兴奋剂检测中的化学衍生化方法

本文评述了兴奋剂检测中的各种化学衍生化方法,对蛋白同化激素、麻醉镇痛剂、β-阻断剂和利尿剂分别进行了讨论。这四类药物结构各不相同,但绝大多数属于高沸点、难挥发的极性化合物,不适于直接进行GC或GC/MS分析。通过化学衍生化.不仅可增大试样的挥发性和稳定性,减小样品的极性,使原来难以进行GC分析的试样转变成适合于色谱分析的试样,而且通过衍生化还可达到改善分离效果、提高检测灵敏度的目的。     

Two-dimensional protein database of human pancreas

We report here the two-dimensional protein database of human pancreas. The proteins were analyzed by two-dimensional electrophoresis followed by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS). Totally, 302 proteins were identified, of which about 27% were enzymes with a broad range of catalytic activities. Several of these are specifically expressed in pancreas, such as pancreatic amylase, pancreatic stone protein, pancreatitis-associated protein, pancreatic lipase, pancreatic elastase, etc. Structural and cytoskeletal proteins are also strongly represented on the gels. Thus, the pancreatic proteome reflects the organ's function. This work paves the way for further studies on pancreatic protein expression in health and disease, such as diabetes and pancreatic cancer.     

Mass transport in Triton X series nonionic surfactant solutions: a new approach to solute-solvent interactions

Densities, viscosities and tracer diffusion coefficients for solutions of the nonionic surfactants Triton X-45, X-114, X-100 and X-102 in water (except for Triton X-45) and methanol, and for Triton X-100 in three water-methanol mixtures have been measured at 298.15 K and 308.15 K. The activation energy for viscous flow and the contributions to it from solute and solvent have been calculated. Comparison of the Gibbs energies of these systems shows the roles of polyoxyethylene chains of Triton X molecules in the interactions between solute and solvent, and also the effects of solvents on the inter-actions for different solutes. The viscosityB coefficients are positive for all of the surfactant solutions, and the temperature coefficients ofB are negative. In the non-micellar solutions in methanol, values ofB are small and the temperature coefficient ofB is nearly zero. In water-methanol mixtures the critical micelle concentration becomes larger as the methanol content increases up to 40% w/w; micelles are not formed at higher methanol concentrations. TheB coefficient decreases with increasing proportion of methanol in the solvent, and the temperature coefficient ofB changes from a fairly large positive value at low methanol contents to a small negative value at 80% w/w methanol.     

一种利用质谱信息进行结构特征鉴定的模式识别方法

本文提出了分段式离子系列谱编码方法,实验证明具有很好的结构表征能力。本程序主要是以模式识别技术为基础,实现了对未知物谱图的KNN分类,进而得到未知物的主要结构特征。这个程序应用于有机化合物的分类,取得了满意的结果。     

Capillary electrophoretic determination of lignin degradation products obtained by permanganate oxidation

A new capillary electrophoretic (CE) technique was developed for the separation of lignin degradation products after permanganate oxidation, yielding information about quality and quantity of various linkages in the lignin molecule. This CE method is a promising alternative to existing gas chromatographic (GC) methods. An advantage in comparison with GC is the short separation time and the fact that the oxidation products (aromatic acids) can be analyzed without derivatization. The selectivity and sensitivity of CE combined with UV detection is adequate and makes it suited for fast routine characterization of lignins. If necessary, the CE method can be coupled with electrospray ionization mass spectrometry in order to make a clear assignment of the peaks.     

Analysis of carboxylic acids by gas chromatography of acetonyl esters. Application to aromatic,dicarboxylic and higher fatty acids

Summary The gas chromatographic analysis of the acetonyl esters of aromatic, dicarboxylic and higher fatty acids is reported and discussed together with their electron impact and chemical ionisation mass spectra. An improved method for the preparation of acetonyl esters is discussed.     

Drug metabolite cluster centers-based strategy for comprehensive profiling of Neomangiferin metabolites in vivo and in vitro and network pharmacology study on anti-inflammatory mechanism

Neomangiferin (NMF) is an extremely special xanthone that could be simultaneously attributed to C-glycoside and O-glycoside with a variety of biological activities, such as anti-inflammatory, antitumor, antipyretic, and so on. So far as we know, the metabolism profiling has been insufficient until now. Herein, Drug Metabolite Cluster Centers (DMCCs)-based Strategy has been developed to profile the NMF metabolites in vivo and in vitro. Firstly, the DMCCs was proposed depending on literature-related and preliminary analysis results. Secondly, the specific metabolic rule was implemented to screen the metabolites of candidate DMCCs from the acquired Ultra High Performance Liquid Chromatography Quadrupole Exactive Orbitrap Mass Spectrometry (UHPLC-Q-Exactive Orbitrap MS) data by extracted ion chromatography (EIC) method. Thirdly, candidate metabolites were accurately and tentatively identified according to the pyrolysis law of mass spectrometry, literature reports, comparison of reference substances, and especially the diagnostic product ions (DPIs) deduced preliminarily. Finally, network pharmacology was adopted to elucidate the anti-inflammatory action mechanism of NMF on the basis of DMCCs. As a result, 3 critical metabolites including NMF, Mangiferin (MF) and Norathyriol (NA) were proposed as DMCCs, and a total of 61 NMF metabolites (NMF included) were finally screened and characterized coupled with 3 different biological sample preparation methods including solid phase extraction (SPE), acetonitrile precipitation and methanol precipitation. Among them, 32 metabolites were discovered in rat urine, 30 in rat plasma, 12 in rat liver, 9 metabolites in liver microsomes and 8 in rat faeces, respectively. Our results also illustrated that NMF primarily underwent deglucosylation, glucuronidation, methylation, sulfation, dihydroxylation and their composite reactions in vivo and in vitro. Additionally, network pharmacology analysis based on DMCCs revealed 85 common targets of disease-metabolites, and the key targets were TNF, EGFR, ESR1, PTGS2, HIF1A, IL-2, PRKCA and PRKCB. They exerted anti-inflammatory effects mainly through the pathways of inflammatory response, calcium-dependent protein kinase C activity, nitrogen metabolism, pathways in cancer and so on. In general, our study constructed a novel strategy to comprehensive elucidate the biotransformation pathways of NMF in vivo and in vitro, and provided vital reference for further understanding its anti-inflammatory action mechanism. Moreover, the established strategy could be generalized to the metabolism and action mechanism study of other natural products.     

Influence of particle diameter distribution on protein recovery in the expanded bed adsorption process

The General Rate model has been developed and solved to describe protein adsorption in an expanded bed. The model takes into account axial and local variation of particle size distribution (PSD), external and intra-particle mass transfer resistances, and dispersion in liquid phase. The influence of PSD on breakthrough profiles has been analysed. The simulation results show that for a significantly high expanded bed the lower part of the breakthrough curve profiles, calculated for local particle size distribution (LPSD) and for axial average particle size distribution (APSD) are very similar. However, the upper part of breakthrough profiles calculated for LPSD approaches inlet concentration much more slowly than those calculated for APSD. The retention times of the lower part of uptake curves calculated with average particle diameter are constantly shorter than those obtained from LPSD. For the calculation of the dynamic capacity (DC), the LPSD can be replaced by APSD for large expanded bed heights. Using breakthrough profiles calculated for average particle size, DC values are constantly underestimated.     

Elucidating Factors Manipulated the Formation of Multiply Charged Protein Homomultimeric Complexes by Electrospray Ionization

Native non‐covalently bonded protein‐protein and protein‐substrate complexes are of great interest and have been extensively studied by electrospray ionization mass spectrometry (ESI‐MS). Multiply charged protein homomultimeric complexes are shown to form by ESI‐MS. This study addresses factors that can artificially induce the formation of multiply charged protein homomultimeric complexes. Cytochrome c (Cyt c) and ubiquitin, which are monomers in solution, were found to generate (Cyt c)_mⁿ⁺ by electrospray ionization (ESI). The homomultimeric complexes were not limited to dimeric complexes but include also multiply charged trimers, tetramers, and pentamers. The observation of these homomultimeric complexes has never been revealed from a Cyt c solution at the concentration as low as 10 μM. Increasing the concentration of Cyt c enhanced the formation of (Cyt c)_mⁿ⁺ as expected; however, the protein concentration does not affect the relative intensities of monomeric and dimeric complexes. Additionally the enrichment of NH₄OH also promotes the formation of (Cyt c)_mⁿ⁺. Notably, source collision‐induced dissociations (source‐CID) of (Cyt c)_mⁿ⁺ alter the charge state distribution (CSD) and may lead to an incorrect interpretation of Cyt c conformations. Hence, extra care should be taken when using CSD to interpret the conformation of a protein derived from ESI‐MS.     

Study of the development of thermoresistance in human pancreatic carcinoma cell lines using proteome analysis

In order to find candidate proteins that are potentially associated with the thermoresistant phenotype in combination with drug resistance, we analyzed the differential protein expression in vitro in the human pancreatic cancer cell line EPP85-181-P and classical and atypical multidrug-resistant variants and their thermoresistant counterparts using proteomics. This study identifies sets of proteins that may lead to the development of thermoresistance. These results provide a fundamental basis to elucidate the molecular mechanism of thermoresistance and chemoresistance phenomena that may assist the therapy of inoperable cancers.