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Overcoming the Matrix Effect in LC/MS of Serum or Plasma Samples Using Two Distinct Sample Prep Approaches

Sigma-Aldrich Supelco
19 Apr 2016

This article highlights the impact that sample matrix effects can have on LC/MS response and discusses two novel approaches to reduce it. Advancements in electrospray ionization mass spectrometry (ESI-MS) have changed the way identification and quantification of small molecules in biological fluids is conducted. Systems are capable of routinely detecting sub-femtogram levels of analytes. This report demonstrates that targeted phospholipid depletion using HybridSPE-Phospholipid and analyte enrichment using biocompatible SPME, both provide effective means to reduce matrix interference that can rob the method of sensitivity, reproducibility, precision, and accuracy.

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HybridSPE®-Precipitation, 96-well Plate, pk of 1

Sigma-Aldrich Supelco

Patent pending HybridSPE – Precipitation (HybridSPE-PPT) technology is a simple and generic sample prep platform designed for the gross level removal of endogenous protein and phospholipid interferences from biological plasma and serum prior to LC-MS or LC-MS/MS analysis. Biological plasma or serum is first subjected to protein precipitation via the addition and mixing of acidified acetonitrile. Precipitated proteins are then removed by centrifugation and the resulting supernatant is loaded on the HybridSPE-PPT 96-well plate or cartridge which acts as a chemical filter that specifically targets the removal of endogenous sample phospholipids. The phospholipid retention mechanism is based on a highly selective Lewis acid-base interaction between the proprietary zirconia ions functionally bonded to the HybridSPE-PPT stationary phase and the phosphate moiety consistent with all phospholipids. The resulting eluent is ready for immediate LC-MS or LC-MS-MS analysis. An alternative “In-well Precipitation” method is available for the HybridSPE-PPT 96-well version in which biological plasma/serum is first added to the 96-well plate followed by acidified acetonitrile (precipitation agent). After a brief mixing/vortexing step, vacuum is applied to the 96-well plate. Because the 96-well version contains a series of low porosity hydrophobic filters/frits, the packed-bed filter/frit assembly acts as a depth filter facilitating the concurrent removal of both phospholipids and precipitated proteins during the extraction process.

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Sample PreparationSample preparation can improve the quality and speed of separation techniques. Products to assist sample preparation include filtration equipment, evaporators, membranes and sieves.LC-MSLC-MS (liquid chromatography-mass spectrometry) systems and equipment are used for separation and quantitative analysis of complex mixtures, combining liquid chromatography and mass spectrometry. Quantify proteins, contaminants, pesticides or screen for drug metabolites with a high level of sensitivity. LC-MS systems and equipment include reverse phase, normal phase and specialized columns integrated with various MS detectors such as time-of-flight (TOF), quadrupole, orbitrap or ion trap mass analyzers. LC-MS/MS instruments equipped with a qTOF or triple quadrupole analyzer give greater sensitivity and resolving power to your analysis. Find the best LC-MS equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Mass SpectrometryMass spectrometry (MS) is a powerful analytical technique used to identify and quantify molecules based on the mass-to-charge ratio of gas-phase ions. It provides detailed information about the structure, composition, and properties of compounds and is widely used across fields such as environmental monitoring, materials science, drug discovery and development, food and beverage testing, and wider chemical research. Key MS techniques include tandem mass spectrometry (MS/MS), liquid chromatography–mass spectrometry (LS-MS) and inductively coupled plasma (ICP-MS). Choosing from these wide range of techniques and technologies can be a daunting task, so keep up to date with scientific applications, performance expectations, and customer reviews here all in one place. Visit our product directory to receive quotes direct from the manufacturer. Target-EnrichmentPlasmaHuman SerumHuman serum is found in blood plasma. The protein is used in a variety of applications including tissue engineering, transplantation and cell therapy.Electrospray IonizationLC-MSLiquid Chromatography-Mass Spectrometry (LCMS) is a powerful analytical technique that combines the separation power of liquid chromatography with the detection capabilities of mass spectrometry. It is widely used for qualitative and quantitative analysis of complex mixtures in pharmaceuticals, proteomics, and environmental studies. Browse our peer-reviewed product directory to find the best LCMS systems, compare products, check reviews, and get pricing directly from manufacturers.

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