title: Combined Application of UHPLC-QTOF/MS, HPLC-ELSD and1H-NMR Spectroscopy for Quality Assessment of DA-9801, A Standardised Dioscorea Extract.
abstract: BACKGROUND
  DA-9801, a standardised 50% aqueous ethanolic extract of a mixture of Dioscorea japonica and D. nipponica, is a botanical drug candidate for the treatment of diabetic neuropathy, which finished its US phase II clinical trials recently. An advanced quality control method is needed for further development of DA-9801, considering its high contents of both primary and secondary metabolites.
  
  
  OBJECTIVE
  Development of a quality assessment strategy for DA-9801, based on the combination of UHPLC-QTOF/MS, HPLC-ELSD, and1H-NMR spectroscopy.
  
  
  METHODS
  The method was developed and tested with 15 batch products of DA-9801. The steroidal saponins of DA-9801 were tentatively identified by UHPLC-QTOF/MS and were quantified with the validated HPLC-ELSD method. Primary metabolites of DA-9801 were identified and profiled using1H-NMR spectrometry. The batch-to-batch equivalence of DA-9801 was tested with the1H-NMR spectra using spectral binning, correlation analysis, and principal component analysis.
  
  
  RESULTS
  Six major saponins of DA-9801 were tentatively identified by UHPLC-QTOF/MS. Among them, protodioscin and dioscin were quantified by the validated HPLC-ELSD method. Twenty-six metabolites were identified in1H-NMR spectra. The similarity between DA-9801 batches could be evaluated with the NMR spectra of DA-9801. The1H-NMR method also revealed that two Dioscorea species contributed distinct amino acids to the contents of DA-9801.
  
  
  CONCLUSIONS
  This study validates the effectiveness of UHPLC-QTOF/MS, HPLC-ELSD, and1H NMR-combined method for quality control of DA-9801 and its crude materials. Copyright © 2016 John Wiley & Sons, Ltd.
authors.affiliation: College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University, Seoul, 08826, Republic of Korea.
authors.forename: Kyo Bin
authors.initials: KB
authors.lastname: Kang
authors.affiliation: Research Planning & Management Department, Dong-A ST Co., Ltd., Yongin, 17073, Republic of Korea.
authors.forename: Jayoung
authors.initials: J
authors.lastname: Ryu
authors.affiliation: Research Planning & Management Department, Dong-A ST Co., Ltd., Yongin, 17073, Republic of Korea.
authors.forename: Youngwoong
authors.initials: Y
authors.lastname: Cho
authors.affiliation: Research Planning & Management Department, Dong-A ST Co., Ltd., Yongin, 17073, Republic of Korea.
authors.forename: Sang-Zin
authors.initials: SZ
authors.lastname: Choi
authors.affiliation: Research Planning & Management Department, Dong-A ST Co., Ltd., Yongin, 17073, Republic of Korea.
authors.forename: Miwon
authors.initials: M
authors.lastname: Son
authors.affiliation: College of Pharmacy and Research Institute of Pharmaceutical Science, Seoul National University, Seoul, 08826, Republic of Korea.
authors.forename: Sang Hyun
authors.initials: SH
authors.lastname: Sung
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D027742:Dioscorea / Q000737:chemistry; D004144:Diosgenin / Q000031:analogs & derivatives / Q000032:analysis; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D028321:Plant Preparations / Q000032:analysis* / Q000592:standards; D011786:Quality Control; D015203:Reproducibility of Results; D012503:Saponins / Q000032:analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation / Q000379:methods
chemical_list: C562027:DA-9801; D028321:Plant Preparations; D012503:Saponins; C019357:dioscin; C459619:protodioscin; D004144:Diosgenin
keywords: 1H-NMR spectroscopy; DA-9801; HPLC-ELSD; UHPLC-QTOF/MS; quality assessment