UPLC-Q-TOF-MS结合分子量印迹和质量亏损过滤策略快速筛选并鉴定血竭中黄酮类化学成分

Rapid flavonoid-focused sub-chemome characterization of Draconis Sanguis using UPLC-Q-TOF-MS in combination with molecular weight imprinting and mass defect filtering

血竭为名贵活血化瘀中药,现有研究表明其药效成分群为黄酮类化合物。血竭中黄酮类化合物化学结构的多样性为其化学成分轮廓深入表征带来了巨大的挑战。为了系统表征血竭化学成分轮廓,该研究首先采集化学成分质谱信息,进一步利用分子量印迹(molecular weight imprinting,MWI)以及质量亏损过滤(mass defect filtering,MDF)方法处理高分辨数据集,实现血竭中黄酮类化合物的快速鉴定。采用UPLC-Q-TOF-MS采集正离子模式下m/z 100~1000质谱数据;根据前期文献报道中的血竭黄酮类化合物,建立[M+H]+质荷比的数据库,以±10×10^(-3)为误差范围构建分子量印迹,快速检索已知黄酮类化合物,并结合MS2数据确证化学结构。通过已知血竭黄酮类化合物建立MDF的“五点筛选框”,获取可能的目标成分,进一步结合诊断离子(DFI)、中性丢失(NL)和质谱裂解规律,推断化学结构。最终从血竭提取物中初步鉴定了70个黄酮类化合物,包括黄烷氧化同系物类5个、黄烷类12个、二氢查尔酮类1个、黄酮二聚体类49个、三聚体类1个以及其他类型2个。该研究阐明了血竭中黄酮类化学成分组成,同时也表明高分辨质谱技术结合MWI与MDF等数据后处理方法可以实现中药化学成分组成的快速表征。

Draconis Sanguis is a precious Chinese medicinal material for activating blood and resolving stasis, and its effective components are flavonoids. However, the structural diversity of flavonoids in Draconis Sanguis brings great challenges to the in-depth chara-cterization of its chemical composition profiles. To clarify the substance basis of Draconis Sanguis, ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) was used in this study to acquire MS data of Draconis Sanguis. The molecular weight imprinting(MWI) and mass defect filtering(MDF) were developed for rapid screening of flavonoids in Draconis Sanguis. Full-scan MS and MS~2 were recorded within the mass range m/z 100-1 000 in positive ion mode. Accor-ding to previous literature, MWI was employed to hunt for reported flavonoids in Draconis Sanguis, and the mass tolerance range of [M+H]+was set as ±10×10^(-3). A five-point MDF screening frame was further constructed to narrow the screening range of flavonoids from Draconis Sanguis. Combined with diagnostic fragment ions(DFI) and neutral loss(NL) as well as mass fragmentation pathways, 70 compounds were preliminarily identified from the extract of Draconis Sanguis, including 5 flavan oxidized congeners, 12 flavans, 1 dihydrochalcones, 49 flavonoids dimers, 1 flavonoids trimer and 2 flavonoid derivatives. This study clarified the chemical composition of flavonoids in Draconis Sanguis. Moreover, it also showed that high-resolution MS combined with data post-processing methods such as MWI and MDF could achieve rapid characterization of the chemical composition in Chinese medicinal materials.