{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T23:58:49Z","timestamp":1780531129431,"version":"3.54.1"},"reference-count":37,"publisher":"MDPI AG","issue":"13","license":[{"start":{"date-parts":[[2022,7,4]],"date-time":"2022-07-04T00:00:00Z","timestamp":1656892800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"the National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["32171862"],"award-info":[{"award-number":["32171862"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"the National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["SXLk2020-0201"],"award-info":[{"award-number":["SXLk2020-0201"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"the National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["2021NY-067"],"award-info":[{"award-number":["2021NY-067"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"the Shaanxi Academy of Forestry Science and Technology innovation plan special","award":["32171862"],"award-info":[{"award-number":["32171862"]}]},{"name":"the Shaanxi Academy of Forestry Science and Technology innovation plan special","award":["SXLk2020-0201"],"award-info":[{"award-number":["SXLk2020-0201"]}]},{"name":"the Shaanxi Academy of Forestry Science and Technology innovation plan special","award":["2021NY-067"],"award-info":[{"award-number":["2021NY-067"]}]},{"name":"the Research and Development Projects in Shaanxi Province","award":["32171862"],"award-info":[{"award-number":["32171862"]}]},{"name":"the Research and Development Projects in Shaanxi Province","award":["SXLk2020-0201"],"award-info":[{"award-number":["SXLk2020-0201"]}]},{"name":"the Research and Development Projects in Shaanxi Province","award":["2021NY-067"],"award-info":[{"award-number":["2021NY-067"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Red Malus \u2018Royalty\u2019 fruits are rich in anthocyanins. This study aimed to obtain the optimal parameters for the extraction and separation of anthocyanins from Malus \u2018Royalty\u2019 fruits and to evaluate the inhibitory effect of the enriched anthocyanin fraction on gastric cancer cells. Ultrasonic-assisted extraction was used for the extraction of the anthocyanins of Malus \u2018Royalty\u2019 fruit, and the extraction results showed that the optimum parameters were an extraction temperature of 20 \u00b0C, a solid\u2013liquid ratio of 1:6 (g\/mL), ethanol and formic acid contents of 70% and 0.4%, respectively, an extraction time of 40 min, and an ultrasonic power of 300 W. The optimum extraction parameters to achieve the highest anthocyanin yield by a single-factor experiment coupled with response surface methodology were identified. The separation results showed that the AB-8 macroporous resin was a better purifying material, with 60% ethanol as an adsorbent, and the adsorption\u2013desorption equilibrium times were 6 h and 1 h, respectively. Cyanidin-3-galactoside was the main body composition separation of anthocyanins by a high-performance liquid chromatography-diode array detector. The antitumor activity results showed that the anthocyanins of Malus \u2018Royalty\u2019 fruits have a significant inhibitory effect on the gastric cancer cell line BGC-803. The in vitro cell viability test of CCK-8 showed that the inhibitory effect on tumor cells was more significant with the increased anthocyanin concentration, with a half maximal inhibitory concentration (IC50) value of 105.5 \u03bcg\/mL. The cell morphology was observed by an inverted microscope, and it was found that the backbone of BGC-803 treated with a high concentration of anthocyanins was disintegrated and the nucleoplasm was concentrated. The mechanism of apoptosis was analyzed by Western blotting, and the results showed that with increasing anthocyanin concentration in the medium, the expression levels of the proapoptotic proteins Bax and Bak increased, and the expression levels of the antiapoptotic proteins Bcl-2 and Bcl-xL decreased, which coordinated the regulation of cell apoptosis. This research suggests that the enriched anthocyanin fraction from Malus \u2018Royalty\u2019 fruits have potential antitumor and adjuvant therapeutic effects on gastric cancer.<\/jats:p>","DOI":"10.3390\/molecules27134299","type":"journal-article","created":{"date-parts":[[2022,7,4]],"date-time":"2022-07-04T20:59:18Z","timestamp":1656968358000},"page":"4299","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Ultrasound-Assisted Extraction of Anthocyanins from Malus \u2018Royalty\u2019 Fruits: Optimization, Separation, and Antitumor Activity"],"prefix":"10.3390","volume":"27","author":[{"given":"Yixin","family":"Liu","sequence":"first","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuheng","family":"Zhao","sequence":"additional","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yue","family":"Zhuo","sequence":"additional","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuwen","family":"Li","sequence":"additional","affiliation":[{"name":"Shanghai United International School, Gubei Secondary Campus, Shanghai 201103, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiaxin","family":"Meng","sequence":"additional","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yilin","family":"Wang","sequence":"additional","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Houhua","family":"Li","sequence":"additional","affiliation":[{"name":"College of Landscape Architecture and Art, Northwest A & F University, Yangling 712100, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,7,4]]},"reference":[{"key":"ref_1","first-page":"1649","article-title":"SHCBP1 Is a Novel Target and Exhibits Tumor-Promoting Effects in Gastric Cancer","volume":"41","author":"Dong","year":"2018","journal-title":"Oncol. Rep."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"E359","DOI":"10.1002\/ijc.29210","article-title":"Cancer Incidence and Mortality Worldwide: Sources, Methods and Major Patterns in GLOBOCAN 2012","volume":"136","author":"Ferlay","year":"2015","journal-title":"Int. J. Cancer"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1296","DOI":"10.2174\/1389450118666170123145357","article-title":"The Antioxidant Activities of Natural Polysaccharides","volume":"18","author":"Huang","year":"2017","journal-title":"Curr. Drug Targets"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"105321","DOI":"10.1016\/j.phrs.2020.105321","article-title":"A Review on the Relationship of Mast Cells and Macrophages in Breast Cancer\u2014Can Herbs or Natural Products Facilitate Their Anti-Tumor Effects?","volume":"164","author":"Gou","year":"2021","journal-title":"Pharmacol. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"e13028","DOI":"10.1111\/jfbc.13028","article-title":"Phenolic Compounds Present in Fruit Extracts of Malus Spp. Show Antioxidative and Pro-Apoptotic Effects on Human Gastric Cancer Cell Lines","volume":"43","author":"Han","year":"2019","journal-title":"J. Food Biochem."},{"key":"ref_6","unstructured":"Li, H.H., and Zhao, B. (2014). Chinese Crabapple, Chinese Forestry Publishing House."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Liu, J., Zhou, H., Song, L., Yang, Z., Qiu, M., Wang, J., and Shi, S. (2021). Anthocyanins: Promising Natural Products with Diverse Pharmacological Activities. Molecules, 26.","DOI":"10.3390\/molecules26133807"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1361779","DOI":"10.1080\/16546628.2017.1361779","article-title":"Anthocyanidins and Anthocyanins: Colored Pigments as Food, Pharmaceutical Ingredients, and the Potential Health Benefits","volume":"61","author":"Khoo","year":"2017","journal-title":"Food Nutr. Res."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"509","DOI":"10.1016\/j.ultsonch.2015.06.021","article-title":"Effect of Ultrasound on the Extraction of Total Anthocyanins from Purple Majesty Potato","volume":"27","author":"Mane","year":"2015","journal-title":"Ultrason. Sonochem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.foodchem.2015.02.045","article-title":"Extraction of Anthocyanins and Polyphenols from Black Rice (Oryza Sativa L.) by Modeling and Assessing Their Reversibility and Stability","volume":"191","author":"Pedro","year":"2016","journal-title":"Food Chem."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"3072","DOI":"10.1080\/10408398.2015.1087963","article-title":"Anthocyanin Extraction from Plant Tissues: A Review","volume":"57","author":"Silva","year":"2017","journal-title":"Crit. Rev. Food Sci. Nutr."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Gecchele, E., Negri, S., Cauzzi, A., Cuccurullo, A., Commisso, M., Patrucco, A., Anceschi, A., Zaffani, G., and Avesani, L. (2021). Optimization of a Sustainable Protocol for the Extraction of Anthocyanins as Textile Dyes from Plant Materials. Molecules, 26.","DOI":"10.3390\/molecules26226775"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"124993","DOI":"10.1016\/j.foodchem.2019.124993","article-title":"Optimization Model for Ultrasonic-Assisted and Scale-up Extraction of Anthocyanins from Pyrus Communis \u201cStarkrimson\u201d Fruit Peel","volume":"297","author":"Belwal","year":"2019","journal-title":"Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"10410","DOI":"10.1038\/s41598-020-67266-1","article-title":"Optimization of Ultrasonic-Assisted Extraction of Polysaccharides from Purple Glutinous Rice Bran (Oryza Sativa L.) and Their Antioxidant Activities","volume":"10","author":"Surin","year":"2020","journal-title":"Sci. Rep."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"3731","DOI":"10.1111\/1750-3841.15497","article-title":"Ultrasound-Assisted Enzymatic Extraction of Anthocyanins from Grape Skins: Optimization, Identification, and Antitumor Activity","volume":"85","author":"Tan","year":"2020","journal-title":"J. Food Sci."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"965","DOI":"10.1016\/j.talanta.2008.05.019","article-title":"Response Surface Methodology (RSM) as a Tool for Optimization in Analytical Chemistry","volume":"76","author":"Bezerra","year":"2008","journal-title":"Talanta"},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Chen, B.H., and Inbaraj, B.S. (2019). Nanoemulsion and Nanoliposome Based Strategies for Improving Anthocyanin Stability and Bioavailability. Nutrients, 11.","DOI":"10.3390\/nu11051052"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/j.foodres.2018.07.016","article-title":"Recovery of Bioactive Anthocyanin Pigments from Ficus carica L. Peel by Heat, Microwave, and Ultrasound Based Extraction Techniques","volume":"113","author":"Backes","year":"2018","journal-title":"Food Res. Int."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"590","DOI":"10.1002\/jssc.201701014","article-title":"Preparation and Characterization of Molecularly-Imprinted Polymers for Extraction of Sanshool Acid Amide Compounds Followed by Their Separation from Pepper Oil Resin Derived from Chinese Prickly Ash (Zanthoxylum bungeanum)","volume":"41","author":"Chen","year":"2017","journal-title":"J. Sep. Sci."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"696","DOI":"10.1111\/1750-3841.15037","article-title":"Extraction Optimization and Purification of Anthocyanins from Lycium ruthenicum Murr. And Evaluation of Tyrosinase Inhibitory Activity of the Anthocyanins","volume":"85","author":"Shen","year":"2020","journal-title":"J. Food Sci."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1016\/j.carbpol.2013.05.052","article-title":"Optimization of Microwave Assisted Extraction of Pectin from Orange Peel","volume":"97","author":"Prakash","year":"2013","journal-title":"Carbohydr. Polym."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"296","DOI":"10.1016\/j.foodchem.2015.07.110","article-title":"In Vitro and In Vivo Biological Activities of Anthocyanins from Nitraria tangutorun Bobr. Fruits","volume":"194","author":"Ma","year":"2016","journal-title":"Food Chem."},{"key":"ref_23","first-page":"135","article-title":"Purification Characteristics and Parameters Optimization of Anthocyanin Extracted from Blueberry","volume":"8","author":"Zheng","year":"2015","journal-title":"Int. J. Agric. Biol. Eng."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"155","DOI":"10.1016\/j.hpj.2019.01.003","article-title":"Phenolic Profile, Antioxidant Activity and Anti-proliferative Activity of Crabapple Fruits","volume":"5","author":"Han","year":"2019","journal-title":"Hortic. Plant J."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"17","DOI":"10.1016\/j.seppur.2012.11.011","article-title":"Extraction Characteristics and Optimal Parameters of Anthocyanin from Blueberry Powder under Microwave-Assisted Extraction Conditions","volume":"104","author":"Zheng","year":"2013","journal-title":"Sep. Purif. Technol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"10076","DOI":"10.1021\/jf1015347","article-title":"Pressure and Temperature Effects on Degradation Kinetics and Storage Stability of Total Anthocyanins in Blueberry Juice","volume":"58","author":"Buckow","year":"2010","journal-title":"J. Agric. Food Chem."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1016\/j.cep.2008.05.006","article-title":"Extraction, Dealcoholization and Concentration of Anthocyanin from Red Radish","volume":"48","author":"Patil","year":"2009","journal-title":"Chem. Eng. Processing Process Intensif."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"126714","DOI":"10.1016\/j.foodchem.2020.126714","article-title":"Ultrasonic-Assisted Enzymatic Extraction and Identification of Anthocyanin Components from Mulberry Wine Residues","volume":"323","author":"Zhang","year":"2020","journal-title":"Food Chem."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"81","DOI":"10.1007\/s40415-020-00590-y","article-title":"Analysis of flavonoids and anthocyanin biosynthesis-related genes expression reveals the mechanism of petal color fading of Malus hupehensis (Rosaceae)","volume":"43","author":"Han","year":"2020","journal-title":"Braz. J. Bot."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"520","DOI":"10.1016\/j.biopha.2018.05.057","article-title":"Anti-Tumor Properties of Anthocyanins from Lonicera Caerulea \u201cBeilei\u201d Fruit on Human Hepatocellular Carcinoma: In Vitro and In Vivo Study","volume":"104","author":"Zhou","year":"2018","journal-title":"Biomed. Pharmacother."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"4919","DOI":"10.2174\/1381612825666191212105145","article-title":"The Prevention and Inhibition Effect of Anthocyanins on Colorectal Cancer","volume":"25","author":"Zhao","year":"2019","journal-title":"Curr. Pharm. Des."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"7103","DOI":"10.1039\/C9FO01721F","article-title":"Isolation of Strawberry Anthocyanin-Rich Fractions and Their Mechanisms of Action against Murine Breast Cancer Cell Lines","volume":"10","author":"Mazzoni","year":"2019","journal-title":"Food Funct."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1007\/s00217-017-2956-5","article-title":"Blueberry Anthocyanin Induces Apoptosis in HepG-2 Cells and the Mechanism of the Process","volume":"244","author":"Fang","year":"2017","journal-title":"Eur. Food Res. Technol."},{"key":"ref_34","first-page":"73","article-title":"Purple Sweet Potato Anthocyanin Exerts Antitumor Effect in Bladder Cancer","volume":"40","author":"Li","year":"2018","journal-title":"Oncol. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2145","DOI":"10.1039\/C8FO00164B","article-title":"The Inhibitory Effect of Manuka Honey on Human Colon Cancer HCT-116 and LoVo Cell Growth. Part 1: The Suppression of Cell Proliferation, Promotion of Apoptosis and Arrest of the Cell Cycle","volume":"9","author":"Afrin","year":"2018","journal-title":"Food Funct."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2106","DOI":"10.1111\/j.1365-2621.2009.02044.x","article-title":"Radical Scavenging Activity and Apoptotic Effects in HT-29 Human Colon Cancer Cells of Black Sesame Seed Extract","volume":"44","author":"Kim","year":"2009","journal-title":"Int. J. Food Sci. Technol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1557","DOI":"10.1016\/j.fct.2005.05.001","article-title":"Effects of Anthocyanidin on the Inhibition of Proliferation and Induction of Apoptosis in Human Gastric Adenocarcinoma Cells","volume":"43","author":"Shih","year":"2005","journal-title":"Food Chem. Toxicol."}],"container-title":["Molecules"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/13\/4299\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T23:42:41Z","timestamp":1760139761000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1420-3049\/27\/13\/4299"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,7,4]]},"references-count":37,"journal-issue":{"issue":"13","published-online":{"date-parts":[[2022,7]]}},"alternative-id":["molecules27134299"],"URL":"https:\/\/doi.org\/10.3390\/molecules27134299","relation":{},"ISSN":["1420-3049"],"issn-type":[{"value":"1420-3049","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,4]]}}}