Analytical and Bioanalytical Chemistry

, Volume 410, Issue 15, pp 3607–3619 | Cite as

Characterization of bioactive compounds of Annona cherimola L. leaves using a combined approach based on HPLC-ESI-TOF-MS and NMR

  • Elixabet Díaz-de-Cerio
  • Luis Manuel Aguilera-Saez
  • Ana María Gómez-Caravaca
  • Vito Verardo
  • Alberto Fernández-Gutiérrez
  • Ignacio Fernández
  • David Arráez-Román
Research Paper
Part of the following topical collections:
  1. Discovery of Bioactive Compounds


Annona cherimola Mill. (cherimoya) has widely been used as food crop. The leaves of this tree possess several health benefits, which are, in general, attributed mainly to its bioactive composition. However, literature concerning a comprehensive characterization based on a combined approach, which consists of nuclear magnetic resonance (NMR) and high-performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-TOF-MS), from these leaves is scarce. Thus, the aim of this work was to study the polar profile of full extracts of cherimoya leaves by using these tools. Thus, a total of 77 compounds have been characterized, 12 of which were identified by both techniques. Briefly, 23 compounds were classified as amino acids, organic acids, carbohydrates, cholines, phenolic acid derivatives, and flavonoids by NMR, while 66 metabolites were divided into sugars, amino acids, phenolic acids and derivatives, flavonoids, phenylpropanoids, and other polar compounds by HPLC-TOF-MS. It is worth mentioning that different solvent mixtures were tested and the total phenolic content in the extracts quantified (TPC via HPLC-TOF-MS). The tendency observed was EtOH/water 80/20 (v/v) (17.0 ± 0.2 mg TPC/g leaf dry weight (d.w.)) ≥ acetone/water 70/30 (v/v) (16.1 ± 0.7 mg TPC/g leaf d.w.) > EtOH/water 70/30 (v/v) (14.0 ± 0.3 mg TPC/g leaf d.w.) > acetone/water 80/20 (v/v) (13.5 ± 0.4 mg TPC/g leaf d.w.). Importantly, flavonoids derivatives were between 63 and 76% of the TPC in those extracts. Major compounds were sucrose, glucose (α and β), and proline, and chlorogenic acid and rutin for NMR and HPLC-TOF-MS, respectively.

Graphical abstract

The combined use of LC-HRMS and NMR is a potential synergic combination for a comprehensive metabolite composition of cherimoya leaves


Annona cherimola leaves HPLC-TOF-MS Nuclear magnetic resonance Phenolic compounds Natural compounds 



Vito Verardo thanks the MINECO for his “Ramon y Cajal” contract.


This work was funded by projects AGL2015-67995-C3-2-R (Spanish Ministry of Science and Innovation), P11-CTS-7625, and P12-FQM-2668 (Andalusian Regional Government Council of Innovation and Science).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human or animal subjects.

Informed consent

Informed consent was not applicable.

Supplementary material

216_2018_1051_MOESM1_ESM.pdf (219 kb)
ESM 1 (PDF 218 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Elixabet Díaz-de-Cerio
    • 1
  • Luis Manuel Aguilera-Saez
    • 2
  • Ana María Gómez-Caravaca
    • 1
  • Vito Verardo
    • 3
    • 4
  • Alberto Fernández-Gutiérrez
    • 1
  • Ignacio Fernández
    • 2
  • David Arráez-Román
    • 1
    • 5
  1. 1.Department of Analytical Chemistry, Faculty of SciencesUniversity of GranadaGranadaSpain
  2. 2.Department of Chemistry and Physics, Research Centre for Agricultural and Food Biotechnology (CIAIMBITAL)University of AlmeriaAlmeriaSpain
  3. 3.Department of Nutrition and Food ScienceUniversity of Granada, Campus Universitario de CartujaGranadaSpain
  4. 4.Institute of Nutrition and Food Technology ‘José Mataix’, Biomedical Research CentreUniversity of GranadaGranadaSpain
  5. 5.Health Science Technological ParkResearch and Development Functional Food CentreGranadaSpain

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