Has anyone performed assessment of Antioxidant activity in linoleic acid system? i followed this procedure Samples were dissolved in 0.1 M sodium phosphate buffer at pH 7.0, while BHT and BHA were prepared in 50 mM linoleic acid in absolute ethanol. FPH, autolysate or PFS (1 ml) was mixed with 0.5 ml ddH2O and 1 ml 50 mM linoleic acid, after which the mixture was incubated at 60 C in the dark with shaking. For the standard, BHT or BHA solution (1 ml) was added to 1 ml phosphate buffer and 0.5 ml ddH2O, whereas the control was comprised of 1 ml phosphate buffer, 1 ml 50 mM linoleic acid and 0.5 ml ddH2O. The extent of lipid peroxidation was monitored using the ferric thiocyanate method." But my absorbance values don't seem right. If anybody has performed the assay could you please mention how was the sample with linoleic acid prepared? I noticed my linoleic acid wasn't soluble in the system with the above mentioned reagents.
#linoleicacid #antioxidantactivity #proteinhydrolysate #radicalscavenging #lipidoxidation
Yes
A sample solution was prepared by mixing 65 µL of linoleic acid with 5.0 mL sodium phosphate buffer (pH 7) and 2.5 mg of CCEs in 5.0 mL of 75% ethanol. Afterward, 2.5 mL of distilled water was added and the mixture was incubated at 40ºC for 15 minutes. 0.1 mL of sample solution of ferrous chloride (20 mM in 3.5% HCl), 0.1 mL of 30% ammonium thiocyanate and 5.0 mL of 75% ethanol were mixed. The absorbance of this solution at 500 nm was used to determine percentage inhibition using Equation.
I %= (Ac-As/Ac) x 100
Control (treatment without CCE) and sample absorbance values are represented as Ac and As at 3 weeks, respectively and I (%) percentage inhibition. Positive controls included butylated hydroxytoluene at 200 ppm.
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