超高效液相色谱-四极杆-飞行时间质谱法分析葫芦巴成分

Chemical Constituents of Fenugreek Using UPLC-Q-TOF MS

  • 摘要: 采用超高效液相色谱-四极杆-飞行时间质谱法(UPLC-Q-TOF MS)对葫芦巴的醇提组分和除糖后的水提组分进行定性分析,并以甘草苷为内标,对鉴定出的成分进行半定量分析。采用Unitary C18色谱柱(4.6 mm×150 mm×5 μm),以乙腈-0.1%甲酸/水为流动相进行线性梯度洗脱,流速0.5 mL/min,电喷雾离子源(ESI)负离子模式检测。定性分析结果表明,葫芦巴的醇提组分和水提组分的总离子流图相似,共鉴定出36种化合物,包括12种黄酮类成分和24种皂苷类成分。实验选择了几种代表性的黄酮类和皂苷类化合物,对其结构进行详细阐述,并推断其可能的质谱裂解规律。半定量分析结果表明,两种提取物中主成分的相对含量不同,黄酮类化合物在乙醇提取物中的相对含量之和是水提物中的1.13倍,皂苷类化合物在乙醇提取物中的相对含量之和是水提物中的1.32倍。该方法简便、快捷、灵敏,可为葫芦巴的药效物质基础研究和开发应用提供理论依据。

     

    Abstract: Fenugreek in invigorating kidney, falling blood sugar, etc, have very good medicinal value. Much attention has been paid to saponins, flavonoids, the separation and purification, the compatibility mechanism of these components have been studied in depth, but the preparation is complicated, time-consuming, labor-intensive and poor economic. In this research, compounds in the ethanol extract and the water extract after sugar removal of fenugreek were qualitatively analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOFMS). And the liquiritin was selected as the internal standard for semi-quantitative analysis of all the identified components. Chromatographic separation was performed on a Unitary C18 column (4.6 mm×150 mm×5 μm). Acetonitrile and 0.1% (volume fraction) formic acid solution were used as the mobile phase for gradient elution with the flow rate of 0.5 mL/min. The MS analysis was carried out by the electrospray ionization (ESI) source in negative mode. The qualitative analysis results showed that the total ion chromatograms of the ethanol extract and the water extract after sugar removal of fenugreek were similar with each other, and 36 compounds were identified, including 12 flavonoids and 24 saponin components. Taking typical flavonoids and saponins as examples, the structures of flavonoids and saponin were described in detail, and the possible breaking rules of flavonoids and saponin were deduced. The results of semi-quantitative analysis showed that the relative content of the main components in the two extracts was different. The sum of the relative contents of the flavonoids in the ethanol extract was 1.13 times that in the aqueous extract. The sum of the relative contents of the saponin compounds was 1.32 times that in the aqueous extract. The method is very simple, rapid and sensitive, and the obtained results will provide a theory basis for further research on fenugreek, as well as development and application research of fenugreek.

     

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