基于非靶向代谢组学的外源合成冰乙酸(乙醇发酵法)配制食醋的鉴别及特征标志物分析

Identification and characteristic biomarker analysis of vinegar prepared with exogenous synthetic glacial acetic acid (ethanol fermentation method) based on untargeted metabolomics

中文关键词:  醋  乙醇发酵法  冰乙酸  超高效液相色谱-四极杆-静电场轨道离子阱质谱  代谢组学  鉴别

英文关键词:vinegar  ethanol fermentation method  glacial acetic acid  ultra performance liquid chromatography-quadrupole-Orbitrap tandem mass spectrometry  metabolomics  identification

中文摘要:

      目的 建立基于非靶向代谢组学技术方法, 分析酿造食醋中勾兑冰乙酸(乙醇发酵法)后的代谢物差异, 筛选特征标志物用于掺假鉴别。方法 以宁化府工艺老陈醋为研究对象, 采用超高效液相色谱-四极杆-静电场轨道离子阱质谱法(ultra performance liquid chromatography-quadrupole-Orbitrap tandem mass spectrometry, UPLC-Q-Orbitrap MS/MS)对不同勾兑程度食醋样品(酸度6.0%~8.0%)的代谢物成分进行采集, 结合主成分分析和偏最小二乘判别分析等多元统计分析方法筛选并鉴定差异代谢产物。结果 共筛选出5个具有显著性差异的标志物, 包括油酸酰胺、N-苄基-亚油酰胺、柠檬酸三乙酯、己二酸二丁酯和单棕榈酸甘油。当油酸酰胺含量明显低于479.99 μg/kg以及N-苄基-亚油酰胺、柠檬酸三乙酯、己二酸二丁酯和单棕榈酸甘油含量高于传统酿造食醋水平(分别为0.92、0.89、10.82 μg/kg和56.56 mg/kg)时, 可有效判别食醋是否掺假。油酸酰胺含量随冰乙酸浓度升高呈明显下降趋势, 在酸度为8.0%的样品中含量为39.40 μg/kg。N-苄基-亚油酰胺(9.50 μg/kg)在酸度为7.0%时达到峰值, 柠檬酸三乙酯(4.22 μg/kg)、单棕榈酸甘油(129.26 mg/kg)与己二酸二丁酯(27.66 μg/kg)含量在酸度8.0%样品中分别达到传统食醋的4.75、2.29和2.56倍。结论 本研究建立的基于UPLC-Q-Orbitrap MS/MS的非靶向代谢组学方法, 筛选出的显著性差异标志物, 为食醋真实性鉴定提供了一种新型的技术手段, 保障食醋产品质量及保护消费者健康权益。

英文摘要:

      Objective To establish a technical method based on untargeted metabolomics, analyze the differences in metabolites after blending glacial acetic acid (ethanol fermentation method) in brewed vinegar, and screen characteristic markers for adulteration identification. Methods Taking Ninghuafu traditional aged vinegar as the research object, ultra performance liquid chromatography-quadrupole-Orbitrap tandem mass spectrometry (UPLC-Q-Orbitrap MS/MS) was used to collect metabolite components of vinegar samples with different dilution degrees (acidity ranging from 6.0%–8.0%). Combined with multivariate statistical analysis methods such as principal component analysis and partial least squares discriminant analysis, differential metabolites were screened and identified. Results A total of 5 markers with significantly differences were identified, including oleamide, N-benzyl-linoleamide, triethylcitrate, dibutyladipate and monopalmitin. When the content of oleamide was significantly lower than 479.99 μg/kg, and the content of N-benzyl-linoleamide, triethylcitrate, dibutyladipate and monopalmitin were higher than the levels in traditional brewed vinegar (0.92, 0.89, 10.82 μg/kg and 56.56 mg/kg, respectively), these compounds could effectively distinguish whether the vinegar had been adulterated. The content of oleamide showed a significant downward trend with increasing glacial acetic acid concentration, with a content of 39.40 μg/kg in samples with an acidity of 8.0%. N-benzyl-linoleamide (9.50 μg/kg) reached peaks in the sample groups 7.0% acidity. Triethylcitrate (4.22 μg/kg), monopalmitin (129.26 mg/kg) and dibutyladipate (27.66 μg/kg) in the 8.0% acidity sample reached 4.75, 2.29 and 2.56 times that of traditional vinegar. Conclusion The untargeted metabolomics method based on UPLC-Q-Orbitrap MS/MS established in this study screens out significantly differential biomarkers, which provides a novel technical approach for the authenticity identification of vinegar, and guarantees the quality of vinegar products as well as protects the health rights and interests of consumers.