A Validated HPLC–DAD Method for Fucoxanthin Determination in Sargassum polycystum C. Agardh

Abstract

Fucoxanthin, an orange pigment found in brown algae, has a distinctive
orange color, making it a natural compound suitable for various
applications, including medical, cosmetic, and food uses. Validation of the
fucoxanthin analysis method is a crucial step to ensure that the quantitative
data obtained are accurate, selective, sensitive, and reproducible, so that
research results can be justified scientifically. The purpose of this study is to
validate the fucoxanthin analytical method using high-performance liquid
chromatography-diode array detector (HPLC-DAD) with criteria such as
selectivity, linearity, accuracy, precision, limit of detection (LOD), and limit
of quantification (LOQ), as well as quantifying the fucoxanthin content of
the Sargassum polycystum C. Agardh fraction. The extraction was
macerated using absolute acetone with a yield of 0.96%. Fractionation was
carried out by TLC using a mixture of n-hexane: acetone (6:4, v/v) as the
eluent. The HPLC system for system suitability testing was realized using a
C18 column stationary phase, a methanol: acetonitrile (70:30) mobile phase,
a flow rate of 0.8mL/ min, a diode array detector, a maximum wavelength
of 447.40nm, and an injection volume of 20µL. The recovery value ranged
from 92.31- 98.11%, linearity with a correlation coefficient (r) of 0.9994,
LOD and LOQ were 7.76 and 25.87ng/ mL, respectively. The brown algae
fraction's fucoxanthin content was found to be 0.83%.