, 15:14 | Cite as

Phenolic variation among Chamaecrista nictitans subspecies and varieties revealed through UPLC-ESI(-)-MS/MS chemical fingerprinting

  • Luis Quirós-Guerrero
  • Federico Albertazzi
  • Emanuel Araya-Valverde
  • Rosaura M. Romero
  • Heidy Villalobos
  • Luis Poveda
  • Max Chavarría
  • Giselle Tamayo-CastilloEmail author
Original Article



Comparative analysis of metabolic features of plants has a high potential for determination of quality control of active ingredients, ecological or chemotaxonomic purposes. Specifically, the development of efficient and rapid analytical tools that allow the differentiation among species, subspecies and varieties of plants is a relevant issue. Here we describe a multivariate model based on LC–MS/MS fingerprinting capable of discriminating between subspecies and varieties of the medicinal plant Chamaecrista nictitans, a rare distributed species in Costa Rica.


Determination of the chemical fingerprint was carried out on a LC–MS (ESI-QTOF) in negative ionization mode, main detected and putatively identified compounds included proanthocyanidin oligomers, several flavonoid C- and O-glycosides, and flavonoid acetates. Principal component analysis (PCA), partial least square-discriminant analysis (PLS-DA) and cluster analysis of chemical profiles were performed.


Our method showed a clear discrimination between the subspecies and varieties of Chamaecrista nictitans, separating the samples into four fair differentiated groups: M1 = C. nictitans ssp. patellaria; M2 = C. nictitans ssp. disadena; M3 = C. nictitans ssp. nictitans var. jaliscensis and M4 = C. nictitans ssp. disadena var. pilosa. LC–MS/MS fingerprint data was validated using both morphological characters and DNA barcoding with ITS2 region. The comparison of the morphological characters against the chemical profiles and DNA barcoding shows a 63% coincidence, evidencing the morphological similarity in C. nictitans. On the other hand, genetic data and chemical profiles grouped all samples in a similar pattern, validating the functionality of our metabolomic approach.


The metabolomic method described in this study allows a reliably differentiation between subspecies and varieties of C. nictitans using a straightforward protocol that lacks extensive purification steps.


Chamaecrista nictitans Chemotaxonomy Metabolite fingerprinting LC–MS/MS Chemical fingerprinting 



The authors wish to thank Vicerrectoría de Investigación of the University of Costa Rica, for their financial support (project number 809-B3-082) and the Herbarium of the University of Costa Rica (USJ) for supplying samples from the collection.

Author contributions

LQG, EAV, MC, RMR and GTC wrote the paper; LQG designed and did the LC–MS/MS experiments and MS data analysis; LQG, FA, LP and GTC collected the wild plant material; LP did the botanical identification; RMR did the in vitro culturing of wild plant material and provided greenhouse plantlets for the experiments, FA, HV, EAV and MC performed DNA extraction and ITS2 amplification and sequence analysis, LQG did the data mining and statistical analysis of MS data. GTC, FA and MC did the experimental design of the research.

Compliance with ethical standards

Conflict of interest

There is no conflict of interest.

Ethical statements

This article does not contain any studies with human and/or animal participants performed by any of the authors.

Supplementary material

11306_2019_1475_MOESM1_ESM.doc (2.8 mb)
Supplementary material 1 (DOC 2870 KB)


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Luis Quirós-Guerrero
    • 1
    • 2
  • Federico Albertazzi
    • 3
    • 4
  • Emanuel Araya-Valverde
    • 5
    • 7
  • Rosaura M. Romero
    • 1
    • 2
  • Heidy Villalobos
    • 2
    • 3
  • Luis Poveda
    • 6
  • Max Chavarría
    • 1
    • 2
    • 5
  • Giselle Tamayo-Castillo
    • 1
    • 2
    Email author
  1. 1.Centro de Investigaciones en Productos Naturales (CIPRONA)Universidad de Costa RicaSan JoseCosta Rica
  2. 2.Escuela de QuímicaUniversidad de Costa RicaSan JoseCosta Rica
  3. 3.Centro de Investigación en Biología Celular y Molecular (CIBCM)Universidad de Costa RicaSan JoseCosta Rica
  4. 4.Escuela de BiologíaUniversidad de Costa RicaSan JoseCosta Rica
  5. 5.Centro Nacional de Innovaciones Biotecnológicas (CENiBiot)CeNAT-CONARESan JoseCosta Rica
  6. 6.Herbario Juvenal Valerio RodríguezUniversidad NacionalHerediaCosta Rica
  7. 7.Escuela de BiologíaInstituto Tecnológico de Costa RicaCartagoCosta Rica

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