毛细管电泳-质谱联用技术研究进展

毛细管电泳-质谱（CE-MS）联用技术兼具毛细管电泳高效分离能力与质谱高灵敏检测、高真度定性的优势,已成为物质分离分析研究的一种非常重要的工具。本文对近几年来 CE-MS 联用的关键技术及 CE-MS 在中药分析、环境检测等领域的一些应用进展进行综述,对其发展进行了展望。     

毛细管电泳质谱联用技术及其在代谢组学研究中的应用

毛细管电泳自20世纪80年代中后期迅速发展以来,是近几年分析化学领域中发展最为迅速的分离手段之一。毛细管电泳与质谱的联用使得分析范围更广,灵敏度更高,因此被广泛应用于生物样品的分析中。介绍了毛细管电泳质谱联用常用的接口技术,质量分析器及其在生物样品中的分离模式,综述了近年来毛细管电泳质谱联用技术在代谢组学中的应用。     

铅在茶树体内的分布及化学形态特征

采用水培试验,结合亚细胞组分分离和化学形态提取技术,研究了茶树品种龙井43和迎霜不同器官的Pb含量,及其在根系亚细胞中的分布和化学结合形态.结果表明： 在Pb胁迫下,两个品种茶树根系的形态特征不同;龙井43新叶中Pb含量随营养液Pb浓度的提高显著增加,而迎霜的变化不显著.两个品种茶树根系亚细胞中Pb的含量分布及结合形态均存在差异,低浓度Pb处理下, 龙井43根亚细胞组分（除可溶物质外）中Pb含量均低于迎霜;而高浓度Pb处理下,龙井43根亚细胞组分（除细胞壁外）中Pb含量均高于迎霜;龙井43根中醋酸(HAc)提取态的Pb含量比例最高,其他化学形态的高低顺序为NaCl提取态>HCl提取态/H₂O提取态>乙醇提取态;而迎霜根中NaCl提取态Pb成分比例最高,其他化学形态的高低顺序为HAc提取态>HCl提取态/H₂O提取态>乙醇提取态.迎霜对Pb毒害的耐受能力强于龙井43.     

多维液相色谱分离技术在复杂蛋白质组学样品鉴定中的应用

目的：随着蛋白组学技术的发展,液相色谱-串联质谱的联用技术（液质联用）逐渐成为蛋白组学的主流技术。方法：通过结合各种不同原理的色谱分离类型,多维液相色谱分离技术能够极大的提高分离系统的峰容量,达到有效分离复杂程度很高的蛋白质组学样品的目的。结果：最广泛使用的多维液相色谱分离系统是离子交换色谱（IEX）和反相色谱（RP）的二维结合,近年来又发展出了分离能力更强的三维液相色谱分离系统,并且已经在蛋白质组学研究中得到了应用。结论：本文综述了多种多维液相色谱分离方法,在这些方法中,不同的分离原理的色谱类型被用于肽段或蛋白混合物的预分离中,有效促进了样品的充分分离,极大地提高了复杂样品的蛋白组学鉴定能力。     

蛋白质组学研究中的分离分析技术及其研究进展

在后基因组时代,蛋白质组学成为新的研究热点。蛋白质组学的研究目标是为复杂蛋白质样品建立一个高通量、大规模、自动化的分离分析技术平台,从而实现准确、快速地筛选功能蛋白质。蛋白质的分离分析在蛋白组学研究中起着非常重要的作用。本文主要综述在蛋白质组学研究中二维凝胶电泳、毛细管电泳及其与质谱联用、多维液相分离技术及其与质谱联用和蛋白质芯片等高效分离分析技术的应用研究进展。     

蛋白质组学研究中的对角线电泳

经典的蛋白质组学研究方法包括IEF/SDS-PAGE双向电泳和质谱技术的联用,但由于IEF的一些不足,限制了其应用范围。对角线电泳是蛋白质组学研究中的一项特殊分离技术,由于其原理与IEF/SDS-PAGE不同,正逐渐成为蛋白质组学中电泳分离技术的重要补充,特别是在膜蛋白和蛋白质相互关系的研究中将起到重要作用。本文综述了对角线双向电泳技术的特点、发展和在蛋白质组学研究中的最新进展,比较了双向电泳和对角线电泳的优缺点,展望了对角线电泳在蛋白质组学研究中的应用前景。     

生物样品中硒的形态分析方法研究进展

对近年来关于生物样品中微量元素硒的形态分析方法的研究成果进行了综述,总结了高硒生物样品(10 mg Se/kg)和低硒生物样品(1 mg Se/kg)的硒形态组分提取方法,以及利用液相色谱-原子荧光光谱(LC-AFS)、高效液相色谱与电感耦合等离子质谱(HPLC-ICP-MS)等仪器联用的方法分离、检测硒的形态组分。此外,对利用同步辐射X射线微区分析方法(STXM)原位微区分析微生物样品的硒形态及其分布进行了综述与展望。     

蛋白质组学中的分离检测技术

10多年来,随着基因组学研究取得的巨大成就,蛋白质组学的研究也得到了突飞猛进的发展,并产生了许多先进的分离检测技术,包括与电泳相关的和非电泳的技术。本就蛋白质组学中的分离检测技术,如双向电泳、差异凝胶电泳、毛细管电泳、液相色谱质谱联用、蛋白质芯片等作一综述。     

蛋白质组研究中分离新技术与新方法

对于蛋白质组的研究离不开分析技术的支撑。由于样品及其基质的复杂性,为了实现蛋白质的高通量、高灵敏度、快速分析鉴定,必须发展与之匹配的新技术与新方法。多维高效液相色谱/毛细管电泳技术,部分弥补了传统2D PAGE的不足,近年来,在蛋白质分离鉴定方面取得了最令人瞩目的成绩。本文分别从多维液相色谱分离技术、多维毛细管电泳蛋白质分离平台、微柱液相-毛细管电泳联用技术、极端pH蛋白质的分离分析和蛋白质的在线富集技术等方面对蛋白质组学研究中在新技术与新方法方面近期取得的成果加以系统阐述。     

土壤中砷的形态转化及其分析方法

在分析土壤中砷的来源及存在形态的基础上,简要探讨了不同形态砷之间相互转化的过程.对土壤砷形态的提取和检测方法进行了较系统的比较,认为高效液相色谱-氢化物发生-原子荧光光谱（HPLC-HG-AFS）联用技术具有灵敏度高、检测限低、选择性强、运行成本低、不同形态砷之间相互转化较少的优点,可以作为砷形态检测的首选方法.结合相关研究结果,对土壤中砷形态转化及相关研究的发展方向进行了讨论.     

Partitioning of trace metals in sediments: Relationships with bioavailability

As a result of complex physical, chemical and biological processes, a major fraction of the trace metals introduced into the aquatic environment is found associated with the bottom sediments, distributed among a variety of physico-chemical forms. As these different metal forms will generally exhibit different chemical reactivities, the measurement of the total concentration of a particular metal provides little indication of potential interactions with the abiotic or biotic components present in the environment. In principle, the partitioning of sediment-bound metals could be determined both by thermodynamic calculations (provided equilibrium conditions prevail) and by experimental techniques. The modelling of sediment-bound metals is far less advanced than is that of dissolved species, primarily because the thermodynamic data needed for handling sediment-interstitial water systems are not yet available. The partitioning of a metal among various fractions obtained by experimental techniques (e.g., sequential extraction procedures) is necessarily operationally defined. These methods have, however, provided significant insight into the physico-chemical factors influencing the bioavailability of particulate trace metals; some of these factors are discussed.     

Synchrotron radiation for direct analysis of metalloproteins on electrophoresis gels

Metalloproteomics requires analytical techniques able to assess and quantify the inorganic species in metalloproteins. The most widely used methods are hyphenated techniques, based on the coupling of a high resolution chromatographic method with a high sensitivity method for metal analysis in solution. An alternative approach is the use of methods for solid sample analysis, combining metalloprotein separation by gel electrophoresis and direct analysis of the gels. Direct methods are based on beam analysis, such as lasers, ion beams or synchrotron radiation beams. The aim of this review article is to present the main features of synchrotron radiation based methods and their applications for metalloprotein analysis directly on electrophoresis gels. Synchrotron radiation X-ray fluorescence has been successfully employed for sensitive metal identification, and X-ray absorption spectroscopy for metal local structure speciation in proteins. Synchrotron based methods will be compared to ion beam and mass spectrometry for direct analysis of metalloproteins in electrophoresis gels.     

On available forms of chemical compounds in soils

Different methods are used to study available forms of chemical compounds in soils. The sequential extraction method is described in detail. Also, a comparative overview is given of the most frequently extracted fractions of chemical elements (water-soluble, exchangeable, specifically sorbed, carbonate, bonded to organics, Fe and Mn oxides/hydroxides, and aluminosilicates), as well as of relevant extractants. Advantages and disadvantages of various extractants are discussed, and ways to improve their selectivity are proposed. It is also shown that the content of available forms of chemical compounds in soil can be used for estimating the extent of industrial soil pollution.     

微生物分子生态学技术及其在环境污染研究中的应用

较为系统地概述了核酸探针检测技术、利用引物的PCR技术、DNA序列分析技术和电泳分离及显示技术在国内外的研究进展,并探讨了这些技术在环境污染研究中的应用及其方向。结果表明,这些被认为是重要的微生物分子生态学技术,在探索微生物与污染环境之间的相互关系中发挥了重要作用。促进了污染环境的微生物遗传适应进化机制的研究,污染物的微生物降解有关基因的定位及微生物工程菌的构建等方面的工作,从而推进了污染环境微生物修复的分子生态学的发展。     

Precision genetic modifications: a new era in molecular biology and crop improvement

Recently, the use of programmable DNA-binding proteins such as ZFP/ZFNs, TALE/TALENs and CRISPR/Cas has produced unprecedented advances in gene targeting and genome editing in prokaryotes and eukaryotes. These advances allow researchers to specifically alter genes, reprogram epigenetic marks, generate site-specific deletions and potentially cure diseases. Unlike previous methods, these precision genetic modification techniques (PGMs) are specific, efficient, easy to use and economical. Here we discuss the capabilities and pitfalls of PGMs and highlight the recent, exciting applications of PGMs in molecular biology and crop genetic engineering. Further improvement of the efficiency and precision of PGM techniques will enable researchers to precisely alter gene expression and biological/chemical pathways, probe gene function, modify epigenetic marks and improve crops by increasing yield, quality and tolerance to limiting biotic and abiotic stress conditions.     

Chromatographic and electrophoretic approaches in ink analysis

Inks are manufactured from a wide variety of substances that exhibit very different chemical behaviors. Inks designed for use in different writing instruments or printing methods have quite dissimilar components. Since the 1950s chromatographic and electrophoretic methods have played important roles in the analysis of inks, where compositional information may have bearing on the investigation of counterfeiting, fraud, forgery, and other crimes. Techniques such as paper chromatography and electrophoresis, thin-layer chromatography, high-performance liquid chromatography, gas chromatography, gel electrophoresis, and the relatively new technique of capillary electrophoresis have all been explored as possible avenues for the separation of components of inks. This paper reviews the components of different types of inks and applications of the above separation methods are reviewed.     

Isolation, characterization, and amino-terminal amino acid sequence analysis of human neutrophil elastase from normal donors

Human neutrophil elastase from normal donors has been purified using an isolation procedure which included sequential sodium chloride extraction, Aprotinin-Sepharose affinity chromatography, CM-cellulose ion-exchange chromatography, and AcA44 gel filtration chromatography. The inclusion of this last purification step was crucial for separating inactive lower molecular weight species from the active forms of neutrophil elastase and resulted in a higher specific activity of the final preparation. Sodium dodecyl sulfate-polyacrylamide gradient gel electrophoresis of the reduced purified protein demonstrated three polypeptides of Mr 31,000, 28,000, and 27,500. Four polypeptides were resolved on acid gel electrophoresis; each of the four possessed amidolytic activity. Furthermore, peptide analysis of Staphylococcus aureus V8 protease digests indicated that these polypeptides are structurally related to each other and represent microheterogeneity of the purified protein. The apparent isoelectric points of these four forms as determined by two-dimensional electrophoresis range from 6.1 to 6.7. By utilizing microsequencing techniques, the first 40 residues of neutrophil elastase have been determined and compared with the reported sequence of elastase isolated from leukemic myeloid cells. In addition, a high degree of homology was found within the amino-terminal regions of neutrophil elastase and the serine proteinases porcine elastase, bovine chymotrypsin, human factor D, and the beta chain of plasmin.     

Separation and identification methods for metalloproteinase inhibitors

Metalloproteinase inhibitors are being explored for the treatment of a wide variety of human diseases including cancers, arthritis, cardiovascular disorders, human immunodeficiency virus infection, and central nervous system illnesses. This review provides an overview of various analytical sample preparation, separation, detection, and identification techniques employed for the quantitative and qualitative determination of these inhibitor compounds. Special emphasis is placed on biological sample preparation by automated solid-phase extraction, liquid–liquid extraction, and protein precipitation by centrifugation or filtration. Other sample preparation methodologies are also evaluated. Applications of high-performance liquid chromatography, gas chromatography, and capillary electrophoresis to the quantitative determination of metalloproteinase inhibitors are described. Examples of qualitative analysis of metalloproteinase inhibitors by hyphenated liquid chromatography with mass spectrometry and nuclear magnetic resonance are also presented. The advantages and limitations of these separation and identification methodologies as well as other less frequently employed techniques are assessed and discussed.     

Present and potential applications of mass spectrometry for bioprocess research and control.

For on-line monitoring of bioprocesses present applications are mainly restricted to gas analysis, but several techniques have been improved recently: membrane probes, the application of MS/MS techniques, methods of correlating available on-line data like gas reaction rates with bioprocess characteristics using stoichiometric models and other empirical correlations. New ionization and ion separation methods for biomolecules are developing dramatically. Most striking developments in this area are improved desorption techniques, electrospray, the renaissance of time-of-flight instruments and new challenges in ion trap techniques. Enormous progress is made in the analysis of peptides and other biopolymers. Combinations with new separation techniques like capillary electrophoresis and capillary HPLC show new horizons in biomolecule analysis.     

污染土壤重金属的生物有效性和移动性评价：四种方法比较

重金属在土壤中的积累可增加土壤对生态环境的危害,而这种危害与土壤中重金属的活性有关.本文以植物体重金属浓度和地表径流重金属浓度为依据,比较研究了总量法、化学形态分级法、有效成分提取法和淋洗法4种方法评价污染土壤中重金属的生物有效性和移动性的可行性.结果表明,不同方法的评价结果有较大的差异.由于不同土壤的重金属组成有很大的差异,总量法难以反映土壤重金属的生物有效性和移动性;化学形态分级法中的交换态重金属可较好地反映土壤重金属的生物有效性和移动性,有机质结合态和碳酸盐结合态的某些重金属与其生物有效性和移动性也有一定的联系,而氧化物结合态、残余态重金属与重金属的生物有效性和移动性无关;用淋洗方法溶出的重金属量可很好地反映地表径流中重金属的浓度,也可较好地反映重金属的生物有效性;5种化学提取剂提取有效态重金属的结果表明,稀盐(0.01 mol·L^-1 CaCl₂)和1 mol·L^_-1 NH₄OAc提取的土壤重金属量与植物中重金属的积累和地表径流中重金属浓度均显著相关,可较好地表征土壤中重金属的生物有效性和移动性,其中稀盐(0.01 mol·L^-1 CaCl₂)提取的重金属最适于评价重金属的可移动性.