Cross‐Reactive Sensor Arrays for the Detection of Peptides in Aqueous Solution by Fluorescence Spectroscopy

A simple but powerful method for the sensing of peptides in aqueous solution has been developed. The transition‐metal complexes [PdCl₂(en)], [{RhCl₂Cp*}₂], and [{RuCl₂(p‐cymene)}₂] were combined with six different fluorescent dyes to build a cross‐reactive sensor array. The fluorescence response of the individual sensor units was based on competitive complexation reactions between the peptide analytes and the fluorescent dyes. The collective response of the sensor array in a time‐resolved fashion was used as an input for multivariate analyses. A sensor array comprised of only six metal–dye combinations was able to differentiate ten different dipeptides in buffered aqueous solution at a concentration of 50 μM . Furthermore, the cross‐reactive sensor could be used to obtain information about the identity and the quantity of the pharmacologically interesting dipeptides carnosine and homocarnosine in a complex biological matrix, such as deproteinized human blood serum. The sensor array was also able to sense longer peptides, which was demonstrated by differentiating mixtures of the nonapeptide bradykinin and the decapeptide kallidin.     

Unique selectivity windows using selective displacers/eluents and mobile phase modifiers on hydroxyapatite

A detailed study was carried out to combine the unique selectivity of ceramic hydroxyapatite (CHA) with the separation power of selective displacement chromatography. A robotic liquid handling system was employed to carry out a parallel batch screen on a displacer library made up of analogous compounds. By incorporating positively charged, metal chelating and/or hydrogen bonding groups into the design of the displacer, specific interaction sites on CHA were targeted, thus augmenting the selectivity of the separation. The effect of different mobile phase modifiers, such as phosphate, sulfate, lactate and borate, were also investigated. Important functional group moieties and trends for the design of CHA displacers were established. Selective batch separations were achieved between multiple protein pairs which were unable to be resolved using linear gradient techniques, demonstrating the applicability of this technique to multiple protein systems. The specific interaction moieties used on the selective displacer were found to dictate which protein was selectively displaced in the separation, a degree of control not possible using a mono-interaction type resin in displacement chromatography. Mobile phase modifiers were also shown to play a crucial role, augmenting the selectivity of a displacer in a synergistic fashion. Column separations were carried out using selective displacers and mobile phase modifiers identified in the batch experiments, and baseline separation of the previously unresolved protein pairs was achieved. Further, the elution order in these systems was able to be reversed while still maintaining baseline separations. This work establishes a new class of separations which combine the selectivities of multi-modal resins, displacers/eluents, and mobile phase modifiers to create unique selectivity windows unattainable using traditional modes of operation.     

Platinum Nanoflowers on Scratched Silicon by Galvanic Displacement for an Effective SALDI Substrate

We report a new and facile method for synthesizing 3D platinum nanoflowers (Pt Nfs) on a scratched silicon substrate by electroless galvanic displacement and discuss the applications of the Pt Nfs in surface‐assisted laser desorption/ionization‐mass spectrometry (SALDI‐MS). Surface scratching of n‐type silicon is essential to induce Pt Nf growth on a silicon substrate (to obtain a Pt Nf silicon hybrid plate) by the galvanic displacement reaction. The Pt Nf silicon hybrid plate showed excellent SALDI activity in terms of the efficient generation of protonated molecular ions in the absence of a citrate buffer. We propose that the acidity of the Si? OH moieties on silicon increases because of the electron‐withdrawing nature of the Pt Nfs; hence, proton transfer from the Si? OH groups to the analyte molecules is enhanced, and finally, thermal desorption of the analyte ions from the surface occurs. Signal enhancement was observed for protonated molecular ions produced from a titania nanotube array (TNA) substrate on which Pt nanoparticles had been photochemically deposited. Moreover, surface modification of the Pt Nf silicon hybrid plate by perfluorodecyltrichlorosilane (FDTS) (to obtain an FDTS‐Pt Nf silicon hybrid plate) was found to facilitate soft SALDI of labile compounds. More interestingly, the FDTS‐Pt Nf silicon hybrid plate acts 1) as a high‐affinity substrate for phosphopeptides and 2) as a SALDI substrate. The feasibility of using the FDTS‐Pt Nf silicon hybrid plate for SALDI‐MS has been demonstrated by using a β‐casein digest and various analytes, including small molecules, peptides, phosphopeptides, phospholipids, carbohydrates, and synthetic polymers. The hybridization of Pt Nfs with a scratched silicon substrate has been found to be important for achieving excellent SALDI activity.     

Nucleophilic displacements of non-racemic α-trifluoromethyl benzylic triflates

Effective protocols for the introduction of chiral α-trifluoromethyl benzyl moieties by nucleophilic displacement of enantiomerically enriched α-trifluoromethyl benzylic triflates are presented. The effects of substrate electronics, solvent polarity, temperature, and base are studied by measuring the diastereomeric or enantiomeric excesses of the displacement products formed by coupling a variety of α-trifluoromethyl benzylic triflates with a range of nucleophiles including amines, carboxylates, thiols, and malonates. Preliminary investigations to elucidate the mechanism(s) involved in the loss of stereochemical integrity at the benzylic center in the nucleophilic displacement reactions are also reported.     

Mixed retention mechanism of proteins in weak anion-exchange chromatography

Using four commercial weak anion-exchange chromatography (WAX) columns and 11 kinds of different proteins, we experimentally examined the involvement of hydrophobic interaction chromatography (HIC) mechanism in protein retention on the WAX columns. The HIC mechanism was found to operate in all four WAX columns, and each of these columns had a better resolution in the HIC mode than in the corresponding WAX mode. Detailed analysis of the molecular interactions in a chromatographic system indicated that it is impossible to completely eliminate hydrophobic interactions from a WAX column. Based on these results, it may be possible to employ a single WAX column for protein separation by exploiting mixed modes (WAX and HIC) of retention. The stoichiometric displacement theory and two linear plots were used to show that mechanism of the mixed modes of retention in the system was a combination of two kinds of interactions, i.e., nonselective interactions in the HIC mode and selective interactions in the IEC mode. The obtained U-shaped elution curve of proteins could be distinguished into four different ranges of salt concentration, which also represent four retention regions.     

流动相组成和温度对衍生化b-环糊精键合固定相拆 分四种仲醇消旋体的影响

采用3,5-二甲苯基异氰酸酯衍生化β-环糊精键合固定相, 以正己烷-脂肪醇为流动相, 研究了脂肪醇结构及其浓度、柱温对2-苯基环己醇、2-萘基环己醇、4-三氟甲基-2-苯基环己醇以及3-(1-羟乙基)香豆素等四种仲醇手性拆分行为的影响. 当流动相中醇的摩尔浓度相同时, 异构体容量因子随脂肪醇碳链的增长而减小; 柱温升高, 异构体容量因子减小, 而分离因子增大. 用溶质的计量置换保留模型(SDM-R)和热力学理论对此现象进行了解释.     

Structure–Activity Studies on the Fluorescent Indicator in a Displacement Assay for the Screening of Small Molecules Binding to RNA

A series of xanthone and thioxanthone derivatives with aminoalkoxy substituents were synthesized as fluorescent indicators for a displacement assay in the study of small‐molecule–RNA interactions. The RNA‐binding properties of these molecules were investigated in terms of the improved binding selectivity to the loop region in the RNA secondary structure relative to 2,7‐bis(2‐aminoethoxy)xanthone (X2S) by fluorimetric titration and displacement assay. An 11‐mer double‐stranded RNA and a hairpin RNA mimicking the stem loop IIB of Rev response element (RRE) RNA of HIV‐1 mRNA were used. The X2S derivatives with longer aminoalkyl substituents showed a higher affinity to the double‐stranded RNA than the parent molecule. Introduction of a methyl group on the aminoethoxy moiety of X2S effectively modulated the selectivity to the RNA secondary structure. Methyl group substitution at the C1′ position suppressed the binding to the loop regions. Substitution with two methyl groups on the amino nitrogen atom resulted in reducing the affinity to the double‐stranded region by a factor of 40 %. The effect of methyl substitution on the amino nitrogen atom was also observed for a thioxanthone derivative. Titration experiments, however, suggested that thioxanthone derivatives showed a more prominent tendency of multiple binding to RNA than xanthone derivatives. The selectivity index calculated from the affinity to the double‐stranded and loop regions suggested that the N,N‐dimethyl derivative of X2S would be suitable for the screening of small molecules binding to RRE.     

计量置换参数Z值分量的测定(英文)

 以计量置换吸附理论(SDT A)为基础,从理论上推导出计量吸附模型中表征溶质对固定相亲合势大小的参数βa值与流动相中强置换剂浓度的对数呈线性关系。计量置换模型中的参数n和q(n和q分别代表1摩尔溶剂化溶质被吸附时,从吸附剂表面和从溶质分子表面所释放出的溶剂的物质的量)是计量置换参数Z值的分量,是两个非常有用的参数,可以从这个定量关系中直接获得。推导出的方程用苯的衍生物进行了实验验证,获得了较满意的结果。将这种方法计算得到的分量值与SDT A与计量置换保留模型(SDT R)相结合的方法得到的分量值进行了比较。     

关于混合模式色谱固定相对蛋白保留特征的研究

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.     

1921年诺贝尔化学奖获得者——索迪

在探索微观元素的过程中,同位素与放射性元素位移定律的发现无疑具有里程碑式的意义。同位素的发现修正了道尔顿原子学说,元素位移规律的发现使放射化学成为一门独立的学科,为此1921年的诺贝尔化学奖授予了同位素与元素位移定律的发现者—弗雷德里克.索迪,2011年恰逢索迪逝世55周年,特写此文来纪念他对现代放射化学与核物理学做出的贡献。