title: Identification of anti-inflammatory constituents from Kalimeris indica with UHPLC-ESI-Q-TOF-MS/MS and GC-MS.
abstract: BACKGROUND
  Kalimeris indica is a Miao׳s medicinal plant in Guizhou province of China employing to treat various inflammation-related diseases in clinical. The study aims to determine the active fractions of K. indica for its anti-inflammatory activity and to identify their chemical constituents.
  
  
  METHODS
  The dried K. indica herb was extracted with 50% aqueous ethanol and then successively separated with macroporous resin and MCI column chromatography to give five fractions (A-E). The anti-inflammatory effects were determined by measuring the NO and TNF-α production in murine macrophage RAW 264.7 cells after exposure to LPS. The chemical constituents of the anti-inflammatory fractions were analyzed by the method of UHPLC-ESI-Q-TOF/MS or GC-MS.
  
  
  RESULTS
  Five fractions (A-E) of different polarities were prepared from the 50% ethanol extract. Factions C and E showed significant inhibition of NO and TNF-α production. Six constituents, namely 3,4-dicaffeoylquinic acid (1), 3,5-dicaffeoylquinic acid (2), 1,5-dicaffeoylquinic acid (3), rutin (4), 1-malonyl-3,5-dicaffeoylquinic acid (5), and 4,5-dicaffeoylquinic acid (6) were identified from the active fraction C by UHPLC-ESI-Q-TOF/MS. Four compounds including 13-tetradecenal (7), (Z,Z)-9,12-octadecadienoic acid (8), (3α)-12-oleanen-3-yl acetate (9), and (+)-3-oxo-urs-12-en-24-oic acid methyl ester (10) were identified from the active fraction E by GC-MS.
  
  
  CONCLUSIONS
  K. indica possessed pronounced anti-inflammatory effect. Dicaffeoylquinic acids and their dirivatives, rutin, as well as oleanolic and fatty acid derivatives are the major constituents and possibly the anti-inflammatory principles of the active fractions of K. indica. All the compounds were identified in K. indica for the first time. The work provided evidence for further development and utilization of K. indica and formed a basis for the establishment of quality control methods and standards for K. indica and its pharmaceutical preparations.
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Rui-Feng
authors.initials: RF
authors.lastname: Zhong
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Guo-Bo
authors.initials: GB
authors.lastname: Xu
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Guizhou Provincial Key Laboratory of Pharmaceutics, Guiyang 550004, Guizhou, China.
authors.forename: Ai-Min
authors.initials: AM
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Huan-Yu
authors.initials: HY
authors.lastname: Guan
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: Guizhou Provincial Key Laboratory of Pharmaceutics, Guiyang 550004, Guizhou, China.
authors.forename: Xun
authors.initials: X
authors.lastname: He
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Jun-Hong
authors.initials: JH
authors.lastname: Liu
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Zhou
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China.
authors.forename: Yong-Jun
authors.initials: YJ
authors.lastname: Li
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Guizhou Provincial Key Laboratory of Pharmaceutics, Guiyang 550004, Guizhou, China.
authors.forename: Yong-Lin
authors.initials: YL
authors.lastname: Wang
authors.affiliation: School of Pharmacy, Guiyang Medical College, Guiyang 550004, Guizhou, China; Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Ministry of Education, Guiyang 550004, Guizhou, China. Electronic address: lshangg@163.com.
authors.forename: Shang-Gao
authors.initials: SG
authors.lastname: Liao
mesh_terms: D000818:Animals; D000893:Anti-Inflammatory Agents / Q000032:analysis*; D019659:Asteraceae / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004365:Drugs, Chinese Herbal / Q000032:analysis / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D009569:Nitric Oxide / Q000032:analysis / Q000037:antagonists & inhibitors; D000067996:RAW 264.7 Cells / Q000737:chemistry / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis / Q000037:antagonists & inhibitors
chemical_list: D000893:Anti-Inflammatory Agents; D004365:Drugs, Chinese Herbal; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide
keywords: (3α)-12-oleanen-3-yl acetate (9), (+)-3-oxo-urs-12-en-24-oic acid methyl ester (10); (Z,Z)-9,12-octadecadienoic acid (PubChem CID: 5280450); 1,5-dicaffeoylquinic acid (PubChem CID 29805); 1-malonyl-3,5-dicaffeoylquinic acid (PubChem CID: 6474367); 3,4-dicaffeoylquinic acid (PubChem CID: 5281780); 3,5-dicaffeoylquinic acid (PubChem CID: 6474310); 4,5-dicaffeoylquinic acid (6), 13-tetradecenal (PubChem CID: 522841); Active fractions; Anti-inflammatory activity; Asteraceae; Dicaffeoylquinic acids; Kalimeris indica; Rutin (PubChem CID: 5280805)