2.11. Lipid Peroxidation Analysis

The thiobarbituric acid-reactive substances (TBARS) assay was used to assess oxidative stress, specifically lipid peroxidation, by reacting the secondary end product of the oxidation of polyunsaturated fatty acids (malondialdehyde) with thiobarbituric acid (TBA) in a colorimetric reaction to form TBARS [56,57]. Briefly, SH-SY5Y cells were grown in 6-well plates until confluent. Following pre-treatment with the relevant free or nanoformulated conditions and 24 h rotenone treatment, cells were washed once with DPBS and lysed at 4 °C as described above. The TBARS assay was carried out in accordance with manufacturer guidelines (R&D Systems, Parameter TM) and as previously described [42]. Freshly prepared TBA was added to TBARS acid-treated cell lysate which was incubated at 60 °C for 2.5 h. The absorbance of samples was read at 532 nm before and after incubation to estimate the formation of TBARS.