Do we need to use the seminal plasma alone or the whole sperme or sprme cells only without seminal plasma?
dear hamza
Total antioxidant activity was determined spectrophotometrically according to the ability of seminal plasma antioxidants to scavenge the 2,2′-azinobis(3-ethylbenzothiazoline 6-sulphonate) (ABTS) radical cation, ABTS+, inhibiting its absorption at 734 nm (Rice-Evans and Miller, 1994; Miller and Rice-Evans, 1996).
Semen (0.5 ml) was centrifuged at 3000 g for 10 min. An 8.4 μl aliquot of the supernatant was added to a solution containing 2.5 μmol/l metmyoglobin and 150 μmol/l ABTS in phosphate buffered saline (PBS). In order to start the reaction, 375 μmol/l hydrogen peroxide was added to the mixture, producing a final volume of 1 ml. The solution was immediately vortexed, placed in a 30°C incubator and a clock started. After 165 s, the reaction mixture was transferred to a 1 cm cuvette and absorbance read at 734 nm. Calibrations were performed using the antioxidant Trolox dissolved in PBS, in place of seminal plasma.
Dear Hamza, in our Sci Center we fora long time are dealing with Lipid peroxidation and antioxidant activity in many biological objects, including semen. Published in J. Vestnik of Russian Academy of Med.Sci.( Vestnik Rossiiskoi Academii Medicinskikh nauk), 2015,N1,p.12-16; J. Preclinical Lab. Diagnostiks (Doclinicheskaya laboratornaya diagnostica), 2015,N1, 16-19. and in Russian book @Oxidatvestress and Free Radical Pathologies, Moscow,202, p.37.
Tthe special coefficient of oxidative stress was elaborated and patented. The ssence of it is that no antioxidant activity, no lipid peroxidation products separately can explain the situaton in biological fluids and organism as a whole.
You can contact Center and clarify additional questions witth professor Marina Darenskya (e-mail [email protected])
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