Gangliosides as Biomarkers of Human Brain Diseases: Trends in Discovery and Characterization by High-Performance Mass Spectrometry

Gangliosides are effective biochemical markers of brain pathologies, being also in the focus of research as potential therapeutic targets. Accurate brain ganglioside mapping is an essential requirement for correlating the specificity of their composition with a certain pathological state and establishing a well-defined set of biomarkers. Among all bioanalytical methods conceived for this purpose, mass spectrometry (MS) has developed into one of the most valuable, due to the wealth and consistency of structural information provided. In this context, the present article reviews the achievements of MS in discovery and structural analysis of gangliosides associated with severe brain pathologies. The first part is dedicated to the contributions of MS in the assessment of ganglioside composition and role in the specific neurodegenerative disorders: Alzheimer’s and Parkinson’s diseases. A large subsequent section is devoted to cephalic disorders (CD), with an emphasis on the MS of gangliosides in anencephaly, the most common and severe disease in the CD spectrum. The last part is focused on the major accomplishments of MS-based methods in the discovery of ganglioside species, which are associated with primary and secondary brain tumors and may either facilitate an early diagnosis or represent target molecules for immunotherapy oriented against brain cancers.     

气相色谱-质谱法分析歧化松香中性部分的化学组成

用皂化法将歧化松香中的酸性和中性部分分离,并由GC/MS法对中性部分的化学组成进行了分析,为进一步拓宽松脂可再生资源的用途提供了科学依据。实验考察了中性部分化学组分分析的GC和MS条件,检索对照了NIST数据库的标准图谱,结果表明,歧化松香中性部分共分离出55个组分,其中高沸点中性部分共分离出32个组分,鉴定了其中的30个组分(占歧化松香中性物质质量的80.3%,下同),主要化学成分为:脱氢枞酸甲酯、7-二氢海松酸甲酯、8(14)-二氢异海松酸甲酯、8α-四氢异海松酸甲酯、7-二氢枞酸甲酯、8-二氢海松酸甲酯、8α-四氢海松酸甲酯以及4个高沸点化合物;低沸点中性部分分离出23个组分,鉴定了其中的22个组分(占歧化松香中性物质质量的19.7%),主要是倍半萜和少量单萜化合物,系残留的歧化松节油。     

液相色谱串联质谱法分析油菜和土壤中烯草酮的残留

采用液相色谱串联三重四极杆质谱法,流动相为乙腈-水-甲酸(体积比90:10:0.1),采用多反应监测模式(MRM),测定油菜和土壤中的烯草酮残留.结果表明:添加水平分别为0.01、0.05、0.1 mg/kg时,测定回收率在90%～100%之间,相对标准偏差小于10%.     

Contribution of Capillary Zone Electrophoresis Hyphenated with Drift Tube Ion Mobility Mass Spectrometry as a Complementary Tool to Microfluidic Reversed Phase Liquid Chromatography for Antigen Discovery

The discovery of new antigens specific to multiple myeloma that could be targeted by novel immunotherapeutic approaches is currently of great interest. To this end, it is important to increase the number of proteins identified in the sample by combining different separation strategies. A capillary zone electrophoresis (CZE) method, coupled with drift tube ion mobility (DTIMS) and quadrupole time-of-flight mass spectrometry (QTOF), was developed for antigen discovery using the human myeloma cell line LP-1. This method was first optimized to obtain a maximum number of identifications. Then, its performance in terms of uniqueness of identifications was compared to data acquired by a microfluidic reverse phase liquid chromatography (RPLC) method. The orthogonality of these two approaches and the physicochemical properties of the entities identified by CZE and RPLC were evaluated. In addition, the contribution of DTIMS to CZE was investigated in terms of orthogonality as well as the ability to provide unique information. In conclusion, we believe that the combination of CZE-DTIMS-QTOF and microfluidic RPLC provides unique information in the context of antigen discovery.     

液相色谱及液质联用技术在环境分析中的应用

高效液相色谱（HPLC）是在经典液相色谱法和气相色谱法的基础上发展起来的新型分离分析技术。20世纪80、90年代后,HPLC技术发展迅速,特别是21世纪初,液相色谱与现代质谱的联机技术有了很大的发展,使HPLC的应用前景更为广阔。在全部已知的有机化合物中近80％的有机化合物属于挥发性低,易受热分解或者大分子化合物,适合于高效液相色谱进行分析。本文主要从HPLC分析方法的特点为切入点,介绍液相色谱及液质联用技术在环境分析中的应用。     

全二维气相色谱-飞行时间质谱在香精分析中的应用进展

全二维气相色谱是一种新型的功能强大的分离分析技术.介绍了全二维气相色谱-飞行时间质谱在香精香料研究领域的研究进展,该技术在香精香料的成分分析方面具有广阔的应用前景.     

气相色谱/质谱法在环境水样分析中的应用

使用树脂吸附、溶剂洗脱、GC/MS联用技术对某地一化工厂向河流中直接排放的废水、河流的上河水和下河水及地下水进行了分析和检验,检测出样品中所含的数十种有机化合物。该方法快速、灵敏,且定性定量准确,为行政管理部门实施监督和管理,工厂对排放水进行控制和处理及工厂周围居民对所饮用水的水质了解,提供了有效的途径和依据。     

Zinc Deficiency Leads to Lipid Changes in Drosophila Brain Similar to Cognitive-Impairing Drugs: An Imaging Mass Spectrometry Study

Several diseases and disorders have been suggested to be associated with zinc deficiency, especially learning and memory impairment. To have better understanding about the connection between lipid changes and cognitive impairments, we investigated the effects of a zinc-chelated diet on certain brain lipids of Drosophila melanogaster by using time-of-flight secondary ion mass spectrometry (ToF-SIMS). The data revealed that there are increases in the levels of phosphatidylcholine and phosphatidylinositol in the central brains of the zinc-deficient flies compared to the control flies. In contrast, the abundance of phosphatidylethanolamine in the brains of the zinc-deficient flies is lower. These data are consistent with that of cognitive-diminishing drugs, thus providing insight into the biological and molecular effects of zinc deficiency on the major brain lipids and opening a new treatment target for cognitive deficit in zinc deficiency.     

Native Liquid Chromatography and Mass Spectrometry to Structurally and Functionally Characterize Endo-Xylanase Proteoforms

Xylanases are of great value in various industries, including paper, food, and biorefinery. Due to their biotechnological production, these enzymes can contain a variety of post-translational modifications, which may have a profound effect on protein function. Understanding the structure–function relationship can guide the development of products with optimal performance. We have developed a workflow for the structural and functional characterization of an endo-1,4-β-xylanase (ENDO-I) produced by Aspergillus niger with and without applying thermal stress. This workflow relies on orthogonal native separation techniques to resolve proteoforms. Mass spectrometry and activity assays of separated proteoforms permitted the establishment of structure–function relationships. The separation conditions were focus on balancing efficient separation and protein functionality. We employed size exclusion chromatography (SEC) to separate ENDO-I from other co-expressed proteins. Charge variants were investigated with ion exchange chromatography (IEX) and revealed the presence of low abundant glycated variants in the temperature-stressed material. To obtain better insights into the effect on glycation on function, we enriched for these species using boronate affinity chromatography (BAC). The activity measurements showed lower activity of glycated species compared to the non-modified enzyme. Altogether, this workflow allowed in-depth structural and functional characterization of ENDO-I proteoforms.     

Comparison of Liquid Chromatography with Fluorescence Detection and Gas Chromatography/Mass Spectrometry for the Determination of Polycyclic Aromatic Hydrocarbons in Environmental Samples

Liquid chromatography (LC) with fluorescence detection and gas chromatography/mass spectrometry (GC/MS) have been compared for the determination of polycyclic aromatic hydrocarbons (PAHs) in a variety of environmental samples. Three sets of data are presented in this paper in which LC/fluorescence and GC/MS were used for the analysis of the same samples. These three data sets include the comparison of results from: (1) certification measurements for three natural matrix Standard Reference Materials (SRM's), (2) an international round robin for the determination of PAHs in air and diesel particulate samples, and (3) the analysis of four marine sediment reference materials. The results from these studies indicate that the two techniques generally provide comparable results for the measurement of PAHs in environmental samples (in the range of 0.1 to 300 ng/g), with differences in the two techniques between 5–20%. However, at low levels, anthracene and perylene are best measured using LC/fluorescence because of their selective and sensitive fluorescence detection characteristics. In contrast, GC/MS provides more accurate results for the determination of benzo[ghi]perylene because of its low fluorescence sensitivity.     

卷烟主流烟气中吡啶、喹啉和苯乙烯的GC/MS测定

采用剑桥滤片捕集卷烟主流烟气粒相物中的吡啶、喹啉和苯乙烯,用甲醇溶液萃取,萃取液经0.45μm滤膜后,气相色谱/质谱联用(GC/MS)测定分析卷烟主流烟气粒相物中的吡啶、喹啉和苯乙烯3种半挥发性物质含量,3种物质的线性范围分别为0.6~12.0、0.2~6.0、0.1~0.8μg/mL,检出限分别为0.54、0.03、0.03 ng/cig,相关系数(r2)达到0.995以上。     

Characterization and Quantification of Selenoprotein P: Challenges to Mass Spectrometry

Selenoprotein P (SELENOP) is an emerging marker of the nutritional status of selenium and of various diseases, however, its chemical characteristics still need to be investigated and methods for its accurate quantitation improved. SELENOP is unique among selenoproteins, as it contains multiple genetically encoded SeCys residues, whereas all the other characterized selenoproteins contain just one. SELENOP occurs in the form of multiple isoforms, truncated species and post-translationally modified variants which are relatively poorly characterized. The accurate quantification of SELENOP is contingent on the availability of specific primary standards and reference methods. Before recombinant SELENOP becomes available to be used as a primary standard, careful investigation of the characteristics of the SELENOP measured by electrospray MS and strict control of the recoveries at the various steps of the analytical procedures are strongly recommended. This review critically discusses the state-of-the-art of analytical approaches to the characterization and quantification of SELENOP. While immunoassays remain the standard for the determination of human and animal health status, because of their speed and simplicity, mass spectrometry techniques offer many attractive and complementary features that are highlighted and critically evaluated.     

Effects of Storage Temperature on Indica-Japonica Hybrid Rice Metabolites,Analyzed Using Liquid Chromatography and Mass Spectrometry

The Yongyou series of indica-japonica hybrid rice has excellent production potential and storage performance. However, little is known about the underlying mechanism of its storage resistance. In this study, Yongyou 1540 rice (Oryza sativa cv. yongyou 1540) was stored at different temperatures, and the storability was validated though measuring nutritional components and apparent change. In addition, a broad-targeted metabolomic approach coupled with liquid chromatography-mass spectrometry was applied to analyze the metabolite changes. The study found that under high temperature storage conditions (35 °C), Yongyou 1540 was not significantly worse in terms of fatty acid value, whiteness value, and changes in electron microscope profile. A total of 19 key differential metabolites were screened, and lipid metabolites related to palmitoleic acid were found to affect the aging of rice. At the same time, two substances, guanosine 3′,5′-cyclophosphate and pipecolic acid, were beneficial to enhance the resistance of rice under harsh storage conditions, thereby delaying the deterioration of its quality and maintaining its quality. Significant regulation of galactose metabolism, alanine, aspartate and glutamate metabolism, butyrate metabolism, and arginine and proline metabolism pathways were probably responsible for the good storage capacity of Yongyou 1540.     

Parallel Reaction Monitoring: A Targeted Experiment Performed Using High Resolution and High Mass Accuracy Mass Spectrometry

The parallel reaction monitoring (PRM) assay has emerged as an alternative method of targeted quantification. The PRM assay is performed in a high resolution and high mass accuracy mode on a mass spectrometer. This review presents the features that make PRM a highly specific and selective method for targeted quantification using quadrupole-Orbitrap hybrid instruments. In addition, this review discusses the label-based and label-free methods of quantification that can be performed with the targeted approach.     

液相色谱串联质谱与通过式固相萃取结合测定人体血液和血清中的伏立康唑和卡马西平

采用通过式固相萃取建立高效液相色谱-串联质谱(HPLC-MS/MS)快速测定人体血液和血清中伏立康唑和卡马西平的方法.样品经蛋白沉淀后过PRiME HLB通过式固相萃取柱净化,以Poroshell 120 EC-C18(4.6 mm×50 mm,2.7μm)为色谱柱,0.01 mol/L醋酸铵水溶液-乙腈(含0.1％甲...     

Study of the Fate of Lipophilic Wood Extractives During Acid Sulphite Pulping Process by Ultrasonic Solid-Liquid Extraction and Gas Chromatography Mass Spectrometry

Abstract

Lipophilic extractives in dissolving pulp are still a big concern for pulp industries due to their detrimental effect on the quality of pulp. Since lipophilic extractives remaining in pulp are trapped in low-accessible fiber structures, effective extraction methods need to be established for studying their fate in the process. A simple, ultrasonic, solid liquid extraction established in this study was found to be more effective in capturing extractive residues in all “elemental chlorine-free” bleaching stages. Its efficiency was dependent on the extraction temperature, time, and solvent composition. A linearly correlated removal pattern of sterols and unsaturated fatty acids by the bleaching sequence was observed. It was further found that the bleaching combination did not fully remove lipophilic extractives. This study suggests an opportunity to explore ultrasonication-based extraction as an effective extraction procedure prior to gas chromatography mass spectrometry analysis for monitoring extractives in pulp.     

固相微萃取-气相色谱串联两级质谱法测定水中10种藻类代谢产物

文中建立了固相微萃取-气相色谱串联两级质谱法测定地表水中的10种藻类代谢产物的含量,研究了萃取时间、萃取温度、萃取剂量、萃取纤维等预处理参数对嗅味物质含量的影响,同时对不同规格的色谱柱进行分离效果比较。结果表明,在萃取时间为25 min、萃取温度为65℃、氯化钠含量为1.7 g的测定条件下,采用50/30μm DVB/CAR/PDMS萃取头进行预处理,HP-5MS色谱柱进行分离,质谱检测器进行测定,可以使检测结果达到最优效果。方法的质量浓度均达到5.00～100.00 ng/L,工作曲线回归方程的相关系数R²>0.995;方法检出限(2S/N)为0.14～1.00 ng/L。在纯水加标量为20.00、50.00、80.00 ng/L时,平均回收率为92.0%～109.0%,精密度RSD为0.4%～8.9%,实际水样加标的平均回收率为83.9%～101.8%。该方法检测浓度数量级在纳克级,灵敏度较固相萃取-气相色谱-质谱法提高1 000倍,更适合地表水和饮用水中低浓度藻类代谢产物的检测要求。