An Improved Procedure for the Enrichment of Plasma F2-Isoprostanes Prior to Final Determination by GC-MS/NICI
نویسندگان
1 Department of Biochemistry,Urmia University
doi
چکیده
One of the most popular approaches to quantify oxidative injury is to measure lipid peroxidation products and, in particular, F2-isoprostanes (F2-IPs). F2-IPs is a group of prostaglandin F2-like compounds derived from the non-enzymatic oxidation of arachidonic acid. Of these, the 15-F2t-isoprostane (8-iso-PGF2 ) has received considerable attention, because it possesses adverse biological activities. Previous Gas Chromatographic-Mass Spectrometric (GC-MS) methods for measuring plasma F2-IPs from this laboratory involved two chromatography steps on C18 and NH2-cartridges. Problems may, however, arise with chromatography on C18 cartridges, as it can be time-consuming and losses may occur depending upon the pH and eciency of the sample loading. Therefore, it was decided that the C18 chromatography step be replaced with a single lipid partitioning step and the NH2-chromatography be simpli ed. In 70 plasma samples from healthy individuals, total (sum of free and esteri ed) 15-F2t-isoprostane concentrations ranged from 0.5 to 3.13 nM. This assay meets all prede ned method performances in terms of speci city and sensitivity. The improved method is suitable for the analysis of samples from larger clinical trials investigating the role of oxidant injury under conditions associated with oxidative stress. One of the most popular approaches to quantify oxidative injury is to measure lipid peroxidation products and, in particular, F2-isoprostanes (F2-IPs). F2-IPs is a group of prostaglandin F2-like compounds derived from the non-enzymatic oxidation of arachidonic acid. Of these, the 15-F2t-isoprostane (8-iso-PGF2 ) has received considerable attention, because it possesses adverse biological activities. Previous Gas Chromatographic-Mass Spectrometric (GC-MS) methods for measuring plasma F2-IPs from this laboratory involved two chromatography steps on C18 and NH2-cartridges. Problems may, however, arise with chromatography on C18 cartridges, as it can be time-consuming and losses may occur depending upon the pH and eciency of the sample loading. Therefore, it was decided that the C18 chromatography step be replaced with a single lipid partitioning step and the NH2-chromatography be simpli ed. In 70 plasma samples from healthy individuals, total (sum of free and esteri ed) 15-F2t-isoprostane concentrations ranged from 0.5 to 3.13 nM. This assay meets all prede ned method performances in terms of speci city and sensitivity. The improved method is suitable for the analysis of samples from larger clinical trials investigating the role of oxidant injury under conditions associated with oxidative stress.
