Abstract
A precise, accurate and rapid HPLC-UV method for simultaneous determination of fat-soluble vitamins (vitamin D3, E-acetate, K1, β-carotene, A-palmitate) and coenzyme Q10 was developed and validated according to ICH guidelines. Optimal chromatographic separation of the analytes in minimal analysis time (8 min) was achieved on a Luna C18 150 × 4.6 mm column using a mixture of acetonitrile, tetrahydrofuran and water (50:45:5, v/v/v). The described reversed phase HPLC method is the first published for quantification of these five fat-soluble vitamins and coenzyme Q10 within a single chromatographic run. The method was further applied for quantification of the analytes in selected liquid and solid dosage forms, registered as nutritional supplements and prescription medicines, which confirmed its suitability for routine analysis.
Highlights
Fat-soluble vitamins (FSVs) are essential for a variety of biochemical and physiological functions in human body, such as epithelial cell differentiation and vision, calcium and phosphate homeostasis, antioxidative protection in cell membranes and blood coagulation.[1,2] As FSV, with the exception of vitamin D3, cannot be obtained by endogenous synthesis, food or vitamin preparations are their main sources
We aimed to develop rapid, accurate, precise and selective HPLC-UV method for simultaneous quantification of FSVs and coenzyme Q10, using simple and fast extraction procedures
THF and water at flow rates 0.5–2.0 mL/min and column temperatures 25–40 °C and detection wavelengths 210–325 nm were tested to optimize the chromatographic separation of all tested vitamins as well as both reduced and oxidized form of Coenzyme Q10 (CoQ10)
Summary
Fat-soluble vitamins (FSVs) are essential for a variety of biochemical and physiological functions in human body, such as epithelial cell differentiation and vision (vitamin A), calcium and phosphate homeostasis (vitamin D), antioxidative protection in cell membranes (vitamin E) and blood coagulation (vitamin K).[1,2] As FSV, with the exception of vitamin D3, cannot be obtained by endogenous synthesis, food or vitamin preparations are their main sources. Due to the large number of commercially available vitamin preparations and their widespread use, the quality control of these preparations is extremely important, especially because of the risk of toxicity from excessive intake of vitamins A and D.3. The published HPLC-UV methods for quality control of preparations containing FSVs are quite limited in terms of separation and simultaneous determination of FSVs,[4,5,7,8,11] often have run time longer than 15 min[4,5,6,9,12] and generally require complicated and time-consuming sample preparation.[11,12,15,16] Further on, to our knowledge none of the published HPLC methods offer simultaneous determination of FSVs and β-carotene and coenzyme Q10, which are often found along in medicines and nutritional supplements
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