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Article(id=1151437192074719860, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1151437189243089177, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241216001, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1734278400000, receivedDateStr=2024-12-16, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752453619258, onlineDateStr=2025-07-14, pubDate=1749916800000, pubDateStr=2025-06-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752453619258, onlineIssueDateStr=2025-07-14, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752453619258, creator=13701087609, updateTime=1752453619258, 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=228, endPage=233, ext={EN=ArticleExt(id=1151895325276467780, articleId=1151437192074719860, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Determination of bongkrekic acid content in Hanzhong Mianpi by ultra performance liquid chromatography-tandem mass spectrometry combined with QuEChERS purification, columnId=1151895321388347923, journalTitle=Journal of Food Safety & Quality, columnName=Food Analysis and Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method for rapid determination of bongkrekic acid content in Hanzhong Mianpi by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS purification. Methods Hanzhong Mianpi samples were extracted by acetonitrile-water (8:2, V:V) ultrasonic extraction, purified by QuEChERS, concentrated, constant volume, separated by Thermo (100 mm× 2.1 mm, 1.8 μm) chromatographic column, gradient eluted by acetonitrile and 0.1% formic acid aqueous solution, quantified by standard curve method with electric spray ion source (ESI), multiple reaction monitoring (MRM) mode. Results The linear relationship of bongkrekic acid was good within the range of 1-100 ng/mL, with a correlation coefficient (r2) greater than 0.999. At 3 spiked levels of 2, 10 and 50 μg/kg, the spiked recovery rate was 86.7%-90.0%, the relative standard deviation (RSD) was 2.3%-2.9%, the limit of detection was 0.5 μg/kg, and the limit of quantitation was 1.5 μg/kg. All actual samples did not detect any citric acid. Conclusion This method has simple pre-treatment, fast analysis, high recovery rate, and high sensitivity, which can provide reference for the quality control of bongkrekic acid in Hanzhong Mianpi.

, correspAuthors=Wen HUANG, 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=Jun LU, Wen HUANG, Chong-Yong LI, Hui-Jun GOU, Shan-Na XU), CN=ArticleExt(id=1151895334990476135, articleId=1151437192074719860, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=QuEChERS净化结合超高效液相色谱-串联质谱法快速检测汉中面皮中米酵菌酸含量, columnId=1151895321958773274, journalTitle=食品安全质量检测学报, columnName=食品分析与检测, runingTitle=null, highlight=null, articleAbstract=

目的 建立一种QuEChERS前处理净化结合超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速检测汉中面皮中米酵菌酸含量的方法。方法 面皮样品采用乙腈-水(8:2, V:V)超声提取, QuEChERS净化处理, 浓缩、定容, 选用Thermo (100 mm×2.1 mm, 1.8 μm)色谱柱分离, 以乙腈和0.1%甲酸水溶液进行梯度洗脱, 电喷雾离子源(electron spray ionization, ESI), 负离子多反应监测(multiple reaction monitoring, MRM)模式, 以标准曲线法定量。结果 米酵菌酸在1~100 ng/mL范围内线性关系良好, 相关系数(r2)大于0.999, 在2、10、50 μg/kg低中高3个加标水平下, 加标回收率为86.7%~90.0%, 相对标准偏差(relative standard deviation, RSD)为2.3%~2.9%, 检出限为0.5 μg/kg, 定量限为1.5 μg/kg。所有实际样品均未检出米酵菌酸。结论 该方法前处理简单、分析快速、回收率高、灵敏度高, 可为汉中面皮中米酵菌酸的质量控制提供参考。

, correspAuthors=黄文, authorNote=null, correspAuthorsNote=
* 黄文(1988—), 男, 工程师, 主要研究方向为食品药品检验检测、食品生产工艺研究。E-mail:
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卢军(1983—), 男, 高级工程师, 主要研究方向为食品药品质量安全控制、生产工艺研究与检验检测。E-mail:

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卢军(1983—), 男, 高级工程师, 主要研究方向为食品药品质量安全控制、生产工艺研究与检验检测。E-mail:

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卢军(1983—), 男, 高级工程师, 主要研究方向为食品药品质量安全控制、生产工艺研究与检验检测。E-mail:

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Journal of Food Safety & Quality, 2024, 15(3): 141-147., articleTitle=Preparation and identifeication of monoclonal antibodies of bongkrekic acid, refAbstract=null)], funds=[Fund(id=1167030694204158218, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, awardId=2022KY27, language=CN, fundingSource=陕西省市场监督管理局2022年度科技计划项目(2022KY27), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1167030691725324508, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, xref=null, ext=[AuthorCompanyExt(id=1167030691729518813, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, companyId=1167030691725324508, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Food and Drug Supervision Inspection and Testing Center of Hanzhong, Hanzhong 723000, China), AuthorCompanyExt(id=1167030691737907422, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, companyId=1167030691725324508, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=汉中市食品药品监督检验检测中心, 汉中 723000)])], figs=[ArticleFig(id=1167030693243662592, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=EN, label=Fig.1, caption=Effects of different extraction solvents on recovery rate, figureFileSmall=J0T9Rg8zoqwUv/ddvWC4jQ==, figureFileBig=BrmS+HhuqGBIfJHwYjckuQ==, tableContent=null), ArticleFig(id=1167030693327548673, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=CN, label=图1, caption=不同提取溶剂对回收率的影响, figureFileSmall=J0T9Rg8zoqwUv/ddvWC4jQ==, figureFileBig=BrmS+HhuqGBIfJHwYjckuQ==, tableContent=null), ArticleFig(id=1167030693449183490, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=EN, label=Fig.2, caption=Effects of different purification materials on recovery rate, figureFileSmall=tKZ8WFguK+xQGx0sEW/Y2Q==, figureFileBig=0DzEn843Oeu3ufyfZSPJ/Q==, tableContent=null), ArticleFig(id=1167030693533069571, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=CN, label=图2, caption=不同净化材料对回收率的影响, figureFileSmall=tKZ8WFguK+xQGx0sEW/Y2Q==, figureFileBig=0DzEn843Oeu3ufyfZSPJ/Q==, tableContent=null), ArticleFig(id=1167030693625344260, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=EN, label=Table 1, caption=

Gradient programs of mobile phase

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 0.1%甲酸水溶液/% 乙腈/%
0.0 80 20
2.0 30 70
4.0 30 70
5.0 80 20
6.0 80 20
), ArticleFig(id=1167030693780533509, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=CN, label=表1, caption=

流动相梯度洗脱程序

, figureFileSmall=null, figureFileBig=null, tableContent=
时间/min 0.1%甲酸水溶液/% 乙腈/%
0.0 80 20
2.0 30 70
4.0 30 70
5.0 80 20
6.0 80 20
), ArticleFig(id=1167030693918945542, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=EN, label=Table 2, caption=

Recoveries and precisions

, figureFileSmall=null, figureFileBig=null, tableContent=
标准物质名称 添加浓度/(μg/kg) 平均浓度/(μg/kg) 平均回收率/% RSDs/%
BA 2 1.734 86.7 2.6
10 8.910 89.1 2.9
50 45.000 90.0 2.3
), ArticleFig(id=1167030693969277191, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=CN, label=表2, caption=

回收率和精密度

, figureFileSmall=null, figureFileBig=null, tableContent=
标准物质名称 添加浓度/(μg/kg) 平均浓度/(μg/kg) 平均回收率/% RSDs/%
BA 2 1.734 86.7 2.6
10 8.910 89.1 2.9
50 45.000 90.0 2.3
), ArticleFig(id=1167030694019608840, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=EN, label=Table 3, caption=

Comparison of the established method with the national standard method and reference literature

, figureFileSmall=null, figureFileBig=null, tableContent=
项目 方法
前处理步骤 使用试剂 实验时间/h
液液萃取、QuEChERS净化 3种, 无酸碱试剂使用 1.0~1.5 本研究
提取、浸泡1 h、重复提取, 固相萃取净化 6种, 有酸碱试剂使用 2.0~2.5 GB 5009.189—2023
液液萃取、QuEChERS净化 4种, 无酸碱试剂使用 1.0~1.5 [23]
提取、浸泡1 h、混合阴离子交换柱净化 5种, 有酸碱试剂使用 2.0~2.5 [24]
液液萃取、混合强阴离子交换柱净化 7种, 有酸碱试剂使用 1.0~1.5 [25]
提取、浸泡1 h 1种, 有酸碱试剂使用 2.0~2.5 [28]
), ArticleFig(id=1167030694082523401, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1151437192074719860, language=CN, label=表3, caption=

所建方法与国标方法及参考文献对比

, figureFileSmall=null, figureFileBig=null, tableContent=
项目 方法
前处理步骤 使用试剂 实验时间/h
液液萃取、QuEChERS净化 3种, 无酸碱试剂使用 1.0~1.5 本研究
提取、浸泡1 h、重复提取, 固相萃取净化 6种, 有酸碱试剂使用 2.0~2.5 GB 5009.189—2023
液液萃取、QuEChERS净化 4种, 无酸碱试剂使用 1.0~1.5 [23]
提取、浸泡1 h、混合阴离子交换柱净化 5种, 有酸碱试剂使用 2.0~2.5 [24]
液液萃取、混合强阴离子交换柱净化 7种, 有酸碱试剂使用 1.0~1.5 [25]
提取、浸泡1 h 1种, 有酸碱试剂使用 2.0~2.5 [28]
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QuEChERS净化结合超高效液相色谱-串联质谱法快速检测汉中面皮中米酵菌酸含量
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卢军 , 黄文 * , 李崇勇 , 缑慧君 , 徐姗娜
食品安全质量检测学报 | 食品分析与检测 2025,16(11): 228-233
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食品安全质量检测学报 | 食品分析与检测 2025, 16(11): 228-233
QuEChERS净化结合超高效液相色谱-串联质谱法快速检测汉中面皮中米酵菌酸含量
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卢军 , 黄文* , 李崇勇, 缑慧君, 徐姗娜
作者信息
  • 汉中市食品药品监督检验检测中心, 汉中 723000

    • 卢军(1983—), 男, 高级工程师, 主要研究方向为食品药品质量安全控制、生产工艺研究与检验检测。E-mail:

通讯作者:

* 黄文(1988—), 男, 工程师, 主要研究方向为食品药品检验检测、食品生产工艺研究。E-mail:
Determination of bongkrekic acid content in Hanzhong Mianpi by ultra performance liquid chromatography-tandem mass spectrometry combined with QuEChERS purification
Jun LU , Wen HUANG* , Chong-Yong LI, Hui-Jun GOU, Shan-Na XU
Affiliations
  • Food and Drug Supervision Inspection and Testing Center of Hanzhong, Hanzhong 723000, China
出版时间: 2025-06-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241216001
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摘要
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目的 建立一种QuEChERS前处理净化结合超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速检测汉中面皮中米酵菌酸含量的方法。方法 面皮样品采用乙腈-水(8:2, V:V)超声提取, QuEChERS净化处理, 浓缩、定容, 选用Thermo (100 mm×2.1 mm, 1.8 μm)色谱柱分离, 以乙腈和0.1%甲酸水溶液进行梯度洗脱, 电喷雾离子源(electron spray ionization, ESI), 负离子多反应监测(multiple reaction monitoring, MRM)模式, 以标准曲线法定量。结果 米酵菌酸在1~100 ng/mL范围内线性关系良好, 相关系数(r2)大于0.999, 在2、10、50 μg/kg低中高3个加标水平下, 加标回收率为86.7%~90.0%, 相对标准偏差(relative standard deviation, RSD)为2.3%~2.9%, 检出限为0.5 μg/kg, 定量限为1.5 μg/kg。所有实际样品均未检出米酵菌酸。结论 该方法前处理简单、分析快速、回收率高、灵敏度高, 可为汉中面皮中米酵菌酸的质量控制提供参考。

超高效液相色谱-串联质谱法  /  QuEChERS净化  /  米酵菌酸  /  汉中面皮

Objective To establish a method for rapid determination of bongkrekic acid content in Hanzhong Mianpi by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) combined with QuEChERS purification. Methods Hanzhong Mianpi samples were extracted by acetonitrile-water (8:2, V:V) ultrasonic extraction, purified by QuEChERS, concentrated, constant volume, separated by Thermo (100 mm× 2.1 mm, 1.8 μm) chromatographic column, gradient eluted by acetonitrile and 0.1% formic acid aqueous solution, quantified by standard curve method with electric spray ion source (ESI), multiple reaction monitoring (MRM) mode. Results The linear relationship of bongkrekic acid was good within the range of 1-100 ng/mL, with a correlation coefficient (r2) greater than 0.999. At 3 spiked levels of 2, 10 and 50 μg/kg, the spiked recovery rate was 86.7%-90.0%, the relative standard deviation (RSD) was 2.3%-2.9%, the limit of detection was 0.5 μg/kg, and the limit of quantitation was 1.5 μg/kg. All actual samples did not detect any citric acid. Conclusion This method has simple pre-treatment, fast analysis, high recovery rate, and high sensitivity, which can provide reference for the quality control of bongkrekic acid in Hanzhong Mianpi.

ultra performance liquid chromatography-tandem mass spectrometry  /  QuEChERS purification  /  bongkrekic acid  /  Hanzhong Mianpi
卢军, 黄文, 李崇勇, 缑慧君, 徐姗娜. QuEChERS净化结合超高效液相色谱-串联质谱法快速检测汉中面皮中米酵菌酸含量. 食品安全质量检测学报, 2025 , 16 (11) : 228 -233 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241216001
Jun LU, Wen HUANG, Chong-Yong LI, Hui-Jun GOU, Shan-Na XU. Determination of bongkrekic acid content in Hanzhong Mianpi by ultra performance liquid chromatography-tandem mass spectrometry combined with QuEChERS purification[J]. Journal of Food Safety & Quality, 2025 , 16 (11) : 228 -233 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241216001
正文
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0 引言
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米酵菌酸(bongkrekic acid, BA)是一种椰毒假单胞菌酵米面亚种产生的一种作用于线粒体的生物毒素[1-5], 能够阻止线粒体中的氧化磷酸化过程, 最终导致中毒者肝脏、肾脏、大脑等多处器官功能衰竭[6-7]、凝血功能异常、代谢性酸中毒, 具有极高的致死率[8-10]。在变质的发酵玉米面制品和变质淀粉类制品中, BA通过椰毒假单胞菌米面亚种的繁殖而不断累积。BA耐热, 一般烹调方法不能破坏其毒性。摄入后会引起食物中毒, 轻者表现为头晕、恶心、呕吐、全身无力等症状[11-12], 重者可出现黄疸、肝肿大、皮下出血、呕血、血尿、意识不清、抽搐等[13], 甚至死亡[14]。
汉中面皮是以大米为原料, 经过浸泡、磨浆(添加或不添加食用淀粉)、蒸熟、切条, 再配蔬菜和作料的一种地方特色美食, 冷热形态下均可食用, 具有白薄、光滑、细嫩、柔韧、鲜香等特点, 口感软糯, 香辣味美, 回味悠长, 老少皆宜[15-17]。汉中面皮的加工工艺与湿米粉制品基本相同, 其本质也属于米粉制品, 加上汉中本身处于盆地, 夏季气温高、潮湿, 生产、运输、经营、消费各环节均存在较大的BA污染风险。而BA的热稳定性, 使其在高温烹饪环节仍然保持高毒性。由于汉中面皮在人们生活中的重要性和BA污染的高风险性, 因此建立一种高效、准确、简便地检测汉中面皮中的BA的定性和定量方法势在必行。
目前针对BA的检测方法[18-19]主要有高效液相色谱法[20-22]、液相色谱-串联质谱法[23-28]和快速检测法[29-32]等。本研究在参考GB 5009.189—2023《食品安全标准 食品中米酵菌酸的测定》及目前BA的仪器检测、样品制备方法, 改良QuEChERS净化技术, 建立了超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry, UPLC-MS/MS)快速检测汉中面皮中BA含量的方法, 使其为汉中面皮中BA的快速检测提供技术参考和数据依据, 以期该方法能促进汉中面皮的生产、流通和消费环节中安全监管和质量控制水平的提升, 对促进汉中面皮产业的发展具有非常重要的意义。
1 材料与方法
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1.1 材料与试剂
汉中面皮: 市售, 样品为超市、农贸市场、菜市场随机购买的样品。
阴性样品: 自制, 按照汉中面皮制作工艺使用新制米浆制作并保存在2~8 ℃冷柜中, 经GB 5009.189—2023第二法 液相色谱-质谱/质谱法测定, 未检出BA。
BA标准品(C28H38O7, CAS号: 11076-19-0, 纯度≥95%)(坛墨质检科技股份有限公司); 甲醇、乙腈、甲酸(色谱纯)、甲酸铵(优级纯)(美国赛默飞世尔科技公司)。
1.2 仪器与设备
Thermo TSQ Quantis超高效液相色谱-串联质谱仪(美国赛默飞世尔科技公司); BS224S电子分析天平(精度0.1 mg, 德国赛多利斯公司); H1850高速冷冻离心机(湖南湘仪实验仪器有限公司); VORTEX-5漩涡振荡器(海门市其林贝尔仪器制造有限公司); KQ-500DE超声波清洗器(昆山市超声仪器有限公司); MFV-24氮吹仪(得泰仪器科技有限公司); Milli Q超纯水系统(美国Millipore公司)。
1.3 实验方法
1.3.1 前处理方法
(1)提取。面皮样品粉碎后, 精密称取2.5000 g(精确到0.1 mg)于50.0 mL塑料离心管中, 加入10.00 mL乙腈-水溶液(8:2, V:V), 充分涡旋混匀, 静置10 min, 剧烈振荡2 min, 超声提取20 min, 离心5 min (4000 r/min), 待净化。
(2)净化。将提取后上清液全部转移到QuEChERS dSPE EMR-Lipid离心管中, 摇匀, 涡旋振荡2 min, 离心5 min (4000 r/min), 离心管中上清液转移至已装有NaCl和无水MgSO4的净化管中, 充分混匀, 涡旋振荡2 min, 离心5 min (4000 r/min), 收集净化管中上清液, 于40 ℃水浴下氮气吹干, 准确加入1.0 mL乙腈-水溶液(8:2, V:V)复溶, 经0.22 µm微孔滤膜过滤, 供UPLC-MS/MS检测备用。
1.3.2 色谱条件
色谱柱为Thermo (100 mm×2.1 mm, 1.8 μm); 流动相分别为: 0.1%甲酸水溶、乙腈; 流速: 0.3 mL/min; 柱温: 40 ℃; 进样量: 5 µL; 流动相梯度洗脱程序见表1。
1.3.3 质谱条件
质谱参数: 离子化方式为电喷雾离子源(electron spray ionization, ESI); 负离子模式; 多反应监测(multiple reaction monitoring, MRM); 喷雾电压为3000 V; 离子传输管温度为320 ℃; 离子选择参数: 定量离子(m/z 485.300→441.333), 定性离子(m/z 485.300→397.083)。
1.4 数据处理
通过TraceFinder 4.1软件(美国赛默飞世尔科技公司)进行数据采集和预处理、生成标准曲线, 样品做6次平行实验, 并使用外标法定量分析; 采用WPS office 12.1.0.19302软件制表及绘制柱状图, 进行数据统计与比较。
2 结果与分析
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2.1 质谱条件优化
将仪器调节至优化模式, 针对BA选择ESI源, 分别进行正离子模式和负离子模式的全扫描, 比较得出BA在ESI负离子模式下响应值较好, 因此使用ESI负离子模式进行分析。选用母离子m/z 485.300进行优化, 在优化的二级子离子中m/z 441.333响应值最高, m/z 397.083响应值第二高, 故选择作为m/z 485.300/441.333定量离子对, 选择m/z 485.300/397.083作为定性离子对。
2.2 色谱条件优化
本研究对比了甲醇-水、甲醇-0.1%甲酸水、甲醇-2 mmol/L甲酸铵、乙腈-水、乙腈-0.1%甲酸水、乙腈-2 mmol/L甲酸铵作为流动相时, 测试BA在流动相中的电离强度, 结果显示选取乙腈-0.1%甲酸水作为流动相时电离强度最高。同时比较了甲醇和乙腈作为流动相的有机相, 使用乙腈峰型更好且柱压更低。综上可得, 理想流动相应选择乙腈-0.1%甲酸水, 同时将洗脱程序进行了优化, 使得BA得到了更好的分离, 峰型更好, 出峰时间更快, 最终得出最优梯度洗脱程序(见表1)。
2.3 前处理方法优化
2.3.1 提取溶剂优化
以阴性样品为基质, 选取乙腈、乙腈-水(8:2, V:V)、乙腈-0.1%甲酸水、甲醇、甲醇-水(8:2, V:V)作为提取溶剂进行测试。分别向阴性样品加入BA标准工作液, 将上述5种提取溶剂各10 mL分别加入样品中, 充分混匀, 静置10 min, 涡旋振荡2 min, 超声提取20 min, 4000 r/min离心5 min, 经0.22 µm尼龙微孔滤膜过滤, 测定BA含量, 计算出回收率, 结果见图1, 结果显示以乙腈-水(8:2, V:V)为提取溶液回收率最高, 因此提取溶剂选择乙腈-水(8:2, V:V)。
2.3.2 净化材料的优化
本研究对比了C18、乙二胺-N-丙基硅烷(primary secondary amine, PSA)、C18+PSA、dSPE EMR-Lipid不同净化材料对BA加标回收率的影响。以阴性样品为基质, 分别加入BA标准工作液, 经乙腈-水(8:2, V:V)提取后, 分别使用上述4种净化材料进行净化, 结果显示(图2), C18、PSA、C18+PSA这3类净化材料均对BA有较强的吸附, 极大影响了回收率。使用dSPE EMR-Lipid净化材料时既能较好地去除杂质, 也能尽可能低地减少对BA的吸附, 从而提高回收率。因此净化材料选择dSPE EMR-Lipid。
2.4 方法学评价
2.4.1 线性关系、检出限和定量限
将BA标准物质分别配制成质量浓度为1、2、5、10、20、50、100 ng/mL的系列标准溶液, 建立标准品工作曲线(BA质量浓度为横坐标(X, ng/mL), BA相应峰面积(Y)为纵坐标)。将标准溶液稀释后加入空白基质中, 进行1.3.1前处理方法处理后上机测定, 以3倍信噪比(S/N)和10倍信噪比(S/N)分别作为检出限和方法定量限。BA的线性方程为Y=3.853e3X-91.9 (r2=0.9996), 在1~100 ng/mL范围内线性关系良好, 检出限和定量限分别为0.5 µg/kg、1.5 µg/kg, 均满足GB 5009.189—2023的要求。
2.4.2 回收率和精密度
精密称取阴性样品2.5000 g, 分别量取对应低、中、高3种浓度标准溶液于样品中, 每种浓度平行处理6个样品, 样品按照1.3.1前处理方法进行处理, 上机测定, 根据添加浓度计算出回收率, 结果见表2, 由表2可见BA回收率在86.7%~90.0%之间, 符合GB/T 27417—2017《合格评定 化学分析方法确认和验证指南》中方法回收率偏差要求, RSDs在2.3%~2.9%之间。
2.5 实际样品测定
在生产环节和流通环节购买15批汉中面皮样品, 按1.3.1项下制备供试品溶液, 按照1.3.2项下色谱条件和1.3.3项下质谱条件进行测定, 结果显示15批汉中面皮样
品均未检出BA。
2.6 方法对比
所建方法与GB 5009.189—2023法相比(表3), 该方法在前处理环节处理更便捷和高效, 仪器分析时间也短于国家标准方法。
目前, 针对BA测定的现行国家标准为GB 5009.189—2023, 其中第二法液相色谱-质谱/质谱法仅仅提高了检测的准确度和灵敏度, 但是在前处理过程中仍存在耗时且酸碱试剂使用量较大的问题。梁明等[23]、曾雪芳等[24]、周鹏[25]、于浩洋等[28]均采用UPLC-MS/MS, 提高了检测的效率, 但在提取效率上各有优劣(见表3), 本研究兼顾提取和检测两方面, 旨在简化提取步骤、减少试剂用量、缩短实验时间上提高检测效率。
3 结论
收起
本研究通过改进前处理方法, 建立了UPLC-MS/MS快速检测汉中面皮中BA的含量。该方法在GB 5009.189—2023的基础上对前处理方法、色谱条件和质谱条件进行了优化, BA在1~100 ng/mL范围内线性关系良好, 相关系数(r2)大于0.999, 方法检出限、定量限均高于国家标准中对于食品中BA的测定, 在低、中、高3个浓度水平加标回收率范围为86.7%~90.0%, RSDs在2.3%~2.9%, 并且大大缩短了实验时间, 由此说明该方法可以为制定汉中面皮食品安全地方标准提供可靠的数据参考与技术支持。
基金
收起
  • 陕西省市场监督管理局2022年度科技计划项目(2022KY27)
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241216001
  • 接收时间：2024-12-16
  • 首发时间：2025-07-14
  • 出版时间：2025-06-15
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  • 收稿日期：2024-12-16
基金
陕西省市场监督管理局2022年度科技计划项目(2022KY27)
作者信息
    汉中市食品药品监督检验检测中心, 汉中 723000

通讯作者:

* 黄文(1988—), 男, 工程师, 主要研究方向为食品药品检验检测、食品生产工艺研究。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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