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Article(id=1240722577677677228, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240722566957027366, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202505145, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1746979200000, receivedDateStr=2025-05-12, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773740914029, onlineDateStr=2026-03-17, pubDate=1760025600000, pubDateStr=2025-10-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773740914029, onlineIssueDateStr=2026-03-17, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773740914029, creator=13701087609, updateTime=1773740914029, updator=13701087609, issue=Issue{id=1240722566957027366, tenantId=1146029695717560320, journalId=1227665162245664772, year='2025', volume='52', issue='19', pageStart='3457', pageEnd='3648', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=0, createTime=1773740911472, creator=13701087609, updateTime=1773740981732, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240722861736906836, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240722566957027366, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240722861736906837, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240722566957027366, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=3615, endPage=3619, ext={EN=ArticleExt(id=1240722578097107662, articleId=1240722577677677228, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Determination of biomarkers of propylene glycol monomethyl ether in urine by paper spray mass spectrometry, columnId=1228016572065837304, journalTitle=Modern Preventive Medicine, columnName=Experimental Technology and Applications, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a paper spray mass spectrometry method for the determination of 1-methoxy-2-propanol and 2-methoxypropionic acid as biomarkers of propylene glycol monomethyl ether in urine. Methods The urine sample was dropped onto the chromatographic paper, and a high voltage of 4 kV was applied to the back of the paper. After dissolution in the spray solvent, the sample formed an electrospray at the paper tip under the effect of voltage, which then entered the mass spectrometer for detection. Results The linear ranges of 1-methoxy-2-propanol and 2-methoxypropionic acid were 0.036-46.15 μg/mL and 0.029-54.25 μg/mL, respectively, with correlation coefficients ranging from 0.999 6 to 0.999 9. The detection limit (LOD) was between 0.008 8 and 0.011 μg/mL, and the quantification limit(LOQ) was between 0.029 and 0.036 μg/mL. The sample recovery rate ranged from 90.8% to 105.0%, and RSDs ranged from 3.3% to 9.7%. Conclusion This method requires no sample processing, features simplicity, rapidity and high sensitivity, making it suitable for the rapid screening of propylene glycol monomethyl ether in urine.

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目的 建立尿液中丙二醇单甲醚的生物标志物1-甲氧基-2-丙醇和2-甲氧基丙酸的纸喷雾质谱测定法。方法 将尿液样品滴加于色谱纸，在纸片后端施加4 kV高压，样品溶于喷雾溶剂后在电压作用下于纸尖端形成电喷雾，继而进入质谱仪检测。结果 1-甲氧基-2-丙醇和2-甲氧基丙酸的线性范围分别为0.036～46.15 μg/ml、0.029～54.25 μg/ml，相关系数在0.999 6～0.999 9之间，检出限在0.008 8～0.011 μg/ml之间，定量限在0.029～0.036 μg/ml之间，样品加标回收率为90.8%～105.0%，精密度的相对标准偏差为3.3%～9.7%。结论 本法无需样品处理，简便快速、灵敏，适用于尿液中丙二醇单甲醚的快速筛查。

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刘保峰，E-mail:
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刘蒙蒙（1990—），女，硕士，助理研究员，研究方向：职业病危害因素监测、职业中毒预防与控制

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刘蒙蒙（1990—），女，硕士，助理研究员，研究方向：职业病危害因素监测、职业中毒预防与控制

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刘蒙蒙（1990—），女，硕士，助理研究员，研究方向：职业病危害因素监测、职业中毒预防与控制

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Analytical Chemistry, 2024, 96(42): 16706–16714., articleTitle=Paper spray ionization mass spectrometry coupled with Paper-Based Three-Dimensional tumor model for rapid metabolic gradient profiling, refAbstract=null)], funds=[Fund(id=1240933509624296238, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, awardId=TJWJ2023QN087, language=CN, fundingSource=天津市卫生健康科研项目(TJWJ2023QN087), fundOrder=null, country=null), Fund(id=1240933509741736760, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, awardId=TJYXZDXK-3-020B, language=CN, fundingSource=天津市医学重点建设学科(TJYXZDXK-3-020B), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1240933505148973585, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, xref=1., ext=[AuthorCompanyExt(id=1240933505157362193, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, companyId=1240933505148973585, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Department of Occupational Health, Tianjin Center for Disease Control and Prevention, Tianjin 300011, China), AuthorCompanyExt(id=1240933505165750804, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, companyId=1240933505148973585, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.天津市疾病预防控制中心职业健康所，天津 300011)]), AuthorCompany(id=1240933505278997020, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, xref=2., ext=[AuthorCompanyExt(id=1240933505283191327, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, companyId=1240933505278997020, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.天津医科大学公共卫生学院)])], figs=[ArticleFig(id=1240933508198232812, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Figure 1, caption=PS-MS process schematic and device diagram, figureFileSmall=fN9NMYn+vXU35usBJJLYBg==, figureFileBig=VZCDRQ/EV1itKyUu2O6O1Q==, tableContent=null), ArticleFig(id=1240933508294701810, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=图1, caption=纸喷雾质谱技术过程示意图和装置图

注：图A为过程示意图；图B为装置图。

, figureFileSmall=fN9NMYn+vXU35usBJJLYBg==, figureFileBig=VZCDRQ/EV1itKyUu2O6O1Q==, tableContent=null), ArticleFig(id=1240933508416336630, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Figure 2, caption=Primary mass spectra of α-PGME and 2-MPA, figureFileSmall=BiJpWaBiwusnRvnMphtsiA==, figureFileBig=qsBx3UIg8HxFAykXd0nIog==, tableContent=null), ArticleFig(id=1240933508525388536, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=图2, caption=α-PGME和2-MPA的一级质谱图

注：图A为α-PGME；图B为2-MPA。

, figureFileSmall=BiJpWaBiwusnRvnMphtsiA==, figureFileBig=qsBx3UIg8HxFAykXd0nIog==, tableContent=null), ArticleFig(id=1240933508651217662, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Figure 3, caption=Optimization of paper spray ionization conditions, figureFileSmall=/xoosvbpcZvzPJOKe887xQ==, figureFileBig=ys2UYBIgwtyx0h9jwN7F7g==, tableContent=null), ArticleFig(id=1240933508743492353, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=图3, caption=纸喷雾电离条件的优化

注：A为纸基质尖端到质谱进样口的距离优化图；B为施加电压强度优化图；C为喷雾溶剂种类优化图；D为喷雾溶剂用量优化图。

, figureFileSmall=/xoosvbpcZvzPJOKe887xQ==, figureFileBig=ys2UYBIgwtyx0h9jwN7F7g==, tableContent=null), ArticleFig(id=1240933508835767046, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Table 1, caption=

Mass spectrometric parameters of α-PGME and 2-MPA

, figureFileSmall=null, figureFileBig=null, tableContent=
物质化学式极性前体离子（m/z）产物离子（m/z）碰撞能量（eV）
α-PGMEC4H10O2+73.0841/43/45*35
2-MPAC4H8O3—103.3371*35
), ArticleFig(id=1240933508944818955, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=表1, caption=

α-PGME和2-MPA的质谱参数

, figureFileSmall=null, figureFileBig=null, tableContent=
物质化学式极性前体离子（m/z）产物离子（m/z）碰撞能量（eV）
α-PGMEC4H10O2+73.0841/43/45*35
2-MPAC4H8O3—103.3371*35
), ArticleFig(id=1240933509070648079, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Table 2, caption=

The regression equations, correlation coefficients, detection limits, and quantification limits of the method

, figureFileSmall=null, figureFileBig=null, tableContent=
被测物回归方程相关系数检出限（μg/ml）定量下限（μg/ml）
α-PGMEy = 14 094x + 1 902.80.999 60.011 00.036
2-MPAy = 21 430x + 996.90.999 90.008 80.029
), ArticleFig(id=1240933509175505685, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=表2, caption=

方法的回归方程、相关系数、检出限和定量限

, figureFileSmall=null, figureFileBig=null, tableContent=
被测物回归方程相关系数检出限（μg/ml）定量下限（μg/ml）
α-PGMEy = 14 094x + 1 902.80.999 60.011 00.036
2-MPAy = 21 430x + 996.90.999 90.008 80.029
), ArticleFig(id=1240933509288751899, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=EN, label=Table 3, caption=

The recovery rates and precisions of the method（n=6）

, figureFileSmall=null, figureFileBig=null, tableContent=
被测物本底值（μg/ml）加标量（μg/ml）测定量（μg/ml）回收率（%）RSD（%）
α-PGMEND46.150 047.580 0103.15.9
ND4.615 04.633 0100.43.5
ND0.184 60.173 293.89.7
2-MPAND54.250 050.340 092.88.0
ND5.425 05.696 0105.06.8
ND0.217 00.197 090.83.3
), ArticleFig(id=1240933509389415200, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240722577677677228, language=CN, label=表3, caption=

方法的加标回收率和精密度（n=6）

, figureFileSmall=null, figureFileBig=null, tableContent=
被测物本底值（μg/ml）加标量（μg/ml）测定量（μg/ml）回收率（%）RSD（%）
α-PGMEND46.150 047.580 0103.15.9
ND4.615 04.633 0100.43.5
ND0.184 60.173 293.89.7
2-MPAND54.250 050.340 092.88.0
ND5.425 05.696 0105.06.8
ND0.217 00.197 090.83.3
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纸喷雾质谱法测定尿液中丙二醇单甲醚的生物标志物
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刘蒙蒙 1 , 潘俊宇 1 , 吕勃熠 1 , 刘保峰 1 , 曾强 1, 2
现代预防医学 | 实验技术及其应用 2025,52(19): 3615-3619
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现代预防医学 | 实验技术及其应用 2025, 52(19): 3615-3619
纸喷雾质谱法测定尿液中丙二醇单甲醚的生物标志物
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刘蒙蒙1, 潘俊宇1, 吕勃熠1, 刘保峰1 , 曾强1, 2
作者信息
  • 1.天津市疾病预防控制中心职业健康所，天津 300011
  • 2.天津医科大学公共卫生学院

    • 刘蒙蒙（1990—），女，硕士，助理研究员，研究方向：职业病危害因素监测、职业中毒预防与控制

通讯作者:

刘保峰，E-mail:
Determination of biomarkers of propylene glycol monomethyl ether in urine by paper spray mass spectrometry
Meng-meng LIU1, Jun-yu PAN1, Bo-yi LV1, Bao-feng LIU1 , Qiang ZENG1, 2
Affiliations
  • Department of Occupational Health, Tianjin Center for Disease Control and Prevention, Tianjin 300011, China
出版时间: 2025-10-10 doi: 10.20043/j.cnki.MPM.202505145
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摘要
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目的 建立尿液中丙二醇单甲醚的生物标志物1-甲氧基-2-丙醇和2-甲氧基丙酸的纸喷雾质谱测定法。方法 将尿液样品滴加于色谱纸，在纸片后端施加4 kV高压，样品溶于喷雾溶剂后在电压作用下于纸尖端形成电喷雾，继而进入质谱仪检测。结果 1-甲氧基-2-丙醇和2-甲氧基丙酸的线性范围分别为0.036～46.15 μg/ml、0.029～54.25 μg/ml，相关系数在0.999 6～0.999 9之间，检出限在0.008 8～0.011 μg/ml之间，定量限在0.029～0.036 μg/ml之间，样品加标回收率为90.8%～105.0%，精密度的相对标准偏差为3.3%～9.7%。结论 本法无需样品处理，简便快速、灵敏，适用于尿液中丙二醇单甲醚的快速筛查。

尿液  /  纸喷雾质谱法  /  丙二醇单甲醚  /  代谢产物

Objective To establish a paper spray mass spectrometry method for the determination of 1-methoxy-2-propanol and 2-methoxypropionic acid as biomarkers of propylene glycol monomethyl ether in urine. Methods The urine sample was dropped onto the chromatographic paper, and a high voltage of 4 kV was applied to the back of the paper. After dissolution in the spray solvent, the sample formed an electrospray at the paper tip under the effect of voltage, which then entered the mass spectrometer for detection. Results The linear ranges of 1-methoxy-2-propanol and 2-methoxypropionic acid were 0.036-46.15 μg/mL and 0.029-54.25 μg/mL, respectively, with correlation coefficients ranging from 0.999 6 to 0.999 9. The detection limit (LOD) was between 0.008 8 and 0.011 μg/mL, and the quantification limit(LOQ) was between 0.029 and 0.036 μg/mL. The sample recovery rate ranged from 90.8% to 105.0%, and RSDs ranged from 3.3% to 9.7%. Conclusion This method requires no sample processing, features simplicity, rapidity and high sensitivity, making it suitable for the rapid screening of propylene glycol monomethyl ether in urine.

Urine  /  Paper spray mass spectrometry  /  Propylene glycol monomethyl ether  /  Metabolites
刘蒙蒙, 潘俊宇, 吕勃熠, 刘保峰, 曾强. 纸喷雾质谱法测定尿液中丙二醇单甲醚的生物标志物. 现代预防医学, 2025 , 52 (19) : 3615 -3619 . DOI: 10.20043/j.cnki.MPM.202505145
Meng-meng LIU, Jun-yu PAN, Bo-yi LV, Bao-feng LIU, Qiang ZENG. Determination of biomarkers of propylene glycol monomethyl ether in urine by paper spray mass spectrometry[J]. Modern Preventive Medicine, 2025 , 52 (19) : 3615 -3619 . DOI: 10.20043/j.cnki.MPM.202505145
正文
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丙二醇单甲醚（propylene glycol monomethyl ether，PGME）因兼具良好的水溶性、脂溶性和适中挥发性，被广泛应用于化妆品、涂料、油墨、皮革处理及工业清洗等领域[1]。毒理学研究表明，急性接触PGME可引起眼、上呼吸道和皮肤刺激，以及头痛、恶心、眩晕、嗜睡等中枢抑制症状[2-3]。本课题组[4]检测天津市10家汽修企业的48份原辅料样本发现，PGME检出率为25%。其广泛使用使职业人群暴露风险显著高于普通人群，职业健康风险亟待关注。
工业用PGME主要以两种异构体存在，主要形式为1-甲氧基-2-丙醇（α-PGME），次要形式为 2-甲氧基-1-丙醇（β-PGME）。α-PGME在体内经微粒体混合功能氧化酶（MFO）系统，进行O-去甲基化代谢，大部分以二氧化碳呼出，少部分以原形从尿中排出。而β-PGME经醇脱氢酶和醛脱氢酶代谢为2-甲氧基丙酸（2-MPA）[3]。为监测职业人群PGME暴露水平，课题组前期建立了工作场所空气中PGME的气相色谱测定法[5]。生物监测作为空气监测的补充，能直观反映劳动者接触水平，对保护劳动者健康具有重要意义。
目前尿液中PGME生物标志物的检测以气相色谱法为主[6]，样品前处理技术主要采用顶空固相微萃取法和液液萃取法。但这些方法存在操作繁琐、使用有毒有机溶剂、耗时较长等不足，不仅对操作人员健康有潜在风险，也不利于快速检测，且目前尚无同时测定PGME两种生物标志物的方法。
纸喷雾质谱技术的原理是将生物样品加载于纸基质表面，通过高压电驱动溶剂溶解、迁移并电离目标化合物，在纸尖端形成电喷雾后由质谱实时在线监测[7]。该技术样品用量少，可分析血液和尿液等生物样本中多种目标物，且分辨率高。纸喷雾质谱技术高效快捷，单个样品平均1～2 min内即可完成分析[8]。如1 min内可完成血液中的敌草快、百草枯、草甘膦等物质的检测[9]。该技术近年来在生物分析领域已得到了广泛应用。
本文建立了PGME生物标志物的纸喷雾质谱测定法，1～2 min内即可完成检测，显著提升时效性，并应用于实际尿液样本分析，为我国确立丙二醇单甲醚生物接触限值提供了数据支撑。
1　材料与方法
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1.1　主要仪器和试剂
LTQ Orbitrap XL线性离子阱静电场轨道阱组合式高分辨质谱仪（美国Thermo Fisher Scientic）；02TMC21三维移动平台（北京集科仪器有限公司）；Xcalibur V2.0数据处理软件（美国ThermoFisher Scientific）；1 CHR色谱纸（英国Whatman）；α-PGME（99.9%，上海阿拉丁生化科技股份有限公司）；2-MPA（99.9%，英国Fluorochem公司)；甲醇（质谱纯，法国Cleman公司）；乙腈、丙酮、乙酸乙酯均为色谱纯（天津市津科生物科技有限责任公司）。
1.2　仪器条件
1.2.1　上样条件
将色谱纸裁剪为高15 mm、底边10 mm的等腰三角形，用平口铜夹固定后，通过三维移动平台调节纸尖端位置，使其正对质谱进样锥孔，且二者间距保持在5 mm左右，见图1。操作时，先取10 μl样品滴加至色谱纸上，静置 1～2 min后，再滴加15 μl甲醇-水（90:10，v/v）喷雾溶剂，同时在纸片后端施加4 kV高压，样品溶解于喷雾溶剂后，在电压作用下于纸尖端形成电喷雾，最终进入质谱进行检测。
1.2.2　质谱条件
采用正离子模式检测α-PGME；喷雾电压为4 kV；离子传输管电压：35 V；离子聚焦透镜电压：110 V；离子传输管温度：275 °C；扫描范围为：20～100 m/z。采用负离子模式检测2-MPA；喷雾电压为-4 kV；离子传输管电压：-35 V；离子聚焦透镜电压：-110 V；离子传输管温度：275 °C；扫描范围为：40～130 m/z。
1.3　样品的采集与保存
以天津市职业病危害因素申报的涂料生产企业为总体，分层随机抽取3家企业，招募50名接触PGME的工人，采集工人尿液用于方法验证。所有受试者均签署知情同意书，本研究获天津市疾病预防控制中心伦理委员会批准（TJCDC-R-2023-016）。参照GBZ/T 295—2017[10]，采集喷漆岗位工人的班末尿50 ml于具盖聚乙烯塑料瓶中，立即测定尿相对密度，弃去相对密度<1.010或>1.030的尿样，冷藏运输。
1.4　混合标准溶液的配制
于10 ml容量瓶中，加入5 ml甲醇，用微量注射器准确加入α-PGME和2-MPA各5.0 μl（20℃时，1 μlα-PGME的质量为0.923 mg，1 μl2-MPA的质量为1.085 mg），用甲醇定容，配制得到浓度为461.5 μg/ml和542.5 μg/ml的α-PGME和2-MPA的混合标准储备液，于4 ℃保存。用甲醇逐级稀释至适宜浓度，得到工作溶液，现用现配；配制尿液样品时，向空白尿液中加入一定量的工作溶液，现用现配。
2　结果
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2.1　质谱条件优化
采用待测物工作溶液在正/负离子模式下进行一级质谱图扫描，同时调整离子传输管温度和电压等质谱参数，使得待测物离子强度最优，从而确定质谱仪条件。依据待测物性质，确定前体离子质荷比（m/z），采集二级质谱图，调节碰撞能量，使待测物产物离子的信号值最高，根据产物离子图，选择相应的定量离子，具体参数见表1。优化后的α-PGME和2-MPA的一级质谱图见图2。
2.2　纸喷雾电离条件的优化
2.2.1　纸基质尖端到质谱进样口的距离优化
以4.62 μg/mlα-PGME和5.42 μg/ml2-MPA标准工作溶液为样品，取10 μl滴加至在纸基质，随后加入10 μl甲醇喷雾溶剂，同时施加4 kV电压，分别在纸尖端到质谱进样口的距离为5、8、10、13和15 mm条件下测定3次，记录响应强度，并绘制距离-响应趋势图，见图3A。结果显示，目标物响应强度随距离增加而降低，5 mm处最高，故选定5 mm为最佳距离。
2.2.2　施加电压优化
以4.62 μg/mlα-PGME和5.42 μg/ml2-MPA标准工作溶液为样品，在纸基质尖端距进样口5 mm条件下，取10 μl进样后滴加10 μl的甲醇溶剂，分别施加2、3、4、5、6和7 kV电压并各测定3次，记录响应强度，绘制电压-响应趋势图，见图3B。结果显示，目标物响应强度随电压升高先增后减，4 kV处达到峰值，故选定4 kV为优化电压。
2.2.3　喷雾溶剂优化
以4.62 μg/mlα-PGME和5.42 μg/ml2-MPA标准工作溶液为样品，在优化的纸尖端距离和电压条件下，分别采用水、甲醇、甲醇/水（90:10，v/v）、乙腈、丙酮、乙酸乙酯作为喷雾溶剂并各测定3次，记录响应强度。尿液基质中2种目标物质响应强度叠加值见图3C。结果显示，甲醇/水（90:10，v/v）和纯甲醇作为喷雾溶剂时效果最佳，信号值强度高，且含水溶剂较纯有机相喷雾持续时间更长。故选择甲醇/水（90:10，v/v）作为喷雾溶剂。
2.2.4　喷雾溶剂用量优化
以4.62 μg/mlα-PGME和5.42 μg/ml2-MPA标准工作溶液为样品，在优化的纸尖端距离、电压及喷雾溶剂条件下，考察5、10、15和20 μl喷雾溶剂用量的响应强度，各测定3次并绘制趋势图，见图3D。结果显示，响应值随喷雾溶剂用量增加先升后趋稳，故选定15 μl为最佳用量。
2.3　标准曲线与检出限、定量限
采用外标法考察α-PGME和2-MPA的线性范围、检出限和定量下限，将不同浓度标准溶液（α-PGME：0.036 92、0.184 6、0.923 0、4.615、23.07和46.15 μg/ml；2-MPA：0.043 40、0.217 0、1.085、5.425、27.12和54.25 μg/ml）分别加入空白尿液后进样，重复测定3次，以响应强度对质量浓度绘制标准曲线。连续测定10个浓度为最低点浓度1/2的加标样品，计算测定浓度的标准差。按3倍标准差计算检出限，10倍标准差算定量下限。α-PGME和2-MPA的线性范围分别为0.036～46.15 μg/ml、0.029～54.25 μg/ml，相关参数见表2。
2.4　方法的精密度与准确度
配制低、中、高浓度加标尿液样品各6份平行样，采用纸喷雾质谱仪测定，计算加标回收率和测定值的精密度。结果显示，α-PGME精密度的相对标准偏差(relative standard deviation，RSD)为3.5%～9.7%，平均加标回收率为93.8%～103.1%；2-MPA的RSD为3.3%～8.0%，平均加标回收率为90.8%～105.0%。见表3。
2.5　样品测定
采用本方法测定天津市50名接触PGME工人的尿液样品，尿相对密度范围均在1.010～1.030范围内。结果显示，α-PGME检出率为18.0%，浓度范围为未检出～1.021 μg/ml；2-MPA的检出率为4.0%，浓度范围为未检出～0.261 μg/ml。
3　讨论
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为评估PGME职业接触人群的内暴露水平，根据其在人体内代谢特性，选择尿液中含量较高的代谢物α-PGME和2-MPA作为研究对象。现有PGME生物标志物的检测方法多为气相色谱法，其依赖繁琐前处理技术，且存在有机溶剂毒性较大、操作步骤冗杂、处理周期长等局限，更无法同时测定两种标志物。纸喷雾质谱法作为新型分析技术，无需或仅需少量前处理即可快速分析生物样本，单个样品平均1～2 min完成检测，较传统方法分析时间缩短95%以上。
在纸喷雾质谱分析中，离子模式的选择显著影响待测物检测与定量，直接关系检测灵敏度、离子化效率及碎裂模式。同时优化离子传输管温度和电压等质谱参数，可提升分析灵敏度、准确性和分辨率，保障目标物高效检测与准确定量。既往研究表明，单一参数调整可能导致目标物回收率与电离抑制发生非一致性变化，需平衡二者关系[11]。本研究以待测物响应值为评估标准，确定各参数最优值。
本研究考察了纸尖端距离、电压、溶剂种类、溶剂用量对α-PGME和2-MPA的响应强度的影响。纸喷雾电离的原理是喷雾溶剂萃取出纸基质上的待测物后，在毛细作用下将含有待测物的溶液运送至纸尖端，在强电场的作用下形成泰勒锥，当其中带电液滴的静电库仑力足以对抗液体的表面张力时，液滴破裂，发生电离，产生离子进入质谱而被检测[12]。纸尖端距离影响离子的传输效率和信号强度。距离过远可能导致离子在空气中扩散和衰减，使灵敏度和分辨率下降，距离过近易导致放电现象，本研究中的α-PGME和2-MPA的最佳距离为5 mm。施加电压的强度会直接影响电离中的传输电流及液体流量，电压过低导致电离不完全，电压过高会产生大量碎片离子，都将使得响应值偏低[13]。本研究中α-PGME和2-MPA的最佳电压为4 kV。喷雾溶剂及其用量对纸喷雾离子源分析性能至关重要，溶剂兼具喷雾与萃取双重作用，不同溶剂对分析物的萃取效果和离子化效率存在差异，因此需选择对分析物溶解性好、表面张力小且易离子化的溶剂[14]。实验结果显示，甲醇/水（90:10，v/v）与纯甲醇作为喷雾溶剂时信号强度最优。溶剂极性和挥发性显著影响分析性能，添加水性溶剂可改善表面张力与喷雾稳定性，延长喷雾时间。因此，本研究选择甲醇/水（90:10，v/v）作为喷雾溶剂以确保尿液样本中信号稳定。鉴于15与20 μl溶剂用量下响应强度无显著差异，为减少消耗，最终选定15 μl。
本研究考察了纸喷雾质谱法的准确度、精密度、检出限、定量限等指标。结果表明，该方法具有准确度高、精密度好、线性范围宽等优势，且无需样品前处理，分析速度快，显著提升了检测时效性，适用于尿液样品的快速筛查。该方法可用于PGME职业接触人群尿液中α-PGME与2-MPA的测定，为我国建立PGME的生物接触限值及检测方法提供依据。
基金
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  • 天津市卫生健康科研项目(TJWJ2023QN087)
  • 天津市医学重点建设学科(TJYXZDXK-3-020B)
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2025年第52卷第19期
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doi: 10.20043/j.cnki.MPM.202505145
  • 接收时间：2025-05-12
  • 首发时间：2026-03-17
  • 出版时间：2025-10-10
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  • 收稿日期：2025-05-12
基金
天津市卫生健康科研项目(TJWJ2023QN087)
天津市医学重点建设学科(TJYXZDXK-3-020B)
作者信息
    1.天津市疾病预防控制中心职业健康所，天津 300011
    2.天津医科大学公共卫生学院

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鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
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红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
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