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Modified QuEChERS method for phenolic compounds determination in mustard greens (Brassica juncea) using UHPLC-MS/MS
Affiliation:1. Chemistry Department, State University of Maringá (UEM), Maringá, PR, Brazil;2. Institute of Chemistry, State University of Campinas (UNICAMP), Campinas, SP, Brazil
Abstract:A modified QuEChERS method (Quick, Easy, Cheap, Effective, Rugged, and Safe) for the determination of fifteen phenolic compounds in mustard greens (Brassica juncea) using ultra-high-performance liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) analysis was developed. The QuEChERS partitioning step and dispersive solid phase extraction (d-SPE) clean-up sorbents were investigated, aimed at phenolic compound extraction and pigment removal, respectively. QuEChERS acetate version combined with 25 mg of diatomaceous earth (DE) and 5.0 mg of graphitized carbon black (GCB) provided the best conditions for sample preparation of the target compounds. Under the optimized conditions, all phenolic compounds showed good linearity (r ≥ 0.99) over the concentration range of 0.1 to 8000 μg kg−1, and the quantification limits were in the range of 0.06–230 μg kg−1. The spectrophotometric analysis showed that the clean-up step promoted a significant removal of chlorophyll, which is the major pigment present in the sample. Furthermore, antioxidant activity analysis was also carried out after the clean-up step and, together with chromatographic data, showed that no significant retention of the phenolic compounds occurs in the clean-up step. Two mustard greens varieties – Southern Giant Curled (SGC) and Florida Broadleaf (FB) - were analyzed with the proposed method. Seven phenolic compounds (4-hydroxybenzoic, p-coumaric, ferulic and sinapic acids, naringenin, apigenin and kaempferol) were found in both varieties, the greatest abundance being for sinapic acid (1261.5 ± 23 μg kg−1 in SGC and 1235.5 ± 26 μg kg−1 in FB) and ferulic acid (2861 ± 24 μg kg−1 in SGC and 3204.5 ± 45 μg kg−1 in FB).
Keywords:QuEChERS  Antioxidants  Mustard greens  Clean-up  Sorbents  d-SPE  QuEChERS"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0040"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Acronym of Quick, Easy, Cheap, Effective, Rugged, and Safe  UHPLC-MS/MS"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0050"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  ultra-high-performance liquid chromatography coupled to tandem mass spectrometry  d-SPE"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0060"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  dispersive solid phase extraction  DE"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0070"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  diatomaceous earth  GCB"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0080"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  graphitized carbon black  SGC"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0090"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Southern Giant Curled  FB"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0100"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Florida Broadleaf  PSA"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0110"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  primary and secondary amine  LC-MS/MS"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0120"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  liquid chromatography coupled to tandem mass spectrometry  UV–Vis"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0130"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Ultraviolet–visible  DPPH"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0140"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  2,2-diphenyl-1-picryl-hydrazyl-hydrate free radical method  ORAC"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0150"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Oxygen Radical Absorbance Capacity  C18"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0160"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Octadecylsilane  PTFE"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0170"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Polytetrafluoroethylene  AAPH"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0180"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  2,2′-azo-bis(2-amidinopropane) dihydrochloride  SRM"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0190"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  selected reaction monitoring  SD"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0200"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  standard deviation  TE"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0210"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  trolox equivalent  FW"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0220"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  fresh weight  LOD"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0230"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  limit of detection  LOQ"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0240"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  limit of quantification  HPLC-DAD"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0250"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  high performance liquid chromatography with diode array detection  4HB"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0260"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  4-Hydroxybenzoic acid  NAR"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0270"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Naringenin  API"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0280"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Apigenin  KAE"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0290"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Kaempferol  PCU"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0300"  },"  $$"  :[{"  #name"  :"  text"  ,"  $$"  :[{"  #name"  :"  italic"  ,"  _"  :"  p"  },{"  #name"  :"  __text__"  ,"  _"  :"  -Coumaric acid  FER"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0310"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Ferulic acid  SNP"  },{"  #name"  :"  keyword"  ,"  $"  :{"  id"  :"  k0320"  },"  $$"  :[{"  #name"  :"  text"  ,"  _"  :"  Sinapic acid
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