Development of Miniaturized HPLC-ICP-MS for Speciation of Bio-Trace Elements
Abstract
References40
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Narrow-bore HPLC-ICP-MS for speciation of copper in mutant mouse neonates bearing a defect in Cu metabolism
Minute amounts of tissue supernatants from mouse neonates bearing a mutation in the copper (Cu)-transporter gene, Atp7a, were injected into narrow-bore HPLC coupled with an inductively coupled plasma–mass spectrometer (ICP–MS) to examine Cu metabolism. In the 14-day-old mutant neonates, Cu accumulated in the intestine in the metallothionein (MT)-bound form, and mRNA expression of the two MT isoforms was increased. Meanwhile, Cu in the MT-bound form (Cu-MT) was depleted in the liver and mRNA expr...
Separation of metallothionein isoforms extracted from isoform-specific knockdown cells on two-dimensional micro high-performance liquid chromatography hyphenated with inductively coupled plasma-mass spectrometry
A two-dimensional (2D) micro HPLC-inductively coupled plasma mass spectrometry (ICP-MS) system consisting of a gel filtration column and an anion-exchange column was constructed to separate two metallothionein (MT) isoforms. MT-I and MT-II genes were specifically knocked down with the RNA interference (RNAi) technique, and the effects of the knockdown were evaluated by the reverse transcription-polymerase chain reaction (RT-PCR) and the 2D micro HPLC-ICP-MS system. The minimum cell number requir...
Selenopeptide mapping in a selenium-yeast protein digest by parallel nanoHPLC-ICP-MS and nanoHPLC-electrospray-MS/MS after on-line preconcentration
ICP collision cell MS was optimized for the detection and retention-time marking of selenium-containing peptides in nanoHPLC (75 μm column) after on-line 100-fold preconcentration on a capillary (300 μm id) precolumn. The mobile phase composition, gradient and flow rate were chosen to allow electrospray-MS/MS to be successfully run in parallel in identical separation and preconcentration conditions in order to produce two matching sets of chromatograms: an element-specific one and a molecule-spe...
Direct comparison of capillary electrophoresis and capillary liquid chromatography hyphenated to collision-cell inductively coupled plasma mass spectrometry for the investigation of Cd-, Cu- and Zn-containing metalloproteins
Abstract Capillary liquid chromatography (cLC) and capillary electrophoresis (CE) have been critically compared for the separation of metalloproteins when using collision-cell inductively coupled plasma mass spectrometry (ICP-CC-MS) as detection system. For cLC separation, the selected column was a C 8 (0.3 mm I.D.) and the separation conditions involved a gradient up to 80% methanol in 10 mM ammonium acetate buffer (pH 7.4). The low flow rate used (3 μL min −1 ) permitted the utilization of a h...
A novel nebulizer (nDS-200) working at sample uptake rates of less than 500 nL min-1 was developed for a sheathless interfacing of nanoHPLC (75-μm column i.d.) with ICPMS. It was based on a hollow fused-silica needle of which the tip (i.d. 10 μm, o.d. 20 μm) centered in a 254-μm-i.d. sapphire orifice. The nebulizer, equipped with a 3-cm3 drain-free vaporization chamber, enabled a stable introduction into an ICP of aqueous mobile phases containing up to 95% acetonitrile at eluent flow rates betwe...
Speciation of selenocompounds by capillary HPLC coupled with ICP-MS using multi-mode gel filtration columns
Multi-mode gel filtration HPLC columns of capillary size coupled with ICP-MS were evaluated as to the separation of naturally occurring selenocompounds. The major selenocompounds found in selenized garlic, i.e., Se-methylselenocysteine and γ-glutamyl-Se-methylselenocysteine, were separated on the narrower column (GS320A-M5D, 0.5 mm i.d. × 150 mm). The selenometabolites found in the urine of a selenium-toxicosis rat, i.e., selenosugar (1-β-methylseleno-N-acetyl-D-galactosamine) and trimethylselen...
Measurement of radical-scavenging ability in hepatic metallothionein of rat using in vivo electron spin resonance spectroscopy.
Abstract In this study, the ability of metallothionein (MT) to scavenge free radicals was determined by in vivo electron spin resonance (ESR) spectroscopy using a carbamoyl-PROXYL, nitroxyl radical, as a spin probe. Production of metallothionein was induced in the liver of rats with ZnSO 4 (0.2 mol/kg, ip) and the intensity of the carbamoyl-PROXYL ESR signal was measured at the upper abdominal level which is a position of the liver. After the injection of carbamoyl-PROXYL, the peak of ESR signal...
Application of CE–ICP–MS and CE–ESI–MS in metalloproteomics: challenges, developments, and limitations
Application of capillary electrophoresis (CE) as a high-resolution separation technique in metalloproteomics research is critically reviewed. The focus is on the requirements and challenges involved in coupling CE to sensitive element and molecule-specific detection techniques such as inductively coupled plasma mass spectrometry (ICP–MS) or electrospray ionisation mass spectrometry (ESI–MS). The complementary application of both detection techniques to the structural and functional characterisat...
Metallothionein transfers zinc to mitochondrial aconitase through a direct interaction in mouse hearts
Abstract Previous studies have shown that in a cell-free system, metallothionein (MT) releases zinc when the environment becomes oxidized and the released zinc is transferred to a zinc-binding protein if such a protein is present. However, it is unknown whether and how zinc transfers from MT to other proteins in vivo. The present study was undertaken to test the hypothesis that if zinc transfer from MT to other proteins occurs in vivo, the transfer would proceed through a direct interaction betw...
Advances in analytical methodology for bioinorganic speciation analysis: metallomics, metalloproteomics and heteroatom-tagged proteomics and metabolomics
The recent developments in analytical techniques capable of providing information on the identity and quantity of heteroatom-containing biomolecules are critically discussed. Particular attention is paid to the emerging areas of bioinorganic analysis including: (i) a comprehensive analysis of the entirety of metal and metalloid species within a cell or tissue type (metallomics), (ii) the study of the part of the metallome involving the protein ligands (metalloproteomics), and (iii) the use of a ...
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: Selenium (Se) belongs to the same group as sulfur in the periodic table but possesses certain chemical properties characteristic of a metal. It is an essential element in animals but becomes severely toxic when the amount ingested exceeds the required level. On the other hand, Se is not essential in plants although some plants are Se hyperaccumulators. Se changes into several chemical forms when metabolized. Thus, the identification of selenometabolites would enable us to formulate a metabolic...
As a result of the greater consciousness within the analytical community on the impact of chemicals on human health and environment, green issues are increasingly taken into account when choosing an established analytical method or developing a new one. Apart from typical analytical characteristics (e.g., sensitivity, limit of detection, repeatability, etc.), other features such as the amount of sample/reagents, operation time, use of energy-effective apparatus, waste production, etc. should be ...