Neutral Loss Scan - Based Strategy for Integrated Identification of Amorfrutin Derivatives, New Peroxisome Proliferator-Activated Receptor Gamma Agonists, from Amorpha Fruticosa by UPLC-QqQ-MS/MS and UPLC-Q-TOF-MS

  • Chu Chen
  • Ying Xue
  • Qing-Miao Li
  • Yan Wu
  • Jian Liang
  • Lin-Sen Qing
Research Article


Amorfrutins with a 2-hydroxybenzoic acid core structure are promising natural PPARγ agonists with potent antidiabetic activity. Owing to the complex matrix and low concentration in botanical material, the identification of unknown amorfrutins remains a challenge. In the present study, a combined application of UPLC-Q-TOF-MS and UPLC-QqQ-MS was developed to discover unknown amorfrutins from fruits of Amorpha fruticosa. First, reference compounds of amorfrutin A (AA), amorfrutin B (AB), and 2-carboxy-3,5-dihydroxy-4-geranylbibenzyl (AC) were analyzed using UPLC-Q-TOF-MS to reveal the characteristic fragment ions and the possible neutral loss. Second, the extract of A. fruticosa was separated and screened by UPLC-QqQ-MS using neutral loss scan to find out suspect compounds associated with the specified neutral fragment Δm/z 44. Third, the extract was re-analyzed by UPLC-Q-TOF-MS to obtain the exact mass of quasi-molecular ion and fragment ions of each suspect compound, and to subsequently calculate their corresponding molecular formulas. Finally, according to the molecular formula of suspect compound and its fragment ions and comparing with literature data, structure elucidation of four unidentified amorfrutins was achieved. The results indicated that the combination of QqQ-MS neutral loss scan and Q-TOF-MS molecular formula calculation was proven to be a powerful tool for unknown natural product identification, and this strategy provides an effective solution to discover natural products or metabolites of trace content.

Graphical Abstract


Amorfrutins PPARγ agonists Amorpha fruticosa UPLC-QqQ-MS/MS UPLC-Q-TOF-MS Neutral loss scan 



This work was financially supported by the National Natural Science Foundation of China (21202161) and the Basic Scientific Research Foundation of Sichuan Academy of Chinese Medicine Sciences (A-2016N-38).

Supplementary material

13361_2018_1891_MOESM1_ESM.pdf (377 kb)
ESM 1 (PDF 376 kb)


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Copyright information

© American Society for Mass Spectrometry 2018

Authors and Affiliations

  1. 1.CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization and Ecological Restoration and Biodiversity Conservation Key Laboratory of Sichuan Province, Chengdu Institute of BiologyChinese Academy of SciencesChengduChina
  2. 2.Sichuan Academy of Chinese Medicine SciencesChengduChina
  3. 3.Sichuan Provincial Center for Disease Control and PreventionChengduChina
  4. 4.Natural Products Research Center, Chengdu Institute of BiologyChinese Academy of SciencesChengduChina

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