Article(id=1153433697442320889, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433686872679135, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241030009, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1730217600000, receivedDateStr=2024-10-30, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752929623232, onlineDateStr=2025-07-19, pubDate=1744646400000, pubDateStr=2025-04-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752929623232, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752929623232, creator=13701087609, updateTime=1752929623232, updator=13701087609, issue=Issue{id=1153433686872679135, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='7', pageStart='1', pageEnd='322', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752929620712, creator=13701087609, updateTime=1757656380159, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1173259152974561742, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433686872679135, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1173259152978756047, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153433686872679135, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=94, endPage=101, ext={EN=ArticleExt(id=1153433698125992461, articleId=1153433697442320889, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Study on the change law and influencing factors of biogenic amines during soy sauce fermentation, columnId=1151895322692776479, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Analysis and Monitoring of Toxic and Harmful Substances in Food, runingTitle=null, highlight=null, articleAbstract=

Objective To study the change law of biogenic amines in the process of soy sauce fermentation and explore the influence of production conditions on the accumulation of biogenic amines. Methods The content of biological amines in soy sauce was analyzed at different fermentation times by high performance liquid chromatography (HPLC) technology. Meanwhile, the effects of different saltwater concentrations (15%, 20%, 25%), fermentation temperatures (20, 30, 40 ℃), and yeast additions (0.05%, 0.10%, 0.15%) on the production of biological amines in soy sauce fermentation were explored. Results The content of biological amines in soy sauce presented an increasing trend first and then decreasing as the fermentation time was prolonged; the lowest content of biological amines in soy sauce after fermentation ends was 368.9 mg/L at a temperature of 40 ℃; the lowest content of biological amines in soy sauce after fermentation ends was 376.41 mg/L at a saltwater concentration of 25%; the lowest content of biological amines in soy sauce after fermentation ends was 403.45 mg/L when the yeast addition was 0.05%. Conclusion Biogenic amines are harmful substances in soy sauce fermentation process, and their excessive content can have certain impacts on human health. By adjusting the saltwater concentration, fermentation temperature, and yeast addition, it is possible to effectively inhibit the formation of biogenic amines during soy sauce fermentation, thereby effectively improving the safety of soy sauce.

, correspAuthors=Qi-Xu FU, 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=Hui WANG, Xue-Mei SHI, Bing-Yu HUANG, Lin WANG, Zhen YANG, Li-Na PAN, Xian-Bin WANG, Qi-Xu FU, Dan LI), CN=ArticleExt(id=1153433727842636314, articleId=1153433697442320889, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=酱油发酵过程中生物胺的变化规律及影响因素研究, columnId=1151895322898297380, journalTitle=食品安全质量检测学报, columnName=本期专题:食品中有毒有害物质分析与监测, runingTitle=null, highlight=null, articleAbstract=

目的 研究酱油发酵过程中生物胺的变化规律, 并探究生产条件对生物胺积累的影响。方法 利用高效液相色谱(high performance liquid chromatography, HPLC)技术分析了酱油不同发酵时间下生物胺的含量; 同时探究了不同盐水浓度(15%、20%、25%)、发酵温度(20、30、40 ℃)、酵母添加量(0.05%、0.10%、0.15%)对酱油发酵过程中生物胺产生的影响。结果 酱油中生物胺含量随着发酵时间的延长呈现先增加后降低的趋势; 在40 ℃的温度下, 酱油发酵结束后的生物胺含量最低, 为368.9 mg/L; 在盐水浓度为25%时, 酱油发酵结束后的生物胺含量最低, 为376.41 mg/L; 酵母添加量为0.05%时, 酱油发酵结束后的生物胺含量最低, 为403.45 mg/L。结论 生物胺作为酱油发酵过程中的有害物质, 其含量过高会对人体健康产生一定影响, 通过改变盐水浓度、发酵温度和酵母添加量, 可以有效抑制酱油发酵过程中生物胺的形成, 从而有效提高酱油的安全性。

, correspAuthors=傅其旭, authorNote=null, correspAuthorsNote=
* 傅其旭(1985—), 男, 工程师, 主要研究方向为酒类及调味品酿造。E-mail:
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王惠(1998—), 女, 硕士, 主要研究方向为食品安全检测。E-mail:

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王惠(1998—), 女, 硕士, 主要研究方向为食品安全检测。E-mail:

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王惠(1998—), 女, 硕士, 主要研究方向为食品安全检测。E-mail:

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注: a. 酱油发酵过程中总酸的变化情况; b. 酱油发酵过程中氨基酸态氮的变化情况; c. 酱油发酵过程中全氮的变化情况。

, figureFileSmall=NhFl0U+agKXBpVQ7ZjCQ2Q==, figureFileBig=2hCnb0ZtSK4cmkfYhY55jg==, tableContent=null), ArticleFig(id=1173278649794048766, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Fig.3, caption=Derivative chromatogram of biogenic amines in soy sauce, figureFileSmall=fmNGDurds/Hf1Fmlk2arKQ==, figureFileBig=DIZVruvqDTMtNkC6MNfIDA==, tableContent=null), ArticleFig(id=1173278649869546243, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=图3, caption=酱油中生物胺的衍生色谱图, figureFileSmall=fmNGDurds/Hf1Fmlk2arKQ==, figureFileBig=DIZVruvqDTMtNkC6MNfIDA==, tableContent=null), ArticleFig(id=1173278649945043719, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Fig.4, caption=Changes of total biogenic amines during soy sauce fermentation (a) and composition analysis of total biogenic amines in soy sauce fermentation process (b), figureFileSmall=pfLTlVnkMiza8+LhQKLtAQ==, figureFileBig=VCpUdPSk53zB1qZ377MmiQ==, tableContent=null), ArticleFig(id=1173278650070872842, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=图4, caption=酱油发酵过程中总生物胺的变化规律(a)和酱油发酵过程中总生物胺的组成分析(b), figureFileSmall=pfLTlVnkMiza8+LhQKLtAQ==, figureFileBig=VCpUdPSk53zB1qZ377MmiQ==, tableContent=null), ArticleFig(id=1173278650171536140, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Fig.5, caption=Effects of brine concentration on biogenic amine production during soy sauce fermentation (a) and composition analysis of total biogenic amines under different saline concentrations (b), figureFileSmall=bCtrKDOZS4jtNJWUuFruzA==, figureFileBig=xhj1OmrUTcg/VQbmHzWx+g==, tableContent=null), ArticleFig(id=1173278650268005135, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=图5, caption=盐水浓度对酱油发酵过程中生物胺产生的影响(a)和在不同盐水浓度下总生物胺的组成分析(b), figureFileSmall=bCtrKDOZS4jtNJWUuFruzA==, figureFileBig=xhj1OmrUTcg/VQbmHzWx+g==, tableContent=null), ArticleFig(id=1173278650389639953, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Fig.6, caption=Effects of fermentation temperature on biogenic amines production during soy sauce fermentation (a) and composition analysis of total biogenic amines at different fermentation temperatures (b), figureFileSmall=jB8tigut/sxjoHqDA/NjAQ==, figureFileBig=WZIcri/w3N9mkwwaaHDfEQ==, tableContent=null), ArticleFig(id=1173278650511274772, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=图6, caption=发酵温度对酱油发酵过程中生物胺产生的影响(a)和在不同发酵温度下总生物胺的组成分析(b), figureFileSmall=jB8tigut/sxjoHqDA/NjAQ==, figureFileBig=WZIcri/w3N9mkwwaaHDfEQ==, tableContent=null), ArticleFig(id=1173278650632909591, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Fig.7, caption=Effects of yeast addition on biogenic amine production during soy sauce fermentation (a) and composition analysis of total biogenic amines under different yeast addition (b), figureFileSmall=h5JNTSKSEVXI4ZhgboeYbg==, figureFileBig=7hM3uX4lrXu9qV/v4rQgmA==, tableContent=null), ArticleFig(id=1173278650758738714, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=图7, caption=酵母添加量对酱油发酵过程中生物胺产生的影响(a)和在不同酵母添加量下总生物胺的组成分析(b), figureFileSmall=h5JNTSKSEVXI4ZhgboeYbg==, figureFileBig=7hM3uX4lrXu9qV/v4rQgmA==, tableContent=null), ArticleFig(id=1173278650813264669, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=EN, label=Table 1, caption=

Biogenic amines and their toxic effects

, figureFileSmall=null, figureFileBig=null, tableContent=
生物胺类别 生物胺 毒性作用
芳香胺 酪胺 引起偏头痛、神经系统疾病、呕吐和高血压等[19]
β-苯乙胺 消除神经系统中去甲肾上腺素, 引起高血压和偏头痛[20]
脂肪胺 精胺 参与致癌、肿瘤侵袭和转移[21]
腐胺 引起心跳加速、高血压, 具有致癌作用[19]
尸胺 与亚硝酸盐发生反应, 形成致癌的亚硝基胺[19]
杂环胺 亚精胺 与亚硝酸盐发生反应, 形成致癌的亚硝基胺[19]
色胺 引起哮喘、血压升高和消化系统失调[19]
组胺 引起头痛、呼吸困难、支气管痉挛、皮疹、水肿和腹泻[22]
), ArticleFig(id=1173278650884567842, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153433697442320889, language=CN, label=表1, caption=

生物胺类别及其毒性作用

, figureFileSmall=null, figureFileBig=null, tableContent=
生物胺类别 生物胺 毒性作用
芳香胺 酪胺 引起偏头痛、神经系统疾病、呕吐和高血压等[19]
β-苯乙胺 消除神经系统中去甲肾上腺素, 引起高血压和偏头痛[20]
脂肪胺 精胺 参与致癌、肿瘤侵袭和转移[21]
腐胺 引起心跳加速、高血压, 具有致癌作用[19]
尸胺 与亚硝酸盐发生反应, 形成致癌的亚硝基胺[19]
杂环胺 亚精胺 与亚硝酸盐发生反应, 形成致癌的亚硝基胺[19]
色胺 引起哮喘、血压升高和消化系统失调[19]
组胺 引起头痛、呼吸困难、支气管痉挛、皮疹、水肿和腹泻[22]
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酱油发酵过程中生物胺的变化规律及影响因素研究
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王惠 1 , 石学梅 1 , 黄冰羽 1 , 王淋 1 , 杨镇 1 , 潘丽娜 1 , 王宪斌 1 , 傅其旭 1, * , 李丹 2
食品安全质量检测学报 | 本期专题:食品中有毒有害物质分析与监测 2025,16(7): 94-101
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食品安全质量检测学报 | 本期专题:食品中有毒有害物质分析与监测 2025, 16(7): 94-101
酱油发酵过程中生物胺的变化规律及影响因素研究
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王惠1 , 石学梅1, 黄冰羽1, 王淋1, 杨镇1, 潘丽娜1, 王宪斌1, 傅其旭1, * , 李丹2
作者信息
  • 1.四川元景达食品有限公司, 泸州 646000
  • 2.泸州老窖集团有限责任公司, 泸州 646000
  • 王惠(1998—), 女, 硕士, 主要研究方向为食品安全检测。E-mail:

通讯作者:

* 傅其旭(1985—), 男, 工程师, 主要研究方向为酒类及调味品酿造。E-mail:
Study on the change law and influencing factors of biogenic amines during soy sauce fermentation
Hui WANG1 , Xue-Mei SHI1, Bing-Yu HUANG1, Lin WANG1, Zhen YANG1, Li-Na PAN1, Xian-Bin WANG1, Qi-Xu FU1, * , Dan LI2
Affiliations
  • 1. Sichuan Yuanjingda Food Co., Ltd., Luzhou 646000, China
  • 2. Luzhou Laojiao Group Co., Ltd., Luzhou 646000, China
出版时间: 2025-04-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241030009
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目的 研究酱油发酵过程中生物胺的变化规律, 并探究生产条件对生物胺积累的影响。方法 利用高效液相色谱(high performance liquid chromatography, HPLC)技术分析了酱油不同发酵时间下生物胺的含量; 同时探究了不同盐水浓度(15%、20%、25%)、发酵温度(20、30、40 ℃)、酵母添加量(0.05%、0.10%、0.15%)对酱油发酵过程中生物胺产生的影响。结果 酱油中生物胺含量随着发酵时间的延长呈现先增加后降低的趋势; 在40 ℃的温度下, 酱油发酵结束后的生物胺含量最低, 为368.9 mg/L; 在盐水浓度为25%时, 酱油发酵结束后的生物胺含量最低, 为376.41 mg/L; 酵母添加量为0.05%时, 酱油发酵结束后的生物胺含量最低, 为403.45 mg/L。结论 生物胺作为酱油发酵过程中的有害物质, 其含量过高会对人体健康产生一定影响, 通过改变盐水浓度、发酵温度和酵母添加量, 可以有效抑制酱油发酵过程中生物胺的形成, 从而有效提高酱油的安全性。

酱油  /  发酵  /  生产条件  /  生物胺  /  变化规律

Objective To study the change law of biogenic amines in the process of soy sauce fermentation and explore the influence of production conditions on the accumulation of biogenic amines. Methods The content of biological amines in soy sauce was analyzed at different fermentation times by high performance liquid chromatography (HPLC) technology. Meanwhile, the effects of different saltwater concentrations (15%, 20%, 25%), fermentation temperatures (20, 30, 40 ℃), and yeast additions (0.05%, 0.10%, 0.15%) on the production of biological amines in soy sauce fermentation were explored. Results The content of biological amines in soy sauce presented an increasing trend first and then decreasing as the fermentation time was prolonged; the lowest content of biological amines in soy sauce after fermentation ends was 368.9 mg/L at a temperature of 40 ℃; the lowest content of biological amines in soy sauce after fermentation ends was 376.41 mg/L at a saltwater concentration of 25%; the lowest content of biological amines in soy sauce after fermentation ends was 403.45 mg/L when the yeast addition was 0.05%. Conclusion Biogenic amines are harmful substances in soy sauce fermentation process, and their excessive content can have certain impacts on human health. By adjusting the saltwater concentration, fermentation temperature, and yeast addition, it is possible to effectively inhibit the formation of biogenic amines during soy sauce fermentation, thereby effectively improving the safety of soy sauce.

soy sauce  /  fermentation  /  production conditions  /  biogenic amine  /  change law
王惠, 石学梅, 黄冰羽, 王淋, 杨镇, 潘丽娜, 王宪斌, 傅其旭, 李丹. 酱油发酵过程中生物胺的变化规律及影响因素研究. 食品安全质量检测学报, 2025 , 16 (7) : 94 -101 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241030009
Hui WANG, Xue-Mei SHI, Bing-Yu HUANG, Lin WANG, Zhen YANG, Li-Na PAN, Xian-Bin WANG, Qi-Xu FU, Dan LI. Study on the change law and influencing factors of biogenic amines during soy sauce fermentation[J]. Journal of Food Safety & Quality, 2025 , 16 (7) : 94 -101 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241030009
酱油作为我国传统调味品的一种, 在家庭烹饪、餐饮渠道、食品加工等领域发挥着重要作用[1-3]。在酱油发酵过程中, 蛋白质被米曲霉产生的蛋白酶水解成小分子肽和游离氨基酸, 成为酱油鲜味物质的主要来源[4-6]。在此过程中, 其余酶类也会参与反应, 例如氨基酸脱羧酶会导致生物胺的生成[7-9]
生物胺主要是一类低分子量的含氮有机化合物, 具有生物毒性[10-12]。食品中生物胺的产生可分为内源性以及外源性。内源性指原料自身醛酮类化合物的氨基化和转氨基作用产生的生物胺; 外源性主要是原料自身氨基酸在微生物氨基酸脱羧酶催化下形成的生物胺, 该过程受(发酵时间、发酵温度、菌种添加量、添加剂等)影响[13-15]。适量生物胺的摄入不会对人体造成危害, 但是高浓度的生物胺摄入会对人体产生毒害作用, 过量的生物胺摄入可能会引发昏厥、休克, 严重的甚至会导致死亡[16-18], 生物胺类别及其毒性作用详见表1
生物胺中最常见且毒性最强的两种生物胺是组胺和酪胺。研究表明组胺与酪胺摄入过多会导致呕吐、引发血糖或血压失调、腹部痉挛等负面作用。我国规定高组胺鱼类组胺限量为400 mg/kg, 其他海水鱼类的组胺限量为200 mg/kg。
本研究采用高盐稀态发酵工艺, 对酱油在发酵过程中的生物胺含量进行分析, 重点探究了重要工艺参数(盐水浓度、发酵温度、酵母添加量)对酱油发酵过程中生物胺产生的影响, 以期降低酱油在发酵过程中生物胺的产生量, 提高酱油的食用安全性。
黄豆、小麦、米曲霉曲精等市售。
氯化钠、甲醛、氢氧化钠、乙腈、丙酮、乙醚、正丁醇、三氯甲烷、正已烷、正己烷(色谱纯)、酪胺盐酸盐(纯度≥98%)(上海阿拉丁生化科技股份有限公司); 乙酸、乙酸铵(色谱纯, 瑞春化学科技有限公司); 标准品: 组胺盐酸盐(纯度≥99%)、酪胺盐酸盐(纯度≥98%)、腐胺盐酸盐(纯度≥98%)、尸胺盐酸盐(纯度≥98%)、色胺盐酸盐(纯度≥99%)、亚精胺盐酸盐(纯度≥99%)、精胺盐酸盐(纯度≥99%)、β-苯乙胺(纯度≥98%)、章鱼胺盐酸盐(纯度≥98%)(上海麦克林生化科技有限公司)。
Agilent 1260高效液相色谱仪(美国安捷伦科技有限公司); ZHJH-C1109C超净工作台(苏州市金净设备科技有限公司); K1100全自动凯氏定氮仪、T960Basic全自动电位滴定仪(济南海能仪器股份有限公司); ZYDM-III-20T超纯水机(四川卓越水处理设备有限公司)。
本研究采用高盐稀态发酵, 工艺流程如图1所示。整个工艺主要分为两个阶段:
(1)制曲阶段。小麦进行炒制、粉碎、过筛, 大豆进行浸泡、蒸煮, 冷却至适宜的温度。小麦、大豆、米曲霉混合均匀进入圆盘进行制曲。制曲前半程曲料温度控制在28~33 ℃, 后面6~8 h温度控制在25~28 ℃, 总制曲时长为38~43 h, 制曲结束后进入发酵。
(2)发酵阶段。将曲料和盐水按一定比例混合, 盐水浓度15%、15 ℃冷温发酵一个月, 在自然升温至30 ℃后, 添加酵母菌继续进行发酵, 酵母添加量为0.05%。
冷温发酵结束后, 通过在保温夹层中通入冷水或者蒸汽进行升温或者降温的方式对酱油温度进行控制, 温度分别控制为20、30、40 ℃进行后续发酵。同时在发酵开始30、60、90、120、150、180 d时进行取样。取样时先通气搅拌10 min, 使得物料均一, 3个点随机取样500 mL, 使用80目滤布对样品进行过滤, 备用。检测滤后酱油的氨基酸态氮、全氮、总酸、生物胺等重要指标。
氨基酸态氮、总酸、可溶性无盐固形物、全氮参照GB/T 18186—2000《酿造酱油》测定。
总酸、氨基酸态氮的测定: 吸取20 mL 20倍稀释后的样品, 加60 mL超纯水, 采用0.05 mol/L氢氧化钠溶液滴至pH=8.2, 计算酱油中总酸的含量。继续加入10 mL甲醛溶液, 采用0.05 mol/L氢氧化钠溶液滴至pH=9.2, 计算酱油中氨基酸态氮的含量。
全氮的测定: 试样2 mL、4 g硫酸铜-硫酸钾混合试剂及10 mL硫酸混合后于凯氏消化管中消化, 消化完全后当消化管中溶液呈蓝绿色并澄清透明, 采用2%硼酸溶液吸收, 0.1 mol/L盐酸滴定, 计算酱油中的全氮含量。
生物胺的测定采用GB 5009.208—2016《食品安全国家标准 食品中生物胺的测定》。
准确量取1.0 mL样品, 依次加入250 μL 100 mg/L内标溶液(1,7-二氨基庚烷)、1 mL饱和碳酸氢钠溶液、100 μL氢氧化钠溶液(1 mol/L)、1 mL衍生试剂(丹磺酰氯)后置于60 ℃恒温水浴锅中衍生15 min, 取出后加入100 μL谷氨酸钠溶液, 混匀后60 ℃恒温反应15 min, 反应结束后冷却至室温, 再加入1 mL超纯水, 后40 ℃水浴下氮吹除去丙酮, 加入0.5 g氯化钠振荡至完全溶解, 再加入5 mL乙醚, 振荡摇匀后静置分层, 再萃取一次, 合并两次乙醚萃取液, 40 ℃水浴下氮气吹干。加入1 mL乙腈振荡混匀, 0.22 μm滤膜针头滤器过滤后进行测定, 色谱柱为C18柱, 紫外检测波长254 nm, 进样量20 μL, 柱温35 ℃, 流动相A为90%乙腈/10%(含0.1%乙酸的0.01 mol/L乙酸铵溶液), 流动相B为10%乙腈/90%(含0.1%乙酸的0.01 mol/L乙酸铵溶液), 流速0.8 mL/min。
研究表明酱油发酵过程中不同发酵时间、盐水浓度、温度、酵母添加量等对生物胺产生均有影响, 因此本研究考察不同时间(30、60、90、120、150、180 d)、盐水浓度(15%、20%、25%)、温度(20、30、40 ℃)、酵母添加量(0.05%、0.10%、0.15%)对酱油发酵过程中生物胺形成的影响。
每个样品设3个平行, 并使用SPSS 20.0进行统计学分析, P<0.05具有显著性差异; 再利用Origin 2024对实验数据和统计结果制图。
原料中蛋白质的分解程度可由酱油中全氮含量反映, 全氮指的是酱油中未分解的蛋白质、多肽、游离氨基酸等可溶性含氮化合物的总和[23-25]。其中氨基酸态氮是酱油重要鲜味指标, 同时氨基酸也是生物胺形成的前体物质, 其含量高低影响生物胺的生成[26-28]。酸性条件下微生物的生长受到抑制, 会降低酶类生成及酶活, 进而影响生物胺的产生。因此检测全氮、氨基酸态氮、总酸等理化指标。总酸作为酱油中的风味物质[29-32], 发酵前期总酸快速增长, 这是由于微生物将糖类与游离氨基酸转化为酸类。如图2(a)所示, 在前90 d, 总酸含量迅速增长至9.89 g/L。此后总酸增长速率减缓, 在180 d时, 总酸含量为11.47 g/L, 推测在发酵前期糖被微生物利用, 导致微生物代谢旺盛, 产生酸类物质, 导致酸含量迅速上升。发酵后期由于糖类减少微生物代谢减弱酸度变化缓慢。如图2(b)所示, 前期由于蛋白酶活性高, 原料中的蛋白质被迅速分解, 因此氨基酸态氮含量迅速上升。发酵后期, 总酸含量较高, 影响蛋白酶的活性, 同时酱油中还未分解的蛋白质含量较低, 因此氨基酸态氮的增长速率缓慢并逐渐趋于稳定, 在发酵时间为180 d时, 氨基酸态氮的含量为1.1 g/100 mL。如图2(c)所示, 随着发酵时间增长, 全氮的含量逐渐增加并在发酵后期维持基本不变, 当发酵时间为180 d时, 全氮的含量为1.55 g/100 mL。
在酱油的发酵过程中, 生物胺主要由氨基酸脱羧酶脱羧生成, 部分脂肪族生物胺由醛、酮的胺化和转氨化生成。
图3所示, 在由本研究发酵而成的酱油中的生物胺主要是尸胺、组胺、腐胺、酪胺这4种, 出峰时间分别是20.080、21.128、18.391、27.938 min, 其余色胺、苯乙胺、亚精胺、章鱼胺、精胺未在本研究发酵而成的酱油中检出。该结果与曾新安等[33]报道的结果类似, 其研究了16款市售酱油, 其中每款都含有酪胺和组胺, 37.5%含有尸胺, 93.75%含有腐胺。
生物胺的前体物质为氨基酸, 酱油发酵过程中的生物胺主要由微生物分泌的氨基酸脱羧酶催化氨基酸脱羧形成[14]。因此本研究考察了在发酵过程中生物胺随着发酵时间延长的变化规律。
图4(a)所示, 随着发酵时间的延长, 生物胺的含量逐渐上升, 发酵120 d后生物胺含量达到顶峰, 最高值为524.43 mg/L, 但随着发酵时间的延长, 生物胺的含量出现下降趋势, 因为在发酵过程中, 蛋白质逐渐被米曲霉分解成氨基酸, 随着氨基酸含量的增加, 微生物分泌的氨基酸脱羧酶作用于氨基酸不断产生生物胺, 导致生物胺的含量增加。但延长发酵时间, 生物胺含量出现下降的趋势, 这是由于在发酵的后期还原糖、前体氨基酸等被不断消耗, 易于生成生物胺的物质减少, 从而使得生物胺生成减少甚至停滞。同时由于长时间自然接种, 环境中某一些能够分解生物胺的微生物自发生长繁殖, 缓慢降低生物胺含量。进一步延长发酵时间生物胺含量可能会进一步降低, 但是经济成本较大, 因此发酵时间的研究选取180 d较为合适。如图4(b)所示, 在酱油的发酵过程中, 酪胺占总生物胺的50%左右, 组胺占总生物胺的25%, 尸胺占总生物胺的20%, 而腐胺的占比最小, 约5%。该结论与报道一致, 酪胺是酱油中最普遍且含量最高的生物胺[34]
高盐水浓度会造成高渗透压环境, 对微生物的生长具有抑制效果。为了探究盐水浓度对生物胺生成的影响。检测了不同盐水浓度(15%、20%、25%)下, 发酵过程中生物胺含量的变化规律。
图5(a)所示, 随着发酵时间的延长, 不同盐水浓度的酱油在发酵过程中的生物胺的变化情况均为先增加后下降, 不同盐水浓度的酱油在发酵过程中生物胺含量的峰值不一样, 15%、20%盐水浓度的酱油生物胺含量在120 d达到峰值, 分别为524.43 mg/L、478.02 mg/L; 25%盐水浓度的酱油生物胺含量在150 d达到峰值, 为449.31 mg/L。其中在酱油发酵180 d后, 添加15%盐水浓度的酱油生物胺含量最高, 添加25%盐水浓度的酱油中生物胺含量最低, 为376.41 mg/L, 因为高浓度的盐溶液可以抑制大量杂菌的繁殖, 降低氨基酸脱羧酶的活性, 该实验结果与报道一致[34]。因此推断高盐环境导致微生物生长受到抑制, 导致25%盐水浓度的酱油中生物胺的含量最少且峰值出现时间后延。但不同盐水浓度对酱油中生物胺组成的占比影响不大, 都存在同一种现象, 酪胺占总生物胺的比例最大, 其次是组胺, 如图5(b)所示。
发酵温度对生物胺的生成具有较大影响。为了探究发酵温度对生物胺产生的影响, 考察了不同发酵温度(20、30、40 ℃)下, 酱油发酵过程中生物胺的变化规律。
图6(a)所示, 生物胺在不同发酵温度酱油中的含量变化均为先增加后减少。20、30、40 ℃发酵温度下酱油中生物胺峰值依次为507.43、554.43、475.21 mg/L, 其中发酵温度为30 ℃的酱油中生物胺含量最高。在40 ℃的发酵温度下, 在90 d生物胺的含量就达到了峰值, 较高的发酵温度促使蛋白质的水解和脱羧酶反应速率增加, 生物胺生成趋势变快。但是随着发酵时间的延长, 较高的温度导致酱油中水分的蒸发速度相较低温时更为迅速。因此酱油中盐水浓度不断上升, 渗透压不断增加, 从而抑制氨基酸脱羧酶的产生, 进一步抑制了生物胺的产生, 在发酵结束后, 40 ℃发酵温度的酱油中生物胺含量为368.9 mg/L。此外较高的温度促使美拉德反应的产生, 导致氨基酸与还原糖结合, 氨基酸含量下降, 此实验结果与报道一致[35]。同时, 如图6(b)所示, 在发酵120 d后, 20 ℃下发酵酱油的生物胺中酪胺的含量明显高于30 ℃、40 ℃下发酵的酱油, 而30 ℃下发酵酱油的生物胺中组胺的含量高于20 ℃、40 ℃下发酵的酱油。
酱油发酵阶段加入的酵母在一定程度上改变了酱油中各种物质的溶解度和浓度。同时, 也会对微生物的生长、代谢产生影响。为了探究酵母添加量对生物胺产生的影响, 考察了不同酵母添加量(0.05%、0.10%、0.15%)下, 酱油发酵过程中生物胺的变化规律。
图7(a)所示, 随着发酵时间的延长, 不同酵母添加量的酱油在发酵过程中总生物胺的产生情况均为先增加后下降。0.05%、0.10%、0.15%酵母添加量发酵的酱油中生物胺含量峰值依次为524.43、551.26、595.43 mg/L, 其中0.15%酵母添加量的酱油中生物胺含量最高, 0.05%酵母添加量的酱油中生物胺含量最低, 为403.45 mg/L。生物胺含量随着酵母接种量的增加而增加, 可能是酵母菌的添加导致酱油中氨基酸脱羧酶增加, 该结论与朱子桂[35]研究的酵母添加量对酱油中生物胺的影响结论一致。同时, 如图7(b)所示, 酵母添加量对酱油中生物胺组成和占比影响不大。
由上述结果可得, 随发酵时间延长, 生物胺的含量呈现先增加后降低的趋势。可能是发酵体系中产生物胺的微生物生长受到抑制导致氨基酸脱羧酶催化活性下降。而且在长期发酵过程中自然接种一部分能够降解生物胺的微生物, 并在发酵后期成为优势菌群从而使得生物胺含量下降[36]
通过发酵温度实验发现, 当温度较高时, 生物胺生成速率较快, 但生物胺总量变低, 提示可以通过提升温度来促进发酵进程, 抑制生物胺的生成。值得注意的是, 较高的发酵温度虽然可以降低生物胺的含量, 但由于其具有选择性, 可能会导致菌种相对单一, 酱油风味受到影响, 同时酱油所能存储的时间也相对较短。
此外通过盐水浓度实验表明, 适量提高盐水浓度, 能够显著性抑制生物胺的产生, 因为高盐水浓度带来的高渗透压使得生物胺的生成受到更为明显的抑制, 推测为高浓度渗透压破坏了酶的蛋白结构, 抑制了微生物的生长, 导致氨基酸脱羧酶的催化作用受到了抑制, 因此在高浓度盐溶液下, 酱油中的生物胺含量较少。
酵母添加量对生物胺的产生有重要影响, 实验结果表明, 在超过一定限度的酵母接种情况下, 酵母菌生长较为迅速, 导致生物胺在前期过程中大量积聚。同时酵母过度繁殖导致酱油中糖分被应用于微生物生长, 其产生醇相对较少, 因此风味较为寡淡。较低的酵母接种量下生物胺总量最低, 但酵母数量较少导致其产生乙醇能力受到抑制, 也会造成风味比较寡淡的现象。
本研究探索了在不同温度、发酵时长、盐水浓度、酵母接种量对于生物胺产生的影响。发现在40 ℃的温度下, 酱油发酵结束后的生物胺含量最低, 为368.9 mg/L; 在盐水浓度为25%时, 酱油发酵结束后的生物胺含量最低, 为376.41 mg/L; 酵母添加量为0.05%时, 酱油发酵结束后的生物胺含量最低, 为403.45 mg/L。生物胺作为酱油发酵过程中的有害物质, 其含量过高会对人体健康产生一定影响, 通过改变盐水浓度、发酵温度和酵母添加量, 可以有效抑制酱油发酵过程中生物胺的形成, 从而有效提高酱油的安全性。但本研究中所述实验仍有一定不足, 即所设定梯度范围相对较大, 且只对酱油发酵过程中部分因素进行了探究, 后续可复合添加菌种、改变酱油发酵的pH等多个方面进行优化, 提升酱油的品质。还可以对酱油发酵过程中降解生物胺的微生物进行筛选研究, 减少酱油发酵过程中的生物胺的生成。
  • 泸州市科技计划项目(2023NYF089)
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2025年第16卷第7期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241030009
  • 接收时间:2024-10-30
  • 首发时间:2025-07-19
  • 出版时间:2025-04-15
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  • 收稿日期:2024-10-30
基金
泸州市科技计划项目(2023NYF089)
作者信息
    1.四川元景达食品有限公司, 泸州 646000
    2.泸州老窖集团有限责任公司, 泸州 646000

通讯作者:

* 傅其旭(1985—), 男, 工程师, 主要研究方向为酒类及调味品酿造。E-mail:
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https://castjournals.cast.org.cn/joweb/spaq/CN/10.19812/j.cnki.jfsq11-5956/ts.20241030009
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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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