The cytotoxicity of cholesterol and a mixture of beta-sitosterol/campesterol (50%/40%) and their oxides was examined in a cultured-derived macrophage cell line, C57BL/6. Cell numbers, lactate dehydrogenase (LDH) leakage, protein content, lipid uptake, and mitochondria dehydrogenase activity were determined after exposure of cell mononlayers to sterols and sterol oxides at a concentration of 200 microg/mL for up to 120 h. Results indicate that the oxides of cholesterol, beta-sitosterol, and campesterol exhibited similar patterns of toxicity as indicated by LDH leakage, cell viability, and mitochondria dehydrogenase activity. Greatest cell damage was associated with treatments containing 5 alpha,6 alpha-epoxide or cholesterol oxides, followed by beta-sitosterol/campesterol oxides, cholesterol, and beta-sitosterol. The oxides of beta-sitosterol/campesterol caused less LDH leakage and less of an effect on protein content. Results of this study demonstrate that phytosterols contained in vegetable oils, when subjected to frying conditions, do oxidize and may cause cellular damage in an in vitro cell line similar to cholesterol oxides, although less severe.

The oxidative stability of phytosterols in canola, coconut, peanut, and soybean oils was examined under simulated frying conditions of 100, 150, and 180°C for 20 h. The degree of oxidative decomposition was assessed by the loss of phytosterols, accumulation of phytosterol oxides, and the change in fatty acid profiles. The phytosterol oxides produced in the oils were identified using mass spectroscopy. Oils with higher levels of polyunsaturated fatty acids showed greater amounts of sterol loss; however, the sterol loss was less complete than in the more saturated oils. A greater variety of sterol oxides was observed at the lower temperatures of 100 and 150°C compared to 180°C. This study demonstrates that under conditions similar to frying, there is a loss of phytosterols and polyunsaturated fatty acids. The accumulation of phytosterol oxides may be temperature-limited because of further breakdown into products not measurable by typical gas chromatography-mass spectrometry techniques.