Background:
Retinol and vitamin E are fat-soluble vitamins crucial for human health, yet
their isomers’ distributions in the human body are still known roughly. In order to figure out the physical
condition and evaluate the nutritional status of an individual, it is imperative to analyze retinol and
VE isomers in human serum.
Objective:
This work aims to establish a rapid and simple high-performance liquid chromatography
with fluorescence detection for simultaneous determination of retinol and vitamin E isomers in human
serum.
Methods:
Separation was accomplished on a common C18 column thermostated at 25 oC, using a simple
isocratic elution program of methanol/acetonitrile (85:15, v/v) at a flow rate of 1.0 mL/min. Fluorescence
detection was operated using excitation/emission wavelengths of 329 nm/472 nm for retinol and
294 nm/338 nm for VE isomers, respectively.
Results:
Rapid separation was achieved within 13 min. Linear ranges of the method were 0.020-50.0
µg/mL, with correlation coefficients greater than 0.999. Detection limits and the quantification limits
were 0.001-0.004 µg/mL and 0.003- 0.013 µg/mL, respectively. Mean recoveries were 84.1%- 98.2%,
with intra-day and inter-day relative standard deviations less than 12.3% and 13.6%, respectively. This
method has been applied to the simultaneous determination of retinol and 8 VE isomers in human serum
samples with satisfactory results.
Conclusion:
A rapid, simple and robust method was developed for routine analysis of retinol and eight
vitamin E isomers in human serum, providing a useful tool for clinical diagnosis and nutritional evaluation.
