Simultaneous determination of retinol, retinyl palmitate and β-carotene in rat serum treated with 7,12-dimethylbenz[a]anthracene and Hypericum Perforatum L. by high-performance liquid chromatography with diode-array detection

Abstract

AbstractA new and simple high-performance liquid chromatography method was developed and validated for the simultaneous determination of retinol, retinyl palmitate and β-carotene in rat serum treated with Hypericum Perforatum L. and 7,12-dimethylbenz[a]anthracene. Furthermore, vitamin C was determined spectrophotometrically. High-performance liquid chromatography analysis was performed utilizing an Inertsil ODS3 reversed phase column with methanol-acetonitrile-tetrahydrofuran (65:30:5, v/v/v) as mobile phase, at a flow rate of 1.5 mL min−1 and 40°C. Diode-array detection was conducted at 325 and 450 nm for retinol and retinyl palmitate, and β-carotene, respectively with a running time of 26 min. The high-performance liquid chromatography assay and extraction procedure proposed are simple, rapid, sensitive and accurate. This method was then applied to determine the amounts of retinol, retinyl palmitate and β-carotene in rat serum. Results of this study demonstrated that at 60th day in the 7,12-dimethylbenz[a]anthracene-treated group there was a significant decrease (pa] anthracene + Hypericum Perforatum L. treated group compared to the control group..