Simplified method for vitamin E determination in rat adipose tissue and mammary glands by high-performance liquid chromatography.

Abstract

A method for vitamin E (a-tocopherol) measurement in rat adipose tissue and mammary gland has been developed and validated. Tissues were homogenized in ethanol–water (1:1) and extracted with n-hexane. Vitamin K was used as internal 1 standard. Separation was performed by HPLC with methanol–water (96.5:3.5) as eluent in a Nucleosil C column 18 (1530.46 cm) at 408C. Detection was by fluorescence with excitation at 295 nm and emission at 350 nm for vitamin E and at 330 and 440 nm for vitamin K . Standards and tissue extracts were checked for linearity giving correlation coefficients over 1 0.99 in a range of concentrations from 0.56 to 4.51 nmol/g in adipose tissue and from 2.18 to 17.4 nmol/g in mammary gland tissue. Intra-assay precision (R.S.D.) varied between 3 and 4%, whereas inter-assay precision was between 8 and 9%. Recoveries ranged between 9563% and 98611% for the two tissues, respectively. Vitamin E was measured in rats that had previously received one oral dose of this vitamin. Whereas vitamin E content in adipose tissue did not differ between late-pregnant and virgin rats, it was significantly higher in mammary gland of pregnant rats, and this difference could be related to the enhanced lipoprotein lipase activity in this group.