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Challenges in accurate quantitation of lysophosphatidic acids in human biofluids

Authors :
Anne-Françoise Aubry
Petia Shipkova
Anne Minnich
John A. Easter
Adrienne A. Tymiak
Joelle M. Onorato
Source :
Journal of Lipid Research, Vol 55, Iss 8, Pp 1784-1796 (2014)
Publication Year :
2014
Publisher :
The American Society for Biochemistry and Molecular Biology, 2014.

Abstract

Lysophosphatidic acids (LPAs) are biologically active signaling molecules involved in the regulation of many cellular processes and have been implicated as potential mediators of fibroblast recruitment to the pulmonary airspace, pointing to possible involvement of LPA in the pathology of pulmonary fibrosis. LPAs have been measured in various biological matrices and many challenges involved with their analyses have been documented. However, little published information is available describing LPA levels in human bronchoalveolar lavage fluid (BALF). We therefore conducted detailed investigations into the effects of extensive sample handling and sample preparation conditions on LPA levels in human BALF. Further, targeted lipid profiling of human BALF and plasma identified the most abundant lysophospholipids likely to interfere with LPA measurements. We present the findings from these investigations, highlighting the importance of well-controlled sample handling for the accurate quantitation of LPA. Further, we show that chromatographic separation of individual LPA species from their corresponding lysophospholipid species is critical to avoid reporting artificially elevated levels. The optimized sample preparation and LC/MS/MS method was qualified using a stable isotope-labeled LPA as a surrogate calibrant and used to determine LPA levels in human BALF and plasma from a Phase 0 clinical study comparing idiopathic pulmonary fibrosis patients to healthy controls.

Details

Language :
English
Database :
OpenAIRE
Journal :
Journal of Lipid Research, Vol 55, Iss 8, Pp 1784-1796 (2014)
Accession number :
edsair.doi.dedup.....c59e12919e197f8d56b2c1254af4c77b