Rupérez F J, Barbas C, Castro M, Martínez S, Herrera E
Facultad de CC Experimentales y Técnicas, Universidad S. Pablo-CEU, Urbanización Montepríncipe, Madrid, Spain.
J Chromatogr A. 1998 Oct 9;823(1-2):483-7. doi: 10.1016/s0021-9673(98)00206-4.
A method for vitamin E (alpha-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 K1 was used as internal standard. Separation was performed by HPLC with methanol-water (96.5:3.5) as eluent in a Nucleosil C18 column (15 x 0.46 cm) at 40 degrees C. 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 K1. Standards and tissue extracts were checked for linearity giving correlation coefficients over 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 95 +/- 3% and 98 +/- 11% 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.
已开发并验证了一种用于测定大鼠脂肪组织和乳腺中维生素E（α-生育酚）的方法。将组织在乙醇-水（1:1）中匀浆，并用正己烷萃取。维生素K1用作内标。在40℃下，以甲醇-水（96.5:3.5）为洗脱剂，在Nucleosil C18柱（15×0.46 cm）上通过高效液相色谱进行分离。维生素E通过荧光检测，激发波长为295 nm，发射波长为350 nm；维生素K1的激发波长为330 nm，发射波长为440 nm。对标准品和组织提取物进行线性检查，脂肪组织中浓度范围为0.56至4.51 nmol/g，乳腺组织中浓度范围为2.18至17.4 nmol/g时，相关系数均超过0.99。批内精密度（相对标准偏差）在3%至4%之间，而批间精密度在8%至9%之间。两种组织的回收率分别在95±3%和98±11%之间。对先前口服过一次该维生素的大鼠进行了维生素E测定。虽然妊娠后期大鼠和未孕大鼠的脂肪组织中维生素E含量没有差异，但妊娠大鼠乳腺中的维生素E含量显著更高，这种差异可能与该组脂蛋白脂肪酶活性增强有关。
