Article(id=1151437191147778672, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151437189243089177, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250228007, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1740672000000, receivedDateStr=2025-02-28, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752453619037, onlineDateStr=2025-07-14, pubDate=1749916800000, pubDateStr=2025-06-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752453619037, onlineIssueDateStr=2025-07-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752453619037, creator=13701087609, updateTime=1752453619037, updator=13701087609, issue=Issue{id=1151437189243089177, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='11', pageStart='1', pageEnd='320', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752453618584, creator=13701087609, updateTime=1767768054466, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1215670588966883492, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151437189243089177, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1215670588966883493, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151437189243089177, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=307, endPage=313, ext={EN=ArticleExt(id=1151895327738524299, articleId=1151437191147778672, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Optimization of ultrasonic extraction process of proanthocyanidins from Polygonum aviculare by response surface method, columnId=1166765750476878043, journalTitle=Journal of Food Safety & Quality, columnName=Original article, runingTitle=null, highlight=null, articleAbstract=

Objective To optimize the ultrasonic-assisted extraction process of proanthocyanidins from Polygonum aviculare by response surface method, to obtain the best extraction conditions. Methods The main factors affecting the extraction amount were investigated through single-factor experiments. Based on the results of the single-factor experiments, the Box-Behnken design and response surface methodology were used to optimize the single factors, including the volume fraction of ethanol, ultrasonic time, and solid-liquid ratio, which had an impact on the extraction amount of proanthocyanidins from Polygonum aviculare. The optimal extraction process conditions were obtained. Results The optimized results of the optimal process conditions for ultrasonic-assisted extraction of proanthocyanidins from Polygonum aviculare were as follows: The ultrasonic time was 34 min, the volume fraction of ethanol was 65%, and the solid-liquid ratio was 1:46 (g:mL). Under these process conditions, the final extraction amount of proanthocyanidins reached 2.18%, which was consistent with the model validation value of 2.12%. Conclusion This study shows that the process condition parameters optimized by the response surface methodology are effective and feasible. The response surface methodology is successfully applied to optimize the extraction parameters of ultrasonic-assisted extraction of proanthocyanidins from Polygonum aviculare, providing a research basis for the later promotion and application of proanthocyanidins from Polygonum aviculare.

, correspAuthors=Qi SHANG, 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=Qi SHANG, Yu SONG, Yu LIU, Jian-Ping WANG, Ming-Jun DAI), CN=ArticleExt(id=1151895343874011363, articleId=1151437191147778672, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=响应面法优化超声提取萹蓄原花青素工艺, columnId=1151895322281734685, journalTitle=食品安全质量检测学报, columnName=食品加工与工艺, runingTitle=null, highlight=null, articleAbstract=

目的 通过利用响应面法优化超声辅助提取食药植物萹蓄中原花青素的工艺, 获取最佳工艺条件。方法 通过单因素试验考察影响提取量的主要因素, 在单因素试验结果的基础上, 用Box-Behnken设计及响应面分析法, 对萹蓄中原花青素提取量的影响单因素乙醇体积分数、超声时间、料液比进行优化, 获取最佳提取工艺条件。结果 萹蓄原花青素超声辅助提取最佳工艺条件优化结果分别为: 超声时间34 min、乙醇体积分数65%、料液比1:46 (g:mL), 在此工艺条件下, 原花青素的最终提取量达到2.18%, 与模型验证值2.12%相符合。结论 本研究表明响应面法优化得出的工艺条件参数是有效可行的, 成功地应用响应面法优化了超声辅助提取萹蓄中原花青素的提取参数, 为萹蓄原花青素的后期推广应用提供研究基础。

, correspAuthors=尚琪, authorNote=null, correspAuthorsNote=
* 尚琪(1990—), 男, 硕士, 工程师, 主要研究方向为食品及相关产品的检验检测, 药食同源植物天然产物的提取与分离及产品研发。E-mail:
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The Food Industry, 2023, 44(1): 100-105., articleTitle=Optimization of the extraction process of Proanthocyanidins from grape seeds, refAbstract=null), Reference(id=1167030739922072195, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, doi=null, pmid=null, pmcid=null, year=2013, volume=29, issue=6, pageStart=86, pageEnd=87, url=null, language=null, rfNumber=[30], rfOrder=52, authorNames=乔振杰, 李晓艳, journalName=德州学院学报, refType=null, unstructuredReference=乔振杰, 李晓艳. 野生药用植物萹蓄总黄酮的提取工艺研究[J]. 德州学院学报, 2013, 29(6): 86-87, 94., articleTitle=野生药用植物萹蓄总黄酮的提取工艺研究, refAbstract=null), Reference(id=1167030740014346884, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, doi=null, pmid=null, pmcid=null, year=2013, volume=29, issue=6, pageStart=86, pageEnd=87, url=null, language=null, rfNumber=[30], rfOrder=53, authorNames=QIAO ZJ, LI XY, journalName=Journal of Dezhou University, refType=null, unstructuredReference=QIAO ZJ, LI XY. The research on the exacting of total Flavanoid in wild medicinal plants of Polygonum aviculare[J]. Journal of Dezhou University, 2013, 29(6): 86-87, 94., articleTitle=The research on the exacting of total Flavanoid in wild medicinal plants of Polygonum aviculare, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1167030733202797081, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, xref=null, ext=[AuthorCompanyExt(id=1167030733211185690, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, companyId=1167030733202797081, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Weinan Inspection and Testing Research Institute, Weinan 714000, China, 2. 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Factor and level tables of response surface analysis

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因素
X1(超声时间)
/min
X2(乙醇体积
分数)/%
X3(料液比)
(g:mL)
-1 20 50 1:30
0 30 60 1:40
1 40 70 1:50
), ArticleFig(id=1167030736075895367, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, language=CN, label=表1, caption=

响应面分析因子和水平表

, figureFileSmall=null, figureFileBig=null, tableContent=
因素
X1(超声时间)
/min
X2(乙醇体积
分数)/%
X3(料液比)
(g:mL)
-1 20 50 1:30
0 30 60 1:40
1 40 70 1:50
), ArticleFig(id=1167030736126227016, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, language=EN, label=Table 2, caption=

Results of response surface analysis

, figureFileSmall=null, figureFileBig=null, tableContent=
试验号 因素 Y(原花青素
提取量)/%
X1(超声时间)
/min
X2(乙醇
体积分数)/%
X3(料液比)
(g:mL)
1 30 60 40 2.34
2 30 50 30 2.06
3 30 50 50 2.02
4 30 60 40 2.35
5 30 60 40 2.34
6 40 60 30 2.02
7 40 50 40 1.97
8 30 70 50 1.99
9 20 60 50 1.77
10 30 60 40 2.36
11 20 70 40 1.88
12 30 60 40 2.23
13 40 60 50 1.83
14 40 70 40 1.75
15 30 70 30 1.80
16 20 60 30 1.85
17 20 50 40 1.93
), ArticleFig(id=1167030736210113097, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, language=CN, label=表2, caption=

响应面分析结果

, figureFileSmall=null, figureFileBig=null, tableContent=
试验号 因素 Y(原花青素
提取量)/%
X1(超声时间)
/min
X2(乙醇
体积分数)/%
X3(料液比)
(g:mL)
1 30 60 40 2.34
2 30 50 30 2.06
3 30 50 50 2.02
4 30 60 40 2.35
5 30 60 40 2.34
6 40 60 30 2.02
7 40 50 40 1.97
8 30 70 50 1.99
9 20 60 50 1.77
10 30 60 40 2.36
11 20 70 40 1.88
12 30 60 40 2.23
13 40 60 50 1.83
14 40 70 40 1.75
15 30 70 30 1.80
16 20 60 30 1.85
17 20 50 40 1.93
), ArticleFig(id=1167030736277221962, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, language=EN, label=Table 3, caption=

Variance analysis of regression model

, figureFileSmall=null, figureFileBig=null, tableContent=
方差
来源
平方和 自由度 均方 F P
模型 0.710000 9 0.079000 11.97 0.0018**
X1 0.003062 1 0.003062 0.46 0.5177
X2 0.039000 1 0.039000 5.84 0.0464*
X3 0.002021 1 0.002021 0.31 0.5972
X1X2 0.007453 1 0.007453 1.13 0.3232
X1X3 0.002836 1 0.002836 0.43 0.5331
X2X3 0.013000 1 0.013000 2.02 0.1986
X12 0.310000 1 0.310000 46.33 0.0003**
X22 0.130000 1 0.130000 18.94 0.0033**
X32 0.150000 1 0.150000 22.54 0.0021**
残差 0.046000 7 0.006600
失拟项 0.035000 3 0.012000 4.29 0.0967
纯误差 0.011000 4 0.002739
总回归 0.760000 16
R2 R2=0.9389, R2Adj=0.8605
), ArticleFig(id=1167030736344330827, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437191147778672, language=CN, label=表3, caption=

回归模型方差分析

, figureFileSmall=null, figureFileBig=null, tableContent=
方差
来源
平方和 自由度 均方 F P
模型 0.710000 9 0.079000 11.97 0.0018**
X1 0.003062 1 0.003062 0.46 0.5177
X2 0.039000 1 0.039000 5.84 0.0464*
X3 0.002021 1 0.002021 0.31 0.5972
X1X2 0.007453 1 0.007453 1.13 0.3232
X1X3 0.002836 1 0.002836 0.43 0.5331
X2X3 0.013000 1 0.013000 2.02 0.1986
X12 0.310000 1 0.310000 46.33 0.0003**
X22 0.130000 1 0.130000 18.94 0.0033**
X32 0.150000 1 0.150000 22.54 0.0021**
残差 0.046000 7 0.006600
失拟项 0.035000 3 0.012000 4.29 0.0967
纯误差 0.011000 4 0.002739
总回归 0.760000 16
R2 R2=0.9389, R2Adj=0.8605
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响应面法优化超声提取萹蓄原花青素工艺
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尚琪 * , 宋玉 , 刘宇 , 王建平 , 代明俊
食品安全质量检测学报 | 食品加工与工艺 2025,16(11): 307-313
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食品安全质量检测学报 | 食品加工与工艺 2025, 16(11): 307-313
响应面法优化超声提取萹蓄原花青素工艺
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尚琪* , 宋玉, 刘宇, 王建平, 代明俊
作者信息
  • 渭南市检验检测研究院, 渭南 714000, 2. 贵州万胜药业有限责任公司, 遵义 563000

通讯作者:

* 尚琪(1990—), 男, 硕士, 工程师, 主要研究方向为食品及相关产品的检验检测, 药食同源植物天然产物的提取与分离及产品研发。E-mail:
Optimization of ultrasonic extraction process of proanthocyanidins from Polygonum aviculare by response surface method
Qi SHANG* , Yu SONG, Yu LIU, Jian-Ping WANG, Ming-Jun DAI
Affiliations
  • Weinan Inspection and Testing Research Institute, Weinan 714000, China, 2. Guizhou Wansheng Pharmaceutical Co., Ltd., Zunyi 563000, China
出版时间: 2025-06-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250228007
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目的 通过利用响应面法优化超声辅助提取食药植物萹蓄中原花青素的工艺, 获取最佳工艺条件。方法 通过单因素试验考察影响提取量的主要因素, 在单因素试验结果的基础上, 用Box-Behnken设计及响应面分析法, 对萹蓄中原花青素提取量的影响单因素乙醇体积分数、超声时间、料液比进行优化, 获取最佳提取工艺条件。结果 萹蓄原花青素超声辅助提取最佳工艺条件优化结果分别为: 超声时间34 min、乙醇体积分数65%、料液比1:46 (g:mL), 在此工艺条件下, 原花青素的最终提取量达到2.18%, 与模型验证值2.12%相符合。结论 本研究表明响应面法优化得出的工艺条件参数是有效可行的, 成功地应用响应面法优化了超声辅助提取萹蓄中原花青素的提取参数, 为萹蓄原花青素的后期推广应用提供研究基础。

响应面法  /  超声提取  /  萹蓄  /  原花青素

Objective To optimize the ultrasonic-assisted extraction process of proanthocyanidins from Polygonum aviculare by response surface method, to obtain the best extraction conditions. Methods The main factors affecting the extraction amount were investigated through single-factor experiments. Based on the results of the single-factor experiments, the Box-Behnken design and response surface methodology were used to optimize the single factors, including the volume fraction of ethanol, ultrasonic time, and solid-liquid ratio, which had an impact on the extraction amount of proanthocyanidins from Polygonum aviculare. The optimal extraction process conditions were obtained. Results The optimized results of the optimal process conditions for ultrasonic-assisted extraction of proanthocyanidins from Polygonum aviculare were as follows: The ultrasonic time was 34 min, the volume fraction of ethanol was 65%, and the solid-liquid ratio was 1:46 (g:mL). Under these process conditions, the final extraction amount of proanthocyanidins reached 2.18%, which was consistent with the model validation value of 2.12%. Conclusion This study shows that the process condition parameters optimized by the response surface methodology are effective and feasible. The response surface methodology is successfully applied to optimize the extraction parameters of ultrasonic-assisted extraction of proanthocyanidins from Polygonum aviculare, providing a research basis for the later promotion and application of proanthocyanidins from Polygonum aviculare.

response surface method  /  ultrasonic extraction  /  Polygonum aviculare  /  proanthocyanidins
尚琪, 宋玉, 刘宇, 王建平, 代明俊. 响应面法优化超声提取萹蓄原花青素工艺. 食品安全质量检测学报, 2025 , 16 (11) : 307 -313 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250228007
Qi SHANG, Yu SONG, Yu LIU, Jian-Ping WANG, Ming-Jun DAI. Optimization of ultrasonic extraction process of proanthocyanidins from Polygonum aviculare by response surface method[J]. Journal of Food Safety & Quality, 2025 , 16 (11) : 307 -313 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250228007
萹蓄(Polygonum aviculare)为蓼科(Polygonaceae)蓼属一年生或多年生草本植物, 又名地萹蓄、萹竹、道生草等, 山坡、田野、路旁等均可生处, 国内均有分布[1]。萹蓄为药食两用植物, 全草可以药用, 具有利尿, 杀虫、止痒功效, 主治热淋涩痛、小便短赤、湿疹、阴痒等病[2-5]。萹蓄含有多种化学成分, 主要含有阿魏酸、没食子酸、儿茶素等黄酮类化合物, 同时含有丰富的水溶性多糖和游离氨基酸[6-8]; 现代药理研究也证实其利尿、降压、保肝、抗氧化、抗肥胖等的生理活性[8-11]
原花青素是聚多酚类物质, 以不同数量的表儿茶素、没食子酸或儿茶素以C4-C6(或C4-C8)键缩合而成, 广泛存在于各种植物的不同部位中[12-13]。不少研究证明原花青素具有抗氧化、抗高血糖、保护心血管和抗辐射的功效[14], 主要应用于食品、特殊食品和化妆品领域[15-17]
目前对于萹蓄的研究, 主要集中在化学成分分析及药理活性的研究[11,18-20], 也有报道研究了萹蓄中总黄酮提取工艺的研究[21], 而其中原花青素的提取工艺的研究报道较少。超声波辅助提取技术是一项提取天然产物应用较广且环保的技术, 具有提率高、时间短、成本低、条件温和且保证产物质量高等优点[22-24]。基于此, 本研究采用超声提取技术, 响应面分析法优化提取萹蓄中的原花青素工艺参数, 以期为萹蓄中原花青素的开发利用提供后续科研的依据。
萹蓄: 2024年8月采自定西市渭源县会川镇, 为一年生或多年生的蓼科(Polygonaceae)蓼属(Polygonum)植物萹蓄(Polygonum aviculare)的地上部分。原料淋洗干净, 自然阴干, 粉碎, 过60目筛, 保存备用。
(+)-儿茶素(纯度≥98%, 上海金穗生物科技有限公司); 甲醇、丙酮、无水乙醇、浓盐酸、香草醛(分析纯, 国药集团化学试剂有限公司)。
DL-820E智能超声波清洗器(上海之信仪器有限公司); SHZ-2000型双配套循环水式多用真空泵(河南省巩义市英峪予华仪器厂); T9CS双光束紫外可见光分光光度计(北京普析通用仪器有限责任公司); HH-4数显恒温水浴锅(国华电器有限公司); Sarstorius BT125D电子天平(精度0.1 mg, 德国赛多利斯科学仪器有限公司)。
采用香草醛-盐酸法[25]并加以修改。标准曲线的制备: 分别吸取质量浓度0.2 mg/mL的儿茶素0.1、0.2、0.3、0.4、0.5 mL, 无水乙醇补至0.5 mL。加入3 mL浓度为4%香草醛和浓盐酸1.5 mL, 加塞混匀, 室温下避光反应15 min, 于500 nm处测其吸光值, 以儿茶素质量为横坐标(X, mg), 吸光度为纵坐标(Y), 制作标准曲线。
原花青素含量的测定: 准确吸取提取液0.2 mL, 提取剂补至0.5 mL。根据标准曲线的制作方法, 测定吸光度A, 按公式(1)计算原花青素的提取量:
原花青素的提取量/%=$\frac{V\times C}{1000\times W}$×100%
式中: V为试样定容体积, mL; C为试样中原花青素质量浓度, mg/mL; W为试样质量, g; 1000为单位换算系数。
采用体积分数乙醇(70%)、甲醇、丙酮溶剂法提取原花青素, 比对提取剂对萹蓄原花青素的提取效果。
(1)超声时间对原花青素提取效果的影响
称取萹蓄粉末1 g, 分别至5个锥形瓶中。按料液比1:20 (g:mL)加入体积分数为70%乙醇, 40 ℃下, 超声频率560 W的条件下, 超声10、20、30、40、50 min, 冷却后减压过滤, 收集滤液定容至50 mL, 根据1.3.1节方法测定提取液中的原花青素的提取量, 比对超声时间对提取效果的影响。
(2)乙醇体积分数对原花青素提取效果的影响
称取萹蓄粉末1 g, 分别至5个锥形瓶中, 40 ℃下, 超声频率560 W的条件下, 按料液比1:40 (g:mL)分别加入40%、50%、60%、70%、80%乙醇(体积分数), 超声30 min, 冷却后减压过滤, 收集滤液定容至50 mL, 根据1.3.1节方法测定提取液中的原花青素的提取量, 比较乙醇体积分数对提取效果的影响。
(3)超声温度对原花青素提取效果的影响
称取萹蓄粉末1 g, 分别至5个锥形瓶中。按料液比1:20 (g:mL)加入60%的乙醇, 在温度30、40、50、60、70 ℃下560 W超声频率提取30 min, 冷却后减压过滤, 收集滤液定容至50 mL, 根据1.3.1节介绍的方法测定提取液中的原花青素的提取量, 比较超声温度对提取效果的影响。
(4)料液比对原花青素提取效果的影响
称取萹蓄粉末1 g, 分别至5个锥形瓶中。按料液比1:10、1:20、1:30、1:40、1:50 (g:mL)加入60%的乙醇, 在40 ℃下、超声频率380 W、超声提取30 min, 冷却后减压过滤, 除1:50 (g:mL)的滤液需要减压浓缩后定容, 其他滤液直接定容至50 mL, 根据1.3.1节介绍的方法测定提取液中的原花青素的提取量, 比较料液比对提取效果的影响。
(5)超声功率对原花青素提取效果的影响
称取萹蓄粉末1 g, 分别至5个锥形瓶中。按料液比为1:40 (g:mL)加入60%乙醇, 分别在超声功率380、440、500、560、620 W, 温度40 ℃的条件下, 超声提取30 min, 冷却后减压过滤, 收集滤液定容至50 mL, 根据1.3.1节介绍的方法测定提取液中的原花青素的提取量, 比较超声功率对提取效果的影响。
根据单因素试验结果, 依据Box-Behnken 试验设计原理, 选(超声时间、乙醇体积分数、料液比)3个影响较大的因素进行响应面设计, 优化萹蓄原花青素提取工艺。设计方案见表1
采用Excel 2003软件对试验数据进行处理, 多重比较采用SPSS 16软件进行Duncan新复极差法分析。响应面试验数据利用Desingn Expert 8.0软件进行多元回归和方差分析, 确定模型和因素的显著性。
依据标准曲线进行线性回归, 得线性回归方程Y=5.1643X+0.008, r2=0.9995, 按回归方程计算各提取剂对原花青素提取效果的影响, 结果如图1所示。从图1可以看出, 提取原花青素的效果为乙醇>甲醇>丙酮。多重比较显示, 体积分数70%乙醇与甲醇、丙酮所提取的原花青素相比较差异显著(P<0.05); 因为原花青素属于多酚类化合物, 含有大量的羟基, 具有一定的极性[25], 易溶于乙醇和甲醇中, 乙醇毒性小, 提取效率高。因此, 选乙醇作为提取溶剂。
图2a可知, 原花青素提取量在10~30 min中呈逐增趋势, 数据显示, 提取时间对原花青素提取量的影响相比较差异显著(P<0.05); 当提取时间在30 min以上时, 随时间加长提取量反呈下降趋势。由于超声提取时间过长, 导致原花青素部分酚结构被破坏[26]; 当提取时间过短时, 原花青素未能从萹蓄细胞中溶出。因此时间过长过短造成原花青素的提取量降低。最佳提取时间确定为30 min。
图2b可知, 原花青素提取量在乙醇体积分数40%~60%之间逐渐增加, 超过60%时, 提取量呈缓慢下降趋势, 分析数据, 乙醇体积分数对原花青素提取量的影响相比较差异显著(P<0.05)。可能是随着乙醇体积分数增大, 溶出物太杂多, 与成分较为复杂有关[27], 乙醇浓度过大或过小会使提取量降低, 因此确定提取的适宜乙醇体积分数为60%。
图2c可以看出, 随着超声温度的增加, 提取量也在逐渐升高, 但变化比较平缓, 温度70 ℃时与30 ℃和40 ℃相比较分别增加了0.25%和0.19%, 分析原因可能是, 70 ℃温度范围内, 温度高分子运动加快, 提取量增加。超声与加热联合处理, 可以引起细胞局部微裂缝的形成, 导致细胞原花青素释放, 使其提取量增加[28], 但超声温度过高会使更多的杂质析出, 造成续纯化工艺复杂。因此, 从节能考虑, 在响应面优化中固定温度为40 ℃。
图2d在提取料液比1:10~1:40 (g:mL)之间原花青素提取量逐渐增加, 当料液比超过1:40 (g:mL)时, 随料液比增加提取量缓慢下降。当达到1:40 (g:mL)时有效成分已基本提取完毕, 再增加料液比会使更多的杂质析出, 导致原花青素提取量降低, 分析数据, 料液比对原花青素提取量的影响相比较差异显著(P<0.05)。因此料液比过大或过小都会使提取量降低。故确定最佳提取料液比为1:40 (g:mL)。
图2e可以看出, 随着超声功率的逐渐增加, 提取量逐渐增大, 在功率500 W增加较大, 相对于380 W原花青素增加了0.034%。随后又逐渐下降, 但变化幅度很小, 分析数据, 超声功率对原花青素提取量的影响相比较差异不显著(P>0.05)。原因是超声波的破碎作用不仅取决于超声波的功率, 还取决于被破碎体系的性质, 超声波对萹蓄细胞的破碎程度在380 W时原花青素已基本溶出, 继续加大功率不再对原花青素提取产生影响。节能考虑, 超声功率以380 W为宜。由于随功率的变化原花青素变化幅度不明显, 在响应面优化中固定功率为380 W。
响应变量为超声时间(X1)、乙醇体积分数(X2)和料液比(X3), 原花青素提取量(Y)为响应值, 采用Design-Expert 8.0软件的Box-Behnken设计, 进行试验, 试验结果为表2。进行多项式回归拟合表2数据, 建立响应面法优化超声提取萹蓄原花青素的回归方程:
Y=2.32450+0.019564X1-0.069392X2-0.015895X3-0.043164X1X2-0.026626X1X3+0.057675X2X3-0.269478X12-0.172309X22-0.187976X32
表2结果进行方差分析, 结果见表3。二次响应面回归模型F为11.97, 并且P<0.01, 说明该方程模型极显著, 可预测不同超声条件下原花青素的提取量。方程式中, 一次项X2P<0.05, 与响应值回归关系较为显著, 说明乙醇体积分数对原花青素提取量影响显著。二次项X12X22X32P<0.01, 与响应值回归关系极显著, 说明对原花青素提取量影响极显著。其交互项X1X2X2X3X1X3均不显著。该模型失拟项P为0.0967大于0.05, 模型失拟不显著, 说明该模型有效, 预测决定系数R2=0.9389, 调整系数R2Adj=0.8605, 试验结果有86.05%受试验因素的影响。因此, 试验结果真实可靠, 运用该模型可以较好对原花青素的提取量进行分析和预测。从响应面回归方程一次项F检验值大小可以得出, 影响萹蓄原花青素提取量大小的因素依次为乙醇体积分数(X2)>超声时间(X1)>料液比(X3)。
响应面可以直接反映出各因子对响应值的影响大小, 由等高线图可以直接看出最优条件下各因子的取值。从图3a~d可以看出, 在超声时间轴向上较乙醇体积分数、料液比轴向等高线较密集, 说明对响应值的影响效果比较大, 影响效果依次为乙醇体积分数>超声时间>料液比[29], 但是从等高线图的形状可以看出, 超声时间与乙醇体积分数和料液比的等高线为圆形, 表示两因素交互作用不显著, 这和方差分析的结果相一致。由图3e~f乙醇体积分数和料液比的响应面及等高线图可以看出, 乙醇体积分数对原花青素提取量的影响最明显, 表现为曲线相对较陡, 等高线在乙醇体积分数轴向上密度较大, 且为椭圆形, 因此说明两交互作用较为明显。
利用Design-Expert 8.0软件对工艺参数优化, 得出从萹蓄中提取原花青素的工艺条件为: 乙醇体积分数65.35%, 液料比1:46.12 (g:mL), 超声时间34.4 min, 理论值可达2.12%。为检验该方法的可靠性, 考虑实际操作的便利, 将最佳工艺参数修正为: 乙醇体积分数65%, 液料比1:46 (g:mL), 超声时间34 min, 经5次平行试验, 得实际平均提取量为2.18%, 变异系数是2.22%, 试验结果与模型预测值2.12%符合良好。结果表明, 该模型较好地反映出萹蓄中原花青素提取的条件, 证明了响应面法优化提取条件参数的可行性。
采用传统乙醇回流提取法, 按料液比1:40 (g:mL), 乙醇体积分数65%,, 温度80 ℃, 浸提3 h条件下, 提取量为1.91%, 而超声波辅助提取优化工艺的提取量达2.18%, 与传统乙醇浸提法比对, 提取量提高了14%。提取时间缩短了2.5 h, 提取温度降低了40 ℃。提取量显著提高, 有效缩短了提取时间, 提取过程高效节能。
本研究以萹蓄为研究材料, 比较几种溶剂对原花青素提取的效果, 结果表明, 乙醇提取效果较好, 因为原花青素易溶于水, 醇类等有机溶剂, 乙醇提取毒性小, 可以避免多糖、蛋白质和果胶等成分对目标产物提取的影响[30]。因此以乙醇为提取剂, 在单因素试验的基础上, 通过Box-Behnken试验设计及响应面分析, 对超声提取萹蓄原花青素的工艺进行了优化, 建立了二次多项式回归方程数学模型, 确定了目标函数与工艺条件之间的关系。各因素对提取工艺影响的主次顺序为乙醇体积分数、超声时间、料液比。利用模型得最优工艺参数: 乙醇体积分数65%、超声时间34 min、液料比1:46 (g:mL)。在此条件下, 萹蓄原花青素的提取量可达2.18%, 与理论预测值相符, 说明本研究的提取是有效、可行的, 后续将对其纯化、鉴定, 并跟踪检测其活性, 为萹蓄活性成分的开发利用奠定基础。
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250228007
  • 接收时间:2025-02-28
  • 首发时间:2025-07-14
  • 出版时间:2025-06-15
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    渭南市检验检测研究院, 渭南 714000, 2. 贵州万胜药业有限责任公司, 遵义 563000

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* 尚琪(1990—), 男, 硕士, 工程师, 主要研究方向为食品及相关产品的检验检测, 药食同源植物天然产物的提取与分离及产品研发。E-mail:
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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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