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1.
毛细管液相色谱柱具有节省样品、流动相,固定相消耗少,提高检测灵敏度等优点[1].近年来,随着蛋白组学的发展,毛细管离子交换液相色谱受到重视.目前蛋白组学中占有重要地位的多维液相色谱-质谱联用技术中,第一维液相色谱大多采用毛细管离子交换色谱柱[2].目前采用的颗粒填充毛细管柱柱压较高,易被蛋白质污染,且价格昂贵,使用寿命短.整体柱[3]具有大孔结构,柱背压低,对流传质使传质速率加快,制备过程简单,由于溶质与基质的接触时间短,降低了生物样品失活的可能性.与强阳离子交换固定相相比较,弱阳离子交换固定相与生物分子的相互作用比较温和,因此不易发生蛋白质变性现象.目前报道的弱阳离子交换整体柱通常采用两步法改性[4]:先用乙二胺处理,再用氯乙酸反应.我们在选取的具有良好刚性和渗透性的整体固定相上,利用亚胺乙二酸进行一步改性,简化了合成步骤,获得了具有羧酸基团的弱阳离子交换毛细管整体柱,并考察了其对于蛋白的分离能力. 相似文献
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研究了一些芳胺类化合物在1-萘甲膦酸改性氧化锆固定相上的色谱行为。分别考察了流动相中甲醇含量、缓冲液pH值和离子强度等对芳胺类化合物色谱保留的影响,并对这类化合物在该固定相上的保留机理进行了探讨。研究结果表明,芳胺类化合物在该固定相上表现出反相和阳离子交换的混合保留模式。以pH 10.1的Tris-甲醇(60/40,V/V)溶液为流动相,在1-萘甲膦酸改性氧化锆固定相上成功分离了间苯二胺、邻甲苯胺、N-甲苯胺、对硝基苯胺、邻硝基苯胺和α-甲萘胺6种芳胺类化合物。 相似文献
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采用拓扑指数(mQ)、定位基参数(SOX)及固定液极性值(CP)与脂肪醇在6种固定相上的气相色谱保留指数值(RI)进行了相关分析,发现RI与上述参数间存在良好的相关性,其关系可表示为:RI=158·384 57·7220Q-12·0491Q 5·720SOX 5·756CP R=0·9913继以留一法(Leave-one-out,LOO)进行交互检验,相关系数RCV=0·9909。说明所建定量结构-保留关系(QSRR)模型具有良好的稳定性和预测能力,较好地揭示了脂肪醇类物质在不同固定相上气相色谱保留指数的变化规律。 相似文献
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离子色谱是分离分析阳离子型化合物的重要手段之一。高效阳离子交换固定相的制备研究对离子色谱技术的发展具有重要的意义。该文以丙烯酸和顺丁烯二酸酐为单体,2-巯基乙基磺酸钠为巯基改性剂,提出了聚合物基质微球巯基改性自由基聚合修饰方法,用以制备新型双功能的阳离子交换固定相。该固定相以羧基和磺酸基为功能基,仅用简单的强酸淋洗液便可以实现常规阳离子的基线分离。利用色谱学模型,对金属离子和有机胺的保留行为进行了研究。采用梯度淋洗模式,可在24 min内实现10种阳离子的分离,表明固定相具有优异的色谱性能。“巯基-烯”修饰方法简单、高效。此外,通过调节巯基改性剂的比例能够实现对固定相交换能力的调控。 相似文献
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以新型环保多齿螯合剂——亚氨基二琥珀酸(IDS)为配体,在优化条件下,合成了IDS-Silica固定相。用电位滴定法测定了固定相上IDS的键合量。考察了IDS-Silica柱的色谱特性以及金属螯合特性。使用制备柱成功地分离了标准蛋白质混合物,该制备柱展现出了典型的阳离子交换特性。用电感耦合等离子体原子发射光谱法考察了金属离子在IDS-Silica固定相上的键合特性。结果表明,金属离子在IDS-Silica固定相上键合量的变化规律与它们同该固定相螯合的强弱顺序一致。通过比较金属Cu~(2+)在4种不同氨羧类配体硅胶柱上的键合量,发现IDS对金属离子具有强的螯合能力。IDS对金属离子的强螯合特性为其今后作为固定金属亲和色谱填料奠定了基础,为缓解亲和柱在使用过程中固定金属离子的流失提供了一种有效的解决方法。 相似文献
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单分散树脂基质的弱阳离子交换色谱固定相的制备及其在生物大分子分离中的应用研究 总被引:2,自引:0,他引:2
采用分散聚合法制备种子和“一步种子溶胀聚合法”制备了粒径为6~15 μm的单分散多孔氯甲基苯乙烯-二乙烯苯微球。该微球经化学改性后得到一种亲水性良好的新型高效弱阳离子交换色谱固定相。详细考察了该固定相的表面亲水性、对标准蛋白的分离性能和盐的种类对蛋白质保留行为的影响。考察结果表明该固定相是一种性能优异的弱阳离子交换色谱固定相。将其应用于鸡蛋清中溶菌酶的快速分离纯化,纯化后的溶菌酶纯度高于96%,比活高达71184 U/mg。 相似文献
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为使混合机理色谱(MMC)得到广泛地应用, 合成、表征和评价MMC固定相就成了首先要解决的问题. 依据离子交换色谱柱也具有疏水色谱(HIC)保留机理的特征, 选了4种弱阳离子交换(WCX)柱和一根二维[2D(WCX,HIC)]色谱柱, 研究了标准蛋白在这两类色谱柱上的保留行为. 这四种WCX色谱柱中的两种能在WCX和HIC两种分离模式下分离蛋白, 虽不如2D色谱柱效果好, 但有可能当成“准2D柱”来使用. 发现蛋白在这四种WCX柱上所显示的HIC分离特征各不相同, 且保留值随盐浓度变化呈现出的“U型”曲线也有大的差异. 实验结果显示, “U型”曲线的宽度和临界点分别与色谱动力学和热力学因素相关. 还对这两类色谱固定相的峰容量表征方法和命名提出了建议和说明. 相似文献
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Toyohide Takeuchi Budhi Oktavia Lee Wah Lim 《Analytical and bioanalytical chemistry》2009,393(4):1267-1272
Methyl-capped poly(ethylene oxide) moieties were chemically bonded to silica gel using an amine-reactive modification reagent
and evaluated as the stationary phase for ion chromatography. In this work, primary amino groups of an aminopropylsilica packing
material were reacted with methyl-PEO12-NHS ester (succinimidyl-{[N-methyl]-dodecaethyleneglycol} ester) in phosphate buffer (pH 7.0) at room temperature. The prepared poly(ethylene oxide)-bonded
stationary was evaluated for the separation of inorganic anions, and the retention behavior of inorganic anions on the prepared
stationary phase was examined. The elution order of the investigated anions was the same as that observed in common ion chromatography.
Both cations and anions of the eluent affected the retention of the analyte anions. Ion exchange was involved for the retention
of analyte anions, although the present stationary phase does not possess any discrete ion-exchange sites. The stationary
phase was applied to the separation of trace anions contained in tap water and a rock salt. 相似文献
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Numata M Kaneko T Mi Q Ye M Kawamata S Matsuo M Yarita T 《Journal of chromatography. A》2008,1210(1):68-75
In this study, a silica stationary phase modified with sulfoxide group and ammonium-salt was prepared for the separation of polychlorinated biphenyls (PCBs) from mineral oils, and its properties were investigated. Organic sulfide was attached to a diamino (primary and secondary amino) bonded silica surface by an amide bond, and the bonded sulfide groups were oxidized with periodate to afford sulfoxide groups bonded to the stationary phase. The secondary amino groups in the spacer chain were converted to ammonium-salt by the addition of hydrochloric acid. The sulfoxide group and ammonium-salt bonded stationary phase was tested for their suitability as adsorbent for SPE-type preparative short columns and for an analytical HPLC-type separation. The new stationary phase (1.2 mmol of sulfur bonded per gram) separated PCBs from mineral oils (paraffin-based transformer oils) more efficiently than previously reported stationary phases including sulfoxide group or ammonium-salt bonded ones. The quantitative chromatographic parameters for an aliphatic hydrocarbon (eicosane) and some PCB congeners also indicated strong retention of highly chlorinated biphenyls by the sulfoxide and ammonium-salt bonded silica compared with simple aminopropyl, sulfoxide group or ammonium-salt bonded ones. A cleanup procedure was established for simple determination of PCBs in mineral oil samples using sulfoxide group and ammonium-salt bonded silica packed column fractionation. The analytical method, combination of the cleanup procedure, and measurement with a GC-high resolution (magnetic sector) MS or a GC-quadrupole MS were validated using mineral oil certified reference materials. 相似文献
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Chuzo Fujimoto Akira Maekawa Yumiko Murao Kiyokatsu Jinno Tsutomu Takeichi 《Analytical sciences》2002,18(1):65-68
With the aim of preparing a stationary phase with a high shape-recognition ability for liquid chromatography, a new bonded phase was synthesized by coupling multiply dodecylamino-substituted beta-cyclodextrin (beta-CD) to 3-glycidoxypropyl-derivatized silica gel. The stationary phase prepared in this way was expected to have increased shape selectivity compared with that of conventional reversed-phase materials, due to solute interactions with the alkyl chain piles built up on the beta-CDs bonded to silica. The separation characteristics of the bonded phase were investigated using polycyclic aromatic hydrocarbons (PAHs) with different molecular shapes and compared with those of monomeric ODS and native beta-cyclodextrin-bonded phases. The newly developed stationary phase was found to be highly selective for PAHs. 相似文献
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键合纤维素(4-甲基苯甲酸酯)手性固定相的制备及其手性拆分 总被引:3,自引:0,他引:3
采用包夹聚合法,将硅小球同硅烷化试剂反应制得乙烯基硅胶,然后将该乙烯基硅胶同经十一烯酰氯、4-甲基苯甲酰氯衍生的纤维素共聚,制备出含不同官能团的聚合物包夹硅基的键合型纤维素(4-甲基苯甲酸酯)类手性固定相。分别以正己烷异丙醇、正己烷四氢呋喃为流动相,对此键合型手性固定相的手性识别能力进行了评价。为了与同类型的涂敷型纤维素(4-甲基苯甲酸酯)手性固定相作比较,合成了涂敷型纤维素(4-甲基苯甲酸酯)手性固定相。结果表明,键合型纤维素(4-甲基苯甲酸酯)手性固定相具有一定的手性识别能力,可以拆分所研究的6种手性化合物中的4种。 相似文献
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This review surveys the recent progress in the adsorbed stationary phases for capillary electrochromatography (CEC). Adsorption-based methods for preparation of stationary phase are novel approaches in CEC, which allow rapid and facile preparing stationary phases with desirable selectivity onto an open-tubular fused-silica capillary, a bare-silica or ion-exchange packed column or a monolithic silica or polymer column. A variety of adsorbing agents have been developed as adsorbed stationary phases, including ionic long-chain surfactant, protein, peptide, amino acid, charged cyclodextrin (CD), basic compound, aliphatic ionene, and ion-exchange latex particle. The adsorbed stationary phases have been applied to separation of neutral, basic and acidic organic compounds, inorganic anions and enantiomers. They have also been applied to on-line sample concentration, fast separation and study of the competitive binding of enantiomers with protein. 相似文献
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TianBaoHUANG YanShunMA YuanDeLONG YongLiangDING 《中国化学快报》2003,14(8):820-823
A novel bonded stationary phase, bidenate amide bonded stationary phase (BABSP),was prepared by reacting YWG silica gel firstly with 3-aminopropyltrimethoxysilane, then with diacid chloride. Hydrophobicity, selectivity, and especially silanophilic activity of BABSP were evaluated. The stationary phase exhibited good column efficiency and peak shape for separation of some basic solutes. 相似文献
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The solvation parameter model is used to characterize the retention properties of a 3-aminopropylsiloxane-bonded (Alltima amino), three 3-cyanopropylsiloxane-bonded (Ultrasphere CN, Ultremex-CN and Zorbax SB-CN), a spacer bonded propanediol (LiChrospher DIOL) and a multifunctional macrocyclic glycopeptide (Chirobiotic T) silica-based stationary phases with mobile phases containing 10 and 20% (v/v) methanol-water. The low retention on the polar chemically bonded stationary phases compared with alkylsiloxane-bonded silica stationary phases arises from the higher cohesion of the polar chemically bonded phases and an unfavorable phase ratio. The solvated polar chemically bonded stationary phases are considerably more hydrogen-bond acidic and dipolar/polarizable than solvated alkylsiloxane-bonded silica stationary phases. Selectivity differences are not as great among the polar chemically bonded stationary phases as they are between the polar chemically bonded phases and alkylsiloxane-bonded silica stationary phases. 相似文献
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Ionic liquids (ILs) immobilized on silica as novel high performance liquid chromatography (HPLC) stationary phases have attracted considerable attention. However, it has not been applied to protein separation. In this paper, N-methylimidazolium IL-modified silica-based stationary phase (SilprMim) was prepared and investigated as a novel multi-interaction stationary phase charged positively for protein separation. The results indicate that all of the basic proteins tested cannot be absorbed on this novel stationary phase, whereas all of the acidic proteins tested can be retained, and the baseline separation of eight kinds of acidic protein standards can be achieved when performed in reversed phase/ ion-exchange chromatography (RPLC/IEC) mode. Compared with commonly used commercial octadecylated silica (ODS) column, the novel stationary phase can show selectivity and good resolution to acidic proteins, which has a promising application in the separation and analyses of acidic proteins from the complex samples in proteomics. In addition, the chromatographic behavior of proteins, the effect of the ligand structure and the retention mechanism on this stationary phase were also investigated. 相似文献
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A variety of stationary phases were bonded to silaceous supports through an intermediate silane coupling agent, γ-glycidoxypropylsilane. Consequently, a series of ion-exchange, hydrophobic, and hydrophilic supports were prepared that are stable, withstand high mobile phase velocities, and have sufficient porosity to allow the partitioning of biopolymers. Liquid-liquid partition chromatography of proteins was achieved with phosphate buffers while the separation of a series of aromatic compounds was accomplished on the same columns with hexane.
Anion- and cation-exchange supports were found to have hemoglobin ion- exchange capacities similar to classical cross-linked dextrans while allowing separation speeds 10–20 times those of carbohydrate gel supports. 相似文献
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The chromatographic behaviour of a commercially available ion-exchange stationary phase (the Dionex IonPac CS12A column) is described for a wide range of transition and heavy metal ions with nitric acid eluents containing chloride and nitrate potassium salts. The separation selectivity was found to arise from simultaneous ion-exchange interactions and chelation with the attached carboxylic and phosphonic acid groups. These interactions were investigated by altering the ionic strength and pH of the eluent and also the column temperature. Strong affinity of the stationary phase towards heavy metal ions, in particular bismuth and the uranyl ion was observed at low pH under chelating ion-exchange conditions, with high efficiency separations of other ions including cadmium and lead being possible with short analysis times (approximately 5-15 min). Examples are given of separations obtained using 4-(2-pyridylazo)resorcinol or Arsenazo III as the post-column chromogenic reagents, demonstrating the potential versatility and utility of this stationary phase for heavy metal ion analysis. 相似文献