Determination of retinol and [alpha]-tocopherol in human seminal plasma using an HPLC with UV detection

Summary Oxidative stress has been proposed as one of the potential causes for infertility in men. Retinol and [alpha]-tocopherol have an important role in the spermatozoa defences against oxidative stress. A method is described here for the simultaneous determination of retinol and [alpha]-tocopherol in human seminal plasma with a suitable sample preparation procedure to prevent retinol and [alpha]-tocopherol degradation. After adequate sample preparation, the samples were determined by reversed-phase column chromatography with UV detection. The analytical performance of this method was satisfactory. The intra-assay and inter-assay coefficients of variation were below 10%. The recoveries were as follows: 90.7% (CV 8.1%) for retinol and 98.2% (CV 4.8%) for [alpha]-tocopherol. No significant differences in both retinol and [alpha]-tocopherol concentration between the smokers and nonsmokers (15 ± 7 nm and 1.86 ± 0.29 µm versus 15 ± 6 nm and 1.93 ± 0.45 µm) were found. A selective high-performance liquid chromatography method for the determination of retinol and [alpha]-tocopherol in human seminal plasma was developed. [PUBLICATION ABSTRACT]