Title:Fast Determination of Fat-soluble Vitamins in Pharmaceutical Preparations by High-Performance Liquid Chromatography
Volume: 21
Issue: 2
Author(s): Gamze Ergin Kızılçay*, Sena Çağlar Andaç and Sıdıka Ertürk Toker
Affiliation:
- Department of Analytical Chemistry, Faculty of Pharmacy, Istanbul University, Istanbul, 34116, Turkey
Keywords:
Fat-soluble vitamins, simultaneous determination, HPLC, vitamin A, vitamin D3, vitamin E and K1 vitamins.
Abstract:
Background: Vitamins are needed for the healthy functioning of the body. When these
vitamins are not in sufficient amounts in the body, they are usually taken externally with
pharmaceutical preparations. Taking vitamins into the body in the right amounts is possible by
analyzing the amounts in pharmaceutical preparations with validated methods.
Methods: In this study, a fast, simple, easily applicable, and selective normal-phase HPLC method
was developed for the simultaneous determination of fat-soluble A, D3, E, and K1 vitamins in
pharmaceutical preparations from syrup and tablets. Separation of the vitamins was carried out on a
Zorbax CN column (250 x 4.6 mm, 5 µm) using a mixture of hexane-isopropyl alcohol (98:2, v/v) at
30°C column temperature and 1 ml/min flow rate. The detection wavelength is 280 nm. The
developed method has been validated according to ICH Harmonised Tripartite Guideline Validation
of Analytical Procedures: Text and Methodology Q2(R1) rules.
Results: Calibration graphs are linear over the range of 10-1000 μg ml-1, 1-50 μg ml-1, 10-2000 μg
ml-1, 0.5-20 μg ml-1, and the limit of detection values were found to be 1.496, 0.280, 1.388 and 0.040
μg ml-1 for A, D3, E and K1 vitamins, respectively. Relative standard deviation values, which express
within-day and between-day repeatability, were found below 2.54%. Average recovery values were
also found at about 100.28, 101.46, 100.65, and 100.29% for A, D3, E and K1 vitamins, respectively.
Conclusion: The developed and validated method was successfully applied to the simultaneous
analysis of fat-soluble vitamins A, D3, E and K1 in pharmaceutical preparations in syrup and tablet
form.
