title: Fingerprint analysis of Zhimu-Huangbai herb pair and simultaneous determination of its alkaloids, xanthone glycosides and steroidal saponins by HPLC-DAD-ELSD.
abstract: OBJECTIVE
  To develop and validate a high performance liquid chromatography (HPLC) coupled with diode array and evaporative light scattering detectors (DAD-ELSD) method for the quantitative determination and fingerprint analysis of ten active constituents in three chemical classes (namely, xanthone glycosides, steroidal saponins, and alkaloids) in Zhimu-Huangbai herb pair (ZB).
  
  
  METHODS
  Chromatographic separation was performed on a Diamonsil C18 column (4.6 mm × 250 mm, 5 μm, Dikma) by gradient elution using acetic acid in acetonitrile solution at a flow rate of 1.0 mL·min(-1) at 260 nm. The drift tube temperature of ELSD was set to 60 °C and nebulizer gas pressure was 4.0 Bar. Method validation was performed to assure its linearity, limits of detection and quantification, precision, repeatability, stability, and accuracy.
  
  
  RESULTS
  The HPLC-DAD-ELSD method allowed the quantification of ten compounds (phellodendrine, jatrorrhizine, palmatine, berberine, neomangiferin, mangiferin, timosaponin E-I, timosaponin B-II, timosaponin B, and timosaponin A-III), and was successfully applied to fingerprint analysis for ten batches of ZB samples.
  
  
  CONCLUSIONS
  This was the first time to apply the combination of DAD and ELSD for the simultaneous determination of ten active ingredients in ZB. The results showed that the combination of quantitative analysis for marker ingredients and chemical fingerprint for the TCM herb pair provides a potentially powerful, widely introduced, and internationally accepted strategy for assessment of complex TCM formulas.
authors: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
authors: Feng
authors: F
authors: Zhang
authors: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
authors: Qi
authors: Q
authors: Yang
authors: Department of Pharmacognosy, School of Pharmacy, Second Military Medical University, Shanghai 200433, China.
authors: Lian-Na
authors: LN
authors: Sun
authors: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
authors: Shou-Hong
authors: SH
authors: Gao
authors: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
authors: Xia
authors: X
authors: Tao
authors: Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China. Electronic address: chenwanshengsmmu@aliyun.com.
authors: Wan-Sheng
authors: WS
authors: Chen
mesh_terms: D000470:Alkaloids / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000032:analysis*; D006027:Glycosides / Q000032:analysis*; D044004:Xanthones / Q000032:analysis*
chemical_list: D000470:Alkaloids; D004365:Drugs, Chinese Herbal; D006027:Glycosides; D044004:Xanthones; C528424:huangbai; C009689:xanthone
keywords: Alkaloids; Anemarrhena asphodeloides; Chromatographic fingerprint; HPLC-DAD-ELSD; Phellodendron amurense; Simultaneous determination; Steroidal saponins; Xanthone glycosides