Individual and Synergistic Effects of Heavy-Metal Ions on the Induction of Cyanobacterial Metallothionein Examined by High-Performance Liquid Chromatography/Inductively Coupled Plasma Mass Spectrometry

Takatera, Kikuo; Watanabe, Tadashi

doi:10.2116/analsci.9.19

Cited by 13 publications

(7 citation statements)

References 15 publications

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“…Their general formula is (Glu- Cys) n Gly where n = 2-11. 111,153,314,325 HPLC-ICP-MS was proposed as a convenient technique for the monitoring of Cd, Cu and Zn complexes in aqueous extracts of microorganisms and plants that had been submitted to metal stress. The general structure of phytochelatins is conservative in a wide variety of plants but some modifications may occur on the C-terminal amino acid leading to the replacement of glycine by b-alanine, serine or glutamic acid.…”

Section: Metal Complexes With Phytochelatins and Soluble Proteinsmentioning

confidence: 99%

“…Quantification of signals has usually been done using peak area calibration either by converting (by removing the column) the measurement system into the flow injection mode after completing the chromatographic run, 322,325 or by using a calibration graph if standards were available. Quantification of signals has usually been done using peak area calibration either by converting (by removing the column) the measurement system into the flow injection mode after completing the chromatographic run, 322,325 or by using a calibration graph if standards were available.…”

Section: Standardization and Accuracymentioning

confidence: 99%

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Bio-inorganic speciation analysis by hyphenated techniques

Szpunar

2000

Analyst

279

197

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International audienceThe state-of-the-art of species-selective analysis for trace metals and metalloids in biological materials by chromatographic and electrophoretic separation techniques with element selective detection is critically reviewed. The species of interest include organoarsenic, organoselenium and metal complexes with bioglands such as phytochelatins, metallothioneins, proteins and polysaccharides. The separation mechanisms discussed include size-exclusion, anion- and cation-exchange and reversed-phase HPLC and flatbed and capillary zone electrophoresis. Advantages and limitations of various element selective (e.g. AAS, ICP-AES and ICP-MS) and molecule specific (electrospray MS/MS) detection techniques used on- and off-line are discussed. The applications of coupled techniques to the analysis of biological materials are comprehensively reviewed in tabular form. Attention is paid to the sample preparation and sources of error in bioinorganic speciation analysis

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Section: Metal Complexes With Phytochelatins and Soluble Proteinsmentioning

confidence: 99%

Section: Standardization and Accuracymentioning

confidence: 99%

Bio-inorganic speciation analysis by hyphenated techniques

Szpunar

2000

Analyst

279

197

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“…Quantification of signal obtained is usually done by a peak area calibration either by changing the measurement system into the flow-injection mode after completing the chromatographic run [21,24] or using a calibration g raph if standards are available [6,17].…”

Section: The Coupling Of Size-exclusion Chromatography With Icp-msmentioning

confidence: 99%

“…Metallothioneins in mussels have widely been studied [6,17]. Individual and synergistic effects of Cu, Zn and Cd ions on the induction of metallothionein in cyanobacterium (the origin of trophic chains in aquatic systems) were invest i gated [24]. The in vivo selenium association with cyanobacterial metallothioneins following the coadministration of Zn and selenite or selenate was examined [16].…”

Section: Ecotoxicologicalmentioning

confidence: 99%

The coupling of size-exclusion HPLC with ICP-MS in bioinorganic analysis

Makarov¹,

Szpunar²

1998

Analusis

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T he detection of a metal compound in a sample is the prerequisite of any further action concerning its identification, characterisation and the role in biochemistry. The actual problem can be reduced to proving that the analytical signal detected by the spectrometer is due to the presence of a metal compound and not to that of a simple ion. This can be done by preceding the detection by a separation technique that would differentiate between the free metal or metalloid ion and the same element bound with a larger, usually macromolecular structure.The detection of the presence of metal complexes with biological macromolecules has commonly been realised by ultrafiltration using a filter with a small cut-off molecular mass, usually 500 − 5 000 Da. The concentration of the element of interest was determined in the initial sample and in the fi l t rat e, u s u a l ly by graphite furnace AAS; the metal retained was considered to be bound to a macromolecular species. A refinement of this technique by the successive u l t ra fi l t ration through membranes with molecular we i g h t cut-offs of 500, 5 000 and 30 000 Da has widely been used to study the distribution of metal-species as a function of the molecular weight [1][2][3][4]. The method allows only a ro u g h speciation and is time consuming.An improvement in terms of the resolution can be obtained by the use of size-exclusion chromatography which also seems to be less cumbersome and faster allowing the on-line detection to be used. Because the resolution of SEC is insufficient for the discrimination of the small aminoacid heterogeneities, the coupling SEC-ICP-MS is the most popular technique for the first screening of an unknown sample in view of the presence of macromolecular species of elements. SEC using a short guard column with ICP-MS detection allows the rapid quantification of the bound metal fraction by comparison of the chromatographic signal with that, obtained in parallel, by flow-injection ICP-MS analysis of the sample. This paper presents the technique, reviews its applications and shows some recently established potential in the fields of the plant, animal and clinical biochemistry. Size-exclusion chromatography in bioinorganic analysisSize-exclusion chromatography (SEC) is based on the molecular sieve effect and enables species to be sep a rat e d according to their size, and to a lesser extent, shape. The average time a substance spends in the pores is determined by its size which for a given shape, can usually be related directly to its molecular weight. The following conditions have to be taken into consideration for separation of biological constituents containing metals: interactions between metals and bu ffe rs and ch ro m at ographic support mat e ri a l should be minimized, and the pH of the eluent should be weakly alkaline to prevent the metals from dissociating. PackingSeparation by SEC should be independent of the analyte's charge but in practice the stationary phase surface displays ch a rged pro p e rties so that a mixed mode sep a ra...

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“…Liquid chromatography followed by metal-specific detection by ICP-MS was widely applied to the detection and determination of MT and its isoforms in animal tissues. [8][9][10][11][12][13][14][15][16] The high separation efficiency of CZE for metalbinding peptides [17][18][19] has made many researchers choose rabbit liver metallothionein (MT), preparation available from Sigma-Aldrich, as the favourite test sample to demonstrate the performance of newly developed CZE-ICP-MS interface designs. [20][21][22][23] The recombinant mouse liver MT-1 was recently used for a study of metal complexation equilibria by CZE-ICP-MS. 24 However, no applications of CZE-ICP-MS to the characterization of metal complexes in animal tissues cytosols have been developed so far.…”

Section: Introductionmentioning

confidence: 99%