What do thiobarbituric acid and hemoglobin-methylene blue tests evaluate in the endogenous lipid peroxidation of rat liver?

Abstract

SummaryTo detect the endogenous lipid peroxides in rat liver, the thiobarbituric acid (TBA) and hemoglobin-methylene blue (HMB) tests were used. The endogenous lipid peroxidation in rat liver was induced by peroxidative drugs, and then these tests were carried out. When CC14, paraquat or phenobarbital was intraperitoneally injected, the HMB values were significantly increased only at the early period after the doses (1 h), whereas two peaks of TBA value were observed at 1 h and around 24 h after the doses. When peroxidation was induced by thiopental or alcohol, the TBA value also gave two peaks. It was suggested that the agreement between detections by both HMB and TBA (the early peak) indicated the peroxide formation, and that the independent increase in TBA value (second peak) the accumulation of aldehydes in the liver. The life span of HMB-reactive substances was short in vivo compared with that of the TBA-reactive substances. It is considered that the TBA test was useful to detect the endogenous lipid peroxidation under the conditions that TBA test could detect aldehydes originating from lipid peroxidation because lipid peroxides were unstable in vivo.

Circulating cytokines and hormones with immunosuppressive but neutrophil-priming potentials rise after endurance exercise in humans

“…Therefore, we used the HMB test to measure LPO, which has been shown to be more speci®c for LPO than the TBARS test (Kanazawa et al 1989). In spite of the marked rise of FFA after the marathon race, we could not obtain positive data, which is entirely consistent with the reported literature using the TBARS test (Dufaux et al 1996;Margaritis et al 1997;értenblad et al 1997).…”

“…In spite of the marked rise of FFA after the marathon race, we could not obtain positive data, which is entirely consistent with the reported literature using the TBARS test (Dufaux et al 1996;Margaritis et al 1997;értenblad et al 1997). This may be related to the fact that LPO have been found to be unstable and their life spans shorter in vivo (Kanazawa et al 1989), that other free radical scavenging mechanisms such as glutathione redox have been shown to work eectively and compensate for the development of lipid peroxidation, and that trained athletes possess a greater capacity of these scavenger and antioxidant systems compared with sedentary individuals (Dufaux et al 1996).…”

“…Lipid peroxidation has been evaluated by measuring malondialdehyde (MDA) concentrations using the thiobarbituric acid reactive substances (TBARS) assay, but this method has often been criticized for its indirect reaction with LPO and lack of speci®city for free MDA formed in vivo (Kanazawa et al 1989;Dufaux et al 1996). Therefore, we used the HMB test to measure LPO, which has been shown to be more speci®c for LPO than the TBARS test (Kanazawa et al 1989).…”

“…The PRL concentration was measured using immunoradiometric assay (SPAC-S prolactin kit, Daiichi Radioisotope Institute, Tokyo, Japan). Lipid peroxides (LPO) concentration were assessed using a haemoglobin-methylene blue (HMB) test kit (Determiner LPO, Kyowa Medics, Tokyo, Japan; Kanazawa et al 1989). The NO concentration was assessed by measuring total nitrite/nitrate concentrations in plasma with a Griess reagent kit (NO 2 /NO 3 Assay kit-C, Dojindo, Kumamoto, Japan) after removal of plasma proteins by ultra®ltration using Ultrafree-MC 10,000 NMWL Filter Unit (Millipore Co., Bedford, Ma, USA).…”

Ascorbic acid prevents ischemia-reperfusion injury in the rat small intestine

“…When hemoglobin reduces peroxides to their corresponding alcohols, N-methylcarbamoyl derivatives of methylene blue (leuco form) are oxidized and colored blue [11].…”