Simultaneous determination of vitamins C, E and beta-carotene in human plasma by high-performance liquid chromatography with photodiode-array detection

• Published in: Journal of pharmacy & pharmaceutical sciences Vol. 7; no. 2; pp. 200 - 204
• Main Authors: Zhao, Bin, Tham, Su-Yin, Lu, Jia, Lai, Mui Hoon, Lee, Lionel K H, Moochhala, Shabbir M
• Format: Journal Article
• Language: English
• Published: Canada 30.06.2004
• Subjects: Adult; Antioxidants - analysis; Ascorbic Acid - blood; beta Carotene - blood; Chromatography, High Pressure Liquid - instrumentation; Chromatography, High Pressure Liquid - methods; Humans; Male; Photochemistry - instrumentation; Regression Analysis; Reproducibility of Results; Spectrophotometry, Ultraviolet; Vitamin E - blood
• ISSN: 1482-1826

To develop a high-performance liquid chromatography (HPLC) method with photodiode-array ultraviolet detection for the simultaneous determination of vitamin C, vitamin E and beta-carotene. Following liquid-phase extraction from the human plasma samples, these three vitamins were successfully separated on the LiChrospher 100 RP-18 column (125 x 4 mm I.D.; particle size, 5 microm) at a flow-rate of 1.2 ml/min, with a mobile phase of methanol-acetonitrile-tetrahydrofuran (75: 20: 5, v/v/v). The limits of quantitation were 100, 0.25 and 0.25 microg/ml for vitamin C, vitamin E and beta-carotene respectively. The method is linear over the studied range of 0.25 to 5 g/ml for vitamin E and beta-carotene and 100 to 5000 microg/ml for vitamin C. The extraction recoveries were greater than 83% for these three vitamins. The within day and between-day precision of the analysis did not exceed 15.3 and 16.2%, respectively. A suitable method to determine the concentration of vitamin C, vitamin E and beta-carotene following oral administration of antioxidant supplement capsules to a healthy Chinese volunteer.