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报道了用高表达表皮生长因子受体细胞膜色谱与高效液相色谱/质谱在线联用方法(EGFR/CMC-online-HPLC/MS)快速筛选发现中药独活中的活性成分.实验中,采用高表达EGFR的细胞膜制备色谱固定相,建立EGFR/细胞膜色谱(EGFR/CMC)模型,利用柱切换和固相萃取技术,将EGFR/CMC模型与高效液相色谱/质谱(HPLC/MS)在线联用,构成一种新的可同时“识别-鉴定”目标成分的二维色谱系统,并应用于快速筛选独活中具有抗EGFR活性的目标成分.结果发现独活中的蛇床子素具有与对照药物达沙替尼类似的色谱保留特性,能够作用于EGFR;同时MTT及Elisa分析实验证实蛇床子素对HEK293EGFR细胞增殖及EGFR表达均有抑制作用.本文建立的EGFR/CMC-online-HPLC/MS二维色谱方法,可以选择性地从中药复杂体系中快速“识别-鉴定”目标组分,且筛选结果与特定生物效应显著相关.
Reported the rapid screening of EGFR / CMC-online-HPLC / MS using high expression of epidermal growth factor receptor cell membrane chromatography combined with high performance liquid chromatography / mass spectrometry (online-HPLC / MS) (EGFR / CMC) model was established and the EGFR / CMC model was combined with high performance liquid chromatography / mass spectrometry (HPLC / MS) using column switching and solid phase extraction (SPE / MS) A new two-dimensional chromatographic system which can simultaneously identify the target components of “identification - identification ” and applied to the rapid screening of the target components of anti-EGFR activity alone .Results found that osthole alone with the control drug up to The similar chromatographic retention properties of cediranib could act on EGFR, and both MTT and Elisa analysis confirmed that osthole had inhibitory effect on the proliferation and EGFR expression of HEK293EGFR cells.In the present study, EGFR / CMC-online-HPLC / MS two-dimensional Chromatographic methods can be used to rapidly “identify-identify” target components from complex systems of traditional Chinese medicine, and the screening results are significantly associated with specific biological effects.