Research Article
Chris Packer*
Chris Packer*
Corresponding Author
D. Gary Young Research Institute, Lehi, UT 84043, USA.
E-mail: cpacker@youngliving.com, Tel: +1 208 5300067
Adrian Abad
Adrian Abad
Finca Botanica Aromatica, Guayaquil, 090151, EC, Ecuador.
E-mail: adabad@youngliving.com
Tulio Orellana
Tulio Orellana
Finca
Botanica Aromatica, Guayaquil, 090151, EC, Ecuador.
Tyler M. Wilson
Tyler M. Wilson
D.
Gary Young Research Institute, Lehi, UT 84043, USA.
Nadia Cedeño
Nadia Cedeño
Independent
researcher, UT, USA
Eugenio Caruajulca
Eugenio Caruajulca
Finca
Botanica Aromatica, Guayaquil, 090151, EC, Ecuador.
Orlando Pacheco
Orlando Pacheco
Finca
Botanica Aromatica, Guayaquil, 090151, EC, Ecuador.
Abstract
The essential oil from
the leaves of Siparuna lepidota (Kunth) A. DC. was collected from
cultivated species in Guayaquil, Ecuador. The oil was obtained by steam
distillation and analyzed using GC/MS and GC/FID and it revealed a high content
of monoterpene hydrocarbons (83.7%). The essential oil was rich in limonene
(71.5%), with other prominent compounds such as β-pinene (5.2%), α-pinene
(2.8%), myrcene (1.7%), α-terpinolene (2.3%), δ-cadinene (2.1%), α-copaene
(1.7%), and (E)-cadina-1,4-diene (1.4%). To the best of the authors' knowledge,
this study is the first to present the chemical composition of Siparuna
lepidota essential oil, providing fundamental data for future research into
its ethnobotanical uses and biological properties.
Keywords
Chemical profile, Ecuador, essential oil, GC/FID,
GC/MS, Siparuna lepidota, steam distillation.
The Siparunaceae family contains two genera: Glossocalyx
and Siparuna. While Glossocalyx is comprised of only a few
species in West Africa, Siparuna has about 58 species with a range that
includes Central America and the West Indies and reaches throughout northern
South America to Paraguay [1-3]. Most of
these species are shrubs, with some trees, and are found at elevations from
sea-level to 3800 m and in a wide variety of habitats, spanning lowland rain
forest, montane forests, subpáramo scrub forest, lower elfin forest, and
grassland gallery forest [1].
Some species of the Siparuna genus are noted
for their characteristic, strong, citrus aroma given off by the leaves, fruit,
and bark [1, 4]. Siparuna spp. are
notable not only for their large and abundant oil-containing cells in each of
their plant parts but also for their rich composition of biologically active
compounds, including benzylisoquinoline alkaloids, sesquiterpenes, and
flavonoids [1, 4]. Siparuna species
have been traditionally utilized by native peoples throughout South and Central
America for various medicinal purposes. They are used in traditional medicine
as antimalarials and febrifuges, with practices including the boiling of
macerated leaves to create medicinal baths aimed at alleviating fever, cold
symptoms, and rheumatism [1, 5]. The research
also highlighted the extracts from the leaves of S. grandiflora
(referred to as S. tonduziana), S. pauciflora, and S.
thecaphora demonstrate activities against the malaria parasite, which
supports their use in traditional antimalarial therapies [1]. Additionally, applications of Siparuna spp.
leaves or bark in poultices for the treatment of snake bites and minor wounds
are documented, as is the use by the Ecuadorian Quichua of heated bark from
species such as S. sessiliflora and S. thecaphora for hastening
the recovery from herpes sores [1].
Ecuador, a country with some of the greatest
biodiversity in the world [6], has
approximately 40 species of the Siparuna genus [7],
and Siparuna lepidota is one such species. S. lepidota is
a shrub or treelet that can be 3-10 m in height, with yellow flowers, and small
round fruit that is reddish-purple when ripe [1]. Common
names include chiri guayusa, guayusa de montaña, and limoncello [1, 8]. While the ethnobotanical and medicinal
uses of Siparuna lepidota are not extensively documented, it was
reported that the fruits were decocted in water to produce an extract used for
treating stomach colics, in medicinal baths, and to impart a lemon flavor to
beverages [1]. Additionally, the juice
extracted from the leaves of this species is applied topically as a remedy for
ear pain [8].
A few species of Siparuna, such as S. echinata, S. guianensis, S. muricata, S. thecaphora, S. cymosa, and S. brasiliensis have been studied for their essential oil content [9-15]. Little research exists to elucidate the chemical properties of Siparuna lepidota essential oil, which may substantiate its traditional uses. To the authors’ best knowledge, the essential oil of S. lepidota has not been previously reported. In the present study, GC/MS and GC/FID analytical techniques are used to establish the chemical profile of this essential oil from cultivated samples in Guayaquil, Ecuador, providing a foundation for future research.
Siparuna lepidota leaves (Fig. 1) were collected in August 2023 from cultivated populations in Guayaquil, Ecuador (2°16'36.8"S 80°03'56.1"W). Plant material was stored in a shaded location for 2 days before distillation. A representative voucher sample of S. lepidota was deposited in the herbarium Universidad de Guayaquil (13.553GUAY). Steam distillation was carried out for 2 hours. The essential oil obtained was separated by a cooled condenser, collected, filtered, and stored in sealed amber vials at room temperature (25 °C) until analysis. The essential oil yield was calculated as the ratio of the essential oil volume (mL) to the plant material mass (kg) before the distillation process.
Figure 1. Botanical
illustration of Siparuna lepidota plant part used in the study, namely
the leaves. Illustrated by Rick Simonson, Science Lab Studios, Inc. (Kearney,
NE, USA).
Essential oil was analyzed, and volatile compounds were identified by GC/MS using an Agilent 7890BGC/5977B MSD (Agilent Technologies, Santa Clara, CA, USA) and Agilent J&W DB-5, 60 m × 0.25 mm,0.25 μm film thickness, fused silica capillary column. Operating conditions: 0.2 μL of the sample, 25:1 split ratio, initial oven temperature of 60 °C with an initial hold time of 2 min, oven ramp rate of 4.0 °C per minute to 245 °C with a hold time of 5 min, helium carrier gas. The electron ionization energy was 70 eV, scan range 35–550 amu, scan rate 2.4 scans per second, source temperature 230 °C, and quadrupole temperature 150 °C. Volatile compounds were identified using the Adams volatile oil library [16] using Chemstation library search in conjunction with retention indices. Volatile compounds were quantified and reported as a relative area percent by GC/FID using an Agilent 7890B and Agilent J&WDB-5, 60 m × 0.25 mm, 0.25 μm film thickness, fused silica capillary column. Operating conditions: 0.1 μL of sample, 25:1 split injection, initial oven temperature at 40 °C with an initial hold time of 2 min, oven ramp rate of 3.0 °C per minute to 250 °C with a hold time of 3 min, helium carrier gas. Essential oil samples were analyzed in triplicate by GC/FID to ensure repeatability (standard deviation < 1 for all compounds).
3.
Results and discussion
The essential oil yield of Siparuna lepidota was 13.3 mL/kg, and the chemical profile is detailed in Table 1, revealing this essential oil is rich in monoterpene hydrocarbons (83.7%). Twenty-seven compounds of S. lepidota essential oil were identified. Limonene was the most abundant component in the essential oil (71.5%). Other notable monoterpenes include α-pinene (2.8%), β-pinene (5.2%), myrcene (1.7%), and α-terpinolene (2.3%), The second most abundant group in the essential oil were the sesquiterpene hydrocarbons, including compounds such as δ-cadinene (2.1%), α-copaene (1.7%), and (E)-cadina-1,4-diene (1.4%).
Table 1. Chemical profile of S. lepidota essential oil determined by GC/FID.
KI |
Compound Name |
Area percentage (%) |
932 |
α-Pinene |
2.8 |
969 |
Sabinene |
0.2 |
974 |
β-Pinene |
5.2 |
Myrcene |
1.7 |
|
1024 |
Limonene |
71.5 |
1086 |
α-Terpinolene |
2.3 |
1137 |
(E)-Limonene
oxide |
0.3 |
1179 |
p-Cymen-8-ol |
0.1 |
1186 |
α-Terpineol |
0.1 |
1235 |
Neral |
0.2 |
1249 |
Geraniol |
0.2 |
1264 |
Geranial |
0.3 |
1348 |
1.1 |
|
1374 |
α-Copaene |
1.7 |
1387 |
β-Cubebene |
1.3 |
1417 |
(E)-Caryophyllene |
0.7 |
1448 |
cis-Muurola-3,5-diene |
1.3 |
1452 |
α-Humulene |
0.6 |
1458 |
Allo-aromadendrene |
0.5 |
1478 |
γ-Muurolene |
0.3 |
1480 |
Germacrene
D |
1.1 |
1493 |
(E)-Muurola-4(14),5-diene |
0.9 |
1500 |
Bicyclogermacrene |
0.5 |
1521 |
(E)-Calamenene |
0.5 |
1522 |
δ-Cadinene |
2.1 |
1533 |
(E)-Cadina-1,4-diene |
1.4 |
|
Compound
Classes |
|
|
Monoterpene hydrocarbons |
83.7 |
|
Oxygenated monoterpenes |
1.2 |
|
Sesquiterpene hydrocarbons |
14.0 |
|
Total
identified compounds |
98.9 |
Note: Essential oil sample was analyzed in triplicate to ensure repeatability
(standard deviation < 1). Unidentified compounds of less than 0.5% are not
included. KI is the Kovat’s Index previously calculated by Robert Adams
using a linear calculation on a DB-5 column [16]. Relative area percent was
determined by GC/FID.
Reviewing the chemical compositions of essential oils from species within the same genus, previous studies have demonstrated that the essential oil of S. echinata is characterized by a high content of monoterpene hydrocarbons, with α-pinene, β-pinene, β-myrcene, and limonene being the major compounds, the latter constituting 10.0% [9]. Similarly, the essential oil of S. muricata is dominated by hydrocarbon monoterpenes, with α-pinene as the principal component and a significant amount of limonene (8.7%) [7]. In contrast to the previously mentioned species, which is characterized by major monoterpenes, the essential oil of S. guianensis leaves has been reported to contain major compounds such as the sesquiterpenes γ-muurolene, curzerene, and curzerenone [10]. Furthermore, distinct chemotypes of S. guianensis have been identified including one with α-bisabolol as the major compound, and the other with germacrene D as the predominant component [11]. For S. thecaphora, the major compounds identified in its essential oil are spathulenol, and 2-tridecanone [12]. Additionally, the essential oil of S. cymosa is primarily composed of α-bisabolol [14], while S. brasiliensis is characterized by γ-muurolene and 2-undecanone as its major components [15].
Comparing the chemical composition of Siparuna lepidota essential oil with previously reported species within the genus Siparuna, both similarities and notable differences are observed. In our study, limonene was the most abundant component (71.5%), being significantly higher than the limonene content reported in S. muricata (8.7%) and in S. echinate (10.0%) [7, 9]. Additionally, while S. lepidota essential oil contains α-pinene, β-pinene, and myrcene, these monoterpenes are also present in S. echinata and S. muricata, though in different proportions, highlighting the unique profile of S. lepidota species essential oil, which is particularly rich in limonene. These differences in chemical composition not only underscore the chemical diversity within the genus Siparuna, but also could have significant implications for their uses in the fragrance and aromatherapy industries, where limonene-rich essential oils are valued for their citrus-like properties and potential therapeutic benefits.
The most
abundant chemical constituents of essential oils generally dictate their
bioactivities [17, 18]. Limonene, the most
abundant compound present in the essential oil of S. lepidota, has been
studied for its biological activities. Limonene has been shown to have
antiproliferative, apoptotic, and anti-carcinogenic properties
[19, 20]. Many studies have revealed that
limonene provides antioxidant properties [21].
Also, this compound has been reported as an effective anti-inflammatory [21], which could be related to the traditional
use of S. lepidota as a remedy for ear pain. Future studies need to be
conducted to investigate the biological activities of S. lepidota essential
oil.
4.
Conclusions
To the best of
the authors' knowledge, this study is the first to present the chemical
composition of Siparuna lepidota essential oil. The essential oil of S.
lepidota, with a high limonene concentration (71.5%), indicates significant
potential for therapeutic and commercial applications. Although it shares some
monoterpenes with other Siparuna species, the high limonene content is
particularly notable. This suggests antioxidant and anti-inflammatory benefits,
consistent with traditional use, and highlights potential applications in the pharmaceutical
and cosmetic industries. Further studies are recommended to evaluate its
biological activities and explore industrial and medicinal applications.
Authors’ contributions
Conceptualization,
C.P.; Methodology, C.P., A.A.; Software, C.P., A.A., T.O.; Validation, C.P.;
Formal analysis (GC/MS, GC/FID), C.P., A.A., T.M.W., T.O.; Investigation, C.P.,
A.A.; Resources, C.P., N.C., E.C., O.P.; Data curation, C.P., A.A.; Writing–
original draft, C.P. A.A.; Writing–review & editing, C.P., A.A., N.C., T.O.,
E.C., O.P., T.M.W.
Acknowledgements
The authors want to thank the D. Gary Young Research
Institute and Finca Botanica Aromatica, for providing support for this project.
Appreciation would also like to be extended to Rick Simonson (Science Lab
Studios) for the botanical illustration.
Funding
This research was funded by Young Living Essential
Oils.
Availability of data and
materials
All data will be made available on
request according to the journal policy.
Conflicts of interest
The authors declare no conflict of interest. The
funding entity had no role in the design of the study, in the collection,
analysis, or interpretation of data, in the writing of the manuscript, or in
the decision to publish the results.
References
1.
Renner, S.; Gerlinde, H. “Siparunaceae.” Flora Neotropica. 2005, 95.
2.
Christenhusz
M.; Byng J. The number of known plant species in the world and its annual
increase. Phytotaxa. 2016, 261 (3), 201-217. https://doi.org/10.11646/phytotaxa.261.3.1.
3.
WFO Plant
List. Siparunaceae (A.DC.) Schodde.
4.
Silva, I.; Oliveira, F.; Oliveira, R. Siparuna Aublet genus
(Siparunaceae): from folk medicine to chemical composition and biological
activity. Trends Phytochem. Res. 2021, 5(4), 168-189.
https://doi.org/10.30495/tpr.2021.1934361.1211.
5.
Leal, C.; Simas R.; Miranda, M.; Campos, M.; Gomes, B.; Siqueira, M.; Vale,
G.; Almeida, C.; Leitão, S.; Leitão, G. Amazonian Siparuna extracts as
potential anti-influenza agents: metabolic fingerprinting. J. Eth. Pharm. 2021,
270, 113788. https://doi.org/10.1016/j.jep.2021.113788.
6.
Jorgensen, P.; Leon-Yanez, S. Catalogue of the Vascular Plants of
Ecuador, Missouri Botanical Garden Press, St. Louis, Mo, USA, 1999.
7.
Morocho, V.; Hidalgo, M.; Delgado, I.; Cartuche, L.; Cumbicus, N.;
Valarezo, E. Chemical composition and biological activity of essential oil from
leaves and fruits of Limoncillo (Siparuna muricata (Ruiz & Pav.) A.
DC.). Antibiotics. 2023, 12(1), 82. https://doi.org/10.3390/antibiotics12010082.
8.
Ballesteros, J.; Bracco, F.; Cerna, M.; Finzi, P.; Vidari, G.
Ethnobotanical research at the Kutukú Scientific Station, Morona-Santiago,
Ecuador. BioMed. Res. Int. 2016, 2016, 1-18.
https://doi.org/10.1155/2016/9105746.
9.
García, J.; Gilardoni, G.; Cumbicus, N.; Morocho, V. Chemical analysis of the essential oil from Siparuna echinata (Kunth)
A. DC. (Siparunaceae) of Ecuador and isolation of the rare terpenoid sipaucin A. Plants. 2020, 9(2), 187. https://doi.org/10.3390/plants9020187.
10.
Souza, J.; Silva, L.; Romano, C.; Cunha, L.; Oliveira, J.; Borges, L.;
Sousa, T.; Paula, J. Chemical composition and seasonal variation of the
volatile oils from Siparuna guianensis Aubl. leaves collected from Monte
do Carmo, Tocantins. RSD. 2022, 11(1), e30011124908.
https://doi.org/10.33448/rsd-v11i1.24908.
11.
Diniz, J.; Marchesini, P.; Zeringóta, V.; Matos, S.; Novato, T.; Melo,
D.; Vale L.; Lopes, W.; Gomes, G.; Monteiro, C. Chemical composition of
essential oils of different Siparuna guianensis chemotypes and
their acaricidal activity against Rhipicephalus microplus (Acari:
Ixodidae): influence of α-bisabolol. Int. J. Acarol. 2021,
48(1), 36-42. https://doi.org/10.1080/01647954.2021.2009910.
12.
Melo, D.; Miranda, M.; Junior, W.; Alcoba, A.; Andrade, P.; Silva, T.;
Cazal, C.; Martins, C. Anticariogenic and antimycobacterial activities of the
essential oil of Siparuna guianensis Aublet (Siparunaceae). Orbital.
2017, 9(1), 55-60. http://dx.doi.org/10.17807/orbital.v0i0.930.
13.
Vila, R.; Iglesias, J.; Cañigueral, S.; Santana, A.; Solís, P.; Gupta,
M. Chemical composition and biological activity of the leaf oil of Siparuna
thecaphora (Poepp. et Endl.) A. DC. J. Essent. Oil Res. 2022, 14(1), 66-67.
https://doi.org/10.1080/10412905.2002.9699767.
14.
da Silva, R.; Evangelista, F.; Sabino, A.; da Silva, L.; de Oliveira,
F.; de Oliveira, R. Cytotoxicity assessment of Siparuna cymosa essential
oil in the presence of myeloid leukemia cells. Rev. Virtual Quim. 2020, 12,
1381-1388. https://doi.org/10.21577/1984-6835.20200109.
15.
Gonçalves, A.; Sabino, A.; de Oliveira,
F.; de Oliveira, R. Perfil Químico dos Óleos Essenciais das Folhas e Caules de Siparuna
brasiliensis (Spreng.) A. DC. Rev. Virtual Quim. 2023, 15(4), 706-712.
https://doi.org/10.21577/1984-6835.20230002.
16.
Adams, R.P. Identification of essential oil components by gas
chromatography/mass spectrometry, 4th edn.; Allured Publ.: Carol
Stream, IL, USA, 2007.
17.
Pavela, R. Essential oils for the development of eco-friendly mosquito
larvicides: a review. Ind. Crops Prod. 2015, 76(2015), 174-187. https://doi.org/10.1016/j.indcrop.2015.06.050.
18.
Haro, J.; Castillo, G.; Martínez, M.; Espinosa, H. Clove essential oil (Syzygium
aromaticum L. Myrtaceae): Extraction, chemical composition, food
applications, and essential bioactivity for human health. Molecules. 2021,
26(21), 6387. https://doi.org/10.3390/molecules26216387.
19.
Mohammed, M.; Babeanu, N.; Cornea, C.; Radu, N. Limonene- A biomolecule
with potential applications in regenerative medicine. Scientific Bulletin
Series F. Biotechnologies. 2022, 26(2), 139-150.
20. Chen, X.; Ding, Y.; Guan, H.; Zhou, C.; He, X.; Shao, Y.; Wang, Y.; Wang, N.; Li, B.; Lv, G.; Chen, S. Pharmacological effects and potential applications of limonene from citrus plants: A review. Nat. Prod. Commun. 2024, 19(5), 1-12. https://doi.org/10.1177/1934578X241254229.
21. Anandakumar, P.; Kamaraj, S.; Vanitha, M. D‐limonene: A multifunctional compound with potent therapeutic effects. J. Food Biochem. 2020, 45(1), e13566. https://doi.org/10.1111/jfbc.13566.

This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).
Abstract
The essential oil from
the leaves of Siparuna lepidota (Kunth) A. DC. was collected from
cultivated species in Guayaquil, Ecuador. The oil was obtained by steam
distillation and analyzed using GC/MS and GC/FID and it revealed a high content
of monoterpene hydrocarbons (83.7%). The essential oil was rich in limonene
(71.5%), with other prominent compounds such as β-pinene (5.2%), α-pinene
(2.8%), myrcene (1.7%), α-terpinolene (2.3%), δ-cadinene (2.1%), α-copaene
(1.7%), and (E)-cadina-1,4-diene (1.4%). To the best of the authors' knowledge,
this study is the first to present the chemical composition of Siparuna
lepidota essential oil, providing fundamental data for future research into
its ethnobotanical uses and biological properties.
Abstract Keywords
Chemical profile, Ecuador, essential oil, GC/FID,
GC/MS, Siparuna lepidota, steam distillation.

This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).

Editor-in-Chief

This work is licensed under the
Creative Commons Attribution 4.0
License.(CC BY-NC 4.0).