毛细管区带电泳分离小麦种子醇溶蛋白的研究

利用毛细管区带电泳（ＣＺＥ）检测了在不同电泳条件下分离小麦种子醇溶蛋白的效果。结果显示,用碱性缓冲液（０．０６ｍｏｌ／Ｌ硼酸钠,ｐＨ９．０,并含有２０％乙腈和１％ＳＤＳ）及４７ｃｍ×５０μｍｉ．ｄ．毛细管柱,在１５ｋＶ电压和３０℃的条件下可获得较高的分辨度和重复性。     

基于非靶标代谢组学方法研究水稻种子的代谢物轮廓

建立了一种基于气相色谱-质谱联用的非靶标代谢组学方法,研究了不同品种水稻种子之间的代谢物差异。分别考察了3种提取方式及4种提取溶剂对水稻中代谢物提取效率的影响,发现涡旋提取方法及80%甲醇提取可获得较好的结果,该方法具有良好的稳定性和线性。将方法用于3种水稻种子(农大、稻花香、状元)的代谢轮廓分析,结果显示3种水稻的代谢特征不同。相对于农大种子,稻花香种子中脂肪酸(棕榈酸、油酸、亚油酸、硬脂酸)和糖类(葡萄糖、半乳糖、甘露醇)、天冬酰胺、柠檬酸的含量较高,此外,状元种子中乳酸含量也较高;稻花香种子中琥珀酸、果糖,状元种子中的苏氨酸含量低于农大种子。但相对于状元种子,稻花香中乳酸、琥珀酸、硬脂酸、果糖等物质含量较低。该方法可应用于不同品种、不同基因型或不同产地水稻种子的代谢表型分析,为改善水稻产量及质量等研究提供信息。     

毛细管电泳研究和应用的最新进展

以作者所在实验室近期工作为主线，结合第五届国际毛细管电泳学术报告会（１９９３．１．２５－２８，Ｏｒｌａｎｄｏ，ＵＳＡ）的有关背景情况，介绍了近年来高性能毛细管电泳在应用、技术和理论方面研究工作的进展。     

一种新型毛细管电泳—电感耦合等离子体光谱法接口装置

毛细管电泳是一种高效、简便的分离方法，已被用于生物、环境及临床等试样的分离及分析[1]．检测技术在毛细管电泳中占有重要的地位，目前，在柱紫外可见及荧光检测是两种广为接受的检测方法，但其检测灵敏度仅为10－5～10－6mol／L[2]．电感耦合等离子体光谱（ICP－AES／MS）是一种灵敏的元素选择性的分析方法，已被广泛地用于各种试样中元素分析．近年来，该方法作为色谱及毛细管电泳的检测器，被用于元素的形态分析[3]．在毛细管电泳（CE）与ICP光谱连用技术中，挑战性的工作是设计一种能把CE与ICP相连的接口．目前已有几种接口…     

分析化学

毛细管电色谱（CEC）集毛细管电泳的高柱效和高效液相色谱的高选择性为一体，在药物和生物分析领域有着巨大的：应用潜力。最近，天津大学药物科学与技术学院的万谦宏研究组在Anal．Chem，2007，79（13）：5082～5086发表的论文中，报道了制备毛细管电色谱柱的新方法。利用磁场力将键合十八烷基的磁性微球（Magnetic Microspheres，MM）固定于毛细管中，得到新颖的无塞CEC柱，应用于中性混合物的分离分析，结果可以和其它CEC填充柱相媲美。     

羟丙基-β-环糊精的取代度和取代位置对毛细管电泳手性药物分离的影响

研究手性化合物的分离分析方法是当今国际分析化学领域研究的热点．毛细管电泳（CE）手性分离具有高效、快速、低耗等特点，近年来已取得很大进展［‘j．环糊精（CD）及其衍生物是CE手性分离使用最多的手性选择剂，其手性分离机理是基于手性化合物（客体）的2个对映体与环糊精分子（主体）形成的包合物的稳定常数不同，而这种差别主要取决于主客体分子之间的匹配性．因此，CD及其衍生物的分子结构，包括CD衍生物的取代度和取代位，置，对手性识别有很大影响．目前，有关CD及其衍生物作为手性选择剂的CE手性分离报道较多“‘，研究的主…     

基体辅助激光解吸电离飞行时间质谱法测定钙调素的分子量

近几年来，由于各种“软”电离技术的发展和应用「‘，’‘，为利用质谱分析生物大分子提供了一条十分有效的途径［‘j，其中基体辅助激光解吸电离飞行时间质谱法（MALDI－TOF－MS）是最有前途的技术之一D‘．钙调素（CaM）是动植物中普遍存在的多功能的胞内钙受体蛋白，参与调控许多生理反应［”．最近几年也在细胞外区域发现了钙调素及其功能［”．钙调素分子量的准确、快速测定是钙调素性质、结构和功能研究的首要问题．目前CaM分子量的测定大多采用SDS－PAGE、凝胶过滤和沉淀平衡等方法，例如对牛脑CaM分子量的SDS－PAGE测…     

人的基因组及DNA序列分析──过去、现去及将来（上）

人的基因组研究已成为生命科学前沿领域中最热门的课题之一。ＤＮＡ序列分析是基因组研究的关键技术．本文对人的基因组分析及其对ＤＮＡ序列分析的要求进行了论述．对ＤＮＡ序列分析方法如板凝胶电泳自放射显影法、板凝胶电泳激光荧光法、毛细管电泳激光荧光法、阵列毛细管凝胶电泳激光荧光法。超薄层板在胶电泳激光荧光法作了详细评论．并对正在开发的不用凝胶电泳分离的直接测序新技术和新方法，如质谱法、原子探针法（扫描隧道显微镜、原子力显微镜）、杂交法、流动单分子荧光检测法等进行了评论。     

固相萃取-气相色谱-质谱联用法测定人尿中11种蛋白同化激素

建立了固相萃取（SPE）-气相色谱-质谱联用（GC-MS）同时测定尿样中11种蛋白同化激素的分析方法,并将该方法用于关雄酮（MADE）阳性尿的分析。尿样前处理采用含500mg填料的C18 SPE柱净化浓缩,然后进行酶解,最后由液-液萃取获得待测样品。为了提高灵敏度,待测样品采用硅烷化衍生后进行GC-MS分析。在选择离子监测（SIM）模式下,采用甲睾酮（MT）为内标进行定量分析,方法检出限为0．1～2．0μg·L^-1;分析加标实际样品,回收率在70．5％～121％,相对标准偏差为1．1％～13．9％。     

毛细管区带电泳用于分离分析砷化合物的研究

采用毛细管区带电泳，以磷酸盐为缓冲溶液分离了5种砷的化合物：As（Ⅲ）、DMA、ANA、MMA和As（Ⅴ）．用紫外吸收法在191nm波长下，对实验条件进行了优化；使用扩展光程毛细管研究了实验方法的重现性；测定了各种砷化合物的峰面积与浓度的关系，在最大进样量时测定了检出限，并将该方法用于合成样品的分析．     

Up-scaling capillary zone electrophoresis separations of polydisperse anionic polyelectrolytes with preparative free-flow electrophoresis exemplified with a soil fulvic acid

A scale-up of analytical capillary zone electrophoresis (CZE) to preparative free-flow electrophoresis (FFE) is described. FFE allows fractionations based on charge densities in larger amounts than in CZE, enabling further off-line analysis of the fractions. Model compounds (carboxylic acids and polystyrene sulfonates) showed a similar behavior in FFE as in CZE. Diffusion and electrodynamic distortion effects are more pronounced in FFE than in CZE. A soil fulvic acid was analyzed by CZE and fractionated by FFE. A comparison of the FFE fractions with CZE measurements of the same sample using the effective mobility scale showed good agreement of the two methods.     

Separation of nicotine metabolites by capillary zone electrophoresis and capillary zone electrophoresis/mass spectrometry

The use of capillary zone electrophoresis (CZE) and capillary zone electrophoresis/mass spectrometry (CZE/MS) has been demonstrated, in principle, for the separation of nicotine and nicotine metabolites. The buffer system developed for separation and detection by CZE/UV was modified for use in CZE/MS analysis. Several of the metabolites are isobaric and tandem mass spectrometric (MS/MS) techniques have been used to differentiate such analytes.     

Free solution capillary electrophoresis of proteins using untreated fused-silica capillaries.

Numerous efforts have been made to separate proteins by capillary zone electrophoresis (CZE). The most common optimization techniques are changing the pH of the running buffer, coating the capillary surface with a hydrophilic polymer, or using additives in the sample solution. Surface coatings and solution additives can reduce the adsorption of the protein onto the capillary surface, but they diminish the separation efficiency and the resolution of CZE. This paper reports the successful separation of proteins in a untreated fused-silica capillary by raising the pH of the running buffer and washing between runs with 1.0 M sodium hydroxide. Under these conditions, model proteins and proteins in human serum have been determined by CZE. It is shown that the results from CZE are compatible with those of sodium dodecyl sulphate-polyacrylamide gel electrophoresis.     

寡糖的毛细管电泳分析

多种寡糖经α－萘胺衍生化后，用硼砂作为电泳介质，实现了高效毛细管电泳分离。比较了毛细管区带电泳和胶束毛细管电动色谱分离寡糖α－萘胺衍生物的电泳行为，对影响分离度的诸因素进行了考察，选择了最佳分离条件。     

Integrated electroosmotically-driven on-line sample purification system for nanoliter DNA sequencing by capillary electrophoresis

An integrated on-line system is developed for DNA sequencing at the nanoliter scale. The technique involves the use of a nanoreactor for small-volume cycle-sequencing reaction, capillary zone electrophoresis (CZE) for purification of the sequencing fragments, and capillary gel electrophoresis (CGE) for separation of the purified DNA fragments. The nanoreactor and CZE are integrated into one capillary, where a 100-nl dye-labeled terminator cycle-sequencing reaction is carried out followed by CZE to separate excess dye-labeled terminators from the sequencing fragments. On-line electrokinetic injection of the purified DNA fragments into the CGE system is accomplished at a small-volume tee connector by which the CZE capillary is interfaced to the CGE system. The utility of the system is demonstrated in sequencing nanoliter volumes of single-stranded DNA (M13mp18) and double-stranded DNA (pGEM). The use of voltage to drive both CZE and CGE makes it feasible for automation and future adaptation of the whole system to a microchip.     

Correlation of capillary zone electrophoresis with continuous free-flow zone electrophoresis: application to the analysis and purification of synthetic growth hormone releasing peptide

Two carrier-free electrophoretic separation methods, capillary zone electrophoresis (CZE) and continuous free-flow zone electrophoresis (FFZE), have been applied to both microanalysis at the nanogram level and preparative fractionation, with a throughput of 30 mg/h, of synthetic growth hormone releasing peptide (GHRP). A crude product of GHRP, a hexapeptide with the sequence His-D-Trp-Ala-Trp-D-Phe-Lys-NH2, synthesized by the solid phase methodology, was desalted and analyzed by CZE. Based on the results of analytical CZE the separation was converted into a preparative purification procedure by continuous FFZE, employing the same separation medium (0.5 mol/L acetic acid, pH 2.6). The purifity of peptide fractions obtained by FFZE was reevaluated by CZE. The combination of these two techniques proved to be a valuable tool for both peptide analysis and peptide purification. A close correlation of CZE and FFZE, resulting from the fact that both methods are based on the same separation principle (zone electrophoresis) and that both are performed in a free solution of the same composition, was confirmed. However, when transforming data from CZE to FFZE, the different electroosmotic flow, temperature and electric field intensity in the capillary and in the flow-through cell, respectively, have to be taken into account and corresponding corrections have to be made.     

Comprehensive on-line RPLC-CZE-MS of peptides

Peptide standards and tryptic digests of ribonuclease B are separated by comprehensive two-dimensional reversed phase liquid chromatography (RPLC) and capillary zone electrophoresis (CZE) and detected on-line by electrospray mass spectrometry. The RPLC column is coupled to the CZE column by a transverse flow gating interface. A new rugged microelectrospray needle is described that combines high ionization efficiency, low flow rates, and a sheath flow. The result is a system combining the separation capabilities of both RPLC and CZE with on-line mass spectrometric detection, all in about 15 min.     

Complete assignment of bilberry (Vaccinium myrtillus L.) anthocyanins separated by capillary zone electrophoresis

Capillary zone electrophoresis (CZE) mobilities of fifteen anthocyanins in bilberry extract were completely characterized. Four minor anthocyanins in bilberry extract (malvidin 3-O-alpha-L-arabinopyranoside (Mv 3-ara), peonidin 3-O-beta-D-galactopyranoside (Pn 3-gal), peonidin 3-O-alpha-L-arabinopyranoside (Pn 3-ara), and petunidin 3-O-alpha-L-arabinopyranoside (Pt 3-ara)) that remained unidentified in our previous CZE study were isolated from the bilberry extract, and the chemical structures were assigned by NMR and MS. Their CZE mobilities were then precisely examined together with those of other major anthocyanins in the extract. When the CZE mobilities of the fifteen anthocyanins assigned here were plotted against their molecular weight/numbers of free phenolic group, it was found that separation of anthocyanins by CZE is primarily determined by the type of conjugated sugar present, and secondly by the aglycon structure.     

Quantitative capillary zone electrophoresis of inorganic anions

Despite the availability of commercial capillary electrophoresis systems for over ten years, where quantitative analysis is required, capillary zone electrophoresis (CZE) has often failed to replace ion chromatography as the method of choice for a large number of analytes, not least inorganic anions. To investigate the reasons for this apparent failing, a review is presented of work that has been carried out to-date involving the quantitative application of CZE to the determination of inorganic anions in industrial and environmental samples. This review summarizes work both investigating and improving the quantitative aspects of the CZE of inorganic anions. A complete survey of how CZE has been applied to the determination of inorganic anions in real samples is given, including what, if any, analytical performance parameters were investigated and quoted, and if quality assurance data and validation methods were briefly considered, thoroughly investigated or simply ignored.     

Capillary zone electrophoresis and capillary electrophoresis-mass spectrometry for analyzing qualitative and quantitative variations in therapeutic albumin

The present study describes a reproducible and quantitative capillary zone electrophoresis (CZE) method, which leads to the separation of nine forms (native, oxidized and glycated) of human serum albumin (HSA). In an attempt to identify the different species separated by this CZE method, the capillary electrophoresis was coupled to mass spectrometry using a sheath liquid interface, an optimized capillary coating and a suitable CE running buffer. CE-MS analyses confirmed the heterogeneity of albumin preparation and revealed new truncated and modified forms such as Advanced Glycation End products (AGEs). Assignment of the CZE peaks was carried out using specific antibodies, carboxypeptidase A or sample reduction before or during the CE separation. Thus, five HSA forms were unambiguously identified. Using this CZE method several albumin batches produced by slightly different fractionation ways could be discriminated. Furthermore, analyses of HSA preparations marketed by five pharmaceutical industries revealed that two therapeutic albumins, including that marketed by LFB, contained the highest proportion of native form and lower levels of oxidized forms.