Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)

ISSN 2616-0692 (Electronic)

Tropical Journal of Natural Product Research

Available online at https://www.tjnpr.org
Original Research Article
Phytochemical Analysis and Evaluation of the Antioxidant Activity of Cedrus atlantica
(Endl.) G. Manetti ex Carrière Stem Extracts
Mouna Ameggouz1,2*, Soufiane Drioua1, Otman El-Guourrami1, Hanane Azalmad1, Mohamed Ouajdi2, Ahmed Zahidi3, Anass
Doukkali1, Badr Satani2, Hanane Benzeid1
1
Laboratory of Analytical Chemistry, Faculty of Medicine and Pharmacy, Mohammed V University, Rabat, Morocco.
2
Chemistry and Microbiology Laboratories, Forest Research Center, Avenue Omar Ibn El Khattab, BP 763, Agdal, Morocco.
3
Laboratory of Medicinal Chemistry, Department of Drug Sciences, Faculty of Medicine and Pharmacy, Mohammed V University in Rabat, Morocco.

ARTICLE INFO ABSTRACT

Article history: Cedrus atlantica is a Mediterranean medicinal plant used to treat a number of ailments including
Received 15 February 2024 cancer and urinary tract infections. This study is aimed to analyze the phytochemical constituents
Revised 25 March 2024 and evaluates the antioxidant activity of Cedrus atlantica stem extracts. Powdered stem of Cedrus
Accepted 26 March 2024 atlantica underwent successive extraction with cyclohexane, ethyl acetate, and ethanol using
Published online 01 April 2024 Soxhlet apparatus to obtain cyclohexane (F1), ethyl acetate (F2), and ethanol (F3) extracts. The
marc was macerated with water to obtain the aqueous extract (F4). Phytochemical screening was
performed following standard procedures. Total phenolic, flavonoid, flavonol, and tannin contents
were also determined. The extracts were subjected to gas chromatography mass spectrometry
analysis. The antioxidant activity was evaluated using the DPPH, ABTS, FRAP and TAC assays.
Phytochemical screening revealed the presence of flavonoids, anthocyanins, tannins, quinines,
coumarins, terpenoids, anthraquinones, sterols, and saponins in the plant. F3 showed the highest
Copyright: © 2024 Ameggouz et al. This is an open- content of total polyphenols (237.23±1.61 mgGAE/g extract) and tannins (189.4±0.76 mgCE/g
access article distributed under the terms of the extract), while F2 exhibited the highest content of total flavonoids (81.53±1.13 mgQE/g extract)
Creative Commons Attribution License, which and flavonols (33.4±0.16 mgQE/g extract). F3 also demonstrated the most potent antioxidant
permits unrestricted use, distribution, and reproduction efficacy in the DPPH (IC50 = 19.40±0.01 μg/mL), ABTS (IC50 = 21.24±0.002 μg/mL), FRAP
in any medium, provided the original author and (EC50 = 94.36±2.51 μg/mL), and TAC (780.97±4.86 µg AAE/g extract) assays. GC-MS analysis
source are credited. identified acids, phenolic compounds, terpenes, and steroids in the extracts, suggesting their
contribution to the antioxidant activity exhibited by the plant extracts. These observations
underscore the therapeutic potential of the plant and justify its traditional medicinal application
across various diseases.

Keywords: Antioxidant activity, Cedrus atlantica, Organic fractions, Phytochemical analysis,

Polyphenols.

Introduction Cedrus atlantica (Endl.) G. Manetti ex, commonly referred to as Atlas

cedar, represents a significant species of forest tree prevalent in North
Since antiquity, humans have employed medicinal and Africa. It belongs to the Pinaceae family, the subfamily of Abieteae, and
aromatic plants to cure a wide range of ailments.1 Over the years, the Cedrus Genre,7 and one of the most commercially and
medicinal and aromatic plants have seen significant development due environmentally significant plants in the Mediterranean highlands of
to increasing demand from the world market. Morocco.
Morocco, with its strategic geographical location, and a unique Saharan Within traditional medicine, C. atlantica is employed in the treatment
and Mediterranean climates favours a significant diversity of flora of numerous diseases, including parasitic infections in animals, 8
spanning various botanical families,2 a flora that contains a multitude cancer,9 cellulite,10 stress, anxiety, and tension.11 Studies have
of rare, endemic, or notably unique species.3 demonstrated its anti-inflammatory properties and efficacy in
Many aromatic and medicinal plants found in the different Moroccan alleviating symptoms of hay fever.12 Additionally, it possesses
regions have therapeutic values that have been supported by scientific mucolytic properties and is employed in addressing ailments like
evidence.4,5 According to WHO guidelines from 2003 (World Health catarrh, chronic bronchitis, and cough.12,13 It is also effective in the
Organization), traditional medical treatments are utilized by 80% of the treatment of cystitis and urinary tract infection,11,13 and as hair and skin
global population to fulfill their primary healthcare requirements. The care product for oily skin, dandruff, seborrhoea of the scalp, and
pharmacopeia incorporates a vast array of over 20,000 plants, while acne.11,13 Phytochemical investigation showed that C. atlantica contains
natural sources contribute to more than half of the pharmaceutical several classes of bioactive compounds, including sesquiterpene
products commercially available.6 hydrocarbons, monoterpene hydrocarbons, and oxygenated
*Corresponding author. E mail: [email protected] monoterpenes.14–36
Tel: +212620054405 Studies have explored the diverse biological activities of C. atlantica,
encompassing its antibacterial properties,32,35,37 anticancer,35,38–40
Citation: Ameggouz M, Drioua S, El-Guourrami O, Azalmad H, Ouajdi M, analgesic,19,27 antifungal,21,41,42 antiparasitic,22,25,30,31,43 as well as
Zahidi A, Doukkali A, Satani B, Benzeid H. Phytochemical Analysis and antioxidant activities.33,35,44–47
Evaluation of the Antioxidant Activity of Cedrus atlantica (Endl.) G. This study aims to evaluate the antioxidant properties and conduct an
Manetti ex Carrière Stem Extracts. Trop J Nat Prod Res. 2024; 8(3):6741- exhaustive phytochemical analysis of extracts derived from C. atlantica
6750. https://doi.org/10.26538/tjnpr/v8i3.40
originating in Morocco. In spite of its pharmacological capabilities, this
species remains largely underexploited, particularly in terms of organic
Official Journal of Natural Product Research Group, Faculty of Pharmacy,
University of Benin, Benin City, Nigeria extracts. Current evaluations of antioxidant activity are limited to three

6741
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

standard tests, which are insufficient to fully demonstrate the plant's Determination of Total Flavonol Content (TFC-2)
potential. This work seeks to address this gap and identify new chemical The quantification of the total amount of flavonols followed the
compounds with remarkable antioxidant properties. procedure outlined in the study by Yermakov et al. (1987).54 In brief, a
mixture of 6 mL of 5% sodium acetate solution and 2 mL of 2%
aluminum chloride (AlCl3) solution was prepared and then combined
Materials and Methods with 2 mL of each extract. Following a 2-hour and 30-minute incubation
Chemicals period at room temperature, the absorbance of the mixture was assessed
The chemicals used throughout the present study are of high analytical at 440 nm. The standard used to create the calibration curve was
quality. Cyclohexane, ethyl acetate, ethanol, iron chloride, sodium quercetin (15–250 µg/mL). The total flavonols content was quantified
hydroxide, Dragendorff’s reagent, Mayer’s reagent, hydrochloric acid, and stated as milligrams of QE (quercetin equivalents) per gram of dry
sulfuric acid, chloroform, and methanol were procured from Sigma weight (d.w.) of the extract.
Aldrich.
Determination of Total Tannin Content (TTC)
Plant material The quantification of total tannin content in each extract was conducted
Cedar stem were harvested from Atlas Mountain in the Ifrane region of using the methodology detailed by Julkunen-Tiitto (1985).55 A solution
Morocco (geographical coordinates: 33° 29' 54.2'' N5° 08' 06.2'' W) in comprising 750 µL of concentrated hydrochloric acid (37%) and 1.5 mL
October, 2021. of a 4% methanol solution of vanillin was prepared, into which 50 µL
The botanical sample has been validated by the Scientific Center in of each extract was introduced. The mixture was incubated in darkness
Rabat, where it was cataloged in the institute's herbarium, and assigned at room temperature for a duration of 20 minutes. Following incubation,
the voucher specimen number RAB114017. The stems were air-dried the absorbance of the reaction mixture against a methanol blank was
under shade conditions at ambient temperature before being finely determined at 500 nm wavelength. A standard calibration curve was
ground into a powder with a particle size of less than 25 µm. established using catechin (100 - 600 µg/mL). The concentration of
total tannins was expressed as mg of Catechin equivalent/ gram of
Extraction of plant material extract.
The plant material, in powdered form weighing 50 g, was introduced
into a cotton cellulose cartridge and extracted successively with 400 mL Determination of antioxidant capacity
of various solvents (cyclohexane, ethyl acetate, and ethanol in that Determination of DPPH radical scavenging activity
order) employing the Soxhlet apparatus. The extraction process lasted The antioxidant activity of C. atlantica extracts was evaluated using the
6 hours for hexane and 8 hours for ethyl acetate and ethanol, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, as
emergence of colorless solvents in the siphon tube signaled the described by Şahin et al. (2004).56 In a concise procedure, each extract
conclusion of the extraction procedure. Following this, the residual at varying concentrations (50 μL) was introduced into 2 mL of 60 μM
plant material underwent maceration with 1000 mL of distilled water DPPH solution dissolved in methanol. The solution was allowed to sit
for 8 hours under dark conditions at room temperature. The crude undisturbed for 20 minutes at room temperature in the absence of light.
extracts underwent filtration utilizing Whatman filter paper, followed The reaction mixture's absorbance was quantified at 517 nm. The
by concentration through a rotary evaporator (GREATWALL R-1001 measurement of absorbance was also performed for a control solution.
Rotavapor WB-2000 Water bath, China) under reduced pressure, with Quercetin was utilized as the standard antioxidant compound, with
a bath temperature maintained at 35–40°C and rotating at a speed of 120 concentrations ranging from 0.38 to 6.09 mg/mL. The DPPH radical
rpm. After that, the extracts were kept at 4°C until they were needed. 48 scavenging activity percentage was computed followed this formula:
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝐵𝑙𝑎𝑛𝑘 – 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝑇𝑒𝑠𝑡
% 𝐼𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 𝑜𝑓 𝐷𝑃𝑃𝐻 = ×
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝐵𝑙𝑎𝑛𝑘
Phytochemical screening
100 (1)
An initial phytochemical screening was carried out on the extracts in
The IC50 was determined from the regression equation obtained by
order to determine secondary metabolites within the plant. The
plotting scavenging activity percentage against concentration.57
qualitative phytochemical analysis was based on color changes and/or
precipitation following previously reported procedures. 49–51
Determination of ABTS radical scavenging activity
ABTS radical scavenging activity was evaluated according to the
Quantitative phytochemical analysis
method outlined by Pukalskas et al. (2002).58 The ABTS radical was
Determination of Total Phenolic Content (TPC)
generated by mixing ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-
The Folin-Ciocalteu technique, as defined by Poh Hwa et al. (2011),52
sulfonic acid) solution at 7 mM with an equal volume of potassium
was employed to determine the total amount of phenolic compounds. In
persulfate solution at 70 mM. The combination was allowed to incubate
summary, each extract (200 µL) was blended with 1000 µL of 10%
at room temperature in darkness for a duration of 16 hours. Before usage
Folin-Ciocalteu reagent. 800 µL of 7.5% Sodium Carbonate (Na 2CO3)
in the experiment, the ABTS cation radical solution was diluted with
was added to neutralize the reaction. Following a 30-minute incubation
methanol to achieve an absorbance reading of approximately 0.700 at
period in darkness, absorbance readings at 765 nm against a blank were
734 nm. Under the experimental conditions, each extract at different
obtained using a UV-6300PC spectrophotometer. A calibration curve
concentrations was mixed with 2 mL of the ABTS cation radical
was constructed using freshly prepared solutions of gallic acid (7 - 125
solution.59 The absorbance was recorded at 734 nm, and the percentage
µg/mL). The regression equation derived from the garlic acid
radical scavenging activity was determined utilizing the formula below.
calibration curve was used to compute the total phenolic content, and
the result was represented as mg of garlic acid equivalents (GAE)/g of 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝐵𝑙𝑎𝑛𝑘 – 𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝑇𝑒𝑠𝑡
extract dry weight (d.w.). % 𝐼𝑛ℎ𝑖𝑏𝑖𝑡𝑖𝑜𝑛 𝑜𝑓𝐴𝐵𝑇𝑆 = ×
𝐴𝑏𝑠𝑜𝑟𝑏𝑎𝑛𝑐𝑒 𝑜𝑓 𝐵𝑙𝑎𝑛𝑘
100 (2)
Determination of Total Flavonoid Content (TFC-1)
The total flavonoid content was determined using the aluminum Ferric reducing antioxidant power (FRAP) assay
chloride colorimetric method as previuosly described by Ordoñez et al. The spectrophotometric determination of ferric reducing antioxidant
(2006).53 A 0.5 mL solution of each extract was mixed with 0.5 mL of power (FRAP) activity in the extracts was conducted according to the
2% Aluminum Chloride (AlCl3) solution. After incubating for one hour procedure elucidated by Oyaizu (1986).60 A quantity of 0.2 mL of the
at ambient temperature, at 420 nm the absorbance of the mixture was extract or standard at varying concentrations was mixed with 2.5 mL of
assessed, relative to a methanol blank. A calibration curve was 0.2 M sodium phosphate buffer with pH = 6.6 and 2.5 mL of 1% (w/v)
constructed using quercetin (10 - 60 µg/mL). The total flavonoid potassium ferricyanide [K3Fe(CN)6]. Following incubation at 50°C for
content was quantified and stated as milligrams of QE (quercetin 20 minutes, 2.5 mL of 10% (w/v) trichloroacetic acid was added to the
equivalents) per gram of dry weight (d.w.) of the extract. mixture. Then, 2.5 mL of distilled water and 0.5 mL of 0.1% (w/v) ferric
chloride (FeCl3) were added to 2.5 mL of the resulting mixture. The

6742
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

intensity of the resulting blue-green coloration was tracked by assessing The selection of solvent in the extraction process is responsible for the
the absorbance at 700 nm. Catechin (0.65 - 21.39 µg/mL) was employed variation in yields of stem extracts. The polarity of the solvents utilized
as a reference compound. in the extraction process influences the effectiveness of the extraction.
The ethanol extract (F3) exhibited the highest yield, registering at
Determination of Total Antioxidant Capacity (TAC) 14.25%, followed by the aqueous extract (F4) with a yield of 11.78%,
The overall antioxidant capacity of C. atlantica extracts was evaluated the Ethyl acetate extract (F2, yield = 6.85%), and the cyclohexane
using the method for phosphomolybdenum complex formation extract (F1, yield = 2.44 %). The variation in the yield is most likely
developed by Prieto et al. (1999).61 To each extract, 0.2 mL was added due to the differences in the extraction procedures, extraction time, and
to 2 mL of the reagent solution consisting of 0.6 M sulfuric acid, 4 mM solvent polarity. According to recent studies, the choice of solvent
ammonium molybdate, and 28 mM sodium phosphate. Following affects phenolic content, extraction yield, and consequently, the
boiling for 90 minutes at 95°C, the measurement of absorbance for the biological characteristics of plant-based extracts.63–65
resulting blend was conducted at 695 nm. The standard substance
utilized was ascorbic acid (15 - 250 µg/mL). The overall antioxidant Phytochemical constituents of C. atlantica stem
capacity was denoted in (mg AAE/g extract) micrograms of (AAE) Phytochemical screenings are qualitative tests that use staining and
ascorbic acid equivalent per gram of extract.62 precipitation with specialized reagents to identify the distinct families
of secondary metabolites present in plants. These examinations are
GC-MS analysis linked to the development of precipitates, hues, and their magnitudes,
The analysis of C. atlantica extracts via GC-MS took place at MAScIR: which vary in accordance with the concentration of the analyzed
Moroccan Foundation Institute for Advanced Science, Innovation, and compound. Table 2 reports the phytochemical constituents within in the
Research using an Agilent 7890A Series gas chromatography coupled powdered stem of C. atlantica and their extracts.
with mass spectrometry. In the GC-MS system, a 123-BD11 column Secondary metabolites are extensively investigated, particularly in the
(15 m × 320 μm × 0.1 μm), and a multimode injector were employed. case of plant polyphenols, which are widely used in therapies as
The 4 μL of soluble extract was injected into the column with a flow antiparasitic, anti-inflammatory, and antioxidants agents. In the present
rate set at 2 mL/min, employing split mode 1/4 and utilizing helium as study, flavonoids, tannins, and coumarins were among the polyphenolic
the carrier gas. The composition of the extracts was analyzed by compounds detected in C. atlantica stem.
determining the ratio of all compounds present in the sample, The occurrence of flavonoids, tannins, anthocyanins, coumarins,
employing the full scanning mode spanning from 30 to 1000 m/z, using saponins, and quinones was revealed by the cyclohexane and ethyl
an electron impact ionization and a gain factor of 5. The ionization acetate extracts. The ethanol-based extract unveiled the existence of all
source and quadrupole temperatures were configured to 230°C and secondary metabolites tested except quinones and alkaloids, While the
150°C, correspondingly, as for the oven, the temperature program aqueous extract only displayed sterols, flavonoids, tannins, coumarins,
commenced at 30°C and concluded at 360°C. Compounds were and saponins.
identified utilizing the MS NIST 2017 library. An examination of the crude powdered sample demonstrates the
existence of flavonoids, anthocyanins, tannins, coumarins, terpenoids,
Analysis of statistical data quinones, anthraquinones, sterols, and saponins. The discovery of
Statistical analysis was conducted with GraphPad Prism v8. The different secondary metabolite classes in plants enables the anticipation
information was depicted as the average ± standard deviation derived of their pharmacological effects.
from three repeated assessments. A one-way analysis of variance
(ANOVA) has been conducted on the data. The Tukey's post hoc test Table 1: Extraction conditions and yields of the extracts of C.
was employed to analyze the discrepancies among the mean values. atlantica stem
Significant statistical disparities were observed at a p-value threshold
of ≤ 0.05. Extract Code Extraction Method Solvent Yield (%)
F1 Soxhlet method Cyclohexane 2.44%
Results and Discussion F2 Soxhlet method Ethyl Acetate 6.85%
Extraction yield F3 Soxhlet method Ethanol 14.25%
Assessing yields provides an advantage in gauging the optimal amount
of the specific medicinal substance to be harvested from the F4 Maceration Water 11.78%
environment, ensuring the responsible management of natural F1: Cyclohexane extract; F2: Ethyl acetate f extract; F3: Ethanol
reservoirs. The extraction percentages of C. atlantica extracts are extract; F4: Aqueous extract
presented in Table 1.

Table 2: Phytochemical constituents of the extracts of C. atlantica stem

Extracts/Compounds Crude powdered sample Cyclohexane extract Ethyl acetate extract Ethanol extract Aqueous extract
Flavonoids + + + + +
Tannins + + + + +
Anthocyanins + - - + -
Coumarins + + + + +
Quinones + + + - -
Terpenoids + - - + -
Alkaloids - - - - -
Anthraquinones + - - + -
Saponins + + + + +
Sterols + - - + +
+ indicates presence of the compound; - indicates absence of the compound

6743
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

Quantitative phytochemical constituents of C. atlantica stem control, quercetin (IC50 = 5.49 ± 0.02 μg/mL). The ABTS assay showed
The spectrophotometric method was employed to quantify the that C. atlantica extracts had strong antioxidant activity, IC50 = 21.24 ±
phytochemical constituents present in the extracts of C. atlantica stem, 0.002 μg/mL for ethanol, 53.90 ± 0.02 μg/mL for ethyl acetate, and
with the objective of evaluating the overall levels of polyphenol, tannin 57.43 ± 0.006 μg/mL for aqueous extract. By contrast, the extract of
contents, flavonoid, and flavonol. The decision to assess these cyclohexane demonstrated significantly less activity (IC50 = 4571.33 ±
phytoconstituents was prompted by their acknowledged role in 9.57 μg/mL). In statistical terms, it's noteworthy that all the extracts
enhancing the antioxidant potential of therapeutic herbs. Table 3 displayed significantly lower antioxidant activity than ascorbic acid, the
presents the findings. IC50 is 2.52 ± 0.02 μg/mL.
For assessing the reduction capability of the extracts (F1, F2, F3, and
The assessment of total polyphenol content revealed significantly F4) of C. atlantica stem, two tests were carried out; (i) the ferric ion
higher levels in the ethanol (F3) and ethyl acetate (F2) extracts, with reduction (FRAP) assay, and the molybdate reduction (TAC) assay. The
respective amounts of 237.23 ± 1.61, and 132.02 ± 0.60 mg GAE/g results obtained from the FRAP experiment demonstrate that both
extract, unlike the cyclohexane (F1) and aqueous (F4) extracts, which ethanol and ethyl acetate extracts exhibit a considerably higher ability
showed lower levels of 12.94 ± 0.17 mg, and 8.96 ± 0.14 mg GAE/g to reduce ferrous ion (Fe+3) compared to the aqueous and cyclohexane
extract, respectively, with statistical significance (p < 0.05). There was extracts. Thus, the sequence of declining FRAP activity is as follows:
noticeably more tannin in the ethanol extract (189.40 ± 0.76 mg CE/g Ethanol extract (EC50 = 94.36 ± 2.51 μg/mL) > Ethyl acetate extract
extract) compared to the ethyl acetate, cyclohexane, and aqueous (EC50 = 138.18 ± 0.43 μg/mL) > aqueous extract (EC50 = 432.36 ± 2.55
extracts (66.32 ± 2.49, 92.76 ± 1.31, and 25.36 ± 0.61 mg CE/g extract, μg/mL) > Cyclohexanoic extract (EC50 = 1018.89 ± 2.33 μg/mL). It is
respectively), with statistical significance (p < 0.05). Highlighting the notable that the positive control, catechin (EC50 = 13.90 ± 0.03 μg/mL),
fact that ethanol exhibits the best extraction efficiency is crucial. Studies exhibited significantly greater efficacy compared to each of the four
in the literature have demonstrated that ethanol has an excellent extracts (p < 0.05).
capacity to extract phenolic chemicals from various plant sources. 64,66,67 These findings indicate that the ethanol and ethyl acetate extracts
The ethyl acetate and cyclohexane extracts displayed notable levels of demonstrate notable reducing power and contains molecules with a
total flavonoids (81.53 ± 1.13, and 80.62 ± 0.57 mg QE/g extract, stronger electron-donating potential. The polyphenols contained in the
respectively) in comparison to the remaining two extracts, with ethanol and ethyl acetate extracts are probably responsible for the
statistical significance (p < 0.05). Additionally, it was uncovered that antioxidant properties observed in these extracts, potentially due to their
the extract of cyclohexane possessed the greatest level of flavonols capability to attach to metal ions and scavenge free radicals.
(46.49 ± 0.06 mg QE/g extract), followed by the ethyl acetate extract The assessment of total antioxidant capacity followed the technique
(33.4 ± 0.16 mg QE/g extract), then the ethanol extract (6.17 ± 0.16 mg using ammonium molybdate. The Table 4 displays the finding,
QE/g extract), conversely, the smallest quantity of flavonols was indicating that the ethanol extract emerges as the most potent,
present in the aqueous extract. (2.40 ± 0.01 mg QE/g extract). showcasing a total antioxidant capacity (TAC) value of 780.97 ± 4.86
An evident and noteworthy correlation (r = 0.923) was found between mg AAE/g. Following this, the ethyl acetate and aqueous extracts
the overall phenol and tannin levels, indicating that tannins were the exhibit TAC values of 434.95 ± 18.88 mg AAE/g and 429.22 ± 4.21 mg
predominant phenolic constituents extracted from the stem of C.
AAE/g, respectively. The extract with the least total antioxidant
atlantica. A hight correlation (r = 0.965) was also observed for the
capacity was the cyclohexane one, registering a TAC value of 366.65 ±
concentrations of total flavonols and total flavonoids.
6.36 mg AAE/g, notably lower than that of the other extracts (p < 0.05).
The pronounced antioxidative efficacy demonstrated by the ethanol,
Antioxidant activity of C. atlantica stem extracts
ethyl acetate, and aqueous extracts from C. atlantica stem across
Currently, there is no standardized technique available for assessing the
multiple assays DPPH, ABTS, FRAP, and TAC, this could be explained
antioxidant efficacy of plant extracts. Consequently, it is advisable to
by the plant's phenolic compound content, particularly flavonoids.
employ multiple methodologies to offer comprehensive insights. 68 In
Phenolic compounds are known as antioxidant substances having the
the current study, four techniques were utilized to assess the antioxidant
ability to scavenge radical species and reactive forms of oxygen. The
activity in vitro of various C. atlantica extracts: DPPH and ABTS,
outcomes obtained from the assessment of total phenolic content, which
which measure radical scavenging capacity, along with the FRAP test
revealed significant amounts of phenolic substances in the ethyl acetate
for ferric reducing antioxidant power and the molybdate test (TAC).
and ethanol extracts of C. atlantica stems, validate the aforementioned
Table 4 displays the outcomes of the antioxidant performance
claim.
assessment of C. atlantica stem extracts alongside standards, conducted
Belkacem et al. (2021)35 studied the antioxidative potential of extracts
through the DPPH, ABTS, FRAP, and TAC assays.
(acetone, ethanol, and methanol) as well as fractions (chloroform, ethyl
The findings regarding the antioxidant properties of DPPH indicate that
acetate, n-butanol, and aqueous) produced from C. atlantica's stem
the ethanol extract exhibits superior ability in neutralizing DPPH
using the DPPH, FRAP, and ABTS assays. According to their findings,
radicals than the other extracts. The IC50 values were 19.40 ± 0.01, the n-butanol and ethyl acetate fractions demonstrated the strongest
45.60 ± 0.01, and 86.17 ± 0.25 μg/mL for the ethanol extract, ethyl antioxidant effectiveness, trailed by the acetone, ethanol, and methanol
acetate extract, and aqueous extract, respectively. Nonetheless, it’s extracts, which displayed moderate activity.
important to note that the extracts exhibited significantly lower DPPH
radical scavenging activity (p < 0.05) comparing with the positive

Table 3: Polyphenol, flavonoid, flavonol and condensed tannin contents of extracts of C. atlantica stem
Plant Extracts TPC TFC-1 TFC-2 TTC
(mg GAE/g Extract) (mg QE/g Extract) (mg QE/g Extract) (mg CE/g Extract)
F1 12.94 ± 0.17a 80.62 ± 0.57e 46.49 ± 0.06h 92.76 ± 1.31l
F2 132.02 ± 0.60b 81.53 ± 1.13e 33.4 ± 0.16i 66.32 ± 2.49m
F3 237.23 ± 1.61c 16.35 ± 0.15f 6.17 ± 0.16j 189.4 ± 0.76n
F4 8.96 ± 0.14d 5.47 ± 0.06g 2.40 ± 0.01k 25.36 ± 0.61o
Data represent the mean ± standard deviation of three independent experiments. Values in the same column with different superscript letters indicate
significant differences (p-value < 0.05). F1: Cyclohexane extract; F2: Ethyl acetate extract; F3: Ethanol extract; F4: Aqueous extract; TPC: Total Phenolic
Content; TFC-1: Total Flavonoid Content; TFC-2: Total Flavonol Content; TTC: Total Tannins Content.

6744
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

Table 4: Antioxidant activity of extracts of C. atlantica stem and standards

Plant Extracts DPPH ABTS FRAP TAC
IC50 (μg/mL) IC50 (μg/mL) EC50 (μg/mL) mg AAE/g of extract
F1 Nd 4571.33 ± 9.57e 1018.89 ± 2.33g 366.65 ± 6.36l
a f h
F2 45.60 ± 0.01 53.9 ± 0.02 138.18 ± 0.43 434.95 ± 18.88m
F3 19.40 ± 0.01b 21.24 ± 0.002f 94.36 ± 2.51i 780.97 ± 4.86n
F4 86.17 ± 0.25c 57.43 ± 0.006f 432.36 ± 2.55j 429.22 ± 4.21m
Quercetin 5.49 ± 0.02d - - -
Ascorbic acid - 2.52 ± 0.02f - -
k
Catechin - - 13.90 ± 0.03 -
Data represent the mean ± standard deviation of three independent experiments. Values in the same column with different superscript letters indicate
significant differences (p-value < 0.05). F1: Cyclohexane extract; F2: Ethyl acetate extract; F3: Ethanol extract; F4: Aqueous extract; IC50: 50% inhibitory
concentration; EC50: Effective concentration that transforms 50% of Fe3+ into Fe2+, Nd = Not determined.

A B

C D
Figure 1: GC Chromatograms of C. atlantica extracts. A: Cyclohexane extract, B: Ethyl acetate extract, C: Ethanol extract, D: Aqueous
extract.

6745
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

In contrast, the chloroform and aqueous fractions demonstrated low diphenyl- (3.74%), Ethanol, 2,2'-[1,2-ethanediylbis(oxy)] bis-,
levels of activity. However, the work of Fadel et al. (2016)46 diacetate (3.58%), and Clionasterol (3.21%) - a phytosterols which is a
demonstrated that the ethanolic extract obtained from the aerial portions poriferast-5-ene carrying a beta-hydroxy substituent at position 3.
of C. atlantica exhibited significant antioxidative potential, as assessed Clionasterol stands as a significant metabolite found in both plants and
by DPPH, having an 8.92 ± 0.35 μg/mL IC50 value. marine organisms. It constitutes a 3-beta-sterol, belonging to the
phytosterols, and derives from a poriferate hydride, characterized by a
Compounds identified in C. atlantica extract by GC-MS 3-beta-hydroxy-Delta(5)-steroid structure. Clionasterol is a natural
Several components of the C. atlantica extracts were determined via gas product found in Clerodendrum infortunatum, Achillea santolina and
chromatography (GC) in conjunction with mass spectrometry (MS). other organisms.74 The examination of the ethanol extract unveiled the
The chromatograms are depicted in Figure 1, and Table 5 presents the existence of three compounds; Thiophene, tetrahydro-2-methyl-
molecular information of the compounds found in the plant extracts, (7.50%), 9-Octadecenoic acid (Z)-, 2,3- dihydroxypropyl ester (0.16%),
including their molecular weights (MW), chemical names, retention and 1,3-Dioxane, 2-methyl- (0.08%).
times (RT), peak areas, and molecular formulas. GC-MS analysis of C. The GC-MS examination of the plant's aqueous extract detected 23
atlantica plant extracts revealed 20 compounds in the cyclohexane compounds, with Catechol (32.25%), 4-(4-Methyl-
extract, with the major compound being Palustric acid (17.48%), which [1,3,2]dioxaborinan-2-yloxy)-phenol (6.76%), and 2-Pyridinamine,
belongs to the resin acids class of compounds. It has been demonstrated 4,6-dimethyl- (6.57%), as the major compounds. Catechol has proven
that palustric acid possesses potent antibiotic properties. 69 Several to be a pharmacological agent with antimicrobial, astringent, wound
molecules belonging to the resin acids group have known biological healing and insecticidal activities.75 Resorcinol which is known for its
activities such as antibacterial,70 antifungal,71 antitumor, antiviral, nitric antipruritic, antimycotic and antiseptic properties,76 was present in
oxide inhibitory,72 and larvicidal activities.73 minor quantity The results of this study reflect therapeutic promise in
GC-MS analysis of C. atlantica plant extracts revealed 7 compounds in the extracts derived from the stem of C. atlantica, supporting its
the ethyl acetate extact, the most significant ones being acetamide, 2- traditional application in Moroccan herbal medicine.
(5,7-dimethyl-[1,2,4]triazolo[1,5-a]pyrimidin-2-ylsulfanyl)-N,N-

Table 5: Compounds identified in C. atlantica extracts by GC-MS

S/N Library/ID RT (min) Peak Area (%) M.W. Molecular formula
Cyclohexane extract
1 Caryophyllene 8.166 1.52 204.35 C15H24
2 (1R,4aR,4bS,7R,10aR)-1,4a,7- Trimethyl-7-vinyl- 23.334 0.81 286.451 C20H30O
1,2,3,4,4a,4b,5,6,7,9,10,10a-dodecahydro-
phenanthrene-1-carbaldehyde
3 1-Phenanthrenecarboxaldehyde 1,2, 144480 013601- 24.371 1.03 284.435 C20H28O
88-2 99 3,4, 4a,9,10,10a-octahydro-1,4a-dim ethyl-7-
(1-methylethyl)-, [1R-(1.a
lpha.,4a.beta.,10a.alpha.)]-
4 Methyl dehydroabietate- 25.712 0.48 314.461 C21H30O2
5 (1R,4aR,4bS,10aR)-1,4a- Dimethyl-7-(propan-2- 25.802 0.72 286.451 C20H30O
ylidene)- 1,2,3,4,4a,4b,5,6,7,9,10,10a-
dodecahydrophenanthrene-1-carbaldehyde
6 Palustric acid 27.188 17.48 302.5 C20H30O2
7 1-Hydroxy-3,5,8-trimethoxy- xanthen-9-one 27.425 9.38 302.28 C16H14O6
8 Dehydroabietic acid 27.954 10.25 300.4 C20H28O2
9 13-Tetradece-11-yn-1-ol-9-Octadecyne 23.773 5.01 208.34 C14H24O
10 Abietic acid 28.382 7.62 302.5 C20H30O2
11 1-Phenanthrenecarboxylic acid, 7-ethenyl- 29.182 10.39 316.477 C21H32O2
1,2,3,4,4a,4b,5,6,7,9, 10,10a-dodecahydro-1,4a, 7-
trimethyl-, methyl ester, [1R-(1α,4aβ,4bα,7α,10aα)]-
12 2,4,5,7-tetramethoxy-9H-fluorene 29.937 2.71 286.32 C17H18O4
13 2,2,4,4,5,5,7,7-Octamethyl 30.445 2.58 294.686 C10H30
14 Tetracosanal 30.839 1.19 352.6 C24H48O
15 Methyl palustrate 31.188 1.70 316.5 C21H32O2
16 1-Docosene 35.583 1.36 308.6 C22H44
17 Vitamin E 36.834 0.53 430.7 C29H50O2
18 Triallylmethylsilane 37.048 0.91 166.33 C10H18Si
19 Tetrapentacontane, 1,54-dibromo- 38.107 0.70 917.2 C54H108Br2
20 Clionasterol 38.321 1.71 414.7 C29H50O
Ethyl acetate Extract
1 Benzoic acid 5.733 1.59 122.12 C7H6O2
2 Ethanol, 2,2'-[1,2-ethanediylbis(oxy)] bis-, diacetate 21.261 3.58 234.246 C10H18O6
3 2-Dodecen-1-yl(-)succinic anhydrid 23.717 1.74 266.38 C16H26O3
4 Dehydroabietic acid 27.504 1.34 300.4 C20H28O2
5 Acetic acid, 11,11-dimethyl-3,4- dihydro-2H-1,4- 28.923 0.85 296.37 C18H20N2O2
methanophenazin-1-ylm ethyl ester

6746
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

6 Acetamide, 2-(5,7-dimethyl-[1,2,4]triazolo[1,5- 30.433 3.74 343.4 C16H17N5O2S

a]pyrimidin-2-ylsulfa nyl)-N,N-diphenyl-
7 Clionasterol 38.299 3.21 414.7 C29H50O
Ethanol Extract
1 1,3-Dioxane, 2-methyl- 1.631 0.08 102.13 C5H10O2
2 Thiophene, tetrahydro-2-methyl- 22.320 7.50 102.20 C5H10S
3 9-Octadecenoic acid (Z)-, 2,3- dihydroxypropyl ester 48.745 0.16 356.539 C21H40O4
Aqueous Extract
1 Propanoic acid, 2-chloro-, ethyl ester 12.366 0.72 136.57 C5H9ClO2
2 4H-Pyran-4-one, 2,3-dihydro-3,5-dihydroxy-6- 12.659 1.78 144.12 C6H8O4
methyl-
3 Benzoic acid 12.862 5.68 122.12 C7H6O2
4 Catechol 13.188 32.25 110.11 C6H6O2
5 Benzofuran, 2,3-dihydro- 13.402 4.60 120.15 C8H8O
6 Benzeneacetic acid 13.797 0.67 136.147 C8H8O2
7 1,2-Benzenediol, 3-methyl- 13.977 0.60 124.13 C7H8O2
8 4-(4-Methyl-[1,3,2]dioxaborinan-2-yloxy)-phenol 14.135 6.76 208.019 C10H13
9 2-(2-Hydroxyethoxy)phenol 14.394 4.06 154.16 C8H10O3
10 2-Methoxy-4-vinylphenol 14.856 3.57 150.17 C9H10O2
11 3(2H)-Pyridazinone, 6-methyl- 14.991 1.39 110.11 C5H6N2O
12 2,3,5-trithiahexane 5-oxide 15.555 1.28 156.3 C3H8OS3
13 Benzoxazole, 2-methyl- 15.701 2.17 133.15 C8H7NO
14 2-Pyrrolidinophenol 16.603 0.89 179.22 C10H13NO2
15 2,4,5-trithiahexane 2,2-dioxide 17.177 0.63 172.28 C3H8O2S3
16 Resorcinol 17.369 0.38 110.11 C6H6O2
17 Acetamide, 2-(adamantan-1-yl)- N-(1-adamantan-1- 21.651 0.26 355.557 C24H37
ylethyl)-
18 Carbonic acid, 2-ethylhexyl heptadecyl ester 21.944 1.57 426.7 C27H54O3
19 4b-Methyl-6,8-dioxa-3-thia- bicyclo (3,2,1)octane 22.045 0.40 146.21 C6H10O2S
20 Decanoic acid, 3-methyl- 23.623 0.51 186.29 C11H22O2
21 Thiophene, tetrahydro-2-methyl- 24.063 0.64 102.20 C5H10S
22 2-Pyridinamine, 4,6-dimethyl- 24.378 6.57 122.16 C7H10N2
23 Thiophene, tetrahydro-2-methyl- 25.257 0.94 102.20 C5H10S

Conclusion References
In the present study, our focus lies in investigating the phytochemical 1. Ait Sidi Brahim M, Markouk M, Larhsini M. Moroccan
analyses and evaluating the antioxidant effectiveness of various stem Medicinal Plants as Antiinfective and Antioxidant Agents. In
extracts sourced from C. atlantica. The findings reveal that C. atlantica New Look to Phytomedicine: Advancements in Herbal
holds significant pharmacological potential, highlighting its abundance Products as Novel Drug Leads. 2019; p. 91‑142.
in phenolic compounds. The ethanol and ethyl acetate extracts stand https://doi.org/10.1016/B978-0-12-814619-4.00005-7.
out, having the most potent antioxidant capacity, while the cyclohexane 2. Zaher A, Boufellous M, Jaber H, El Hartiti H, Barrahi M,
extract also demonstrated notable activity. These discoveries pave the Ouhssine M, Bourkhiss B. Ethnobotanical Study of
way for new research aimed at exploring the various biological Medicinal Plants Used in the Province of Sidi Slimane
properties of this plant and the identification of its metabolites. (Morocco). JBM. 2018; 06(09):25‑35.
https://doi.org/10.4236/jbm.2018.69003.
3. Najem M, Daoudi A, Bouiamrine EH, Ibijbijen J, Nassiri L.
Conflict of Interest Biodiversity of poisonous medicinal plants solicited in the
Authors declare no conflict of interest traditional phytotherapy of the central Middle Atlas -
Morocco-. Ethnobot. Res. Appl. 2019; 18:1‑22.
http://dx.doi.org/10.32859/era.18.19.1-22.
Authors’ Declaration 4. Salhi N, El-guourrami O, Rouas L, Moussaid S, Moutawalli
The authors hereby declare that the work presented in this article is A, Benkhouili F, Ameggouz M, Alshahrani M, Awadh A,
original and that any liability for claims relating to the content of this Bouyahya A, my el abbes F, Cherrah Y. Evaluation of the
article will be borne by them. Wound Healing Potential of Cynara humilis Extracts in the
Treatment of Skin Burns. eCAM. 2023; 2023:1‑12.
https://doi.org/10.1155/2023/5855948.
Acknowledgments 5. Mahraz A M, Elhachmia C, Zakia R, Taleb M. Medicinal
Plants of Moulay Yaâcoub Province in Morocco: An
The authors thank Professor Mohammed Sghir Taleb at the Scientific
Ethnobotanical and Biodiversity Survey. TJNPR. 2023; 7(8).
Institute of Rabat for the botanical identification of the plant material.
http://www.doi.org/10.26538/tjnpr/v7i8.3.
6. Hamilton AC. Medicinal plants, conservation and
livelihoods. Biodivers. Conserv. 2004; 13(8):1477‑517.
7. Serbouti S, Abbas Y, Soussi M, Alaoui I, Squalli W, Achiban
H. Morphological Responses of Cedrus atlantica, Pinus

6747
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

halepensis, and Tetraclinis articulata in Different Silva A S. In vitro effect of seven essential oils on the
Pedoclimatic Conditions. TJNPR. 2022; 6(12): 1919–1924. reproduction of the cattle tick Rhipicephalus microplus. J
http://www.doi.org/10.26538/tjnpr/v6i12.3. Adv Res. 2016;7(6):1029‑34.
8. Bellakhdar J. Jamal Bellakhdar - La pharmacopée marocaine https://doi.org/10.1016/j.jare.2016.05.003.
traditionnelle - Jamal Bellakhdar. Éditions Le Fennec 23. Benouaklil F, Hamaidi-Chergui F, Hamaidi Ms, Saidi F.
[Online]. 1997 [cited 2022 Jan 31]. Available from: Chemical Composition and Antimicrobial Properties of
https://lefennec.com/livre/la-pharmacopee-marocaine- Algerian Cedrus Atlantica M. Essential Oils | agrobiologia
traditionnelle-jamal-bellakhdar/. [Online]. 2017 [cited 2022 Feb 1]. Available from:
9. Bouayyadi L, El Hafian M, Zidane L. Étude floristique et https://agrobiologia.net/online/chemical-composition-and-
ethnobotanique de la flore médicinale dans la région du antimicrobial-properties-of-algerian-cedrus-atlanticam-
Gharb, Maroc. J App Bioscience. 2015; 93(1):8770. essential-oils/.
https://doi.org/10.4314/jab.v93i1.10. 24. El-miziani I, Sara H, Essahli M, Lhaloui S, Lamiri A. Lead
10. Schnaubelt K. MEDICAL AROMATHERAPY: Healing corrosion inhibition by cedrus atlantica as a green inhibitor
with Essential Oils. Berkeley, CA: Frog [Online]. 1999 [cited in 0.1M Na2CO3 solution. Int. J. Adv. Chem. 2017; 5:1.
2022 Feb 1]. Available from: https://doi.org/10.14419/ijac.v5i1.7115.
https://www.biblio.com/book/medical-aromatherapy- 25. ez zoubi Y, el-akhal F, Farah A, Taghzouti K, El Ouali
healing-essential-oils-schnaubelt/d/1276415526. Lalami A. Chemical composition and larvicidal activity of
11. Lawless J. The encyclopeadia of essential oils: a complete Moroccan Atlas Cedar (Cedrus atlantica Manetti) against
guide to the use of aromatics in aromatherapy, herbalism, Culex pipiens (Diptera: Culicidae). J. Appl. Pharm. Sci.
health & well-being. Shaftsbury, Dorset: Element; 1992. 2017; 7:30‑4. https://doi.org/10.7324/JAPS.2017.70704.
12. Lawrence, B. M. Progress in Essential Oils. Perfumer & 26. Uehara A, Tommis B, Belhassen E, Satrani B, Ghanmi M,
Flavorist [Online]. 2012 [cited 2022 Feb 1]. Available from: Baldovini N. Odor-active constituents of Cedrus atlantica
https://www.perfumerflavorist.com/fragrance/ingredients/ar wood essential oil. Phytochem. 2017; 144:208‑15.
ticle/21859574/progress-in-essential-oils. https://doi.org/10.1016/j.phytochem.2017.09.017.
13. FischerRizzi S. The Complete Aromatherapy Handbook: 27. Emer AA, Donatello NN, Batisti AP, Oliveira Belmonte LA,
Essential Oils for Radiant Health par Susanne Fischer-Rizzi: Santos ARS, Martins DF. The role of the endocannabinoid
Used: Acceptable Paperback (1990) | Lady Lisa’s Bookshop system in the antihyperalgesic effect of Cedrus atlantica
[Online]. 1990 [cited 2022 Feb 1]. Available from: essential oil inhalation in a mouse model of postoperative
https://www.abebooks.fr/9780806982229/Complete- pain. J Ethnopharmacol. 2018; 210:477‑84.
Aromatherapy-Handbook-Essential-Oils-0806982225/plp. https://doi.org/10.1016/j.jep.2017.09.011.
14. Paun G, Zrira S, Boutakiout A, Ungureanu O, Simion D, 28. Belloula N, Dridi S, Khattaf A. Investigation Into The
Chelaru C, et al. CHEMICAL COMPOSITION, Antibacterial Activity And Chemical Composition Of
ANTIOXIDANT AND ANTIBACTERIAL ACTIVITY OF ‘Cedrus Atlantica’ Essential Oil. IND DRU. 2018;
ESSENTIAL OILS FROM MOROCCAN AROMATIC 55(06):51‑5. https://doi.org/10.53879/id.55.06.11292.
HERBS. Rev. Roum. Chim. 2013;9. 29. Uwineza MS, El Yousfi B, Lamiri A. Antifungal activities of
15. Dakir M, El Hanbali F, Mellouki F, Mar Herrador M, Barrero essential oils of Mentha pulegium, Eugenia aromatica and
AF, Benharref A, et al. Chemical and antibacterial studies of Cedrus atlantica on Fusarium culmorum and Bipolaris
essential oils of scales and seeds of Cedrus atlantica Endl. J sorokiniana in vitro. RMPP. 2018; 12: 19-32.
Nat Prod Plant Resour. 2014; 4:15. 30. Ainane A, Benhima R, Khammour F, el Kouali M, Talbi M,
16. Murbach Teles Andrade BF, Nunes Barbosa L, da Silva Abba EH, Cherroud S, Tarik A. Chemical composition and
Probst I, Fernandes Júnior A. Antimicrobial activity of insecticidal activity of five essential oils: Cedrus atlantica,
essential oils. J. Essent. Oil Res. 2014; 26(1):34‑40. Citrus limonum, Rosmarinus officinalis, Syzygium
https://doi.org/10.1080/10412905.2013.860409. aromaticum and Eucalyptus globules. Mater. Today: Proc.
17. Rhafouri R, Satrani B, Zair T, Ghanmi M, Bou-Idra M, 2019; P 474-485.
Omari ME, Bentayeb A. Chemical Composition Of The https://doi.org/10.1016/j.matpr.2019.04.004.
Cedrus Atlantica (Endl.) Manetti Ex Carrière Seeds Essential 31. Alves TJS, Murcia A, Wanumen AC, Wanderley-Teixeira V,
Oil In Function Of Their Germination Stages. IJRAMR. Teixeira ÁAC, Ortiz A, Medina P. Composition and Toxicity
2015;6. of a Mixture of Essential Oils Against Mediterranean Fruit
18. Nam AM, Bighelli A, Ghanmi M, Satrani B, Casanova J, Fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae).
Tomi F. Deodarone Isomers in Cedrus atlantica Essential J Econ Entomol. 2019; 112(1):164‑172.
Oils and Tar Oils. NPC. 2015;10(11). https://doi.org/10.1093/jee/toy275.
https://doi.org/10.1177/1934578X1501001123. 32. Oukhrib A, Zaki M, Ait El Had M, Karroumi J, Bouamama
19. Martins DF, Emer AA, Batisti AP, Donatello N, Carlesso H, Benharref A, Urrutigoïty M. Synthesis of cyclopropane
MG, Mazzardo-Martins L, Venzke D, Micke G A, Pizzolatti ring derivatives from natural β- himachalene and evaluation
M G, Piovezan A P, dos Santos A R S. Inhalation of Cedrus of their antimicrobial activity by bioautography. RHAZES
atlantica essential oil alleviates pain behavior through GAC. 2019; 6:61‑70.
activation of descending pain modulation pathways in a 33. Skanderi I, Chouitah O. Chemical Characterization and
mouse model of postoperative pain. J. Ethnopharmacol. Antioxidant Activity of Cedrus atlantica Manetti Tar (Atlas
2015; 175:30‑8. https://doi.org/10.1016/j.jep.2015.08.048. Cedar Tar). FUJC. 2020; 8(2):244-255.
20. Zrira S, Ghanmi M. Chemical Composition and Antibacterial https://doi.org/10.17721/fujcV8I2P244-255.
Activity of the Essential of Cedrus atlantica (Cedarwood 34. Bennouna F, Lachkar M, EL ABED S, Saad I. Cedrus
oil). J. Essent. Oil-Bear. Plants. 2016; 19:1267‑72. atlantica essential oil: Antimicrobial activity and effect on
https://doi.org/10.1080/0972060X.2015.1137499. the physicochemical properties of cedar wood Surface.
21. Fidah A, Salhi N, Mohamed R, Kabouchi B, Ziani M, Moroccan J. Biol. 2020; 16:35‑45.
Aberchane M, Famiri A. Natural durability of Cedrus 35. Belkacem N, Khettal B, Hudaib M, Bustanji Y, Abu-Irmaileh
atlantica wood related to the bioactivity of its essential oil B, Amrine CSM. Antioxidant, antibacterial, and cytotoxic
against wood decaying fungi. MADERAS-CIENC activities of Cedrus atlantica organic extracts and essential
TECNOL. 2016;18. http://dx.doi.org/10.4067/S0718- oil. Eur. J. Integr. Med. 2021; 42:101292.
221X2016005000049. https://doi.org/10.1016/j.eujim.2021.101292.
22. Pazinato R, Volpato A, Baldissera MD, Santos RCV, Baretta 36. Jaouadi I, Cherrad S, Bouyahya A, Koursaoui L, Satrani B,
D, Vaucher RA, Giongo J L, Boligon A A, Stefani L M, Da Ghanmi M, Chaouch A. Chemical variability and antioxidant

6748
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

activity of Cedrus atlantica Manetti essential oils isolated 51. Gul R, Jan SU, Faridullah S, Sherani S, Jahan N. Preliminary
from wood tar and sawdust. Arab. J. Chem. Phytochemical Screening, Quantitative Analysis of
2021;14(12):103441. Alkaloids, and Antioxidant Activity of Crude Plant Extracts
https://doi.org/10.1016/j.arabjc.2021.10344. from Ephedra intermedia Indigenous to Balochistan. Sci.
37. Thielmann J, Muranyi P, Kazman P. Screening essential oils World J. 2017; 2017:5873648.
for their antimicrobial activities against the foodborne https://doi.org/10.1155/2017/5873648.
pathogenic bacteria Escherichia coli and Staphylococcus 52. Poh Hwa T, Cheah YK, J. IB, Radu S. Bioprotective
aureus. Heliyon. 2019; 5(6):e01860. properties of three Malaysia Phyllanthus species: an
https://doi.org/10.1016/j.heliyon.2019.e01860. investigation of the antioxidant and antimicrobial activities.
38. Saab AM, Lampronti I, Borgatti M, Finotti A, Harb F, Safi Int. Food Res. J. 2011; 18(3):887‑93.
S, Gambari R. In vitro evaluation of the anti-proliferative 53. Ordoñez AAL, Gomez JD, Vattuone MA, lsla MI.
activities of the wood essential oils of three Cedrus species Antioxidant activities of Sechium edule (Jacq.) Swartz
against K562 human chronic myelogenous leukaemia cells. extracts. Food Chem. 2006; 97(3):452‑8.
Nat Prod Res. 2012; 26(23):2227‑31. https://doi.org/10.1016/j.foodchem.2005.05.024.
https://doi.org/10.1080/14786419.2011.643885. 54. Yermakov AI, Arasimov VV, Yarosh NP. Methods of
39. Hung PH, Hsieh MC, Lee SC, Huang XF, Chang KF, Chen biochemical analysis of plants. Agropromizdat, Leningrad.
SY, Lee MS, Tsai NM. Effects of Cedrus atlantica extract 1987; 122–142.
on acute myeloid leukemia cell cycle distribution and 55. Julkunen-Tiitto R. Phenolic constituents in the leaves of
apoptosis. Mol Biol Rep. 2020; 47(11):8935‑8947. northern willows: methods for the analysis of certain
https://doi.org/10.1007/s11033-020-05947-w. phenolics. J Agric Food Chem. 1985; 33(2):213‑7.
40. Chang KF, Chang JT, Huang XF, Huang YC, Li CY, Weng https://doi.org/10.1021/jf00062a013.
JC, Hsiao CY, Hsu HJ, Tsai NM. Cedrus atlantica Extract 56. Şahin F, Güllüce M, Daferera D, Sökmen A, Sökmen M,
Suppress Glioblastoma Growth through Promotion of Polissiou M, Agar G, Özer H. Biological activities of the
Genotoxicity and Apoptosis: In Vitro and In Vivo Studies. Int essential oils and methanol extract of Origanum vulgare ssp.
J Med Sci. 2021;18(11):2417‑30. vulgare in the Eastern Anatolia region of Turkey. Food
https://doi.org/10.7150/ijms.54468. Control. 2004; 15(7):549‑57.
41. Maya BM, Abedini A, Gangloff SC, Kabouche A, Kabouche https://doi.org/10.1016/j.foodcont.2003.08.009.
Z, Voutquenne-Nazabadioko L. A new δ-tocotrienolic acid 57. Drioua S, Azalmad H, El-Guourrami O, Ameggouz M,
derivative and other constituents from the cones of Cedrus Benkhouili FZ, Assouguem A, Kara M, Ullah R, Ali E A,
atlantica and their in vitro antimicrobial activity. Phytochem. Ercisli S, Fidan H, Benzeid H, Doukkali A. Phytochemical
Lett. 2017; 20:252‑8. screening and antioxidant activity of Vitex agnus-castus L.
https://doi.org/10.1016/j.phytol.2017.05.009. Open Chem. 2024; 22(1). https://doi.org/10.1515/chem-
42. Orchard A, Van vuuren S, Viljoen A. Commercial Essential 2023-0190.
Oil Combinations against Topical Fungal Pathogens. Nat. 58. Pukalskas A, van Beek, T.A., Venskutonis, R.P., Linssen,
Prod. Commun. 2019; 14:1934578X1901400. J.P.H., van Veldhuizen, A., de Groot, Æ. Identification of
https://doi.org/10.1177/1934578X1901400139. Radical Scavengers in Sweet Grass (Hierochloe odorata). J.
43. Tennyson S, Samraj DA, Jeyasundar D, Chalieu K. Agric. Food Chem. 2002; 50(10):2914‑9.
Larvicidal efficacy of plant oils against the dengue vector https://doi.org/10.1021/jf011016r.
Aedes aegypti (L.) (Diptera: Culicidae). Middle East J. Sci. 59. Drioua S, El-Guourrami O, Assouguem A, Ameggouz M,
Res. 2013; 13:64‑8. Kara M, Ullah R , Bari A, Zahidi A, Skender A, Benzeid A,
44. Naimi F, Bousta D, Balouiri M, Meskaoui A. Antioxidant Doukkali A.. Phytochemical study, antioxidant activity, and
and free radical-scavenging properties of seeds flavonoids dermoprotective activity of Chenopodium ambrosioides (L.).
extract of Cedrus atlantica Manetti, Linum usitatissimum L. Open Chem. 2024; 22(1). https://doi.org/10.1515/chem-
and Ocimum basilicum L. species. J. Appl. Pharm. Sci. 2015; 2023-0194
5(08):95-99. https://doi.org/10.7324/JAPS.2015.50815. 60. Oyaizu M. Studies on products of browning reaction--
45. El-miziani I, Sara H, Lhaloui S, Essahli M, Lamiri A. Study antioxidative activities of products of browning reaction
of Antioxidant Activity of Essential Oils Extracted from prepared from glucosamine. Eiyogaku zasshi = Japanese
Moroccan Medicinal and Aromatic Plants. European j. med. journal of nutrition [Onligne]. 1986 [cited 2022 Nov 22];
plants. 2015; 10:1‑12. Available from:
https://doi.org/10.9734/EJMP/2015/19955. https://scholar.google.com/scholar_lookup?title=Studies+on
46. Fadel H, Benayache F, Samir B. Antioxidant properties of +products+of+browning+reaction--
four Algerian medicinal and aromatic plants Juniperus antioxidative+activities+of+products+of+browning+reactio
oxycedrus L., Juniperus phoenicea L., Marrubium vulgare n+prepared+from+glucosamine&author=Oyaizu%2C+M.&
L. and Cedrus atlantica (Manetti ex Endl). Sch. Res. J. 2016; publication_year=1986.
8:72‑9. 61. Prieto P, Pineda M, Aguilar M. Spectrophotometric
47. Hofmann T, Visiné Rajczi E, Balázs B, Dániel B, Levente A. quantitation of antioxidant capacity through the formation of
Antioxidant Capacity and Tentative Identification of a phosphomolybdenum complex: specific application to the
Polyphenolic Compounds of Cones of Selected Coniferous determination of vitamin E. Anal Biochem. 1999;
Species. Acta Silv. 2020; 16:79‑94. 269(2):337‑41. https://doi.org/10.1006/abio.1999.4019.
https://doi.org/10.37045/aslh-2020-0006. 62. El-guourrami O, Drioua S, Ameggouz M, Salhi N, Sayah K,
48. Rostagno MA, Prado JM. Natural Product Extraction: Zengin G, Zahidi A, Doukkali A, BENZEID H. Antioxidant
Principles and Applications. RSC; 2013. 1032 p. activity, analgesic activity, and phytochemical analysis of
49. Deyab M, Elkatony T, Ward F. Qualitative and Quantitative Ammi majus (L.) extracts. IJSM. 2023; 10:23‑37.
Analysis of Phytochemical Studies on Brown Seaweed, https://doi.org/10.21448/ijsm.1139246.
Dictyota dichotoma. IJEDR. 2016; 4(2):5. 63. Hadzri HM, Yunus MAC, Zhari S, Rithwan F. The Effects
50. El-Houda N, Daira, Maazi MC, Chefrour A. Contribution à of Solvents and Extraction Methods on the Antioxidant
l’étude phytochimique d’une plante médicinale (Ammoides Activity of P. niruri. Jurnal Teknologi [Online]. 2014 [cited
verticillata Desf. Briq.) de l’Est Algérien. Bull. Soc. R. Sci. 2022 Nov 26];68(5). Available from:
Liege. 2016; 276‑90. https://doi.org/10.25518/0037- https://journals.utm.my/jurnalteknologi/article/view/3030.
9565.6494. 64. Metrouh-Amir H, Duarte CMM, Maiza F. Solvent effect on
total phenolic contents, antioxidant, and antibacterial

6749
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License
 Trop J Nat Prod Res, March 2024; 8(3):6741-6750 ISSN 2616-0684 (Print)
ISSN 2616-0692 (Electronic)

activities of Matricaria pubescens. Ind Crops Prod. 2015; 24(3):199‑205.

67:249‑56. https://doi.org/10.1016/j.indcrop.2015.01.049. https://doi.org/10.3109/02844319009041279.
65. Kamarudin NA, Latip MM and J. Effects of Solvents and 71. Savluchinske Feio S, Gigante B, Roseiro JC, Marcelo-Curto
Extraction Methods on Herbal Plants Phyllanthus niruri, MJ. Antimicrobial activity of diterpene resin acid
Orthosiphon stamineus and Labisia pumila. INDJST. derivatives. J Microbiol Methods. 1999; 35(3):201‑6.
2016;9(21):1‑5. https://doi.org/10.1016/S0167-7012(98)00117-1.
https://doi.org/10.17485/ijst/2016/v9i21/95235. 72. Gigante B, Santos C, Silva AM, Curto MJM, Nascimento
66. Pinelo M, Rubilar M, Sineiro J, Núñez MJ. Extraction of MSJ, Pinto E, Pedro M, Cerqueira F, Pinto M. M, Duarte M
antioxidant phenolics from almond hulls (Prunus P, Laires A, Rueff J, Gonçalves J, Pegado M I, Valdeira M
amygdalus) and pine sawdust (Pinus pinaster). Food Chem. L. Catechols from abietic acid synthesis and evaluation as
2004; 85(2):267‑73. bioactive compounds. Bioorg Med Chem. 2003;
https://doi.org/10.1016/j.foodchem.2003.06.020. 11(8):1631‑8. https://doi.org/10.1016/S0968-
67. Hayet E, Maha M, Samia A, Ali MM, Souhir B, Abderaouf 0896(03)00063-4.
K, Mighri Z, Mahjoub A. Antibacterial, antioxidant and 73. Elliger CA, Zinkel DF, Chan BG, Waiss AC. Diterpene acids
cytotoxic activities of extracts of Conyza canadensis (L.) as larval growth inhibitors. Experientia. 1976;32(11):1364‑6.
Cronquist growing in Tunisia. Med Chem Res. 2009; https://doi.org/10.1007/BF01937376.
18(6):447‑54. https://doi.org/10.1007/s00044-008-9141-0. 74. Dzeha T, Jaspars M, Tabudravu J. Clionasterol, a
68. Todorovic V, Redovnikovic IR, Todorovic Z, Jankovic G, Triterpenoid from the Kenyan Marine Green Macroalga
Dodevska M, Sobajic S. Polyphenols, methylxanthines, and Halimeda macroloba. West Ind Oc J Mar Sci. 2004;
antioxidant capacity of chocolates produced in Serbia. J Food 2(2):157‑61.
Compost Anal. 2015; 41:137‑43. 75. Khan ZM Halleys, Hossain Md Faruquee, Md Munan Shaik.
https://doi.org/10.1016/j.jfca.2015.01.018. Phytochemistry and Pharmacological Potential of
69. Bérubé-Gagnon J. Isolation et identification de composés Terminalia arjuna L. J. Med. Plant Res. 2013; 3(10):70-77.
antibiotiques des écorces de Picea mariana / Université du doi:10.5376/mpr.2013.03.0010.
Québec à Chicoutimi [Online]. 2006 [cited 2023 Jan 12]. 76. Serrano G, Fortea JM, Millan F, Botella R, Latasa JM.
Available from: http://constellation.uqac.ca/449/. Contact allergy to resorcinol in acne medications: report of
70. Söderberg TA, Gref R, Holm S, Elmros T, Hallmans G. three cases. J Am Acad Dermatol. 1992; 26(3):502‑4.
Antibacterial Activity of Rosin and Resin Acids in Vitro. https://doi.org/10.1016/S0190-9622(08)80588-9.
Scand. J. Plast. Reconstr. Surg. Hand Surg. 1990;

6750
© 2023 the authors. This work is licensed under the Creative Commons Attribution 4.0 International License