Comparison of a fluorometric assay kit with high-performance liquid chromatography for the assessment of serum retinol concentration

Afr Health Sci. 2015 Jun;15(2):641-6. doi: 10.4314/ahs.v15i2.43.

Abstract

Background: Although high-performance liquid chromatography (HPLC) is the commonly used method for the analysis of retinol in biological samples, simple and rapid test kits are available.

Objectives: This study compared a rapid test kit (ICHECK Fluoro®) to HPLC for the assessment of serum retinol concentrations.

Methods: For the analysis by HPLC, sample preparation included standard deproteinization and extraction phases. The analysis by ICHECK was performed by injecting serum into IEX reagent vials (n=89) and mixing manually for separation. After precipitation of the proteins, the vial was introduced into the chamber of the ICHECK Fluoro and analysed at 0 min (ICHECK0min) and 15 min later (ICHECK15min). Bland and Altman approach was applied to test the agreement between HPLC and ICHECK.

Results: Mean HPLC, ICHECK0min and ICHECK15min values were 421.2±106.0 µg/L, 423.1±118.3 µg/L and 413.2±107.6 µg/L, respectively. Retinol concentrations significantly decreased in the IEX solution over time (p<0.001). No significant proportional bias was observed between HPLC and ICHECK0min (r-0.038, p=0.73) and ICHECK15min (r=-0.024, p=0.82). Fixed biases (HPLC minus ICHECK) for ICHECK0min and ICHECK15min were respectively -1.9±23.1 µg/l (p=0.45) and 8.0±22.7 µg/l (p=0.002).

Conclusion: ICHECK Fluoro may offer a reliable mean for assessing serum retinol for measurements performed with no significant time delay.

Keywords: HPLC; ICHECK Fluoro; serum retinol; test kit; vitamin A status.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Child
  • Chromatography, High Pressure Liquid / methods*
  • Female
  • Fluorometry / instrumentation*
  • Humans
  • Longitudinal Studies
  • Male
  • Morocco
  • Reproducibility of Results
  • Rural Population
  • Sensitivity and Specificity
  • Vitamin A / analysis
  • Vitamin A / blood*
  • Vitamin A Deficiency / blood

Substances

  • Vitamin A