同步导数荧光光谱法测定食品中3种芳香族氨基酸

提出了同步导数荧光光谱法同时测定食品中色氨酸、酪氨酸和苯丙氨酸的方法。试验表明:为排除干扰,色氨酸、酪氨酸和苯丙氨酸的特征同步导数荧光波长宜选定279,224,256 nm,波长差Δλ为55,70,25 nm条件下进行同步扫描。3种芳香族氨基酸的浓度分别在一定范围内与其导数荧光强度呈线性关系,检出限（3s/k）分别为6.3×10^-9,1.6×10^-8,4.0×10^-7mol·L^-1。方法用于蜂蜜、羊奶粉和啤酒样品中3种芳香族氨基酸的测定,回收率在92.3%～103%之间。     

C18柱串联冠醚手性柱高效分离3种芳香族氨基酸对映体

将C18柱与手性冠醚柱串联,建立了一种反相高效液相色谱法用于3种芳香族氨基酸对映体同时拆分的方法.考察了反相色谱流动相的组成、pH值、柱温、流速对对映体拆分的影响.实验结果表明,当流动相为HClO4-乙睛溶液(86:14,V/V,pH 2.0)、柱温20℃、流速0.4 mL/min时,3种氨基酸对映体可获得基线分离.进一步对比了C18柱、冠醚手性柱和串联顺序不同的4种分离模式,结果表明,C18柱不能拆分氨基酸对映体,仅能分离不同种类氨基酸;冠醚手性柱可分离氨基酸映体,但不同种类氨基酸色谱峰出现重叠;串联模式能实现3种氨基酸对映体的基线分离,实现双柱优势互补,而串联顺序对分离影响不大,仅影响色谱峰的峰形.     

制备气相色谱在挥发性成分分离中的应用研究进展北大核心CSCD

分析型气相色谱仪对低沸点易挥发的有机化合物展现出优异的分离性能,通过在其色谱柱末端加装馏分收集装置,建立了现代制备气相色谱(Prep GC)技术,该技术可用于挥发性成分的快速分离富集。制备气相色谱仪是由分析型气相色谱仪改装而来,其进样系统、分离系统、检测系统、馏分收集系统也在不断地优化升级,以提高目标化合物的回收效率和纯度。Prep GC与现代波谱技术(如紫外可见吸收光谱、红外吸收光谱、拉曼光谱、质谱、X射线衍射、核磁共振波谱)耦合,可对分离富集得到的目标化合物的结构进行精准确证。近年来,与Prep GC在各种挥发性成分分离中的应用相关的报道逐渐增多,展现出良好的应用前景。然而,Prep GC在分离过程中也存在着无法制备热敏性化合物、分离成本高、易引入外源性污染等问题。该文根据近年来国内外研究学者的相关研究工作,对制备气相色谱仪的结构及其在精油单体化合物、昆虫信息素、食品和植物挥发性成分、地质生物标志物及持久性环境污染物的分离等领域的应用研究进展进行综述。最后,还对Prep GC在挥发性成分分离中的应用进行了总结与展望,旨在为拓展Prep GC应用领域提供参考。     

高效毛细管电泳法在芳香族手性氨基酸与年龄的关系研究中的应用

建立了芳香族手性氨基酸的高效毛细管电泳的分离测定方法,在最佳实验条件下,测定了平均年龄为27±2和74±4岁的两组健康者血浆中D、L-苯丙氨酸、色氨酸和酪氨酸的含量.统计分析表明,血浆中芳香族手性氨基酸的含量与人类年龄之间具有一定关系.     

高效毛细管电泳地在芳香族手性氨基酸与年龄的关系研究中的应用

建立了芳香族手性氨基酸的高效毛细管电泳的分离测定方法，在最佳实验条件下，测定了平均年为２７±２和７４±４岁的两组健康血浆中Ｄ、Ｌ－苯丙氨酸、色氨酸和酷物含量。统计分析表明，血浆中蔟时族手性氨基酸的含量和人类年龄之间具有一定关系。     

硅酸铜分离微生物发酵液中的氨基酸

采用硅酸铜对微生物发酵液中的氨基酸进行了分离研究.通过静态吸附试验考察了硅酸锌和硅酸铜对微生物发酵液中氨基酸的吸附影响.结果表明,硅酸铜比硅酸锌更适合分离发酵液中的氨基酸,并确定了以硅酸铜为填料的分离条件:上柱发酵液pH=2,流速0.2BV/h,洗脱液为1mol/L的NH4Cl.在该条件下能够一次性将3种不同的氨基酸较好的分离.     

腌制食品中N-亚硝胺类化合物分析方法的研究进展北大核心CSCD

本文简要介绍了腌制食品中N-亚硝胺类化合物的种类及特性,重点综述了气相色谱法、气相色谱-质谱法、液相色谱法、液相色谱-质谱法和胶束电动毛细管色谱法等技术在腌制食品中N-亚硝胺类化合物分析中的应用进展(引用文献39篇)。     

高效液相色谱法测定地骨皮中18种氨基酸北大核心CSCD

采用高效液相色谱法测定地骨皮中18种氨基酸的含量。地骨皮样品用10-乙基吖啶酮-2-磺酰氯为柱前衍生试剂进行衍生化。以Akasil-C18色谱柱为固定相,用乙腈(5+95)溶液和乙腈(95+5)溶液以不同比例混合的溶液为流动相进行梯度洗脱,用荧光检测器测定。18种氨基酸的线性范围均为32fmol^100pmol,检出限(3S/N)在2.06~8.65fmol之间。加标回收率在96.4%~107%之间,测定值的相对标准偏差(n=6)在均小于4.0%。     

高效液相色谱法快速测定血清中的芳香族氨基酸

采用高效液相色谱-紫外检测法分离测定了血清中的芳香族氨基酸。采用的色谱柱为Waters Nova-Pak C18柱(4 μm,150 mm×3.9 mm i.d.),流动相为乙腈-水(体积比为6∶94,pH 3.4)溶液,流速为1.0 mL/min,检测波长为215 nm。血清标本经5%(体积分数)高氯酸溶液去除蛋白质后取上清液直接进样,10 min内完成测定。探讨了流动相的pH及其有机相的比例、蛋白质沉淀剂以及检测波长等因素对分离度和灵敏度的影响。考察了其他10余种氨基酸、多巴胺类等物质对目标组分检测的     

毛细管电泳-质谱技术在手性化合物分离分析中的研究进展北大核心CSCD

目前使用的绝大多数药物为手性化合物,它们具有相似的物理和化学性质,但药理活性不同,且常以外消旋混合物的形式存在,因此对手性化合物的分离在生物、环境、食品和医药等领域一直备受关注。与广泛使用的液相色谱-质谱(LC-MS)相比,毛细管电泳-质谱(CE-MS)作为一种新型分离分析技术,具有分离效率高、样品和试剂消耗量低、选择性高和分离模式多样化等诸多优势,已经发展成为手性分析领域中有广阔应用前景的分析方法之一。CE-MS结合了CE的高分离效率和低样品消耗以及MS的高灵敏度和强结构解析能力,在蛋白质组学和代谢组学等领域发挥了重要作用。CE杰出的手性拆分能力与MS优势的结合,亦使CE-MS成为实现手性化合物高效分离分析的完美组合。在过去的十几年里,基于不同CE-MS分离模式的高性能手性分析体系层出不穷,如电动色谱-质谱(EKC-MS)、胶束电动色谱-质谱(MEKC-MS)和毛细管电色谱-质谱(CEC-MS)等,并成功应用于医药、生物、食品和环境科学等领域的手性化合物分析。该文主要综述了2011~2021年,CE-MS在手性化合物分析领域的技术、手性选择剂(如改性环糊精和聚合物表面活性剂等)的使用以及在医药等领域应用方面的研究进展,并讨论了不同手性分析模式的局限性,为未来的CE-MS手性分离分析技术发展及应用提供借鉴。     

Enantiomeric separation of unusual secondary aromatic amino acids

Summary High-performance liquid chromatographic and gas chromatographic methods were developed for the separation of unusual secondary aromatic amino acids. Amino acids containing 1,2,3,4-tetrahydroisoquinoline, 1,2,3,4-tetrahydronorharmane-1-carboxylic acid and 1,2,3,4-tetrahydro-3-carboxy-2-carboline moieties were synthetized in racemic or chiral forms. The high-performance liquid chromatography was carried out either on a teicoplanin-containing chiral stationary phase or on an achiral C₁₈ column. In the latter case the diastereomers of the amino acids formed by precolumn derivatization with the chiral reagents 2,3,4,6-tetra-O-acetyl-β-D-glucopyranosyl isothiocyanate or 1-fluoro-2,4-dinitrophenyl-5-L-alanine amide were separated. The gas chromatographic analyses were based on separation on a Chirasil-L-Val column. Presented at: Balaton Symposium on High-Performance Separation Methods, Siófok, Hungary, September 3–5, 1997     

Chiral separation of halogenated amino acids by ligand-exchange capillary electrophoresis

The chiral separation of halogenated amino acids by ligand-exchange CE is described. Halogenated amino acids attracted increasing interest in recent years because of their physiological activities. Different chiral selectors, as there are L-4-hydroxyproline, L-histidine, and N-alkyl derivatives of L-4-hydroxyproline in form of their copper(II) complexes, are compared for their chiral recognition ability for halogenated amino acids. The influence of various parameters, such as selector concentration, pH, organic modifier, and field strength, on the resolution was investigated. All halogenated amino acids investigated were baseline-separated under optimized conditions.     

土壤中氨基酸分析方法的研究进展

本文对土壤中氨基酸的分析方法做了较详细的评述,并针对土壤氨基酸的特点,就土壤样品的前处理(主要包括水解和纯化)及目前常用的色谱分离分析方法进行了总结(主要包括柱后衍生阳离子交换色谱、气相色谱、柱前衍生反相高效液相色谱、阴离子交换色谱-积分脉冲安培检测等方法)。     

High-performance liquid chromatographic separation of stereoisomers of N-phthaloyl-protected amino acids and dipeptidomimetics

The stereoisomers of N-phthaloyl-protected amino acids and dipeptidomimetics were separated on macrocyclic glycopeptide and cellulose-based chiral stationary phases (CSPs) in the RP and polar-ionic modes. The effects of the organic modifier, the mobile phase composition, and the pH on the separations were investigated. Optimization of these separations was achieved through variation of the mobile-phase additive combinations. The elution sequence was determined for some of the samples. A practical application for the monitoring of the reaction conditions for N-phthaloylation of (S)-Phe was demonstrated.     

Synthesis of phosphinic and phosphonic analogs of aromatic amino acids

The reaction of aryl and aralkyl aldoximes with hypophosphorous acid resulted in aminophosphinic acids, which were oxidized into the corresponding aminophosphonic acids. Translated fromIzvestiya Akademii Nauk. Seriya Khimicheskaya, No. 1, pp. 137–140, January, 1997.     

Reversal of the enantiomeric elution order of some aromatic amino acids using reversed-phase chromatographic supports coated with the teicoplanin chiral selector

In this paper, two chiral stationary phases were prepared by coating the surface of both C8 and C18 high-performance liquid chromatography (HPLC) supports with the teicoplanin chiral selector. The hydrophobic C11 acyl side chain, attached to the d-glucosamine group of teicoplanin, served as anchor moiety for the immobilization of the chiral selector on the apolar support material. The retention and enantioselectivity of these coated stationary phases were studied using some aromatic amino acids as probe solutes and an aqueous solution as mobile phase. It was found that the enantiomer elution order on the modified C8 and C18 stationary phases was reversed (l > d) relatively to that classically observed with a teicoplanin covalently immobilized on a silica support (d > l). Such a dynamic coating on the reversed-phase supports was found to be of interest since the apparent enantioselectivity was not significantly changed by the use during an extended period of time or following a long-term storage of the columns.     

奎宁-冠醚组合型手性固定相直接拆分氨基酸的机理

合成了一种新型奎宁-冠醚组合型手性固定相(QN-CR CSP)并用于氨基酸手性对映体的直接拆分,该固定相对12种氨基酸对映体有良好的手性拆分能力。基于氨基酸手性识别中离子交换和络合的协同作用,建立了一种新型的等温吸附模型。通过迎头特殊点洗脱法(FACP)测定色氨酸(Trp)在不同金属离子添加剂条件下的等温吸附线,验证了模型的合理性。流动相中的Li⁺、Na⁺、K⁺等金属离子与氨基酸竞争固定相中的冠醚络合位点,随着金属离子与冠醚的络合作用力和络合吸附平衡常数增大,固定相对Trp的手性拆分能力下降。该模型的建立对理解氨基酸在此类固定相中的手性保留行为以及固定相结构的进一步优化具有重要意义。     

Preparation of anhydrochymotrypsin diol silica as selective adsorbent for affinity chromatography of peptides with aromatic amino acids at C-termini

Summary Anhydrochymotrypsin (AHC), a catalytically inert derivative of chymotrypsin in which the serine-residue active site has been converted chemically to a dehydroalanine residue, was immobilized on diol silica by activation with trifluoroethanesulfonyl chloride. A AHC-diol-silica column was used for high-performance affinity chromatographic separation of peptides with aromatic amino acids at their C-termini from other peptides. Faster separations were achieved.     

Trace analysis of oxidized, nitrated, and chlorinated aromatic amino acids by capillary electrophoresis with electroosmotic flow modification allowing large-volume sample stacking

A capillary electrophoresis method has been developed for the simultaneous analysis of the oxidized, nitrated, and chlorinated aromatic amino acids, as well as their parent compounds. These modifications of the aromatic amino acids in proteins or free form are induced by the attack of reactive, mainly free radical species generated during cell stress, and these stable products may serve as biomarkers of cell damage. The analytes tyrosine, phenylalanine, dihydroxyphenylalanine, tryptophan, 3-nitrotyrosine, 3-chlorotyrosine, ortho-tyrosine, meta-tyrosine, 3-hydroxyphenylacetic acid (internal standard 1), and alpha-methyltyrosine (internal standard 2) were separated in their anionic forms in alkaline borate buffer. The polyamine spermine was used as electroosmotic flow (EOF) modifier. Adsorbing to the capillary wall, spermine can either suppress or even reverse the EOF depending on its concentration and the pH. The effects of the pH of the separation buffer, the spermine concentration, the temperature, and the applied field strength on the separation were examined. The modified aromatic amino acids are present in biological fluids in a much lower concentration than their parent compounds, thus high detection sensitivity of the analytical method is required. To achieve good detection sensitivity, field-amplified sample stacking of large injection volumes was applied. Omitting polyamine from the sample buffer allowed local reversal of the EOF, thus removal of the low conductivity sample buffer at the capillary inlet. In this way, 100% of the capillary to the detection window could be filled with the sample, and the detection limits achieved for the modified aromatic amino acids were in the range of 2.5-10 nM.     

Overview of chromatographic and electrophoretic methods for the determination of branched-chain amino acids

The significance of branched-chain amino acids in diseases was clearly shown over the years. This review aims to describe the available techniques for their analytical determination. The article provides examples of the use of various analytical methods. The methods are divided into two categories: derivatization and non-derivatization approaches. Separation is achieved through different chromatography or capillary electrophoresis techniques and can be combined with different detectors such as flame ionization, ultraviolet, fluorescence, and mass spectrometry. It compares the application of various derivatization reagents or detection as such for different detectors.