原子光谱/元素质谱在生命分析中的应用进展

原子光谱/元素质谱是元素分析的强有力手段,其在生命分析领域的应用也越来越广泛。在单细胞元素分析方面,相关研究工作主要关注元素在单细胞中的分布和形态变化;在元素标记策略分析领域,利用原子光谱（atomic spectrometry, AS）和电感耦合等离子体质谱（inductively coupled plasma mass spectrometry, ICP-MS）实现对小分子、核酸、蛋白质等目标分析物的高灵敏检测是研究热点;在金属药物分析领域,ICP-MS为研究金属药物在生物体中的摄入、分布、代谢和排泄等过程提供了便利,也为进一步阐明药物作用机理以及金属药物的设计和改进提供了数据支持;在生物元素成像领域,ICP-MS与激光剥蚀技术（laser ablation, LA）联用,可以对生物样品进行原位分析和微区分析,结合有机质谱实现元素相关生物过程的分子机制研究;与相关分离方法联用,原子光谱和元素质谱还可以对生物组织中元素进行形态分析,研究其在相关过程中的生物转化过程。本文从单细胞元素分析、元素标签标记策略、金属药物转运与代谢以及生物组织中元素分布分析等方面,评述了原子光谱和ICP-MS在生命分析中的应用实例,并对该领域的发展前景进行了展望。     

毛细管电泳-原子光(质)谱联用技术在金属形态与生物分子相互作用研究中的应用

金属形态与生物分子相互作用研究对于揭示金属元素在正常生命过程、重大疾病的发生、诊断和治疗过程中的作用机理具有重要意义,发展研究金属元素形态与生物活性分子相互作用的新技术和新方法非常重要。简要总结了十余年来,在发展毛细管电泳与电热原子吸收光谱和毛细管电泳与电感耦合等离子体质谱在线联用新技术,及其在不同形态金属与生物分子相互作用机理研究方面的工作。这些工作利用毛细管电泳-原子光(质)谱联用技术不仅能够直接证明镉、锌、不同形态的汞、不同形态的锑与谷胱甘肽、DNA、人血清白蛋白、牛血清白蛋白间的相互作用,以及这些金属及其不同形态与生物分子加合物的生成,而且还能够测定相互作用的热力学参数、反应级数和动力学参数。另外,结合圆二色光谱、红外光谱光谱、拉曼光谱和X射线光电子能谱等实验手段,进一步研究了金属及其不同形态与生物分子作用对生物分子二级结构的影响及在生物分子上可能的结合位点等。     

毛细管电泳-原子光(质)谱联用技术在金属形态与生物分子相互作用研究中的应用SCIEI北大核心CSCD

金属形态与生物分子相互作用研究对于揭示金属元素在正常生命过程、重大疾病的发生、诊断和治疗过程中的作用机理具有重要意义,发展研究金属元素形态与生物活性分子相互作用的新技术和新方法非常重要。简要总结了十余年来,在发展毛细管电泳与电热原子吸收光谱和毛细管电泳与电感耦合等离子体质谱在线联用新技术,及其在不同形态金属与生物分子相互作用机理研究方面的工作。这些工作利用毛细管电泳-原子光(质)谱联用技术不仅能够直接证明镉、锌、不同形态的汞、不同形态的锑与谷胱甘肽、DNA、人血清白蛋白、牛血清白蛋白间的相互作用,以及这些金属及其不同形态与生物分子加合物的生成,而且还能够测定相互作用的热力学参数、反应级数和动力学参数。另外,结合圆二色光谱、红外光谱光谱、拉曼光谱和X射线光电子能谱等实验手段,进一步研究了金属及其不同形态与生物分子作用对生物分子二级结构的影响及在生物分子上可能的结合位点等。     

ICP-MS在PM_(2.5)元素分析中的应用进展

大气细颗粒物(PM_(2.5))粒径小,比表面积大,容易吸附金属、有机物、病毒、细菌等污染物而成为有毒有害物质的载体和反应体,严重影响空气质量,现已成为当前大气环境的首要污染物,而其中金属及类金属由于具有非降解性和滞后性,严重污染自然环境,当PM_(2.5)被吸入人体内,有毒有害金属及类金属元素由呼吸道沉积在肺泡,而后转移至血液及其他器官中,可对人体正常生理机能产生影响,造成身长发育缓慢,甚至导致癌症等病变,进而严重威胁人体健康。近年来,我国许多城市也相应开展了PM_(2.5)中金属元素污染特征、分布水平及源解析的研究。选择有效采集PM_(2.5)中的金属及类金属元素的方法,消解效率较高的前处理方法以及操作简便、快速、准确、灵敏和抗干扰能力强的检测方法已成为当前PM_(2.5)中元素分析的研究重点和热点领域。而电感耦合等离子质谱(ICP-MS)法测定PM_(2.5)中金属及类金属元素,不仅能满足多元素同时测定,而且动态线性范围宽,检出限低,灵敏度高,国内外学者们已进行了大量的研究工作,形成了比较完善的研究体系。该分析方法可为PM_(2.5)中各金属及类金属组成及来源、时空分布、形态及相应同位素分析、生理毒性和转化机制等方面的研究工作提供强有力的数据支持。主要对ICP-MS测定PM_(2.5)中金属及类金属元素的分析方法进行了综述,着重对其采样滤膜选择、前处理方法及其消解液的选择进行了详述,重点阐述了ICP-MS联用技术在PM_(2.5)金属和类金属元素形态及同位素分析中的应用研究,总结了各种采样滤膜、前处理方法和消解液及检测联用技术各自的优缺点和选择依据,并对该领域未来存在的挑战和研究方向提出了展望,为进一步发展更简便、快速、高灵敏且选择性好的PM_(2.5)元素分析中ICP-MS检测技术提供参考。     

光谱分析方法在同位素检测中的研究和应用进展

同位素在核工业为主的各种工业生产中受到广泛的关注,并推动着地质学、材料科学、化学等相关学科的发展。近年来,基于光谱分析原理的同位素分析方法的开发逐渐受到关注。虽然多接收杯电感耦合等离子体质谱（MC-ICP-MS）、热电离质谱（TIMS）和气体同位素质谱（IRMS）等质谱技术是同位素分析的标准方法,但是这些质谱方法通常需要复杂的样品前处理流程以及频繁的仪器维护。光谱分析方法在这些方面有着自身独特优势,甚至可以满足现场实时快速的同位素分析,并在核工业同位素分析和传统稳定同位素分析领域已经取得了日益广泛的应用。随着光谱仪器关键部件和数据处理方法的进一步发展,极大地改善了光谱法同位素分析的性能（灵敏度、分辨率和精密度）,使光谱分析方法被逐渐开发并应用于环境和地质同位素分析领域。综述了光谱分析方法在同位素分析（定量或定性）领域的主要进展,从光谱分析原理的角度归类为发射光谱（原子发射、分子发射、拉曼光谱）和吸收光谱（原子吸收、分子吸收）两大类。着重讨论了光谱法进行同位素分析的基本原理、发展历程以及重要进展,简述了与质谱法相比的优缺点。针对仍然有待突破的技术难点,展望了光谱法应用于同位素分析的发展前景。该综述可为光谱分析方法在同位素检测中的发展方向提供重要参考。     

光腔衰荡光谱技术测定大气水汽稳定同位素校正方法研究

几乎所有小的气相分子（如H₂O,CO₂等）均具有独特的近红外吸收光谱,在负压条件下,每种微小的气相分子都拥有一对一的特征光谱线,基于这一原理人们开始使用激光光谱（IRIS）技术来准确分析气体样品中的同位素组成。该方法克服了传统同位素比质谱（isotope ratio mass spectrometry, IRMS）方法的局限性,已经成为公认的高精度、高灵敏度和高准确度的痕量气体检测方法。以大气水汽稳定同位素研究为例,大气水汽稳定同位素组成对水汽源区及其通道上的输送过程等水循环研究有着重要的指示意义。激光光谱技术使得大气水汽氢氧稳定同位素（δ18O和δD）野外原位连续高分辨率观测成为可能。但是,其观测精度和准确度受仪器运作特点、不同浓度大气水汽对特定光谱吸收性的敏感性差异等因素的影响,通常观测结果具有明显的非线性响应问题。因此,有必要对仪器观测过程中出现的各种偏差进行校正,但现阶段许多用户对新观测技术的国际校正方法尚不清楚。因此,基于波长扫描-光腔衰荡光谱（WS-CRDS）技术的大气水汽同位素观测系统（Picarro L2120-i）,通过可调谐二极管激光器（Tunable Diode Laser, TDL）发射可被待测气体分子所吸收的不同波长的激光,测量不同波长下的衰荡时间（即有样品吸收的衰荡时间）;TDL发射不能被待测气体吸收的不同波长的激光,测量每个波长下的衰荡时间（相当于无样品吸收的衰荡时间）。通过分析有无样品吸收的衰荡时间差,高精度计算待测气体的分子浓度,进而计算水汽稳定同位素组成。从记忆效应、漂移效应、浓度效应等方面,系统建立了一套准确可靠的大气水汽稳定同位素观测流程与校正方法,为正在使用或将要使用此类设备的研究人员提供参考,以获得高精度和高可靠性的大气水汽稳定同位素观测数据。     

质谱探微

质谱分析法是化学领域中非常重要的一种分析方法，将用于分析的样品分子(或原子)电离成具有不同质量的单电荷或碎片离子，进入由电场和磁场组成的分析器，然后用检测系统进行检测，即可得到不同质荷比的谱线即质谱．通过质谱分析，可以获得分析样品的分子量、分子式、分子中同位素构成和分子结构等多方面的信息．质谱分析法敏感性很强，可以用它追踪各种数量极少的分子．     

光质谱法探测不同状态下机动车尾气中芳香烃物质相对含量的变化

利用高灵敏度、高选择性、多组分和快速实时分析探测的方法——激光质谱方法研究机动车尾气。发现在266nm激光作用下,低速状态下机动车尾气中的芳香族含量远高于加速状态下的含量。     

基于上转换发光技术的生物传感器及其应用

为实现对特定生物分子的高灵敏度快速检测与分析。采用上转换发光材料作为标记物,研制成功一台基于上转换发光技术的新型光学免疫生物传感器。该传感器利用上转换发光材料在红外光激发下发射可见磷光的特性,通过对免疫层析试纸条上经生物反应而结合上去的上转换发光材料颗粒的含量进行检测,计算出被测样品中特定生物分子的浓度。实验结果表明,该传感器具有较好的生物特异性,对兔抗鼠疫免疫球蛋白(IgG)标准样品的检测灵敏度达到ng／ml量级,并在200～6000ng／ml浓度范围内具有良好的线性响应特性,相关系数R^2≥0．95;对鼠疫耶尔森氏菌抗体的敏感性明显高于间接血凝实验,且与免疫印迹检测实验结果具有较好的一致性。该传感器具有稳定、可靠、灵敏的工作性能,符合实际检测与分析的要求。     

金属电极对有机分子非弹性电子隧穿谱的影响

利用第一性原理计算金属电极下1,6-己二硫醇和1,4-二巯基苯分子结的非弹性电子隧穿谱,发现非弹性电子隧穿谱对金属电极的变化十分灵敏,并且非弹性电子隧穿谱的振动峰位置和强度与硫原子和金属电极表面的距离密切相关.结果表明电极材料和分子与金属成键的情况是影响分子结的非弹性电子输运的重要因素.理论分析进一步表明不同金属电极和有机分子的耦合能不同导致了谱峰强弱的调整.     

Atomic spectrometry and atomic mass spectrometry in bioanalytical chemistry

Abstract

Atomic spectrometry and atomic mass spectrometric (MS) techniques have been playing crucial roles in the field of biosciences. They detect elements with relatively high sensitivities and are thus applicable to a wide range of analytical targets. In the past decade, determination of bio-relevant metallic elements continues to be of interest, while particularly noteworthy are methods developed for small molecules, peptides, proteins, nucleic acids and even cells that well exploited the bio-analytical strengths of atomic spectrometry and atomic MS, either in a direct or indirect manner. Quantitation, as well as speciation and imaging analyses are all involved. The present review aims to assimilate recent advances in bio-analysis utilizing atomic spectrometry and atomic MS, primarily covering the period of 2013–2018, in an attempt to provide readers insight into the developing trends of this research frontier. Followed by concluding remarks and perspectives, the applications are divided into the following four catalogs: (i) toxicologically important metal-containing species, with an emphasis on quantitative and imaging analysis; (ii) quantitation of biomolecules using naturally occurring heteroatoms; (iii) exogenous metal ion or nanoparticle tagging-based strategies in bioassays; and (iv) label-free detection of biomolecules.     

Analytics/Methods

An overview is presented of biomedical applications of stable isotopes in general, but mainly focused on the activities of the Center for Liver, Digestive and Metabolic Diseases of the University Medical Center Groningen. The aims of metabolic studies in the areas of glucose, fat, cholesterol and protein metabolism are briefly explained, as well as the principle of breath testing and the techniques to study body composition and energy expenditure. Much attention is paid to the analytical considerations based upon metabolite concentrations, sample size restrictions, the availability of stable isotope labelled substrates and dose requirements in relation to compound-specific isotope analysis. The instrumental advantages and limitations of the generally used techniques gas chromatography/reaction/isotope ratio mass spectrometry and gas chromatography/mass spectrometry are described as well as the novelties of the recently commercialised liquid chromatography/combustion/isotope ratio mass spectrometry. The present use and future perspective of infrared (IR) spectrometry for clinical and biomedical stable isotope applications are reviewed. In this respect, the analytical demands on IR spectrometry are discussed to enable replacement of isotope ratio mass spectrometry by IR spectrometry, in particular, for the purpose of compound-specific isotope ratio analysis in biological matrices.     

Analytical techniques in biomedical stable isotope applications: (isotope ratio) mass spectrometry or infrared spectrometry?

An overview is presented of biomedical applications of stable isotopes in general, but mainly focused on the activities of the Center for Liver, Digestive and Metabolic Diseases of the University Medical Center Groningen. The aims of metabolic studies in the areas of glucose, fat, cholesterol and protein metabolism are briefly explained, as well as the principle of breath testing and the techniques to study body composition and energy expenditure. Much attention is paid to the analytical considerations based upon metabolite concentrations, sample size restrictions, the availability of stable isotope labelled substrates and dose requirements in relation to compound-specific isotope analysis. The instrumental advantages and limitations of the generally used techniques gas chromatography/reaction/isotope ratio mass spectrometry and gas chromatography/mass spectrometry are described as well as the novelties of the recently commercialised liquid chromatography/combustion/isotope ratio mass spectrometry. The present use and future perspective of infrared (IR) spectrometry for clinical and biomedical stable isotope applications are reviewed. In this respect, the analytical demands on IR spectrometry are discussed to enable replacement of isotope ratio mass spectrometry by IR spectrometry, in particular, for the purpose of compound-specific isotope ratio analysis in biological matrices.     

Detection of hydrogen isotope atoms by mass spectrometry combined with resonant ionization

We examined the application of mass spectrometric methods using resonant ionization by a tunable laser and proposed its use for analyzing hydrogen isotopes. We conducted resonance ionization mass spectrometry (RIMS) to detect gas-phase hydrogen isotope atoms. The ionization efficiency was increased by more than 1000 times that obtained with conventional methods using nonresonant ionization. Resonant laser ablation mass spectrometry (RLAMS) was applied for deuterium detection in solid samples. A graphite substrate implanted with deuterium was used for ordinary laser ablation mass spectrometry (LAMS) and RLAMS. The deuterium signal was observed very clearly by RLAMS, in contrast to LAMS. Mass spectrometry combined with resonance ionization was very useful for hydrogen isotope detection, because components with equal mass numbers were resolved and the method demonstrated higher ionization efficiency. Received: 4 November 1998 / Revised version: 12 January 1999 / Published online: 7 April 1999     

Thermal Ionization and Inductively Coupled Plasma Mass Spectrometry: Potential for Application to Studies on the Biokinetics of Molybdenum in Humans

Abstract

The use of stable isotopes as tracers in biokinetic investigations provides a means to obtain important metabolic data directly in humans without exposing the subjects to undue risks. In this work, three types of mass spectrometers are compared with regard to the determination of the abundances of stable isotopes of molybdenum in natural and enriched aqueous samples. The data show a good response of thermal ionization mass spectrometry (TIMS) and of high-resolution inductively coupled plasma mass spectrometry (ICPMS) to the isotopic enrichment of the samples, whereas conventional quadrupole ICPMS shows an unsatisfactory reproducibility of the results. Moreover, only TIMS can achieve an accuracy of better than 1% for the obtained isotopic ratios. Although a tedious procedure for the preparation of the biological samples is required and less sensitivity is achieved as compared to ICPMS, TIMS still seems to be method of choice for the accurate assessment of isotope ratios as required in multitracer studies on human biokinetics of trace metals.     

The role of technology in the past and future development of the doubly labelled water method

The doubly labelled water method is an isotope-based technique that is used to measure the energy demands of free-living animals and humans. It is based on the observation that, in the body, the oxygen in carbon dioxide is in complete isotope exchange equilibrium with the oxygen in body water. Hence, a label of isotopic oxygen in body water is eliminated by both respiratory CO(2) and water turnover, whereas a similarly introduced label of deuterium is eliminated only by water flux. The difference in isotope fluxes therefore permits estimation of CO(2) production, which is correlated to energy demands. The doubly labelled water method has been advanced predominantly by technological advances in mass spectrometry. Although it was first described in the 1950s, it was only used on small animals and in low numbers because the costs of the isotopes were a primary constraint. However, advances in mass spectrometry precision and accuracy in the 1980s made it possible to reduce the quantities of isotope used, and hence apply the method on humans, although still in small numbers. The advent of continuous flow inlets in the 1990s made possible the processing of samples in much larger numbers and the sample sizes of studies have expanded. Ironically, however, the technique is now under treat because of technological advances in another area (positron emission tomography), which has generated an enormous demand for (18)O and pushed up the price of isotopes. A continuation of this trend might drive prices to levels where sustained application of the method in human studies is questionable. Replacing determination of isotope enrichments currently performed by isotope ratio mass spectrometry with determinations made by stable isotope infrared laser spectrometry may be a technological advance that will get us out of this problem.     

Isotope shifts and hyperfine structure in calcium 4snp and 4snf F Rydberg states

Isotope shifts and hyperfine structure have been measured in 4snp ¹ P₁ and Rydberg states for all stable calcium isotopes and the radioisotope ⁴¹Ca using high-resolution laser spectroscopy. Triple-resonance excitation via Rydberg state was followed by photoionization with a CO₂ laser and mass selective ion detection. Isotope shifts for the even-mass isotopes have been analyzed to derive specific mass shift and field shift factors. The apparent isotope shifts for ⁴¹Ca and ⁴³Ca exhibit anomalous values that are n-dependent. This is interpreted in terms of hyperfine-induced fine-structure mixing, which becomes very pronounced when singlet-triplet fine-structure splitting is comparable to the hyperfine interaction energy. Measurements of fine-structure splittings for the predominant isotope ⁴⁰Ca have been used as input parameters for theoretical calculation of the perturbed hyperfine structure. Results obtained by diagonalizing the second-order hyperfine interaction matrices agree very well with experimentally observed spectra. These measurements allow the evaluation of highly selective and sensitive methods for the detection of the rare ⁴¹Ca isotope. Received 17 December 1999 and Received in final form 29 March 2000     

Laser resonance ionization for ultra-trace analysis on long-lived radioactive isotopes

Benefiting from the continuous laser developments, resonance ionization can be applied for a variety of experiments on radioactive isotopes, e.g. as a laser ion source for producing pure beams of short-lived isotopes at on-line facilities. In this paper the application of a compact set-up for resonance ionization mass spectrometry for ultra-trace analysis of the long-lived isotope Ca-41 is described. With this set-up a purely optical selectivity of 3×10⁹ and an overall detection efficiency of 1.2(4)×10⁻⁵ are demonstrated.     

Evaluation of the performance of high temperature conversion reactors for compound-specific oxygen stable isotope analysis

In this study conversion conditions for oxygen gas chromatography high temperature conversion (HTC) isotope ratio mass spectrometry (IRMS) are characterised using qualitative mass spectrometry (IonTrap). It is shown that physical and chemical properties of a given reactor design impact HTC and thus the ability to accurately measure oxygen isotope ratios. Commercially available and custom-built tube-in-tube reactors were used to elucidate (i) by-product formation (carbon dioxide, water, small organic molecules), (ii) 2nd sources of oxygen (leakage, metal oxides, ceramic material), and (iii) required reactor conditions (conditioning, reduction, stability). The suitability of the available HTC approach for compound-specific isotope analysis of oxygen in volatile organic molecules like methyl tert-butyl ether is assessed. Main problems impeding accurate analysis are non-quantitative HTC and significant carbon dioxide by-product formation. An evaluation strategy combining mass spectrometric analysis of HTC products and IRMS ¹⁸O/¹⁶O monitoring for future method development is proposed.     

Measuring technique for thermal ionisation mass spectrometry of human tracer kinetic study with stable cerium isotopes

Thermal ionisation mass spectrometry (TIMS) method has been developed for the simultaneous detection of different cerium isotopes in biological samples (i.e., blood and urine) at very low concentrations. The work has been done in the frame of a biokinetic study, where different stable cerium isotopes have been administered orally and intravenously as tracers to the human body. In order to develop an appropriate detection method for the tracers in the biological samples, an optimum sample preparation technique has been set and adapted to the specific requirements of the analysis technique used, i.e., TIMS. For sample evaporation and ionisation, the double tantalum filament technique showed the best results. The ions produced were simultaneously collected on a secondary electron multiplier so that the isotopic ratios of the cerium isotopes in the biological samples could be measured. The technique has been optimised for the determination of cerium down to 1?ng loaded on the evaporation filament corresponding to cerium concentrations of down to 1?ng?ml(-1) in the blood or urine samples. It has been shown that the technique is reliable in application and enables studies on cerium metabolism and biokinetics in humans without employing radioactive tracers.