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Article(id=1151437196268548396, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151437189243089177, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241222001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1734796800000, receivedDateStr=2024-12-22, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752453620259, onlineDateStr=2025-07-14, pubDate=1749916800000, pubDateStr=2025-06-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752453620259, onlineIssueDateStr=2025-07-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752453620259, creator=13701087609, updateTime=1752453620259, 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=179, endPage=185, ext={EN=ArticleExt(id=1151895327800963092, articleId=1151437196268548396, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Study on acute toxicity and genetic toxicity of γ-polyglutamic acid, 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 evaluate the acute toxicity and genetic toxicity of γ-polyglutamic acid (γ-PGA). Methods Acute toxicity was evaluated by oral gavage of γ-PGA at 10000 mg/(kg·bw) to rats and mice using the limit test method, with observations of toxic signs and mortality; the 30 hour double gavage method followed by microscopic examination to quantify the frequency of micronucleated polychromatic erythrocytes in mouse bone marrow was employed to detect the micronucleus induction of γ-PGA; the plate incorporation method was utilized to enumerate revertant colonies of Salmonella typhimurium strains (TA97a, TA98, TA100, TA102 and TA1535) to assess the mutagenic potential of γ-PGA; a continuous 5 day gavage followed by microscopic analysis was conducted to analyze the number and frequency of chromosomal structural aberrations in mouse spermatocytes to evaluate the chromosomal aberration induction of γ-PGA. Results The maximum tolerated dose (MTD) of γ-PGA via acute oral administration to both male and female mice and rats was greater than 10000 mg/(kg·bw). At a dose of 5000 mg/(kg·bw), γ-PGA did not significantly increase micronucleated polychromatic erythrocytes in the bone marrow of either male or female mice (P>0.05), nor did it induce chromosomal aberrations in primary spermatocytes of mice (P>0.05). Additionally, at a dose of 5000 μg per plate, no significant mutagenic activity was detected in the standard tested strains (TA97a, TA98, TA100, TA102 and TA1535), regardless of S9 activation. Conclusion Under the experimental conditions of this study, γ-PGA is classified as practically non-toxic and non-genotoxic, providing a toxicological basis for its development and application in the food industry.

, correspAuthors=Ping YU, 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=Yue ZHAO, Rong ZHAO, Zhong-Ming LV, Ping YU), CN=ArticleExt(id=1151895328186839062, articleId=1151437196268548396, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=γ-聚谷氨酸急性毒性和遗传毒性研究, 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 评价γ-聚谷氨酸(γ-polyglutamic acid, γ-PGA)急性毒性和遗传毒性。方法 采用限量法, 分别以10000 mg/(kg·bw)的剂量γ-PGA灌胃给予大鼠和小鼠, 通过观察动物中毒表现及死亡情况, 检测其急性毒性; 采用30 h两次灌胃法, 镜检计数小鼠骨髓有微核嗜多染红细胞频率, 检测γ-PGA致微核作用; 采用平板掺入法, 计数TA97a、TA98、TA100、TA102和TA1535菌株的回变菌落数, 检测γ-PGA诱导基因突变活性; 采用连续5 d灌胃法, 镜检计数小鼠精母细胞染色体结构畸变数目及频率, 检测γ-PGA致染色体畸变作用。结果 γ-PGA对雌雄小鼠和大鼠的急性经口最大耐受剂量(maximum tolerated dose, MTD)均大于10000 mg/(kg· bw); 剂量达5000 mg/(kg·bw), 未见对雌雄小鼠骨髓嗜多染红细胞的致微核作用(P>0.05), 以及未见对小鼠初级精母细胞的致染色体畸变作用(P>0.05); 剂量达5000 μg/皿, 有无S9条件下, 均未对标准测试菌株TA97a、TA98、TA100、TA102和TA1535表现出明显的诱变活性。结论 本研究条件下, γ-PGA属于实际无毒级且无遗传毒性, 为其在食品领域的开发和应用提供了毒理学依据。

, correspAuthors=俞萍, authorNote=null, correspAuthorsNote=
* 俞萍(1973—), 女, 硕士, 主任医师, 主要研究方向为食品和新食品原料毒理学安全性评价与风险评估。E-mail:
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赵悦(1991—), 女, 硕士, 主管技师, 主要研究方向为食品和保健食品毒理学检测与功能评价。E-mail:

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赵悦(1991—), 女, 硕士, 主管技师, 主要研究方向为食品和保健食品毒理学检测与功能评价。E-mail:

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赵悦(1991—), 女, 硕士, 主管技师, 主要研究方向为食品和保健食品毒理学检测与功能评价。E-mail:

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Journal of Food Safety & Quality, 2021, 12(10): 3919-3926., articleTitle=Safety evaluation of edible dock as a new food raw material, refAbstract=null)], funds=null, companyList=[AuthorCompany(id=1167030577162105499, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, xref=null, ext=[AuthorCompanyExt(id=1167030577170494108, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, companyId=1167030577162105499, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Institute of Toxicology and Risk Assessment, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China), AuthorCompanyExt(id=1167030577174688413, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, companyId=1167030577162105499, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=江苏省疾病预防控制中心毒理与风险评估研究所, 南京 210009)])], figs=[ArticleFig(id=1167030579146011320, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 1, caption=

Effects of γ-PGA on the body weight of mice

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 动物数/只 初始体重/g 一周体重/g 二周体重/g
雌 10 20.2±0.7 25.1±1.1 28.6±1.3
雄 10 20.9±1.0 28.1±1.3 33.9±1.5
), ArticleFig(id=1167030579213120185, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表1, caption=

γ-PGA对小鼠体重的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 动物数/只 初始体重/g 一周体重/g 二周体重/g
雌 10 20.2±0.7 25.1±1.1 28.6±1.3
雄 10 20.9±1.0 28.1±1.3 33.9±1.5
), ArticleFig(id=1167030579259257530, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 2, caption=

Results of acute oral toxicity test in mice

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 剂量/[mg/(kg·bw)] 动物数
/只		 出现中毒表现动物数/只 MTD
/[mg/(kg·bw)]
雌 10000 10 0 >10000
雄 10000 10 0 >10000
), ArticleFig(id=1167030579317977787, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表2, caption=

小鼠急性经口毒性试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 剂量/[mg/(kg·bw)] 动物数
/只		 出现中毒表现动物数/只 MTD
/[mg/(kg·bw)]
雌 10000 10 0 >10000
雄 10000 10 0 >10000
), ArticleFig(id=1167030579410252476, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 3, caption=

Effects of γ-PGA on the body weight of rat

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 动物数/只 初始体重/g 一周体重/g 二周体重/g
雌 10 191±6 225±8 257±12
雄 10 202±13 258±20 316±29
), ArticleFig(id=1167030579468972733, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表3, caption=

γ-PGA对大鼠体重的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 动物数/只 初始体重/g 一周体重/g 二周体重/g
雌 10 191±6 225±8 257±12
雄 10 202±13 258±20 316±29
), ArticleFig(id=1167030579548664510, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 4, caption=

Results of acute oral toxicity test in rat

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 剂量/[mg/(kg·bw)] 动物数
/只		 出现中毒表现动物数/只 MTD
/[mg/(kg·bw)]
雌 10000 10 10 >10000
雄 10000 10 10 >10000
), ArticleFig(id=1167030579619967679, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表4, caption=

大鼠急性经口毒性试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 剂量/[mg/(kg·bw)] 动物数
/只		 出现中毒表现动物数/只 MTD
/[mg/(kg·bw)]
雌 10000 10 10 >10000
雄 10000 10 10 >10000
), ArticleFig(id=1167030579691270848, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 5, caption=

Micronucleus test results of bone marrow erythrocytes in mice

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 组别 剂量
/[mg/(kg·bw)]		 动物数
/只		 PCE数
/个		 含微核
PCE数/个		 PCE/(PCE+NCE)
/%		 含微核PCE率
/‰
雌性 溶剂对照 0 5 10000 18 50.0±1.5 1.8±0.6
低剂量组 1250 5 10000 15 50.3±1.3 1.5±0.7
中剂量组 2500 5 10000 19 50.1±1.7 1.9±0.7
高剂量组 5000 5 10000 15 50.8±1.1 1.5±0.8
阳性对照 40 5 10000 266 49.4±1.3 26.6±6.2*
雄性 溶剂对照 0 5 10000 17 50.1±1.1 1.7±0.4
低剂量组 1250 5 10000 14 49.8±1.8 1.4±0.4
中剂量组 2500 5 10000 17 50.1±1.6 1.7±1.0
高剂量组 5000 5 10000 16 50.2±1.1 1.6±0.4
阳性对照 40 5 10000 257 49.7±1.0 25.7±3.7*
), ArticleFig(id=1167030579770962625, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表5, caption=

小鼠骨髓红细胞微核试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
性别 组别 剂量
/[mg/(kg·bw)]		 动物数
/只		 PCE数
/个		 含微核
PCE数/个		 PCE/(PCE+NCE)
/%		 含微核PCE率
/‰
雌性 溶剂对照 0 5 10000 18 50.0±1.5 1.8±0.6
低剂量组 1250 5 10000 15 50.3±1.3 1.5±0.7
中剂量组 2500 5 10000 19 50.1±1.7 1.9±0.7
高剂量组 5000 5 10000 15 50.8±1.1 1.5±0.8
阳性对照 40 5 10000 266 49.4±1.3 26.6±6.2*
雄性 溶剂对照 0 5 10000 17 50.1±1.1 1.7±0.4
低剂量组 1250 5 10000 14 49.8±1.8 1.4±0.4
中剂量组 2500 5 10000 17 50.1±1.6 1.7±1.0
高剂量组 5000 5 10000 16 50.2±1.1 1.6±0.4
阳性对照 40 5 10000 257 49.7±1.0 25.7±3.7*
), ArticleFig(id=1167030579838071490, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 6, caption=

Results of the first bacterial reverse mutation test (n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/(μg/皿)		 TA97a TA98 TA100 TA102 TA1535
+S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9
自发回变 / 105±15 101±6 36±1 32±2 83±10 102±6 311±7 265±17 21±4 15±3
二甲基亚砜 / 105±11 99±13 33±4 31±2 87±7 102±15 313±6 263±8 20±3 17±3
纯水 / 106±9 97±7 34±5 31±4 88±9 99±8 314±6 271±13 21±4 16±4
γ-PGA 8 101±4 92±13 33±2 29±1 89±5 101±4 294±20 267±7 19±3 18±2
40 98±11 93±14 37±5 33±6 84±8 97±14 317±4 269±3 19±4 16±3
200 103±11 92±8 33±2 31±1 92±13 108±8 313±8 271±5 20±5 17±4
1000 104±10 94±14 36±1 34±2 89±8 106±4 304±12 276±13 19±3 18±3
5000 106±16 93±14 32±4 29±1 90±11 99±7 313±8 268±15 21±5 16±4
敌克松 50 / 1232±87* / 1270±131* / / / 1261±113* / /
叠氮钠 1.5 / / / / / 1038±90* / / / 414±97*
2-氨基芴 10 1340±109* / 1075±91* / 1372±74* / / / / /
1,8-二羟基蒽醌 50 / / / / / / 857±91* / / /
环磷酰胺 20 / / / / / / / / 215±35* /
), ArticleFig(id=1167030579896791747, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表6, caption=

第一次细菌回复突变试验结果(n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/(μg/皿)		 TA97a TA98 TA100 TA102 TA1535
+S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9
自发回变 / 105±15 101±6 36±1 32±2 83±10 102±6 311±7 265±17 21±4 15±3
二甲基亚砜 / 105±11 99±13 33±4 31±2 87±7 102±15 313±6 263±8 20±3 17±3
纯水 / 106±9 97±7 34±5 31±4 88±9 99±8 314±6 271±13 21±4 16±4
γ-PGA 8 101±4 92±13 33±2 29±1 89±5 101±4 294±20 267±7 19±3 18±2
40 98±11 93±14 37±5 33±6 84±8 97±14 317±4 269±3 19±4 16±3
200 103±11 92±8 33±2 31±1 92±13 108±8 313±8 271±5 20±5 17±4
1000 104±10 94±14 36±1 34±2 89±8 106±4 304±12 276±13 19±3 18±3
5000 106±16 93±14 32±4 29±1 90±11 99±7 313±8 268±15 21±5 16±4
敌克松 50 / 1232±87* / 1270±131* / / / 1261±113* / /
叠氮钠 1.5 / / / / / 1038±90* / / / 414±97*
2-氨基芴 10 1340±109* / 1075±91* / 1372±74* / / / / /
1,8-二羟基蒽醌 50 / / / / / / 857±91* / / /
环磷酰胺 20 / / / / / / / / 215±35* /
), ArticleFig(id=1167030579972289220, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 7, caption=

Results of the second bacterial reverse mutation test (n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/(μg/皿)		 TA97a TA98 TA100 TA102 TA1535
+S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9
自发回变 / 103±13 101±5 34±3 32±4 83±8 104±2 309±12 270±11 20±4 19±5
二甲基
亚砜		 / 99±7 96±6 33±5 31±3 89±5 97±6 306±22 269±17 19±2 17±4
纯水 / 97±7 92±7 34±1 31±2 79±10 90±12 312±14 288±15 17±3 16±2
γ-PGA 19.5 103±2 95±12 33±3 32±4 81±12 99±11 312±10 270±9 21±4 16±3
78.1 104±13 96±9 35±4 33±3 88±3 98±12 312±7 265±10 20±5 18±3
312.5 94±11 87±10 31±5 30±2 87±11 101±7 307±17 269±10 21±4 19±5
1250 101±11 90±5 35±3 31±2 92±3 103±8 310±8 265±12 19±5 17±1
5000 96±13 97±9 33±2 32±1 84±3 101±9 311±12 268±14 20±3 17±4
敌克松 50 / 1276±131* / 1263±107* / / / 1228±42* / /
叠氮钠 1.5 / / / / / 1071±129* / / / 443±93*
2-氨基芴 10 1284±45* / 1098±44* / 1397±90* / / / / /
1,8-二羟基蒽醌 50 / / / / / / 802±97* / / /
环磷酰胺 20 / / / / / / / / 177±38* /
), ArticleFig(id=1167030580064563909, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表7, caption=

第二次细菌回复突变试验结果(n=3)

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/(μg/皿)		 TA97a TA98 TA100 TA102 TA1535
+S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9 +S9 -S9
自发回变 / 103±13 101±5 34±3 32±4 83±8 104±2 309±12 270±11 20±4 19±5
二甲基
亚砜		 / 99±7 96±6 33±5 31±3 89±5 97±6 306±22 269±17 19±2 17±4
纯水 / 97±7 92±7 34±1 31±2 79±10 90±12 312±14 288±15 17±3 16±2
γ-PGA 19.5 103±2 95±12 33±3 32±4 81±12 99±11 312±10 270±9 21±4 16±3
78.1 104±13 96±9 35±4 33±3 88±3 98±12 312±7 265±10 20±5 18±3
312.5 94±11 87±10 31±5 30±2 87±11 101±7 307±17 269±10 21±4 19±5
1250 101±11 90±5 35±3 31±2 92±3 103±8 310±8 265±12 19±5 17±1
5000 96±13 97±9 33±2 32±1 84±3 101±9 311±12 268±14 20±3 17±4
敌克松 50 / 1276±131* / 1263±107* / / / 1228±42* / /
叠氮钠 1.5 / / / / / 1071±129* / / / 443±93*
2-氨基芴 10 1284±45* / 1098±44* / 1397±90* / / / / /
1,8-二羟基蒽醌 50 / / / / / / 802±97* / / /
环磷酰胺 20 / / / / / / / / 177±38* /
), ArticleFig(id=1167030580144255686, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=EN, label=Table 8, caption=

Result of chromosome aberration test in mice spermatocytes

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/[mg/(kg·bw)]		 动物数
/只		 观察
细胞数/个		 裂隙/个 性染色体
单价体/%		 常染色体
单价体/%		 染色体畸变类型 畸变
细胞数/个		 畸变
细胞率/%
断片/个 易位/个 微小体/个
阴性对照 0 5 500 0 3.2±1.3 1.6±1.1 3 0 0 3 0.6±0.9
低剂量组 1250 5 500 1 3.4±1.1 1.6±1.5 2 0 0 2 0.4±0.5
中剂量组 2500 5 500 0 3.2±1.9 1.6±0.9 2 0 2 4 0.8±0.8
高剂量组 5000 5 500 0 3.0±1.6 1.4±1.1 2 0 0 2 0.4±0.5
阳性对照 2 5 500 7 8.2±1.9* 6.0±1.6* 44 0 4 47 9.4±2.1*
), ArticleFig(id=1167030580244918983, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437196268548396, language=CN, label=表8, caption=

小鼠精母细胞染色体畸变试验结果

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 剂量
/[mg/(kg·bw)]		 动物数
/只		 观察
细胞数/个		 裂隙/个 性染色体
单价体/%		 常染色体
单价体/%		 染色体畸变类型 畸变
细胞数/个		 畸变
细胞率/%
断片/个 易位/个 微小体/个
阴性对照 0 5 500 0 3.2±1.3 1.6±1.1 3 0 0 3 0.6±0.9
低剂量组 1250 5 500 1 3.4±1.1 1.6±1.5 2 0 0 2 0.4±0.5
中剂量组 2500 5 500 0 3.2±1.9 1.6±0.9 2 0 2 4 0.8±0.8
高剂量组 5000 5 500 0 3.0±1.6 1.4±1.1 2 0 0 2 0.4±0.5
阳性对照 2 5 500 7 8.2±1.9* 6.0±1.6* 44 0 4 47 9.4±2.1*
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γ-聚谷氨酸急性毒性和遗传毒性研究
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赵悦 , 赵荣 , 吕中明 , 俞萍 *
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(11): 179-185
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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(11): 179-185
γ-聚谷氨酸急性毒性和遗传毒性研究
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赵悦 , 赵荣, 吕中明, 俞萍*
作者信息
  • 江苏省疾病预防控制中心毒理与风险评估研究所, 南京 210009

    • 赵悦(1991—), 女, 硕士, 主管技师, 主要研究方向为食品和保健食品毒理学检测与功能评价。E-mail:

通讯作者:

* 俞萍(1973—), 女, 硕士, 主任医师, 主要研究方向为食品和新食品原料毒理学安全性评价与风险评估。E-mail:
Study on acute toxicity and genetic toxicity of γ-polyglutamic acid
Yue ZHAO , Rong ZHAO, Zhong-Ming LV, Ping YU*
Affiliations
  • Institute of Toxicology and Risk Assessment, Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China
出版时间: 2025-06-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241222001
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摘要
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目的 评价γ-聚谷氨酸(γ-polyglutamic acid, γ-PGA)急性毒性和遗传毒性。方法 采用限量法, 分别以10000 mg/(kg·bw)的剂量γ-PGA灌胃给予大鼠和小鼠, 通过观察动物中毒表现及死亡情况, 检测其急性毒性; 采用30 h两次灌胃法, 镜检计数小鼠骨髓有微核嗜多染红细胞频率, 检测γ-PGA致微核作用; 采用平板掺入法, 计数TA97a、TA98、TA100、TA102和TA1535菌株的回变菌落数, 检测γ-PGA诱导基因突变活性; 采用连续5 d灌胃法, 镜检计数小鼠精母细胞染色体结构畸变数目及频率, 检测γ-PGA致染色体畸变作用。结果 γ-PGA对雌雄小鼠和大鼠的急性经口最大耐受剂量(maximum tolerated dose, MTD)均大于10000 mg/(kg· bw); 剂量达5000 mg/(kg·bw), 未见对雌雄小鼠骨髓嗜多染红细胞的致微核作用(P>0.05), 以及未见对小鼠初级精母细胞的致染色体畸变作用(P>0.05); 剂量达5000 μg/皿, 有无S9条件下, 均未对标准测试菌株TA97a、TA98、TA100、TA102和TA1535表现出明显的诱变活性。结论 本研究条件下, γ-PGA属于实际无毒级且无遗传毒性, 为其在食品领域的开发和应用提供了毒理学依据。

γ-聚谷氨酸  /  急性毒性  /  遗传毒性

Objective To evaluate the acute toxicity and genetic toxicity of γ-polyglutamic acid (γ-PGA). Methods Acute toxicity was evaluated by oral gavage of γ-PGA at 10000 mg/(kg·bw) to rats and mice using the limit test method, with observations of toxic signs and mortality; the 30 hour double gavage method followed by microscopic examination to quantify the frequency of micronucleated polychromatic erythrocytes in mouse bone marrow was employed to detect the micronucleus induction of γ-PGA; the plate incorporation method was utilized to enumerate revertant colonies of Salmonella typhimurium strains (TA97a, TA98, TA100, TA102 and TA1535) to assess the mutagenic potential of γ-PGA; a continuous 5 day gavage followed by microscopic analysis was conducted to analyze the number and frequency of chromosomal structural aberrations in mouse spermatocytes to evaluate the chromosomal aberration induction of γ-PGA. Results The maximum tolerated dose (MTD) of γ-PGA via acute oral administration to both male and female mice and rats was greater than 10000 mg/(kg·bw). At a dose of 5000 mg/(kg·bw), γ-PGA did not significantly increase micronucleated polychromatic erythrocytes in the bone marrow of either male or female mice (P>0.05), nor did it induce chromosomal aberrations in primary spermatocytes of mice (P>0.05). Additionally, at a dose of 5000 μg per plate, no significant mutagenic activity was detected in the standard tested strains (TA97a, TA98, TA100, TA102 and TA1535), regardless of S9 activation. Conclusion Under the experimental conditions of this study, γ-PGA is classified as practically non-toxic and non-genotoxic, providing a toxicological basis for its development and application in the food industry.

γ-polyglutamic acid  /  acute toxicity  /  genetic toxicity
赵悦, 赵荣, 吕中明, 俞萍. γ-聚谷氨酸急性毒性和遗传毒性研究. 食品安全质量检测学报, 2025 , 16 (11) : 179 -185 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241222001
Yue ZHAO, Rong ZHAO, Zhong-Ming LV, Ping YU. Study on acute toxicity and genetic toxicity of γ-polyglutamic acid[J]. Journal of Food Safety & Quality, 2025 , 16 (11) : 179 -185 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241222001
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0 引言
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γ-聚谷氨酸(γ-polyglutamic acid, γ-PGA), 又称多聚谷氨酸, 是由D-谷氨酸和/或L-谷氨酸单体聚合而成的直链型生物高分子物质[1], 最初在炭疽芽孢杆菌的荚膜中被发现, 同时也存在于日本传统发酵食品纳豆黏液中[2]。目前, 以枯草芽孢杆菌为主要生产菌的微生物发酵法是其主要生产方式[3]。γ-PGA具有优良的水溶性、保水性、生物降解性、生物相容性等特性, 在食品、化妆品、农业、医药等领域均展现出了极大的应用潜力[4-5]。特别是在食品工业中, γ-PGA不仅可以作为绿色环保的食品包装材料[6], 还可以作为冻干保护剂有效延长食品保质期[7-8], 作为掩味剂、增稠剂和稳定剂等改善食品品质[9]。此外, γ-PGA还具有多种健康益处, 如: 促进肠道中钙和维生素的吸收[10-11]、改善肠道菌群健康[12]等。动物试验进一步表明, γ-PGA还具有抗氧化[13]、降血脂[14]、降血糖[15-16]、降胆固醇[17]等多种功效。
张彩霞等[18]依据GB 15193.13—2003《30天和90天喂养试验》, 采用γ-PGA掺入饲料的方式, 以5000~ 15000 mg/(kg·bw)剂量对大鼠进行为期30 d的亚急性毒性试验。研究结果显示, γ-PGA会显著抑制雌、雄大鼠日增重, 并导致雌、雄大鼠肝脏和肾脏重量及雄性大鼠睾丸重量与对照组相比出现显著性差异。这些结果提示, γ-PGA可能对动物的生长发育及特定器官功能产生潜在影响。因此, 本研究依据现行2014版《食品安全国家标准》, 系统评价了γ-PGA的急性经口毒性, 并通过小鼠骨髓细胞微核试验、细菌回复突变试验和小鼠精母细胞染色体畸变试验, 对其潜在遗传毒性进行了全面评价, 旨在为γ-PGA的长期毒性研究及其在食品领域的安全应用和开发提供毒理学依据。
1 材料与方法
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1.1 材料与试剂
γ-PGA: 南京某生物科技股份有限公司, 白色粉状, 阴凉、干燥处保存, 保存期限为2年。
环磷酰胺(纯度≥97.0%)、1,8-二羟基蒽醌(纯度≥95.5%)(美国Sigma公司); 敌克松(纯度≥95.0%, 美国AccuStandard公司); 叠氮钠(纯度≥99.5%, 浙江东阳市天宇化工有限公司); 2-氨基芴(纯度≥97.5%, 瑞士Fluka AG公司); 丝裂霉素C(纯度>98%, 日本东京化成工业株式会社); 秋水仙素(纯度≥98%)、柠檬酸三钠、甲醇、冰乙酸(分析纯)(国药集团化学试剂有限公司); 小牛血清(美国Gibco公司); Giemsa染液(上海碧云天生物技术股份有限公司); 纯水(Ⅰ级)。
TA97a、TA98、TA100、TA102和TA1535[19]组氨酸缺陷型的鼠伤寒沙门氏菌株(活菌浓度109个/mL以上, 美国Moltox公司), 经生物学鉴定符合要求, 各菌株于37 ℃条件下增菌培养过夜; S9大鼠肝匀浆(纯度10%, 江苏齐氏生物科技有限公司), 由β-萘黄酮、苯巴比妥联合诱导, 合格证号: 23FS016D, 代谢活化系统中S9浓度为10%。
1.2 仪器与设备
PANNORAMIC MIDI数字切片扫描系统(匈牙利3DHIESTECH公司); Axiostar plus生物显微镜[蔡司光学仪器(上海)国际贸易有限公司]; PL203型电子天平[精度0.1 mg, 梅特勒-托利多仪器(上海)有限公司]; JJ500型电子秤(精度10 mg, 常熟市双杰测试仪器厂); DS-671电子秤(精度1 g, 上海寺冈电子有限公司); Airstream Ⅱ级 A2型生物安全柜(新加坡艺思高科技有限公司); LRH-400A生化培养箱(北京泰宏君仪器有限公司); TOMY SX-700快速自动高压灭菌器[天美仪拓实验室设备(上海)有限公司]。
1.3 试验对象
SPF级ICR小鼠(合格证号: 20220004024774), SPF级SD大鼠(合格证号: 20220004024773), 均由上海斯莱克试验动物有限责任公司提供, 动物生产许可证号: SCXK(沪)2022-0004。动物饲养在屏障环境中, 环境设施使用许可证号: SYXK(苏)2022-0034, 环境温度: 20~24 ℃, 相对湿度: 40%~70%, 12/12 h昼夜明暗交替。试验期间大、小鼠自由饮用灭菌水、摄食灭菌维持饲料, 维持饲料来源及生产许可证号: 江苏省协同医药生物工程有限责任公司, 苏饲证(2019)01008。
1.4 试验方法
1.4.1 试验设计依据
γ-PGA属于食品添加剂类别, 依据GB 15193.1—2014《食品安全国家标准 食品安全性毒理学评价程序》中“食品添加剂毒理学评价”的要求, 开展急性经口毒性试验、遗传毒性试验(包括细菌回复突变试验、小鼠骨髓细胞微核试验和小鼠精母细胞染色体畸变试验)毒理学安全性评价。
1.4.2 急性经口毒性试验(小鼠)
选用SPF级健康ICR小鼠20只, 雌雄各半, 初始体重18~22 g。采用限量法, 设计剂量为10000 mg/(kg·bw), 即称取γ-PGA 10000 mg, 加纯水定容至40 mL作为受试物。小鼠禁食6 h后, 将受试物分两次灌胃给予, 每次灌胃容量均为20 mL/(kg·bw), 时间间隔4 h, 灌胃后2 h给食, 试验期间均不限制饮水, 观察期14 d, 记录小鼠中毒表现及死亡情况。
1.4.3 急性经口毒性试验(大鼠)
选用SPF级健康SD大鼠20只, 雌雄各半, 初始体重180~220 g。采用限量法, 设计剂量为10000 mg/(kg·bw), 即称取γ-PGA 30000 mg, 加纯水定容至120 mL作为受试物。大鼠禁食过夜后, 将受试物分两次灌胃给予, 每次灌胃容量均为20 mL/(kg·bw), 时间间隔4 h, 灌胃后4 h给食, 试验期间均不限制饮水, 观察期14 d, 记录大鼠中毒表现及死亡情况。
1.4.4 小鼠骨髓细胞微核试验
选用SPF级健康ICR小鼠50只, 雌雄各半, 初始体重25~30 g。将小鼠分性别按体重随机分入3个样品剂量组[5000、2500、1250 mg/(kg·bw)]、1个溶剂对照组(纯水)和1个阳性对照组[环磷酰胺, 40 mg/(kg·bw)], 每组10只, 雌雄各半。采用30 h两次灌胃法, 灌胃间隔时间24 h, 每次灌胃容量均为20 mL/(kg·bw), 第二次灌胃后6 h取小鼠股骨骨髓悬于小牛血清中直接涂片, 经甲醇固定、Giemsa液染色后, 每只动物镜检嗜多染红细胞(polychromatic erythrocyte, PCE) 2000个, 计数含微核的PCE数。每只动物观察200个PCE的同时计数所见到的正染红细胞(normochromatic erythrocyte, NCE)数, 计算PCE数在总红细胞(PCE+NCE)数中的比例。
1.4.5 细菌回复突变试验
设5个样品剂量组, 称取γ-PGA 500 mg用纯水定容至10 mL, 经121 ℃、20 min高压灭菌作为最高剂量组受试物(5000 μg/皿)。无菌环境条件下准确量取最高剂量组受试物, 用灭菌纯水依次连续作5倍稀释配成其他剂量组受试液(1000、200、40、8 μg/皿)。试验另设2个溶剂对照组(灭菌纯水和二甲基亚砜)、1个未处理对照即自发回变组、1个阳性对照组(阳性对照物敌克松、2-氨基芴和1,8-二羟基蒽醌溶剂为二甲基亚砜, 叠氮钠和环磷酰胺溶剂为灭菌纯水)。
采用平板掺入法进行试验, 每个测试点各设3个平行样。在2 mL顶层培养基中依次加入0.1 mL试验菌株增菌液, 0.1 mL受试物溶液(或溶剂、阳性对照物, 未处理对照组不加), 需要代谢活化时另加入S9混合液0.5 mL, 混匀后倒入底层培养基平板上, 培养基凝固后倒置于37 ℃培养箱中孵育48 h, 计数每皿中的回变菌落数。重复该试验1次, 除剂量间距不同外(5000.0、1250.0、312.5、78.1、19.5 μg/皿), 试验方法同上。
1.4.6 小鼠精母细胞染色体畸变试验
选用SPF级健康雄性ICR小鼠25只, 初始体重29~35 g。小鼠按体重随机分入3个样品剂量组[5000、2500、1250 mg/(kg·bw)]、1个溶剂对照组(纯水)和1个阳性对照组[丝裂霉素C, 2 mg/(kg·bw)], 每组5只。样品各剂量组、溶剂对照组动物均连续灌胃5 d, 每天1次, 每次灌胃容量均为20 mL/(kg·bw); 阳性对照组动物仅于试验第1 d腹腔注射1次, 注射量为10 mL/(kg·bw)。试验第14 d, 各组动物均于处死前4 h腹腔注射秋水仙素[5 mg/(kg·bw)], 注射量为10 mL/(kg·bw)。颈椎脱臼法处死小鼠, 取两侧睾丸, 去净脂肪, 撕开被膜, 分离曲细精管, 1%柠檬酸三钠低渗处理, 甲醇:冰乙酸(V:V=3:1)固定液固定两次, 800 r/min离心5 min后使用60%冰乙酸软化, 1000 r/min离心10 min后进行滴片, Giemsa液染色, 阅片。
1.5 数据处理
本研究中试验数据以平均数±标准偏差表示, 利用IBM SPSS Statistics 23.0 standard 网络版进行统计分析, 采用卡方检验对微核试验及染色体畸变试验中计数资料进行组间差异比较, P<0.05表示有统计学意义, P<0.01表示具有显著性差异。
2 结果与分析
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2.1 小鼠急性经口毒性试验结果
γ-PGA对小鼠体重的影响及急性经口毒性观察情况分别见表1、表2。经口给予小鼠γ-PGA后, 未见明显中毒表现, 对其体重增长也无明显影响, 整个观察期内未发生动物死亡。至观察期结束时, 对小鼠大体解剖检查同样未见明显异常。本次试验中, γ-PGA对雌雄小鼠MTD值均大于10000 mg/(kg·bw), 其半数致死剂量(median lethal dose, LD50)大于该数值, 根据急性经口毒性分级标准, γ-PGA属实际无毒级。
2.2 大鼠急性经口毒性试验结果
γ-PGA对大鼠体重的影响及急性经口毒性观察情况分别见表3、表4。大鼠灌胃约3 h后出现腹泻、粪便不成形等中毒表现, 约1 d后症状相继缓解, 整个观察期内未出现大鼠死亡的情况, 对其体重增长也无明显影响。至观察期结束时, 对大鼠大体解剖检查同样未见明显异常。本研究中, γ-PGA对雌雄大鼠急性经口MTD值均大于10000 mg/(kg· bw), 其LD50大于该数值, 根据急性经口毒性分级标准, γ-PGA属实际无毒级。
2.3 小鼠骨髓细胞微核试验结果
PCE数与总红细胞(PCE+NCE)数的比值是评估化合物细胞毒性的重要指标。在本研究中, γ-PGA各剂量组PCE数与总红细胞数的比值与溶剂对照组相比均未呈现统计学差异(P>0.05), 提示骨髓红细胞增殖并无明显受抑[20], 即γ-PGA无细胞毒性, 因此对含微核的PCE观察无明显影响[21]。含微核PCE率是微核试验中评价化合物对哺乳动物体细胞染色体损伤程度的关键指标。研究结果显示, 阳性对照组含微核PCE率与溶剂对照组相比具有显著性差异(P<0.01), 而γ-PGA各剂量组的含微核PCE率与溶剂对照组相比, 差异均无统计学意义(P>0.05), 也无剂量-反应关系。因此, γ-PGA对小鼠骨髓红细胞无致微核作用, 结果见表5。
2.4 细菌回复突变试验结果
回变菌落数是细菌回复突变试验中评价化合物诱导基因突变活性的关键指标, 其数值变化可以反映化合物对测试菌株DNA的致突变潜能。在本研究中, 两次试验结果显示, 阳性对照组(加或不加S9)回变菌落数均高于相应自发回变组的2倍, 自发回变组和2个溶剂对照组(加或不加S9)回变菌落数均在本实验室历史正常值范围内。γ-PGA各剂量组背景菌苔生长良好, 提示无明显细菌毒性, 其回变菌落数均未超过相应自发回变组值的2倍, 且无剂量-反应关系, 说明在有无S9代谢活化条件下, γ-PGA对这5种菌株均未呈现诱变活性, 结果见表6、表7。
2.5 小鼠精母细胞染色体畸变试验结果
小鼠精母细胞染色体畸变率是评价化合物对雄性生殖细胞致突变能力的重要指标。本研究结果显示, γ-PGA各剂量组染色体畸变细胞率与溶剂对照组相比, 差异均无统计学意义(P>0.05), 也无剂量-反应关系, 阳性对照组与溶剂对照组比较, 染色体畸变细胞率、性染色体和常染色体单价体差异均具有统计学意义(P<0.01), 说明γ-PGA对小鼠精母细胞染色体畸变结果为阴性, 结果见表8。
3 讨论与结论
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γ-PGA在食品领域的应用日益广泛, 但在其安全性评价方面, 特别是2014版《食品安全国家标准》实施之后, 国内外现有的研究资料仍显得相对不足。鉴于此背景, 本研究深入研究了γ-PGA的急性毒性和遗传毒性。
在急性经口毒性试验中, γ-PGA对雌雄小鼠、大鼠的MTD值均大于10000 mg/(kg·bw), 表明在急性经口暴露条件下, γ-PGA对小鼠和大鼠毒性极低, 按照急性经口毒性分级标准, γ-PGA属于实际无毒级。本次研究中, 虽然大鼠在灌胃后出现了短暂的腹泻和粪便不成形等中毒表现, 但这些症状在1 d后即自行缓解, 短期内未对大鼠的体重增长和生存造成显著影响。
本研究采用细菌回复突变试验、小鼠骨髓细胞微核试验以及小鼠精母细胞染色体畸变试验的方法组合, 全面评估γ-PGA遗传毒性[22-25]。其中, 细菌回复突变试验是利用组氨酸缺陷型的鼠伤寒沙门氏菌, 检测受试物是否具有直接或间接致突变性[26-27]。在本研究中采用推荐的5000 μg/皿作为最高剂量, 首次试验剂量间距设计为5倍, 试验数据显示, 在加及不加S9代谢活化条件下, γ-PGA对标准测试菌株均未表现出明显的诱变活性。有文献指出[28], 过大的剂量间距设计可能会漏检一些弱致突变物, 因此在第二次试验中缩小剂量间距至4倍, 再次验证了该阴性结果。小鼠骨髓红细胞微核试验和小鼠精母细胞染色体畸变试验是评价外源化学物对染色体损伤的两种经典方法[29-33]。在本研究中, 参考前期急性经口毒性试验结果, 即无法得出具体LD50时, 根据最大灌胃剂量[最大使用质量浓度250 mg/mL和最大灌胃容量20 mL/(kg·bw)]设计了这两个试验的高剂量组, 即5000 mg/(kg·bw)。试验结果显示, γ-PGA在试验剂量下未引起小鼠骨髓嗜多染红细胞微核率的增加(P>0.05), 也未引起小鼠初级精母细胞染色体畸变率的上升(P>0.05)。以上这些检测结果共同支持了γ-PGA无遗传毒性的结论。
综上所述, 在本研究条件下, γ-PGA安全、无毒且不具有遗传毒性, 但为了更全面地评估其在食品领域的应用潜力, 仍需考虑其潜在的长期毒性风险, 当进一步研究其长期暴露下的安全性。
文献
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2025年第16卷第11期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241222001
  • 接收时间：2024-12-22
  • 首发时间：2025-07-14
  • 出版时间：2025-06-15
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  • 收稿日期：2024-12-22
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    江苏省疾病预防控制中心毒理与风险评估研究所, 南京 210009

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* 俞萍(1973—), 女, 硕士, 主任医师, 主要研究方向为食品和新食品原料毒理学安全性评价与风险评估。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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