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目的:建立HPLC-DAD-ESI-MS分析鉴定穿心莲二萜内酯有效部位的主要化学成分的方法,并对其药效学进行研究。方法:Agilent SB-C18色谱柱(2.1 mm×150 mm,5μm);流动相A为乙腈,流动相B为水,60 min梯度洗脱;流速0.4 m L·min-1;进样5μL;检测波长205 nm。柱温为25℃;采用正离子模式,m/z扫描范围50~1 000;利用2种肿瘤细胞株造活体动物模型,对有效部位进行抗肿瘤方面的初步药效学研究。结果:穿心莲二萜内酯有效部位中有5种二萜内酯类化学成分获得了良好的分离与鉴定。药效学实验表明,穿心莲二萜内酯有效部位对肿瘤有一定的抑制作用。结论:建立了一种基于HPLC-DAD-ESI-MS技术对穿心莲二萜内酯有效部位的化学成分进行快速鉴别的方法,并进行了初步药效学研究,为穿心莲二萜内酯有效部位的质量控制及体内的深入研究奠定了基础。
OBJECTIVE: To establish a HPLC-DAD-ESI-MS method for the determination of the main chemical constituents in the effective fractions of diterpene lactone, and to study its pharmacodynamics. Method: Agilent SB-C18 column (2.1 mm × 150 mm, 5 μm); mobile phase A was acetonitrile, mobile phase B was water and eluted with 60 min gradient; flow rate was 0.4 m L · min-1; Wavelength 205 nm. The column temperature was 25 ℃. In the positive mode, the m / z scan range was 50-1 000. The antitumor efficacy of the two tumor cell lines was evaluated in animal models. Results: Five diterpene lactones from the effective fractions of andrographolide were well separated and identified. Pharmacodynamic experiments show that the andrographolide diterpene lactone effective part of the tumor has a certain inhibitory effect. Conclusion: A HPLC-DAD-ESI-MS technique was developed for rapid identification of the active ingredients of andrographolide diterpene lactone. A preliminary pharmacodynamic study was conducted to determine the effective components of the diterpene lactone Quality control and in-depth study laid the foundation.