Natural Product Research

Formerly Natural Product Letters

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Chemical composition of Helietta parvifolia and its

in vitro anticholinesterase activity

Víctor Gómez-Calvario, M. Ángeles Ramírez-Cisneros, Macdiel Acevedo-

Quiroz & Maria Yolanda Rios

To cite this article: Víctor Gómez-Calvario, M. Ángeles Ramírez-Cisneros, Macdiel Acevedo-

Quiroz & Maria Yolanda Rios (2017): Chemical composition of Helietta parvifolia and its in vitro
anticholinesterase activity, Natural Product Research, DOI: 10.1080/14786419.2017.1410808

To link to this article: https://doi.org/10.1080/14786419.2017.1410808

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Published online: 04 Dec 2017.

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 Natural Product Research, 2017
https://doi.org/10.1080/14786419.2017.1410808

SHORT COMMUNICATION

Chemical composition of Helietta parvifolia and its in vitro

anticholinesterase activity
Víctor Gómez-Calvario, M. Ángeles Ramírez-Cisneros  , Macdiel Acevedo-Quiroz and
Maria Yolanda Rios 
Downloaded by [Karolinska Institutet, University Library] at 17:32 04 December 2017

Centro de Investigaciones Químicas, IICBA, Universidad Autónoma del Estado de Morelos, Cuernavaca, México.

ABSTRACT ARTICLE HISTORY

The chemical composition of the essential oil and the n-hexane (Hex), Received 5 October 2017
Ethyl Acetate (EtOAc) and butanol (BuOH) extracts from the leaves Accepted 14 November 2017
of Helietta parvifolia were determined by detailed GC-MS analysis,
KEYWORDS
spectroscopic and spectrometric data. Eighty-four compounds were Helietta parvifolia; GC-MS;
identified, revealing a furoquinoline alkaloid-rich composition. The PCA; phytochemical analysis;
phytochemical analysis of the extracts allowed the isolation of eigth furoquinoline alkaloids; RI;
furoquinoline alkaloids. Retention indices in GC-MS for six of this anticholinesterase activity
alkaloids are reported for the first time. Furoquinoline alkaloids are
acethylcholinesterase inhibitors. Thus, the essential oil and extracts
were submitted to this in vitro assay. The EtOAc and BuOH extracts
showed potent activity, with IC50 of 9.7 and 12.9 μg mL−1, respectively.
Additionaly, a correlation of their chemical constituents, established by
principal component analysis (PCA) demostrated a similar profile and
a high content of alkaloids. It is for these reasons that we can assume
that the alkaloid content in these extracts could be responsible for
their anticholinesterase activity.

1. Introduction
Helietta genus (Rutaceae) is composed of eight species (Pirani 1998). Some of these have
exhibited antibacterial (Ferronatto et al. 2012) and antileishmanial (Ferreira et al. 2010) activ-
ities and act as adjuvants of central nervous system depressant effects (Goloubkova et al.

CONTACT  Maria Yolanda Rios  myolanda@uaem.mx

 Supplemental data for this article can be accessed at https://doi.org/10.1080/14786419.2017.1410808.
© 2017 Informa UK Limited, trading as Taylor & Francis Group
 2   V. GÓMEZ-CALVARIO ET AL.

1998). Helietta parvifolia A. Gray is an aromatic tree or shrub 4–5 m in height that grows in
the southeastern territories of the United States and the northeastern regions of México
(Maiti et al. 2016). This plant exhibits significant acute and chronic anti-inflammatory
(Marroquin-Segura et al. 2009), fungicide and insecticide activities (Anaya et al. 1987). A
preliminary study of its essential oil describes the identification of four phenylpropenes as
major constituents (Domínguez et al. 1971). Similarly, only four furoquinoline alkaloids have
been isolated from this plant: kokusaginine, flindersiamine, isoflindersiamine and heliparv-
ifoline (Domínguez et al. 1971; Chang et al. 1976). Interestingly, several alkaloids have been
associated with the inhibition of acetylcholinesterase enzyme (Houghton et al. 2006). Thus,
due to H. parvifolia’s capacity to produce furoquinoline alkaloids, it could potentially inhibit
Downloaded by [Karolinska Institutet, University Library] at 17:32 04 December 2017

this enzyme. Acetylcholinesterase inhibitors (AChEIs) have demonstrated clinical relevance

as therapeutic agents in patients with Alzheimer’s disease (AD) (Houghton et al. 2006), that
is the most frequent neurodegenerative disease worldwide (Jellinger 2006). AD is charac-
terised by a progressive cognitive dysfunction resulting in a deficiency of cholinergic activity
in the brain, causing impaired memory and senile dementia (Jellinger 2006). To date, no
study regarding the evaluation of the inhibition of acetylcholinesterase has been performed
for any of the species of the Helietta genus. Herein, the identification of chemical components
of the essential oil, and the n-hexane (Hex), Ethyl acetate (EtOAc) and Butanol (BuOH) extracts
of leaves from H. parvifolia, using GC-MS analysis, spectroscopic and spectrometric data, as
well as PCA, and its anticholinesterase activity were performed.

2.  Results and discussion

2.1.  Chemical composition
The chemical composition of H. parvifolia by GC-MS analysis is reported in Table S1. Eighty-
four compounds were identified from the essential oil and the Hex, EtOAc and BuOH extracts
from this natural source, representing 84.5–90.3% of their composition, as indicated. As seen
in Table S1, the essential oil contains mainly terpenes and phenylpropenes and the Hex
extract is rich in oxygenated sesquiterpenes, phenylpropenes, fatty acids and its methyl
esters derivatives. Additionally, the EtOAc and BuOH extracts are abundant in alkaloids.
Thirty-three compounds (90.3%) were identified in the essential oil: nine monoterpenes
(19.6%), four oxygenated monoterpenes (2.9%), two trinor-sesquiterpenes (10.8%), seven
sesquiterpenes (2.3%), eight oxygenated sesquiterpenes (40.3%), one aromatic derivative
(1.4%) and two phenylpropenes (12.9%), being elemol (23, 33.6%), (E)-isosafrole (79, 12.8%)
and pregeijerene B (15, 10.5%) the most abundant compounds. It is worth highlighting that
15 has never been identified from the Helietta genus. Regarding the Hex extract, thirty-six
compounds (84.5%) were identified: two sesquiterpenes (1.24%), ten oxygenated sesquit-
erpenes (24.3%), two fatty acids (12.7%), five fatty acid ester derivatives (18.9%), one oxy-
genated diterpene (5.8%), six alkaloids (6.1%), one sterol (1.4%), three triterpenes (3.5%) and
six phenylpropenes (10.6%). Linoleic acid (37, 10.5%), 8α-11-elemodiol (34, 7.4%), elemol
(23, 7.3%), methyl linoleate (40, 7.3%), methyl oleate (41, 6.9%), phytol (43, 5.8%) and
(E)-methyl isoeugenol (81, 5.08%) were the most abundant compounds in this extract. For
the EtOAc extract, twenty-four (87.2%) compounds were identified: three oxygenated ses-
quiterpenes (2.9%), two fatty acid ester derivatives (1.7%), one oxygenated diterpene (0.8%),
sixteen alkaloids (79.4%), one aromatic derivative (0.8%), and one phenylpropene (1.5%). It
 NATURAL PRODUCT RESEARCH   3

is important in this chemical analysis to note the high abundance of the furoquinoline alka-
loids kokusaginine (49, 33.4%) and flindersiamine (52, 20.5%). Finally, twenty-one com-
pounds (85.8%) were identified in the BuOH extract: nineteen alkaloids (79.4%) and two
aromatic derivatives (6.4%). Similarly, the contribution to the chemical composition of three
furoquinolone alkaloids kokusaginine (49, 14.3%), kokusaginine isomer (46, 13.3%) and
flindersiamine (52, 7.5%) was significant. Flash column chromatography of the Hex and
EtOAc extracts allowed the isolation of sixteen compounds. Based on their spectroscopic
and spectrometric data, these compounds were established as the sesquiterpenes 23, 25
and 34, the triterpenes 73–74, the phenylpropenes 79–81, and the eight furoquinoline
alkaloids 49, 52–53, 58, 62, 65–66 and 68 (selected spectra S1–S6). These alkaloids were
Downloaded by [Karolinska Institutet, University Library] at 17:32 04 December 2017

used as references for GC-MS to determine their RI and fragmentation patterns, providing
thus a rapid identification method for this kind of compounds (See Tables S1 and S2 sup-
plementary material).

2.2.  Anticholinesterase activity

Essential oil and extracts of H. parvifolia were tested as acetylcholinesterase inhibitors
(AChEIs) sources using Ellman’s method (Ellman et al. 1961). EtOAc and BuOH extracts inhib-
ited acetylcholinesterase (AChE) (IC50 = 9.66 and 12.88 μg mL−1, respectively, Table S2),
demonstrating equipotency when compared with other extracts reported as AChEIs
(SatheeshKumar et al. 2010). The essential oil and Hex extract showed no activity. These
results can be associated with the high content of alkaloids in the active extracts (79%),
especially in the 2:1 mixture found of the major compounds 49 and 52 (53.9 and 21.8% for
EtOAc and BuOH extracts, respectively). Additionally, compound 49 (IC50 = 11.93 μg mL−1)
(Cardoso-Lopes et al. 2010) showed to be two times more potent than 52 (28.19 μg mL−1,
Table S2).

2.3.  PCA analysis

PCA of the essential oil and the three extracts of H. parvifolia showed consistency in all
replicates for each extract type and oil. Furthermore, the similarity in the X axis between
EtOAc and BuOH extracts agrees with their anti-AChE activity (Figure S1).

3. Conclusions
EtOAc and BuOH extracts are related by PCA and contain an important amount of alkaloids
acting as AChEIs. The total chemical composition of the essential oil and the three extracts
are constituted by eighty-four compounds, being the alkaloids the most abundant. RI for
six of these alkaloids are reported for the first time to be used for their rapid identification
by GC-MS. In conclusión, H. parvifolia is an important source of furoquinoline alkaloids, and
it is a promising anticholinesterase plant with potential use in the treatment of Alzheimer’s
disease.

Acknowledgment
We are grateful to María Medina for the technical assistance.
 4   V. GÓMEZ-CALVARIO ET AL.

Disclosure statement
No potential conflict of interest was reported by the authors.

Funding
This work was supported by the CONACyT [grant number 241044]. V.G.C. was a PROMEP fellow [grant
number 10944].

ORCID
Downloaded by [Karolinska Institutet, University Library] at 17:32 04 December 2017

M. Ángeles Ramírez-Cisneros   http://orcid.org/0000-0003-4696-7359

Maria Yolanda Rios   http://orcid.org/0000-0002-8875-8734

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