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Article(id=1153986653052986044, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986642063905290, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241111006, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1731254400000, receivedDateStr=2024-11-11, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1753061458123, onlineDateStr=2025-07-21, pubDate=1739548800000, pubDateStr=2025-02-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753061458123, onlineIssueDateStr=2025-07-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753061458123, creator=13701087609, updateTime=1753061458123, updator=13701087609, issue=Issue{id=1153986642063905290, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='3', pageStart='1', pageEnd='316', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1753061455502, creator=13701087609, updateTime=1760070725729, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1183385652272968023, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986642063905290, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1183385652272968024, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986642063905290, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=130, endPage=138, ext={EN=ArticleExt(id=1153986654126727889, articleId=1153986653052986044, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Preparation and characterization of gel from Chuzhou chrysanthemum polysaccharide CCPS-3-1, columnId=1151895322591638525, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Functional Foods and Functional Components, runingTitle=null, highlight=null, articleAbstract=

Objective To prepare Chuzhou chrysanthemum polysaccharide CCPS-3-1 gel and investigate its relevant properties. Methods Acidic polysaccharide CCPS-3-1 was isolated and purified from Chuzhou chrysanthemum and mixed with CaCl2 solution to form a gel via ionic crosslinking. The gel's characteristics were comprehensively analyzed using scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, and differential scanning calorimetry. Additionally, treatment with urea and ethylenediaminetetraacetic acid was employed to further elucidate its gel strength and gelling mechanism. Results CCPS-3-1 solution was relatively stable within a pH range of 6.0-8.0. Upon induction by CaCl2, the CCPS-3-1 gel exhibited good transparency, moderate hardness, and high stability. The gel network had a dense, smooth, and uniform structure. Moreover, the thermal stability and crystalline structure were enhanced. Rheological experiments revealed that as the CaCl2 concentration increased, the apparent viscosity and storage modulus of the CCPS-3-1 gel also increased. In addition, the loss tangent (Tanδ) decreased with increasing CaCl2 concentration, indicating the formation of weak gels at all tested concentrations. Furthermore, electrostatic interactions played a crucial role in the formation of the gel network structure. Conclusion The Chuzhou chrysanthemum polysaccharide CCPS-3-1, isolated and purified in this study, forms a gel upon CaCl2 induction. Its gelling mechanism likely involves the synergistic effects of intermolecular hydrogen bonding and Ca2+ crosslinking. These findings provide a theoretical foundation for the application of CCPS-3-1 gel in food, pharmaceutical, and other fields.

, correspAuthors=Chun-Xu CHEN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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, authorCompany=null, fund=null, authors=null, authorsList=Wen-Ting JI, Tian-Xiang HAN, You-Mei XU, Ming LIU, Chun-Xu CHEN), CN=ArticleExt(id=1153986701065183525, articleId=1153986653052986044, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=滁菊多糖CCPS-3-1凝胶制备及其特性研究, columnId=1151895323909124661, journalTitle=食品安全质量检测学报, columnName=本期专题：功能性食品与功能性成分, runingTitle=null, highlight=null, articleAbstract=

目的 制备滁菊多糖CCPS-3-1凝胶, 并研究其相关特性。方法 以滁菊为原料, 分离纯化得到酸性多糖CCPS-3-1, 与CaCl2溶液混合, 通过离子交联作用形成凝胶。运用扫描电子显微镜、傅里叶红外光谱、X-射线衍射、差示扫描量热法等测定方法对其形成的凝胶特性进行全面分析, 并用尿素和乙二胺四乙酸处理进一步阐明其胶凝力和胶凝机制。结果 在pH 6.0~8.0范围内, CCPS-3-1溶液相对稳定。经CaCl2诱导后, CCPS-3-1凝胶呈现出良好的透明度, 有一定的硬度和良好的稳定性, 凝胶网络结构致密, 表面光滑且均匀。同时, 热稳定性和晶体结构得到了增强。流变实验显示, 随着CaCl2浓度增加, CCPS-3-1凝胶的表观黏度和存储模量也随之上升。Tanδ随CaCl2浓度的增加而减小, 且所有浓度下CCPS-3-1与CaCl2均能形成弱凝胶。此外, 静电相互作用在CCPS-3-1凝胶网络结构形成中起到了关键作用。结论 分离纯化得到的滁菊多糖CCPS-3-1经CaCl2诱导形成凝胶, 其凝胶机制可能为分子间氢键和Ca2+交联共同作用形成, 对滁菊多糖凝胶在食品、药品等领域的应用提供了理论依据。

, correspAuthors=陈春旭, authorNote=null, correspAuthorsNote=
* 陈春旭(1984—), 男, 博士, 副教授, 主要研究方向为食品分子加工与精准营养。E-mail:
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季文婷(1997—), 女, 硕士研究生, 主要研究方向为食品加工与安全。E-mail:

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季文婷(1997—), 女, 硕士研究生, 主要研究方向为食品加工与安全。E-mail:

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季文婷(1997—), 女, 硕士研究生, 主要研究方向为食品加工与安全。E-mail:

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注: A. 0% CaCl2、B. 0.5% CaCl2、C. 1.0% CaCl2、D. 1.5% CaCl2、E. 2.0% CaCl2、F. 2.5% CaCl2为放大倍数400; a. 0% CaCl2、b. 0.5% CaCl2、c. 1.0% CaCl2、d. 1.5% CaCl2、e. 2.0% CaCl2、f. 2.5% CaCl2; 放大倍数为1000。

, figureFileSmall=y7DnULqfDiH/Xk0+4VLHFQ==, figureFileBig=ajYA79ZmSfib8V3K1SaCCg==, tableContent=null), ArticleFig(id=1183428340485600038, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=EN, label=Fig.4, caption=Zeta potential and conductivity of CCPS-3-1 solution, figureFileSmall=rBKy0mqVzYwVe+hbx0r7Zg==, figureFileBig=vc4Svx8f0v8iYBQ/v0IksA==, tableContent=null), ArticleFig(id=1183428340540125991, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=CN, label=图4, caption=CCPS-3-1溶液的Zeta电位和电导率

注: 大写字母和小写字母分别表示Zeta电位和电导率值存在显着差异(P<0.05)。

, figureFileSmall=rBKy0mqVzYwVe+hbx0r7Zg==, figureFileBig=vc4Svx8f0v8iYBQ/v0IksA==, tableContent=null), ArticleFig(id=1183428340607234856, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=EN, label=Fig.5, caption=FTIR spectra of the lyophilized CCPS-3-1 gels, figureFileSmall=bSf32QUqbEj3mbyjuvkKMA==, figureFileBig=eIXncUUtwCLvHrUPFIw+Gw==, tableContent=null), ArticleFig(id=1183428340733063977, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=CN, label=图5, caption=冻干CCPS-3-1凝胶的FTIR光谱, figureFileSmall=bSf32QUqbEj3mbyjuvkKMA==, figureFileBig=eIXncUUtwCLvHrUPFIw+Gw==, tableContent=null), ArticleFig(id=1183428340800172842, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=EN, label=Fig.6, caption=XRD patterns of the lyophilized CCPS-3-1 gels, figureFileSmall=vT++MyRS7k8JzrBiHR1djw==, figureFileBig=w1/Da5aSgCTJuCFkg5mYqQ==, 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articleId=1153986653052986044, language=EN, label=Fig.8, caption=CCPS-3-1 gel LVR induced by different concentrations of CaCl2, figureFileSmall=ONDjkQBadHXU6Qut7QkHkg==, figureFileBig=TmYIOgPFUxCVnosSutbEZQ==, tableContent=null), ArticleFig(id=1183428341135717167, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=CN, label=图8, caption=不同浓度CaCl2诱导的CCPS-3-1凝胶LVR, figureFileSmall=ONDjkQBadHXU6Qut7QkHkg==, figureFileBig=TmYIOgPFUxCVnosSutbEZQ==, tableContent=null), ArticleFig(id=1183428341198631728, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=EN, label=Fig.9, caption=Static shear determination of CCPS-3-1 gel at different concentrations of CaCl2, figureFileSmall=NwSZGDYGOv/t46PIoqDGng==, figureFileBig=N5FAIOF9vQAiAHs+Vv1gVg==, tableContent=null), ArticleFig(id=1183428341269934897, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986653052986044, language=CN, 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滁菊多糖CCPS-3-1凝胶制备及其特性研究
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季文婷 1 , 韩天祥 1 , 徐友梅 2 , 刘明 3 , 陈春旭 1, 2, *
食品安全质量检测学报 | 本期专题：功能性食品与功能性成分 2025,16(3): 130-138
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食品安全质量检测学报 | 本期专题：功能性食品与功能性成分 2025, 16(3): 130-138
滁菊多糖CCPS-3-1凝胶制备及其特性研究
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季文婷1 , 韩天祥1, 徐友梅2, 刘明3, 陈春旭1, 2, *
作者信息
  • 1.安徽科技学院食品工程学院, 凤阳 233100
  • 2.安徽省功能农业与功能食品重点实验室, 滁州 233100
  • 3.凤阳临淮油脂有限公司, 凤阳 233100
  • 4.安徽欧奈生物科技有限公司, 滁州 239000

    • 季文婷(1997—), 女, 硕士研究生, 主要研究方向为食品加工与安全。E-mail:

通讯作者:

* 陈春旭(1984—), 男, 博士, 副教授, 主要研究方向为食品分子加工与精准营养。E-mail:
Preparation and characterization of gel from Chuzhou chrysanthemum polysaccharide CCPS-3-1
Wen-Ting JI1 , Tian-Xiang HAN1, You-Mei XU2, Ming LIU3, Chun-Xu CHEN1, 2, *
Affiliations
  • 1. College of Food Engineering, Anhui Science and Technology University, Fengyang 233100, China
  • 2. Anhui Provincial Key Laboratory of Functional Agriculture and Functional Foods, Chuzhou 233100, China
  • 3. Fengyang Linhuai Oils Co., Ltd., Fengyang 233100, China
  • 4. Anhui Ounai Biotechnology Co., Ltd., Chuzhou 239000, China
出版时间: 2025-02-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241111006
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摘要
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目的 制备滁菊多糖CCPS-3-1凝胶, 并研究其相关特性。方法 以滁菊为原料, 分离纯化得到酸性多糖CCPS-3-1, 与CaCl2溶液混合, 通过离子交联作用形成凝胶。运用扫描电子显微镜、傅里叶红外光谱、X-射线衍射、差示扫描量热法等测定方法对其形成的凝胶特性进行全面分析, 并用尿素和乙二胺四乙酸处理进一步阐明其胶凝力和胶凝机制。结果 在pH 6.0~8.0范围内, CCPS-3-1溶液相对稳定。经CaCl2诱导后, CCPS-3-1凝胶呈现出良好的透明度, 有一定的硬度和良好的稳定性, 凝胶网络结构致密, 表面光滑且均匀。同时, 热稳定性和晶体结构得到了增强。流变实验显示, 随着CaCl2浓度增加, CCPS-3-1凝胶的表观黏度和存储模量也随之上升。Tanδ随CaCl2浓度的增加而减小, 且所有浓度下CCPS-3-1与CaCl2均能形成弱凝胶。此外, 静电相互作用在CCPS-3-1凝胶网络结构形成中起到了关键作用。结论 分离纯化得到的滁菊多糖CCPS-3-1经CaCl2诱导形成凝胶, 其凝胶机制可能为分子间氢键和Ca2+交联共同作用形成, 对滁菊多糖凝胶在食品、药品等领域的应用提供了理论依据。

滁菊多糖  /  凝胶  /  凝胶特性

Objective To prepare Chuzhou chrysanthemum polysaccharide CCPS-3-1 gel and investigate its relevant properties. Methods Acidic polysaccharide CCPS-3-1 was isolated and purified from Chuzhou chrysanthemum and mixed with CaCl2 solution to form a gel via ionic crosslinking. The gel's characteristics were comprehensively analyzed using scanning electron microscopy, Fourier-transform infrared spectroscopy, X-ray diffraction, and differential scanning calorimetry. Additionally, treatment with urea and ethylenediaminetetraacetic acid was employed to further elucidate its gel strength and gelling mechanism. Results CCPS-3-1 solution was relatively stable within a pH range of 6.0-8.0. Upon induction by CaCl2, the CCPS-3-1 gel exhibited good transparency, moderate hardness, and high stability. The gel network had a dense, smooth, and uniform structure. Moreover, the thermal stability and crystalline structure were enhanced. Rheological experiments revealed that as the CaCl2 concentration increased, the apparent viscosity and storage modulus of the CCPS-3-1 gel also increased. In addition, the loss tangent (Tanδ) decreased with increasing CaCl2 concentration, indicating the formation of weak gels at all tested concentrations. Furthermore, electrostatic interactions played a crucial role in the formation of the gel network structure. Conclusion The Chuzhou chrysanthemum polysaccharide CCPS-3-1, isolated and purified in this study, forms a gel upon CaCl2 induction. Its gelling mechanism likely involves the synergistic effects of intermolecular hydrogen bonding and Ca2+ crosslinking. These findings provide a theoretical foundation for the application of CCPS-3-1 gel in food, pharmaceutical, and other fields.

Chuzhou chrysanthemum polysaccharide  /  gel  /  gel properties
季文婷, 韩天祥, 徐友梅, 刘明, 陈春旭. 滁菊多糖CCPS-3-1凝胶制备及其特性研究. 食品安全质量检测学报, 2025 , 16 (3) : 130 -138 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241111006
Wen-Ting JI, Tian-Xiang HAN, You-Mei XU, Ming LIU, Chun-Xu CHEN. Preparation and characterization of gel from Chuzhou chrysanthemum polysaccharide CCPS-3-1[J]. Journal of Food Safety & Quality, 2025 , 16 (3) : 130 -138 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241111006
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0 引言
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近年来, 天然多糖因其多样的生物活性受到关注, 部分植物多糖以增稠、胶凝和乳化等功能特性成为重要天然食品添加剂, 用于改善食品质地和稳定性[1-2]。这些多糖的亲水特性和流变特性是它们在食品和制药行业中越来越多地用作胶凝剂、增稠剂和乳化剂的主要原因[3]。多糖凝胶形成涉及氢键、静电和离子等相互作用[4]。带电多糖在凝胶化中常通过静电作用聚集, 而盐离子的加入能屏蔽这些静电, 帮助评估静电作用。此外, 盐离子对构建阴离子聚合物线性结构至关重要, 如钙离子(Ca2+)与果胶形成钙桥, 是凝胶化的关键[5]。
滁菊(Chuzhou chrysanthemum), 作为我国知名的药用菊花, 蕴含丰富的黄酮类和挥发油等化学成分, 已证实具备抗氧化、抗炎和降血脂等药理作用。尽管如此, 对于滁菊多糖(Chuzhou chrysanthemum polysaccharides, CCPS)及其凝胶特性的研究尚不充分, 尤其是在多糖凝胶领域的潜在应用价值尚未被完全开发。
本研究通过分离纯化CCPS组分CCPS-3-1, 利用不同浓度的CaCl2诱导其形成凝胶。旨在深入探究滁菊多糖CCPS-3-1凝胶的特性, 包括其形成机制、流变学、微观结构以及热稳定性等, 这不仅有助于深化其结构与功能关系的认识, 而且有望为开发以CCPS凝胶为基础的新型产品提供科学支撑, 进一步推动滁菊资源的高值化利用。
1 材料与方法
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1.1 材料与试剂
滁菊购自滁州市宏福保健品有限公司。
苯酚、氯化钙(CaCl2)、尿素、氢氧化钠(分析纯)、盐酸(HCl)、硫酸(纯度98%)(国药集团化学试剂有限公司); 快流速二乙氨基乙基—琼脂糖凝胶层析柱(DEAE-Sepharose Fast Flow, 美国GE医疗生命科学部); 葡聚糖凝胶填料(Sephadex G-100, 美国GE公司); 溴化钾(分析纯, 上海阿拉丁试剂有限公司); 乙二胺四乙酸(ethylenediaminetetraacetic acid, EDTA)(分析纯, 上海麦克林生化科技有限公司);。
1.2 仪器与设备
FA2004A电子天平(精度0.1 mg, 上海精天电子仪器有限公司); Bettersize2600激光粒度分析仪(丹东百特仪器有限公司); TGA4000热重分析仪(美国PE公司); SU8020扫描电镜(日本日立公司); D8 Advance X射线衍射仪(德国布鲁克公司); Nicolet iS10傅里叶红外光谱仪(美国Thermo Fisher Scientific公司); Q600 SDT热重分析仪(美国TA公司)。
1.3 实验方法
1.3.1 滁菊多糖的分离纯化及成分分析
(1)滁菊多糖CCPS-3-1的分离纯化
滁菊粗多糖的提取参考韩天祥等[6]的研究方法。首先用80%的乙醇水溶液提取两次干燥滁菊粉末, 以去除可溶性糖、色素和其他小分子。沉淀烘干后按料液比1:25 (g/mL)加入蒸馏水, 85 ℃条件下热水浸提3次, 每次浸提2 h, 过滤、离心并合并上清液。在45 ℃下旋转蒸发浓缩提取液, 浓缩至体积为原提取液体积的1/3~1/5且溶液出现较为明显的黏度增加的现象。接着向浓缩液加入4倍体积的无水乙醇, 混合均匀, 并在4 ℃下保持12 h以沉淀多糖。通过氯仿-正丁醇法对沉淀多糖去除蛋白, 之后透析, 然后冻干后得到滁菊粗多糖CCPS。将CCPS溶液(20 mg/mL)加载到DEAE-Sepharose Fast Flow的凝胶柱上(2.6 cm×50 cm), 分别使用蒸馏水、NaCl (0.1、0.3和0.5 mol/L)溶液以60 mL/h洗脱2个柱体积。随后, 得到CCPS-1、CCPS-2和CCPS-3 3个组分。CCPS-3在Sephadex G-100色谱柱上进一步以12 mL/h的流速以蒸馏水洗脱, 得到单一组分CCPS-3-1。
(2)滁菊多糖CCPS-3-1的成分分析
利用苯酚硫酸法[7]、考马斯亮蓝法[8]、间羟联苯法[9]、和福林酚法[10]分别测定CCPS-3-1总糖、蛋白质、糖醛酸和多酚含量。
总糖含量测定如下: 首先, 将烘干的葡萄糖标准品配制成100 μg/mL的葡萄糖标准溶液。接着, 分别取0.0、0.2、0.4、0.6和0.8 mL的标准溶液置于试管中, 再用蒸馏水分别补至1.0 mL。然后依次向每个试管中加入0.5 mL的6%苯酚溶液和2.5 mL的浓硫酸, 摇匀后冷却, 在490 nm处测定吸光值, 依据吸光值与相应葡萄糖浓度绘制标准曲线。接着配制50 μg/mL的CCPS-3-1样品溶液, 取0.5 mL加入试管中, 然后依次加入0.5 mL的蒸馏水、0.5 mL的6%苯酚溶液和2.5 mL的浓硫酸, 摇匀后冷却, 在490 nm处测定其吸光值, 最后根据标准曲线计算总糖含量, 实验重复3次。
蛋白质含量测定如下: 首先, 将200 mL的考马斯亮蓝G-250溶解于100 mL 90%乙醇中, 然后向其中加入200 mL 85%磷酸溶液, 最后用蒸馏水定容至2000 mL。将混合溶液过滤后保存在棕色瓶中备用。接着, 配制50 μg/mL牛血清白蛋白(bovine serum albumin, BSA)标准溶液, 取0.0、0.2、0.4、0.6和0.8 mL的BSA标准溶液至试管中, 加入蒸馏水至1.0 mL, 涡旋均匀后, 加入5.0 mL的考马斯亮蓝溶液, 混匀后静置5 min, 然后在595 nm处测定吸光值。根据吸光值与相应BSA浓度绘制标准曲线。接着, 配制5 mg/mL的CCPS-3-1样品溶液, 取0.5 mL于试管内, 加入蒸馏水至1.0 mL, 按照上述标准曲线绘制过程中的操作步骤测定595 nm的吸光值, 最后根据标准曲线计算蛋白质含量。
糖醛酸含量测定如下: 首先, 配制60 g/mL的半乳糖醛酸标准溶液, 分别取0.00、0.05、0.10、0.15、0.20 mL置于5 mL的离心管中, 用蒸馏水补足至0.25 mL。将离心管放入冰浴中预冷, 然后向每个管中加入1.5 mL四硼酸钠-硫酸溶液(浓度为0.0125 mL/L), 混合均匀, 置于沸水浴中5 min, 快速将离心管放入冰浴中冷却至室温, 加入25 mL间羟联苯溶液(以0.5% NaOH配制的0.15%间羟联苯溶液), 混合均匀后, 在520 nm处测定各管吸光值, 并绘制标准曲线。接着, 配制160 μg/mL的CCPS-3-1样品溶液, 随后取250 μL按照上述半乳糖醛酸标准溶液的测定方法测定其吸光值, 并根据标准曲线计算糖醛酸含量。
多酚含量测定如下: 首先, 将福林酚溶液需稀释10倍作为工作液, 每100 mL蒸馏水中溶解10 mg的碳酸钠以制备10%的碳酸钠溶液; 称取10 mg没食子酸, 加入去离子水并定容至100 mL以配制没食子酸标准溶液。接着, 分别吸取没食子酸标准溶液0.05、0.10、0.15、0.20、0.25 mL至试管中, 每管去离子水补至0.50 mL后加入1 mL福林酚工作液, 混匀后于30 ℃避光条件反应5 min, 随后每管再加入2 mL 10%碳酸钠溶液, 混匀后继续于30 ℃避光下反应1 h, 反应结束后在747 nm处测定吸光值。以没食子酸标准溶液浓度为横坐标, 吸光值为纵坐标, 绘制标准曲线。取1 mL 1 mg/mL的CCPS-3-1, 按照标准曲线的测定方法测定吸光值, 将测得的吸光值代入没食子酸标准曲线, 计算得到CCPS-3-1中总多酚的含量, 以没食子酸当量(gallic acid equivalent, GAE)表示。
1.3.2 CCPS-3-1凝胶的制备
CCPS-3-1凝胶的制作参考LUO等[11]报道的方法, 并作适当的修改。在室温下, 首先称取2 g干燥粉碎的CCPS-3-1于200 mL去离子水中, 搅拌均匀后, 将混合溶液于80 ℃下加热2 h, 边加热边搅拌, 即可得到CCPS-3-1储备溶液。同时, 在室温下分别将不同量的CaCl2溶解于25 mL去离子水中, 制备成不同浓度(0%、0.5%、1.0%、1.5%、2.0%、2.5%)的CaCl2储备溶液。接着, 在室温下将0.5 mL CaCl2溶液与2 mL CCPS-3-1溶液持续搅动混溶, 最后将混合物溶液冷却至4 ℃促使CaCl2与CCPS-3-1混合液中的成分交联, 让液体混合液逐渐变成固体凝胶状态。
1.3.3 扫描电子显微镜观察
扫描电子显微镜(scanning electron microscopy, SEM)是一种利用高能电子束来观察和分析样品表面的微观结构和组成的技术, 可以通过SEM观察CCPS-3-1凝胶的微观结构。首先将制备好的CCPS-3-1凝胶立即浸入液氮中快速冷冻, 然后在真空冷冻干燥机中-50 ℃脱水48 h。接着将冻干凝胶断裂, 并在其表面涂上一层金粉, 最后在高真空模式下进行SEM观察, 加速电压为5 kV[12]。
1.3.4 Zeta电位测量
CCPS-3-1的Zeta电位和电导率采用动态光散射通过应用波长633 nm的激光源进行测量[13]。将0.01 g CCPS-3-1在25 ℃的去离子水中分散2 h, 以制备CCPS-3-1分散液(质量浓度为0.01 g/100 mL)。使用0.1 mol/L HCl和0.01 mol/L NaOH将分散溶液的pH调节至3.0~8.0, 测量温度为25 ℃, 平衡时间为120 s, 折射率为1.33。
1.3.5 傅里叶红外光谱测定
傅里叶红外光谱(Fourier transform infrared spectroscopy, FTIR)实验是将3 mg CCPS-3-1和200 mg溴化钾粉末混合, 用研钵和研杵将混合物捣成粉末状。用压片机将混合物压成1 mm的薄片, 并在红外光谱仪上扫描。波长范围为4000~400 cm-1。
1.3.6 X射线衍射测定
X-射线衍射(X-ray diffraction, XRD)实验使用了配备CuKα灯和镍滤光片的XRD仪器。首先将冻干的CCPS-3-1凝胶研磨成粉末形式。然后记录CCPS-3-1在5°到55°之间的XRD谱图, 采用0.02°的步长, 并将扫描速度设定为4°/min。
1.3.7 差式扫描量热法测定
差式扫描量热法(different scanning calorimrtry, DSC)使用热分析设备。以10 ℃/min的线性加热速率将约10 mg冻干CCPS-3-1凝胶从室温加热至500 ℃。以纯氮气为吹扫气体, 空铝盘作为参比[14]。
1.3.8 凝胶的流变性能测定
采用MCR 302流变仪和平行板传感器(间隙0.5 mm, 直径40 mm)测定CCPS-3-1凝胶的流变学分析, 并作适当修改[15]。在每次测量前所有样品均需平衡2 min, 然后将不同浓度的CaCl2 (0.5%、1.0%、1.5%、2.0%、2.5%)的CCPS-3-1 (1%)溶解在蒸馏水中。
(1)线性黏弹区
通过记录25 ℃下, 频率为1 Hz, 振荡应变范围从0.1%到100.0%的所有样品的存储模量(Gʹ), 以确定线性黏弹区(linear viscoelastic region, LVR), 并选择合适形变量进行后续动态黏弹性测定。
(2)表观黏度测定
在25 ℃和剪切速率为0.01~100.00 s-1条件下测定凝胶的表观黏度随剪切速率的变化。每个样品将进行3次平行测定, 测定结果取平均值。
(3)动态黏弹性测定
在25 ℃条件下, 通过在0.1~10.0 Hz测定凝胶的存储模量(Gʹ)、损耗模量(Gʺ)和损耗因子(Tanδ=Gʺ/Gʹ)随频率的变化情况, 其形变量由LVR测定结果决定。
1.3.9 凝胶力
尿素和EDTA实验广泛用于评估多糖凝胶中的胶凝力。首先, 将尿素(1.0 g和2.5 g)和EDTA (0.05 g和0.10 g)分别溶解在10 mL 1.5%的CaCl2溶液中。然后, 将40 mL 1%的CCPS-3-1溶液与CaCl2溶液充分搅拌混合。最终溶液中, 尿素的浓度分别是2.0%和5.0%, EDTA的浓度分别是0.1%和0.2%。最后将混合溶液冷却至4 ℃, 并在该温度下保存12 h。同时, 以不添加尿素和EDTA的CCPS-3-1凝胶作为对照[11]。
1.4 数据处理
所有实验结果重复3次, 均以平均值±标准偏差的形式呈现。为了评估结果的显著性, 通过单因素方差分析(analysis of variance, ANOVA)和IBM SPSS Statistics 20软件中邓肯检验来评估组间的差异。如果P<0.05, 则认为存在显著差异。
2 结果与分析
收起
2.1 CCPS-3-1的提取和纯化
滁菊经热水浸提后, 再经脱蛋白和乙醇沉淀等步骤得到粗多糖组分CCPS, 提取率达7.15%±0.26%。CCPS经过DEAE Sepharose Fast Flow层析柱纯化, 得到3个多糖组分CCPS-1、CCPS-2、CCPS-3(图1A)。选择CCPS-3组分加载到Sephadex G-100层析柱, 并以蒸馏水作为洗脱试剂进一步纯化, 得到酸性多糖CCPS-3-1(图1B)。
2.2 CCPS-3-1的化学成分分析
CCPS-3-1含有30.03%±0.87%的总糖, 表明通过热水浸提的滁菊多糖组分CCPS-3-1具有很高的糖含量, 是其主要组成部分之一; 含有56.97%±0.94%的糖醛酸, 说明CCPS-3-1是一种酸性多糖, 而糖醛酸通常参与分子间的交联, CCPS-3-1较高的糖醛酸比例可能与多糖的某些生物学特性(如凝胶形成能力)相关; 蛋白质含量为0.26%±0.24%, 表明蛋白质不是CCPS-3-1的主要成分, 低蛋白质含量可能有助于减少非特异性反应和提高纯度; 多酚含量则为(0.46±0.06) mg GAE/100 mg, 含量相对较低, 不是CCPS-3-1的主要成分, 这可能与多糖提取过程中加入乙醇有关, 乙醇具有吸附多酚类物质的性质。
2.3 CCPS-3-1凝胶形态学分析
不同浓度的CaCl2对CCPS-3-1的凝胶化诱导作用如图2所示, 在Ca2+离子存在时, CCPS-3-1溶液可以形成自支撑的稳定凝胶。以1% CCPS-3-1水溶液为基础, 加入不同量的CaCl2制备凝胶。当CaCl2的添加量为0%时, CCPS-3-1水溶液无法形成凝胶, 说明CaCl2是形成CCPS-3-1凝胶网络的先决条件, CaCl2通过提供Ca2+离子来促进CCPS-3-1之间的交联。随着CaCl2添加量的增加, 凝胶的交联密度和饱和度逐渐增加, 表现为凝胶的色泽变深。然而, 随着CaCl2浓度进一步增加, 凝胶的硬度和稳定性反而降低, 凝胶变得脆弱且不均匀, 凝胶的透明度降低变得更加浑浊, 表现为凝胶的形状变得松散和不规则。这可能是由于CaCl2过量导致CCPS-3-1分子链的缠绕和破坏, 从而影响凝胶的空隙结构和水分保持能力。
SEM技术在研究多糖凝胶的微观结构中发挥着重要作用, 可以提供高分辨率的表面形貌图像, 揭示多糖凝胶的表面特征, 如表面粗糙度、孔隙结构等, 有助于评估凝胶的表面性质和形貌[16]。图3是不同浓度的CaCl2影响CCPS-3-1的SEM图像。可以观察到, 当CaCl2的添加量为0%时, 凝胶表现出更加均匀和光滑的表面和更小的孔隙, 并且随着CaCl2浓度增加到1.5%, 结构变得更加致密。当CaCl2浓度达到2.5%时, 凝胶的网络结构表现出更加不均匀、疏松、粗糙的表面和更大的孔隙。这可能是由于钙离子浓度过高导致凝胶过于坚硬, 影响其弹性和可塑性, 使光滑的表面变得粗糙, 并且由于正负离子的相互作用, 从而使凝胶结构产生更大的孔隙。SEM图像结果与凝胶外观的分析结果相符。
2.4 CCPS-3-1的Zeta电位分析
Zeta电位是表征凝胶分散系稳定性的重要指标, 它反映了分散离子表面与周围介质之间的电势差。由图4可知, CCPS-3-1在pH 3.0~8.0时具有负的Zeta电位, 这与半乳糖醛酸单元-COOH在酸性条件下解离有关。当pH从3.0增加到6.0时, Zeta电位从-12 mV变化到-35 mV。说明溶液中多糖分子之间的静电斥力较强, 能够有效阻止分子间的相互聚集, 使溶液处于相对稳定的状态。CCPS-3-1的Zeta电位在pH 6.0~8.0范围内相对稳定。这一结果与有关报道中一种商品果胶相似[17], 可能是由于滁菊多糖和果胶在单糖组成和结构上具有一定的相似性。CCPS-3-1中高水平的负电荷使其能够与阳离子相互作用并迅速聚集, 这是CCPS-3-1有凝胶特性的重要原因之一[18]。正如有关研究表明, 当多糖分子作为阴离子或阳离子存在时, 它们可以通过相互作用与带相反电荷的其他分子结合, 这种结合可以增加凝胶网络结构的稳定性, 提高凝胶的机械强度和弹性[19-21]。
当多糖溶液通过交联固化成凝胶时, 其内部的微观结构发生变化, 影响离子的迁移路径和速度, 进而影响电导率。CCPS-3-1溶液的电导率为0.0495~0.0775 mS/cm (pH 4.0~8.0), 与芒果皮多糖的电导率相近[13], 表明CCPS-3-1溶液具有良好的稳定性和网络结构。
2.5 CCPS-3-1凝胶的FTIR分析
FTIR分析可以为多糖凝胶的研究提供丰富的结构和成分信息, 有助于深入理解多糖凝胶的形成机制、性质和应用。红外光谱中峰的位置、强度及形状的变化可以反映分子间的相互作用。如图5所示, 单独CCPS-3-1 (CaCl2添加量为0%)在3409 cm-1处表现出明显的吸收峰, 这归因于糖类分子中的O-H伸缩振动; 在1607 cm-1处的吸收峰对应于糖醛酸中的羰基C=O振动, 这也同样表明滁菊多糖CCPS-3-1组分是酸性多糖。随着CaCl2浓度的增加, 吸收带向较低波数移动。在CaCl2浓度为1.0%时, 吸收峰达到3378 cm-1, 表明谱带发生显著红移。这可能是由于Ca2+会和CCPS分子中的官能团发生相互作用, 这种相互作用使CCPS分子链间的氢键等作用力发生变化, 特别是通过改变多糖分子上的羟基之间的氢键强度, 导致多糖分子的振动状态改变。当CaCl2浓度增加到2.5%时, 谱带向更高波数移动, 吸收峰中心位于3428 cm-1。研究结果表明, 低浓度的CaCl2添加促进了分子间氢键的形成, 但过量的CaCl2会削弱分子间氢键[14,22]。
2.6 CCPS-3-1凝胶的XRD分析
不同CaCl2浓度制备得到的CCPS-3-1凝胶的XRD谱图如图6所示, 多糖的衍射峰强度与CaCl2的浓度有关。结果表明, 单独的CCPS-3-1无尖锐峰, 在20°处有小的部分结晶带, 说明CCPS-3-1为非晶体无定形材料, 与白及多糖水凝胶的XRD分析结果相一致[23]。随着CaCl2浓度的增加, 其结晶度也增加, 在约32°和45°处出现了新的衍射峰。这说明Ca2+与CCPS-3-1之间存在相互作用, 形成了稳定的配合物, 增强了分子间的氢键和疏水相互作用。当Ca2+浓度为1.5%时, CCPS-3-1凝胶的结晶度达到最大, 而更高浓度的CaCl2则导致结晶颗粒的减少和分散。因此, 特征峰的强度减小反映了多糖凝胶结晶度的变化, 表明CCPS-3-1凝胶具备稳定的结构。
2.7 CCPS-3-1凝胶的DSC分析
DSC是一种常用的热分析技术, 可以测量物质在升温、降温或恒温过程中吸收或释放的能量。对于多糖类物质, DSC通过观察多糖在加热过程中的分解温度或吸热峰, 评估多糖的热稳定性[24]。如图7 DSC结果表明, 单独CCPS-3-1(未添加CaCl2)的吸热过程发生在157 ℃附近, 放热过程发生在248 ℃附近。随着CaCl2浓度的增加, CCPS-3-1凝胶的放热峰值逐步增加, 这一现象与CaCl2的浓度有关, 随着CaCl2浓度的增加, 可能发生了更强烈的CCPS-3-1和CaCl2之间的相互作用, 更多的Ca2+参与到交联过程中, 使得CCPS-3-1凝胶结构更为致密和稳定, 从而提高了CCPS-3-1凝胶的热稳定性。表明CaCl2的加入可能会提高CCPS-3-1凝胶的熔融温度, 表明需要更多的能量来破坏凝胶结构, 从而抵抗热诱导的解体。
2.8 CCPS-3-1凝胶的流变学分析
2.8.1 LVR分析
对多糖凝胶进行LVR分析, 能够深入了解多糖凝胶在不同应变条件下的黏弹性特征, 进一步揭示其结构稳定性、交联程度等。不同浓度的CaCl2诱导的CCPS-3-1凝胶LVR如图8所示, CCPS-3-1凝胶在应力达到一定值后, 存储模量开始下降, 说明了CCPS-3-1分子链发生了解聚, 交联密度下降, 并且随着CaCl2浓度的增加, CCPS-3-1凝胶的形成能力可能发生变化。随着CaCl2浓度的升高, CCPS-3-1凝胶的存储模量也随着升高, LVR有拓宽趋势, 表明高浓度CaCl2诱导的凝胶能承受更大的形变而不破坏其结构, 表明CaCl2在凝胶网络的形成中起到了关键作用, CaCl2浓度的增加可能会增强CCPS-3-1凝胶的稳定性和强度。
2.8.2 静态剪切测定结果分析
静态剪切测定可以用来评估多糖在外力作用下的流动特性。图9A是CCPS-3-1凝胶的剪切应力变化图, 结果表明, 在不同CaCl2浓度下, CCPS-3-1凝胶的剪切应力都随着剪切速率的增加而上升, 并呈现出非线性特征。这表明CCPS-3-1凝胶在各种CaCl2浓度条件下都表现出非牛顿流体的假塑性特性[25]。这可能是由于多糖分子之间的相互作用, 如氢键和疏水作用, 在剪切力的作用下可能会减弱, 使得分子间的束缚力降低, 凝胶在受力时更容易流动, 从而表现出假塑性特性[25]。图9B是CCPS-3-1凝胶的表观黏度变化图, 可以观察到, 在所有不同CaCl2浓度下, CCPS-3-1凝胶的黏度随着剪切速率的增加而降低, 这直接反映了CCPS-3-1凝胶剪切稀化现象[26-27], 与大豆种皮多糖添加量对乳液凝胶表观黏度随剪切速率变化的结果相符[28]。此外, 相同剪切速率下, 随着CaCl2浓度的增加, CCPS-3-1凝胶的黏度也随之增加, 抗剪切性增强。
2.8.3 动态频率扫描结果
不同浓度CaCl2对CCPS-3-1凝胶黏弹性行为的影响如图10A~C所示, 在0.1~10.0 Hz的频率扫描范围内, CCPS-3-1凝胶的Gʹ和Gʺ值随Ca2+浓度从0.5%增加到2.5%而增加。此外, Gʹ的值始终大于Gʺ, 表明所有样品此时可以形成凝胶结构从而限制水分子的迁移。对于弹性行为, Gʹ>Gʺ (Tanδ<1), 而Gʹ<Gʺ (Tanδ>1)是黏性行为的特征[29-30]。
上述现象也可以Tanδ来解释。如图10C所示, 所有样品的Tanδ值均随频率的增加而增加。Tanδ值小于1, 说明CCPS-3-1样品在不同Ca2+浓度下表现出固体弹性, 且表现出较弱的凝胶行为。这一现象表明, 静电相互作用在CCPS-3-1凝胶网络结构的形成中起到了重要的作用。之前的研究也报告了类似的结果。例如, 来自Mesona blumes的多糖也表明, 当Ca2+的加入超过一定点时, Tanδ的值增加[4]。适当浓度的Ca2+离子增加了静电相互作用, 从而增强了CCPS-3-1凝胶的流变行为。
2.8.4 尿素和EDTA实验分析
尿素和EDTA被用于评估植物多糖的胶凝机制, 发现尿素和EDTA的加入会显著改变凝胶的功能特性[27]。尿素作为一种变性剂, 可能会改变多糖的三维结构, 从而影响其凝胶特性。EDTA作为一种螯合剂, 能够与金属离子形成稳定的复合物, 可能会影响依赖于金属离子(如Ca2+)来维持结构的植物凝胶。如图11所示, 添加2.0%尿素后, CCPS-3-1胶凝力明显减弱, 添加5.0%尿素则完全消除了胶凝作用, 这可能是由于尿素破坏了多糖分子间的氢键。当添加0.1%或0.2% EDTA时, 没有形成凝胶, 这可能是因为EDTA螯合了多糖分子中的金属离子, 切断了CCPS-3-1分子间通过Ca2+形成的交联键, 从而影响了CCPS-3-1凝胶的形成和维持。这一发现表明氢键和Ca2+交联应同时促进CCPS-3-1凝胶化。
3 讨论与结论
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以滁菊为原材料, 分离纯化得到CCPS-3-1组分, CCPS-3-1的主要成分为酸性多糖。多糖的化学组成对其凝胶特性有着直接的影响, 其凝胶能力与其分子间的相互作用有关, 这些相互作用包括氢键、疏水作用、离子键和分子间的堆积作用力等。多糖分子中的羟基可以通过氢键形成稳定的分子间交联, 从而在冷却或遇水时形成凝胶, 或者官能团水解(如乙酰基团)可以通过与多糖分子相互作用, 增强凝胶网络的稳定性[31-32]。
通过不同浓度CaCl2诱导CCPS-3-1形成凝胶, 酸性糖的存在使CCPS-3-1对Ca2+离子具有敏感性; 同时, 高含量的糖醛酸可能表明CCPS-3-1具有高支化结构, 从而有利于通过链缠结形成分子间氢键。当适量的Ca2+离子存在下, CCPS-3-1在4 ℃低温下表现出显著的胶凝能力; 然而, 过量的Ca2+会导致凝胶作用减弱或消失。经CaCl2诱导后, CCPS-3-1的热稳定性得到提升, 晶体结构更加稳定。流变实验结果表明, CCPS-3-1凝胶的表观黏度和存储模量随着CaCl2浓度的增加而增大, 而Tanδ随着CaCl2浓度的增加而减小。此外, 静电相互作用在CCPS-3-1凝胶网络结构的形成中起到了重要的作用。综合分析, CCPS-3-1的凝胶机制涉及分子间氢键和Ca2+交联的同时形成。研究结果可为滁菊多糖在凝胶领域的应用提供重要指导。
基金
收起
  • 国家自然科学基金项目(31901617)
  • 安徽省教育厅拔尖人才项目(gxyqZD2021125)
  • 安徽科技学院优秀人才项目(XJYXRC202101)
  • 安徽科技学院稳定人才项目(SPWD202101)
  • 安徽省教育厅自然科学研究基金项目(2022AH040236)
  • 安徽省大学生创新创业训练计划项目(S202310879232)
  • 安徽省学科带头人培养计划项目(DTR2024034)
  • 安徽省青年拔尖人才学者计划项目
文献
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241111006
  • 接收时间：2024-11-11
  • 首发时间：2025-07-21
  • 出版时间：2025-02-15
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  • 收稿日期：2024-11-11
基金
国家自然科学基金项目(31901617)
安徽省教育厅拔尖人才项目(gxyqZD2021125)
安徽科技学院优秀人才项目(XJYXRC202101)
安徽科技学院稳定人才项目(SPWD202101)
安徽省教育厅自然科学研究基金项目(2022AH040236)
安徽省大学生创新创业训练计划项目(S202310879232)
安徽省学科带头人培养计划项目(DTR2024034)
安徽省青年拔尖人才学者计划项目
作者信息
    1.安徽科技学院食品工程学院, 凤阳 233100
    2.安徽省功能农业与功能食品重点实验室, 滁州 233100
    3.凤阳临淮油脂有限公司, 凤阳 233100
    4.安徽欧奈生物科技有限公司, 滁州 239000

通讯作者:

* 陈春旭(1984—), 男, 博士, 副教授, 主要研究方向为食品分子加工与精准营养。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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