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


METHODS
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).


RESULTS
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.


CONCLUSIONS
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.
Zhao
Bin
B
Defence Medical and Environmental Research Institute, DSO National Laboratories, (DMERI @ DSO), Singapore, Republic of Singapore.
Tham
Su-Yin
SY
Lu
Jia
J
Lai
Mui Hoon
MH
Lee
Lionel K H
LK
Moochhala
Shabbir M
SM
D000328:Adult; D000975:Antioxidants / Q000032:analysis*; D001205:Ascorbic Acid / Q000097:blood*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods; D006801:Humans; D008297:Male; D010777:Photochemistry / Q000295:instrumentation; D012044:Regression Analysis; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D014810:Vitamin E / Q000097:blood*; D019207:beta Carotene / Q000097:blood*
D000975:Antioxidants; D019207:beta Carotene; D014810:Vitamin E; D001205:Ascorbic Acid