Abstract
BackgroundWe 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.ResultsThe 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.ConclusionsThe 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.
Highlights
We developed a selective bioanalytical RP-high-performance liquid chromatography (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
Detection wavelength of 292 nm was selected after scanning the standard solution of vitamin E over the range of 200–400 nm
System suitability For system suitability test, the mixture solution of 60 μg/ mL vitamin E and 30 μg/mL internal standard was prepared. Parameters such as theoretical plate count, tailing factors, resolution, and reproducibility in retention time (RT) for six repetitions of vitamin E and internal standard were explored for compliance with the system suitability test
Summary
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. Oxidative stress in lipid component of cell leads to significant disturbance of cell homeostasis. Such stress has been identified as contributory factor in pathogenesis of various chronic diseases like, diabetes mellitus [1], Parkinson’s disease [2], cardiovascular diseases [3], etc. In spite of its widespread physiological importance and probable involvement in several chronic diseases, only few epidemiological studies are available for assessing role of vitamin E. A major hindrance in conducting such studies is the lack of availability of simple and cheap techniques for sample collection, preservation, and analysis
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