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A Rapid and Sensitive HPLC-FLD Method for the Determination of Retinol and Vitamin E Isomers in Human Serum

Author(s):

Yixia Yang, Dan Lu, Danni Yang, Shuo Yin, Jing Zhang, Bo Zheng, Yongxin Li and Chengjun Sun*   Pages 1 - 6 ( 6 )

Abstract:


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. Method: 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

Keywords:

High-performance liquid chromatography (HPLC), fluorescence detection (FLD), retinol, tocopherol, tocotrienol, human serum.

Affiliation:

Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Xi`an Jiaotong University Health Science Center, School of Public Health, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital, Sichuan University, West China School of Public Health, No. 4 West China Teaching Hospital



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