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An improved GC/MS-based procedure for the quantitation of the isoprostane 15-F2t-IsoP in rat plasma

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Abstract

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.

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Authors and Affiliations

  1. Laboratory of Structural Biology, National Institute of Environmental Health Sciences, 27709, Research Triangle Park, NC

    Carol E. Parker

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  1. Carol E. Parker
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  2. LeRae B. Graham
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  3. My-Nga Nguyen
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  4. Beth C. Gladen
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  5. Maria B. Kadiiska
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  6. J. Carl Barrett
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  7. Kenneth B. Tomer
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Parker, C.E., Graham, L.B., Nguyen, MN. et al. An improved GC/MS-based procedure for the quantitation of the isoprostane 15-F2t-IsoP in rat plasma. Mol Biotechnol 18, 105–118 (2001). https://doi.org/10.1385/MB:18:2:105

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