Various beneficial properties has been attributed to nigella sativa, including its antioxidant potential. Previously, it was reported that supercritical fluid extraction (SFE) could be used to obtain N. sativa extract rich in antioxidants. In the present study, N. sativa extracts prepared using the previously optimized SFE as well as the traditional Soxhlet extraction approaches were analyzed for various known antioxidants. N. sativa extracts were found to prevent protein carbonyl formation as well as depletion of intracellular glutathione (GSH) in fibroblasts exposed to toluene. Furthermore, partially purified SFE and Soxhlet fractions could prevent loss of hepatic GSH in toluene-induced oxidative stressed Wistar rats as well as in L929 fibroblasts. The results showed that SFE-produced N. sativa extract is richer in antioxidants than the Soxhlet approach. It was also shown using preparative silica gel and reverse phase chromatography that different fractions of SFE-extracted or Soxhlet-extracted N. sativa had different levels of protective effects with regards to GSH depletion in vivo as well as in cell culture. Although fractions rich in thymoquinone were found to be most potent in terms of antioxidant capacity, the data indicates that the protective effects of N. sativa may not only be due to thymoquinone, but perhaps other antioxidants.
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