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Article(id=1153986714193355239, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20241021003, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1729440000000, receivedDateStr=2024-10-21, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1753061472700, onlineDateStr=2025-07-21, pubDate=1737734400000, pubDateStr=2025-01-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1753061472700, onlineIssueDateStr=2025-07-21, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1753061472700, creator=13701087609, updateTime=1753061472700, updator=13701087609, issue=Issue{id=1153986709126635984, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='2', pageStart='1', pageEnd='324', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1753061471492, creator=13701087609, updateTime=1760345674980, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1184538872999457117, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1184538872999457118, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153986709126635984, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=127, endPage=137, ext={EN=ArticleExt(id=1153986715376148976, articleId=1153986714193355239, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Effects of different processing methods on flavor substances of beef jerky products, columnId=1153986712016506967, journalTitle=Journal of Food Safety & Quality, columnName=Special Topic: Research on Optimization and Application of Food Processing Technology, runingTitle=null, highlight=null, articleAbstract=

Objective To study the characteristics of volatile flavor substances in beef jerky products processed by different processing methods. Methods Beef was processed by different processing methods (15 kinds of curing, bacterial curing, air-drying, etc.), and analyzed by headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography-time of flight mass spectrometry. Results The differential metabolites in 15 kinds of samples were detected. Among them, during the pickling process of Staphylococcus xylosus, Lactobacillus plantarum and Pediococcus pentosaceus, the significant differential metabolites included 12 kinds of sugars, acids and esters. The cured beef processed by air-drying method and the cured beef with bacteria produced 8 kinds of extremely significant different metabolites such as maltose, decanoic acid and orotic acid after air-drying. More than 11 kinds of differential metabolites including naphthalene, 3-hydroxy-DL-norvaline, L-menthone, n-decanoic acid, D-pinobiose and myristic acid were detected in the cured beef processed by normal baking than in the cured beef processed by vacuum baking. The metabolic pathways involved in vacuum baking were citric acid cycle, alanine, aspartic acid and glutamic acid metabolism, and the metabolic pathways involved in baking were amino sugar and nucleotide sugar metabolism. Conclusion In this study, the changes of volatile flavor substances in beef jerky products with different processing methods are analyzed, which provide methodological guidance for improving the quality and taste of air-drying beef.

, correspAuthors=Fan YANG, 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=Mei-Ren WANG, Bo-Hui WANG, Fan YANG, Xue-Qin LIU, Chun-Xia SU, Xiao-Rong DUAN), CN=ArticleExt(id=1153986764256567505, articleId=1153986714193355239, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=不同加工方式对牛肉干产品风味物质的影响, columnId=1153986712150724698, journalTitle=食品安全质量检测学报, columnName=本期专题：食品加工工艺优化及应用研究, runingTitle=null, highlight=null, articleAbstract=

目的 研究不同加工处理方式牛肉干产品挥发性风味物质的特征。方法 将牛肉用不同方式(腌制、加菌腌制、风干等15种)进行加工, 对其进行顶空固相微萃取结合全二维气相色谱-飞行时间质谱分析。结果 检测15种样品中的差异代谢物质, 其中添加木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制的过程中, 显著性差异代谢物质包含糖、酸、酯类等共12种; 以风干方式加工的腌制牛肉和加菌腌制牛肉, 在风干后产生麦芽糖、癸酸和乳清酸等8种极显著差异代谢物; 以正常烘烤方式加工的腌制牛肉比真空烘烤加工的腌制牛肉多检出包含萘、3-羟基-DL-正缬氨酸、左旋薄荷酮、正癸酸、D-松二糖、肉豆蔻酸在内的11种差异代谢物, 并且真空烘烤参与的代谢通路有柠檬酸循环、丙氨酸、天冬氨酸和谷氨酸代谢等, 烘烤参与的代谢通路为氨基糖和核苷酸糖代谢。结论 本研究分析了不同加工处理方式的牛肉干产品挥发性风味物质的变化规律, 为提升风干牛肉的品质和口感提供了方法指导。

, correspAuthors=杨帆, authorNote=null, correspAuthorsNote=
* 杨帆(1971—), 男, 硕士, 教授, 主要研究方向为食品加工与安全方向。E-mail:
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王美仁(1985—), 女, 硕士, 讲师, 实验师, 主要研究方向为食品加工与安全。E-mail:

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王美仁(1985—), 女, 硕士, 讲师, 实验师, 主要研究方向为食品加工与安全。E-mail:

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注: std.dev表示标准偏差。

, figureFileSmall=HORqNCor0C4GFAAnosVAYA==, figureFileBig=DXsU6zrcJV23DNe+R9Ad9Q==, tableContent=null), ArticleFig(id=1184567072332854028, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714193355239, language=EN, label=Fig.4, caption=Principal component analysis scatter plot, figureFileSmall=uTtXi2uVCWQ+0D8A64BXMQ==, figureFileBig=WJBLCgV/EiTvOR3pvQJMSg==, tableContent=null), ArticleFig(id=1184567072391574286, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714193355239, language=CN, label=图4, caption=主成分分析散点图

注: a. 主成分分析模型三维得分散点图; b. 全部样本主成分分析得分散点图。

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注: 图中横坐标代表不同样本分组, 纵坐标代表所有的代谢物, 不同位置的色块代表对应位置代谢物的相对表达量, 其中红色表示该物质含量高表达, 蓝色表示该物质含量低表达。

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Internal standard peak area

, figureFileSmall=null, figureFileBig=null, tableContent=
样品 峰面积 RSDs/%
QC1 9953895.4 5.56
QC2 10871511.9
QC3 10624968.8
QC4 11075960.5
QC5 11531257.6
QC6 10112275.8
平均值 10694678.3
), ArticleFig(id=1184567073339487016, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714193355239, language=CN, label=表1, caption=

内标峰面积

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样品 峰面积 RSDs/%
QC1 9953895.4 5.56
QC2 10871511.9
QC3 10624968.8
QC4 11075960.5
QC5 11531257.6
QC6 10112275.8
平均值 10694678.3
), ArticleFig(id=1184567073406595882, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714193355239, language=EN, label=Table 2, caption=

Screening of differential metabolites in different groups

, figureFileSmall=null, figureFileBig=null, tableContent=
组别 中文名称 英文名称 VIP P
C vs B 1,2-环己二酮 1,2-cyclohexanedione 4.10 0.00
癸酸 capric acid 3.97 0.00
核糖酸, γ-内酯 ribonic acid, gamma-lactone 1.02 0.00
乳清酸 orotic acid 3.93 0.00
2-脱氧-D-葡萄糖 2-deoxy-D-glucose 3.73 0.01
葡萄糖内酯 gluconic lactone 1.13 0.00
环氧缩酮 conduritol b epoxide 2.27 0.00
葡萄糖酸 gluconic acid 1.15 0.00
龙胆糖 gentiobiose 3.84 0.00
半乳糖 melibiose 1.03 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.35 0.00
麦芽糖 maltotriose 4.16 0.00
E vs D 5-氨基戊酸内酰胺 5-aminovaleric acid lactam 3.61 0.00
癸酸 capric acid 1.62 0.01
乳清酸 orotic acid 3.38 0.00
龙胆糖 gentiobiose 4.11 0.00
半乳糖 melibiose 1.07 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.02 0.00
酵母醇 zymosterol 3.14 0.01
麦芽糖 maltotriose 3.61 0.00
K vs J 苄氧羰基-L-亮氨酸脱氨酶 carbobenzyloxy-L-leucine degr 3.85 0.01
2-甲基戊二酸 2-methylglutaric acid 2.70 0.03
甲酮戊二酸 alpha-ketoglutaric acid 1.77 0.01
3-苯基乳酸 3-phenyllactic acid 1.75 0.05
半乳糖 xylose 2.31 0.01
乳清酸 orotic acid 3.72 0.00
肉豆蔻酸 myristic acid 1.03 0.00
6-磷酸葡萄糖 glucose-6-phosphate 3.63 0.03
龙胆糖 gentiobiose 4.24 0.00
I vs H 萘 naphthalene 3.47 0.00
3-羟基缬氨酸 3-hydroxynorvaline 1.39 0.05
癸酸 capric acid 1.52 0.01
薄荷酮 menthone 1.14 0.00
乳清酸 orotic acid 3.11 0.00
金合欢醛 farnesal 1.61 0.00
葡萄糖 glucose 1.40 0.00
6-磷酸果糖 fructose-6-phosphate 1.56 0.00
松二糖 turanose 2.98 0.00
龙胆糖 gentiobiose 2.79 0.00
O vs N 甘氨酸 glycine 1.15 0.00
草酸 oxalic acid 2.67 0.00
萘 naphthalene 2.99 0.00
2-丁炔-1,4-二醇 2-butyne-1,4-diol 1.98 0.00
环己烷-1,2-二醇 cyclohexane-1,2-diol 3.14 0.00
二氢香芹醇 (-)-dihydrocarveol 2.19 0.00
1,4-环己二酮 1,4-cyclohexanedione 1.92 0.05
1-茚醇 1-indanol 2.93 0.00
2-甲基戊二酸 2-methylglutaric acid 2.79 0.00
癸酸 capric acid 2.08 0.01
N-乙基马来酰胺酸 N-ethylmaleamic acid 1.86 0.01
天冬氨酸 aspartic acid 1.49 0.03
薄荷酮 menthone 3.16 0.00
阿洛糖 allose 1.30 0.00
乳清酸 orotic acid 2.67 0.00
反式-4-羟基-L-脯氨酸 trans-4-hydroxy-L-proline 1.17 0.03
金合欢醛 farnesal 2.72 0.00
葡萄糖 glucose 2.53 0.00
异肌醇 allo-inositol 1.25 0.03
槐糖 sophorose 2.18 0.03
龙胆糖 gentiobiose 2.39 0.00
半乳糖 melibiose 1.16 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.58 0.00
), ArticleFig(id=1184567073494676268, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153986714193355239, language=CN, label=表2, caption=

不同组别差异代谢物筛选

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组别 中文名称 英文名称 VIP P
C vs B 1,2-环己二酮 1,2-cyclohexanedione 4.10 0.00
癸酸 capric acid 3.97 0.00
核糖酸, γ-内酯 ribonic acid, gamma-lactone 1.02 0.00
乳清酸 orotic acid 3.93 0.00
2-脱氧-D-葡萄糖 2-deoxy-D-glucose 3.73 0.01
葡萄糖内酯 gluconic lactone 1.13 0.00
环氧缩酮 conduritol b epoxide 2.27 0.00
葡萄糖酸 gluconic acid 1.15 0.00
龙胆糖 gentiobiose 3.84 0.00
半乳糖 melibiose 1.03 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.35 0.00
麦芽糖 maltotriose 4.16 0.00
E vs D 5-氨基戊酸内酰胺 5-aminovaleric acid lactam 3.61 0.00
癸酸 capric acid 1.62 0.01
乳清酸 orotic acid 3.38 0.00
龙胆糖 gentiobiose 4.11 0.00
半乳糖 melibiose 1.07 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.02 0.00
酵母醇 zymosterol 3.14 0.01
麦芽糖 maltotriose 3.61 0.00
K vs J 苄氧羰基-L-亮氨酸脱氨酶 carbobenzyloxy-L-leucine degr 3.85 0.01
2-甲基戊二酸 2-methylglutaric acid 2.70 0.03
甲酮戊二酸 alpha-ketoglutaric acid 1.77 0.01
3-苯基乳酸 3-phenyllactic acid 1.75 0.05
半乳糖 xylose 2.31 0.01
乳清酸 orotic acid 3.72 0.00
肉豆蔻酸 myristic acid 1.03 0.00
6-磷酸葡萄糖 glucose-6-phosphate 3.63 0.03
龙胆糖 gentiobiose 4.24 0.00
I vs H 萘 naphthalene 3.47 0.00
3-羟基缬氨酸 3-hydroxynorvaline 1.39 0.05
癸酸 capric acid 1.52 0.01
薄荷酮 menthone 1.14 0.00
乳清酸 orotic acid 3.11 0.00
金合欢醛 farnesal 1.61 0.00
葡萄糖 glucose 1.40 0.00
6-磷酸果糖 fructose-6-phosphate 1.56 0.00
松二糖 turanose 2.98 0.00
龙胆糖 gentiobiose 2.79 0.00
O vs N 甘氨酸 glycine 1.15 0.00
草酸 oxalic acid 2.67 0.00
萘 naphthalene 2.99 0.00
2-丁炔-1,4-二醇 2-butyne-1,4-diol 1.98 0.00
环己烷-1,2-二醇 cyclohexane-1,2-diol 3.14 0.00
二氢香芹醇 (-)-dihydrocarveol 2.19 0.00
1,4-环己二酮 1,4-cyclohexanedione 1.92 0.05
1-茚醇 1-indanol 2.93 0.00
2-甲基戊二酸 2-methylglutaric acid 2.79 0.00
癸酸 capric acid 2.08 0.01
N-乙基马来酰胺酸 N-ethylmaleamic acid 1.86 0.01
天冬氨酸 aspartic acid 1.49 0.03
薄荷酮 menthone 3.16 0.00
阿洛糖 allose 1.30 0.00
乳清酸 orotic acid 2.67 0.00
反式-4-羟基-L-脯氨酸 trans-4-hydroxy-L-proline 1.17 0.03
金合欢醛 farnesal 2.72 0.00
葡萄糖 glucose 2.53 0.00
异肌醇 allo-inositol 1.25 0.03
槐糖 sophorose 2.18 0.03
龙胆糖 gentiobiose 2.39 0.00
半乳糖 melibiose 1.16 0.00
6-磷酸蔗糖 sucrose-6-phosphate 1.58 0.00
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不同加工方式对牛肉干产品风味物质的影响
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王美仁 1 , 王柏辉 2 , 杨帆 1, * , 刘学勤 1 , 苏春霞 1 , 段晓蓉 1
食品安全质量检测学报 | 本期专题：食品加工工艺优化及应用研究 2025,16(2): 127-137
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食品安全质量检测学报 | 本期专题：食品加工工艺优化及应用研究 2025, 16(2): 127-137
不同加工方式对牛肉干产品风味物质的影响
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王美仁1 , 王柏辉2, 杨帆1, * , 刘学勤1, 苏春霞1, 段晓蓉1
作者信息
  • 1.鄂尔多斯生态环境职业学院, 鄂尔多斯 017010
  • 2.鄂尔多斯市检验检测中心, 鄂尔多斯 017010

    • 王美仁(1985—), 女, 硕士, 讲师, 实验师, 主要研究方向为食品加工与安全。E-mail:

通讯作者:

* 杨帆(1971—), 男, 硕士, 教授, 主要研究方向为食品加工与安全方向。E-mail:
Effects of different processing methods on flavor substances of beef jerky products
Mei-Ren WANG1 , Bo-Hui WANG2, Fan YANG1, * , Xue-Qin LIU1, Chun-Xia SU1, Xiao-Rong DUAN1
Affiliations
  • 1. Ordos VocationaL College of Eco-environment, Ordos 017010, China
  • 2. Ordos City Inspection and Testing Center, Ordos 017010, China
出版时间: 2025-01-25 doi: 10.19812/j.cnki.jfsq11-5956/ts.20241021003
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摘要
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目的 研究不同加工处理方式牛肉干产品挥发性风味物质的特征。方法 将牛肉用不同方式(腌制、加菌腌制、风干等15种)进行加工, 对其进行顶空固相微萃取结合全二维气相色谱-飞行时间质谱分析。结果 检测15种样品中的差异代谢物质, 其中添加木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制的过程中, 显著性差异代谢物质包含糖、酸、酯类等共12种; 以风干方式加工的腌制牛肉和加菌腌制牛肉, 在风干后产生麦芽糖、癸酸和乳清酸等8种极显著差异代谢物; 以正常烘烤方式加工的腌制牛肉比真空烘烤加工的腌制牛肉多检出包含萘、3-羟基-DL-正缬氨酸、左旋薄荷酮、正癸酸、D-松二糖、肉豆蔻酸在内的11种差异代谢物, 并且真空烘烤参与的代谢通路有柠檬酸循环、丙氨酸、天冬氨酸和谷氨酸代谢等, 烘烤参与的代谢通路为氨基糖和核苷酸糖代谢。结论 本研究分析了不同加工处理方式的牛肉干产品挥发性风味物质的变化规律, 为提升风干牛肉的品质和口感提供了方法指导。

差异代谢物  /  气相色谱-飞行时间质谱法  /  代谢组学  /  风干  /  烘烤  /  真空烘烤  /  牛肉干

Objective To study the characteristics of volatile flavor substances in beef jerky products processed by different processing methods. Methods Beef was processed by different processing methods (15 kinds of curing, bacterial curing, air-drying, etc.), and analyzed by headspace solid-phase microextraction combined with comprehensive two-dimensional gas chromatography-time of flight mass spectrometry. Results The differential metabolites in 15 kinds of samples were detected. Among them, during the pickling process of Staphylococcus xylosus, Lactobacillus plantarum and Pediococcus pentosaceus, the significant differential metabolites included 12 kinds of sugars, acids and esters. The cured beef processed by air-drying method and the cured beef with bacteria produced 8 kinds of extremely significant different metabolites such as maltose, decanoic acid and orotic acid after air-drying. More than 11 kinds of differential metabolites including naphthalene, 3-hydroxy-DL-norvaline, L-menthone, n-decanoic acid, D-pinobiose and myristic acid were detected in the cured beef processed by normal baking than in the cured beef processed by vacuum baking. The metabolic pathways involved in vacuum baking were citric acid cycle, alanine, aspartic acid and glutamic acid metabolism, and the metabolic pathways involved in baking were amino sugar and nucleotide sugar metabolism. Conclusion In this study, the changes of volatile flavor substances in beef jerky products with different processing methods are analyzed, which provide methodological guidance for improving the quality and taste of air-drying beef.

differential metabolites  /  gas chromatography-time-of-flight mass spectrometry  /  metabolomics  /  air-dry  /  baking  /  vacuum baking  /  beef jerky
王美仁, 王柏辉, 杨帆, 刘学勤, 苏春霞, 段晓蓉. 不同加工方式对牛肉干产品风味物质的影响. 食品安全质量检测学报, 2025 , 16 (2) : 127 -137 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241021003
Mei-Ren WANG, Bo-Hui WANG, Fan YANG, Xue-Qin LIU, Chun-Xia SU, Xiao-Rong DUAN. Effects of different processing methods on flavor substances of beef jerky products[J]. Journal of Food Safety & Quality, 2025 , 16 (2) : 127 -137 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20241021003
正文
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0 引言
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牛肉是我国主要的肉类之一, 仅次于猪肉, 是重要的蛋白质来源。它含有高质量的蛋白质和低脂肪, 尤其是不饱和脂肪酸, 有助于降低胆固醇和预防心血管疾病。牛肉还含有铁、锌、磷等矿物质[1]。营养学家视牛肉为“肉中之王”, 其营养成分丰富且易吸收。牛肉含蛋白质、矿物质等, 有助于生长发育、增强免疫力、调理脾胃、强健骨骼。适量食用牛肉对各年龄段人群健康有益。干腌牛肉以独特风味和品质特征著称[2]。风干牛肉中含多种风味物质, 如醛类、酮类、酯类等挥发性化合物, 以及氨基酸、肽类等非挥发性物质[3]。研究发现不同加工工艺会影响风干牛肉风味。传统自然风干能保留原始风味, 但时间长、受环境影响大; 低温烘烤等现代工艺可缩短时间、减少微生物污染, 且能产生独特烤香味。原料肉的品质、新鲜度以及加工过程中的卫生条件等都会影响最终的风味表现[4]。乳酸菌、酵母菌、芽孢杆菌作为常见的食用益生菌, 研究表明可明显提升畜禽肉质的色泽、降低滴水损失及pH等, 也使蛋白质、脂肪酸以及各种氨基酸含量等显著提高进一步改善肉的风味, 提升肉的等级[5-6]。风干牛肉是多地传统美食, 研究其风味有助于传承制作技艺和文化内涵, 结合益生菌发酵技术, 可以拓宽风干牛肉的应用市场, 推动传统美食现代化发展[7]。在竞争激烈的休闲食品市场, 独特风味是吸引消费者的关键。深入研究风味, 可打造差异化产品, 提高产品附加值和竞争力, 助力企业占据市场份额。
目前关于风干牛肉的研究已经取得了一定的成果, 但对于不同部位及不同加工工艺的风干牛肉风味研究尚显不足, 特别是气味和滋味方面的研究较少[8]。此外, 随着消费者对食品品质和口感的要求不断提高, 如何进一步提升风干牛肉的品质和口感, 也是未来研究的重要方向[9]。
本研究以内蒙古自治区鄂尔多斯市鄂托克旗查布其农牧民专业合作社的黄牛为原料, 以不同菌液添加的加工方式进行牛肉风干, 采用定量描述分析和顶空固相微萃取结合全二维气相色谱-飞行时间质谱(comprehensive two-dimensional gas chromatography-time-of-flight mass spectrometry, GC×GC-TOF/MS)技术对不同加工处理方式的牛肉干产品挥发性风味物质差异特征和挥发性组分差异特征进行解析, 分析测定风干和贮藏过程中理化指标、微生物指标以及挥发性风味物质的变化规律, 深入研究风味前体物质与挥发性风味物质之间的相关性, 为传统风干牛肉的标准化生产、品质控制以及技术改进奠定一定的理论依据。
1 材料与方法
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1.1 材料与仪器
1.1.1 牛肉
牛肉来自内蒙古自治区鄂尔多斯市鄂托克旗查布其农牧民专业合作社同一饲养条件的黄牛。采样时调研肉品成熟度是否一致, 根据实验采样需求, 取样时间为同一生长期, 体脂率相近的黄牛。采集的样品分别装入采样袋, 贮存在-80 ℃冰箱备用[10]。
1.1.2 菌株
木糖葡萄球菌(Staphylococcus xylosus)、植物乳杆菌(Lactobacillus plantarum)、戊糖片球菌(Pediococcus pentosaceus)均保藏于内蒙古农业大学食品科学与工程学院食品实验室。
1.1.3 主要仪器设备与试剂
18L真空滚揉机(广陵食品机械有限公司); HHS-11-2恒温水浴锅(上海博讯实业有限公司); KDC-140HR高速冷冻离心机(安徽中科中佳科学仪器有限公司); PB-400pH计(北京赛多利斯仪器系统有限公司); DHP-9802电热恒温干燥箱(上海一恒科学仪器有限公司); 7890A气相色谱仪(美国AgiLent公司); PEGASUS HT质谱仪(美国LECO公司); Heraeus Fresco17离心机(美国Thermo Fisher Scientific公司); BSA124S-CW分析天平 (精度0.0001 g, 德国Sartorius公司); JXFSTPRP-24研磨仪(上海净信科技有限公司); LNG-T98真空干燥仪(太仓市华美生化仪器厂)。
核糖醇标准品(纯度≥99%)、C7~C30直链正构烷烃、乙醇(色谱纯)(美国Sigma-ALdrich公司); 甲氧胺盐(分析纯, 日本TCI公司); NaCl(分析纯, 中国医药集团化学试剂有限公司)。
1.2 实验方法
1.2.1 牛肉加工工艺
牛肉去筋, 切条加食盐、糖、五香粉、料酒、红酱油、味精、白胡椒粉、菌液、纯净水放入滚揉机滚揉。滚揉完成后, 放入盆中。选择牛肉条长厚的肉条挂到风干机中, 依据各组时间、温度风干后烘烤。
原料肉预处理: 选用黄牛后腿肉, 剔除牛肉表面筋膜等杂物。顺着肌纤维的方向将牛肉分割成约长15 cm, 宽4 cm, 厚度3 cm的肉条。
静置腌制: 添加腌制剂, 4 ℃腌制12.0 h。
真空滚揉腌制: 添加腌制剂, 0.04 MPa条件下真空滚揉4.0 h。
低温风干: 将腌制好的牛肉运入风干室, 挂肉要注意肉条之间的距离, 保持空气流通, 确保室内温度达到设定的范围[11]。
高温烘干: 将风干肉置于鼓风干燥箱中进行烘干。
腌制剂配比(每kg肉): 食盐1.5%、糖0.2%、五香粉0.2%、料酒3%、红酱油0.5%、味精0.4%、白胡椒粉0.1%、菌液2%、水10%。
实验设置自然组: 30 h 30 ℃、124 h 30 ℃、36 h 36 ℃; 加菌组(肉:纯净水=10:1, m:V): 24 h 30 ℃、30 h 30 ℃, 其中A组: 鲜牛肉; B组: 未加菌腌制牛肉; C组: 添加木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制的牛肉; D组: B组经风干工艺加工的产品; E组: C组经风干工艺加工的产品; F组: B组经蒸煮、烘烤工艺加工的产品; G组: C组经蒸煮、烘烤工艺加工的产品; H组: B组经烘烤工艺加工的产品; I组: C组经烘烤工艺加工的产品; J组: B组经真空烘烤工艺加工的产品; K组: C组经真空烘烤工艺加工的产品; L组: B组经真空烘烤和冻干工艺加工的产品; M组: C组经真空烘烤和冻干工艺加工的产品; N组: B组经高压蒸煮工艺加工的产品; O组: C组经高压蒸煮工艺加工的产品。
1.2.2 GC×GC-TOF/MS实验方法
取样本(50±1) mg于2 mL离心管中, 加入1000 μL预冷提取液(甲醇:氯仿=3:1, V:V), 含核糖醇, 涡旋30 s; 加入钢珠, 40 Hz研磨仪处理4 min, 冰水浴超声5 min(重复3次); 将样本4 ℃离心, 12000 r/min离心15 min小心移取300 μL上清液于1.5 mL离心管中, 每个样本各取100 μL混合成质控(quality control, QC)样本; 在真空浓缩器中干燥提取物; 向干燥后的代谢物加入40 μL甲氧胺盐试剂(甲氧胺盐酸盐溶于20 mg/mL吡啶), 轻轻混匀后, 放入烘箱中80 ℃孵育30 min; 向每个样品中加入50 μL N,O-双(三甲基硅烷基)三氟乙酰胺[N,O-bis(trimethylsilyl) trifluoroacetamide, BSTFA](含有1%三甲基氯化亚砜, V:V), 将混合物70 ℃孵育1.5 h; 冷却至室温, 向混合的样本中加入5 μL脂肪酸甲酯(溶于氯仿); 随机顺序上机检测[12-13]。
1.2.3 GC×GC-TOF/MS分析方法
使用AgiLent7890气相色谱仪结合飞行时间质谱仪进行GC×GC-TOF/MS分析。该系统采用DB-5MS毛细管柱。最大以无分流模式注入等分试样。使用氦气作为载气, 前入口吹扫流量为3 mL/min, 通过柱的气体流速为1 mL/min。初始温度保持在50 ℃/min, 以10 ℃/min的速率升高到310 ℃, 然后在310 ℃保持8 min。注入、传输线和离子源的温度分别为280、280和250 ℃。在电子撞击模式下能量为-70 V。在全扫描模式下, 以每秒12.5光谱的速率获取质荷比(m/z)范围为50至500的质谱数据, 溶剂延迟6.4 min[14]。
1.3 数据处理
使用ChromaTOF软件(V4.3x, LECO)对质谱数据进行峰提取、基线矫正、解卷积、峰积分、峰对齐等分析。对物质定性工作中, 使用了LECO-FiehnRtx5数据库, 包括质谱匹配及保留时间指数匹配。最后, 将QC样本中检出率50%以下或相对标准偏差(relative standard deviation, RSD)>30%的峰去除[15]。
2 结果与分析
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2.1 质量控制
2.1.1 过程质控
当样本量过多、检测时间过长时, 需要在检测过程中实时监控仪器稳定性, 及时发现异常, 尽早将问题排除, 以保证最终采集数据的质量。由表1可知, 内标在QC样品中峰面积RSDs=5.56%≤30%, 说明仪器数据采集稳定。
在QC过程中, 总离子色谱(total ion chromatogram, TIC)图可以用来确认样品的稳定性和仪器的性能, 可以更好地理解和分析样品中的复杂化合物。通过对空白样品的检测可以考察检测过程中物质残留情况。由图1可知, QC样本TIC出峰保留时间和峰面积均重叠适宜, 说明仪器稳定。通过观察TIC中的背景噪音和意外出现的峰, 可以识别和排除可能的污染物或仪器故障。由图2可知, 空白样品中无显著峰检出, 说明物质残留控制适当, 不存在样品间的交叉污染[16]。
2.1.2 数据质控
QC样本在物质提取、检测分析的过程中会有系统误差, 导致QC样品间存在差异。这个差异越小说明整个方法稳定性越好、数据质量越高。由图3可知, 样本几乎全部分布在±2 std范围内, 因此本研究的数据更加可靠、准确[17]。
2.2 统计学分析
2.2.1 主成分分析
由图4a可知, 各组之间的样本呈现分离趋势, 尤其不同工艺对样本影响较大, 使得样本分离明显。由图4b可知, 第一成分贡献率为21.1%, 第二成分贡献率为9.5%, 结合两幅图可以更加直观地看出各样本分散情况, 从二维图中看出整体分离效果较好, 说明不同组别牛肉干风味物质存在差异, 但有少部分重叠[18], 而PC1>PC2, 以横坐标为主说明横坐标距离越大, 样本差异性越大[2,19]。
2.2.2 正交偏最小二乘法-判别分析
采用的统计方法对结果进行分析。通过正交偏最小二乘法-判别分析(orthogonal projections to latent structures discriminant analysis, OPLS-DA), 能够获取更加可靠的代谢物的组间差异与实验组的相关程度信息。OPLS-DA结果见图5, 由图5可知, 15组的样品未交叉重叠, 说明代谢信息不同; 不同处理方式下的样本横向距离较远, 说明组间差异明显; 大部分样本组内纵向距离较近, 重复性好。各组得分散点图下方对应置换检验结果, 各组别R2均接近于1, Q2趋向于0, 表明OPLS-DA模型稳定可靠, 没有出现过拟合现象, 组间分离明显, 无交叉重合[20], 模型合理且鉴定的代谢物可用于下一步分析。
2.3 差异代谢物分析
2.3.1 差异代谢物聚类分析
为对比菌种、烘烤、冻干等方式对牛肉干风味物质的影响, 对以下10组进行差异代谢物聚类分析, 结果见图6, 由图6a可知, 经过加入木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制过的牛肉(C组)中, 2-脱氧-D-葡萄糖、环氧缩酮、6-磷酸蔗糖含量增加, 癸酸、乳清酸、葡萄糖酸内酯、1,2-环己二酮等物质含量降低。由图6b可知, 牛肉经过风干后癸酸、麦芽糖、乳清酸等物质含量增加, 同时比未加菌种的D组高, 可能是加入菌种后产生的相关物质增加了牛肉干的风味[21-22]。通过对比图6c与6d可知, 经过真空烘烤后, 隔绝了氧气, 限制了各菌种对物质的产生, 降低了J组与K组整体代谢物的数量。由图6e可以得到: 经过高压蒸煮后两组产生了一些新物质如反式-4-羟基-L-脯氨酸、阿洛糖、胞苷酸、草酸等物质, 经过加菌腌制过的O组新产生天冬氨酸、甘氨酸等物质。
2.3.2 差异代谢物的筛选
基于OPLS-DA模型, 以变量重要性投影(variable importance for the projection, VIP)>1、P<0.05为标准进行筛选, 对比不同组别差异代谢物, 结果见表2。通过筛选后对比C组与B组可知, 腌制牛肉时加入木糖葡萄球菌、植物乳杆菌和戊糖片球菌后, 有效增加了牛肉的风味物质, 其中具有显著性差异代谢物有糖类、酯类、酸类共12种, 改善了牛肉的品质[23-24]。C组与B组经过风干后得到E组与D组, 其中如麦芽糖、癸酸、乳清酸、戊糖、6-磷酸蔗糖等5种物质经过干制后仍为极显著差异代谢物, 说明这几种物质在牛肉干的制作过程中起到重要作用。孔琪[25]将鼠李糖乳杆菌LR1、植物乳杆菌LP5、乳酸片球菌PA45发酵液加入牛肉经过干制后对牛肉干的品质进行分析得出结论: 乳酸菌发酵液能够优化调理牛肉的感官品质。将C组与B组经过烘烤后得到I组与H组, 由表2可知, 经过烘烤后产生了新的物质3-羟基缬氨酸、门冬酰胺、呋喃糖等。从整体上看, D、E组特有的差异代谢物为5-氨基戊酸内酰胺、酵母醇总计2种, J、K组特有的差异代谢物为苄氧羰基-L-亮氨酸脱氨酶、2-甲基戊二酸、α-酮戊二酸、3-苯基乳酸总计4种, H、I组特有的差异代谢物为3-羟基缬氨酸、松二糖2种, O、N组特有的差异代谢物为甘氨酸、草酸、2-丁炔-1,4-二醇、环己烷-1,2-二醇、二氢香芹醇、1,4-环己二酮、1-茚醇、N-乙基马来酰胺酸、天冬氨酸、阿洛糖、反式-4-羟基-L-脯氨酸、异肌醇、槐糖总计13种, 综上所述O、N组高压蒸煮的处理方式使得牛肉的风味物质增多[26]。
2.3.3 差异代谢物的雷达图分析
通过雷达图对比各组差异代谢物含量趋势变化, 如图7所示。图中用红色字体表示, 每一条网格线表示一个差异倍数, 紫色阴影由每个物质的差异倍数连线组成。
由图7可知, B、C组中麦芽糖、乳清酸的得分最高, 风味贡献最大, 为牛肉提供了牛奶清香和甜味。周亚军等[27]以牛臀肉为原料, 接种木糖葡萄球菌和清酒乳杆菌制作发酵牛肉干, 对牛肉干发酵前后蛋白质含量、质构、质构、色泽游离氨基酸、脂肪酸组成和挥发性风味物质对比分析, 结果表明: 接种发酵剂的牛肉干pH、硬度均显著降低, 经过发酵后, 挥发性物质增加5种, 同时提高了牛肉干中醛、醇、酚、酯、酸等物质的相对含量, 使得产品营养与风味显著提高。D、E组风干牛肉的风味物质种类有所下降, 可能的原因是在普通风干过程中流失, 只有5-氨基戊酸内酰胺的评分最高, 其他物质评分接近。
经过真空烘烤, K、J组各风味物质的贡献率都有所增加, 其中6-磷酸葡萄糖的评分较低, 其他物质对风味的贡献均比较大。采用真空的方式进行烘烤, 有效防止牛肉的氧化, 尽可能地保留了牛肉原始风味。对比H、I组可得, 在正常条件下烘烤, 各物质对风味的贡献相近, 同时也说明各物质产生的风味较平和, 使得牛肉干风味平衡。经过高压蒸煮后, 破坏了大部分风味物质, 降低了风味物质的评分, 导致牛肉干的风味不明显, 如图7所示, O、N组可能无法使人感知。
2.3.4 差异代谢物的相关性分析
进行差异代谢物相关性分析有利于进一步了解生物状态变化过程中代谢物之间的相互调节关系。为了探究各代谢物之间的相关性, 通过对比B与C、D与E、J与K、H与I组间代谢物, 探究各物质之间的相关程度, 结果见图8。由图8可知, C vs B组中代谢物间均具有显著性, 相关性系数的绝对值接近1, 甚至有24种相关性系数等于1, 可能是接入菌种后产生各种物质, 提高了牛肉的风味[28]。从E vs D组中可以看出, 牛肉经过风干后风味物质有所减少, 但各物质之间相关性系数也较高, 也具有显著性。从整体看, 随着处理温度的升高, 呈负相关的物质占比逐渐增多, 部分物质失去了显著性。
2.3.5 差异代谢物的KEGG富集分析
根据京都基因和基因组百科全书(kyoto encyclopedia of genes and genomes, KEGG)通路注释的结果对B、C、D、E、H、I、J、K组别的样本进行分类, 结果见图9。
如图9所示, C vs B组中新陈代谢途径差异代谢物的数量最高占85.71%, 其次为戊糖磷酸途径、碳水化合物代谢、ABC转运体, 各占28.57%, 其他均为14.29%。E vs D组中主要代谢通路为靛酚和蔗糖代谢、半乳糖代谢、碳水化合物代谢、辅因子的生物合成、代谢途径、类固醇生物合成、脂肪酸的生物合成、ABC转运蛋白、嘧啶代谢, 其中ABC转运蛋白占比最高为33.33%,其余均为16.67%,与C vs B组相比增加了类固醇的生物合成。I vs H组中代谢通路与C vs B组相比种类明显增加, 产生了腺苷酸活化蛋白激酶(adenosine monophosphate-activated protein kinase, AMPK)信号通路、胰岛素抵抗等新的代谢通路, 各通路差异代谢物的数量占比均匀[29]。而K vs J组与I vs H组相比, 代谢通路又有所增加, 可能原因是在接触空气后经过高温产生的, K vs J组中代谢通路与前3组大不相同, 各代谢通路差异代谢物占比较I vs H组有所增加。
2.3.6 差异代谢物的代谢通路分析
通过对差异代谢物所在通路的富集分析, 可以对通路进行进一步的筛选, 找到与代谢物差异相关性最高的关键通路, 代谢通路分析的结果以气泡图展示, 结果见图10。
由图10可知, C vs B组中戊糖磷酸途径的显著性最高但是其影响因子几乎为0, 该组中只有半乳糖代谢通路的影响因子和显著性均较高, 可能是因为加入木糖葡萄球菌、植物乳杆菌和戊糖片球菌后消耗了乳糖或半乳糖, 使得半乳糖代谢通路上的物质得到富集并具有显著性。经过干制后得到E vs D组, 半乳糖代谢通路显著性有所提高, 新得到类固醇生物合成通路, 该通路影响因子和显著性均较低[30]。相比前两组, K vs J组中TCA循环、丙氨酸、天冬氨酸和谷氨酸代谢、嘧啶代谢显著性高, 前两条通路的影响因子也较高。经过K vs J组真空烘烤后, 代谢通路数量增加, 各通路的影响因子也比其他组高。
3 结论
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本研究使用GC×GC-TOF/MS技术探究不同加工处理方式风干牛肉中风味物质的变化规律。结果显示, 加木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制, 显著性差异代谢物有糖、酯、酸类等共12种, 在牛肉风味形成过程中发挥重要作用, 通过KEGG通路富集分析发现代谢途径、戊糖磷酸途径、碳代谢、ABC转运蛋白是主要的代谢途径, 且半乳糖代谢通路上的物质得到富集并具有显著性; 将风干牛肉与未风干的牛肉进行对比, 发现风干牛肉产生麦芽糖、癸酸和乳清酸等8种极显著差异代谢物, 通过雷达图和相关性分析充分表明风味物质数量减少; 将烘烤和真空烘烤的牛肉进行比较, 真空烘烤的牛肉整体风味化合物数量降低, 6-磷酸葡萄糖对风味的贡献也较小, 通过代谢通路分析发现TCA循环、丙氨酸、天冬氨酸和谷氨酸代谢、嘧啶代谢具有显著性, 影响因子也比其他组高。相比之下, 接入木糖葡萄球菌、植物乳杆菌和戊糖片球菌腌制的风干牛肉的品质最优, 在半乳糖代谢、类固醇生物合成和嘧啶代谢中产生的麦芽糖、癸酸、乳清酸、戊糖、6-磷酸蔗糖对风干牛肉风味物质的产生奠定基础, 为标准化生产提供依据。
基金
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  • 内蒙古自治区科技创新引导奖励资金项目(NM-KJCXYD-014)
文献
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2025年第16卷第2期
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20241021003
  • 接收时间：2024-10-21
  • 首发时间：2025-07-21
  • 出版时间：2025-01-25
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  • 收稿日期：2024-10-21
基金
内蒙古自治区科技创新引导奖励资金项目(NM-KJCXYD-014)
作者信息
    1.鄂尔多斯生态环境职业学院, 鄂尔多斯 017010
    2.鄂尔多斯市检验检测中心, 鄂尔多斯 017010

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* 杨帆(1971—), 男, 硕士, 教授, 主要研究方向为食品加工与安全方向。E-mail:
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https://castjournals.cast.org.cn/joweb/spaq/CN/10.19812/j.cnki.jfsq11-5956/ts.20241021003
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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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