High precision, good recovery and fast determination of alfa-tocopherol (Vitamin E) in serum and plasma samples? This is your analytical method

Abstract

Vitamin E comprises a family of four tocopherols and four tocotrienols (α, β-, γ- and δ-), all of them antioxidants compounds. α-Tocopherol (α-T) showed superior biological activity [1] over the other compounds and is the major tocopherol present in blood and tissues [2]. Being antioxidant α-T reduce oxidative stress (OS) and could be implicated in the etiopathology of a number of diseases as reducing the risk of cancer, cardiovascular and neurodegenerative diseases [3]. This experimental work have been focused to develop a high quality, specific and fast analytical methodology for determination of α-T in biological samples using LC-ESI-MS. For that, chromatographic (mobile phase composition and flow, temperature and column C-18 2.1 x 50 mm-1.8 μm), electrospray ionization (negative) and mass spectrometric parameters (quali and quanti transitions, gas flow and temperature, nebulizer) have been carefully optimized. In addition, validation procedure (precision, recoveries, limits of quantitation, linearity…) have been carried out using a deutered compound of α-T (d6-α-T) in order to obtain a reproducible, sensible and accuracy analytical method. The optimized and validated analytical method has been applied for determination of concentration of α-T in serum and plasma samples. The pretreatment of biological samples from total blood have been also optimized.