Article(id=1320886405295878184, tenantId=1146029695717560320, journalId=1260987677001138203, issueId=1320882506312826993, articleNumber=null, orderNo=null, doi=10.13386/j.issn1002-0306.2025060144, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1749657600000, receivedDateStr=2025-06-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1792853459904, onlineDateStr=2026-10-24, pubDate=1778774400000, pubDateStr=2026-05-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1792853459904, onlineIssueDateStr=2026-10-24, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1792853459904, creator=13701087609, updateTime=1792853459904, updator=13701087609, issue=Issue{id=1320882506312826993, tenantId=1146029695717560320, journalId=1260987677001138203, year='2026', volume='47', issue='10', pageStart='1', pageEnd='525', issueExtLink='null', onlineDate='null', pubDate='1778774400000', pubDateStr='2026-05-15', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1792852530314, creator='13701087609', updateTime=1792852530314, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext=null, issueFiles=null, downloadFileDto=null}, startPage=150, endPage=158, ext={EN=ArticleExt(id=1320886406013104169, articleId=1320886405295878184, tenantId=1146029695717560320, journalId=1260987677001138203, language=EN, title=Effects of Oil Phase on Stability and Bioaccessibility of Astaxanthin Nanoemulsions, columnId=1261336277291548795, journalTitle=Science and Technology of Food Industry, columnName=Research and Investigation, runingTitle=null, highlight=null, articleAbstract=

To enhance the bioaccessibility of astaxanthin (ASTA), this study constructed astaxanthin nanoemulsions (ASTA-NEs) using high-pressure microfluidization technology, with methyl linolenate (ML, esterified modification), flaxseed oil (FO, rich in unsaturated fatty acids), and coconut oil (CO, rich in saturated fatty acids) as the oil phases, respectively corresponding to astaxanthin-loaded methyl linolenate nanoemulsions (MLA), flaxseed oil nanoemulsions (FOA), and coconut oil nanoemulsions (COA), with astaxanthin-free methyl linolenate emulsion (ML) serving as the control. By characterizing the emulsion structure, stability, in vitro digestive behavior, and ASTA bioaccessibility, the regulatory mechanism of oil phase composition on the performance of the delivery system was revealed. Results showed that all emulsions exhibited a single narrow-peak distribution, with a polydispersity index (PDI) below 0.2 and negative Zeta potential, indicating stable and homogeneous systems. Among them, MLA had the optimal dispersion uniformity. The stability of emulsions with different oil phases was affected by pH, Na+ concentration, and temperature. Within the pH range of 5~9, MLA showed a smaller PDI value (P<0.05). For ionic stability, in the range of 0~300 mmol/L NaCl, the particle size of ML increased significantly with the increase of Na+ concentration (P<0.05), while the particle sizes of MLA, FOA, and COA remained stable. In addition, the thermal stability of different samples was evaluated within the temperature range of 30~90 ℃: At 30~60 ℃, MLA showed no significant change in particle size (P>0.05) and exhibited the best thermal stability. At 90 ℃, the particle sizes of all samples increased, but MLA still maintained a relatively small particle size (<195 nm). During in vitro digestion, the emulsion structure remained stable in the oral and gastric phases. In the intestinal phase, the particle size of COA increased significantly (P<0.05) due to the rapid hydrolysis of high saturated fatty acids. After digestion, MLA had the highest ASTA retention rate, with a bioaccessibility of 26.8%, which was significantly higher than that of FOA (24.8%) and COA (22.4%) (P<0.05). This study clarifies the regulatory mechanism of oil phase properties on the performance of astaxanthin nanoemulsions, providing a theoretical basis for the rational design and industrial application of functional lipid delivery systems.

, authors=Qianer ZHU, Xiaoqing LIU, Xiaoyu CHEN, Yue ZHANG, Chengzhen LIU*, authorsList=Qianer ZHU, Xiaoqing LIU, Xiaoyu CHEN, Yue ZHANG, Chengzhen LIU, authorCompany=null, correspAuthors=Chengzhen LIU, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright © 2026 Science and Technology of Food Industry. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, fund=null), CN=ArticleExt(id=1320886414103916604, articleId=1320886405295878184, tenantId=1146029695717560320, journalId=1260987677001138203, language=CN, title=油相对虾青素纳米乳液稳定性及生物可及性的影响, columnId=1261336277849391232, journalTitle=食品工业科技, columnName=研究与探讨, runingTitle=null, highlight=null, articleAbstract=

为提高虾青素(Astaxanthin,ASTA)的生物可及性,本研究以高压微射流技术构建由亚麻酸甲酯(ML,酯化修饰)、亚麻籽油(FO,高不饱和脂肪酸)及椰子油(CO,高饱和脂肪酸)为油相的虾青素纳米乳液(ASTA-NEs)(分别为负载虾青素的亚麻酸甲酯纳米乳液(MLA)、亚麻籽油纳米乳液(FOA)以及椰子油纳米乳液(COA)),并以不含虾青素的亚麻酸甲酯乳液(ML)作为对照。通过表征乳液结构、稳定性、消化行为及虾青素生物可及性,揭示油相组成对递送系统性能的调控机制。结果显示:所有乳液均呈单窄峰分布,多分散性指数(Polydispersity Index,PDI)低于0.2,Zeta电位为负,体系稳定均一,其中MLA分散均匀性最优。pH、Na+、温度影响不同油相乳液稳定性。在pH5~9范围内,MLA具有更小的PDI值(P<0.05);离子稳定性方面,在0~300 mmol/L NaCl范围内,ML随Na+浓度增加显著增大(P<0.05),而MLA、FOA、COA粒径保持稳定;此外,在30~90 ℃范围内评价了不同样品的热稳定性,30~60 ℃时,MLA粒径无显著变化(P>0.05),热稳定性最优,90 ℃时所有样品粒径增大,但MLA仍保持较小粒径(<195 nm)。体外消化中,口腔、胃阶段乳液结构稳定,小肠阶段COA因高饱和脂肪酸快速水解,粒径显著增大(P<0.05);MLA消化后对虾青素保留率最高,生物可及性达26.8%,显著高于FOA(24.8%)和COA(22.4%)(P<0.05)。本研究明确了油相特性对虾青素纳米乳液性能的调控机制,为功能性脂质递送系统的理性设计与工业化应用提供理论依据。

, authors=朱倩儿, 刘晓青, 陈晓宇, 张悦, 刘成珍*, authorsList=朱倩儿, 刘晓青, 陈晓宇, 张悦, 刘成珍, authorCompany=null, correspAuthors=刘成珍, authorNote=

朱倩儿(2000−),女,硕士研究生,研究方向:胶体递送,E-mail:

刘晓青(2000−),女,硕士研究生,研究方向:多功能生物材料,E-mail:

, correspAuthorsNote=
刘成珍(1991−),女,博士,副教授,研究方向:粮油精深加工、功能性成分稳态化、功能食品开发,E-mail:。
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International Journal of Biological Macromolecules, 2020, 162: 1770−1779., articleTitle=null, refAbstract=null), Reference(id=1320886841969070553, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, doi=null, pmid=null, pmcid=null, year=null, volume=null, issue=null, pageStart=null, pageEnd=null, url=null, language=null, rfNumber=40, rfOrder=39, authorNames=null, journalName=null, refType=null, unstructuredReference=OZTURK B, ARGIN S, OZILGEN M, et al. Formation and stabilization of nanoemulsion-based vitamin E delivery systems using natural surfactants: quillaja saponin and lecithin[J]. 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Comprehensive Reviews in Food Science and Food Safety, 2016, 15(2): 331−352., articleTitle=null, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1320886832129233271, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, xref=null, ext=[AuthorCompanyExt(id=1320886832137621880, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, companyId=1320886832129233271, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=College of Life Science, Qingdao University, Qingdao 266071, China), AuthorCompanyExt(id=1320886832146010489, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, companyId=1320886832129233271, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=青岛大学生命科学学院,山东青岛 266071)])], figs=[ArticleFig(id=1320886836071879071, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=EN, label=Fig.1, caption=Appearance (a), particle size distribution (b) and CLSM images (c~f) of ML, MLA, FOA and COA emulsions, figureFileSmall=6YeSkl4KhF4ZW7CMhF1zdg==, figureFileBig=TgMUhqPpQjBYk3l5lBMCAw==, tableContent=null), ArticleFig(id=1320886836138987936, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=CN, label=图1, caption=ML、MLA、FOA和COA乳液的外观(a)、粒度分布(b)和CLSM图像(c~f), figureFileSmall=6YeSkl4KhF4ZW7CMhF1zdg==, figureFileBig=TgMUhqPpQjBYk3l5lBMCAw==, tableContent=null), ArticleFig(id=1320886836218679713, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=EN, label=Fig.2, caption=Average diameter (a), PDI (b) and Zeta potential (c) of ML, MLA, FOA and COA emulsions, figureFileSmall=ptIDDJ4twvsjUxH9mgSL4w==, figureFileBig=4NaxNHTrCzlRuiAWTrMupg==, tableContent=null), ArticleFig(id=1320886836285788578, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=CN, label=图2, caption=ML、MLA、FOA和COA乳液的液滴大小(a),PDI(b)和Zeta电位(c)

注:不同大写字母表示差异显著(P<0.05),图8同。

, figureFileSmall=ptIDDJ4twvsjUxH9mgSL4w==, figureFileBig=4NaxNHTrCzlRuiAWTrMupg==, tableContent=null), ArticleFig(id=1320886836348703139, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=EN, label=Fig.3, caption=Average diameter (a), PDI (b) and Zeta potential (c) of the emulsions under different pH, figureFileSmall=cvSyYWnZgeNyG83dEQ40Ag==, figureFileBig=BbKDz5G+GWjHCPB786i5Cw==, tableContent=null), ArticleFig(id=1320886836420006308, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=CN, label=图3, caption=不同pH条件下ML、MLA、FOA和COA乳液的粒径大小(a)、PDI(b)和Zeta电位(c)

注:不同大写字母表示组间存在显著差异(P<0.05),不同小写字母表示组内存在显著差异(P<0.05),图4~图6同。

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Composition of electrolyte reserve for each part

, figureFileSmall=null, figureFileBig=null, tableContent=
组成唾液(SSF)储备液
pH7
胃液(SGF)储备液
pH3
肠液(SIF)储备液
pH7
KCl15.16.96.8
KH2PO43.70.90.8
NaHCO313.62585
NaCl13.647.238.4
MgCl2·6H2O0.150.120.33
(NH4)2CO30.060.5−
HCl1.115.68.4
), ArticleFig(id=1320886837455999408, tenantId=1146029695717560320, journalId=1260987677001138203, articleId=1320886405295878184, language=CN, label=表1, caption=

各部分的电解质储备液组成

, figureFileSmall=null, figureFileBig=null, tableContent=
组成唾液(SSF)储备液
pH7
胃液(SGF)储备液
pH3
肠液(SIF)储备液
pH7
KCl15.16.96.8
KH2PO43.70.90.8
NaHCO313.62585
NaCl13.647.238.4
MgCl2·6H2O0.150.120.33
(NH4)2CO30.060.5−
HCl1.115.68.4
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油相对虾青素纳米乳液稳定性及生物可及性的影响
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朱倩儿 , 刘晓青 , 陈晓宇 , 张悦 , 刘成珍 *
食品工业科技 | 研究与探讨 2026,47(10): 150-158
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食品工业科技 |研究与探讨 2026 , 47 (10) : 150 -158
油相对虾青素纳米乳液稳定性及生物可及性的影响
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朱倩儿 , 刘晓青 , 陈晓宇, 张悦, 刘成珍*
作者信息
  • 青岛大学生命科学学院,山东青岛 266071
通讯作者:
刘成珍(1991−),女,博士,副教授,研究方向:粮油精深加工、功能性成分稳态化、功能食品开发,E-mail:。
作者简介:

朱倩儿(2000−),女,硕士研究生,研究方向:胶体递送,E-mail:

刘晓青(2000−),女,硕士研究生,研究方向:多功能生物材料,E-mail:

Effects of Oil Phase on Stability and Bioaccessibility of Astaxanthin Nanoemulsions
Qianer ZHU , Xiaoqing LIU , Xiaoyu CHEN, Yue ZHANG, Chengzhen LIU*
Affiliations
  • College of Life Science, Qingdao University, Qingdao 266071, China
出版时间: 2026-05-15 doi: 10.13386/j.issn1002-0306.2025060144
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为提高虾青素(Astaxanthin,ASTA)的生物可及性,本研究以高压微射流技术构建由亚麻酸甲酯(ML,酯化修饰)、亚麻籽油(FO,高不饱和脂肪酸)及椰子油(CO,高饱和脂肪酸)为油相的虾青素纳米乳液(ASTA-NEs)(分别为负载虾青素的亚麻酸甲酯纳米乳液(MLA)、亚麻籽油纳米乳液(FOA)以及椰子油纳米乳液(COA)),并以不含虾青素的亚麻酸甲酯乳液(ML)作为对照。通过表征乳液结构、稳定性、消化行为及虾青素生物可及性,揭示油相组成对递送系统性能的调控机制。结果显示:所有乳液均呈单窄峰分布,多分散性指数(Polydispersity Index,PDI)低于0.2,Zeta电位为负,体系稳定均一,其中MLA分散均匀性最优。pH、Na+、温度影响不同油相乳液稳定性。在pH5~9范围内,MLA具有更小的PDI值(P<0.05);离子稳定性方面,在0~300 mmol/L NaCl范围内,ML随Na+浓度增加显著增大(P<0.05),而MLA、FOA、COA粒径保持稳定;此外,在30~90 ℃范围内评价了不同样品的热稳定性,30~60 ℃时,MLA粒径无显著变化(P>0.05),热稳定性最优,90 ℃时所有样品粒径增大,但MLA仍保持较小粒径(<195 nm)。体外消化中,口腔、胃阶段乳液结构稳定,小肠阶段COA因高饱和脂肪酸快速水解,粒径显著增大(P<0.05);MLA消化后对虾青素保留率最高,生物可及性达26.8%,显著高于FOA(24.8%)和COA(22.4%)(P<0.05)。本研究明确了油相特性对虾青素纳米乳液性能的调控机制,为功能性脂质递送系统的理性设计与工业化应用提供理论依据。

乳液  /  虾青素  /  脂肪酸组成  /  稳定性  /  体外消化  /  生物可及性

To enhance the bioaccessibility of astaxanthin (ASTA), this study constructed astaxanthin nanoemulsions (ASTA-NEs) using high-pressure microfluidization technology, with methyl linolenate (ML, esterified modification), flaxseed oil (FO, rich in unsaturated fatty acids), and coconut oil (CO, rich in saturated fatty acids) as the oil phases, respectively corresponding to astaxanthin-loaded methyl linolenate nanoemulsions (MLA), flaxseed oil nanoemulsions (FOA), and coconut oil nanoemulsions (COA), with astaxanthin-free methyl linolenate emulsion (ML) serving as the control. By characterizing the emulsion structure, stability, in vitro digestive behavior, and ASTA bioaccessibility, the regulatory mechanism of oil phase composition on the performance of the delivery system was revealed. Results showed that all emulsions exhibited a single narrow-peak distribution, with a polydispersity index (PDI) below 0.2 and negative Zeta potential, indicating stable and homogeneous systems. Among them, MLA had the optimal dispersion uniformity. The stability of emulsions with different oil phases was affected by pH, Na+ concentration, and temperature. Within the pH range of 5~9, MLA showed a smaller PDI value (P<0.05). For ionic stability, in the range of 0~300 mmol/L NaCl, the particle size of ML increased significantly with the increase of Na+ concentration (P<0.05), while the particle sizes of MLA, FOA, and COA remained stable. In addition, the thermal stability of different samples was evaluated within the temperature range of 30~90 ℃: At 30~60 ℃, MLA showed no significant change in particle size (P>0.05) and exhibited the best thermal stability. At 90 ℃, the particle sizes of all samples increased, but MLA still maintained a relatively small particle size (<195 nm). During in vitro digestion, the emulsion structure remained stable in the oral and gastric phases. In the intestinal phase, the particle size of COA increased significantly (P<0.05) due to the rapid hydrolysis of high saturated fatty acids. After digestion, MLA had the highest ASTA retention rate, with a bioaccessibility of 26.8%, which was significantly higher than that of FOA (24.8%) and COA (22.4%) (P<0.05). This study clarifies the regulatory mechanism of oil phase properties on the performance of astaxanthin nanoemulsions, providing a theoretical basis for the rational design and industrial application of functional lipid delivery systems.

emulsion  /  astaxanthin  /  fatty acid composition  /  stability  /  in vitro digestion  /  bioaccessibility
朱倩儿, 刘晓青, 陈晓宇, 张悦, 刘成珍. 油相对虾青素纳米乳液稳定性及生物可及性的影响. 食品工业科技, 2026 , 47 (10) : 150 -158 . DOI: 10.13386/j.issn1002-0306.2025060144
Qianer ZHU, Xiaoqing LIU, Xiaoyu CHEN, Yue ZHANG, Chengzhen LIU. Effects of Oil Phase on Stability and Bioaccessibility of Astaxanthin Nanoemulsions[J]. Science and Technology of Food Industry, 2026 , 47 (10) : 150 -158 . DOI: 10.13386/j.issn1002-0306.2025060144
虾青素(Astaxanthin,ASTA)作为一种天然酮式类胡萝卜素,是目前已知活性最强的天然抗氧化剂之一[1−2]。其独特的共轭双键结构与末端极性基团赋予两亲性分子构型,使其具备跨膜抗氧化、抗炎及增强免疫力等生理功能,在功能性食品、营养保健等领域具有重要应用价值[3]。然而,虾青素高度不饱和的化学结构使其在加工储存及胃肠道(GI)消化过程中易受光、热、酸碱等条件影响,严重限制了其生物利用效率与应用潜力[4−5]。因此,构建高效递送系统成为突破上述瓶颈的关键。纳米乳液(NEs)是由油相、水相及乳化剂组成的热力学稳定胶体分散体系,其分散相液滴平均粒径通常≤200 nm,作为新型脂质递送载体,通过双亲性表面活性剂形成的界面层包裹纳米尺度液滴[6],能有效改善疏水性生物活性物质的水分散性、化学稳定性及肠道吸收效率[7−8]。
近年,关于虾青素纳米乳液递送系统的研究在界面调控、载体创新及生物利用度提升方面取得显著突破。Bautista-Hernández等[9]通过大豆卵磷脂-吐温40复配乳化剂(HLB 12.1)构建高HLB值纳米乳液,首次建立表面张力梯度(Δγ)与生物可及性的量化模型,将虾青素肠道释放率提升至45.0%,但其油相局限于氢化植物油(高饱和脂肪酸);Zhang等[10]利用南极磷虾水溶性蛋白复合物(WPAK)构建高内相Pickering乳液(HIPPE 80),实现7 μm均匀粒径,经离心、盐离子(5.0% NaCl)处理及长期储存后未分层,虾青素包封率达82.6%,紫外照射6 h后保留率较游离形态提升2.3倍。Ge等[11]利用大豆分离蛋白/海藻酸钠(SPI/SA)复合物稳定纳米乳液,包封率提升至88.3%,体外消化生物可及性达47.9%。分子对接证实氢键参与复合物形成,且60 ℃储存7 d后,复合物的保留率高于90%,显著增强了乳液热稳定性及冻融分散性。然而,这些体系中油相筛选单一化,未能系统解析不同油相对递送性能的调控机制。作为乳液的核心组分,油相的理化特性(包括脂肪酸不饱和度、烃链长度、极性基团及动态黏度)及其添加比例对液滴粒径分布、界面膜厚度及系统稳定性具有决定性影响[12]。例如,直接调控液滴界面分子排列密度与微观结构影响纳米乳液功能特性[13]。其中,作为一种富含饱和脂肪链-月桂酸(C12:0,含量>45.0%)的油相,椰子油可使乳液形成刚性界面膜,赋予其优异的物理稳定性[14]。在消化过程中,脂肪酸的水解释放可加速混合胶束形成[15]。而亚麻籽油富含α-亚麻酸(C18:3n-3,含量>50.0%),其多不饱和结构增强了界面膜柔韧性和流动性,不仅能提升脂溶性成分的包载效率[16],还能通过调节脂肪酶活性控制脂质消化进程,实现包封物的缓慢释放[17]。而酯化修饰的脂肪酸(如亚麻酸甲酯)可能通过引入极性基团优化界面相互作用[18]。
基于目前缺乏不同类型油相(饱和/不饱和、酯化修饰油脂等)的系统性对比,难以明确油相特性与乳液性能及虾青素生物可及性的构效关系,且本团队前期研究发现,脂肪酸的不饱和度对小肠中胶束的形成及包封虾青素含量有显著影响,其中亚麻酸表现出最优异的相互作用力及包封效果[19]。在此基础上,本研究选取亚麻酸甲酯(ML,酯化修饰)、亚麻籽油(FO,高不饱和脂肪酸)及椰子油(CO,高饱和脂肪酸)三类油相,采用高压微射流技术构建纳米乳液递送体系。选择这三类油相可全面覆盖油相饱和度及修饰状态的差异,并以未包封虾青素的亚麻酸甲酯乳液(ML)为对照,以排除酯化油相自身对实验结果的干扰。通过系统表征乳液稳定性、体外消化行为及虾青素生物可及性,揭示油相组成对脂质基递送系统性能的调控机制。本研究成果不仅有助于深化对油相-乳液性能构效关系的理解,更为功能性脂质递送系统的理性设计与工业化应用提供关键理论依据。
虾青素、吐温80、甘油、亚麻酸甲酯、亚麻籽油、椰子油、α-淀粉酶(40000 U/g) 上海麦克林生化有限公司;猪胆盐(≥65.0%) 北京索莱宝科技有限公司;胃蛋白酶(15000 U/g)、胰酶(4000 U/g) 上海源叶生物科技有限公司;其他试剂均为分析纯。
FBZ1002-UP纯水机 青岛富勒姆集团;JES-FA200马尔文粒度仪 英国马尔文仪器有限公司;XB 220A电子天平 上海普利赛斯集团;KQ-400DE超声仪 昆山市超声仪器有限公司;MS-PA磁力搅拌器 大龙兴创实验仪器股份公司;TGL-15B高速台式离心机 上海安亭科学仪器厂;UV-6300紫外-分光光度计 上海美谱达仪器公司;LSM800高分辨率共聚焦显微镜 德国卡尔蔡司公司;T18 Digital手持均质器 德国艾卡仪器设备有限公司;NanoGenizer-II高压微射流 苏州微流纳米生物技术有限公司。
依据Xu等[20]的方法并进行部分修改制备乳液。
油相制备:取0.006 g虾青素分别溶于4 g亚麻酸甲酯、亚麻籽油、椰子油中(w/w),使用磁力搅拌器(40 ℃,30 min)使其完全溶解,制得油相。
水相制备:取1.2 g甘油(w/w)及1.2 g吐温80(w/w)溶于33.6 g去离子水,混合均匀即得到水相。
乳液制备:将上述油相(10%(w/w))缓慢加入到水相(90%(w/w))中,边加边使用手持均质机制备粗乳(14000 r/min,10 min)。将制备得到的粗乳在100 bar下通过高压微射流5次,即制备得到负载虾青素的亚麻酸甲酯纳米乳液(MLA)、负载虾青素的亚麻籽油纳米乳液(FOA)和负载虾青素的椰子油纳米乳液(COA),并制备不添加虾青素的亚麻酸甲酯乳液(ML)作为对照组。为避免虾青素氧化,全程均避光操作。
使用高分辨率共聚焦显微镜(Confocal laser scanning microscope,CLSM)观察乳液的表观形态。根据Xia等[21]的方法稍作修改。将1 mg尼罗红染料溶于1 mL乙醇溶液中,取乳液样品稀释10倍后,使用尼罗红溶液对乳液的油相染色。取20 μL尼罗红溶液加入1 mL稀释后的乳液,混合均匀,负染5 min,然后将样品置于载玻片上,盖上盖玻片,在激发波长为488 nm处进行显微观察。
根据Inthamat等[22]的方法,取0.1 mL乳液与9.9 mL去离子水混合,稀释后将分散的乳液样品放入马尔文粒度仪中。设置参数:温度25 ℃,分散相为水(折射率1.33)。测定乳液样品的粒径、Zeta电位和多分散指数。
配制0.1 mol/L的HCl和NaOH,将各组样品的pH分别调整至3、5、7、9、11,室温孵育30 min,测定不同pH下样品的粒径、Zeta电位和多分散指数[23],方法同1.2.3。
用NaCl溶液调节样品的盐离子浓度梯度为0、50、100、200、300 mmol/L的样品,室温孵育30 min后,测定不同盐离子浓度条件下样品的粒径、Zeta电位。用水稀释100倍测定其粒径、Zeta电位和多分散指数[24],方法同1.2.3。
根据Xing等[25]和Zhong等[26]的方法,稍作修改,将5 mL ML、MLA、FOA、COA放入琥珀色玻璃瓶中,分别在30、60、90 ℃三种条件下水浴孵育30 min。待冷却至室温后,测定不同温度下样品的粒径、Zeta电位和多分散指数,方法同1.2.3。
采用INFOGEST体外消化模型,稍作修改[27]。模型由口、胃、肠三部分组成,各部分的电解质储备液组成如表1所示。
口腔阶段:取5 mL待消化样液,向其依次加入4 mL SSF储备液、0.075 g的α-淀粉酶、20 µL 0.3 mol/L CaCl2溶液和975 µL的水,混合后在摇床上(37 ℃,100 r/min)振荡2 min。
胃阶段:取10 mL口腔阶段的样品,依次加入8 mL SGF储备液、5.3 g胃蛋白酶和5 µL的氯化钙溶液,并加水至20 mL,将pH调整至3.0,混合后在摇床上(37 ℃,100 r/min)振荡120 min。
小肠阶段:向20 mL胃阶段的样品中加入13 mL SIF储备液、1 g胰酶和40 µL 0.3 mol/L CaCl2溶液,以及溶于3 mL SIF储备液的胆盐(0.001 mol/L)。并加水至40 mL,将pH调整至7.0,混合后在摇床上(37 ℃,100 r/min)振荡120 min。用0.45 μm滤膜过滤去除杂质,即得到各样品组的混合胶束相。
使用CLSM观察乳液的表观形态,方法同1.2.2。
使用马尔文粒度仪测定乳液的粒径、电位和多分散指数,方法同1.2.3。
分别取1 mL的ML、MLA、FOA、COA及各组样品消化后的混合胶束相,将其与4 mL溶剂混合(二氯甲烷:甲醇=2:1),离心(25 ℃,5000 r/min,15 min),取有机相,用紫外-分光光度计在488 nm处测定其吸光度[21],并用同样的有机溶剂配制不同浓度的虾青素,制备标曲,其线性回归方程为:y=0.20138x−0.31182(R2=0.99984)。依据以下公式对乳液的生物可及性进行计算[24]。
$\rm 生物可及性(\text{%})=\frac{{C}_{\text{micelle}}}{C_{\text{0}}}\times 100 $
式中:Cmicelle表示上清液中虾青素含量,mg/mL;C0表示乳剂中总虾青素含量,mg/mL。
所有的实验均进行3次平行实验。结果表示为平均值±标准差。用SPSS软件(SPSS 20,Chicago,IL)对实验数据进行统计分析,进行单因素方差One-way ANOVA分析,并采用Tukey’s法进行两两比较分析,P<0.05被认为具有统计学意义,绘图采用Origin 2021软件。
通过高压微射流制备得到的乳液外观如图1a所示,ML呈现乳白色,而MLA、FOA和COA因负载虾青素,外观颜色与ML呈现明显差异。另外MLA、FOA、COA外观存在差异,可能是因为油相化学结构与特性影响了虾青素的分散状态与界面膜性质。如图1b所示,四种乳液样品均呈单窄峰分布,说明其分布均匀[28]。采用CLSM拍摄四种乳液体系如图1c~图1f所示,进一步分析乳液的微观形貌。观察发现,所有样品中液滴边界均呈现出清晰的尼罗红荧光标记轮廓,表明乳化剂分子在油-水界面形成了致密的吸附层,属于O/W型乳液[29],这种致密的界面膜是维持乳液物理稳定性的关键屏障[30]。结合粒径分布结果(图1b),CLSM图像直观地证实了各体系液滴尺寸均一性好、空间分布均匀。这种微观尺度上的均匀性对于维持乳液的长期物理稳定性具有重要意义[31]。值得注意的是,对比不同样品,ML及MLA乳液的液滴在视野中的分散性更好,分布更均匀。这种差异可能与油相本身的物理化学性质(如黏度、极性)及其与乳化剂Tween 80的相互作用有关[32]。
如图2a所示,ML的平均粒径为173.3 nm,而负载虾青素的乳液体系中MLA、FOA和COA的平均粒径分别为173.0、212.6和202.0 nm。负载虾青素后,亚麻酸甲酯乳液的液滴尺寸略有减小,这种粒径的减小可能与类胡萝卜素分子嵌入所引起的结构压缩有关[32–33],或者与更强相互作用力有关[34]。与ML和MLA相比,FOA和COA的平均粒径更大(P<0.05),但整体仍保持在纳米尺度范围内。如图2b所示,所有样品的PDI均低于0.2,表明液滴粒径分布狭窄,体系具有高度均一性,有利于乳液的长期稳定性[35],其中MLA具有最好的液滴分散均匀性(P<0.05)。就MLA来说,虾青素的负载几乎没有改变乳液的颗粒表面电荷。亚麻酸甲酯作为油相制备得到的乳液表现出良好的分散性和较小的粒径,得到了Jaromin等[34]研究的支持,其使用从亚麻籽油中获得的多不饱和脂肪酸n-3和n-6的乙酯作为油相制备O/W型纳米乳液,其液滴平均粒径介于117.7~133.3 nm之间,多分散指数(PDI)均低于0.2,展现出优异的分散均一性,与本文乳液的结构优势一致。除粒径和PDI参数外,Zeta电位对纳米乳液的稳定性及功能效率具有重要影响[36]。研究表明,负Zeta值可通过颗粒间静电斥力维持分散体系稳定,且其绝对值通常与稳定性正相关,绝对值越大,静电斥力越强,体系越稳定[36–37]。如图2c所示,四种乳液的Zeta均表现出负值,不存在显著差异,表明四种乳液在储存过程中均具有良好的稳定性[38]。
乳液的pH稳定性通过粒径、PDI和Zeta电位进行评估。如图3所示,除pH=11时,四种乳液的平均粒径在不同pH条件下基本未见显著性差异,表明在宽pH范围内具有良好的物理稳定性。PDI虽略有波动,但基本维持在0.2左右,进一步证实了其均一性不受pH变化的明显影响。这种稳定性主要归因于所使用的非离子型乳化剂吐温80,其乳化性能受pH影响较小,能够在较宽的pH范围内有效维持乳液体系[39],其中,MLA在pH=9时具有最小的PDI。Zeta电位的变化则反映了乳液颗粒表面电荷随pH环境的调整,且不同油相制备的乳液电位表现出显著差异。在pH=3时,Zeta电位接近中性,表明此时油滴表面可解离基团的电离程度很低。随着pH的升高,溶液中OH-浓度增加,颗粒表面官能团解离加剧,负电荷大量增加,导致电位绝对值均显著增大(P<0.05)[39]。值得注意的是,油相本身的化学组成是导致电位差异的关键因素:富含不饱和脂肪酸的亚麻籽油虾青素纳米乳液FOA中,不饱和脂肪酸链呈弯曲构象,这一结构特点有利于羧基的暴露与解离,因此该纳米乳液的 Zeta 电位绝对值最大;与之相比,富含饱和脂肪酸的椰子油虾青素纳米乳液COA,其直链脂肪酸的紧密排列可能限制了羧基的解离程度,导致电位绝对值较小;而经过酯化修饰的亚麻酸甲酯虾青素纳米乳液MLA,由于失去了可解离的羧基(-COOH),其电位主要来源于乳化剂贡献,因此其电位绝对值最小[40−42]。相比于ML,MLA的电位绝对值虽然有所降低,但虾青素的负载有可能引入了疏水相互作用等除静电引力外的其他相互作用[42–43],使其具有更小的PDI,因此表现出更均一的分布。油相特性引起的Zeta电位差异,直接影响着乳液颗粒间的静电排斥力,是决定其胶体稳定性的重要因素,也为深入理解油相性质与乳液性能之间的构效关系提供了关键依据。
四种乳液在不同Na+浓度下的平均粒径、PDI和电位如图4所示。ML的粒径随着Na+浓度的增加而显著增加(P<0.05),表明ML乳液受钠离子影响最大,这主要归因于ML缺乏天然油脂中的内源性抗氧化保护,导致其界面膜在高离子强度环境下更易受氧化劣化和静电屏蔽作用的影响[44]。相比之下,负载有虾青素的乳液均具有对高浓度钠离子的抵抗力。这表明虾青素的负载一定程度上改变了乳液界面性质,影响了其对钠离子浓度变化的响应。观察PDI变化,四种乳液的PDI在不同NaCl浓度下虽有波动,但总体维持在较低水平,表明钠离子浓度变化未显著破坏乳液粒径分布的均匀性,体系仍保持相对稳定。随着NaCl浓度的增加,四种乳液的Zeta电位绝对值总体上都显著降低(P<0.05)。钠离子浓度的增加可能压缩了乳液液滴的双电层,降低了液滴之间的静电排斥力[22]。MLA和ML的Zeta电位在不同NaCl浓度下变化相似,但在高浓度NaCl时,MLA的Zeta电位绝对值显著高于ML(P<0.05)。这表明虾青素的负载可能对乳液的稳定性产生了一定的影响。虾青素作为抗氧化剂,可能在一定程度上保护了乳液中的脂类物质不被氧化,从而在高浓度钠离子存在下维持了相对较高的稳定性[22]。
四种乳液的热稳定性结果如图5所示,在90 ℃处理下,四种乳液的平均粒径存在显著差异(P<0.05),其大小顺序为:FOA>COA>MLA>ML。其中,ML和MLA乳液展现出更小的平均粒径和较低的PDI,表明其在高温条件下具有更好的物理稳定性。考察温度梯度变化的影响发现:当温度从30 ℃升至60 ℃时,FOA乳液的粒径由215.4±1.0 nm显著减小至211.8±2.0 nm(P<0.05)。COA乳液的粒径由199.8±3.0 nm变化至202.1±5.0 nm,MLA乳液的粒径由192.5±0.4 nm变化至192.2±2.3 nm,二者均未发生显著变化。相比之下,MLA在此温度区间(30~60 ℃)表现出最优异的热稳定性。进一步分析Zeta电位随温度的变化(图5c),四种乳液在不同温度下的电位值均存在显著差异(P<0.05),并呈现一定的规律性,与30 ℃时的电位值相比,MLA、FOA和COA乳液在60 ℃时的Zeta电位绝对值均呈现小幅上升趋势;而当温度进一步升高至90 ℃时,这三种乳液的Zeta电位绝对值则出现小幅下降。ML乳液的电位变化模式略有不同。这些电位变化反映了乳液界面性质在升温过程中的动态调整。综合粒径与电位结果,四种乳液中,ML和MLA,尤其是MLA,在经受高温考验时表现出相对更佳的稳定性。吐温80作为非离子乳化剂,其稳定的界面膜特性可能对维持乳液在宽温度范围内的稳定性起到了关键作用[45]。
四种乳液在消化各阶段(口、胃、小肠)以及最终形成的混合胶束的粒径、PDI和Zeta电位如图6所示。口腔阶段,四种乳液粒径和PDI均较小且差异不显著,表明口腔环境对乳液结构影响微弱,乳液保持相对稳定,CLSM(图7)结果与此一致。胃阶段,各乳液粒径和PDI变化幅度小,说明胃蛋白酶等条件未对乳液粒径产生明显影响,结构维持稳定。进入小肠阶段,受胰酶等作用,乳液结构开始改变,粒径和PDI出现不同程度变化。与此相对应,Zeta电位结果显示,胃环境使所有样品负电荷密度减小,电位绝对值显著降低(P<0.05),而小肠与胶束环境则促使负电荷密度增大,电位绝对值显著升高(P<0.05);其中COA在小肠和胶束环境中表现出最大的Zeta电位绝对值(30~32 mV),显著高于另外三组(P<0.05),揭示其在肠道及后续吸收阶段可能具有更优异的胶体稳定性。CLSM(图7)显示小肠阶段出现明显的聚集,其中COA的粒径和 PDI 增幅显著大于另外三组(P<0.05)。这是由于椰子油富含中链高饱和脂肪酸,快速水解生成中链脂肪酸,使乳液界面膜结构发生剧烈改变,液滴间相互作用失衡,部分液滴融合变大,部分破碎变小,导致粒径显著变化且分布不均(PDI波动大)[15−16]。而其他乳液的油相(如亚麻酸甲酯、亚麻籽油),其脂肪酸结构(酯化修饰或高不饱和结构)使水解速率与产物对界面膜的影响较小[17],界面膜相对稳定,故另外三组的粒径和 PDI 变化明显小于 COA 组。
为了研究不同油相乳液对虾青素生物可及性的影响,首先建立了虾青素浓度与吸光度之间的标准曲线,计算后发现,不同油相乳液中虾青素的生物可及性存在显著差异(P<0.05),MLA的生物可及性最高(26.8%),依次是FOA(24.8%)和COA(22.4%),如图8所示。MLA、FOA、COA的虾青素生物可及性差异与其消化稳定性相关。MLA在消化中粒径、PDI波动小,结构稳定,小肠阶段微观结构完整,利于虾青素释放吸收,故生物可及性最高。COA含高饱和脂肪酸的椰子油,小肠消化时界面膜水解相对剧烈,结构破坏严重,稳定性差,虾青素生物可及性最低。综上,油相组成显著影响虾青素的生物可及性,其中MLA可有效提高虾青素的释放与吸收,而COA则表现出最低的生物可及性。
本研究通过高压微射流技术成功制备了负载虾青素的纳米乳液(MLA、FOA、COA)及空白乳液(ML),系统分析了其理化性质,环境稳定性(pH、离子强度、温度)及生物可及性。结果表明,虾青素的负载显著影响乳液外观颜色,但对粒径分布及Zeta电位影响较小,所有乳液均呈现纳米级液滴(D50≈173~213 nm)及高度均一性(PDI<0.2)。在pH稳定性测试中,非离子型乳化剂吐温80赋予乳液宽pH适应性,其粒径及PDI在pH3~9范围内保持稳定,而Zeta电位随pH升高负电荷增强,印证了界面官能团的解离调控机制。离子强度稳定性测试显示,虾青素的引入提升了乳液对高离子强度的耐受性,MLA的Zeta电位在高盐条件下仍维持较高绝对值(>5 mV),表明其界面膜稳定性最优。热稳定性实验中,负载虾青素的乳液在90 ℃下粒径变化幅度显著小于ML(P<0.05),其中MLA在30~60 ℃粒径无显著变化,90 ℃时仍维持<195 nm的较小粒径;而ML因缺乏虾青素与界面分子的疏水协同作用,高温下粒径收缩更明显。体外消化研究进一步证实,油相化学结构(链长、不饱和度、修饰状态)显著调控乳液消化行为与生物可及性:亚麻酸甲酯虾青素纳米乳液(MLA)在消化过程中粒径和PDI变化相对较小,结构较为稳定,生物可及性达26.8%;亚麻籽油虾青素纳米乳液(FOA)次之(24.8%);而椰子油虾青素纳米乳液(COA)因小肠阶段脂肪酸快速水解导致结构破坏,生物可及性最低(22.4%)。综上,本研究阐明了油相链长、不饱和度和虾青素负载与乳液稳定性间的构效关系,为开发高稳定性、高生物可及性的脂溶性活性成分递送系统提供了理论依据与技术参考。

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2026年第47卷第10期
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doi: 10.13386/j.issn1002-0306.2025060144
  • 接收时间:2025-06-12
  • 首发时间:2026-10-24
  • 出版时间:2026-05-15
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  • 收稿日期:2025-06-12
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    青岛大学生命科学学院,山东青岛 266071

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刘成珍(1991−),女,博士,副教授,研究方向:粮油精深加工、功能性成分稳态化、功能食品开发,E-mail:。
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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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