MSPD Extraction of Sulphonamides from Meat followed by LC Tandem MS Determination

In this paper an extraction procedure is presented for sulphonamides (SAs) in animal tissues (raw meat and meat-based baby food), based on matrix solid phase dispersion (MSPD). Determination was carried out using liquid chromatography coupled with tandem mass spectrometry. In complex matrices extraction is usually the critical step of analysis: several procedures were employed for the recovery of SAs in animal tissues, but often they entail time and solvent-consuming processes. MSPD has demonstrated itself to be a very efficient extraction technique for matrices such as animal tissues, with simplified and shortened procedures, and very low solvent consumption. This procedure allowed to obtain quantitative recovery, high selectivity improved by a cool eluant, and easy application. Good recoveries are accompanied by low RSDs and not a significative matrix effect. The MSPD procedure, coupled with LC-MS-MS analysis, provides very interesting LODs, due also to the concentration step. Validation is carried out following the EU guidelines for confirmation methods (EU Dec. 657/02).     

Tandem MS Analysis of Selenamide-Derivatized Peptide Ions

Our previous study showed that selenamide reagents such as ebselen and N-(phenylseleno)phthalimide (NPSP) can be used for selective and rapid derivatization of protein/peptide thiols in high conversion yield. This paper reports the systematic investigation of MS/MS dissociation behaviors of selenamide-derivatized peptide ions upon collision induced dissociation (CID) and electron transfer dissociation (ETD). In the positive ion mode, derivatized peptide ions exhibit tag-dependent CID dissociation pathways. For instance, ebselen-derivatized peptide ions preferentially undergo Se–S bond cleavage upon CID to produce a characteristic fragment ion, the protonated ebselen (m/z 276), which allows selective identification of thiol peptides from protein digest as well as selective detection of thiol proteins from protein mixture using precursor ion scan (PIS). In contrast, NPSP-derivatized peptide ions retain their phenylselenenyl tags during CID, which is useful in sequencing peptides and locating cysteine residues. In the negative ion CID mode, both types of tags are preferentially lost via the Se–S cleavage, analogous to the S–S bond cleavage during CID of disulfide-containing peptide anions. In consideration of the convenience in preparing selenamide-derivatized peptides and the similarity of Se–S of the tag to the S–S bond, we also examined ETD of the derivatized peptide ions to probe the mechanism for electron-based ion dissociation. Interestingly, facile cleavage of Se–S bond occurs to the peptide ions carrying either protons or alkali metal ions, while backbone cleavage to form c/z ions is severely inhibited. These results are in agreement with the Utah-Washington mechanism proposed for depicting electron-based ion dissociation processes.     

Solid state 13C NMR can distinguish between bronsted and Lewis acid adducts of amines

The solid ¹³C NMR spectra of solid complexes of phenethylamine with SnCl_4, Me₃SnCl, AlCl_3, and HCl are presented and discussed as a means of distinguishing between Bronsted and Lewis acid adducts of alkyl amines. Solution ¹³C NMR results are discussed in terms of species present in complex equilibria and their detection using the two NMR methods.     

串级质谱应用于蛋白质科学研究的实例介绍

串级质谱（MS/MS）分析是近年来迅猛发展的仪器分析技术,对于化学相关学科与其他交叉领域都起到了极大的推动作用。在使用计算机辅助解析后,常常需要进一步进行图谱指认,而这部分内容常常被化学教学所忽略。从串级质谱的原理出发,详细分析了肽段在二级质谱中可能的断裂模式与结构特性,并进一步以模型蛋白为例详细分析了串级质谱的图谱指认和峰的归属的基本原理与实用性方法,以期对教学与科研有助。     

Improved colorimetric determination of high-molecular weight hyaluronic acid from synovial fluids

Summary A method is presented depending on the metachromic shift of a carbocyanine dye (Stains-all) in the presence of high molecular weight hyaluronic acid. Although this colorimetric reaction itself is not new, progress has been made in elaborating a technique which, in most cases, does not require any pretreatment of the original biological sample (synovial fluid) except appropriate dilution. The method is relatively free of interferences by other substances. The kinetics (time-absorption modes) at defined wavelengths have been demonstrated to be a valid tool for identifying the target macromolecule. The reliability of the method has been demonstrated with equine synovial fluids and with pure hyaluronate from rooster comb as reference substance.

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Verbesserte kolorimetrische Bestimmung von hochmolekularer Hyaluronsäure aus Synovialflüssigkeit

     

Production of hyaluronic acid by Streptococcus

The effects of the addition of lysozyme and forced aeration on the rheological properties and production of hyaluronic acid by Streptococcus zooepidemicus were investigated. Lysozyme was added to the culture broth in two pulses during the exponential and stationary phases of a fermentation carried out in a rotary shaker (150 rpm), using 200 mL Erlenmeyer flasks. The effect of aeration was evaluated by feeding air into a 2.5 L fermentor at a 2 vvm rate. The effects were analyzed in terms of concentration, viscosity, viscoelasticity, and molecular weight of the hyaluronic acid produced.     

Crosslinking of hyaluronic acid with glutaraldehyde

Hyaluronic acid (HA) was chemically crosslinked with glutaraldehyde (GA) to produce water-insoluble films having low water contents when brought into contact with water. The crosslinking reaction was performed using uncrosslinked HA films in acetone–water mixtures. This method could produce water-insoluble HA films with water contents as low as 60 wt % when subjected to swelling with phosphate-buffered saline of pH 7.4 at 37°C. This 60 wt % water content was lower than any values for HA ever reported. There was an optimal HCl concentration around 0.01N for the HA crosslinking with GA in acetone—water mixtures. To get information on the crosslinking mechanism, alginic acid, which possesses hydroxyl and carboxyl groups in one molecule, similar to HA, and poly(vinyl alcohol) (PVA) and amylopectin, which possess only hydroxyl groups, were subjected to crosslinking with GA. PVA and amylopectin were also found to become water-insoluble after reaction with GA. On the basis of the infrared spectra of these crosslinked films, it was concluded that intermolecular formation of hemiacetal bonds with GA between the hydroxyl groups belonging to different HA molecules led to crosslinking. © 1997 John Wiley & Sons, Inc. J Polym Sci A: Polym Chem 35 : 3553–3559, 1997     

Tunable biohybrid hydrogels from coacervation of hyaluronic acid and PEO-based block copolymers

Accurately tuning the macroscopic properties of biopolymer-based hydrogels remains challenging due to the ill-defined molecular architecture of the natural building blocks. Here, we report a biohybrid coacervate hydrogel, combining the biocompatibility and biodegradability of naturally occurring hyaluronic acid (HA) with the tunability of a synthetic polyethylene oxide (PEO) -based ABA-triblock copolymer. Coacervation of the cationic ammonium or guanidinium-functionalized copolymer A-blocks with the anionic HA leads to hydrogel formation. Both mechanical properties and water content of the self-healing hydrogels can be controlled independently by altering the copolymer structure. By controlling the strength of the interaction between the polymer network and small-molecule cargo, both release rate and maximum release are controlled. Finally, we show that coacervation of HA and the triblock copolymer leads to increased biostability upon exposure to hyaluronidase. We envision that noncovalent crosslinking of HA hydrogels through coacervation is an attractive strategy for the facile synthesis of tunable hydrogels for biomedical applications.     

高效液相色谱串联质谱法对大米中26种三嗪类除草剂多残留的检测

建立了同时检测大米中26种三嗪类除草剂残留量的高效液相色谱串联质谱分析方法.样品经乙腈提取、Oasis HLB固相萃取柱净化,液质联用仪测定.26种三嗪类除草剂在10～1 000 μg/L范围内线性良好,相关系数为0.997 3～0.999 9.在10～100 μg/kg范围内,平均加标回收率为74%～100%,相对标准偏差为2.1%～8.8%.该方法可同时满足进出口大米中多种三嗪类除草剂残留的检验需要.     

Highly efficient syntheses of hyaluronic acid oligosaccharides

Highly efficient syntheses of hyaluronic acid oligosaccharides have been accomplished through the pre-activation based iterative one-pot strategy. A series of oligosaccharides ranging from di- to hexasaccharides were rapidly assembled using only near stoichiometric amounts of the building blocks without aglycon adjustment or purifications of intermediate oligosaccharides. Deprotection and oxidation protocols were developed for protective group removal and oxidation-state adjustment. The availability of such structurally well defined synthetic hyaluronic acid oligosaccharides will greatly facilitate the establishment of detailed structure-function relationships.     

Applications of Tandem Mass Spectrometry (MS/MS) in Protein Analysis for Biomedical Research

Mass Spectrometry (MS) allows the analysis of proteins and peptides through a variety of methods, such as Electrospray Ionization-Mass Spectrometry (ESI-MS) or Matrix-Assisted Laser Desorption Ionization-Mass Spectrometry (MALDI-MS). These methods allow identification of the mass of a protein or a peptide as intact molecules or the identification of a protein through peptide-mass fingerprinting generated upon enzymatic digestion. Tandem mass spectrometry (MS/MS) allows the fragmentation of proteins and peptides to determine the amino acid sequence of proteins (top-down and middle-down proteomics) and peptides (bottom-up proteomics). Furthermore, tandem mass spectrometry also allows the identification of post-translational modifications (PTMs) of proteins and peptides. Here, we discuss the application of MS/MS in biomedical research, indicating specific examples for the identification of proteins or peptides and their PTMs as relevant biomarkers for diagnostic and therapy.     

高效液相色谱串联质谱法对栀子中11种有机磷农药残留的检测

建立了高效液相色谱串联质谱法检测栀子中11种有机磷农药残留量的分析方法。样品以乙腈为提取溶剂,采用超声波辅助提取,经Carb/PSA固相萃取柱净化,液质联用仪测定。11种有机磷农药在125~2 000μg/L范围内线性良好,相关系数为0.9955~0.9998。在50、100、500μg/kg 3个添加水平的平均加标回收率为84%~107%,相对标准偏差为1.4%~10.9%。     

Two-dimensional spectroscopy can distinguish between decoherence and dephasing of zero-quantum coherences

Recent experiments on a variety of photosynthetic antenna systems have revealed that coherences among electronic states persist longer than previously anticipated. In an ensemble measurement, the observed dephasing of a coherent state can occur because of either disorder across the ensemble or decoherence from interactions with the bath. Distinguishing how much such disorder affects the experimentally observed dephasing rate is paramount for understanding the role that quantum coherence may play in energy transfer through these complexes. Here, we show that two-dimensional electronic spectra can distinguish between the limiting cases of homogeneous dephasing (decoherence) and inhomogeneous dephasing by examining how the quantum beat frequency changes within a cross peak. For the antenna complex LH2 isolated from Rhodobacter sphaeroides , we find that dephasing of the coherence between the B850 and B800 rings arises predominantly from inhomogeneity. In contrast, within the Fenna-Matthews-Olson (FMO) complex from Chlorobium tepidum , dephasing of the coherence between the first two excitons appears quite homogeneous. Thus, the observed dephasing rate sets an upper bound on decoherence for the LH2 complex while establishing both an upper and lower bound for the FMO complex.     

基于MALDI-TOF MS的串联富集策略在磷酸化蛋白组学中的应用

提出一种除盐-富集串联用于磷酸肽富集研究的思路。选用C18柱和铈(Ⅳ)修饰的壳聚糖材料进行脱盐实验,以制备的基于聚合物基体螯合Fe3+的亲和色谱材料为富集材料。将直接富集和串联策略应用到标准品和血清中,研究结果表明,该富集材料具有高选择性和高灵敏度(1.6 fmol),铈(Ⅳ)修饰的壳聚糖材料前提下的串联策略能明显降低样品的复杂性。相比直接富集方法,能够提高磷酸化肽的覆盖率。     

Photochemical degradation of hyaluronic acid by singlet oxygen

Changes of the rheological properties of hyaluronic acid (sodium-magnesium salt) solutions after exposure to UV radiation indicate a vigorous decrease in their viscosity, but its still strong shear rate dependence. Whereas the presence of the singlet oxygen sensitizer (anthracene-1-sulphonic acid) brings about a loss of shear dependence; the studied solutions show newtonian behavior.     

Electrical conductivity measurements of hyaluronic acid and collagen

Electrical properties of hyaluronic acid (HYA) and collagen in the temperature range from –90°C (resp. –50°C for collagen) to 40°C are described. The value and type of electrical conductivity, temperature stability, and processes arising in the investigated samples during continual stress heating are determined by the measurements of temperature dependence of the electrical conductivity (both d.c. and a.c.).     

Degradation of hyaluronic acid at the metal surface

 When the viscosity of hyaluronic acid (HA) solution was measured with a rotational visco-meter, depolymerization occurred only by leaving the sample in the cup of the viscometer, while the solution in a glass test tube at the same temperature, 37 °C, was practically stable during the entire period of investigation. As the parts where the sample solution had contact with the viscometer are stainless steel, the participation of metal ions in the depolymerization was suspected. Therefore, experiments were carried out with stainless-steel beads in the HA solution. The results show that the molecular weight of HA decreased on exposure to the beads under aerobic conditions. The addition of 10 beads to 2 ml of a 0.1% HA solution prepared with 0.2 M NaCl reduced the molecular weight to about 75% of the initial value after 4 h. The degradation rate was proportional to the surface area of the beads. However, no dissolution of metal ion was detected, and therefore, this reaction was thought to proceed at the surface of the metal. Received: 26 September 1997 Accepted: 27 November 1997     

CoM-polynomial and topological coindices of hyaluronic acid conjugates

The molecular structure of corresponding drugs can be examined using a graph theory tool called topological index to learn about their physicochemical and biological properties. Topological index considers the pair of vertices that are connected while topological coindex takes into account the pairs of vertices that are not connected. Many of these topological indices can be readily calculated using various polynomial available in literature. We employ the concept of CoM-polynomial in this study and analyze the structure of hyaluronic acid conjugated with curcumin, paclitaxel, and methotrexate to acquire it. Many conventional topological coincides such as 1st Zagreb coindex, 2nd Zagreb coindex, 2nd modified Zagreb coindex, redefined 3rd Zagreb coindex, forgotten topological coindex, Randi? coindex etc. are also generated for these drugs.