基于电子舌和UPLC-HRMS的蒙药额尔敦-乌日勒基础方“传统药味-化学成分”关联机制研究

    Correlation Mechanism Between “Traditional Flavor Profiles-Chemical Composition” of the Eerdun Wurile Basic Formula Based on Electronic Tongue and UPLC-HRMS

    • 摘要:
      目的  联合电子舌与UPLC-HRMS技术,探讨蒙药额尔敦-乌日勒基础方(Eerdun Wurile basic formula,EWB)的药味特征和化学成分,并初步建立其“药味-成分”联系。
      方法  以珍珠、甘草、土木香、木香、沉香、牛黄、荜茇、地锦草、水牛角浓缩粉、麝香为原料,依据《内蒙古蒙药制剂规范》及《内蒙古蒙药饮片炮制规范》(2020年版)制备EWB,再经回流提取、减压回收及冻干得到EWB提取物(EWB extracts,EWT);随后按分组取各药物,加入去离子水后超声震荡或煎煮处理30 min,过滤后取上清液作为电子舌待测液;利用UPLC-HRMS、NPClassifier 天然产物分类法分析EWB化学成分及分类。
      结果  电子舌结果显示,煎煮法较超声法增强甘草苦味并降低麝香苦味强度,同时弱化甘草、土木香、木香、沉香、荜茇、地锦草的鲜味(P<0.05)。煎煮法处理EWB组(SEWB)相较于煎煮法处理珍宝丸组(SEW),新增涩味,苦味、鲜味及苦味回味增加,咸味、涩味回味、鲜味回味降低(P<0.05)。而SEWB相较于超声法处理EWB组(CEWB)进一步提升苦味、涩味回味及苦味回味的强度,同样新检测到特异性涩味响应值,其咸味与鲜味响应值呈现下降趋势(P<0.05)。化学成分鉴定出EWB中含2033个特征性化合物,经NPClassifier分类主要集中于莽草酸酯-苯丙素类(27%)、萜类(22%)、生物碱(20%)三大生物合成途径。基峰色谱图标记包括甘草酸、甘草次酸、沉香四醇、荜茇宁在内共42种化合物。
      结论 本研究阐明了EWB经煎煮后呈现苦、涩、咸、鲜及回味特征,其化学成分以莽草酸酯-苯丙素类、萜类、生物碱为主,初步讨论了EWB“传统药味-化学成分”之间的可能关联。

       

      Abstract:
      OBJECTIVE To combine the electronic tongue taste system with UPLC-HRMS technology to elucidate the taste characteristics and corresponding chemical component categories of the Mongolian medicine Eerdun Wurile basic formula(EWB), and to preliminarily establish its “taste-component” relationship.
      METHODS Using Margarita, Glycyrrhizae Radix et Rhizoma, Inulae Radix, Aucklandiae Radix, Aquilariae Lignum Resinatum, Bovis Calculus, Piperis Fructus, Euphorbiae Humifusae Herba, Bubali Cornu Concentratum Pulvis and Moschus as raw materials, EWB was prepared in accordance with the Inner Mongolia Mongolian Medicine Preparation Standards and the Inner Mongolia Mongolian Medicine Decoction Piece Processing Standards(2020 Edition). Subsequently, the EWB extract(EWB extracts, EWT) was obtained through reflux extraction, vacuum recovery‌ and freeze-drying. Then, according to the grouping, each medicinal material was taken, mixed with deionized water, and subjected to ultrasonic oscillation or decoction for 30 min. After filtration, the supernatant was collected as the test solution for the electronic tongue. The chemical components and their classifications in EWB were analyzed using UPLC-HRMS and the NPClassifier natural product classification method.
      RESULTS The electronic tongue results showed that the decoction method, compared with the ultrasound method, enhanced the bitterness of Glycyrrhizae Radix et Rhizoma while reducing the bitterness intensity of Moschus, and attenuated the umami taste of Glycyrrhizae Radix et Rhizoma, Inulae Radix, Aucklandiae Radix, Aquilariae Lignum Resinatum, Piperis Fructus and Euphorbiae Humifusae Herba(P<0.05). The decoction-treated EWB group(SEWB) exhibited a newly emerged astringency, increased bitterness, umami, and bitter aftertaste, and decreased saltiness, astringent aftertaste, and umami aftertaste compared with the decoction-treated Eerdun Wurile group(SEW)(P<0.05). Furthermore, compared with the ultrasound-treated EWB group(CEWB), the SEWB showed enhanced intensities of bitterness, astringent aftertaste, and bitter aftertaste, also newly detected a specific astringency response value, while saltiness and umami response values exhibited a decreasing trend(P<0.05). Chemical composition analysis identified 2033 characteristic compounds in EWB, which were classified by NPClassifier mainly into three major biosynthetic pathways: Shikimate ester-phenylpropanoids(27%), terpenoids(22%), and alkaloids(20%). A total of 42 compounds, including glycyrrhizin, 18β-glycyrrhetinic acid, agarotetrol, and piperlonguminine, were annotated in the base peak chromatogram.
      CONCLUSION This study elucidates that after decoction, EWB exhibits bitterness, astringency, saltiness, umami, and aftertaste characteristics, with its chemical components mainly comprising shikimate ester-phenylpropanoids, terpenoids, and alkaloids. The possible correlations between the “traditional taste” and chemical composition of EWB are preliminarily discussed.

       

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