Fast Determination of Fat-soluble Vitamins in Pharmaceutical Preparations by High-Performance Liquid Chromatography

OpenAlex İngilizce

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.