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Article(id=1216517519184154750, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20250514003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1747152000000, receivedDateStr=2025-05-14, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1767969978376, onlineDateStr=2026-01-09, pubDate=1755187200000, pubDateStr=2025-08-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1767969978376, onlineIssueDateStr=2026-01-09, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1767969978376, creator=13701087609, updateTime=1767969978376, updator=13701087609, issue=Issue{id=1216517514570417012, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='15', 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=null, createTime=1767969977276, creator=13701087609, updateTime=1768211590858, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1217530915467743720, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1217530915467743721, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1216517514570417012, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=264, endPage=272, ext={EN=ArticleExt(id=1216517520085930151, articleId=1216517519184154750, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Determination of 9 kinds of organic acids in fruit and vegetable crisps by liquid chromatography-high resolution mass spectrometry, columnId=1151895321388347923, journalTitle=Journal of Food Safety & Quality, columnName=Food Analysis and Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To establish a method for the determination of 9 kinds of organic acids (oxalic acid, tartaric acid, quinic acid, malic acid, lactic acid, citric acid, fumaric acid, succinic acid and gallic acid) in fruits and vegetable crisps by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Methods The samples were cleaned up by a SAX solid-phase extraction column. The samples were eluted with 0.1% formic acid in water and methanol as mobile phases and separated on an Eclipse Plus C18 column (4.6 mm×250 mm, 5 μm), and monitored in electron spray ionization (ESI)- mode and quantified by external standard method. Results The calibration curves for 9 kinds of organic acids showed good linearity in the concentration range of 50-5000 ng/mL. The correlation coefficients (r) were all greater than 0.9991, the limits of detection (LODs) were in the range of 1-10 ng/mL, the limits of quantitation (LOQs) were in the range of 10-50 ng/mL, the spiked recoveries were in the range of 96%-98%, and the relative standard deviations (RSDs) were in the range of 1.5%-4.9%. Among them, lactic acid [(16.16±0.30) mg/g] and citric acid [(15.53±0.11) mg/g] were the main organic acids in the fruit and vegetable crisps, and their content was significantly higher than those of other organic acids. In contrast, tartaric acid [(1.29±0.11) mg/g] and oxalic acid [(1.56±0.25) mg/g] were found at lower levels. Conclusion The method is simple, rapid, and accurate and can be used for the qualitative and quantitative determination of organic acids (oxalic acid, tartaric acid, quinic acid, malic acid, lactic acid, citric acid, fumaric acid, succinic acid and gallic acid) in fruit and vegetable crumbles as well as other food.

, correspAuthors=Lan-Hua LIU, 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=Lan-Hua LIU, Zhi-Wei SUN, Chang-Qing LIU, Zheng-Qiang WANG, Jing-Cheng ZHANG, Ya-Ling HE, Xue-Feng WU), CN=ArticleExt(id=1216517523135189288, articleId=1216517519184154750, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=液相色谱-高分辨质谱法检测果蔬脆片中9种有机酸含量, 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 建立一种液相色谱-高分辨质谱法(liquid chromatography-high resolution mass spectrometry, LC-HRMS)检测果蔬脆片中9种有机酸(草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸)含量的分析方法。方法 样品经SAX固相萃取柱净化。以0.1%甲酸水和甲醇为流动相等度洗脱, 并经过Eclipse Plus C18色谱柱(4.6 mm×250 mm, 5 μm)分离, 在电喷雾离子源(electron spray ionization, ESI)-模式下进行检测, 并通过外标法进行定量。结果 9种有机酸在质量浓度为50~5000 ng/mL范围内线性关系良好, 相关系数(r)均大于0.9991, 检出限(limits of detection, LODs)在1~10 ng/mL之间, 定量限(limits of quantitation, LOQs)在10~50 ng/mL之间, 加标回收率在96%~98%之间, 相对标准偏差(relative standard deviations, RSDs)在1.5%~4.9%之间。其中, 乳酸[(16.16±0.30) mg/g]和柠檬酸[(15.53±0.11) mg/g]是果蔬脆片中的主要有机酸, 其含量显著高于其他有机酸。相比之下, 酒石酸[(1.29±0.11) mg/g]和草酸[(1.56±0.25) mg/g]含量较低。结论 本研究的方法操作简单, 快速准确, 可用于果蔬脆片以及其他食品中有机酸(草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸)的定性、定量检测。

, correspAuthors=刘兰花, authorNote=null, correspAuthorsNote=
*刘兰花(1983—), 女, 硕士, 工程师, 主要研究方向为食品检测与分析。E-mail:
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Journal of Food Safety & Quality, 2025, 16(12): 224-230., articleTitle=Determination of betamethasone and dexamethasone residues in livestock and poultry products by high performance liquid chromatography-tandem mass spectrometry, refAbstract=null)], funds=[Fund(id=1217127912285848350, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, awardId=2023N06020049, language=CN, fundingSource=安徽省科技项目(2023N06020049), fundOrder=null, country=null), Fund(id=1217127912407483172, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, awardId=202423110050044, language=CN, fundingSource=安徽省科技项目(202423110050044), fundOrder=null, country=null), Fund(id=1217127912541700909, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, awardId=2021054, language=CN, fundingSource=宿州市科技计划项目(2021054), fundOrder=null, country=null), Fund(id=1217127912663335731, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, awardId=SZKJXM202311, language=CN, fundingSource=宿州市科技项目乡村振兴专项(SZKJXM202311), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1217127903532335375, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, xref=1, ext=[AuthorCompanyExt(id=1217127903595249939, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903532335375, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 Instrumental Analysis Center, Hefei University of Technology, Hefei 230009, China), AuthorCompanyExt(id=1217127903607832852, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903532335375, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1 合肥工业大学分析测试中心, 合肥 230009)]), AuthorCompany(id=1217127903733661979, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, xref=2, ext=[AuthorCompanyExt(id=1217127903754633503, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903733661979, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 Anhui Fermented Food Engineering Research Center, College of Food and Bioengineering, Hefei University of Technology, Hefei 230601, China), AuthorCompanyExt(id=1217127903771410721, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903733661979, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2 合肥工业大学食品与生物工程学院, 安徽省发酵食品工程研究中心, 合肥 230601)]), AuthorCompany(id=1217127903888851247, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, xref=3, ext=[AuthorCompanyExt(id=1217127903893045552, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903888851247, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 Anhui Provincial Key Laboratory of Green Manufacturing for Phosgene Industry, Xuancheng 242299, China), AuthorCompanyExt(id=1217127903901434161, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, companyId=1217127903888851247, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 光气产业绿色制造安徽省重点实验室, 宣城 242299)])], figs=[ArticleFig(id=1217127909198840485, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Fig.1, caption=Ion flow chromatograms of 9 kinds of organic acids separated by different mobile phase ratios, figureFileSmall=oJBCWzeZQf4GDJt5UEoCnw==, figureFileBig=gjmAet5+6qstf8boIQ140g==, tableContent=null), ArticleFig(id=1217127909320475309, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=图1, caption=不同流动相比例分离下9种有机酸的离子流色谱图

注: A. 0.1%甲酸水:甲醇=90:10等度洗脱; B. 0.1%甲酸水:甲醇=97:3 (V:V)等度洗脱; C. 0 min, 2%甲醇; 15 min, 20%甲醇; 16 min, 2%甲醇; 20 min, 2%甲醇梯度洗脱; D. 水:甲醇=97:3 (V:V)等度洗脱。

, figureFileSmall=oJBCWzeZQf4GDJt5UEoCnw==, figureFileBig=gjmAet5+6qstf8boIQ140g==, tableContent=null), ArticleFig(id=1217127909475664569, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Fig.2, caption=Ion flow chromatograms of 9 kinds of organic acids separated by different chromatographic columns, figureFileSmall=3lgbH055AffAWczjug8c4Q==, figureFileBig=Y0j24Y3KhkwM49lq4H6mkA==, tableContent=null), ArticleFig(id=1217127909576327873, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=图2, caption=不同色谱柱分离下9种有机酸的离子流色谱图

注: A. ACQUITY UPLC BEH C18色谱柱(500 mm×2.1 mm, 1.7 µm); B. Acclaim RSLC 120 C18色谱柱(100 mm×2.1 mm, 2.2 µm); C. Eclipse Plus C18色谱柱(4.6 mm×250 mm, 5 μm)。

, figureFileSmall=3lgbH055AffAWczjug8c4Q==, figureFileBig=Y0j24Y3KhkwM49lq4H6mkA==, tableContent=null), ArticleFig(id=1217127909668602567, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Fig.3, caption=Ion flow chromatograms of 9 kinds of organic acids separated at different column temperatures, figureFileSmall=ltdF+IqcPUhjqHXfpF8jhQ==, figureFileBig=X+faC7b9+Xn+pxy+Sl+Ktw==, tableContent=null), ArticleFig(id=1217127909815403215, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=图3, caption=不同柱温分离下9种有机酸的离子流色谱图

注: A. 30 ℃; B. 35 ℃; C. 40 ℃; D. 45 ℃。

, figureFileSmall=ltdF+IqcPUhjqHXfpF8jhQ==, figureFileBig=X+faC7b9+Xn+pxy+Sl+Ktw==, tableContent=null), ArticleFig(id=1217127909928649431, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Fig.4, caption=Ion current chromatograms extracted from different organic acids, figureFileSmall=BkPJHIW362SMowRr2SR2mw==, figureFileBig=IKr/H6Fbma3lT48i7z0CNA==, tableContent=null), ArticleFig(id=1217127910016729822, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=图4, caption=不同有机酸提取的离子流色谱图, figureFileSmall=BkPJHIW362SMowRr2SR2mw==, figureFileBig=IKr/H6Fbma3lT48i7z0CNA==, tableContent=null), ArticleFig(id=1217127911337935590, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Table 1, caption=

Parameters of mass spectrometry

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 分子式 分子质量 分子离子[M-H]- 扫描
时间/ms
草酸 C2H2O4 89.9880 88.9880 100
酒石酸 C4H6O6 150.0086 149.0086 100
奎宁酸 C7H12O6 192.0556 191.0556 100
苹果酸 C4H6O5 134.0142 133.0142 100
乳酸 C3H6O3 90.0244 89.0244 100
柠檬酸 C6H8O7 192.0197 191.0197 100
富马酸 C4H4O4 116.0037 115.0037 100
琥珀酸 C4H6O4 118.0193 117.0193 100
没食子酸 C7H6O5 170.0142 169.0142 100
), ArticleFig(id=1217127911472153321, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=表1, caption=

质谱参数

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 分子式 分子质量 分子离子[M-H]- 扫描
时间/ms
草酸 C2H2O4 89.9880 88.9880 100
酒石酸 C4H6O6 150.0086 149.0086 100
奎宁酸 C7H12O6 192.0556 191.0556 100
苹果酸 C4H6O5 134.0142 133.0142 100
乳酸 C3H6O3 90.0244 89.0244 100
柠檬酸 C6H8O7 192.0197 191.0197 100
富马酸 C4H4O4 116.0037 115.0037 100
琥珀酸 C4H6O4 118.0193 117.0193 100
没食子酸 C7H6O5 170.0142 169.0142 100
), ArticleFig(id=1217127911618953972, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Table 2, caption=

Linear equations, limits of detection and limits of quantification of organic acids

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 线性范围/(ng/mL) 标准曲线方程 r 检出限/(ng/mL) 定量限/(ng/mL)
草酸 50~5000 Y=1.29e7X+1.97e5 0.9993 10 50
酒石酸 50~5000 Y=2.99e8X+5.88e6 0.9994 1 10
奎宁酸 50~5000 Y=2.68e8X+3.04e7 0.9993 1 10
苹果酸 50~5000 Y=1.59e8X+8.35e6 0.9995 1 10
乳酸 50~5000 Y=2.26e8X+3.60e7 0.9994 1 10
柠檬酸 50~5000 Y=2.36e8X-6.24e6 1.0000 1 10
富马酸 50~5000 Y=5.31e7X+2.99e5 1.0000 10 20
琥珀酸 50~5000 Y=1.64e8X+1.09e7 0.9991 1 10
没食子酸 50~5000 Y=4.47e8X+2.99e7 0.9994 1 10
), ArticleFig(id=1217127911744783101, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=表2, caption=

有机酸线性方程、检出限和定量限

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 线性范围/(ng/mL) 标准曲线方程 r 检出限/(ng/mL) 定量限/(ng/mL)
草酸 50~5000 Y=1.29e7X+1.97e5 0.9993 10 50
酒石酸 50~5000 Y=2.99e8X+5.88e6 0.9994 1 10
奎宁酸 50~5000 Y=2.68e8X+3.04e7 0.9993 1 10
苹果酸 50~5000 Y=1.59e8X+8.35e6 0.9995 1 10
乳酸 50~5000 Y=2.26e8X+3.60e7 0.9994 1 10
柠檬酸 50~5000 Y=2.36e8X-6.24e6 1.0000 1 10
富马酸 50~5000 Y=5.31e7X+2.99e5 1.0000 10 20
琥珀酸 50~5000 Y=1.64e8X+1.09e7 0.9991 1 10
没食子酸 50~5000 Y=4.47e8X+2.99e7 0.9994 1 10
), ArticleFig(id=1217127911870612227, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=EN, label=Table 3, caption=

Organic acid content, spiked recovery rates and RSDs of fruit and vegetable crisps

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 保留时间/min 含量
/(mg/g)		 加标
回收率/%		 RSDs
/%
草酸 2.70 1.56±0.25 97 2.5
酒石酸 2.92 1.29±0.11 97 3.3
奎宁酸 2.96 4.27±0.06 96 1.5
苹果酸 3.42 11.76±0.33 96 1.6
乳酸 4.01 16.16±0.30 97 4.9
柠檬酸 5.11 15.53±0.11 98 2.7
富马酸 5.63 1.88±0.26 98 4.3
琥珀酸 6.12 2.68±0.32 97 4.4
没食子酸 9.58 1.68±0.32 96 3.1
), ArticleFig(id=1217127912017412877, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1216517519184154750, language=CN, label=表3, caption=

果蔬脆片有机酸含量、加标回收率与RSDs

, figureFileSmall=null, figureFileBig=null, tableContent=
成分 保留时间/min 含量
/(mg/g)		 加标
回收率/%		 RSDs
/%
草酸 2.70 1.56±0.25 97 2.5
酒石酸 2.92 1.29±0.11 97 3.3
奎宁酸 2.96 4.27±0.06 96 1.5
苹果酸 3.42 11.76±0.33 96 1.6
乳酸 4.01 16.16±0.30 97 4.9
柠檬酸 5.11 15.53±0.11 98 2.7
富马酸 5.63 1.88±0.26 98 4.3
琥珀酸 6.12 2.68±0.32 97 4.4
没食子酸 9.58 1.68±0.32 96 3.1
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液相色谱-高分辨质谱法检测果蔬脆片中9种有机酸含量
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刘兰花 1, * , 孙智伟 2 , 刘长庆 3 , 王争强 3 , 张竞成 1 , 何雅玲 1 , 吴学凤 2
食品安全质量检测学报 | 食品分析与检测 2025,16(15): 264-272
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食品安全质量检测学报 | 食品分析与检测 2025, 16(15): 264-272
液相色谱-高分辨质谱法检测果蔬脆片中9种有机酸含量
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刘兰花1, * , 孙智伟2, 刘长庆3, 王争强3, 张竞成1, 何雅玲1, 吴学凤2
作者信息
  • 1 合肥工业大学分析测试中心, 合肥 230009
  • 2 合肥工业大学食品与生物工程学院, 安徽省发酵食品工程研究中心, 合肥 230601
  • 3 光气产业绿色制造安徽省重点实验室, 宣城 242299

通讯作者:

*刘兰花(1983—), 女, 硕士, 工程师, 主要研究方向为食品检测与分析。E-mail:
Determination of 9 kinds of organic acids in fruit and vegetable crisps by liquid chromatography-high resolution mass spectrometry
Lan-Hua LIU1, * , Zhi-Wei SUN2, Chang-Qing LIU3, Zheng-Qiang WANG3, Jing-Cheng ZHANG1, Ya-Ling HE1, Xue-Feng WU2
Affiliations
  • 1 Instrumental Analysis Center, Hefei University of Technology, Hefei 230009, China
  • 2 Anhui Fermented Food Engineering Research Center, College of Food and Bioengineering, Hefei University of Technology, Hefei 230601, China
  • 3 Anhui Provincial Key Laboratory of Green Manufacturing for Phosgene Industry, Xuancheng 242299, China
出版时间: 2025-08-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20250514003
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摘要
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目的 建立一种液相色谱-高分辨质谱法(liquid chromatography-high resolution mass spectrometry, LC-HRMS)检测果蔬脆片中9种有机酸(草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸)含量的分析方法。方法 样品经SAX固相萃取柱净化。以0.1%甲酸水和甲醇为流动相等度洗脱, 并经过Eclipse Plus C18色谱柱(4.6 mm×250 mm, 5 μm)分离, 在电喷雾离子源(electron spray ionization, ESI)-模式下进行检测, 并通过外标法进行定量。结果 9种有机酸在质量浓度为50~5000 ng/mL范围内线性关系良好, 相关系数(r)均大于0.9991, 检出限(limits of detection, LODs)在1~10 ng/mL之间, 定量限(limits of quantitation, LOQs)在10~50 ng/mL之间, 加标回收率在96%~98%之间, 相对标准偏差(relative standard deviations, RSDs)在1.5%~4.9%之间。其中, 乳酸[(16.16±0.30) mg/g]和柠檬酸[(15.53±0.11) mg/g]是果蔬脆片中的主要有机酸, 其含量显著高于其他有机酸。相比之下, 酒石酸[(1.29±0.11) mg/g]和草酸[(1.56±0.25) mg/g]含量较低。结论 本研究的方法操作简单, 快速准确, 可用于果蔬脆片以及其他食品中有机酸(草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸)的定性、定量检测。

液相色谱-高分辨质谱法  /  果蔬脆片  /  有机酸  /  固相萃取柱

Objective To establish a method for the determination of 9 kinds of organic acids (oxalic acid, tartaric acid, quinic acid, malic acid, lactic acid, citric acid, fumaric acid, succinic acid and gallic acid) in fruits and vegetable crisps by liquid chromatography-high resolution mass spectrometry (LC-HRMS). Methods The samples were cleaned up by a SAX solid-phase extraction column. The samples were eluted with 0.1% formic acid in water and methanol as mobile phases and separated on an Eclipse Plus C18 column (4.6 mm×250 mm, 5 μm), and monitored in electron spray ionization (ESI)- mode and quantified by external standard method. Results The calibration curves for 9 kinds of organic acids showed good linearity in the concentration range of 50-5000 ng/mL. The correlation coefficients (r) were all greater than 0.9991, the limits of detection (LODs) were in the range of 1-10 ng/mL, the limits of quantitation (LOQs) were in the range of 10-50 ng/mL, the spiked recoveries were in the range of 96%-98%, and the relative standard deviations (RSDs) were in the range of 1.5%-4.9%. Among them, lactic acid [(16.16±0.30) mg/g] and citric acid [(15.53±0.11) mg/g] were the main organic acids in the fruit and vegetable crisps, and their content was significantly higher than those of other organic acids. In contrast, tartaric acid [(1.29±0.11) mg/g] and oxalic acid [(1.56±0.25) mg/g] were found at lower levels. Conclusion The method is simple, rapid, and accurate and can be used for the qualitative and quantitative determination of organic acids (oxalic acid, tartaric acid, quinic acid, malic acid, lactic acid, citric acid, fumaric acid, succinic acid and gallic acid) in fruit and vegetable crumbles as well as other food.

liquid chromatography-high resolution mass spectrometry  /  fruit and vegetable crisps  /  organic acids  /  solid-phase extraction columns
刘兰花, 孙智伟, 刘长庆, 王争强, 张竞成, 何雅玲, 吴学凤. 液相色谱-高分辨质谱法检测果蔬脆片中9种有机酸含量. 食品安全质量检测学报, 2025 , 16 (15) : 264 -272 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250514003
Lan-Hua LIU, Zhi-Wei SUN, Chang-Qing LIU, Zheng-Qiang WANG, Jing-Cheng ZHANG, Ya-Ling HE, Xue-Feng WU. Determination of 9 kinds of organic acids in fruit and vegetable crisps by liquid chromatography-high resolution mass spectrometry[J]. Journal of Food Safety & Quality, 2025 , 16 (15) : 264 -272 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20250514003
正文
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0 引言
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有机酸是食品中重要的天然成分之一, 对食品的风味、保鲜和营养价值具有重要作用[1-3]。常见的食品有机酸包括柠檬酸、苹果酸、乳酸、醋酸和酒石酸等, 它们广泛存在于水果、蔬菜、乳制品及发酵食品中[4-5]。有机酸赋予产品独特的酸味, 同时平衡甜味, 增强食品的口感。有机酸还具有显著的防腐作用, 通过降低食品的pH抑制微生物生长, 从而延长食品的保质期[6-7]。此外, 有机酸还具有一定的营养价值, 参与人体代谢, 如柠檬酸循环, 为人体提供能量[8-9]。同时, 它们可促进矿物质吸收, 如乳酸可提高钙的生物利用率。在食品加工中, 有机酸常作为酸度调节剂、抗氧化剂、增香剂和防腐剂, 是现代食品工业不可或缺的功能成分[10]。
有机酸的分析方法主要包括气相色谱法和气相色谱-质谱法、离子色谱法、高效液相色谱法和高效液相色谱-质谱法[2-3]。气相色谱主要用于测定低分子质量, 易挥发的有机酸。对于沸点较高的有机酸通常需要衍生化处理, 但衍生化过程会增加操作难度和前处理时间, 并且可能导致误差加剧, 从而限制了气相色谱法和气相色谱-质谱法的应用[11]。离子色谱法和高效液相色谱法是有机酸分析最常用的方法, 但非特异性检测器易受到样品基质的干扰[12]。因此对于基质复杂的样品, 往往需要采用相对复杂的前处理去除干扰, 以保证分析的灵敏度和准确度[5,13]。
质谱法具有高选择性、高灵敏度等优点, 可弥补液相色谱法定性能力不足的缺点[14-15]。高效液相色谱-质谱法也因此成为目前公认的有机酸高效检测方法, 而高分辨质谱以其精准识别、精确定量的技术优势, 在食品分析领域受到了越来越多的关注[16-17]。本研究通过建立液相色谱-高分辨质谱法对食品中9种有机酸(草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸、没食子酸)直接进行定量检测, 以期为果蔬脆片及食品中有机酸的检测提供新的思路。
1 材料与方法
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1.1 材料与试剂
罐装12种水果蔬菜综合果蔬脆片(商品编号10141170389640, 安徽东方果园生物科技有限公司)。
草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸标准品(纯度99%, 上海源叶生物科技有限公司); 甲醇、甲酸、乙腈、乙醇(分析纯, 国药集团化学试剂有限公司)。
1.2 仪器与设备
LT2002E电子天平(精度0.00001 g, 常熟天量仪器有限责任公司); Millipore A10超纯水机(美国密理博公司); DMV-16涡旋混匀仪(广东科寅设备有限公司); RE100-pro型旋转蒸发仪(北京大龙兴创实验仪器股份公司); Vanquish Q Exactive Plus液相色谱-四极杆静电场轨道阱质谱仪(美国赛默飞世尔科技有限公司); Eclipse Plus C18色谱柱(4.6 mm× 250 mm, 5 μm)(美国安捷伦有限公司); SAX固相萃取柱(武汉艾逗生物科技有限公司)。
1.3 实验方法
1.3.1 标准溶液的配制
混合标准储备溶液分别准确称取10.00 mg的草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸、没食子酸的标准品于9个烧杯中, 用10%甲醇水溶液溶解并定容至10 mL容量瓶中, 摇匀, 得到1 mg/mL的各单标储备液。取各单标储备液各1 mL于10 mL容量瓶中, 用10%甲醇水溶液溶解并定容至刻度, 摇匀, 得到0.1 mg/mL的混合标准储备溶液。临用前将混合有机酸储备液用10%的甲醇水溶液依次稀释至0.050、0.010、0.005、0.001 mg/mL。
1.3.2 样品的前处理
将果蔬脆片粉碎、搅拌均匀后, 称取5 g(精确至0.01 g)于50 mL的离心管中, 加入20 mL超纯水后, 将离心管置于均质机中, 以2000 r/min的速度均质提取5 min, 并于高速离心机在10000 r/min的转速下离心5 min, 分离得上清液和残渣。将残渣重复以上的操作, 并收集所有上清液混合。随后在上清液中加入等体积无水乙醇并定容至50 mL, 摇匀, 得上清溶液。将10 mL上清溶液加入100 mL鸡心瓶中, 再加入10 mL无水乙醇, 在80 ℃±2 ℃下旋转浓缩至近干时, 再加入5 mL无水乙醇继续浓缩至彻底干燥, 依次用1 mL超纯水洗涤鸡心瓶两次后, 转移至经过预活化(1 mL甲醇淋洗3次, 1 mL超纯水淋洗3次)的SAX固相萃取柱中, 流速为1 mL/min, 弃去流出液, 用5 mL超纯水淋洗净化柱, 再用5 mL甲酸甲醇溶液洗脱, 流速为1 mL/min, 收集洗脱液于50 mL离心管中, 氮吹至近干后, 用5 mL体积分数为0.1%的甲酸水溶液振荡溶解残渣, 用0.22 μm的水相滤膜过滤, 得果蔬脆片待测液。
1.4 仪器条件
1.4.1 色谱条件
色谱柱: Eclipse Plus C18 (4.6 mm×250 mm, 5 μm)。流动相: 甲醇和体积分数为0.1%的甲酸水溶液按照体积比3:97混合。洗脱流速: 0.8 mL/min; 柱温: 30 ℃; 进样体积: 10 μL。
1.4.2 质谱条件
电喷雾离子源(electron spray ionization, ESI); 喷雾电压: 3200 V; 蒸发温度: 400 ℃; 离子传输温度: 500 ℃; 鞘气: 50 arb; 辅助气: 20 arb; 扫描方式: 负离子; 扫描范围: 50~750 m/z。
1.5 数据处理
LC-HRMS配有Xcalibur数据处理软件, 通过该软件建立标准曲线, 计算结果, 采用Microsoft Excel 2009处理数据及绘制图表。谱图由Xcalibur导出原图, 并通过Photoshop作图软件组图绘制。本研究中净化柱的选择和回收率均采用6个平行样品计算。
食品待测液有机酸含量按公式(1)计算:
X=$\frac{C\times V}{m}$
式中: X为食品待测液中有机酸的含量, mg/g; C为由标准曲线求得食品待测液中某有机酸的质量浓度, mg/mL; V为食品待测液定容体积, mL; m为最终食品待测液代表的食品质量, g。
有机酸加标回收率按公式(2)计算:
P/%=$\frac{{C}_{2}-{C}_{1}}{{C}_{3}}$×100%
式中: P为回收率, %; C2: 加标食品待测液测定值, mg/mL; C1: 食品待测液测定值, mg/mL; C3: 加标量, mg/mL。
2 结果与分析
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2.1 液相条件的确定
通过调整流动相的比例, 使各有机酸分离。如图1A、B所示, 这9种有机酸极性较强, 在色谱柱上的保留较小, 随着水相溶剂(0.1%甲酸)含量的增加, 分离效果变好[18-19]。同时比较了纯水溶剂和0.1%甲酸水在相同比例下的分离情况, 由图1B、D可见, 添加了甲酸的水相有机相分离度更好些, 在ESI-模式下, 流动相中加入适量甲酸可增强待测组分离子化效率, 提高检测响应强度[20]。甲醇比乙腈在有机酸方面有更强的洗脱能力, 可将保留时间较早的色谱峰有效分离, 因此选择甲醇和0.1%甲酸水作为流动相[21]。采用梯度洗脱和等度洗脱方式进行比较, 如图1C所示, 发现梯度洗脱也可达到分离9种有机酸的实验目的, 但是梯度洗脱基线波动加大, 不利于低浓度样品的信号的采集。故0.1%甲酸水:甲醇=97:3 (V:V)等度洗脱为最佳色谱分离条件。
2.2 质谱条件的确定
将质量浓度为1 mg/mL的标准液样品引入静电场离子轨道阱质谱仪(Orbitrap MS)。在分析过程中, Orbitrap MS通过施加静电场捕获负离子, 并使其在轨道井中做谐振运动, 从而测量离子的振动频率。由于离子的振动频率与其质荷比(m/z)之间存在直接关系, 因此该方法能够实现有机酸负离子的高精度检测和分离[22-23]。在负离子模式下, 羧基等酸性基团的去质子化效应得到增强, 从而提高有机酸在质谱中的响应信号, 便于精确分析其分子量和结构信息[24]。通过选择ESI负离子模式进行质谱扫描, 最终获得目标物的离子对信息及相关质谱参数, 如表1所示。
2.3 色谱柱的确定
有机酸在不同色谱柱下的分离效果如图2所示。图2A和B中的多个色谱峰重叠较为严重, 尤其是在色谱图的前段(保留时间<5 min)。与之相比, 图2C中的峰分离度明显提高, 尤其在前段和中段, 表现出更好的分离性能。此外, 图2C中的色谱峰显示出较好的对称性, 而图2A和2B中的部分峰则出现了较为明显的宽峰或拖尾现象。图2C所使用的色谱柱(Eclipse Plus C18 column)虽然粒径较大, 但其较长的柱长提供了较高的理论塔板数, 从而提高了分离效率。相比之下, 图2A和B所用的色谱柱粒径较小(分别为1.7 μm和2.2 μm), 通常需要更高的柱压。图2C的色谱柱粒径为5 μm, 在常规高效液相系统中更易使用且具有更好的稳定性。此外, 图2C的色谱柱内径较宽(4.6 mm), 更适合负载较大样品量的分离分析。
虽然3种色谱柱的总分析时间均控制在15 min以内, 但图2C在不显著延长分析时间的情况下, 提供了更优的分离效果和峰形。因此, 选择Eclipse Plus C18色谱柱作为有机酸分离的柱子。
2.4 柱温的确定
通过对比不同柱温下的色谱图, 可以发现较低的柱温(如图3A)下, 各个峰之间的间隔明显较大, 尤其是早期保留时间(<10 min)的低分子有机酸, 其与相邻峰的分离度最佳。图3A中, 色谱峰形相对对称且尖锐, 特别是关键的前段峰(如2.89 min和3.10 min)。然而, 较高柱温(如图3B和3C)会导致峰的宽度缩小, 且部分峰出现重叠。随着柱温的升高, 峰形略有展宽或出现拖尾现象, 这表明低温下柱效更高。在30 ℃时, 各峰的保留时间稳定且间隔适中, 这种保留时间分布有利于定量分析, 增强了方法的可重复性。相比之下, 在更高温度(如图3C和D)下, 部分保留时间较短的峰出现了轻微的压缩。柱温较低时(30 ℃), 分离选择性更好, 各有机酸分子之间的极性差异得以更有效地体现, 导致峰的分离更为清晰和明确。随着温度升高(35、40、45 ℃), 峰的选择性稍有减弱, 部分有机酸可能对高温较为敏感, 在高温下发生部分降解或分解反应。综合来看, 30 ℃的低温操作有助于保持样品的化学稳定性[25]。因此, 最终选择30 ℃作为有机酸分离时的柱温。
2.5 标准曲线、检出限与定量限
图4展示了不同有机酸提取的离子流色谱图及其保留时间。由表2可知, 9种有机酸在50~5000 ng/mL质量浓度范围内, 相关系数(r)>0.9991, 计算酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、琥珀酸、没食子酸的检出限均为1 ng/mL, 定量限为10 ng/mL。草酸、富马酸的检出限均为10 ng/mL, 定量限分别为50 ng/mL和20 ng/mL, 与部分已有方法相比检出限和定量限更低, 定量更为精准可靠[26-27]。
2.6 回收率与精密度以及实际样品测定
表3展示了果蔬脆片中9种有机酸的分子离子质量、保留时间、含量、加标回收率及RSDs。乳酸[(16.16±0.30) mg/g]和柠檬酸[(15.53±0.11) mg/g]是果蔬脆片中的主要有机酸, 其含量高于其他化合物, 表明这两种酸是样品中的主要成分。相比之下, 酒石酸[(1.29±0.11) mg/g]和草酸[(1.56±0.25) mg/g]含量较低, 这可能是由于其在果蔬原料中的天然含量较少[28-29]。加标回收率在96%~98%之间, 均处于95%~105%的合理范围内, 表明该检测方法具有较高的准确性。柠檬酸和富马酸的回收率最高(98%), 显示其检测精度优良。RSDs均小于5%, 在1.5%(奎宁酸)到4.9%(乳酸)之间, 表明该方法具有良好的精密性。乳酸的较高RSDs可能是由于其含量较高, 且在提取过程中存在微小波动所致[30-31]。
综上所述, 表3中的数据表明该分析方法能够快速、准确地分离和定量果蔬脆片中的9种有机酸。该方法的高分离度、准确性和精密性为果蔬功能成分的定量研究提供了有力支持, 也为质量控制提供了可靠依据。
3 讨论和结论
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该方法有效去除了基质干扰, 但在复杂食品基质中, 仍可能存在一些未知的干扰物质, 可能影响分析结果的准确性。未来可以通过进一步优化固相萃取柱的选择和样品前处理步骤, 或者尝试其他净化技术(如离子交换、溶剂萃取等), 以进一步提高方法的抗干扰能力[32-33]。虽然该方法已在果蔬脆片中得到了验证, 但其在其他类型食品中的应用范围仍需进一步拓展。不同食品的成分差异可能对分析结果产生影响, 因此, 需要对该方法在不同食品中的适用性和稳定性进行更多的验证, 以确保其具有广泛的适用性。随着技术的发展, 更多创新性的样品前处理和检测方法将不断涌现, 为食品行业提供更强有力的技术支持。通过不断优化现有方法和拓宽应用领域, 能够有效提升食品安全监管水平, 保障消费者的健康。
本研究建立了SAX固相萃取柱净化, 液相色谱-高分辨质谱法检测果蔬脆片中草酸、酒石酸、奎宁酸、苹果酸、乳酸、柠檬酸、富马酸、琥珀酸和没食子酸9种有机酸的方法, 并对仪器条件包括液相条件、色谱柱选择、柱温选择条件进行了优化。该方法能有效地除去基质干扰, 检出限为1~10 ng/mL, 定量限为10~50 ng/mL, 加标回收率在96%~98%之间, RSDs为1.5%~4.9%。该方法方便快速、准确可靠, 满足实际样品检测要求, 可为果蔬脆片等其他食品中有机酸的监测提供技术支撑, 有助于对食品行业中各类食品有机酸的快速定量和解析。
基金
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  • 安徽省科技项目(2023N06020049)
  • 安徽省科技项目(202423110050044)
  • 宿州市科技计划项目(2021054)
  • 宿州市科技项目乡村振兴专项(SZKJXM202311)
文献
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2025年第16卷第15期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20250514003
  • 接收时间：2025-05-14
  • 首发时间：2026-01-09
  • 出版时间：2025-08-15
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  • 收稿日期：2025-05-14
基金
安徽省科技项目(2023N06020049)
安徽省科技项目(202423110050044)
宿州市科技计划项目(2021054)
宿州市科技项目乡村振兴专项(SZKJXM202311)
作者信息
    1 合肥工业大学分析测试中心, 合肥 230009
    2 合肥工业大学食品与生物工程学院, 安徽省发酵食品工程研究中心, 合肥 230601
    3 光气产业绿色制造安徽省重点实验室, 宣城 242299

通讯作者:

*刘兰花(1983—), 女, 硕士, 工程师, 主要研究方向为食品检测与分析。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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