Capabilities of fast protein liquid chromatography coupled to a double focusing inductively coupled plasma mass spectrometer for trace metal speciation in human serum

Bayón, Marı́a Montes; Soldado, A.; González, Elisa Blanco; Alonso, J. Ignacio García; Sanz‐Medel, Alfredo

doi:10.1039/a809026b

Cited by 37 publications

(28 citation statements)

References 24 publications

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“…However, the potential interaction of metal-protein complexes with the stationary and mobile phases of the chromatographic system could result in the poor recovery of the target analyte [14,17]. It is important to choose mild separation chromatographic methods to avoid artefacts in the resulting chromatograms.…”

Section: Lc-icp-ms Methods Developmentmentioning

confidence: 99%

“…It is known that SEC has the ability to separate biomolecules into ranges of different molecular weights, and ion exchange in the anion-exchange mode also offers an interesting alternative for the separation of metal biocompounds [12,13]. The use of anion exchange fast protein liquid chromatography (AE-FPLC) coupled with ICP-MS has provided satisfactory results when studying protein binding to metals such as aluminum, iron and vanadium [16,17].…”

Section: Introductionmentioning

confidence: 99%

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Study of tungstate–protein interaction in human serum by LC–ICP-MS and MALDI-TOF

2007

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Oral administration of sodium tungstate is an effective treatment for type 1 and 2 diabetes in animal models; it does not incur significant side effects, and it may constitute an alternative to insulin. However, the mechanism by which tungstate exerts its observed metabolic effects in vivo is still not completely understood. In this work, serum-containing proteins which bind tungstate have been characterized. Size exclusion chromatography (SEC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) with a Phenomenex Bio-Sep-S 2000 column and 20 mM HEPES and 150 mM NaCl at pH 7.4 as the mobile phase was chosen as the most appropriate methodology to screen for tungsten-protein complexes. When human serum was incubated with tungstate, three analytical peaks were observed, one related to tungstate-albumin binding, one to free tungstate, and one to an unknown protein binding (MW higher than 300 kDa). Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometric analysis of the tungsten-containing fractions collected from SEC-ICP-MS chromatograms, after desalting and preconcentration processes, confirmed the association of tungstate with albumin and the other unknown protein.

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Section: Lc-icp-ms Methods Developmentmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Study of tungstate–protein interaction in human serum by LC–ICP-MS and MALDI-TOF

2007

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“…FPLC has proved to be suitable for studying speciation of metal-proteins in biological fluids [12][13][14][15]. However, this column has been scarcely used for speciation studies of metallothioneins [21], in spite of the fact that it provides narrow sharp-pointed peaks in shorter time and better resolution than conventional anion-exchange columns.…”

Section: Discussionmentioning

confidence: 99%

“…However, such methodology has shown to provide poor resolution and reproducibility and rather long separation time. Therefore, the main objective of the present study has been to investigate the potential use of Fast Protein Liquid Chromatography (FPLC) on a Mono Q HR 5/5, previously used for trace element speciation of proteins in biological material [12][13][14][15], for the separation of such cadmium binding proteins. …”

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confidence: 99%

Improved separation of rabbit liver metallothioneins by FPLC-ICP-MS: a comparison with the conventional anion-exchange chromatography

Ferrarello¹,

Campa²,

Infante³

et al. 2000

Analusis

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Over the last decade, it has been documented that cadmium exposure can cause several health disorders including nephrotoxicity, hypertension, osteomalacia, lung disease and disturbed calcium metabolism [1,2]. Moreover, the International Agency for Research on Cancer (IARC) considers that cadmium and cadmium compounds are potentially carcinogenic to human (Group 1) [3,4].Metallothioneins (MT) play an important role for heavy metal detoxification, zinc and copper homeostasis, protection against oxidative damage caused by free radicals and as part of the acute phase response to inflammation and stress.The isoforms of MTs have structural similarity with the same number of cysteine residues and high metal binding affinity but differ in their total charge because of differences in certain amino acids other than cysteine.The polymorphism occurs during the evolution of the species and consists of the variation of the primary structure by the substitution of 1 to 15 amino acids. Hence, isoforms differ in amino acid composition and consequently, have different isoelectric points and different hydrophobicities.Isoforms with minor differences, such as one amino acid residue, were detected as subgroups of the major isoforms and are termed sub-isoforms [5]. It has been demonstrated that the different isoforms may be regulated by different inducers [6]. Experiments performed on cultured rabbit kidney cell [7] and human cell lines [8] demonstrated that various MT isoforms can be synthesised by distinct inducing factors and may play different cellular roles.Thus, the development of analytical methods for reliable measurement of the different isoforms of MTs, within a reasonable time, is nowadays urgently needed for many studies requiring MT isoforms information. Similarly, information about binding of toxic metals, e.g. Cd, to such MTs isoforms is desirable. That is, cadmium speciation studies in biological materials should be carried out [9]. Probably the coupling a powerful separation technique (e.g. chromatographic techniques) with an element specific detector (atomic detector) is the most popular approach for speciation purposes. For unknown species, electrospray ionisation MS studies are opening new exciting possibilities for a more detailed characterisation of metallothioneins in terms of their molecular mass [10].The use of a quadrupole ICP-MS detector coupled on-line with liquid chromatography is one of the most powerful modern tools for studying protein binding biometals in biological fluids. Thus, in our laboratory the determination of Cd-MT in human urine has been carried out by using a Protein Pak DEAE 5PW anion-exchange column coupled to ICP-MS via a vesicular hydride generation interface [11]. This hybrid technique was assessed for the cadmium speciation in standards of rabbit liver MT. However, such methodology has shown to provide poor resolution and reproducibility and rather long separation time. Therefore, the main objective of the present study has been to investigate the potential use of Fast Protein Liquid ...

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“…From the analytical point of view, high performance liquid chromatography (HPLC) and capillary electrophoresis (CE) are the most widely used methods to separate proteins. A comprehensive review on determination of metal-binding proteins by CE has been recently delivered by Stutz et al [22] The detection of the metals has been carried out with electrothermal atomic absorption spectrometry (ETAAS) [23,24], inductively coupled plasma-atomic emission spectrometry (ICP-AES) [25], and inductively coupled plasma-mass spectrometry (ICP-MS) that has been applied in the speciation of metals in human serum [26]. A complete review has been published on bio-inorganic speciation analysis by hyphenated techniques [27].…”

Section: Ironmentioning

confidence: 99%

Identification, characterization and determination of metal-binding proteins by liquid chromatography. A review

Guntiñas

Bordin

Rodríguez

2002

Analytical and Bioanalytical Chemistry

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The use of liquid chromatography in the separation and determination of metal-binding proteins is reviewed. Advantages and drawbacks of different chromatographic techniques based on various principles: size exclusion, ion exchange (cationic and ionic), reversed phase and affinity, are presented and discussed. The topic "metal-binding proteins" is considered and presented from two different points of view. The first one regards metal speciation in biological samples (serum and blood). In metal speciation studies, the exact identity of the protein to which the metal is bound often remains unknown. The second point of view is that, frequently, the interest of analyzing metal-binding proteins is not related anymore to the metallic fraction of the protein, but to other chemical structures attached to the protein, such as carbohydrates, which indirectly determine how good the function of the protein is. In this review, special attention is paid to studies dealing with the glycosylation of transferrin, and with the glycated isoform of haemoglobin.

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Capabilities of fast protein liquid chromatography coupled to a double focusing inductively coupled plasma mass spectrometer for trace metal speciation in human serum

Cited by 37 publications

References 24 publications

Study of tungstate–protein interaction in human serum by LC–ICP-MS and MALDI-TOF

Study of tungstate–protein interaction in human serum by LC–ICP-MS and MALDI-TOF

Improved separation of rabbit liver metallothioneins by FPLC-ICP-MS: a comparison with the conventional anion-exchange chromatography

Identification, characterization and determination of metal-binding proteins by liquid chromatography. A review

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