Simultaneous determination of vitamins A and E in rat tissues by high-performance liquid chromatography.
A high-performance liquid chromatographic (HPLC) method to determine vitamins A, Ap and E simultaneously was developed with direct extraction of vitamins from rat tissues with n-hexane and probe sonicating. The dry residue was redissolved in chloroform-methanol. Vitamins A and Ap were detected by UV-Vis and vitamin E by fluorescence. Vitamin K, used as internal standard, was detected both the UV-Vis and by fluorescence. Standards and samples were checked for linearity giving correlation coefficients that were higher than 0.99 in the concentration range of 3.1-9.4 for vitamin A, 8.2-24.7 for vitamin Ap and 0.6-1.7 nmol/g in the case of liver extracts and 0.5-3.0 nmol/g in the case of placenta. The inta-assay precision (R.S.D) varied between 1.48 and 7.25, whereas inter-assay precision was between 4.99 and 7.03. Recoveries ranged between 94 +/- 8 and 107 +/- 5%. Results from the application of this method to different rat tissues having wide range of vitamin content are presented.
Barbas
C
C
Facultad de CC Experimentales y Técnicas, Universidad S. Pablo-CEU, Madrid, Spain.
Castro
M
M
Bonet
B
B
Viana
M
M
Herrera
E
E
D000818:Animals; D001921:Brain / Q000196:embryology; D001923:Brain Chemistry / Q000502:physiology; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002940:Circadian Rhythm; D005260:Female; D005453:Fluorescence; D006586:Hexanes / Q000737:chemistry; D016014:Linear Models; D008099:Liver / Q000737:chemistry / Q000196:embryology; D010920:Placenta / Q000737:chemistry; D011247:Pregnancy; D051381:Rats; D017208:Rats, Wistar; D015203:Reproducibility of Results; D013010:Sonication; D013056:Spectrophotometry, Ultraviolet; D014801:Vitamin A / Q000032:analysis*; D014810:Vitamin E / Q000032:analysis*
D006586:Hexanes; D014801:Vitamin A; D014810:Vitamin E; C026385:n-hexane