Studies on Chromatographic Fractioning by Cations Exchangers of a Bovine Hemoglobin Hydrolysate

2018 ◽  
Vol 69 (10) ◽  
pp. 2794-2798
Author(s):  
Alina Diana Panainte ◽  
Ionela Daniela Morariu ◽  
Nela Bibire ◽  
Madalina Vieriu ◽  
Gladiola Tantaru ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Retention Time ◽  
Chromatographic Separation ◽  
Bovine Hemoglobin ◽  
Reverse Phase ◽  
Hplc System ◽  
Fast Flow ◽  
Hydrolysis Of ◽  
Phase Column

A peptidic hydrolysate has been obtained through hydrolysis of bovine hemoglobin using pepsin. The fractioning of the hydrolysate was performed on a column packed with CM-Sepharose Fast Flow. The hydrolysate and each fraction was filtered and then injected into a HPLC system equipped with a Vydak C4 reverse phase column (0.46 x 25 cm), suitable for the chromatographic separation of large peptides with 20 to 30 amino acids. The detection was done using mass spectrometry, and the retention time, size and distribution of the peptides were determined.

scholarly journals High-Throughput Analysis of Amino Acids for Protein Quantification in Plant and Animal-Derived Samples Using High Resolution Mass Spectrometry

Molecules ◽  
2021 ◽  
Vol 26 (24) ◽  
pp. 7578
Author(s):  
Priyanka Reddy ◽  
Aaron Elkins ◽  
Joe Panozzo ◽  
Simone J. Rochfort
Keyword(s):  
Mass Spectrometry ◽  
Amino Acids ◽  
Retention Time ◽  
High Resolution Mass Spectrometry ◽  
Amino Acid Content ◽  
Protein Quantification ◽  
Borate Buffer Solution ◽  
Buffer Solution ◽  
High Throughput Analysis ◽  
Protein Levels

Current methods for measuring the abundance of proteogenic amino acids in plants require derivatisation, extended run times, very sensitive pH adjustments of the protein hydrolysates, and the use of buffers in the chromatographic phases. Here, we describe a fast liquid chromatography–mass spectrometry (LC–MS) method for the determination of amino acids that requires only three steps: hydrolysis, neutralisation, and sample dilution with a borate buffer solution for pH and retention time stability. The method shows excellent repeatability (repeated consecutive injections) and reproducibility (repeated hydrolysis) in the amino acid content, peak area, and retention time for all the standard amino acids. The chromatographic run time is 20 min with a reproducibility and repeatability of <1% for the retention time and <11% for the peak area of the BSA and quality control (QC) lentil samples. The reproducibility of the total protein levels in the hydrolysis batches 1–4 was <12% for the BSA and the lentil samples. The level of detection on column was below 0.1 µM for most amino acids (mean 0.017 µM).

scholarly journals Non-derivatized glycan analysis by reverse phase liquid chromatography and ion mobility-mass spectrometry

The Analyst ◽  
2015 ◽  
Vol 140 (10) ◽  
pp. 3335-3338 ◽  
Author(s):  
Nichole M. Lareau ◽  
Jody C. May ◽  
John A. McLean
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Ion Mobility ◽  
Ion Mobility Mass Spectrometry ◽  
Reverse Phase ◽  
Simple Method ◽  
Glycan Analysis ◽  
Reverse Phase Liquid Chromatography ◽  
Phase Liquid ◽  
Phase Column

A simple method for the analysis of non-derivatized glycans using a reverse phase column on a liquid chromatography-ion mobility-mass spectrometry (LC-IM-MS) instrument.

scholarly journals Hydrolysis of Casein-Derived Peptides αS1-Casein(f1-9) and β-Casein(f193-209) by Lactobacillus helveticus Peptidase Deletion Mutants Indicates the Presence of a Previously Undetected Endopeptidase

2003 ◽  
Vol 69 (2) ◽  
pp. 1283-1286 ◽  
Author(s):  
Jeffrey E. Christensen ◽  
Jeffery R. Broadbent ◽  
James L. Steele
Keyword(s):  
Mass Spectrometry ◽  
High Performance Liquid Chromatography ◽  
High Performance ◽  
Lactobacillus Helveticus ◽  
Deletion Mutants ◽  
Reverse Phase ◽  
Peptide Bonds ◽  
Cell Extracts ◽  
Hydrolysis Products ◽  
Hydrolysis Of

ABSTRACT Peptides derived from hydrolysis of αS1-casein(f1-9) [αS1-CN(f1-9)] and β-CN(f193-209) with cell extracts of Lactobacillus helveticus CNRZ32 and single-peptidase mutants (ΔpepC, ΔpepE, ΔpepN, ΔpepO, and ΔpepX) were isolated by using reverse-phase high-performance liquid chromatography and were characterized by mass spectrometry. The peptides identified suggest that there was activity of an endopeptidase, distinct from previously identified endopeptidases (PepE and PepO), with specificity for peptide bonds C terminal to Pro residues. Identification of hydrolysis products derived from a carboxyl-blocked form of β-CN(f193-209) confirmed that the peptides were derived from the activity of an endopeptidase.

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