Screening and identification of potential bioactive components in a combined prescription of Danggui Buxue decoction using cell extraction coupled with high performance liquid chromatography.

A cell extraction coupled with high-performance liquid chromatography (HPLC) analysis has been developed to screen the potential bioactive components in combined prescription of Danggui Buxue decoction. The method was validated by using HL-7702 cells, RAW 264.7 cells and Caco-2 cell extraction. According to the hypothesis that, when cells are incubated together with the extract of traditional Chinese medicines (TCMs), the potential bioactive compounds in the extract should selectively combine with the cells, cell-combining compounds were analyzed by HPLC-diode array-evaporative light scattering detectors. Their structures were elucidated in comparison with available reference compounds, and further confirmed by HPLC-electrospray ionization time-of-flight mass spectrometry with accurate mass measurement. The results demonstrated that nine compounds were combined with the HL-7702 cells, seven with RAW 264.7 cells, and thirteen with the Caco-2 cell line. In view of the two key steps of drugs action, absorption by intestinal epithelium cells and interaction with target cells, this rapid and reliable method could be utilized to predict the bioactive constituents in TCMs, and it was in agreement with the characteristics of combined prescriptions of TCMs as multi-components, multi-target sites and multi-channel actions.

[1]  T. Furuya,et al.  Two triglycosidic triterpene astragalosides from hairy root cultures of Astragalus membranaceus , 1995 .

[2]  P. Artursson,et al.  Correlation between oral drug absorption in humans and apparent drug permeability coefficients in human intestinal epithelial (Caco-2) cells. , 1991, Biochemical and biophysical research communications.

[3]  Ping Li,et al.  Separation of compounds interacting with liposome membrane in combined prescription of traditional Chinese medicines with immobilized liposome chromatography. , 2005, Journal of pharmaceutical and biomedical analysis.

[4]  T. Furuya,et al.  Astragalosides from hairy root cultures of Astragalus membranaceus , 1994 .

[5]  Ping Li,et al.  Live cell extraction and HPLC-MS analysis for predicting bioactive components of traditional Chinese medicines. , 2006, Journal of pharmaceutical and biomedical analysis.

[6]  T. Dong,et al.  Chemical analysis of Radix Astragali (Huangqi) in China: a comparison with its adulterants and seasonal variations. , 2002, Journal of agricultural and food chemistry.

[7]  Xin Li,et al.  Screening and analysis of permeable compounds in Radix Angelica Sinensis with immobilized liposome chromatography. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[8]  Ma Xiao-feng,et al.  Flavonoid constituents of Astragalus membranaceus var. mongholicus , 2005 .

[9]  Ling Yi,et al.  Analysis of interaction property of bioactive components in Danggui Buxue Decoction with protein by microdialysis coupled with HPLC-DAD-MS. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[10]  P. Artursson Cell cultures as models for drug absorption across the intestinal mucosa. , 1991, Critical reviews in therapeutic drug carrier systems.

[11]  G. Lin,et al.  Simultaneous analysis of seventeen chemical ingredients of Ligusticum chuanxiong by on-line high performance liquid chromatography-diode array detector-mass spectrometry. , 2003, Planta medica.

[12]  Y. Oshima,et al.  ISOFLAVANS AND A PTEROCARPAN FROM ASTRAGALUS MONGHOLICUS , 1991 .

[13]  Xian-guo He,et al.  Liquid chromatographic–electrospray mass spectrometric study of the phthalides of Angelica sinensis and chemical changes of Z-ligustilide , 1998 .

[14]  Ping Li,et al.  Screening and identification of permeable components in a combined prescription of Danggui Buxue decoction using a liposome equilibrium dialysis system followed by HPLC and LC-MS. , 2006, Journal of separation science.

[15]  Zhaohui Wu,et al.  Knowledge discovery in traditional Chinese medicine: State of the art and perspectives , 2006, Artif. Intell. Medicine.

[16]  J. Yu,et al.  Separation and identification of the phthalic anhydride derivatives of Liqusticum Chuanxiong Hort by GC-MS, TLC, HPLC-DAD, and HPLC-MS. , 2002, Journal of chromatographic science.

[17]  C. Pidgeon,et al.  IAM chromatography: an in vitro screen for predicting drug membrane permeability. , 1995, Journal of medicinal chemistry.

[18]  T. Furuya,et al.  Cycloartane triterpene glycosides from the hairy root cultures of Astragalus membranaceus. , 1994, Phytochemistry.

[19]  G. Cordell,et al.  Liquid chromatography-electrospray ionization mass spectrometry study of the flavonoids of the roots of Astragalus mongholicus and A. membranaceus. , 2000, Journal of chromatography. A.