Simultaneous determination of retinol, beta-carotene and alpha-tocopherol in adipose tissue by high-performance liquid chromatography.
A method is described for evaluation of fat-soluble vitamin in human adipose tissue with the aim to obtain, accurately and within the shortest analysis time, a time-integrated measure of exposure to vitamins from the diet. Fat tissue was deproteinized with ethanol and extracted with n-hexane. Normal-phase HPLC was performed in a Lichrosorb Si60 column with a gradient of n-hexane-2-propanol at 1 ml/min. Detection was accomplished using a diode-array system (for retinol and beta-carotene) in series with a fluorescence detector (alpha-tocopherol). The method was validated and applied to human adipose tissue in a total of 140 subjects. The mean contents found were 0.43, 0.84, 240.3 microg/g for retinol, beta-carotene and alpha-tocopherol, respectively. The method is sensitive enough for detecting the compounds in 1.6 mg of adipose tissue considering the lowest concentration found.
Casal
S
S
CEQUP/Serviço de Bromatologia, Faculdade de Farmácia do Porto, Portugal. sucasal@ff.up.pt
Macedo
B
B
Oliveira
M B
MB
D000273:Adipose Tissue / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004032:Diet; D006801:Humans; D009203:Myocardial Infarction / Q000453:epidemiology; D011174:Portugal / Q000453:epidemiology; D015203:Reproducibility of Results; D012307:Risk Factors; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D011795:Surveys and Questionnaires; D014801:Vitamin A / Q000032:analysis*; D024502:alpha-Tocopherol / Q000032:analysis*; D019207:beta Carotene / Q000032:analysis*
D019207:beta Carotene; D014801:Vitamin A; D024502:alpha-Tocopherol