Evaluation of 25-OH vitamin D by high performance liquid chromatography: validation and comparison with electrochemiluminescence

Keyfi, F. and Nahid, S. and Mokhtariye, A. and Nayerabadi, S. and Alaei, A. and Varasteh, A. R. (2018) Evaluation of 25-OH vitamin D by high performance liquid chromatography: validation and comparison with electrochemiluminescence. Journal of Analytical Science and Technology, 9 (1).

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Abstract

Objectives: The aim of the present study was to develop a robust and easy to use high performance liquid chromatography (HPLC) to analyze 25(OH)D3 in human serum. Background: Vitamin D is a fat-soluble steroid hormone precursor that is mainly produced in the skin by exposure to sunlight. It is also supplied in the diet and plays a pivotal role in calcium homeostasis and skeletal metabolism throughout life. Methods: To assess its analytical performance, we used the RECIPE HPLC Complete Kit and an HPLC-UV instrument. Our HPLC results were compared with a validated electrochemiluminescence method. Results: The method was linear for the lower limit of quantification from 3 ng/l up to at least 200 ng/l for 25(OH)D3, with the following equation for the regression line: y = 0.172 X + 2.45 (R2 = 0.989). Intra-assay precision was determined by extracting and quantifying 10 serum replicates from one patient. The mean was 37.875 ng/ml, the standard deviation was 0.22, and the coefficient of variation was 0.58. Comparisons of results demonstrated good agreement between HPLC and ECL methods (R2 = 0.883). Conclusion: The HPLC assay demonstrates excellent linearity, acceptable accuracy and precision, and good agreement with a validated ECL method. The simple sample preparation and ease of use make it practical for the routine clinical laboratory. © 2018, The Author(s).

Item Type: Article
Additional Information: Cited By :1 Export Date: 16 February 2020 Correspondence Address: Varasteh, A.-R.; Immunobiochemistry Lab, Allergy Research Center, Mashhad University of Medical SciencesIran; email: varasteha@varastegan.ac.ir
Uncontrolled Keywords: 25-OH vitamin D Electrochemiluminescence High-performance liquid chromatography Validation
Subjects: QU Biochemistry
QW Microbiology and Immunology
Divisions: Mashhad University of Medical Sciences
Depositing User: lib2 lib2 lib2
Date Deposited: 12 May 2020 03:00
Last Modified: 12 May 2020 03:00
URI: http://eprints.mums.ac.ir/id/eprint/17364

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