Abstract
Background:
There are limited evidence based studies demonstrating the stability of fat-soluble vitamins (FSV) measured in blood. This study aimed to examine the effects of light, temperature and time on vitamins A, D and E throughout the total testing process.
Methods:
Four experiments were conducted. Three investigated the sample matrix, of whole blood, serum and the extracted sample, against the variables of temperature and light; and the fourth experiment investigated the sample during the extraction process against the variable of light. All samples were analysed via our simultaneous FSV method using liquid chromatography-tandem mass spectrometry technology. The allowable clinical percentage change was calculated based on biological variation and desirable method imprecision for each analyte. The total change limit was ±7.3% for 25-OH-vitamin D3, ±11.8% for retinol and ±10.8% for α-tocopherol.
Results:
Vitamins D and E were stable in the investigated conditions (concentration changes <4%) in the pre-analytical and analytical stages. Vitamin A showed photosensitivity in times >48 h with concentration changes of −6.8% (blood) and −6.5% (serum), both are within the allowable clinical percentage change. By contrast, the extracted retinol sample demonstrated a concentration change of −18.4% after 48 h of light exposure. However, vitamin A in the serum and extracted solution was stable for one month when stored at −20°C.
Conclusions:
Blood samples for vitamins D and E analyses can be processed in normal laboratory conditions of lighting and temperature. The required conditions for vitamin A analysis are similar when performed within 48 h. For longer-term storage, serum and vitamin A extracts should be stored at −20°C.
Acknowledgments:
The work performed at RMIT University was conducted in the RMIT-Agilent Clinical Biochemistry Mass Spectrometry Collaboration Laboratory. We wish to thank Mrs Annabel Mitchell and Dr. Chris Fouracre from Agilent Technologies for their input into the method development.
Author contributions: All authors contributed equally to the development and analysis of this study. Dr. Ali Albahrani conducted the experiments in the clinical biochemistry laboratories, RMIT University, and he wrote the first draft of the manuscript. All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: This work was part of Dr. Ali A. Albahrani’s PhD project which was supported in kind by Agilent Technologies as part of the RMIT Clinical Biochemistry – Agilent Mass Spectrometry Collaboration.
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organisation(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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