title: Developing an activity and absorption-based quality control platform for Chinese traditional medicine: Application to Zeng-Sheng-Ping(Antitumor B).
abstract: BACKGROUND
  Zeng-Sheng-Ping (ZSP), also called antitumor B, is a marketed Chinese traditional medicine used for cancer prevention.
  
  
  OBJECTIVE
  Currently, for the quality control of Chinese traditional medicines, marker compounds are not selected based on bioactivities and pharmaceutical behaviors in most of the cases. Therefore, even if the "quality" of the medicine is controlled, the pharmacological effect could still be inconsistent. The aim of this study is to establish an activity and absorption-based platform to select marker compound(s) for the quality control of Chinese traditional medicines.
  
  
  METHODS
  We used ZSP as a reference Chinese traditional medicine to establish the platform. Activity guided fractionation approach was used to purify the major components from ZSP. NMR and MS spectra were used to elucidate the structure of the isolated compounds. MTT assay against oral carcinoma cell line (SCC2095) was performed to evaluate the activities. UPLC-MS/MS was used to quantify the pure compounds in ZSP and the active fraction. The permeabilities of the identified compounds were evaluated in the Caco-2 cell culture model. The intracellular accumulation of the isolated compounds was evaluated in the SCC2095 cells.
  
  
  RESULTS
  The major compounds were identified from ZSP. The contents, anti-proliferation activities, permeabilities, and intracellular accumulations of these compounds were also evaluated. The structure of these purified compounds were identified by comparing the NMR and MS data with those of references as rutaevine (1), limonin (2), evodol (3), obacunone (4), fraxinellone (5), dictamnine (6), maackiain (7), trifolirhizin (8), and matrine (9). The IC50 of compounds 5, 6, and 7 against SCC2095 cells were significantly lower than that of ZSP. The uptake permeability of compounds 5, 6, and 7 were 2.58 ± 0.3 × 10(-5), 4.33 ± 0.5 × 10(-5), and 4.27 ± 0.8 × 10(-5) respectively in the Caco-2 cell culture model. The intracellular concentrations of these compounds showed that compounds 5, 6, and 7 were significantly accumulated inside the cells.
  
  
  CONCLUSIONS
  Based on the activity against oral carcinoma cell line as well as the absorption permeability, compound 5, 6, and 7 are selected as quality control markers for ZSP. An activity and absorption-based platform was established and successfully used for the quality control of ZSP.
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 1441 Moursund Street, Houston, TX 77030, USA.
authors.forename: Taijun
authors.initials: T
authors.lastname: Yin
authors.affiliation: Hubei University of University affiliated Taihe Hospital, 30 South Renmin Road, Shiyan, Hubei, China.
authors.forename: Guanyi
authors.initials: G
authors.lastname: Yang
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 1441 Moursund Street, Houston, TX 77030, USA.
authors.forename: Yong
authors.initials: Y
authors.lastname: Ma
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 1441 Moursund Street, Houston, TX 77030, USA.
authors.forename: Beibei
authors.initials: B
authors.lastname: Xu
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 1441 Moursund Street, Houston, TX 77030, USA; Hubei University of University affiliated Taihe Hospital, 30 South Renmin Road, Shiyan, Hubei, China.
authors.forename: Ming
authors.initials: M
authors.lastname: Hu
authors.affiliation: Medical College of Wisconsin Cancer Center and Department of Pharmacology and Toxicology, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.
authors.forename: Ming
authors.initials: M
authors.lastname: You
authors.affiliation: Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, The University of Houston, 1441 Moursund Street, Houston, TX 77030, USA; Hubei University of University affiliated Taihe Hospital, 30 South Renmin Road, Shiyan, Hubei, China. Electronic address: sgao3@uh.edu.
authors.forename: Song
authors.initials: S
authors.lastname: Gao
mesh_terms: D000470:Alkaloids / Q000032:analysis / Q000302:isolation & purification; D001572:Benzofurans / Q000032:analysis / Q000302:isolation & purification; D001584:Benzoxepins / Q000032:analysis / Q000302:isolation & purification; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D004365:Drugs, Chinese Herbal / Q000737:chemistry* / Q000494:pharmacology*; D005960:Glucosides / Q000032:analysis / Q000302:isolation & purification / Q000493:pharmacokinetics; D006576:Heterocyclic Compounds, 4 or More Rings / Q000032:analysis / Q000302:isolation & purification / Q000493:pharmacokinetics; D006801:Humans; D036701:Limonins / Q000032:analysis / Q000302:isolation & purification; D008516:Medicine, Chinese Traditional / Q000592:standards*; D010539:Permeability; D036343:Pterocarpans / Q000032:analysis / Q000302:isolation & purification / Q000493:pharmacokinetics; D011786:Quality Control; D011804:Quinolines / Q000032:analysis / Q000302:isolation & purification / Q000493:pharmacokinetics; D011807:Quinolizines / Q000032:analysis / Q000302:isolation & purification; D014315:Triterpenes / Q000032:analysis / Q000302:isolation & purification; D000093842:Matrines; D000094162:Phytoalexins
chemical_list: D000470:Alkaloids; D001572:Benzofurans; D001584:Benzoxepins; D004365:Drugs, Chinese Herbal; D005960:Glucosides; D006576:Heterocyclic Compounds, 4 or More Rings; D036701:Limonins; D036343:Pterocarpans; D011804:Quinolines; D011807:Quinolizines; D014315:Triterpenes; D000093842:Matrines; C027128:Antitumor B; C526230:trifolirhizin; C075391:fraxinellone; C079993:rutaevin; C067207:obacunone; C026398:dictamnine; C001546:limonin; C001449:inermin; D000094162:Phytoalexins
keywords: Absorption; Anti-proliferation; Oral cancer; Quality control; Zeng-Sheng-Ping