Cholesterol as a singlet oxygen detector in biological systems

In cells under oxidative attack, membrane Ch, through the formation of its signature hydroperoxide and diol products, can serve as a unique detector in situ, allowing discrimination between 1O2 and free radical intermediacy. Of the two techniques described for analyzing Ch oxidation products, TLC with color development suffices for preliminary, mainly qualitative product screening, whereas a high-performance approach such as HPLC-EC(Hg) is advised when maximum resolution and sensitivity of quantitation are necessary. By using these strategies, one can monitor the formation of 1O2, for example, in a biologically relevant milieu (membrane), thus avoiding the difficulties associated with external detection, e.g., by physical means. These approaches would be valuable for assessing reaction mechanisms for various oxidative agents of biomedical importance, including environmental phototoxins and the rapidly emerging family of phototherapeutic drugs. Although photodynamic stress has been emphasized, the methods described should have broad applicability in the elucidation of oxidative mechanisms.