Detection and identification of rabbit liver metallothionein-2 subisoforms by capillary zone electrophoresis - inductively coupled plasma mass spectrometry and microbore HPLC - electrospray mass spectrometry

Chassaigne, Hubert; Mounicou, Sandra; Casiot, Corinne; Łobiński, Ryszard; Potin-Gautier, M.

doi:10.1051/analusis:2000280357

Cited by 14 publications

(9 citation statements)

References 11 publications

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“…Para o acoplamento com o ICP, a vazão empregada foi de 20 µL min -1 . Posteriormente, Chassaigne et al 54 também avaliaram uma interface empregando um nebulizador micro-concêntrico para a caracterização de metalotioneínas em solução de referência. Nesse caso, as características analíticas do sistema proposto também não foram apresentadas.…”

Section: Interface Ce-icp: Aplicações Em Química Analíticaunclassified

Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação

et al. 2003

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Recebido em 26/11/01; aceito em 25/7/02 CAPILLARY ELECTROPHORESIS COUPLED TO PLASMA SPECTROMETRY: AN EFFICIENT TOOL FOR SPECIATION. The most important features of the CE-ICP hyphenation, as well as its advantages and drawbacks as a tool for speciation are discussed. The fundamental principles of capillary electrophoresis and inductively coupled plasma mass spectrometry are also presented. Some applications involving different designs proposed in the literature to couple CE and ICP system for elemental speciation are reviewed.Keywords: speciation; CE-ICP-MS; CE-ICP-OES. INTRODUÇÃOOs metais pesados, largamente utilizados pela indústria, normalmente, não se encontram como íons livres ou em sua forma original no ambiente mas, ocorrem como parte de espécies químicas que são resultantes de uma série de processos naturais bio-geo-químicos, associados ou não às interferências humanas 1 . Isso significa que as diversas espécies de um mesmo elemento apresentam comportamentos diferenciados, o que pode comprometer a exatidão do método durante sua determinação 2 . Percebe-se, portanto, que é necessário fazer, além da determinação quantitativa total dos elementos presentes nas diferentes amostras, a identificação de cada espécie.As principais áreas de interesse da especiação envolvem o monitoramento da poluição nos solos, ar, águas, sedimentos e biota por espécies organometálicas e inorgânicas tóxicas. Muitos estudos relacionados à biometilação de metais (especialmente mercúrio) e semi-metais (arsênio e selênio) têm sido feitos, uma vez que estes elementos podem apresentar um alto nível de toxidez, dependendo do estado de oxidação em que se encontram 3 . Além disso, deve ser considerado que o estudo da cinética de elementos traços no corpo humano, devido à exposição aos metais, não seria possível sem as informações fornecidas pela especiação.O controle homeostático e metabolização de elementos essenciais, como zinco e cobre, e dos tóxicos, como mercúrio e arsênio, por organismos da biota também têm sido objeto de muitos estudos devido à importância econômica e às implicações ambientais da utilização desses elementos. Uma das áreas de maior interesse, no campo da especiação, é aquela relacionada às metalotioneínas (MT), as quais são um grupo de proteínas, não-enzimáticas, de baixa massa molar, ricas em cisteínas, resistentes à termo-coagulação e à precipitação ácida e que possuem um ligante metálico em sua estrutura. A principal característica das metalotioneínas é formar complexos com os metais bivalentes e complexos metal-tiol. Decorrente dessa característica, em sistemas biológicos, a síntese da proteína só ocorre induzida por metais 4 . Devido à importância das MT em sistemas biológicos, a escolha e otimização de sistemas acoplados para a determinação dos diferentes complexos com os metais ainda é motivo de vários estudos. Uma vez que as MT, para os diferentes organismos, possuem diferentes isoformas, a identificação e determinação de uma isoforma podem contribuir para uma melhor compreensão do papel e funções das isoforma...

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Section: Interface Ce-icp: Aplicações Em Química Analíticaunclassified

Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação

et al. 2003

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“…Investigations into the optimal configuration for ICP interfacing to CE, being a main trend during the past period, provided the method's practitioners with several effective custom-designed interfaces. Those that have proven most successful are the sheath-flow interface [61,63,64,66,[68][69][70][71][72][73][74][75][76][77][78][79][80][81][82][83][84] and the liquid-junction interface [65]. Whereas still assembled in-house but based on commercial equipment (a microconcentric nebulizer [63, 69-73, 76, 77, 81], Meinhard concentric nebulizer [61,74,75], a cross-flow nebulizer [65,72], a self-aspirating total consumption micronebulizer [79,80], a direct injection nebulizer [66], and other microflow nebulizers), these interfaces afford stable electrical connection and nebulizer operation, controlling laminar flow induced by nebulizer suction effect and, as a result, reproducible CE-ICP-MS performance.…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…This mode of MS detection appears useful for adding an extra selectivity dimension to CE data and particularly for obtaining structural information in order to facilitate the identification of (unknown) analytes. This attracting feature of ESI-MS may compensate for poorer detection limits compared to ICP-MS (also due to very low tolerance of nonvolatile electrolytes) and make CE-ESI-MS widely practiced for structural characterization, in the first instance, for elemental speciation purposes [20,67,69,71,73,81,90], including applications related to the field of pharmacology [91][92][93].…”

Section: Mass Spectrometrymentioning

confidence: 99%

“…Among other metal-bound biologically relevant compounds, which are in the focus of current CE speciesselective research, are MT, small cysteine-rich metal binding proteins. Naturally occurring as a mixture of basically two major differently charged isoforms with varying contents of Cd, Zn, and Cu, MTcan be well resolved using either uncoated fused-silica capillaries [69,71,73,78,79,81,82,251,252], or polymer-coated capillaries, neural [253,254] and negative [83], or polytetrafluoroethylene capillaries [116] and characterized by means of diode array [251,255], ICP-AES [116] or ICP-MS, utilized alone [69,79,89] or in parallel with ESI-MS [71,73,81]. The quantification of MT isoforms after the CE separation can be performed by use of ICP-sector field MS in combination with on-line isotope dilution [78,82,83].…”

Section: Speciation Analysismentioning

confidence: 99%

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Recent advances and trends in capillary electrophoresis of inorganic ions

Timerbaev

2002

Electrophoresis

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Capillary electrophoresis (CE) continues its invasion into the domain of inorganic analysis. This review summarizes the progress and significant developments of CE of inorganic ions over the first two years of the new century. As manifested by the substantial number of published articles, this field is still witnessing a growing activity of many separation scientists. To provide an in-depth treatment of fundamental and methodological aspects of inorganic ion analysis by CE, an extensive fraction of the relevant literature is covered whereas only most innovative application reports are briefly considered. Most of the recent advancements have focused on CE system modernization designed to improve a limited detection sensitivity, which still remains a major obstacle to overcome in order to ensure the method's wider practical acceptance. Attention is also given to how and in which directions the future research could be performed to implement CE in routine inorganic analysis.

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“…42 In this context, Mounicou et al 43 have compared CE-ICP-MS and CE-ES-MS systems for the characterization of MT reference solutions. Chassaigne et al 44 have also evaluated an interface employing a micro-concentric nebulizer for the characterization of MT in reference solutions. In the interface described by Prange et al, 45 an MCN from CETAC was used.…”

mentioning

confidence: 99%

Identification of a Metallothionein in Synechococcus by Capillary Electrophoresis Hyphenated with Inductively Coupled Plasma Mass Spectrometry

et al. 2003

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In 1957, Margoshes and Vallée 1 discovered a new type of protein, which seemed to contain metal ions in its structure. However, the first paper confirming the presence of cadmium in an isolated protein, named metallothionein (MT), was only published in 1960 by Kagi and Vallée. 2 Metallothioneins are proteins with low molecular mass, rich in cysteine residues and extremely reactive to bivalent metals. 3 These proteins are responsible for processes related to both intracellular absorption and the regulation of biologically essential elements, such as zinc and copper. MTs are also responsible for cell protection from toxic elements, such as cadmium and mercury. 4,5 Metallothioneins are present in mammals and also in eukaryote and prokaryote organisms. 3,6 In the majority of animal species, MTs are abundant in the liver, pancreas, intestines and kidney tissues. The concentration of these proteins depends on both the species and the tissues, and reveals the diet effect in each organism. 7 Metallothioneins are usually determined by employing classical methodologies, such as metal-ligand assays with mercury 8 and cadmium. 9 Among those classical methodologies are also found electrochemical techniques based on the quantification of MT fractions via the oxidation of sulfhydryl groups, 10 and also immunology techniques, such as radioimmunoassay 11 (RIA), and enzymatic assays in sorbents 12 (ELISA). However, methods involving metal saturation or electrochemical techniques undertake interferences due to oxyreduction reactions of other species present in the samples, 12 what makes them inefficient to quantify the metal concentration in the isoforms. 13 This drawback was overcome when anionexchange high-performance liquid chromatography (HPLC) was employed in the separation of the two main MT isoforms. 14 Subsequently, reverse-phase HPLC was also employed to improve the resolution in the separation of isoforms.15,16 HPLC was coupled to atomic absorption spectrometry (AAS) by using a T-shaped interface to determine Cd, Zn and Cu in MT from rabbit liver in µg g -1 level. 17,18 Other couplings, such as RP-HPLC with inductively coupled plasma mass spectrometry 19, and RP-HPLC with electrospray mass spectrometry 21,22 (ES-MS), were also proposed for the separation and determination of Cd and Zn in MT. Nevertheless, certain demands related to the utilization of organic solvents and column specificities restrict the use of RP-HPLC as a separation method. 23 In the last decade, capillary-zone electrophoresis has been successfully employed in the separation of proteins and peptides, especially for those with low molecular mass. 24,25 Despite its high efficiency, capillary electrophoresis (CE) presents some limitations related to the low sensitivity of UV detectors, which are widely employed in CE. 26 In this sense, it is not unexpected that several efforts have been made to couple CE with ICP-MS in order to overcome sensitivity limitations. However, there are some practical aspects, such as capillary positioning in the interface, flo...

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Detection and identification of rabbit liver metallothionein-2 subisoforms by capillary zone electrophoresis - inductively coupled plasma mass spectrometry and microbore HPLC - electrospray mass spectrometry

Cited by 14 publications

References 11 publications

Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação

Eletroforese capilar acoplada à espectrometria com plasma: uma ferramenta eficiente para a especiação

Recent advances and trends in capillary electrophoresis of inorganic ions

Identification of a Metallothionein in Synechococcus by Capillary Electrophoresis Hyphenated with Inductively Coupled Plasma Mass Spectrometry

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