Ionic liquid and ultrasound as a fast and innovative combination for improved extraction of Chlorella sorokiniana-derived carotenoids

Chlorella sorokiniana is a microalgae with high amounts of carotenoids, but the extraction methods need to be improved. This work hypothesized that the combination of ionic liquids (ILs) and ultrasound would provide a fast and efficient process for carotenoid extraction, allowing for the reuse of ILs in several extractive cycles. This study aimed to develop an improved ILs/ultrasound-based method for extracting carotenoids from C. sorokiniana. The potential reuse of ILs in five consecutive extraction cycles and the recovery of carotenoids was also examined. Moreover, the antioxidant activity of the carotenoid extracts and their effects on metabolic cell viability and cell death were investigated using HT22 neuronal cells. Initial tests with four ILs and acetone as a control were carried out in an ultrasonic probe. The ILs (1-butyl-3-methylimidazolium chloride [BMIM][Cl]; 1-hexyl-3-methylimidazolium chloride [HMIM][Cl]) were selected for the 2⁵⁻¹ fractional experimental design. The maximized parameters obtained in the experimental design were the following: IL ([HMIM][Cl]), a 1:10 solid-liquid ratio, a 1:4 IL-cosolvent ratio, and two repetitions of extraction with 7.5 min (with an extraction yield of 1.29 mg·g^‐1 of dry matter). A total of 11 carotenoids were separated, and nine were identified, the major ones being lutein and β-carotene. [HMIM][Cl] recyclability using resin Amberlite XAD-7HP was efficient for at least five cycles. On average, 91 % of the IL was recovered, and the pigment yield increased by 40 %. The antioxidant activities of the extracts obtained using [HMIM][Cl] and acetone were 1.65 μmol and 2.32 μmol of α-tocopherol, respectively. Both extracts (≤ 4.0 μg·mL^‐1) exhibited no significant toxicity to HT22 cells. The proposed method is an innovative and improved approach for carotenoid extraction from C. sorokiniana due to its short extraction times and high process yield. [HMIM][Cl] demonstrated stability in reuse cycles and proved to have the potential for obtaining carotenoids from C. sorokiniana.