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We developed a selective bioanalytical RP-HPLC method for estimation of vitamin E from dried blood spot (DBS) sample, a potential technique which can be used for population-based epidemiological studies. Vitamin E was extracted from DBS by using liquid-liquid extraction technique with methanol (100% v/v) as reconstituting solvent for the residue. Alpha tocopheryl acetate was used as internal standard. Samples were analyzed directly on HPLC with C18 (250 × 4.6 mm × 5 μm) Phenomenex column. The mobile phase used was methanol to water (99:1% v/v) at a flow rate of 1.4 mL/min. The detector wavelength used was 292 nm. The retention time observed for vitamin E and internal standard was 10.225 ± 0.00075 min and 13.580 ± 0.00075 min respectively. The vitamin E calibration curve was found to be linear over the range of 0.625 to 60 μg/mL. The limit of quantification for vitamin E was found to be 0.1 μg/mL. Accuracy of the developed method was found to be 103.179%, 101.625%, and 100.174% with percentage of coefficient of variation of 0.0161, 0.0215, and 0.2790 for HQC, MQC, and LQC samples respectively which were within USFDA acceptance limit of ± 15 to ± 20%.The intraday and interday precision expressed as coefficient of variation were 0.0191–0.0841% and 0.0074–0.0252% respectively. The method represents a simple, rapid, specific, accurate, and precise method for estimation of vitamin E in human blood using DBS technique. The developed method can be further evaluated with respect to effect of matrix variability before it can be used in clinical setting.

Chromatographic method for estimation of vitamin E from dried blood spot sample