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Annals of Clinical & Laboratory Science 32:181-187 (2002)
© 2002 Association of Clinical Scientists

A New Method for Measuring Oxidative Stress in Claudicants during Strenuous Exercise using Free Radical Derivatives of Antipyrine as Indicators: a Pilot Study.

Stefan A. J. Coolen1, Marc H. W. A. Wijnen2, Jetse C. Reijenga1, Huib L. Vader1,3, Rudi M. H. Roumen1 and Fred A. Huf1
1 University of Technology Eindhoven, Laboratory of Instrumental Analysis, Eindhoven,
2 Department of Paediatric Surgery, University Hospital, Nijmegen, and
3 Department of Surgery, Saint Joseph Hospital, Veldhoven, The Netherlands

Address correspondence to Marc H. W. A. Wijnen, M.D., Department of Paediatric Surgery, University Hospital Nijmegen, P.O. Box 9101, 6500 HB, Nijmegen, The Netherlands; tel 31 49 754 2361; fax 31 24 361 3547; e-mail: mhwijnen{at}worldonline.nl.

Patients with intermittent claudication disease suffer from temporary lack of oxygen in the legs, caused by narrowing of arteries, resulting in ischemia and followed by reperfusion. The degree of oxidative stress present in 16 patients during strenuous exercise was determined using several indicators. Two derivatives of an exogenous marker, antipyrine (AP), (ie, p-hydroxyantipyrine, p-APOH, and o-hydroxyantipyrine, o-APOH), were assayed in plasma using HPLC-tandem-MS. Plasma malondialdehyde (assayed as thiobarb-ituric acid reactive species, TBARS) was also determined. The branchial/ankle blood pressure index (b-a index) was used to assess the severity of intermittent claudication disease, and plasma lactate concentration was also measured as an indicator of the ischemic situation. Plasma TBARS level did not change significantly after exercise. During the ischemic situation as well as during reperfusion, both free radical derivatives of antipyrine increased significantly in plasma (p <0.01). Because p-APOH is also formed enzymatically in humans, the plasma ratio of o-APOH to AP appeared to be the most specific marker for oxidative stress in patients with intermittent claudication.

Keywords: antipyrine, intermittent claudication, malondialdehyde, lactate, HPLC-MS/MS






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