5 mL), and MDA was derivatized with phenylhydrazine (1 [micro]mol; 20 [micro]L of 50 mmol/L aqueous solution).
The obtained results (Table 1) were super-imposable and not significantly different (by Student t-test), confirming that MMDA fulfills the requirements for its use in plasma MDA detection after derivatization with phenylhydrazine, provided that the conditions reported here (pH 4.
Trichloroacetic acid (TCA), ferrous sulfate, glacial acetic acid, sodium dodecyl sulphate (SDS), ethylene diamine tetra acetate (EDTA), sodium nitroprusside, N-(1-naphthyl) ethylenediamine dihydrochloride, phenylhydrazine hydrochloride and Folin Ciocalteu reagent were purchased from SD Fine Chemicals, India.
Assay for phenylhydrazine induced haemolysis of erythrocytes (membrane stabilization study): 20% PCV (packed cell volume) of erythrocyte suspension (from human blood) was prepared according to the procedure described by Hill and Thornalley (1983).
In erythrocyte membrane stabilization study, the extract inhibited the haemolysis of erythrocytes induced by phenylhydrazine in a dose dependent manner, with increased protection from 6.
Phenylhydrazine is one of the autooxidizable substances which, under aerobic conditions may react with molecular oxygen to form a variety of reactive species, e.
indicus root bark, measured in terms of % inhibition of haemolysis induced by phenylhydrazine.
There were three phases in TBARS formation during oxidation of LDL-lipids: (a) a lag phase, during which there was no absorbance and LDL lipids were resistant to oxidation; (b) a propagation phase, characterized by a rapid increase in TBARS (MDA) up to a maximum; and (c) a final phase, which began after ~20 min of incubation with phenylhydrazine, from the point at which the TBARS (MDA) concentration reached its maximum and remained constant for more than 1 h.
001 (a) Capacity in micromoles of MDA formed after incubation with phenylhydrazine, per mole LDL-cholesterol.