Evaluation of properties of apigenin and [G-3H]apigenin and analytic method development.

This study provides baseline data and analytical methods to assist in the evaluation of apigenin, a plant flavonoid with promising chemopreventive activity against skin cancer. Apigenin was freely soluble in dimethylsulfoxide (> 100 mg/mL), but it had low solubility (0.00135-1.63 mg/mL) in all the other solvents and surfactants tested, especially in highly hydrophilic or nonpolar solvents. The partition coefficient (log K) calculated from the solubility ratio of apigenin in n-octanol and water was 2.87. Apigenin strongly absorbed UV light, with three maximum absorption wavelengths at 212, 269, and 337 nm (epsilon = 29,800, 19,020, and 18,930 M-1 cm-1, respectively). Using quercetin as the internal standard, a reversed-phase HPLC method was developed to quantitatively analyze apigenin in epidermal cells obtained from SENCAR mice. Apigenin was labeled at position 6, 8, 3', and 5' with tritium by a platinum-catalyzed proton-tritium exchange as confirmed indirectly by 1H NMR analysis of the deuterated apigenin. The tritium label was stable in aqueous environments, especially under acidic and neutral conditions, so [G-3H]apigenin was considered suitable for subsequent absorption and metabolic studies.