共查询到20条相似文献,搜索用时 15 毫秒
1.
Accelerator Mass Spectrometry (AMS) is an ultra‐sensitive analytical method which has been instrumental in developing microdosing as a strategic tool in early drug development. Considerable data is available for AMS microdosing using typical pharmaceutical drugs with a molecular weight of a few hundred Daltons. The so‐called biopharmaceuticals such as proteins offer interesting possibilities as drug candidates; however, experimental data for protein microdosing and AMS is scarce. The analysis of proteins in conjunction with early drug development and microdosing is overviewed and three case studies are presented on the topic. In the first case study AMS experimental data is presented, for the measured concentration of orally administered recombinant insulin in the blood stream of laboratory rabbits. Case study 2 concerns minimum sample size requirements. AMS samples normally require about 1 mg of carbon (10 µL of blood) which makes AMS analysis unsuitable in some applications due to the limited availability of samples such as human biopsies or DNA from specific cells. Experimental results are presented where the sample size requirements have been reduced by about two orders of magnitude. The third case study concerns low concentration studies. It is generally accepted that protein pharmaceuticals may be potentially more hazardous than smaller molecules because of immunological reactions. Therefore, future first‐in‐man microdosing studies might require even lower exposure concentrations than is feasible today, in order to increase the safety margin. This issue is discussed based on the current available analytical capabilities. Copyright © 2010 John Wiley & Sons, Ltd. 相似文献
2.
Summary The advantages that accelerator mass spectrometry (AMS) provide for radiocarbon analysis, notably smaller sample sizes and shorter measurement times, also apply to the analysis of 129I. In this paper, the requirements for a mass spectrometry system for measuring extremely low concentrations of rare atoms are discussed and these requirements are illustrated using the details of the AMS analysis of 129I. As an example of an application of this AMS technology, a series of 129I measurements, used to identify isolated events in which radioactivity has been atmospherically transported into the Arctic, is described. Such investigations could not be carried out without the small sample size capability of AMS analysis. 相似文献
3.
Accelerator mass spectrometry (AMS) is a nuclear physics technique developed about twenty years ago, that uses the high energy (several MeV) of a tandem Van de Graaff accelerator to measure very small quantities of rare and long-lived isotopes. Elements that are of interest in biomedicine and environmental sciences can be measured, often to parts per quadrillion sensitivity, i.e. zeptomole to attomole levels (10(-21)-10(-18) mole) from milligram samples. This is several orders of magnitude lower than that achievable by conventional decay counting techniques, such as liquid scintillation counting (LSC). AMS was first applied to geochemical, climatological and archaeological areas, such as for radiocarbon dating (Shroud of Turin), but more recently this technology has been used for bioanalytical applications. In this sphere, most work has been conducted using aluminium, calcium and carbon isotopes. The latter is of special interest in drug metabolism studies, where a Phase 1 adsorption, distribution, metabolism and excretion (ADME) study can be conducted using only 10 nanoCurie (37 Bq or ca. 0.9 microSv) amounts or less of 14C-labelled drugs. In the UK, these amounts of radioactivity are below those necessary to request specific regulatory approval from the Department of Health's Administration of Radioactive Substances Advisory Committee (ARSAC), thus saving on valuable development time and resources. In addition, the disposal of these amounts is much less an environmental issue than that associated with microCurie quantities, which are currently used. Also, AMS should bring an opportunity to conduct "first into man" studies without the need for widespread use of animals. Centre for Biomedical Accelerator Mass Spectrometry (CBAMS) Ltd. is the first fully commercial company in the world to offer analytical services using AMS. With its high throughput and relatively low costs per sample analysis, AMS should be of great benefit to the pharmaceutical and biotechnology industries as well as other life science areas. 相似文献
4.
It has been found that small mutations of certain genes are the definitive origin of many heritable disorders and cancers with striking development of recent molecular biology. Such new findings have taken close-up of the importance of gene mutation assays based on the difference of DNA base sequences in diagnostic or medical field Capillary electrophoresis can be a good candidate for an ideal method on such gene analysis, because the methods can be performed with trace amount of samples, high resolution and shorter running time. We have established an effect of oligonucleotide, which was introduced onto capillary inner surface, on the recognition of an overall sequence of sample DNA fragments as an affinity ligand. 相似文献
5.
Beryllium-7, mainly measured via γ-spectrometry, is used as a (natural) radiotracer for education and science. For activities?<?0.1 Bq and samples containing also longer-lived 10Be, accelerator mass spectrometry (AMS) is the method-of-choice. We demonstrate that 7Be and 10Be can be quantified at the Dresden AMS facility on the same prepared BeO. Detection limits (7Be) are?~?0.6 mBq. Samples as small as tens of millilitres of rainwater can be chemically processed (after acidification) within a few hours without expensive and slow ion exchange. Isobar (7Li) suppression by chemistry and AMS is sufficient to guarantee for an ultrasensitive, cheap, and fast detection method for 7Be allowing high sample throughput. 相似文献
6.
Aerosol Mass Spectrometers (AMS) are powerful tools in the analysis of the chemical composition of airborne particles, particularly organic aerosols which are gaining increasing attention. However, the advantages of AMS in providing on-line data can be outweighed by the difficulties involved in its use in field measurements at multiple sites. In contrast to the on-line measurement by AMS, a method which involves sample collection on filters followed by subsequent analysis by AMS could significantly broaden the scope of AMS application. We report the application of such an approach to field studies at multiple sites. An AMS was deployed at 5 urban schools to determine the sources of the organic aerosols at the schools directly. PM 1 aerosols were also collected on filters at these and 20 other urban schools. The filters were extracted with water and the extract run through a nebulizer to generate the aerosols, which were analyzed by an AMS. The mass spectra from the samples collected on filters at the 5 schools were found to have excellent correlations with those obtained directly by AMS, with r2 ranging from 0.89 to 0.98. Filter recoveries varied between the schools from 40 to 115%, possibly indicating that this method provides qualitative rather than quantitative information. The stability of the organic aerosols on Teflon filters was demonstrated by analysing samples stored for up to two years. Application of the procedure to the remaining 20 schools showed that secondary organic aerosols were the main source of aerosols at the majority of the schools. Overall, this procedure provides accurate representation of the mass spectra of ambient organic aerosols and could facilitate rapid data acquisition at multiple sites where AMS could not be deployed for logistical reasons. 相似文献
7.
近年来,蛋白质组学技术在样品前处理、分离技术和质谱检测技术方面获得了快速发展,已经可以实现在几小时内对上万种蛋白的同时定性和定量分析。然而,目前的主流蛋白质组学技术仍无法满足极微量生物样品,尤其是单细胞样品的组学分析需求。毛细管电泳分离技术具有峰宽窄、柱效高、样品用量少等优势,是与高分辨质谱在线联用的理想选择之一。该文评述了集成化和在线样品前处理以及主流的纳升液相色谱-质谱联用系统在高灵敏度蛋白质组学分析领域的发展现状和挑战,认为该领域的重要技术挑战之一在于目前的纳升液相色谱分离已经无法完全匹配现代高分辨质谱超过40 Hz的超高扫描速度,从而导致质谱使用效率的降低。针对上述技术挑战,该文重点探讨了毛细管电泳-质谱联用技术的独特技术优势和潜在发展机遇,主要包括:(1)面向微量酶解多肽样品的高柱效毛细管电泳分离。通过采用毛细管电色谱可以进一步改善毛细管电泳柱容量不足的局限;(2)面向高灵敏度分析的无鞘液/鞘液接口开发;(3)高效毛细管电泳分离与高扫描速度质谱检测的协同化使用。总之,我们预期毛细管电泳-质谱联用技术的进一步发展有望在针对单细胞等超微量生物学样品的蛋白质组学分析中获得更广泛的应... 相似文献
8.
DNA analysis plays a great role in genetic and medical research, and clinical diagnosis of inherited diseases and particular cancers. Development of new methods for high throughput DNA analysis is necessitated with incoming of post human genome era. A new powerful analytical technology, called microchip capillary electrophoresis (MCE), can be integrated with some experimental units and is characterized by high-speed, small sample and reagent requirements and high-throughput. This new technology, which has been applied successfully to the separation of DNA fragments, analysis of polymerase chain reaction (PCR) products, DNA sequencing, and mutation detection, for example, will become an attractive alternative to conventional methods such as slab gel electrophoresis, Southern blotting and Northern blotting for DNA analysis. This review is focused on some basic issues about DNA analysis by MCE, such as fabrication methods for microchips, detection system and separation schemes, and several key applications are summarized. 相似文献
9.
Ultrafast, membrane-mediated restriction digestion of DNA molecules followed by rapid gel microchip electrophoresis of the resulting fragments is described. Combination of restriction endonuclease digestion on small pore-size microfibrous membranes with sample loading and electrophoresis analysis in a multilane (up to 96) format resulted in very fast restriction digest based microscale DNA analysis. Complete digestion of several nanogram target DNA was accomplished on the microporous membrane at room temperature just in a few minutes with a single or a combination of various restriction enzymes, using only submicroliter quantities of samples and reagents. The reaction mixture containing membrane also served as sample loading device for the subsequent gel microchip electrophoresis based analysis. This work establishes methods for high-speed, high-throughput DNA analysis, featuring extremely low sample and reagent consumption, and fast restriction digestion in combination with sample loading and rapid gel microchip analysis of the resulting fragments. The entire restriction digestion, sample loading and electrophoresis analysis process required less than 20 min. 相似文献
10.
Standard three‐dimensional Fourier transform (FT) NMR experiments of molecular systems often involve prolonged measurement times due to extensive sampling required along the indirect time domains to obtain adequate spectral resolution. In recent years, a wealth of alternative sampling methods has been proposed to ease this bottleneck. However, due to their algorithmic complexity, for a given sample and experiment it is often hard to determine the minimal sampling requirement, and hence the maximal achievable experimental speed up. Herein we introduce an absolute minimal sampling (AMS) method that can be applied to common 3D NMR experiments. We show for the proteins ubiquitin and arginine kinase that for widely used experiments, such as 3D HNCO, accurate carbon frequencies can be obtained with a single time increment, while for others, such as 3D HN(CA)CO, all relevant information is obtained with as few as 6 increments amounting to a speed up of a factor 7–50. 相似文献
11.
In recent years there has been much progress in the miniaturization of sample treatment approaches for the analysis of organic contaminants. Whilst much focus has been given to analysis of liquid matrices (e.g., water, biological fluids), equivalent approaches for lipid rich biota samples have seen significantly less progress. This is especially the case for samples of very small organisms commonly employed as standard test species in ecotoxicity studies. Typically, the extractable biotic sample size available for body residue analysis is very small and the total pollutant accumulation can vary significantly between species types according to factors such as organism size, lipid content and exposure conditions. Depending on the physical and chemical characteristics of the analyte(s) in question, extraction and purification, especially from more complex matrices, appears to be one of the main bottlenecks in achieving their quantification. The current article presents a review of the available micro-extraction methods for small marine biota samples, focusing on environmentally important organic pollutants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and pesticides. 相似文献
12.
Measurement of the length of DNA fragments plays a pivotal role in genetic mapping, disease diagnostics, human identification and forensic applications. PCR followed by electrophoresis is used for DNA length measurement of STRs, a process that requires labeled primers and allelic ladders as standards to avoid machine error. Sequencing‐based approaches can be used for STR analysis to eliminate the requirement of labeled primers and allelic ladder. However, the limiting factor with this approach is unsynchronized polymerization in heterozygous sample analysis, in which alleles with different lengths can lead to imbalanced heterozygote peak height ratios. We have developed a rapid DNA length measurement method using peptide nucleic acid and dideoxy dNTPs to “tailor” DNA templates for accurate sequencing to overcome this hurdle. We also devised an accelerated “dyad” pyrosequencing strategy, such that the combined approach can be used as a faster, more accurate alternative to de novo sequencing. Dyad sequencing interrogates two bases at a time by allowing the polymerase to incorporate two nucleotides to DNA template, cutting the analysis time in half. In addition, for the first time, we show the effect of peptide nucleic acid as a blocking probe to stop polymerization, which is essential to analyze the heterozygous samples by sequencing. This approach provides a new platform for rapid and cost‐effective DNA length measurement for STRs and resequencing of small DNA fragments. 相似文献
13.
有机新污染物是一类在先进分析技术帮助下新鉴定的、现有法规未管制的、人为源的有机污染物.有机新污染物主要包括药品与个人护理、农药、全氟化合物、内分泌干扰物等,其会产生内分泌干扰效应、诱发抗性基因传播,还对人类和野生生物的生存与发展构成潜在威胁,因此检测环境样品中的有机新污染物浓度对生态环境和人体健康具有重大意义.由于环境... 相似文献
14.
以氨丙基甲基二乙氧基硅烷和正硅酸乙酯为硅源,三嵌段共聚物P123为模板剂,无机盐KCl为助剂,酸性条件下一步法合成了氨基-甲基双官能化SBA-15(AMS). XRD,FTIR,BET,29Si MAS NMR,SEM及HRTEM等表征结果表明:无机盐不仅提高了AMS材料的介孔有序度,而且控制样品形貌从六方平板状向纤维状转变. 胆红素吸附实验表明:六方平板状AMS比纤维状AMS和纯SBA-15具有更快的吸附速率和更大的吸附容量,这可能是六方平板状形貌易于胆红素扩散以及双官能基团(甲基,氨基)增加了对胆红素吸附作用力的结果. 相似文献
15.
Mass spectrometric methods for the trace analysis of inorganic materials with their ability to provide a very sensitive multielemental analysis have been established for the determination of trace and ultratrace elements in high-purity materials (metals, semiconductors and insulators), in different technical samples (e.g. alloys, pure chemicals, ceramics, thin films, ion-implanted semiconductors), in environmental samples (waters, soils, biological and medical materials) and geological samples. Whereas such techniques as spark source mass spectrometry (SSMS), laser ionization mass spectrometry (LIMS), laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), glow discharge mass spectrometry (GDMS), secondary ion mass spectrometry (SIMS) and inductively coupled plasma mass spectrometry (ICP-MS) have multielemental capability, other methods such as thermal ionization mass spectrometry (TIMS), accelerator mass spectrometry (AMS) and resonance ionization mass spectrometry (RIMS) have been used for sensitive mono- or oligoelemental ultratrace analysis (and precise determination of isotopic ratios) in solid samples. The limits of detection for chemical elements using these mass spectrometric techniques are in the low ng g −1 concentration range. The quantification of the analytical results of mass spectrometric methods is sometimes difficult due to a lack of matrix-fitted multielement standard reference materials (SRMs) for many solid samples. Therefore, owing to the simple quantification procedure of the aqueous solution, inductively coupled plasma mass spectrometry (ICP-MS) is being increasingly used for the characterization of solid samples after sample dissolution. ICP-MS is often combined with special sample introduction equipment (e.g. flow injection, hydride generation, high performance liquid chromatography (HPLC) or electrothermal vaporization) or an off-line matrix separation and enrichment of trace impurities (especially for characterization of high-purity materials and environmental samples) is used in order to improve the detection limits of trace elements. Furthermore, the determination of chemical elements in the trace and ultratrace concentration range is often difficult and can be disturbed through mass interferences of analyte ions by molecular ions at the same nominal mass. By applying double-focusing sector field mass spectrometry at the required mass resolution—by the mass spectrometric separation of molecular ions from the analyte ions—it is often possible to overcome these interference problems. Commercial instrumental equipment, the capability (detection limits, accuracy, precision) and the analytical application fields of mass spectrometric methods for the determination of trace and ultratrace elements and for surface analysis are discussed. 相似文献
16.
Accelerator mass spectrometry (AMS) is an ultra‐sensitive analytical method suitable for the detection of sub‐nM concentrations of labeled biological substances such as pharmaceutical drugs in body fluids. A limiting factor in extending the concentration measurements to the sub‐pM range is the natural 14C content in living tissues. This was circumvented by separating the labeled drug from the tissue matrix, using standard high‐performance liquid chromatography (HPLC) procedures. As the separated total drug amount is in the few fg range, it is not possible to use a standard AMS sample preparation method, where mg sizes are required. We have utilized a sensitive carbon carrier method where a 14C‐deficient compound is added to the HPLC fractions and the composite sample is prepared and analyzed by AMS. Using 50 µL human blood plasma aliquots, we have demonstrated concentration measurements below 20 fM, containing sub‐amol amounts of the labeled drug. The method has the immediate potential of operating in the sub‐fM region. Copyright © 2009 John Wiley & Sons, Ltd. 相似文献
17.
微萃取技术是分析化学领域发展迅速,且已经得到广泛应用的样品前处理技术.填充吸附剂微萃取(MEPS)是一种微量固相萃取技术,使用微量的吸附剂填充于微量注射器,通过反复抽推方式使样品多次流经吸附剂以完成样品吸附萃取过程,萃取后的样品可直接用于色谱分析.典型的MEPS萃取设备包括MEPS注射器和MEPS吸附床(BIN).ME... 相似文献
18.
Micro X-ray fluorescence (microXRF) has been used nondestructively to investigate elemental heterogeneity by constructing two-dimensional maps of elemental concentrations in reference materials. microXRF probes sample sizes well below the 100 mg mass usually recommended for reference materials by NIST. Multivariate methods of analysis, such as principal-component analysis (PCA), show promise in identifying whether "nugget" effects exist within a material, where an element is enriched in small, isolated areas of the sample. The PCA model is built based on data taken in one location and compared with each elemental map. This methodology is shown for several reference materials including SRM 2702 and SRM 2703 to show how PCA treatment can be used to identify which elements exhibit nugget effects within the sub-mg mass range. A method of calculating the minimum recommended mass for solid samples is suggested using PCA iteratively on X-ray maps from which adjacent data points have been averaged. This is repeated until the mass sampled in a map is indistinguishable from data taken at a single location, suggesting no nugget effects can be detected. For SRM 1577c, a mass as low as 370 microg can be used without measurable nugget effects. 相似文献
19.
A limitation of any current approach using solvent-free MALDI mass spectrometry is that only one sample at a time can be prepared and transferred to the MALDI-plate. For this reason, multiple-sample preparation approaches for solvent-free MALDI MS analysis of synthetic polymers were developed that are simple and practical. One approach multiplexed sample preparation by simultaneously preparing multiple samples. With this approach, as many as 384 samples could be prepared by addition of analyte, matrix, salt, and 1-mm metal beads to each well of a 384-well disposable bacti plate, capping the plate with the lid and homogenizing all samples simultaneously using a common laboratory vortex device. Besides the time savings achieved by a single vortex step for multiple samples, an additional advantage of this method relative to previously reported solvent-free preparation methods is that the mixing volume per sample is reduced, which allows a reduction in the amount of analyte required. This method, however, still requires the transfer of each homogenized sample to the MALDI plate for subsequent analysis. Here we report a novel approach that combines multiple simultaneous solvent-free sample preparation with automatic sample transfer to the MALDI target plate. This approach reduces the possibility of cross-contamination, the amount of sample and matrix consumed for an analysis, and the time required for preparation of multiple samples. These methods were shown to provide high-quality mass spectra for various synthetic polymer standards with M(n) values to 10 kDa. The methods are efficient in that small sample amounts are required, the sample/salt/matrix ratio is not critical, and the time necessary to achieve sufficient homogenization of multiple samples is less than 5 min. 相似文献
20.
Peptide fractionation is extremely important for the comprehensive analysis of complex protein mixtures. Although a few comparisons of the relative separation efficiencies of 2‐D methodologies using complex biological samples have appeared, a systematic evaluation was conducted in this study. Four different fractionation methods, namely strong‐cation exchange, hydrophilic interaction chromatography, alkaline‐RP and solution isoelectric focusing, which can be used prior to LC‐MS/MS analysis, were compared. Strong‐cation exchange × RPLC was used after desalting the sample; significantly more proteins were identified, compared with the nondesalted sample (1990 and 1375). We also found that the use of a combination of analytical methods resulted in a dramatic increase in the number of unique peptides that could be identified, compared with only a small increase in protein levels. The increased number of distinct peptides that can be identified is especially beneficial, not only for unequivocally identifying proteins but also for proteomic studies involving posttranslational modifications and peptide‐based quantification approaches using stable isotope labeling. The identification and quantification of more peptides per protein provide valuable information that improves both the quantification of, and confidence of protein identification. 相似文献
|