Increase of tissue lipid hydroperoxides as determination of oxidative stress

Increased levels of lipid hydroperoxides (LOOH) are frequently associated w ith the oxidative mechanisms involved in physiological states as ageing and with serious pathological conditions. In the present work the physiologica l and the CCl4-induced lipid hydroperoxides levels in mice liver and kidney were determined. The analysis of LOOH tissue levels was performed through the oxidation of 1-napthyldiphenylphosphine (NDPP) into its oxide (ONDPP) a nd further quantification by high pressure liquid chromatography at 292 nm UV detection. The physiological level of lipid hydroperoxides levels was higher in the ki dney (245 +/- 8 nmol LOOH/ g of tissue) than in liver ( 164 +/- 5 nmol of L OOH/ g tissue). After a single administration of CCl4 (0.25 ml/ g) tissue L OOH reached a maximum level after 15 min (416 +/- 21 nmol/g kidney and 303 +/- 6 nmol/ g liver) and then slowly decreased. LOOH levels in liver afford ed an early indicator (15 min) of oxidative damage. LOOH levels in kidney r emained significatively increased up to 60 min post administration. The described HPLC assay is a useful, simple and sensitive method to detect cellular oxidative stress and damage.