Mei Zhanghao, Li Huifen, Li Tingting, Taoli Huiping, Pan Linmei
Department of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Jiangsu Research Center of Botanical Medicine Refinement Engineering, Nanjing 210023, China.
Molecules. 2025 Mar 26;30(7):1478. doi: 10.3390/molecules30071478.
Ledeb. (APL), a traditional Chinese herb frequently employed by Professor Zhou Zhongying, a master of traditional Chinese medicine, for colorectal cancer treatment, is rich in polyphenols with potential anti-tumor properties. To elucidate its bioactive components, this study developed a two-step purification process combining macroporous resin adsorption and liquid-liquid extraction to enrich polyphenols from APL (APLs). The adsorption/desorption mechanisms of APLs on macroporous resins were systematically investigated through resin screening, adsorption kinetics, and thermodynamics. The Langmuir isotherm model confirmed the adsorption process as spontaneous and exothermic. Pseudo-second-order kinetics effectively described the adsorption behavior of D101 resin. Optimized adsorption and column elution parameters were established, followed by liquid-liquid extraction for further purification. The components were compared and analyzed by ultra-performance liquid chromatography and quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-Zeno-TOF-MS/MS). It was preliminarily identified that 29 polyphenols were mainly concentrated in water-saturated n-butanol (BEA) and ethyl acetate (ECA) extract fractions. Quantitative analysis using ultra-high performance liquid chromatography-triple quadrupole liquid chromatography-mass spectrometry (UHPLC-C-QTRAP-MS/MS) revealed higher contents of catechin (66.67 ± 1.33 ng·mg), hyperoside (382.56 ± 3.65 ng·mg), and chlorogenic acid (10.60 ± 0.05 ng·mg) in BEA compared to ECA (46.00 ± 2.00, 239.40 ± 2.60, and 3.42 ± 0.01 ng·mg, respectively). In vitro experiments demonstrated that BEA exhibited superior antiproliferative activity (IC: 434.5 μg·mL) and significantly inhibited CT26 tumor cell migration compared to ECA (IC: 672.5 μg·mL). The enhanced biological activity of BEA may be due to its higher polyphenol content, suggesting that these compounds mediate their anti-tumor effects through different biochemical pathways. This work lays the foundation for exploring the multi-target mechanism of anti-tumor effects of APLs.
中药白头翁（APL）是中医大师周仲瑛教授常用于治疗结直肠癌的传统草药，富含具有潜在抗肿瘤特性的多酚类物质。为阐明其生物活性成分，本研究开发了一种两步纯化工艺，结合大孔树脂吸附和液液萃取，从白头翁中富集多酚类物质（APLs）。通过树脂筛选、吸附动力学和热力学系统研究了APLs在大孔树脂上的吸附/解吸机制。Langmuir等温线模型证实吸附过程是自发的且放热的。准二级动力学有效地描述了D101树脂的吸附行为。建立了优化的吸附和柱洗脱参数，随后进行液液萃取进一步纯化。通过超高效液相色谱和四极杆飞行时间串联质谱（UPLC-Q-Zeno-TOF-MS/MS）对成分进行比较和分析。初步鉴定出29种多酚类物质主要集中在水饱和正丁醇（BEA）和乙酸乙酯（ECA）萃取馏分中。使用超高效液相色谱-三重四极杆液相色谱-质谱联用仪（UHPLC-C-QTRAP-MS/MS）进行定量分析显示，与ECA（分别为46.00±2.00、239.40±2.60和3.42±0.01 ng·mg）相比，BEA中儿茶素（66.67±1.33 ng·mg）、金丝桃苷（382.56±3.65 ng·mg）和绿原酸（10.60±0.05 ng·mg）的含量更高。体外实验表明，与ECA（IC：672.5 μg·mL）相比，BEA表现出更强的抗增殖活性（IC：434.5 μg·mL），并显著抑制CT26肿瘤细胞迁移。BEA增强的生物活性可能归因于其较高的多酚含量，表明这些化合物通过不同的生化途径介导其抗肿瘤作用。这项工作为探索APLs抗肿瘤作用的多靶点机制奠定了基础。
