title: Chromatographic isolation of nanoparticles from Ma-Xing-Shi-Gan-Tang decoction and their characterization.
abstract: BACKGROUND
  The herbal decoction is a complex dispersion system containing solutes, colloid, aggregates, emulsions and precipitates. In which phase bioactive phytochemicals are dispersed determines their delivery, action and metabolism. This study took ephedrine, a well-studied and widely used phytochemical, as an example to elucidate its exact distribution in the phases of Ma-Xing-Shi-Gan-Tang decoction (MXSGT), which is an Ephedra sinica Stapf. containing traditional Chinese medicinal formula, and the biological meaning of this distribution correspondingly. It may provide an important update to the safety and efficacy assessment of the herbal decoction and its active phytochemicals.
  
  
  METHODS
  In this study, the decoction was fractionated with size-exclusion chromatography coupled with multi-angle laser light scattering detector. The morphology of fractionated nanoparticles was observed with AFM and SEM. The bioactivities of the decoction, the ephedrine alkaloids loaded NPs (prepared by chromatography isolation) and the synthetic ephedrine were assessed by cell proliferation tests using five cell lines, namely Caco-2, L-02, Hep-G2, NR-8383, and Hela-229.
  
  
  RESULTS
  Nanoparticles with radii of gyration ranged from 50 to 150 nm were isolated, in spherical shape. Further analysis of nanoparticles on the subsequent reversed phase chromatography revealed that the majority of ephedrine (99.7%) and pseudoephedrine (95.5%) were associated with these nanoparticles, rather than dispersed freely in the real solution. The addition of both the herbal decoction and the separated ephedrine-loaded nanoparticles reserved higher cell viability/proliferation than that of the sole synthetic ephedrine among the Caco-2, L-02, Hep-G2, and NR-8383 cells. In contrast, the nanoparticles reduced the proliferating power of ephedrine on Hela-229 cells. In general, the ephedrine-loaded NPs conducted the intermediate influences on the cell viability, in either way.
  
  
  CONCLUSIONS
  The colloidal nanoparticles were separated from the decoction. The association of ephedrine alkaloids with nanoparticles was demonstrated and may have changed the bioactivity of the alkaloids. The naturally occurred colloidal nanoparticles may play an important role in the pharmacological properties of both the decoction and its active phytochemicals, therefore warrant further studies.
authors.affiliation: CAS.SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310035, China; Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China.
authors.forename: Jianwu
authors.initials: J
authors.lastname: Zhou
authors.affiliation: CAS.SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310035, China; Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China.
authors.forename: Guanzhen
authors.initials: G
authors.lastname: Gao
authors.affiliation: Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China.
authors.forename: Qiuping
authors.initials: Q
authors.lastname: Chu
authors.affiliation: CAS.SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310035, China; Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China.
authors.forename: Huiqin
authors.initials: H
authors.lastname: Wang
authors.affiliation: CAS.SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310035, China; Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China.
authors.forename: Pingfan
authors.initials: P
authors.lastname: Rao
authors.affiliation: CAS.SIBS-Zhejiang Gongshang University Joint Centre for Food and Nutrition Sciences, Zhejiang Gongshang University, Hangzhou 310035, China; Institute of Biotechnology, Fuzhou University, Fuzhou 350002, China. Electronic address: lijingke@mail.zjgsu.edu.cn.
authors.forename: Lijing
authors.initials: L
authors.lastname: Ke
mesh_terms: D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D004809:Ephedrine / Q000032:analysis / Q000494:pharmacology; D006801:Humans; D018625:Microscopy, Atomic Force; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles / Q000032:analysis* / Q000648:ultrastructure; D054199:Pseudoephedrine / Q000032:analysis; D051381:Rats
chemical_list: D004365:Drugs, Chinese Herbal; C000591856:ma-xing-shi-gan-tang; D054199:Pseudoephedrine; D004809:Ephedrine
keywords: Cell proliferation; Ephedrine; Ephedrine (PubChem CID: 9294); Ma-Xing-Shi-Gan-Tang decoction; Nanoparticles; Pseudoephedrine; Pseudoephedrine (PubChem CID: 62946); Size-exclusion gel chromatography