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

Abstract

Oxidative stress has been proposed as one of the potential causes for infertility in men. Retinol and α-tocopherol have an important role in the spermatozoa defences against oxidative stress. A method is described here for the simultaneous determination of retinol and α-tocopherol in human seminal plasma with a suitable sample preparation procedure to prevent retinol and α-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 α-tocopherol. No significant differences in both retinol and α-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 α-tocopherol in human seminal plasma was developed.

Analysis of retinol, α‐tocopherol, 25‐hydroxyvitamin D2 and 25‐hydroxyvitamin D3 in plasma of patients with cardiovascular disease by HPLC–MS/MS method

“…The validated method was successfully applied for the determination of concentrations of retinol, α ‐tocopherol, 25‐OH‐D2 and 25‐OH‐D3 in plasma samples of patients with cardiovascular disease. These concentrations are similar to those reported by other authors for patients with cardiovascular disease (Kanďár, Drábková, Myslíková, & Hampl, ; Robinson‐Cohen et al, ; Wang et al, ) and healthy volunteers (Jenab et al, ; Midttun & Ueland, ; Table ). Several studies showed an association between plasma levels of fat‐soluble antioxidant vitamins and atherosclerotic progression in arteries (Kanďár et al, ; Polidori et al, ; Riccioni et al, ).…”

“…These concentrations are similar to those reported by other authors for patients with cardiovascular disease (Kanďár, Drábková, Myslíková, & Hampl, ; Robinson‐Cohen et al, ; Wang et al, ) and healthy volunteers (Jenab et al, ; Midttun & Ueland, ; Table ). Several studies showed an association between plasma levels of fat‐soluble antioxidant vitamins and atherosclerotic progression in arteries (Kanďár et al, ; Polidori et al, ; Riccioni et al, ). A significant correlation was also confirmed between vitamin D deficiency and incident cardiovascular disease (Judd & Tangpricha, ; Wang et al, ).…”

A review of chromatographic methods for the determination of water‐ and fat‐soluble vitamins in biological fluids

“…methanol) to optimize selectivity . Several methods for the determination of lipovitamins in biological fluids involve chromatographic separation using UV–Vis or fluorescence detection (FLD) . The LODs and LOQs of these methods are usually sufficient for the analysis of carotenoids, retinoids, and tocopherols with appropriate precision and accuracy.…”

“…However, HPLC–FLD methods showed multiple improvements to the sensitivity in comparison with the HPLC–UV methods. For example, LOQ of α‐tocopherol obtained by UV was 0.2 μM , while the value of 21.4 nM was reached by FLD . Measurement of other analytes, such as vitamin K derivatives or vitamin D hydroxy metabolites, is limited because of their low concentrations in the blood.…”

“…For saponification, KOH or isopropanol are used . For further sample purification, LLE is additionally applied using heptane , hexane , ethyl acetate , chloroform , methyl tert ‐butyl ether , or a mixture of hexane/ethanol/acetone/toluene . Deproteinization and LLE resulted in endogenous compounds of the matrix passing into the supernatant, which may affect the separation and determination of the analytes.…”