Matrix effects: the Achilles heel of quantitative high-performance liquid chromatography–electrospray–tandem mass spectrometry

High-performance liquid chromatography coupled by an electrospray ion source to a tandem mass spectrometer (HPLC–ESI–MS/MS) is the current analytical method of choice for quantitation of analytes in biological matrices. With HPLC–ESI–MS/MS having the characteristics of high selectivity, sensitivity,...

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Bibliographic Details
Published in:Clinical biochemistry 2005-04, Vol.38 (4), p.328-334
Main Author: Taylor, Paul J.
Format: Article
Language:English
Subjects:
Chromatography, High Pressure Liquid - standards
Electrospray
Ions
Mass Spectrometry - standards
Matrix effects
Physical Phenomena
Physics
Quantitation
Reproducibility of Results
Sensitivity and Specificity
Spectrometry, Mass, Electrospray Ionization - standards
Tandem mass spectrometry
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Summary:High-performance liquid chromatography coupled by an electrospray ion source to a tandem mass spectrometer (HPLC–ESI–MS/MS) is the current analytical method of choice for quantitation of analytes in biological matrices. With HPLC–ESI–MS/MS having the characteristics of high selectivity, sensitivity, and throughput, this technology is being increasingly used in the clinical laboratory. An important issue to be addressed in method development, validation, and routine use of HPLC–ESI–MS/MS is matrix effects. Matrix effects are the alteration of ionization efficiency by the presence of coeluting substances. These effects are unseen in the chromatogram but have deleterious impact on methods accuracy and sensitivity. The two common ways to assess matrix effects are either by the postextraction addition method or the postcolumn infusion method. To remove or minimize matrix effects, modification to the sample extraction methodology and improved chromatographic separation must be performed. These two parameters are linked together and form the basis of developing a successful and robust quantitative HPLC–ESI–MS/MS method. Due to the heterogenous nature of the population being studied, the variability of a method must be assessed in samples taken from a variety of subjects. In this paper, the major aspects of matrix effects are discussed with an approach to address matrix effects during method validation proposed.
ISSN:0009-9120
1873-2933
DOI:10.1016/j.clinbiochem.2004.11.007