Temperature and time-dependent effects of delayed blood processing on oxylipin concentrations in human plasma.
Author(s): Ramsden CE, Yuan ZX, Horowitz MS, Zamora D, Majchrzak-Hong SF, Muhlhausler BS, Taha AY, Makrides M, Gibson RA
Publication: Prostaglandins Leukot Essent Fatty Acids, 2019, Vol. 150, Page 31-37
PubMed ID: 31568925 PubMed Review Paper? No
Purpose of Paper
This paper investigated the effects of pre-centrifugation storage of blood at room temperature or on wet ice on levels of 16 oxylipins and thromboxane B2 (TXB2) in the plasma of a single volunteer.
Conclusion of Paper
Levels of 12-lipoxygenase (HETE), TXB2, and 14-hydroxy-docohexaenoic acid (HDHA) increased with centrifugation delays at room temperature but were unaffected by centrifugation delays on wet ice resulting in statistically significant differences after 120 min. Levels of 5-HETE, 15-HETE, 4-HDHA, 10-HDHA, 17-HDHA, 19,20- epoxy-docosapentaenoic acid (EpDPA), and 9-hydroxyoctadecadienoic acid (HODE) were not affected by centrifugation delays at room temperature or on ice. Levels of prostaglandin E2 (PGE2) and 8- isoprostane (8-isoPGF2α) were below the limit of quantification (LOQ) in all specimens.
Studies
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Study Purpose
This study investigated the effects of pre-centrifugation storage of blood at room temperature or on ice on levels of 16 oxylipins and TBX2 in plasma of a single volunteer. EDTA blood was obtained using an 18-gauge butterfly needle from a single fasting volunteer and stored on ice or at room temperature. After storage, plasma was obtained by centrifugation at 1880 x g for 10 min. Lipid extracts were purified using Strata X cartridges and eluted in methanol. After evaporation and reconstitution, oxylipins were quantified by ultra-performance liquid chromatography (UPLC) tandem electrospray negative ionization mass spectrometry. The experiment was run in duplicate on three separate days such that there were six specimens for each timepoint. The LOQ was defined as a signal-to-noise ratio >5. Levels below the LOQ were imputed as being 50% the LOQ.
Summary of Findings:
Levels of 12-HETE, TXB2, and 14-HDHA increased with centrifugation delays at room temperature but were unaffected by centrifugation delays on wet ice resulting in significant differences between specimens stored at the two temperatures after 120 min (P=0.005, P=0.009, and P=0.001, respectively). Levels of 5-HETE,15-HETE, 4-HDHA, 10-HDHA, 19,20-EpDPA, and 9-HODE were not affected by centrifugation delays at room temperature or on ice. Levels of 17-HDHA varied during the course of the study but variance did not seem to depend on storage. Levels of PGE2 and 8-isoPGF2α were below the LOQ in all specimens.
Biospecimens
Preservative Types
- None (Fresh)
Diagnoses:
- Not specified
Platform:
Analyte Technology Platform Small molecule LC-MS or LC-MS/MS Lipid LC-MS or LC-MS/MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(s) Biospecimen Aliquots and Components Centrifugation Centrifugation delays investigated
Storage Storage temperature Room temperature
Wet ice
Storage Storage duration 0 min
10 min
20 min
30 min
60 min
120 min