UHPLC-Oribtrap/HRMS技术分析干姜中非挥发性化学成分在炮制前后的变化
Study on Variation of Non-Volatile Compounds of Raw Ginger after Processing by UHPLC-Orbitrap/HRMS
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摘要: 目的 基于超高效液相色谱-静电场轨道阱高分辨质谱联用(UHPLC-Oribtrap/HRMS)技术对干姜炮制前后非挥发性化学成分的变化进行分析。方法 通过80%甲醇提取和UHPLC-Oribtrap/HRMS数据采集，结合保留时间，精确分子质量及二级质谱碎裂规律对干姜特征性成分进行鉴定。利用多变量分析对干姜炮制前后的差异化学成分进行筛选并评价其与功效相关的药理活性。结果 共鉴定72个特征化合物。主成分分析（PCA）及正交偏最小二乘法-判别分析(OPLS-DA)表明干姜炮制前后化学成分谱存在显著差异，结合模型的变量重要因子和非参数检验，筛选出11个差异物质，分别鉴定为：6-姜烯酚(6-Shogaol)，8-姜烯酚(8-Shogaol)，10-姜烯酚(10-Shogaol)，12-姜烯酚(12-Shogaol)，6-姜酚(6-Gingerol)，10-姜酚(10-Gingerol)，姜酮酚(6-Paradol)，1-去氢-6-姜二酮(6-Dehydrogingerdione)，1-去氢-8-姜二酮(8-Dehydrogingerdione)，甲基乙酰氧基-6-姜二醇(Methylacetoxy-6-gingerdiol)，7-(4-羟基苯基)-1-苯基-4-庚烯-3-酮7-(4-Hydroxyphenyl)-1-phenylhept-4-en-3-one。其中姜烯酚类化合物显示出较好的抗炎活性。结论 为合理科学解释干姜和炮姜的功效差异及炮制原理提供一定的依据。Abstract: OBJECTIVE To study the different non-volatile compounds between raw ginger and processed ginger by UHPLC-Orbitrap/HRMS. METHEODS Raw ginger and processed ginger were extracted by 80% methanol， respectively. The characteristic compounds were identified by retention time， accurate molecular weight and MS fragmentation. The different compounds were screened by multivariate analysis and their bioactivities were measured. RESULTS Seventy-two compounds were identified or tentatively characterized. The combination of principal component analysis (PCA) and orthogonal partial least squares-discriminant analysis (OPLS-DA) revealed the different chemical profiles of raw ginger and processed ginger and 11 compounds， including 6-shogaol， 8-shogaol， 10-shogaol， 12-shogaol， 6-gingerol， 10-gingerol， 6-paradol， 6-dehydrogingerdione， 8-dehydrogingerdione， methylacetoxy-6-gingerdiol and 7-(4-hydroxyphenyl)-1-phenylhept-4-en-3-one were found changed after processing. Among those compounds， shogaols showed significant anti-inflammatory activities. CONCLUSION The results provide evidence to explain the efficacy differences between raw ginger and processed ginger as well as the processing mechanism.
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