Determination of β-Carotene in Spirulina by ASE-UPLC
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Abstract:
A novel method has been developed for the rapid extraction and determination of β-Carotene in Spirulina by accelerated solvent extraction (ASE) coupled with ultra performance liquid chromatography (UPLC). The ASE parameters (extraction solvent, temperature, static time and cycles) were optimized by Single factor experiment and Orthogonal experiment design in order to maximize the extraction efficiency. The result showed that the optimum condition for extraction was as followed: petroleum ether (30~60 ℃) as solvent, extraction temperature of 120 ℃, extraction time of 5 min and static loop of three times. The ASE extract was then diluted and determined by UPLC on the Agilent Eclipse Pluse C18 column (1.8 μm, 2.1 mm×50 mm) at 25 ℃, using 85% CH3OH-15% CH3CN as mobile phase and a Diode array detector at 450 nm. The results showed that the linear ranges of β-Carotene was 1~50 μg/mL with correlation coefficient higher than 0.999. The recoveries ranged from 98.65% to 103.18% with RSDs of 0.68~1.74% and the limit of detection was 0.10 μg/kg. The method was simple, rapid and accurate, with high sensitivity and reproducibility, which was suitable for the determination of β-Carotene in Spirulina.
