An improved GC/MS-based procedure for the quantitation of the isoprostane 15-F2t-IsoP in rat plasma.
This article describes a procedure for the quantitation of the isoprostane 15-F2t-IsoP (9a,11a,15S-trihydroxy-(8b)-prosta-5Z,13E-dien-1-oic acid [CAS#27415-26-5] formerly known as 8-epi-PGF2a or 8-iso-PGF2a, and also as iPF2a-III). We have combined features from several earlier methods for 15-F2t-IsoP and prostaglandins, and identified and modified those steps that may lead to poor recoveries. The resulting protocol is precise and reliable, and was validated by a blind time-course study of plasma levels in rats treated with 120 and 1200 mg CCl4/kg body weight. Plasma levels of 15-F2t-IsoP, as measured according to the procedure described above, are good indicators of acute oxidative stress as induced by CCl4. The precision of the measurements allows detection of elevated plasma 15-F2t-IsoP levels as long as 16 h after an acute exposure of 120 mg CCl4/kg body weight, and 2 h after an exposure of 1 mg CCl4/kg body weight. The results of this low-dose, pilot study suggest that this method has sufficient analytical precision to allow the detection of the small changes in plasma isoprostane levels, which result from chronic and/or lower-level exposures to agents causing oxidative stress.
Parker
C E
CE
Laboratory of Structural Biology, National Institute of Environmental Health Sciences, Research Triangle Park, NC 27709, USA.
Graham
L B
LB
Nguyen
M N
MN
Gladen
B C
BC
Kadiiska
M B
MB
Barrett
J C
JC
Tomer
K B
KB
D000818:Animals; D001835:Body Weight; D002251:Carbon Tetrachloride / Q000494:pharmacology; D015237:Dinoprost / Q000031:analogs & derivatives* / Q000097:blood* / Q000737:chemistry* / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006868:Hydrolysis; D008297:Male; D018384:Oxidative Stress / Q000187:drug effects; D010865:Pilot Projects; D000074262:Rapeseed Oil; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D005229:Fatty Acids, Monounsaturated; D000074262:Rapeseed Oil; C075750:8-epi-prostaglandin F2alpha; D015237:Dinoprost; D002251:Carbon Tetrachloride