title: Development and validation of a HPLC-UV-ESI-MS method for the simultaneous quantitation of ten bioactive compounds in Dahuang Fuzi Tang.
abstract: OBJECTIVE
  To develop a high performance liquid chromatography (HPLC) coupled with electrospray ionization mass spectrometry (ESI-MS) and ultraviolet (UV) detector method for the acid-alkaline simultaneous determination of ten bioactive compounds, and analyze the effect of compatible medicinal plants on the concentration of components in Dahuang Fuzi Tang (DFT).
  
  
  METHODS
  The chromatographic separation was performed on a Hypersil BDS C18 analytical column by gradient elution with acetonitrile and formate buffer (containing 0.15% formic acid, V/V) at 25 °C with a flow rate of 1.0 mL·min(-1) and UV detection at 280 nm. Four of the ten compounds in DFT were identified and their MS fragments were elucidated by HPLC-ESI-MS, and the contents of the six compounds were determined by HPLC-UV.
  
  
  RESULTS
  All calibration curves showed good linear regression (r(2) ≥ 0.9990). The limits of detection and limits of quantification were 0.021-0.155 -g·mL(-1) and 0.076-0.520 -g·mL(-1), respectively. Overall precision RSD (intra-day and inter-day) were less than 2.96%, and the average recoveries were 98.35%-101.45%, with RSD ranging from 1.54% to 3.01% for the analytes.
  
  
  CONCLUSIONS
  The developed method can be applied for the quality control and provide analytical evidence on the chemical basis and combinational principles of DFT.
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Guo
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Huan
authors.initials: H
authors.lastname: Li
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China; Nanjing Haichang Chinese Medicine Group Corporation, Nanjing 210061, China. Electronic address: liuxiao04_0@163.com.
authors.forename: Xiao
authors.initials: X
authors.lastname: Liu
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Hao
authors.initials: H
authors.lastname: Cai
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wu
authors.affiliation: College of Pharmacy, Nanjing University of Chinese Medicine, Nanjing 210023, China; Engineering Center of State Ministry of Education for Standardization of Chinese Medicine Processing, Nanjing University of Chinese Medicine, Nanjing 210023, China; Nanjing Haichang Chinese Medicine Group Corporation, Nanjing 210061, China. Electronic address: bccai@126.com.
authors.forename: Bao-Chang
authors.initials: BC
authors.lastname: Cai
mesh_terms: D031841:Aconitum / Q000737:chemistry*; D030023:Asarum / Q000737:chemistry*; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000737:chemistry*; D057230:Limit of Detection; D011786:Quality Control; D015203:Reproducibility of Results; D012250:Rheum / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013056:Spectrophotometry, Ultraviolet / Q000379:methods*
chemical_list: D004365:Drugs, Chinese Herbal
keywords: Dahuang Fuzi Tang; HPLC-UV-ESI-MS; Simultaneous analysis