bioresources, production of industrially important biomolecules, reduction of environmental pollution but will also reveal promising therapeutic utilization of this by-product. Especially, structural elucidation of oligosaccharides from apple pomace enables the understanding and prediction of their biological functions. High resolution mass spectrometry and tandem mass spectrometry provides a powerful and highly sensitive analytical tool to improve the oligosaccharides structural knowledge. Here, we report the quantification of main monosaccharides, and isolation and structural characterization of a pool of oligosaccharides in apple pomace. Free monosaccharides and oligosaccharides were extracted with water under room temperature or boiling temperature, and the profiles under these two different extraction conditions were compared. Monosaccharides were quantified by high performance anion-exchange chromatography with pulse amperometric detection (HPAEC-PAD) and enzymatic kit. Oligosaccharides were characterized with liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF), and further structural information was obtained through tandem MS/MS.…
Introduction: One of the most common mass spectrometry–based proteomic methodologies used to day is the bottom up protein identification. First, the protein has to be digested into peptides using a protease. Then, the peptide will be submitted to the URMC Proteomics Resource and Laboratory for analysis using LC-MS/MS tandem mass spectrometry. In this approach, the peptides will be separated and introduced into a mass spectrometer using reversed phase liquid chromatography (RC-LC). This…
for 30 min at room temperature. Then, trypsin/chymotrypsin- digested samples of KPNB1 obtained from the UV-crosslinking experiments with various concentrations of compound 1 were analyzed by nanoflow liquid chromatography and tandem mass spectrometry. The tandem mass spectra of the trypsin/chymotrypsin-digested KPNB1-compound 1 adducts showed 10 peptides coupled to compound 1 (Table S1, the Supplementary Information). Among these adducts, we found that the amounts of three of the products,…
the analysis making a suitable sample preparation and extraction process vital. In an attempt to resolve these issues, liquid chromatography–tandem mass spectrometry method is reported for the determination of methamphetamine and amphetamine in human hair with anion exchange solid phase extraction. 2. LIQUID CHROMATOGRAPHY-TANDEM MASS SPECTROMETRY (LC-MS) Liquid chromatography is the most widely used technique for the analysis of chemical mixtures. In order to separate, identify and quantify…
phase, which is essential for separating charged compounds such as acidic oligosaccharides58. Structural characterization using mass spectrometry High resolution mass spectrometry (MS) and tandem mass spectrometry provide a powerful and highly sensitive analytical tool to obtain the structural information of oligosaccharides. Therefore, they are now accepted as an ideal tool to couple with HPLC for the oligosaccharide characterization in various types of samples. The mass spectrometric…
transferring the mixed protein solution to isoelectric tube gel (pH 3-10) and subjecting it to a constant electric current (145mA). After the gel has been allowed to run for about an hour, the isoelectric tube will be rinsed with running buffer and set aside. Next, an SDS gel will be prepared in order to separate the proteins based on mass. This will be done by sandwiching two spacers between two glass plates and placing them within a plastic bag. The plate/bag assembly will then be placed into…
The role of the phenomenon of postmortem redistribution in forensic cases Abstract It has been established in several studies that a postmortem blood sample examination is not always a reliable reflect of blood sample examination during life. There are a large number of variables affecting the blood concentration of foreign substances in a living individual, in the other hand, there are also a number of factors that can change their levels after death. Although the underlying mechanisms involved…
a Beckman DU650 spectrophotometer (Beckman Coulter, Fullerton, USA). Phenolic compounds were isolated using a multiple preparation HPLC system (LC-Forte/R, YMC, Kyoto, Japan) with a reverse phase preparative column (YMC-Actus Triart Prep C18, 20 mm × 250 mm, 10 μm). 1H and 13C NMR data were obtained usingn a Bruker AM 500 (1H NMR at 500 MHz, 13C NMR at 125 MHz) spectrometer (Bruker, Karlsruhe, Germany) using DMSO-d6 with TMS as the internal standard. For the quantification of the isolated…
Aim: Discuss the advantages, disadvantages and limitations of mass spectrometry for the analysis of both small molecules and proteins. Background Mass spectrometers. They are a powerful tool used to evaluate analytes such as single atoms, molecules, whole proteins, or the simple peptide chains that make them up. Using the mass to charge ratio (m/z) of these analytes, we can differentiate one or many entities (such as one particular peptide fragment, or a panel of them) from everything else in…
The liquid chromatography was performed using an Acquity UPLC® system (Waters Corp., Milford, MA, USA) coupled to a mass spectrometer (Xevo TQ-S, Waters Corp.). The bioanalytical method for the simultaneous quantitation of the three PFCs was optimized through the various mobile phase conditions and sample preparation processes. A 3 µL aliquot of each sample was injected into a Kinetex® 2.6 µm C8 100 Å column (2.1 mm × 100 mm, 2.7 µm particle size, Phenomenex, Inc., USA) with a column…