LC-MS/MS Determination of Isoprostanes in Plasma Samples Collected from Mice Exposed to Doxorubicin or Tert-Butyl Hydroperoxide

International Journal of Molecular Sciences

Monika Janicka²  Agata Kot-Wasik²  Jolanta Paradziej-Łukowicz³  Grażyna Sularz-Peszyńska³  Agnieszka Bartoszek¹ 

[1] Department of Food Chemistry, Technology and Biotechnology, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland; E-Mail:;Department of Analytical Chemistry, Chemical Faculty, Gdańsk University of Technology, Narutowicza 11/12, 80-233 Gdańsk, Poland; E-Mails:;Tri-City Central Animal Laboratory Research and Service Centre of the Medical University of Gdansk, Medical University of Gdansk, 80-210 Gdańsk, Poland; E-Mails:

关键词: isoprostanes;    doxorubicin;    plasma biomarkers of oxidative stress;    LC-MS/MS;   

DOI  :  10.3390/ijms14036157

来源: mdpi

【 摘 要 】

Isoprostanes are stable products of arachidonic acid peroxidation and are regarded as the most reliable markers of oxidative stress in vivo. Here we describe the LC-MS/MS procedure enabling simultaneous determination of four regioisomers (8-iso prostaglandin F_2α, 8-iso-15(R)-prostaglandin F_2α, 11β-prostaglandin F_2α, 15(R)-prostaglandin F_2α) in plasma samples collected from mice. The four plasma isoprostanes are determined by LC–ESI-MS/MS with deuterated 8-iso-PGF_2α-d₄ as an internal standard (I.S.). For plasma samples spiked with the isoprostanes at a level of 200 pg/mL each, the method imprecision has been below 7.1% and mean inaccuracy equaled 8.7%. The applicability of the proposed approach has been verified by the assessment of changes in isoprostane levels in plasma samples derived from mice exposed to tert-butyl hydroperoxide (TBHP), a model inducer of oxidative stress, or to antitumor drug doxorubicin (DOX) known for potent stimulation of redox cycling. Compared to the control group of mice, both oxidative stress inducers tested increased the levels of three out of four isoprostanes in exposed animals; 11β-prostaglandin F_2α being the exception. The greatest rise was observed in the case of 15(R)-prostaglandin F_2α, by about 50% and 70% in plasma samples derived from mice exposed to DOX and TBHP, respectively.

【 授权许可】

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© 2013 by the authors; licensee MDPI, Basel, Switzerland.