{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,31]],"date-time":"2026-10-31T01:29:10Z","timestamp":1793410150639,"version":"4.5.0"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2025,5,7]],"date-time":"2025-05-07T00:00:00Z","timestamp":1746576000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"the Shanxi Provincial Guiding Special Program for Commercialization of Scientific and Technological Achievements","award":["202304021301031"],"award-info":[{"award-number":["202304021301031"]}]},{"name":"Dalian Key Research and Development Program Projects","award":["2024YF12GX011"],"award-info":[{"award-number":["2024YF12GX011"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:sec id=\"sec001\">\n<jats:title>Objective<\/jats:title>\n<jats:p>Hempseed threshing residues are rich in phytochemicals such as polyphenols and flavonoids. Phenolic and flavonoid compounds have been associated with antioxidant, antibacterial and anticancer activities. The re-use of the hempseed threshing residues as value-added materials is, not only cost-saving, but also environmentally beneficial. It is therefore important to develop an effective method for extraction of phenolic compounds and flavonoids from hempseed threshing residues.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec002\">\n<jats:title>Methods<\/jats:title>\n<jats:p>In this investigation, the extraction of phenolic constituents and flavonoids from hempseed threshing residues using heat reflux extraction (HRE) were optimized through response surface methodology (RSM). Four HRE parameters to enhance the yield of crude extracts (CE), total phenolic content (TPC), and total flavonoids content (TFC) were evaluated. Additionally, the study evaluated the chemical compounds, antioxidant characteristics of the extracts, and the immune activity of the extracts was assessed by quantifying the levels of inflammatory cytokines, specifically IL-6, IL-10, and TNF-\u03b1.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec003\">\n<jats:title>Results<\/jats:title>\n<jats:p>The best extraction parameters were determined as: for the extraction time of 69.71 min, a liquid-solid proportion of 5.12:1, a particle size of 1150 \u00b5m, and an ethanol concentration of 69.60%. Under these optimized conditions, the yields for CE, TPC, and TFC were 4.74%, 27.54%, and 16.02% respectively. The data conformed well to multiple regression models, showing that these extraction parameters markedly influence the yields of CE, TPC, and TFC. Most of the compounds found may belong to the class of polyphenol and flavonoids. Cellular assays indicated that extracts from hempseed threshing residue notably reduced pro-inflammatory factors (TNF-\u03b1, IL-6) and increased anti-inflammatory factors (IL-10) in RAW 264.7 cells.<\/jats:p>\n<\/jats:sec>\n<jats:sec id=\"sec004\">\n<jats:title>Conclusion<\/jats:title>\n<jats:p>This research lays a theoretical foundation for extracting polyphenols and flavonoids from hempseed threshing residue and for the comprehensive assessment of antioxidant and immune-enhancing products. However, the antioxidant and immune activity of hempseed threshing residues extracts under physiological conditions in vivo, and the relevant mechanism should be further studied.<\/jats:p>\n<\/jats:sec>","DOI":"10.1371\/journal.pone.0322134","type":"journal-article","created":{"date-parts":[[2025,5,7]],"date-time":"2025-05-07T17:49:31Z","timestamp":1746640171000},"page":"e0322134","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":4,"title":["Effects of heat reflux extraction on the content, antioxidant, and immune activity of polyphenols and flavonoids from hempseed threshing 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L.) seed oil extraction with pressurized n-propane and supercritical carbon dioxide","volume":"143","author":"DR Grij\u00f3","year":"2019","journal-title":"J Supercrit Fluids"},{"issue":"18","key":"pone.0322134.ref005","doi-asserted-by":"crossref","first-page":"3505","DOI":"10.3390\/foods12183505","article-title":"The role of hemp (Cannabis sativa L.) as a functional food in vegetarian nutrition","volume":"12","author":"DR Grij\u00f3","year":"2023","journal-title":"Foods"},{"key":"pone.0322134.ref006","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1016\/j.foodchem.2017.09.080","article-title":"Biorefining of industrial hemp (Cannabis sativa L.) threshing residues into cannabinoid and antioxidant fractions by supercritical carbon dioxide, pressurized liquid and enzyme-assisted extractions","volume":"267","author":"V Kitryt\u0117","year":"2018","journal-title":"Food 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Arora","year":"2022"},{"key":"pone.0322134.ref010","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1002\/9783527844494.ch12","article-title":"Comparative studies on thermal,microwave\u2010assisted, and ultrasound\u2010promoted preparations.","volume-title":"Green chemical synthesis with microwaves and ultrasound","author":"TP Adhi","year":"2024"},{"issue":"9","key":"pone.0322134.ref011","doi-asserted-by":"crossref","first-page":"3077","DOI":"10.1007\/s11696-020-01140-2","article-title":"Comparison of different extraction techniques and optimization of the microwave-assisted extraction of saponins from Aralia elata (Miq.) Seem fruits and rachises","volume":"74","author":"Y Tian","year":"2020","journal-title":"Chem Pap"},{"issue":"4","key":"pone.0322134.ref012","doi-asserted-by":"crossref","first-page":"4140","DOI":"10.1007\/s11694-023-01951-3","article-title":"Heat-reflux processing of black peppercorn into bioactive antioxidant oleoresins: a three-functioned Taguchi-based grey relational grading","volume":"17","author":"OA Olalere","year":"2023","journal-title":"Food Measure"},{"key":"pone.0322134.ref013","doi-asserted-by":"crossref","first-page":"101479","DOI":"10.1016\/j.scp.2024.101479","article-title":"Green and efficient approach to extract bioactive flavonoids with antioxidant, antibacterial, antiglycation, and enzyme inhibitory activities from navel orange peel","volume":"38","author":"C Lai","year":"2024","journal-title":"Sustain Chem Pharm"},{"issue":"10","key":"pone.0322134.ref014","doi-asserted-by":"crossref","first-page":"3010","DOI":"10.1021\/jf0115589","article-title":"Thermal processing enhances the nutritional value of tomatoes by increasing total antioxidant activity","volume":"50","author":"V Dewanto","year":"2002","journal-title":"J Agric Food Chem"},{"issue":"4","key":"pone.0322134.ref015","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1016\/S0308-8146(98)00102-2","article-title":"The determination of flavonoid contents in mulberry and their scavenging effects on superoxide radicals","volume":"64","author":"Z Jia","year":"1999","journal-title":"Food Chem"},{"issue":"9","key":"pone.0322134.ref016","doi-asserted-by":"crossref","first-page":"4156","DOI":"10.1021\/jf000227d","article-title":"Radical-scavenging activities of citrus essential oils and their components: detection using 1,1-diphenyl-2-picrylhydrazyl","volume":"48","author":"HS Choi","year":"2000","journal-title":"J Agric Food Chem"},{"issue":"1\u20133","key":"pone.0322134.ref017","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1016\/j.bbagen.2004.08.006","article-title":"Reevaluating the role of 1,10-phenanthroline in oxidative reactions involving ferrous ions and DNA damage","volume":"1675","author":"IGJ de Avellar","year":"2004","journal-title":"Biochim Biophys Acta"},{"issue":"1","key":"pone.0322134.ref018","doi-asserted-by":"crossref","first-page":"45","DOI":"10.1016\/j.fct.2003.08.004","article-title":"Antioxidant capacity of reaction products limits the applicability of the Trolox Equivalent Antioxidant Capacity (TEAC) assay","volume":"42","author":"MJTJ Arts","year":"2004","journal-title":"Food Chem Toxicol"},{"issue":"1","key":"pone.0322134.ref019","doi-asserted-by":"crossref","first-page":"70","DOI":"10.1006\/abio.1996.0292","article-title":"The ferric reducing ability of plasma (FRAP) as a measure of \u201cantioxidant power\u201d: the FRAP assay","volume":"239","author":"IF Benzie","year":"1996","journal-title":"Anal Biochem"},{"key":"pone.0322134.ref020","doi-asserted-by":"crossref","first-page":"380","DOI":"10.1016\/j.ijbiomac.2021.06.026","article-title":"Structural characterization, anticancer, hypoglycemia and immune activities of polysaccharides from Russula virescens","volume":"184","author":"Y-M Li","year":"2021","journal-title":"Int J Biol Macromol"},{"issue":"2","key":"pone.0322134.ref021","doi-asserted-by":"crossref","first-page":"372","DOI":"10.1080\/10942912.2016.1163266","article-title":"Aqueous extraction kinetics of phenolic compounds from jamun (Syzygium cuminiL.) seeds","volume":"20","author":"U Balyan","year":"2016","journal-title":"Int J Food Prop"},{"issue":"1","key":"pone.0322134.ref022","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/j.foodchem.2004.07.015","article-title":"Optimization of continuous phenol extraction from byproducts","volume":"92","author":"M Pinelo","year":"2005","journal-title":"Food Chem"},{"key":"pone.0322134.ref023","first-page":"113","article-title":"Optimization of ultrasound-assisted extraction of Helicteres hirsuta Lour. for enhanced total phenolic compound and antioxidant yield","volume":"7","author":"HNT Pham","year":"2017","journal-title":"J Appl Res Med Aromatic Plants"},{"issue":"2","key":"pone.0322134.ref024","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.1016\/j.foodchem.2013.04.003","article-title":"Optimisation of ultrasonic-assisted extraction of antioxidant compounds from Artemisia absinthium using response surface methodology","volume":"141","author":"S Sahin","year":"2013","journal-title":"Food Chem"},{"key":"pone.0322134.ref025","doi-asserted-by":"crossref","first-page":"382","DOI":"10.1016\/j.indcrop.2018.07.070","article-title":"Optimization of ultrasound-assisted extraction of phenolic compounds: oleuropein, phenolic acids, phenolic alcohols and flavonoids from olive leaves and evaluation of its antioxidant activities","volume":"124","author":"M Irakli","year":"2018","journal-title":"Ind Crops Prod"},{"key":"pone.0322134.ref026","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/j.fbp.2013.10.004","article-title":"Reduced pressure extraction of oleuropein from olive leaves (Olea europaea L.) with ultrasound assistance","volume":"93","author":"P Xie","year":"2015","journal-title":"Food Bioprod Proces"},{"issue":"5","key":"pone.0322134.ref027","doi-asserted-by":"crossref","first-page":"1523","DOI":"10.1016\/j.jpba.2006.04.002","article-title":"Phenolics in cereals, fruits and vegetables: Occurrence, extraction and analysis","volume":"41","author":"M Naczk","year":"2006","journal-title":"J Pharmaceut Biomed"},{"issue":"11","key":"pone.0322134.ref028","doi-asserted-by":"crossref","first-page":"3064","DOI":"10.1007\/s11947-014-1313-y","article-title":"The use of microwave and pulsed electric field as a pretreatment step in ultrasonic extraction of polyphenols from defatted hemp seed cake (Cannabis sativa) using response surface methodology","volume":"7","author":"S-S Teh","year":"2014","journal-title":"Food Bioprocess Technol"},{"issue":"10","key":"pone.0322134.ref029","doi-asserted-by":"crossref","first-page":"1319","DOI":"10.1002\/aocs.12108","article-title":"Effect of solvent, preheating temperature, and time on the ultrasonic extraction of phenolic compounds from cold\u2010pressed hempseed cake","volume":"95","author":"J Liang","year":"2018","journal-title":"J Americ Oil Chem Soc"},{"issue":"5","key":"pone.0322134.ref030","doi-asserted-by":"crossref","first-page":"4057","DOI":"10.1039\/D0FO00066C","article-title":"Characterization of bioactive compounds in defatted hempseed (Cannabis sativa L.) by UHPLC-HRMS\/MS and anti-inflammatory activity in primary human monocytes","volume":"11","author":"J Rea Martinez","year":"2020","journal-title":"Food Funct"},{"issue":"4","key":"pone.0322134.ref031","doi-asserted-by":"crossref","first-page":"3409","DOI":"10.1007\/s10311-021-01217-8","article-title":"Techniques and modeling of polyphenol extraction from food: a review","volume":"19","author":"A Sridhar","year":"2021","journal-title":"Environ Chem Lett"},{"issue":"4","key":"pone.0322134.ref032","doi-asserted-by":"crossref","first-page":"2122","DOI":"10.1016\/j.foodchem.2012.11.099","article-title":"Application of response surface methodology to optimise extraction of flavonoids from fructus sophorae","volume":"138","author":"Q Xu","year":"2013","journal-title":"Food Chem"},{"key":"pone.0322134.ref033","first-page":"101084","article-title":"Chemical and functional characterization of the main bioactive molecules contained in hulled Cannabis sativa L. seeds for use as functional ingredients","volume":"16","author":"S Frazzini","year":"2024","journal-title":"J Agric Food Res"},{"issue":"7","key":"pone.0322134.ref034","doi-asserted-by":"crossref","first-page":"3159","DOI":"10.1111\/1750-3841.15813","article-title":"Effect of extrusion technology on hempseed (Cannabis sativa L.) oil cake: Polyphenol profile and biological activities","volume":"86","author":"W Leonard","year":"2021","journal-title":"J Food Sci"},{"key":"pone.0322134.ref035","doi-asserted-by":"crossref","first-page":"543","DOI":"10.1016\/j.foodchem.2014.09.110","article-title":"Optimisation of ultrasonic-assisted extraction of phenolic compounds, antioxidants, and anthocyanins from sugar beet molasses","volume":"172","author":"M Chen","year":"2015","journal-title":"Food Chem"},{"key":"pone.0322134.ref036","doi-asserted-by":"crossref","first-page":"154","DOI":"10.1016\/j.indcrop.2019.04.001","article-title":"Phenolic composition and antioxidant, antimicrobial and cytotoxic properties of hop (Humulus lupulus L.) seeds","volume":"134","author":"JI Alonso-Esteban","year":"2019","journal-title":"Ind Crops Prod"},{"key":"pone.0322134.ref037","doi-asserted-by":"crossref","first-page":"155066","DOI":"10.1016\/j.cyto.2020.155066","article-title":"Importance of TNF-alpha and its alterations in the development of cancers","volume":"130","author":"P Mahdavi Sharif","year":"2020","journal-title":"Cytokine"},{"issue":"10","key":"pone.0322134.ref038","doi-asserted-by":"crossref","first-page":"986","DOI":"10.3390\/antiox9100986","article-title":"Antioxidant and anti-inflammatory activities in relation to the flavonoids composition of pepper (Capsicum annuum L.)","volume":"9","author":"S-Y Cho","year":"2020","journal-title":"Antioxidants (Basel)"},{"issue":"1","key":"pone.0322134.ref039","doi-asserted-by":"crossref","first-page":"511","DOI":"10.1080\/09540105.2022.2098935","article-title":"Antioxidant and anti-inflammatory activities of Gynura procumbens flowers extract through suppressing LPS-induced MAPK\/NF-\u03baB signalling 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