气相色谱法测定芦荟及芦荟干粉中的多糖

提取芦荟及芦荟干粉样品中的多糖，将多糖水解为单糖之后衍生化反应，用气相色谱法进行分析，实验选择了最佳分离条件，以木糖醇为内标物，用OV-225与OS-138为色谱柱固定液，使衍生物得到了很好的分离，结果表明：未经水解的芦荟中含有少量的单糖以葡萄糖的形式存在，水解后的单糖主要为甘露糖和葡萄糖，这与用乙醇沉淀多糖后测定结果一致，即芦荟及芦荟干粉中的多糖主要为甘露聚糖和甘露葡聚糖。     

Oriented molecular aggregates of porphyrin-based amphiphiles and their morphology control by a boronic acid sugar interaction

Towards the morphology control of oriented porphyrin aggregates by added saccharides, three amphiphilic porphyrins bearing boronic acid groups were synthesised. Among them, an amphiphilic tetraphenylporphyrin (4) bearing two octadecyl groups at 5,10-positions and two boronic acid groups (acting as saccharide-binding sites) at 15,20-positions has been found to act as a membrane-forming amphiphile in an aqueous system. Spectroscopic (UV–Vis and CD), light-scattering, DSC and electron micrographic studies have established that in aqueous media 4 forms stable fibrous aggregates only in the presence of saccharides, which are chirally twisted by the absolute configuration of the added saccharides. This is a novel method to control the aggregate morphology by saccharides and well imitates the morphological functions of certain cell membranes, the surfaces of which are covered by saccharides.     

硼酸及其衍生物在荧光分子开关中的应用

荧光分子开关中主体和客体之间可以通过多种方式相结合，其中带有荧光团的硼酸衍生物与糖或羟基化合物的结合是通过共价键形成的。糖化学在生命科学中起着非常重要的作用，因此对糖的识别和检测在医学、细胞生物学等领域具有至关重要的意义。硼酸衍生物可以作为荧光分子开关对糖进行有效的选择性识别，因而受到科学家们的广泛关注。本文对近年来国内外相关研究成果进行了综述。     

A new naphthimidazole derivative for saccharide labeling with enhanced sensitivity in mass spectrometry detection

A series of saccharides, including maltoheptose, blood type B antigen, pullulan and the glucan of Ganoderma lucidum, are easily converted into the naphthimidazole (NAIM) derivatives in high yields by the iodine‐promoted oxidative condensation. The NAIM‐labeled saccharides, without further purification, show enhanced signals in matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS). The combined use of NAIM derivatization and MALDI‐TOFMS analysis thus provides a rapid method for identification of saccharides even in less than 1 pmol of saccharide in the sample. Characterization of the biologically active saccharides and complex polysaccharides is also achieved through the NAIM‐derivatization method. This study can be further applied to facilitate the isolation and analysis of novel saccharides. Copyright © 2009 John Wiley & Sons, Ltd.     

A Rapid Determination of Sucrose and Fructose in Biological Samples by Video Densitometry

Abstract

Simple, rapid methods which have been developed for the separation and quantification of sucrose and fructose in biological materials are presented. The method uses thin layer chromatography. The saccharides are stained with thymol-reagent on the chromatograms. The quantitative determination of red spots of saccharides are performed with a Telechrom video densitometer. The density of the spots are directly proportional to the concentration of the saccharides in a certain range. The saccharides content of the sample may be determined with a standard curve on the same chromatograms. The reproducibility of the methods are characterized by a cofficient of variation of about 4–5%     

Boronic acid based peptidic receptors for pattern-based saccharide sensing in neutral aqueous media, an application in real-life samples

The advent of the alternative sweeteners market has signaled a demand for chemosensors which target multiple saccharides and saccharide derivatives, in aqueous media at physiological pH. This demand has largely been unmet as existing molecular receptors for saccharides have generally not shown sufficient degrees of affinity and selectivity in aqueous media. A chemosensor array for saccharides and saccharide derivatives, fully operational in aqueous media at physiological pH, has been developed and is reported herein. Boronic acid based peptidic receptors, derived from a combinatorial library, served as the cross-reactive sensor elements in this array. The binding of saccharides to these receptors was assessed colorimetrically using an indicator uptake protocol in the taste-chip platform. The differential indicator uptake rates of these receptors in the presence of saccharides were exploited in order to identify patterns within the data set using linear discriminant analysis. This chemosensor array is capable of classifying disaccharides and monosaccharides as well as discriminating compounds within each saccharide group. Disaccharides have also been distinguished from closely related reduced-calorie counterparts. This linear discriminant analysis set was then employed as a training set for identifying a specific saccharide in a real-world beverage sample. The methodology developed here augurs well for use in other real-world samples involving saccharides as well as for sensing other desired analytes.     

Colorimetric sensor array–smartphone–remote server coupling system for rapid detection of saccharides in beverages

A kind of “colorimetric sensor array–smartphone–remote server” coupling system was constructed for rapid on-site testing of saccharides. First, the binding capacity between saccharides and boric acid compounds (boric acid, phenylboronic acid and 3-nitrophenylboronic acid) was studied. The binding capacity of 3-nitrophenylboronic acid was found to be the highest, followed by phenylboronic acid and boric acid. Then a small-scale colorimetric sensor array (2 × 2) of pH indicator based on affinity interaction between 3-nitrophenylboronic acid and saccharides was developed to detect 19 kinds of saccharides. A camera phone was used to acquire the array images before and after reaction, then the self-developed color discrimination software in smartphone was applied to process pictures in order to obtain the color difference image and data of analytes. The color difference data were analyzed by several methods, including principal component analysis, hierarchical cluster analysis and linear discriminant analysis. The analysis results showed that the sensor array (2 × 2) established in this paper has great discriminative capability for 19 kinds of saccharides, and the classification accuracy is as high as 100%. Nineteen different quantitative models of saccharides that showed high accuracy and precision were established based on partial least-square method. Finally, the smartphone was connected to a remote server on which the qualitative analysis and quantitative analysis models for analytes had been established. The color difference data obtained by the smartphone were uploaded to the remote server for the qualitative analysis and quantitative analysis of saccharides. The effectiveness of the “colorimetric sensor array–smartphone–remote server” coupling system in rapid on-site detection of saccharides was further verified by the spike and recovery experiments. The qualitative analysis results showed that this coupling system could distinguish all of the analytes without a mistake, and the quantitative analysis results showed that the predicted values for saccharides were close to the real values.     

Distinction between some saccharides in scattered optical sum frequency intensity images

Using an optical sum frequency (SF) microscope with visible and infrared light pulses for excitation, we have attempted to distinguish among four saccharide species. The saccharides we studied were d-glucose, amylopectin, beta-cyclodextrin, and amylose. The wavelength of the infrared light was resonant to CH vibration. Amylose showed very weak sum frequency scattering for CH vibration among the four saccharides. As for the other three saccharides, we found a big difference in the sum frequency spectra of their CH stretching vibration near 2900 cm-1, when the incident visible and infrared light pulses were p- and s-polarized, respectively. Based on these facts, we have demonstrated a distinction between these three saccharides in the scattered SF intensity images.     

Recent progress in electrochemical oxidation of saccharides at gold and copper electrodes in alkaline solutions

This article reviews the progress made in the past 10 years, on electrochemical oxidation of saccharides in alkaline media for gold and copper electrodes. The mechanism and processes associated with the electrochemical oxidation of saccharides at native and surface coated electrodes continues to be of great interest. Despite the effort and various mechanisms proposed, still the need for an electrochemically active material that understands the complexity associated with saccharides continues to increase as their detection poses a challenge for bioanalytical chemistry and liquid chromatography.     

One saccharide sensor based on the complex of the boronic acid and the monosaccharide using electrochemical impedance spectroscopy

A novel saccharide sensor based on the covalent interaction between the boronic acid and saccharides was developed. Poly (aminophenylboronic acid) (PABA) was prepared by electropolymerizing 3-aminophenylboronic acid on gold electrode surface in acidic solution. The boronic acid group of the PABA film can form covalent-bond with different saccharides, which can change the dielectric characteristics of the PABA film, and the change of the dielectric characteristic was saccharides concentration dependent. Four kinds of saccharides could be detected by using electrochemical impedance spectroscopy. Good linear relationship and high sensitivity were obtained by this method.     

Sulfation Patterns of Saccharides and Heavy Metal Ion Binding

Sulfated saccharides are an essential part of extracellular matrices, and they are involved in a large number of interactions. Sulfated saccharide matrices in organisms accumulate heavy metal ions in addition to other essential metal ions. Accumulation of heavy metal ions alters the function of the organisms and cells, resulting in severe and irreversible damage. The effect of the sulfation pattern of saccharides on heavy metal binding preferences is enigmatic because the accessibility to structurally defined sulfated saccharides is limited and because standard analytical techniques cannot be used to quantify these interactions. We developed a new strategy that combines enzymatic and chemical synthesis with surface chemistry and label-free electrochemical sensing to study the interactions between well-defined sulfated saccharides and heavy metal ions. By using these tools we showed that the sulfation pattern of hyaluronic acid governs their heavy metal ions binding preferences.     

Array-based functional screening of heparin glycans

Array methodologies have become powerful tools for interrogation of glycan-protein interactions but have critically lacked the ability to generate cell response data. Here, we report the development of a slide-based array method exemplified by measurement of activation of fibroblast growth factor signaling by heparin saccharides. Heparan sulfate-deficient Swiss 3T3 cells were overlaid onto an aminosilane-coated slide surface onto which heparin saccharides had been spotted and immobilized. The cells were transiently stimulated with FGF2 and immunofluorescence measured to assess downstream ERK1/2 phosphorylation. Activation of this signaling pathway response was restricted to cells exposed to heparin saccharides competent to activate FGF2 signaling. Differential activation of the overlaid cells by different-sized heparin saccharides was demonstrated by quantitative measurement of fluorescence intensity. This "glycobioarray" platform has significant potential as a generic tool for functional glycomics screening.     

Relative hydrophobicity/hydrophilicity of fructose, glucose, sucrose, and trehalose as probed by 1-propanol: a differential approach in solution thermodynamics

Earlier, we developed the 1-propanol probing methodology that could separately evaluate the relative hydrophobicity and hydrophilicity scale of a given sample. We applied it here to fructose, glucose, sucrose, and trehalose and evaluated the same for these saccharides. We then construct a two-dimensional map with the hydrophobicity and the hydrophilicity axes and plot the above saccharides together with other nonelectrolytes subjected earlier to the same analysis. We point out that these saccharides together with other so-called "osmolytes" that accumulate in vivo under H2O stress occupy a small specific region near H2O in this map.     

Quantitative determination of saccharides in dietary glyconutritional products by anion-exchange liquid chromatography with integrated pulsed amperometric detection

A new technique for the assay of carbohydrates is described in which separation and quantification of neutral saccharides, aminosaccharides, glycuronic acids, and disaccharides may be accomplished in less than 50 min of total run time. This method involves optimized anion-exchange liquid chromatography coupled with integrated pulse amperometric detection. Complex carbohydrates from various sources, including dietary supplements, were hydrolyzed in a dilute solution of trifluoroacetic acid, freeze-dried, and reconstituted in water containing 2-deoxygalactose as the internal standard. The solution was filtered and separated on CarboPac PA20 column. The eluted saccharides were detected by oxidation on a gold electrode with quadruple-pulsed integrated amperometry. The calibration plots for the saccharides were linear with an average correlation coefficient of 0.999. Method precision regarding peak retention time and resolution used in the peak identifications was verified. With this method, previously difficult-to-separate saccharides, such as galactosamine, glucosamine, and N-acetylglucosamine, were successfully resolved from the neutral saccharides rhamnose, arabinose, and galactose. Mannose was also resolved from xylose, and de-acetylation of aminosaccharides prior to separation was not necessary. This technique provides an accurate and efficient means to assay carbohydrates in dietary supplements, which new federal regulations will soon mandate.     

新型含苯硼酸和萘双亲化合物的合成与囊泡荧光传感器的制备

合成了含有识别基团苯硼酸和荧光基团萘的新型对-[(5-十二烷氧基-1-氧基)萘]甲基苯硼酸{p-[(5-dodecyloxy-1-oxy) naphthalene] methyl-phenylboronic acid, DNMPBA}双亲化合物; 该化合物在THF/水选择性溶剂中自组织成囊泡, 囊泡的相变温度为56.8 ℃; 当向囊泡体系加糖时, DNMPBA囊泡中的萘生色基在345 nm的荧光峰强度急剧增强; 荧光强度随添加不同糖的变化趋势为果糖>葡萄糖>麦芽糖>乙二醇. 荧光强度增强可能归因于所形成的硼酸酯减弱了DNMPBA双亲化合物中一个氧原子孤对电子对萘生色基的猝灭作用而使荧光强度重新恢复. DNMPBA囊泡与糖的相互作用导致体系荧光强度变化, 使该体系有可能应用于检测生物物质如糖的化学传感器.     

1-(2-萘基)-3-甲基-5-吡唑啉酮衍生试剂的制备及其在高效液相色谱-质谱法测定糖类化合物中的应用

以1-(2-萘基)-3-甲基-5-吡唑啉酮(NMP)作为柱前衍生试剂,建立了简单、灵敏的糖类组分的反相高效液相色谱测定方法。NMP与糖在氨为催化剂的条件下,于70 ℃下反应可获得稳定的衍生产物。在Hypersil ODS 2反相色谱柱上,实现了8种单糖的基线分离。衍生物线性相关系数均大于0.9985,检出限为0.58～1.1 pmol。利用柱后在线串联质谱的电喷雾电离正离子模式监测,获得了各组分的质谱定性及裂解规律,特别是m/z 473的特征碎片离子可作为单糖NMP衍生物的判定依据。与1-苯基-3-甲基-5-吡唑啉酮(PMP)相比,NMP对糖的衍生化具有灵敏、简单、质谱裂解规律性强、重现性好等优点。该方法用于测定油菜花粉多糖中的单糖组成,结果令人满意。     

Determination of saccharides in sake by high-performance liquid chromatography with polarized photometric detection

A high-performance liquid chromatographic method has been developed for the determination of saccharides in sake, an alcoholic beverage brewed from rice. Saccharides in sake were separated on a normal phase (carbamoyl bonded silica) column using a linear gradient elution of water in acetonitrile. Seven saccharides, glucose, maltose, isomaltose, maltotriose, panose, isomaltotriose and ethyl alpha-D-glucoside, were determined by a polarized photometric detector. Unidentified peaks suggesting saccharides with polymerization degrees over 4 were also observed. The proposed method did not require any sample clean-up treatment. As an application, saccharide compositions in various kinds of sake were compared.     

Interactions of ether groups with saccharides. Enthalpy of dilution of aqueous systems containing S-trioxane and inositol,mannitol, or cyclohexanol

Enthalpies of dilution of aqueous systems containing trioxane, and trioxane with each of mannitol, inositol and cyclohexanol have been determined at 25°C. The data have been treated in terms of the Savage-Wood additivity principle and, in conjunction with literature data on ether compounds a revised estimate of the pairwise group-interaction enthalpy for CH₂/–O– and a first estimate for CHOH/–O– are presented. Systems of saccharides and saccharides with alcohols do not conform to the Savage-Wood principle and an explanation in terms of the specific hydration properties of saccharides is offered.     

Latex-functionalized monolithic columns for the separation of carbohydrates by micro anion-exchange chromatography

A novel stationary phase for micro ion chromatography has been prepared by coating a porous poly(butyl methacrylate-co-ethylene dimethacrylate-co-2-acrylamido-2-methyl-1-propanesulfonic acid) monolith with quaternary amine-functionalized latex particles via simple electrostatic binding. This stationary phase enabled the separation of saccharides in the mobile phase with a high-pH value consisting of aqueous ammonia solution in anion-exchange mode using evaporative light scattering for detection. Effects of both porous properties of the monolithic stationary phase and chromatographic conditions on the separation ability were studied. Under optimized conditions, an efficient separation of seven saccharides was achieved in less than 10 min. The stationary phase also enables the separation of saccharides obtained by the enzymatic hydrolysis of corn starch.     

高效液相色谱法同时测定生物质乳酸发酵液中有机酸及糖类化合物

建立了高效液相色谱(HPLC)同时测定生物质乳酸发酵液中有机酸及糖类的分析方法。使用Bio-Rad Aminex HPX-87H色谱柱,以5 mmol/L的H2SO4为流动相,在柱温55 ℃,流速0.6 mL/min条件下,采用示差折光检测器进行检测。结果表明,该方法可在17 min内实现发酵液中各种有机酸和糖类化合物等的完全分离与定量,6种有机酸和3种糖类化合物在0.15～5.19 g/L范围内的线性关系良好,回归方程的线性相关系数在0.9998以上。将该法用于米根霉发酵液的检测,两个水平的加标回收率为96.91%～103.11%,相对标准偏差(n=6)为0.81%～4.61%。该法适用于微生物发酵液中多种有机酸和糖类的快速、高效分离和定量测定。