Antioxidant activity of sequentially extracted Haematococcus pluvialis using multiple in vitro assays
Aktivitas antioksidan Haematococcus pluvialis yang diekstrak secara berurutan menggunakan beberapa uji in vitro
DOI:
https://doi.org/10.17844/2b3avr58Keywords:
astaxanthin, carotenoids, ethyl acetate, flavonoids, phenolicAbstract
Oxidative stress causes cell damage and various chronic conditions; therefore, natural antioxidants, especially from microalgae, are needed. This study evaluated the antioxidant properties of Haematococcus pluvialis extracts obtained by sequential solvent extraction (n-hexane, ethyl acetate, and methanol) using ultrasonic-assisted maceration. The extracts were characterized for yield, total phenolic content, total flavonoid content, and carotenoid content, and their antioxidant activities were assessed using four in vitro assays (DPPH, ABTS, FRAP, and H₂O₂ scavenging). Sequential extraction yielded cumulative recoveries of 56.13% (30.13% n-hexane, 14.93% ethyl acetate, and 11.07% methanol), exceeding single-solvent methanol extraction (47.13%). The ethyl acetate fraction exhibited the highest phenolic (27.87 ± 0.92 mg GAE/g) and flavonoid (5.96 ± 0.25 mg QE/g) content and contained the highest pigment concentrations (astaxanthin 4,124.25 ± 60.94 µg/mL; total carotenoids 4,686.65 ± 69.25 µg/mL). Consistently, the ethyl acetate extract exhibited the strongest antioxidant activity across all assays (DPPH IC₅₀ = 46.71 µg/mL; ABTS IC₅₀ = 35.85 µg/mL; FRAP IC₅₀ = 23.94 µg/mL; H₂O₂ IC₅₀ = 54.64 µg/mL), outperforming the other sequential fractions and the single-solvent methanol extract. These results indicate that semi-polar extraction with ethyl acetate effectively concentrates phenolic, flavonoid, and pigment antioxidants from H. pluvialis. Sequential extraction enhances recovery and facilitates the identification of the most bioactive fraction, supporting the potential use of ethyl acetate extracts as natural antioxidants for nutraceutical, cosmetic, and pharmaceutical applications.
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