[关键词]
[摘要]
目的 通过超高效液相色谱串联三重四级杆飞行时间质谱法(UPLC-Triple-TOF-MS)快速分析续断Dipsaci Radix“发汗”前后的化学成分。方法 采用Agilent Eclipse XDB-C18(250 mm×4.6 mm,5.0 μm)色谱柱;流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,体积流量为0.8 mL/min,检测波长215 nm,柱温为25℃;质谱采用UPLC-Triple-TOF 5600+飞行时间液质联用仪,负离子模式下扫描检测,扫描范围m/z 100~1 500。根据高分辨质谱结合二级质谱所得的信息对续断“发汗”前后化学成分进行快速的鉴定。结果 根据化合物保留时间、质谱信息,推测了续断“发汗”前后52个共有化学成分,主要包括三萜皂苷类、环烯醚萜类、酚酸类等,同时分析了主要成分的裂解规律。并比较了续断“发汗”前后化学成分的差异,发现发汗后马钱苷酸、绿原酸、马钱苷、异绿原酸A、川续断皂苷VI等20种成分含量降低,咖啡酸、异绿原酸、异绿原酸C、大花双参苷A等成分含量在发汗后升高。结论 研究发现续断“发汗”前后化学成分种类上无明显差异,而在成分含量上存在一定差异,未“发汗”续断化学成分含量普遍高于“发汗”续断。本实验采用UPLC-Triple-TOF-MS技术分析“发汗”对续断的化学成分的影响,为探究续断“发汗”前后化学成分物质基础和进一步为续断产地加工方式的研究提供理论依据。
[Key word]
[Abstract]
Objective To establish a quick method of ultra-performance liquid chromatography-quadrupole time-of-fight mass spectrometry (UPLC-Triple-TOF/MS) for the analysis of components of crude and sweated Dipsaci Radix. Methods The separation was performed on the chromatographic column of Agilent Eclipse XDB-C18 (250 mm×4.6 mm, 5.0 μm), and the mobile phase was 0.1% formic acid solution-methanol, with a gradient elution at a flow rate of 0.8 mL/min, the detection wavelength was 215 nm, the column temperature was 25℃. UPLC-Triple-TOF 5600+ time of flight liquid and mass spectrometer was used for mass spectrometry. Electrospray ion source negative ion mode was adopted, and the scanning range was m/z 100-1 500. The components of crude and sweated Dipsaci Radix were quickly identified according to the information obtained by high-resolution mass spectrometry combined with secondary mass spectrometry. Results Fifty-two common components were identified or tentatively characterized based on the retention time and MS spectra. They were triterpenoid saponins, iridoids, phenolic acids etc. The crack rules of primary components were also analyzed. And comparing the components of crude and sweated Dipsaci Radix, it showed that the content of 20 components such as loganin acid, chlorogenic acid, loganin, isochlorogenic acid A, and asperosaponin VI was decreased after sweating, and caffeic acid, isochlorogenic acid, isochlorogenic acid C, and triplostoside A was increased. Conclusion The types of components of crude and sweated Dipsaci Radix are identical, but there are differences in the content of the components. The content of the components of crude are higher than the sweated Dipsaci Radix. UPLC-Triple-TOF-MS technology was used to analyze the influence of "sweating" on the chemical composition of the Dipsaci Radix, so as to provide a theoretical basis for the study of the chemical constituents of sweated Dipsaci Radix and further research on the origin processing of Dipsaci Radix.
[中图分类号]
R283.1
[基金项目]
国家重点研发计划:中药饮片质量识别关键技术研究(2018YFC1707001);国家中药标准化项目(ZYBZH-H-ZY-45);中华中医药学会青年人才托举工程项目(QNRC2-C12);浙江省药监局委托产地加工项目(ZJ-197505-1);浙江中医药大学校级重点项目(2019ZZ10);象山县科技计划项目