Stephen A Brose1, Mikhail Y Golovko. 1. Department of Pharmacology, Physiology, and Therapeutics, University of North Dakota, Grand Forks, ND 58202-9037, USA.
Abstract
RATIONALE: Fatty acids (FA) labeled with (18) O at the carboxyl group, including oxidized species (FA(18) O), are a useful, low-cost, and easy to prepare tool for quantitative and qualitative mass spectrometry (MS) analysis in biological systems. In addition, they are used to trace the fate of FAs in metabolic pathways including FA re-esterification and lipid remodeling pathways. Although a rapid (18) O exchange on FA(18) O in biological systems has been reported, the mechanism contributing to (18) O exchange has not been fully evaluated. This gap in knowledge limits the use of FA(18) O as a standard for MS and complicates data interpretation for FA metabolism in biological systems. METHODS: In the present study we have addressed a number of possible mechanisms for a rapid (18) O exchange on prostaglandin E(2) (PGE(2) ) using rat plasma as a model. High-performance liquid chromatography coupled with electrospray ionization triple quadrupole MS in the multiple reaction monitoring mode was used for quantification. RESULTS: The major mechanism for a rapid (18) O exchange on PGE(2) (18) O in rat plasma is PGE(2) processing with esterases, while FA re-esterification and non-enzymatic mechanisms do not significantly contribute to this phenomenon. In addition, we report a highly effective inhibition of (18) O exchange with diethylumbelliferyl phosphate that can be used to stabilize FA(18) O in biological samples. CONCLUSIONS: These data indicate the necessity to consider esterase activity when FA(18) O are used to study FA metabolism, and the importance of esterase activity inhibition when FA(18) O are used as internal standards for MS analysis in biological systems. In addition, the results provide a rational for the development of new approaches to study esterase activities and affinity towards modified FA.
RATIONALE: Fatty acids (FA) labeled with (18) O at the carboxyl group, including oxidized species (FA(18) O), are a useful, low-cost, and easy to prepare tool for quantitative and qualitative mass spectrometry (MS) analysis in biological systems. In addition, they are used to trace the fate of FAs in metabolic pathways including FA re-esterification and lipid remodeling pathways. Although a rapid (18) O exchange on FA(18) O in biological systems has been reported, the mechanism contributing to (18) O exchange has not been fully evaluated. This gap in knowledge limits the use of FA(18) O as a standard for MS and complicates data interpretation for FA metabolism in biological systems. METHODS: In the present study we have addressed a number of possible mechanisms for a rapid (18) O exchange on prostaglandin E(2) (PGE(2) ) using rat plasma as a model. High-performance liquid chromatography coupled with electrospray ionization triple quadrupole MS in the multiple reaction monitoring mode was used for quantification. RESULTS: The major mechanism for a rapid (18) O exchange on PGE(2) (18) O in rat plasma is PGE(2) processing with esterases, while FA re-esterification and non-enzymatic mechanisms do not significantly contribute to this phenomenon. In addition, we report a highly effective inhibition of (18) O exchange with diethylumbelliferyl phosphate that can be used to stabilize FA(18) O in biological samples. CONCLUSIONS: These data indicate the necessity to consider esterase activity when FA(18) O are used to study FA metabolism, and the importance of esterase activity inhibition when FA(18) O are used as internal standards for MS analysis in biological systems. In addition, the results provide a rational for the development of new approaches to study esterase activities and affinity towards modified FA.
Authors: H Li; J A Lawson; M Reilly; M Adiyaman; S W Hwang; J Rokach; G A FitzGerald Journal: Proc Natl Acad Sci U S A Date: 1999-11-09 Impact factor: 11.205
Authors: Harpreet Kaur; Kumi Nagamoto-Combs; Svetlana Golovko; Mikhail Y Golovko; Marilyn G Klug; Colin Kelly Combs Journal: Neurobiol Aging Date: 2020-04-18 Impact factor: 4.673