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Objective: Compound traditional Chinese medicine (CTCM) with the application of compatibility from multiple active ingredients with multiple-specific targets can achieve a synergistic effect on cancer therapy. This study is aimed to observe the compatibility effects of Aidi injection on the treatment of hepatocellular carcinoma and to explore the mechanism of CTCM. Methods: Aidi injection is a clinical compound prescription containing Mylabris, Ginseng, Astragalus, and Acanthopanax, which can inhibit tumor growth and induce apoptosis. In this study, the anticancer activity of Aidi injection, as well as its disassembled and combined compositions, had been evaluated by varying levels of polyamine biomarkers on human hepatoma Hep-G2 cells detected using ultrahigh-performance liquid chromatography-tandem mass spectrometry. Results: According to the different variations in polyamine levels, it was revealed that Mylabris and Ginseng had an antitumor effect, while Astragalus acted as an assistant and Acanthopanax had weak anticancer activity. The increased level of polyamines in Hep-G2 cells had been found in HL-7702 cells. On combining Mylabris and Ginseng, polyamine levels went close to the normal level, which was even more marked when Astragalus was added. Aidi injection acted like the combination of Mylabris, Ginseng, and Astragalus. Conclusions: This study established a quantitative evaluation of the compatibility effects of Aidi injection based on polyamine biomarkers and evaluated the consistency of its anticancer effect, providing a manner to research the efficacy evaluation of CTCM. Moreover, the correlation between polyamine metabolism and anticancer activity can be used in anticancer drug screening.

A novel, improved, and comprehensive method for quality evaluation and discrimination of Herba Leonuri has been developed and validated based on normal- and reversed-phase chromatographic methods. To identify Herba Leonuri, normal- and reversed-phase high-performance thin-layer chromatography fingerprints were obtained by comparing the colors and Rf values of the bands, and reversed-phase high-performance liquid chromatography fingerprints were obtained by using an Agilent Poroshell 120 SB-C18 within 28 min. By similarity analysis and hierarchical clustering analysis, we show that there are similar chromatographic patterns in Herba Leonuri samples, but significant differences in counterfeits and variants. To quantify the bio-active components of Herba Leonuri, reversed-phase high-performance liquid chromatography was performed to analyze syringate, leonurine, quercetin-3-O-robiniaglycoside, hyperoside, rutin, isoquercitrin, wogonin, and genkwanin simultaneously by single standard to determine multi-components method with rutin as internal standard. Meanwhile, normal-phase high-performance liquid chromatography was performed by using an Agilent ZORBAX HILIC Plus within 6 min to determine trigonelline and stachydrine using trigonelline as internal standard. Innovatively, among these compounds, bio-active components of quercetin-3-O-robiniaglycoside and trigonelline were first determined in Herba Leonuri. In general, the method integrating multi-chromatographic analyses offered an efficient way for the standardization and identification of Herba Leonuri.