Light Increases Astaxanthin Absorbance in Acetone Solution through Isomerization Reactions.

Abstract

Astaxanthin quantitative analysis is prone to high variability between laboratories. This study aimed to assess the effect of light on the spectrometric and high-performance liquid chromatography (HPLC) measurements of astaxanthin. The experiment was performed on four Haematococcus pluvialis-derived astaxanthin-rich oleoresin samples with different carotenoid matrices that were analyzed by UV/Vis spectrometry and HPLC according to the United States Pharmacopoeia (USP) monograph. Each sample was dissolved in acetone in three types of flasks: amber glass wrapped with aluminium foil, uncovered amber glass, and transparent glass. Thus, the acetone solutions were either in light-proof flasks or exposed to ambient light. The measurements were taken within four hours (spectrometry) or three hours (HPLC) from the moment of oleoresin dissolution in acetone to investigate the dynamics of changes in the recorded values. The results confirm the logarithmic growth of astaxanthin absorbance by 8-11% (UV/Vis) and 7-17% (HPLC) after 3 h of light exposure. The changes were different in the samples with different carotenoid matrices; for instance, light had the least effect on the USP reference standard sample. The increase in absorbance was accompanied with the change of isomeric distribution, namely a reduction of 13Z and an increase of All-E and 9Z astaxanthin. The greater HPLC values' elevation was related not only to the increase of astaxanthin absorbance, but also to light-dependent degradation of internal standard apocarotenal. The findings confirm a poor robustness of the conventional analytical procedure for astaxanthin quantitation and a necessity for method revision and harmonization to improve its reproducibility.