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Quantification Purposes (quantification + purpose)
Selected AbstractsChemistry meets proteomics: The use of chemical tagging reactions for MS-based proteomicsPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 20 2006Alexander Leitner Dr. Abstract As proteomics matures from a purely descriptive to a function-oriented discipline of the life sciences, there is strong demand for novel methodologies that increase the depth of information that can be obtained from proteomic studies. MS has long played a central role for protein identification and characterization, often in combination with dedicated chemical modification reactions. Today, chemistry is helping to advance the field of proteomics in numerous ways. In this review, we focus on those methodologies that have a significant impact for the large-scale study of proteins and peptides. This includes approaches that allow the introduction of affinity tags for the enrichment of subclasses of peptides or proteins and strategies for in,vitro stable isotope labeling for quantification purposes, among others. Particular attention is given to the study of PTMs where recent advancements have been promising, but many interesting targets are not yet being addressed. [source] Quantification of Greenland halibut serum vitellogenin: a trip from the deep sea to the mass spectrometerRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 7 2009Alejandro M. Cohen This paper focuses on the sequential steps involved in developing a technique for quantifying Greenland halibut vitellogenin, a serum protein biomarker, using a comprehensive mass spectrometric approach. In the first phase of this study, in-gel trypsin digestions of serum proteins separated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) were analyzed by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). A characteristic band around a molecular mass of 185,kDa, present in the mature female specimens, but absent in the male samples, was identified as vitellognin according to the peptide mass fingerprint obtained by MALDI-MS. Subsequently, MALDI and electrospray ionization tandem mass spectrometry (ESI-MS/MS) analyses were performed on the digest of the vitellogenin band for de novo sequencing. From these studies, a characteristic 'signature' peptide (sequence: FFGQEIAFANIDK) was selected from a list of candidate peptides as a surrogate analytical standard used for quantification purposes. Sample preparation for vitellogenin quantification consisted of a simple one-step overnight trypsin digestion. Samples were spiked with an isotopologue signature peptide standard and analyzed by high-performance liquid chromatography (HPLC) coupled in-line to an electrospray quadrupole-hexapole-quadrupole tandem mass spectrometer, operated in selective reaction monitoring mode. Transitions [(m/z 750.0,,,1020.4 and 750.0,,,1205.4) and (754.8,,,1028.6 and 754.8,,,1213.2)] were monitored for the signature peptide and the internal standard, respectively. Samples obtained from the field showed that vitellogenin levels were in accordance with fish maturity determined by macroscopic examination of the gonad, proving this technique suitable for measuring vitellogenin as a serum protein biomarker for reproductive maturity in female fish. Copyright © 2009 John Wiley & Sons, Ltd. [source] Imaging of uranium on rat brain sections using laser ablation inductively coupled plasma mass spectrometry: a new tool for the study of critical substructures affined to heavy metals in tissuesRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 18 2008J. Sabine Becker The specific toxicity of trace metals and compounds largely depends on their bioavailability in different organs or compartments of the organism considered. Imaging mass spectrometry (IMS) using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) with a spatial resolution in the 100,µm range was developed and employed to study heavy metal distribution in brain tissues for toxicological screening. Rat brain post-mortem tissues were stained in an aqueous solution of either uranium or neodymium (metal concentration 100,µg,g,1) for 3,h. The incubation of heavy metal in thin slices of brain tissue is followed by an imaging mass spectrometric LA-ICP-MS technique. Stained rat brain tissue (thickness 30,µm) were scanned with a focused laser beam (wavelength 266,nm, diameter of laser crater 100,µm and laser power density 3,×,109,W,cm,2). The ion intensities of 235U+, 238U+, 145Nd+ and 146Nd+ were measured by LA-ICP-MS within the ablated area. For quantification purposes, matrix-matched laboratory standards were prepared by dosing each analyte to the pieces of homogenized brain tissue. Imaging LA-ICP-MS allows structures of interest to be identified and the relevant dose range to be estimated. Copyright © 2008 John Wiley & Sons, Ltd. [source] Multiresidue determination of antibiotics in aquaculture fish samples by HPLC,MS/MSAQUACULTURE RESEARCH, Issue 9 2010Consuelo Cháfer-Pericás Abstract An analytical method based on HPLC with MS/MS detection was developed and optimized in order to determine the most useful antibiotics (sulphonamides and tetracyclines) used in aquaculture. A simple extraction procedure, without any clean-up step, was evaluated in order to obtain maximum analyte recovery from fish samples (Sparus aurata). A mixture of methanol:water 70:30 (v/v)+1 mL EDTA 0.1 M was selected as optimum extractant solution. Because no matrix effects were observed, a standard calibration curve prepared in mobile phase was used for quantification purposes. Antibiotic-free fish samples were spiked at different concentration levels and analysed by the optimized HPLC method. The average recoveries (n=6) obtained were satisfactory, ranging from 88% to 110% at 100 ,g kg,1. The proposed methodology provided limits of detection for the tested antibiotics in the 1.2,16 ,g kg,1 range, lower than 100 ,g kg,1, the maximum residue level established by the European Union. Finally, commercial fish samples from different origins were analysed in order to confirm the usefulness of the developed methodology. [source] | ||||||||||