LC/MS/MS Analysis of Immunosuppressants from Whole Blood using Aeris™ Core-Shell HPLC/UHPLC Columns from Phenomenex
28 Oct 2014Cyclosporine A, tacrolimus, sirolimus, and everolimus are four of the most commonly administered immunosuppressant drugs and play a central role in the success of tissue and organ transplants. These drugs are most typically analyzed from whole blood using LC/MS/MS. However, because of the analytical challenges posed when working with whole blood, many of the published methods rely upon complex and/or expensive extraction steps prior to use of the actual analytical column. This application note presents a rapid and effective method for the analysis of these four immunosuppressants from whole blood that uses a simple protein precipitation step followed by direct injection onto a wide-pore core-shell HPLC column (Aeris WIDEPORE 3.6 μm XB-C18). The method displays excellent accuracy and is sensitive down to the low μg/L (ng/mL) range.
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Aeris Core-Shell LC Columns for Proteins and Peptides
Phenomenex IncCore-Shell Particles Precision Engineered for Protein and Peptide Separations. Core-shell particle technology provides striking increases in peak capacity and resolution at lower backpressure, giving chromatographers the ability to achieve ultra-high performance on ANY system, HPLC or UHPLC. A uniform porous silica layer is grown around a solid, spherical silica core, providing effective retention and selectivity with improved resolution, speed, and recovery. Next, optimizing the pore size and shell thickness for intact proteins or smaller peptide fragments provides well-defined depth penetration of biomolecules leading to maximum separation power. Column Specifications: Phase: PEPTIDE XB-C18, WIDEPORE C4, WIDEPORE XB-C18, WIDEPORE XB-C8 Particle Size: 1.7, 2.6, 3.6, 5.0 µm Pore Size: 200, 100 Å Length: 50, 100, 150, 250 mm ID: 2.1, 3.0, 4.6, 10.0, 21.2 PEPTIDE XB-C18 Synthetic peptide impurity analysis Peptide mapping Identifying protein modifications - Glycosylation - Substitution - Truncation Analyzing post-translational modifications - Deamidation - Oxidation - Deletions WIDEPORE XB-C18 Proteins Hydrophilic proteins PEGylated proteins High temperature separations Alternative selectivity for peptide mapping WIDEPORE XB-C8 Large proteins Moderately hydrophobic proteins Monoclonal antibodies Glycosylated proteins High temperature separations WIDEPORE C4 Very large proteins Very hydrophobic proteins Membrane proteins Least retentive