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Andreas Prange

Publications and source records attributed to Andreas Prange.

7 recordsLinked to original sources

Determination of sulfur and selected trace elements in metallothionein-like proteins using capillary electrophoresis hyphenated to inductively coupled plasma mass spectrometry with an octopole reaction cell.

The determination of sulfur in biologically relevant samples such as metalloproteins is described. The analytical methodology used is based on robust on-line coupling between capillary electrophoresis (CE) and octopole reaction cell inductively-coupled plasma mass spectrometry (ORC-ICP-MS). Polyatomic ions that form in the plasma and interfere with the determination of S at mass 32 are minimised by addition of xenon to the collision cell. The method has been applied to the separation and simultaneous element-specific detection of sulfur, cadmium, copper, and zinc in commercially available metallothionein preparations (MT) and metallothionein-like proteins (MLP) extracted from liver samples of bream ( Abramis brama L.) caught in the river Elbe, Germany. Instrumental detection limits have been calculated according to the German standard procedure DIN 32645 for the determination of sulfur and some simultaneously measured trace elements in aqueous solution. For sulfur detection limits down to 1.3 microg L(-1) ((34)S) and 3.2 microg L(-1) ((32)S) were derived. For the other trace elements determined simultaneously detection limits ranging from 300 ng L(-1) ((58)Ni) to 500 ng L(-1) ((66)Zn, (55)Mn) were achieved. For quantification of sulfur and cadmium in a commercially available MT preparation under hyphenated conditions the use of external calibration is suggested. Finally, the need for proper sample-preparation technique will be discussed.

Animals↗

CZE-ICP-MS separation of metallothioneins in human brain cytosols: comparability of electropherograms obtained from different sample matrices.

Capillary zone electrophoresis, with its high resolution capability in the separation of different compounds, is well suited for the investigation of metal-containing proteins, especially when elemental detection is conducted using hyphenated inductively coupled plasma-mass spectrometry. A major problem in the separation of proteins in body fluids is caused by the effects of different sample matrix composition. The migration time of proteins varies significantly, depending on the nature of the matrix. Electropherograms are consequently difficult to compare and the peak identification is uncertain. Pre-analytical steps for the reduction of matrix compounds enhance the quality of the data, but the results are still unsatisfactory. This paper describes a technique for obtaining electropherograms that can be used for comparison purposes by correction of the data with the aid of time markers. A mixture of five substances (caesium chloride, arsenocholine, arsenobetaine, dimethylarsinic acid and monomethylarsonic acid) was added in a separate injection step. Ionic caesium eluted at the start of the separation and the other four markers appeared throughout and at the end of the electropherogram. All electropherograms were normalized to a reference run by recalculation of the time axis using the time markers. The method was applied to the analysis of human brain cytosols. Samples were separated after different pre-treatment steps and were compared, with special emphasis on the detection of the isoform metallothionein-3.

Brain Chemistry↗

Hyphenated techniques for the characterization and quantification of metallothionein isoforms.

Recent developments in the coupling of highly selective separation techniques such as capillary electrophoresis (CE) and high-performance liquid chromatography (HPLC) to element-specific and molecule-specific detectors, such as inductively-coupled plasma mass spectrometry (ICP-MS) and electrospray ionization-tandem mass spectrometry (ESI-MS/MS) for the characterization and quantification of metallothioneins (MTs) are critically reviewed and discussed. This review gives an update based on the literature over the last five years. The coupling of CE to ICP-MS is especially highlighted. As a result of progress in new interface technologies for CE-ICP-MS, research topics presented in the literature are changing from "the characterization of interfaces by metallothioneins" to the "characterization of metallothioneins by CE-ICP-MS". New applications of CE-ICP-MS to the analysis of MTs in real samples are summarized. The potential of the on-line isotope dilution technique for the quantification of MTs and for the determination of the stoichiometric composition of metalloprotein complexes is discussed. Furthermore, a selection of relevant papers dealing with HPLC-ICP-MS for MT analysis are summarized and compared to those dealing with CE-ICP-MS. In particular, the use of size-exclusion (SE)-HPLC as a preliminary separation step for metallothioneins in real samples prior to further chromatographic or electrophoretic separations is considered. Additionally, the application of electrospray ionisation-tandem mass spectrometry (ESI-MS/MS) for the identification of metallothionein isoforms following electrophoretic or chromatographic separation is discussed.

Animals↗

Use of surface-modified capillaries in the separation and characterization of metallothionein isoforms by capillary electrophoresis inductively coupled plasma mass spectrometry.

The separation of metallothionein (ML) isoforms using capillary electrophoresis (CE) has been improved by applying surface-modified capillaries, and the metal composition of MTs has been characterized by subsequent inductively coupled plasma sector field mass spectrometry (ICPSFMS). Nine MT complexes in a commercial preparation from rabbit liver were successfully separated on an anionic polymer-coated column, prepared by immobilizing poly(2-acrylamido-2-methyl-1-propanesulfonic acid) on the fused-silica surface via a linking agent. On uncoated capillaries or those coated dynamically with cationic materials, only three complexes could be separated. Online isotope dilution analysis combined with CE/ICPSFMS indicated the stoichiometric molar metal contents in the MT complexes.

Animals↗

Characterization and quantification of metallothionein isoforms by capillary electrophoresis-inductively coupled plasma-isotope-dilution mass spectrometry.

In a new approach to the characterization and quantification of metallothionein isoforms an on-line isotope-dilution method in combination with the coupling of capillary electrophoresis (CE) to an inductively coupled plasma-sector field mass spectrometer (ICP-SFMS) is reported. Metallothionein (MT) isoforms are separated by CE and the elements Cu, Zn, Cd, and S are detected simultaneously by use of ICP-SFMS in the medium resolution mode. On-line isotope dilution is performed by continuous introduction of an isotopically enriched, species-unspecific spike solution after the separation step. MT from rabbit liver and a further purified MT-1 isoform were quantified by determination of sulfur, and the stoichiometric compositions of the metalloprotein complexes are characterized by determination of their sulfur-to-metal ratios.

Algorithms↗