurized liquid extraction (PLE) was employed for sample preparation, and the analysis was achieved using a Zorbax ODS C18 column eluted with gradient water-ACN in 60 min. The drift tube temperature of ELSD was set at 60 degrees C, and nitrogen flowrate was at 1.4 L/min. The method provided good repeatability and sensitivity for quantification of 11 saponins with overall precision (including intra- and interday) and LOD of less than 2.9% (RSD) and 98 ng, respectively. The validated method was successfully applied to quantify 11 saponins in 28 samples of P. notoginseng collected in different places, which is helpful to control the quality of P. notoginseng and its related products.     

蛋白质组学技术鉴定与分析三七粉诱导海兔神经连索表达的差异蛋白质

RP-HPLC分离三七粉提取液,并鉴定含有Rb1、Rg1、Re、R1等皂甙成分。以蓝斑背肛海兔(Notarcusleachii cirrosus Stimpson,NLCS)为分析模型,三七粉提取液为诱导剂,选用蛋白质组技术研究NLCS神经连索诱导前后所表达的差异蛋白质。通过优化双向凝胶电泳分离NLCS神经连索全蛋白质组技术,获得496个蛋白质斑点。采用肽指纹图谱技术和数据库检索比对法,初步鉴定了NLCS受三七粉提取液诱导前后,其神经连索表达13个差异蛋白质,其中较高的匹配率蛋白质为肌动蛋白、3-羟酯酰辅酶A脱氢酶、ATP结合转运子和甲基转移酶12。选用LOC tree软件对13个差异蛋白质进行亚细胞定位,认为它们在保护神经系统中发挥重要的调节作用。     

Determination of Active Components in a Natural Herb with Near Infrared Spectroscopy Based on Artificial Neural Networks

The non-linear relationships between the contents of ginsenoside Rg1, Rb2, Rd and Panax notoginseng saponins(PNS) in Panax notoginseng root herb and the near infrared(NIR) diffuse reflectance spectra of the herb were established by means of artificial neural networks(ANNs). Four three-layered perception feed-for-ward networks were trained with an error back-propagation algorithm. The significant principal components of the NIR spectral data matrix were utilized as the input of the networks. The networks architecture and parameters were selected so as to offer less prediction errors. Relative prediction errors for Rg1, Rb1, Rd and PNS obtained with the optimum ANN models were 8.99%, 6.54%, 8.29%, and 5.17%, respectively, which were superior to those obtained with PLSR methods. It is verified that ANN is a suitable approach to model this complex non-linearity. The developed method is fast, non-destructive and accurate and it provides a new efficient approach for determining the active components in the complex system of natural herbs.     

改性松香键合二氧化硅高效液相色谱固定相的制备及其对三七总皂苷的分离

三七中发挥药效的主要成分为三七皂苷R1、人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1和人参皂苷Rd,用于贫血、冠心病、高血压、脑卒中后遗症等疾病的治疗,但其化学成分多且难分离.将氢化松香丙烯酸羟乙酯(HRHA)通过巯基-烯点击化学反应键合到烷基化硅胶表面,制备出一种新型的改性松香键合二氧化硅高效液相色谱固定相(SiO2...     

基质辅助激光解吸电离飞行时间质谱技术分析三七和海兔神经连索内的皂甙组分

基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱技术能高效解吸三七提取液(Panaxnotoginseng Extraction, PNE)中的混合皂甙分子为皂甙离子,并供质量分析器检测与分析.选用MALDI-TOF 质谱技术直接分析色谱纯皂甙样品的纯度,其检测灵敏度优于反相高效液相色谱法(RP-HPLC).优化提取中药三七(Panax notoginseng, PN)的混合皂甙, 选用MALDI-TOF质谱技术直接分析PNE中的皂甙种类和相对含量,发现PNE至少含有20种不同分子结构的皂甙组分,其中人参皂甙(Ginsenoside) Rg1和三七皂甙(Notoginsenoside)R1含量相对较高.选用薄板层析法(TLC)制备PNE中的R1皂甙.MALDI-TOF质谱技术研究蓝斑背肛海兔(Notarcus leachiicirrosus Stimpson, NLCS)神经连索内的超微量R1的组成与分布.建立PNE皂甙的指纹图谱,并适合于评价中药三七的品质和分析体内超微量皂甙的代谢过程与机理.     

Identification of ginsenoside metabolites in plasma related to different bioactivities of Panax notoginseng and Panax ginseng

Although the chemical components of Panax notoginseng (PN) and Panax ginseng (PG) are similar, their bioactivities are different. In this study, the differential bioactivities of PN and PG were used as the research object. First, the different metabolites in the plasma after oral administration of PN and PG were analyzed using a UPLC-Q/TOF-MS-based metabolomics approach. Afterward, the metabolite-target- pathway network of PN and PG was constructed, and thus the pathways related to different bioactivities were analyzed. As a result, 7 different metabolites were identified in PN group, and 10 different metabolites were identified in the PG group. In the PN group, the metabolite N1 was related to hemostasis, N1 and N3 were related to inhibiting the nerve center, antihypertensive, and abirritation. The metabolites N1, N3, N4, N5, and N6 were related to liver protection. The results showed that the metabolites G1, G2, G3, G5, and G6 in PG group were related to heart protection, and G1, G2, G6, and G9 were related to increased blood pressure. There were 13 signaling pathways related to different biological activities of PN (8 pathways) and PG (5 pathways). These pathways further clarified the mechanism of action that caused the different bioactivities between PN and PG. In summary, metabolomics combined with network pharmacology could be helpful to clarify the material basis of different bioactivities between PN and PG, promoting the research on PN and PG.