组合蒸馏技术用于气相色谱分析的样品前处理

研究了一种组合蒸馏方法的样品前处理技术,并将其研制成样品预处理装置,运用载体、携带体和组合蒸馏技术实现待测样品中微量成分的提取、分离和富集。将分子蒸馏收集端的冷端接收器设计为可更换结构,避免了样品的交叉污染问题。在乙酸乙酯中添加4种有机磷农残标样作为评价样品,所研制的装置能自动将农残组分富集到2 mL邻苯二甲酸二甲酯中。利用气相色谱分析的结果,对该样品前处理装置的分离富集参数进行了优化。实验结果表明,该方法能完全除去蛋白、脂肪、色素等对气相色谱分析有干扰的组分;能将待测微量组分的浓度富集150倍以上;回收率为90%～117%,相对标准偏差为1.3%~10%;最小检出浓度为1 ng/mL;可以处理高于100 g的样品量;可以得到几十毫克至上百毫克的目标组分;有机试剂的回收利用率大于95%;操作简单方便。用模拟样品做了实际应用实验。该技术适合沸程为120～600 ℃组分的富集。所研制的装置适用于大量样品中痕量组分的半制备级富集。     

A Rapid Sample Preparation Method of Adriamycin from Plasma for HPLC Analysis

A rapid and comprehensive sample preparation method used to extract adriamycin from plasma has been investigated. The samples were passed through an Adsorbex extraction column with an elution solvent. The eluate was directly analyzed by reversed-phase high performance liguid chromatography (HPLC) with fluorescence detector. This solid/liquid extraction method is simpler than the conventional liquid/liquid extraction method. Different elution solvents were used to extract the adriamycin from samples with and without serum. The mobile phase was found to be the optimal elution solvent: 5 mM orthophosphoric acid 62.5% in methanol; acetonitrile; isopropanol = 15:15:7.5, pH 3.2. This method has the merits of better recovery (98%), higher reproducibility, and reguires shorter time and consumes less solvent.     

基于不同裂解液的蛋白质组样品制备方法的比较研究

蛋白质组学研究所面临的首要问题就是蛋白质组的样品制备,而不同的样品制备方案针对细胞、组织等样品的裂解能力,以及对不同种类蛋白的溶解能力会有很大的差异.本研究以培养的293T细胞为对象,利用质谱分析鉴定蛋白的方法,比较了采用3种常用的蛋白质组样品制备方法(Triton X-100法、尿素法和TRIzol法)所提取蛋白种类的差异,并利用生物信息学工具对所得数据进行了分析.结果表明,Triton X-100法和尿素法所提取蛋白的鉴定数目接近,而TRIzol法所提取蛋白的鉴定数目比二者减少约8％.从所鉴定的蛋白种类来看,3种方法相差较大,仅约有32％的蛋白为3种方法所共有,生物学功能也不尽相同.本研究为评价各种蛋白质组样品制备方法提供了一种快速、有效、全面的研究方法.     

Evaluation of Sample Preparation Strategies for Human Milk and Plasma Proteomics

Sample preparation is the most critical step in proteomics as it directly affects the subset of proteins and peptides that can be reliably identified and quantified. Although a variety of efficient and reproducible sample preparation strategies have been developed, their applicability and efficacy depends much on the biological sample. Here, three approaches were evaluated for the human milk and plasma proteomes. Protein extracts were digested either in an ultrafiltration unit (filter-aided sample preparation, FASP) or in-solution (ISD). ISD samples were desalted by solid-phase extraction prior to nRPC-ESI-MS/MS. Additionally, milk and plasma samples were directly digested by FASP without prior protein precipitation. Each strategy provided inherent advantages and disadvantages for milk and plasma. FASP appeared to be the most time efficient procedure with a low miscleavage rate when used for a biological sample aliquot, but quantitation was less reproducible. A prior protein precipitation step improved the quantitation by FASP due to significantly higher peak areas for plasma and a much better reproducibility for milk. Moreover, the miscleavage rate for milk, the identification rate for plasma, and the carbamidomethylation efficiency were improved. In contrast, ISD of both milk and plasma resulted in higher miscleavage rates and is therefore less suitable for targeted proteomics.     

键合反相硅胶的简单制备方法

用柱层析分离天然有机物时，经常遇到的困难是：尽管选用了各种各样的淋洗剂体系，还是不能得到满意的分离效果。据笔者经验，这时若变换固定相（如将硅胶换成氧化铝、聚酰胺、硅藻土或采用反相键合硅胶等），可能会得到满意的分离效果。可见，丰富固定相的种类对有机化合...     

XPS分析固体粉末时的样品制备法研究

用XPS分析固体粉末样品时通常将粉末直接撒在双面胶带上进行测试.这种方法粉末容易脱落,并导致仪器污染或损坏.本文介绍一种将粉末样品铺在胶带上压成薄层进行测试的制样方法.后一种方法不需模具,简单快捷,实验表明它不仅有利于保护仪器,而且有利于提高XPS分析的灵敏度和分辨率.     

微生物代谢组学的样品前处理

微生物代谢组学是指全面分析(定性和定量)细胞生长或生产周期某一时刻细胞内和细胞周围的所有低分子量代谢物.微生物代谢组学研究需要可靠和重现地分析细胞内较宽动力学浓度范围(nmol～mmol)、化学功能各异的代谢产物,因此,需要对从生物量培养、灭活和代谢产物的提取到代谢物的定量分析这整个实验过程提出耐用、可重现和可靠的实验方案.本文介绍了灭活过程中细胞内代谢产物泄漏的处理方法,较为详细地论述了通过灭活草案捕获代谢反应活性方法和代谢物提取方法的优化,并对微生物代谢组学样品前处理的目前发展趋势提出初步见解.     

X射线荧光分析中熔融制样条件的研究

本文研究了熔融制样时熔融温度、熔融时间和脱模剂的加入量对分析结果的影响。研究结果表明,随着熔融温度的升高和熔融时间的加长,分析结果的总值将随之增大。相反,脱模剂量的增加会使分析结果降低。通过对熔融样品时产生的升华物的研究,发现在熔触过程中,四硼酸锂比样品以更大的比例逸出熔融体,从而造成了样品在分析圆片中的相对浓缩。而且在高温熔融时,钾和钠比样品中的其他元素例如硅、铝、铁、钛、钙、镁等更易于逸失。制样条件的不同引起样品和熔剂逸失的比例会有变化,它直接影响测定的结果,这证明了在X射线荧光光谱分析中保持制样条件一致的重要性。     

赖氨酸C端内切酶/胰蛋白酶顺序酶切在蛋白质组学样本制备中的评估

采用胰蛋白酶(Trypsin)单独酶切与不同酶量的赖氨酸C端内切酶(Lys-C/trypsin)顺序酶切两种方法,对293T细胞全蛋白样本进行酶解消化,系统评估Lys-C/trypsin顺序酶切与Trypsin单一酶切在蛋白质组学样本制备中的差别.实验结果表明,Lys-C/trypsin顺序酶切不仅能显著提高肽段和蛋白质的鉴定数目,同时降低遗漏K酶切位点的数目及比例,而且得到的肽段长度有利于质谱鉴定,蛋白质覆盖率明显提升.通过对酶的用量进行优化对比,最终确定了Lys-C/trypsin顺序酶切时酶的合理用量.本研究结果对提高蛋白质组学样本的制备质量以及蛋白质的序列鉴定覆盖度具有指导意义.     

GC-MS测定中的小体积样品浓缩技术

在GC-MS仪器分析中,采用窄孔毛细管分析柱(0.25 mm id/0.32.mm id),能引进样品的柱容量很小(约20～60μL气体),为了有效地完成进样过程并获得较高分辨的测定结果(离子色谱图),通常采用的技术主要有柱前"冷聚焦"、柱前分流等,以纯化和减少进样体积,确保进样过程与GC-MS系统的载气流量匹配.     

Modern Methods of Sample Preparation for GC Analysis

Today, a wide variety of techniques is available for the preparation of (semi-) solid, liquid and gaseous samples, prior to their instrumental analysis by means of capillary gas chromatography (GC) or, increasingly, comprehensive two-dimensional GC (GC × GC). In the past two decades, a large number of ‘modern’ sample-preparation techniques has been introduced, which have partly superseded their ‘classical’ counterparts. These novel techniques include off-line and on-line (sometimes semi- or fully automated) procedures, and exhaustive extraction as well as equilibrium techniques. In order to improve overall performance, aspects such as essentially organic solvent-less approaches, large-volume injection and miniaturization receive increasing attention. In most recent applications, mass spectrometric or element-selective detection have been used. The present review discusses the advantages and disadvantages, and relative performance, of most of the modern sample-preparation techniques and cites a number of illustrative applications for each of them.     

A Novel Method of the Preparation ofChemically Modified Electrodes

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一个制备脱氧核糖核酸修饰电极的简便方法

金电极;聚赖氨酸;DNA;一个制备脱氧核糖核酸修饰电极的简便方法     

一种用于食品中二氧化硫快速测定的样品前处理方法

提出了一种采用半微量蒸馏-半导体制冷技术的食品中二氧化硫快速提取的新方法, 研制出了可在15 min内完成二氧化硫提取的食品检测快速蒸馏提取装置. 采用本装置,  无需冷凝水和含汞吸收剂即可实现对样品中二氧化硫的蒸馏提取. 考察了蒸馏液酸度、   蒸馏液体积、 馏分收集体积和蒸馏提取时间对二氧化硫提取效率的影响. 研究结果表明,  采用该方法在30 min内即可完成对食品中二氧化硫的快速定量测定.     

Angiopoietin-1基因修饰的骨髓间充质干细胞(MSC)的蛋白质组学方法分析

血管生成素(angiopoietin,Ang)是近来发现的促血管生成的细胞因子,其家族成员包括Angl、Ang2、Ang3、Ang4.     

Dried Blood Spot Proteomics: Surface Extraction of Endogenous Proteins Coupled with Automated Sample Preparation and Mass Spectrometry Analysis

Dried blood spots offer many advantages as a sample format including ease and safety of transport and handling. To date, the majority of mass spectrometry analyses of dried blood spots have focused on small molecules or hemoglobin. However, dried blood spots are a potentially rich source of protein biomarkers, an area that has been overlooked. To address this issue, we have applied an untargeted bottom-up proteomics approach to the analysis of dried blood spots. We present an automated and integrated method for extraction of endogenous proteins from the surface of dried blood spots and sample preparation via trypsin digestion by use of the Advion Biosciences Triversa Nanomate robotic platform. Liquid chromatography tandem mass spectrometry of the resulting digests enabled identification of 120 proteins from a single dried blood spot. The proteins identified cross a concentration range of four orders of magnitude. The method is evaluated and the results discussed in terms of the proteins identified and their potential use as biomarkers in screening programs.

戊二醛合成跟踪分析

用气相色谱法对液 液相转移反应萃取法合成戊二醛（ＧＡ）反应进行跟踪分析,采用φ３ｍｍ×４ｍ［ｍ（ＳＥ ３０）∶ｍ（硅烷化１０１白色担体）＝１５∶１００］＋φ３ｍｍ×２ｍ［ｍ（ＰＥＧ ２０Ｍ）∶ｍ（硅烷化１０１白色担体）＝１０∶１００］不锈钢柱,双柱双氢焰程序升温,升温程序为７０℃（４ｍｉｎ）１０℃／ｍｉｎ１４０℃（６ｍｉｎ）。Ｎ２４５ｍＬ／ｍｉｎ,Ｈ２６５ｍＬ／ｍｉｎ,空气３００ｍＬ／ｍｉｎ,汽化温度１８５℃,检测温度１８５℃。所建立的方法能较好地分离环戊烯、溶剂、氧化中间产物、ＧＡ、内标、氧化副产物。     

A Simple High-Throughput Field Sample Preparation Method Based on Matrix-Induced Sugaring-Out for the Simultaneous Determination of 5-Hydroxymethylfurfural and Phenolic Compounds in Honey

In the present work, a high-throughput field sample preparation method was reported for the simultaneous determination of 5-hydroxymethylfurfural and phenolic compounds in honey. Combining a simple and green homogenous liquid–liquid extraction, matrix-induced sugaring-out, with the use of a 96-deepwell plate and multichannel pipette, the proposed method showed its merits in instrument-free and high-throughput preparation. Due to the high-throughput property, the parameters of the method were rapidly and systematically studied using a constructed 4 × 2 × 4 × 3 array (sample amount × ratio of ACN:H₂O × standing time × replicates) in a 96-deepwell plate. Analytical performance was fully validated, and the limits of detection and limits of quantification were in the range of 0.17–1.35 μg/g and 0.51–4.14 μg/g, respectively. Recoveries were between 83.98 and 117.11%, and all the precisions were <5%. Furthermore, the developed method was successfully applied in the outdoor preparation of commercial honey samples and the in-field preparation of raw honey samples in apiary. The current work presented a simple, rapid, and high-throughput method for the field sample preparation of honey and provides a valuable strategy for the design of field and on-site sample preparation.     

食品中农药残留分析的样品前处理技术进展*

本文综述了近年来食品中农药残留分析的样品前处理技术,重点对超临界流体萃取法在食品农药残留分析中的应用及其联用技术进行了评述;同时对固相微萃取、微波辅助萃取和凝胶渗透色谱法进行了总结。对食物中农药残留分析技术的发展方向进行了讨论。