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Ultrasound-Assisted Enzymatic Extraction of Anthocyanins from Raspberry Wine Residues: Process Optimization, Isolation, Purification, and Bioactivity Determination

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Abstract

Raspberry wine residues (RWR) contain abundant anthocyanins. The extraction conditions for the ultrasound-assisted enzymatic extraction (UAEE) of anthocyanins from RWR were optimized using response surface methodology coupled with genetic algorithm. The optimum extraction parameters were extraction temperature of 44 °C, ultrasound power of 290 W, pectinase dosage of 0.16%, and extraction time of 30 min. Results showed that UAEE exhibited the highest anthocyanin yield (0.853 ± 0.009) mg/g and extraction efficiency (84.75 ± 2.11)% when compared with the other three conventional extraction methods (hot water-, acidified ethanol-, and enzymatic-assisted extractions). UAEE method was available for realizing high activities of DPPH (417.15 ± 11.83) Trolox equivalents/g extract and ABTS (520.07 ± 8.29) Trolox equivalents/g extract, and reducing power (412.79 ± 9.11) Trolox equivalents/g extract. AB-8 macroporous resin, Sephadex LH-20 column chromatography, and high-speed countercurrent chromatography (HSCCC) were used to separate and purify anthocyanins in turn. The main anthocyanins were identified by using HPLC-MS/MS and NMR. The two main anthocyanins (cyanidin-3-glucoside with the purity of 93.46% and cyanidin-3-rutinoside with the purity of 94.16%) were obtained from RWR. Ultimately, we further evaluated the anti-tumor activity of the main anthocyanins on lung cancer. Results showed that the anti-tumor effect of cyanidin-3-rutinoside on A549 cells was better than that of cyanidin-3-glucoside. In addition, cyanidin-3-rutinoside could memorably increase intracellular ROS levels and apoptosis of MCF-7 cells. The UAEE is an effective and rapid method. This method has a broad application prospect in the extraction of bioactive substances from natural plant resources.

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Acknowledgements

The authors would like to thank the Key Laboratory of Particle & Radiation Imaging, Ministry of Education.

Funding

This study was funded by the National Science and Technology Support of China (20161850240).

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Correspondence to Xu Cai.

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This article does not contain any studies with human participants or animals performed by any of the authors. In this experiment, we did not collect any samples of human and animals. The MCF-7 cells were used in this paper, which was purchased from Concorde cell bank with national standard (Beijing, China).

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There is no conflict of interest among all the authors (Hongkun Xue, Jiaqi Tan, Qian Li, Jintian Tang, Xu Cai) of this paper.

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Xue, H., Tan, J., Li, Q. et al. Ultrasound-Assisted Enzymatic Extraction of Anthocyanins from Raspberry Wine Residues: Process Optimization, Isolation, Purification, and Bioactivity Determination. Food Anal. Methods 14, 1369–1386 (2021). https://doi.org/10.1007/s12161-021-01976-8

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