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, USA
E mail: adabad@youngliving.com
Tyler M. Wilson
Tyler M. Wilson
D. Gary Young
Research Institute, Lehi, UT 84043, USA
Eugenio Caruajulca
Eugenio Caruajulca
Finca Botanica
Aromatica, Guayaquil, 090151, EC, USA
Tulio Orellana
Tulio Orellana
Finca Botanica
Aromatica, Guayaquil, 090151, EC, USA
Orlando Pacheco
Orlando Pacheco
Finca Botanica
Aromatica, Guayaquil, 090151, EC, USA
Abstract
Cyperus luzulae (L.) Rottb. ex Retz
is an understudied species within the Cyperus genus. To the best of the authors’ knowledge, the
essential oil from this species has never been fully characterized. Essential
oil obtained through steam distillation of C.
luzulae rhizomes was analyzed to
establish the chemical profile by GC/FID and GC/MS. The chemical profile
revealed a high content of sesquiterpene hydrocarbons (36.9%) and oxygenated
sesquiterpenes (34.9%). The major compounds of the oil were caryophyllene oxide
(23.5%),
β-selinene (12.9%), and α-copaene (8.3%).
These results provide fundamental data for future investigations into the
ethnobotanical and pharmacological uses of this species, broadening the scope
of knowledge on the Cyperus genus and its potential applications.
Abstract Keywords
Chemical profile,
essential oil, ethnobotanical, Cyperaceae, Cyperus
luzulae, gas chromatography,
sesquiterpene.
1. Introduction
The Cyperaceae family is comprised
of annual herbs and rhizomatous perennials, and covers a broad array of
diversity within the herbaceous plant kingdom encompassing around 5700 unique
species [1]. These species are distributed
among two distinct subfamilies, which are further divided into 24 tribes and
classified under 95 different genera [1, 2]. The
broad geographical reach of the plant family provides a vast array of diversity
in their biological traits and habits [3].
Cyperus luzulae (L.) Rottb. ex Retz, commonly known as “Piri Piri”, is one of the members of the Cyperaceae family
growing primarily in tropical and subtropical regions of America that has
cespitose rhizomes, brown to reddish leaf bases, and it has been traditionally
used for a variety of social and/or medicinal purposes [4,
5].
The Cyperus luzulae plant holds historical and therapeutic
significance across various cultures. Historically, the Shipibo-Conibo peoples
utilized the plant's rhizomes as an aphrodisiac, while in Guatemala, its
crushed rhizome forms the basis of the Mesoamerican drink, "atole" [5, 6]. Indigenous stories from the Huaorani group
in San Francisco, Ucayali, Peru, tell of shamans crafting love potions from C. luzulae to attract partners [7].
Furthermore, conversations with a Shaman from the Shipibo-Konibo
highlight its use in childbirth, for its pain-minimizing effects, and as a
diuretic aiding individuals with fluid retention or high blood pressure [8]. In Panama, it has therapeutic applications
for eye infections and complications during childbirth [9].
While this is an
understudied plant species, recent research has been done to investigate
botanical, ecology, and utility of this species [10–12].
Scientific research underscores its medicinal potential. Methanol
extracts from C.
luzulae roots have exhibited DNA
intercalation, tumor inhibition, and cytotoxic effects in V79 cultures, hinting
at its oncological applications [13]. Both
aqueous and methanol extracts of the plant also demonstrate molluscicidal
activity against Biomphalaria
glabrata, a known host of
schistosomiasis [14].
Despite the
referenced studies, to the best of the authors’ knowledge, the chemical
composition of the essential oil of C.
luzulae rhizomes has not been
reported. This lack of knowledge limits the understanding of its potential
usefulness in terms of possible applications. This study aims to determine the
chemical composition of the essential oil of rhizomes from C. luzulae to provide fundamental data
for future investigations of this species.
2. Materials and methods
Cyperus
luzulae rhizomes were collected
in December 2022 from cultivated populations in Coronel Portillo province, Peru
(8°16'26.1" S 74°38'40.1" W). The rhizomes were washed with water to
remove the soil surrounding them and were left to dry out of sunlight for 4
days. Then the rhizomes were crushed and weighed, and the extraction process
was conducted. A representative voucher sample of the species is held at the
Universidad Nacional de Cajamarca (Herbario Isidoro Sánchez Vega_UNC; herbarium
code CPUN).
The distillation was conducted in a 250 L
distillation chamber (Albrigi Luigi S.R.L., Italy). Distillation was conducted
by steam distillation for 4 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.
The essential oil compounds were analyzed and
identified by GC/MS using Agilent7890B GC/5977B MSD (Agilent Technologies,
Santa Clara, CA, USA) and Agilent J&W DB-5, 0.25 mm x 60m, 0.25 μm film
thickness, fused silica capillary column. Operating conditions: 0.1 μL of the sample
(20% soln. for essential oils in ethanol), 100:1 split ratio, initial oven
temp. of 40 °C with an initial hold time of 5 min., oven ramp rate of 4.5 °C
per min. to 310 °C with a hold time of 5 min. The electron ionization energy
was 70 eV, scan range 35-650 amu, scan rate2.4 scans per sec., source temp. 230
°C, and quadrupole temp. 150 °C. Volatile compounds were identified using the
Adams volatile oil library [15] using
Chemstation library search in conjunction with retention indices. Volatile compounds
were quantified and are reported as a relative area percent by GC/FID using an
Agilent7890B GC and Agilent J&W DB-5, 0.25 mm x 60 m, 0.25μm film
thickness, fused silica capillary column. Operating conditions: 0.1 μL of
sample (20% soln. for essential oil in ethanol, 1% for reference compounds in
ethanol, 0.1% soln. for C7-C30 alkanes in hexane), 25:1 split ratio, initial
oven temp. of 40 °C with an initial hold time of 2 min., oven ramp rate of 3.0
°C per min. to 250 °C with a hold time of 3 min. The essential oil sample was
analyzed in triplicate by GC/FID to ensure repeatability (standard deviation
< 1 for all compounds). Compounds were assigned using retention indices
coupled with the retention time data of reference compounds (MilliporeSigma,
Sigma-Aldrich, St. Louis, MO, USA).
3.
Results and discussion
The essential oil yield from rhizomes of Cyperus
luzulae was 2.1 mL/kg, and the chemical profile is detailed in Table 1,
revealing that this essential oil is rich in
sesquiterpene hydrocarbons and oxygenated sesquiterpenes.
Table
1.
Chemical profile of Cyperus luzulae rhizome essential oil determined by
GC/FID.
KI |
Compound Name |
Area
percentage (%) |
932 |
α-Pinene |
1.1 |
974 |
β-Pinene |
1.8 |
1020 |
p-Cymene |
0.2 |
Limonene |
0.6 |
|
1026 |
1,8-cineole |
0.8 |
1135 |
Nopinone |
0.2 |
1135 |
1.8 |
|
1174 |
Terpinen-4-ol |
0.2 |
1186 |
α-Terpineol |
0.4 |
1194 |
Myrtenol |
2.0 |
1204 |
Verbenone |
0.3 |
1369 |
Cyclosativene |
1.2 |
1374 |
8.3 |
|
1389 |
β-Elemene |
0.4 |
1390 |
Sativene |
0.2 |
1398 |
Cyperene |
5.9 |
1457 |
Rotundene |
1.3 |
1473* |
Unknown |
1.0 |
1478 |
γ-Muurolene |
0.6 |
1489 |
12.9 |
|
1498 |
α-Selinene |
2.1 |
1506* |
Unknown |
1.1 |
1528 |
cis-Calamene |
1.1 |
1539* |
Unknown |
0.5 |
1544 |
α-Calacorene |
1.2 |
1564 |
β-Calacorene |
1.9 |
1570* |
Unknown |
0.6 |
1582 |
23.5 |
|
1608 |
Humulene
epoxide II |
3.4 |
1620* |
Unknown |
0.5 |
1623* |
Unknown |
1.7 |
1635* |
Unknown |
1.7 |
1638* |
Unknown |
0.7 |
1643* |
Unknown |
1.5 |
1646* |
Unknown |
0.6 |
1649 |
Longiverbenone |
2.8 |
1650* |
Unknown |
1.3 |
1663* |
Unknown |
0.9 |
1666* |
Unknown |
1.1 |
1673* |
Unknown |
0.6 |
1695* |
α-Cyperone |
5.2 |
|
Other |
0.2 |
|
Total
(%) |
81.2 |
Compound Classes |
|
|
Monoterpene hydrocarbons |
3.7 |
|
Oxygenated monoterpenes |
5.5 |
|
Sesquiterpene hydrocarbons |
36.9 |
|
Oxygenated sesquiterpenes |
34.9 |
|
Note: Essential oil sample was analyzed in
triplicate to ensure repeatability (standard deviation < 1 for all
values). 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 [15]. *KI not previously
calculated [15] and manual calculations
performed using alkane standards. |
Twenty-seven compounds of Cyperus
luzulae essential oil were identified in this study. The primary monoterpene
hydrocarbons were α-pinene (1.1%) and β-pinene (1.8%). The major oxygenated
monoterpenes were (E)-pinocarveol (1.8%) and myrtenol (2.0%). The principal
sesquiterpene hydrocarbons were β-selinene (12.9%), α-copaene (8.3%), cyperene
(5.9%), and α-selinene (2.1%). The most abundant oxygenated sesquiterpenes were
caryophyllene oxide (23.5%), α-cyperone (5.2%), humulene epoxide II (3.4%), and
longiverbenone (2.8%). Figure 1 is provided for a more intuitive visual
representation.
Figure 1.
Comparison of compound concentrations in the essential oil of Cyperus
luzulae.
Comparing our results with species
of the same genus, Cyperus rotundus, a commonly studied species in the
same genus with rhizome-extracted essential oil, shows a high concentration of cyperene,
α-cyperone, rotundene, α-selinene, β-selinene [16-19].
The essential oil of Cyperus distans is characterized by
prominent compounds such as cyperene, caryophyllene oxide, and β-pinene [20]. Additionally, Cyperus esculantus
essential oil has been documented to possess a significant concentration of
caryophyllene oxide while Cyperus spapyrus essential oil has cyperene
and copaene as principal compounds [21]. It
is noteworthy that all these compounds, known to be present in these related
species' essential oils, are also found within the C. luzulae essential
oil obtained from the current study. Despite these similarities, it's essential
to highlight that the unique combinations and concentrations of these compounds
in C. luzulae may confer this species with a unique set of properties.
Further investigations are required to explore these potential applications.
The principal chemical constituents
of essential oils generally dictate their bioactivities [22, 23]. Caryophyllene oxide is the major compound present in
the essential oil of rhizomes of Cyperus luzulae from this study. Literature
reported caryophyllene oxide affects the growth of tumor cells, and may be used
against cancer of the prostate, human breast, cancer cells of human
osteosarcoma MG-63, lung cancer cells, and gastric cancer cells [24-29], which makes C. luzulae essential
oil interesting for future studies against cancer cells.
Figure 2. Caryophyllene oxide chemical structure. Obtained from NIST [30]
Using the available mass spectral
libraries (NIST 2020) and Adams library for this study, we were only able to
identify 81.2% of Cyperus luzulae essential oil from the rhizomes. The
limitations of the analytical techniques may have hindered certain compound identification.
Advanced analytical techniques such as two-dimensional gas chromatography
coupled with time-of-flight mass spectrometry (GCxGC-TOFMS), nuclear magnetic
resonance spectroscopy (NMR), and high-resolution mass spectrometry (HRMS)
could improve identification [31]. Additionally,
the complexity of the essential oil matrix might have contributed to the
non-identification and some unidentified compounds are uncommon, suggesting
potential for future discoveries.
This
study, to the best of the authors’ knowledge, is the first to elucidate the
chemical composition of Cyperus luzulae essential oil derived from
rhizomes, with caryophyllene oxide, β-selinene, and α-copaene identified as the
predominant compounds. Despite the growing body of literature on the chemical
composition of essential oils within the Cyperus genus, there has been
limited analytical research specifically on this species. This research not
only fills a gap in our understanding but also highlights the potential
applications based on its unique chemical profile. Looking ahead, it would be
beneficial to delve deeper into the properties and applications of this
essential oil, possibly exploring its therapeutic or industrial potential. Future
research should also prioritize employing advanced analytical methods and
developing comprehensive databases for reference standards. Such endeavors will
not only facilitate the identification of unknown compounds but will also pave
the way for a more holistic understanding of this essential oil's properties
and applications.
Authors’ contributions
Conceptualization, C.P.;
Methodology, C.P. and A.A.; Software, C.P., A.A., T.M.W., and T.O.; Validation,
C.P.; Formal Analysis (GC/MS, GC/FID), C.P., A.A, T.M.W., and T.O.;
Investigation, C.P. and A.A.; Resources, C.P., E.C.; Data Curation, C.P. and
A.A.; Writing – Original Draft, C.P. and A.A.; Writing – Review & Editing,
C.P., A.A., T.M.W., T.O., E.C., and O.P.
Acknowledgements
The authors want to thank the D.
Gary Young Research Institute and Finca Botanica Aromatica, for providing
support for this project.
Funding
This
research was funded by Young Living Essential Oils
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.
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This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).
Abstract
Cyperus luzulae (L.) Rottb. ex Retz
is an understudied species within the Cyperus genus. To the best of the authors’ knowledge, the
essential oil from this species has never been fully characterized. Essential
oil obtained through steam distillation of C.
luzulae rhizomes was analyzed to
establish the chemical profile by GC/FID and GC/MS. The chemical profile
revealed a high content of sesquiterpene hydrocarbons (36.9%) and oxygenated
sesquiterpenes (34.9%). The major compounds of the oil were caryophyllene oxide
(23.5%),
β-selinene (12.9%), and α-copaene (8.3%).
These results provide fundamental data for future investigations into the
ethnobotanical and pharmacological uses of this species, broadening the scope
of knowledge on the Cyperus genus and its potential applications.
Abstract Keywords
Chemical profile,
essential oil, ethnobotanical, Cyperaceae, Cyperus
luzulae, gas chromatography,
sesquiterpene.
This work is licensed under the
Creative Commons Attribution
4.0
License (CC BY-NC 4.0).
Editor-in-Chief
Prof. Dr. Radosław Kowalski
This work is licensed under the
Creative Commons Attribution 4.0
License.(CC BY-NC 4.0).