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205
July 2022. Volume 8. Number 3
Samaneh Rahamouz Haghighi1* , Alireza Yazdinezhad2 , Khadijeh Bagheri1, Ali Shara3,4*
1. Department of Plant Production and Genetics, Faculty of Agriculture, University of Zanjan, Zanjan, Iran.
2. Department of Pharmacognosy, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
3. Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran.
4. Department of Pharmaceutical Biotechnology, School of Pharmacy, Zanjan University of Medical Sciences, Zanjan, Iran.
Original Article
Volatile Conituents and Toxicity of Essential Oils Extract-
ed From Aerial Parts of Plantago Lanceolata and Plantago
Major Growing in Iran
Background: Plantago lanceolata L. (P. lanceolate) and Plantago major L. (P. major) belong
to the Plantaginaceae family and are widely used in traditional medicine.
Objectives: This udy aims to qualitatively identify the crucial compounds and evaluate the
toxicity eects of essential oils of two Plantago species.
Methods: The plantains were collected from Zanjan Province, Iran. The essential oils were
extracted by hydrodiillation and then analyzed using gas chromatography coupled with mass
spectrometry (GC/MS). The toxicity eects of the essential oils were evaluated on HCT-116
and HEK-293 cell lines (in vitro MTT assay) and Artemia salina (A.salina) (in vivo assay).
The conituents of the essential oils were identied by calculating their retention indices
under temperature-programmed conditions for n-alkanes (C8-C20) in the Agilent 19091S-433
column.
Results: The main identied conituents were metaraminol (14.04%), bifemelane
(8.73%), metossamina (8.16%), and pterin-6-carboxylic acid (5.11%) in P. lanceolata and
2-dodecen-1-yl (-) succinic anhydride (15.29%), benzenemethanol, α-(1-aminoethyl)-2,5-
dimethoxy-(11.83%), dl-phenylephrine (7.51%), and nortriptyline (5.15%) in P. major. The
essential oils of P. major exhibited more antiproliferative properties on HCT-116 at 72 h
compared to P. lanceolata (IC50: 102.66 µg/mL). At 400 µg/mL of P. lanceolata and P. major,
the percentage of the lethality of nauplii was 8% and 12%, respectively (LC50:2242.57 µg/
mL and 1783.7 µg/mL). The present udy showed that the mo of conituents of oils were
alcohols and amines.
Conclusion: Some of the compounds identied in the Plantago species essential oils have
important pharmaceutical properties. This udy reported the cytotoxicity of essential oils on
the colon cancer cell line. However, the essential oils were not toxic again A.salina at the
examined concentrations.
A B S T R A C T
Keywords:
Brine shrimp, Colorectal
cancer, Plantago
lanceolata L., Plantago
major L. Volatile oils
Article info:
Received: 13 Jul 2022
Accepted: 12 Feb 2022
Copyright© 2020, The Authors.
* Corresponding Author:
Samaneh Rahamouz Haghighi, PhD.
Address: Department of Plant Production and Genetics, Faculty of Agri-
culture, University of Zanjan, Zanjan, Iran.
Phone: +98 (91) 51236427
E-mail: rahamouz_haghighi.s@yahoo.com
Ali Shara, PhD.
Address: Zanjan Pharmaceutical Biotechnology Research Center, Zanjan
University of Medical Sciences, Zanjan, Iran.
Phone: +98 (24) 33473635
E-mail: Shara.a@gmail.com
Citation
Rahamouz Haghighi S, Yazdinezhad A, Bagheri K, Shara A. Volatile Conituents and Toxicity of Essential Oils Extracted From Aerial Parts
of Plantago Lanceolata and Plantago Major Growing in Iran. Pharmaceutical and Biomedical Research. 2022; 8(3):205-224.
:
: http://dx.doi.org/10
206
July 2022. Volume 8. Number 3
Introduction
ssential oils are used as additives in many
types of foods and beverages and various
food supplements [1]. The Plantago genus of
the Plantaginaceae family includes approxi-
mately 300 annual and perennial species,
growing worldwide, and specially cultivated
in the subtropical regions [2]. According to Iran’s tradition-
al medicine, Plantago species have many medical applica-
tions without serious side eects; however, some of the
medicinal eects of Plantago lanceolata L. (P. lanceolata)
and Plantago major L. (P.major) in Iran’s traditional medi-
cine have not been discovered in modern medicine [3].
P. lanceolata and P. major are used to treat wounds,
infectious diseases, digeive and respiratory problems,
fever, pain, dermatitis, and tumors [4, 5]. Furthermore,
Plantago species were used to cure burns, ulcers, and eye
diseases, as anti-inammatory, antipyretic agents, anti-
tussive, and purgative for snakebites [6]. Researchers
have also reported that P.major mucilage can optimize
the drug release in propranolol buccoadhesive tablets
[7]. Additionally, they can be used in cosmetics to pro-
duce face masks, creams, or lotions for acne-prone and
oily skins because of their aringent, anti-septic, and
anti-bacterial properties [6].
GC/MS is one of the mo important inruments used to
analyze a sample with volatile conituents as it combines
both the chromatographic technique for the ecient sepa-
ration of sample conituents and mass spectroscopy that
identies the compounds according to their mass-to-charge
ratio (m/z) [8]. The above-mentioned properties of these
plants provide us with signicant reasons to analyze their
volatile composition. To date, only a few Plantago species
have been inveigated for their chemical conituents and
biological activities of extracts. Previous udies on the
chemical inveigation of Plantago L. leaves and seeds ex-
tracts demonrated the presence of polysaccharides, phe-
nolic acids, avonoids, iridoid glycosides, and vitamins [2].
There are few valid udies on the essential oil com-
positions of P. lanceolata and P. major, considering
that these plants contain very small amounts of es-
sential oil. Therefore, in the current udy, following
our previous udies on these plants, their essential oil
compositions were examined. In addition, we evalu-
ated the toxicity eects of the essential oils on colon
cancer cells and Artemia salina (A.salina). To the be
of our knowledge, there are no reports on the cytotox-
icity assay of P. lanceolata and P. major essential oils
on colon cancer cell lines.
Materials and Methods
Herbal material
The aerial parts (leaf and em) of P. lanceolata and
P. major were collected from Zanjan Province, Iran (the
geographical coordinates of the collection sites are as
follows: 36°41’15.5”N 48°24’02.2”E). The taxonomic
identity of species was authenticated at the Department
of Botany, University of Zanjan, Iran. All sections were
cut into small pieces and were dried in shade and at room
temperature separately for one week.
Isolation of essential oils
The aerial parts of P. lanceolata and P. major (100 g)
were ground to a coarse powder and extracted with 1500
mL of diilled water for hydrodiillation in a Cleveng-
er-type apparatus for 5 to 6 h to arise the volatile compo-
sition in the form of essential oils. The essential oils were
collected into 1 mL of n-pentane and then poured into a
glass and ored at 4°C until further analysis [1].
Gas chromatography-mass spectrometry analysis
The essential oils of the aerial parts of P. lanceolata
and P. major were used for GC/MS analysis. GC/MS
analysis was performed using the Agilent technologies
5975c. GC/MS analysis was carried out by 1 µL of the
materials subjected to analysis. The GC/MS syem
has been equipped with a capillary column (30 m×250
µm×0.25 µm, Agilent). Helium as the carrier gas was
used at the ow rate of (1 mL/min). The injector and
the interface temperature were maintained at 250°C.
The column temperature was programmed as follows:
the initial temperature was 40°C (1 min) and then it in-
creased at a rate of 2°C/min up to 200°C (10 min). The
identication of the conituents of P. lanceolata and P.
major was performed by comparison with MS literature
data (NIST08.L) and retention index (RI) [1]. The mix-
tures of n-alkanes (C8-C20) were injected using the above
temperature program to calculate the RI for each peak.
The RI of the compounds was calculated using the fol-
lowing equation:
1.
Where: (Ix) is the Kovats retention index; (n) is the
number of carbon atoms in the alkane; (tn) and (tn+1)
are the retention times of the reference n-alkane hydro-
E
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
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July 2022. Volume 8. Number 3
carbons with n and n + 1 carbon atoms; and (tx) is the
retention time of the peak of the unknown compound.
Several peaks did not have RIs for the calculated mix-
tures of n-alkanes (C8-C20). Thus, compounds with a
formula ructure less than C8 and more than C20 could
not be calculated (these compounds were considered un-
known).
Cell line culture
Human embryonic kidney cell (HEK-293) as a nor-
mal cell line and colorectal cancer cell line (HCT-116)
provided by the Paeur Initute of Iran, Tehran were
cultured in the Dulbecco’s Modied Eagle Medium with
supplementation of penicillin-reptomycin (1%) along
with 10% fetal bovine serum incubated in 5% CO2 in-
cubator at 37°C.
Cytotoxicity assay
The MTT assay was performed to evaluate the cyto-
toxicity of P. lanceolata and P. major essential oils on
the cell lines [9]. A 96-well plate with a density of 7 ×
103 cells/well were used for cell seeding. The cells were
allowed to attach and grow for 24 h. The cells under-
went treatment with 25-400 µg/mL concentrations. The
HCT-116 were treated with 5-uorouracil (5-FU) (Aus-
tria, Ebewe Pharma) in dierent doses (2.5-10 μg/mL)
for 72 h. The 5-FU and untreated cells were utilized as
the positive and negative control, respectively. The ad-
dition and incubation of 20 μL of MTT (5 mg/mL) for
4 h took place after 24 to 72 h, followed by removing
the medium and adding 200 μL of dimethyl sulfoxide to
dissolve the obtained formazan. An ELISA plate reader
(Tecan Innite M200, Auria) at 570 and 690 nm read
the absorbance. The cell growth inhibition rates were ex-
amined by the following formula:
2.
Where: (A) indicates the absorbance.
Toxicity assay on artemia salina
The larvae of brine shrimp (A.salina Leach) were em-
ployed to examine the P. lanceolata and P. major es-
sential oils’ overall toxicity [10]. A. salina eggs were
provided by Urmia University, the We Azerbaijan
Province, Iran. A ask with 35 g of NaCl dissolved in 1
L of diilled water was used for cy culture, followed
by 48 h incubation at 28°C and the larvae hatching after
48 h. Every well in the 96-well microtiter plates hav-
ing the Roswell Park Memorial Initute (RPMI-1640)
received the essential oils (25-400 µg/mL). The next
ep included the addition of 10 nauplii per well to the
96-well plates and incubation at a temperature of 25°C
for 24 h. A binocular microscope was employed to cal-
culate the number of live nauplii in every well after 24
h. All experiments were repeated 3 times. Additionally,
the negative control contained only 10 nauplii and ar-
ticial seawater. Potassium dichromate (K2Cr2O7) was
used as a positive control at the same concentrations as
the essential oils. The number of survived samples in the
experimental and control wells was used to calculate the
percentages of the nauplii morality. The Abbott formula
determined the lethality:
Statistical analysis
The data were analyzed using the SPSS software, ver-
sion 21. The signicant dierences between means were
calculated. Values were expressed as the mean of the 3
replications ± Standard Deviation (SD). The Duncan te
at P value<0.05 was used to determine signicant dier-
ences among treatments. IC50 and LC50 values were ana-
lyzed with the ED50 plus v1.0 Software.
Results
Many peaks were detected in the chromatogram of the
essential oils extracted from P. lanceolata and P. ma-
jor aerial parts by GC/MS and their compositions were
identied according to the NIST08.L library. Figure 1
shows the main chromatograms of the essential oils of
P. lanceolata and P. major. The essential oils were rich
in amine derivations, alcohols, alkenes, and fatty acids.
The essential oils also showed the presence of acids, al-
kaloids, amino acids, carboxylic acid derivatives, eers,
ketones, monoterpenoids, nitriles, oximes, phenols,
phenethylamine derivatives, and others (Table 1).
Volatile constituents of P. lanceolata essential oil
Mo component of P. lanceolata essential oil is gen-
erated by metaraminol (14.04%), bifemelane (8.73%),
metossamina (8.16%), and pterin-6-carboxylic acid
(5.11%).
In the present udy, 106 components belonging to
main chemical groups were identied in P. lanceo-
lata essential oil: alcohols (17.56%) with benzyl alco-
hol; .α.-(1-aminoethyl)-m-hydroxy-, (-)-(14.04) as the
main component; amines (14.70%) with phenylephrine
(3.71%); alkenes and alkenes (12.28%) with bifemelane
(8.73%); ketones (8.70%) with bicyclo [2.2.1] heptan-
2-one, 4,7,7-trimethyl-, semicarbazone (2.97%); acids
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
208
July 2022. Volume 8. Number 3
(8.05%) with pterin-6-carboxylic acid (5.11%); alka-
loids (5.76%) with 2H-1,2,3-triazole-4-carboxylic acid;
2-(2-uorophenyl)- (2.12%); eers (4.02) with 2-thio-
pheneacetic acid; 3,5-diuorophenyl eer (1.53%); am-
ides (3.55%) with propanamide (0.58%); amino acids
(2.71%) with hiidine; 1, N-dimethyl-4-nitro- (1.76%);
monoterpenoids (2.45%) with Linalool (0.97%); phenol
(Benzeneethanamine, 2-uoro-.beta.,5-dihydroxy-N-
methyl-) (0.45%); nitriles (0.21%) with propanenitrile,
3-(methylamino)- (0.17%); oximes with ethanone,
1-(4-pyridinyl)-, oxime (0.13%) as the main components
and others (21.03%) (Table 2 and 3). The biological ac-
tivities of the volatile conituents of P. lanceolata oil are
reported in Table 4.
Volatile constituents of the essential oils of p. major
The present udy showed that 2-dodecen-1-yl (-)
succinic anhydride (15.29%), benzenemethanol,. α.-(1-
aminoethyl)-2,5-dimethoxy- (11.83%), dl-phenyleph-
rine (7.51%), nortriptyline (5.15%) were the major con-
ituents (Tables 2 and 3).
In the present udy, 79 components belonging to main
chemical groups were identied in P. major essential oil:
amines (35.74%) with phenylephrine (11.66%) as the
main component; alkenes and alkanes (24.88%) with
2-dodecen-1-yl(-)succinic anhydride (15.29%); phenols
(10.49%) with dl-phenylephrine (7.51%); eers (6.96%)
with sarcosine, N-valeryl-, butyl eer (2.02%); alcohols
(5.14%) with cyclobutanol, 2-ethyl- (1.72%); alkaloids
(3.97%) with ethylamine, 2-(adamantan-1-yl)-1-meth-
yl- (0.28%); ketones (3.61%) with 3-(E)-hexen-2-one,
(5S)-5-[(t-butoxycarbonyl-(R)-alanyl)amino]- (2.65%);
amides (2.2%) with [(2,5-dimethoxyphenyl)sulfonyl]
ethylamine (0.69%); monoterpenes with isoborneol
(1.17%); amino acids (glycine, N-(N-L-alanylglycyl)-)
(0.35%) and acid (0.16%) with imidazole-5-carboxylic
acid, 2-amino- as the main component. P.major essential
oil has many properties and applications that are pro-
vided in Table 4.
The essential oils of P. lanceolata and P. major spe-
cies showed that the predominant compounds were
present in both species; however, the amounts of these
compounds (%) were dierent. For example, (-)-Ben-
zyl alcohol, .α.-(1-aminoethyl)-m-hydroxy (14.04%
and 1.37%), metossamina (8.16% and 0.17%), benzen-
emethanol, .α.- (1-aminoethyl) -2,5-dimethoxy- (3.71%
and 11.66%), dl-phenylephrine (0.15% and 7.51%), nor-
triptyline (0.95% and 5.15%) were present in P. lanceo-
lata and P. major, respectively (Figure 2). Bifemelane
(% 8.73), pterin-6-carboxylic acid (5.11%) exied only
in P. lanceolata while 2-dodecen-1-yl (-) succinic anhy-
dride (15.29%) were only found in P. major.
Cytotoxic activities
Table 1. Major compound groups obtained from extracted essential oil of plantago lanceolata and plantago major aerial parts
Classicaon of Composions Plantago Lanceolata (%) Plantago Major (%)
Alcohols 17.5694 5.14
Alkaloids 5.7652 3.97
Alkanes and alkenes 12.2893 24.88
Amides 3.5522 2.2
Amines 14.7012 35.74
Amino acids 2.711 0.35
Esters 4.0211 6.96
Ketones 8.7041 3.61
Phenols 0.4593 10.49
Terpenes 2.4556 1.17
Others 29.4376 7.09
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
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July 2022. Volume 8. Number 3
Table 2. Identied compositions in plantago lanceolata essential oil by hydrodistillation
RI Area Pct
(%) Library/ID – (Plantago Lanceolata)Formula Molecular
Weight RI Area Pct
(%) Library/ID – (Plantago Major)Formula Molecular
Weight
1230.29 0.043 Uramil-N,N-diacec acid C8H9N3O7259.17 1352.44 1.37 Benzyl alcohol, .alpha.-(1-aminoethyl)-m-hydroxy-, (-)- C9H13NO2167.20
1234.07 0.2343 Phosphonic acid, (1-aminoethyl)-,
bis(trimethylsilyl) ester C8H24NO3PSi2269.43 1354.88 0.85 Benzeneethanamine, 4-chloro-.alpha.-methyl- C9H12CIN 169.65
1251.93 0.126 Adrenalone C9H11NO3181.19 1358.38 0.13 1,2-Benzenediol, 4-[2-(methylamino)ethyl]- C9H13NO2167.2
1262.13 0.0875 1-Methyl-2-phenoxyethylamine C9H13NO 151.21 1365.88 1.8 Benzeneethanamine, 2-uoro-.beta.,5-dihydroxy-N-
methyl- C9H12FNO2185
1273.40 0.9972 Quinoline, 4-methyl-, 1-oxide C10H9NO 159.18 1368.89 0.43 Phenethylamine, p,.alpha.-dimethyl- C10H15N149.23
1290.69 0.1024 2-Amino-1-(o-methoxyphenyl)propane C10H15NO 165.2322 1379.10 0.79 Epinephrine C9H13NO3183.2
1292.05 1.1679 [2,7]Naphthyridine-1,3,6,8-tetraol C8H6N2O4194.14 1380.34 0.28 Benzeneethanamine, N-methyl- C9H13N135.2
1296.54 0.0628 1,2-Benzenediol, 4-(2-amino-1-hydroxy-
propyl)- C9H13NO3183.2 1384.20 0.15 2-(5-Methylaminopentyl)-5-methylthio-1,3,4-thiadia-
zole C9H17N3S2231.4
1307.43 1.761 Hisdine, 1,N-dimethyl-4-nitro- C8H12N4O4228.21 1384.98 0.19 Phenol, 4-(2-aminopropyl)- C9H13NO 151.21
1319.98 2.1206 2H-1,2,3-Triazole-4-carboxylic acid,
2-(2-uorophenyl)- C9H6FN3O2207.16 1397.16 0.93 2-Amino-1-(o-methoxyphenyl)propane C10H15NO 165.23
1330.54 0.6905 Phenylpropanolamine C9H13NO 151.21 1424.89 0.39 3,4-Methylenedioxy-amphetamine C10H13NO2179.22
1345.22 0.2952 l-Alanine, N-(1-oxopentyl)-, methyl ester C9H17NO3187.24 1457.00 0.19 Propanamide, N-(1-cyclohexylethyl)- C11H21NO 183.29
1346.19 0.1483 dl-Phenylephrine C9H13NO2167.2 1466.05 1.26 3-Methoxyamphetamine C10H15NO 165.23
1353.01 0.9611 Racepinephrine C9H13NO3183.2 1468.92 2.68 Phenethylamine, p-methoxy-.alpha.-methyl-, (.+/-.)- C10H15NO 165.23
1356.10 0.0546 2-(5-Aminohexyl)furan C10H17NO 167.25 1479.59 0.17 Benzeneethanamine, 3,4-dimethoxy-N-methyl- C11H17NO2195.62
1362.62 0.4638 Epinephrine C9H13NO3183.2 1481.73 0.32 Metanephrine C10H15NO3197.23
1366.25 0.4593 Benzeneethanamine, 2-uoro-.beta.,5-
dihydroxy-N-methyl- C9H12FNO2185.2 1514.77 0.54 3-Buten-2-one, 4-(2,5,6,6-tetramethyl-1-cyclohexen-
1-yl)- C14H22O206.32
1368.84 0.0522 Metanephrine C10H15NO3197.23 1522.72 0.3 Mexilene C11H17NO 179.25
1375.33 3.7122 Benzenemethanol, 3-hydroxy-.alpha.-
[(methylamino)methyl]-, (R)- C9H13NO2167.205 1548.55 0.72 3-Ethoxyamphetamine C11H17NO 179.26
1382.87 0.301 2-Buten-1-one, 1-(2,6,6-trimethyl-1,3-
cyclohexadien-1-yl)- C13H18O190.2814 1551.58 0.06 2-Ethoxyamphetamine C11H17NO 179.26
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
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July 2022. Volume 8. Number 3
RI Area Pct
(%) Library/ID – (Plantago Lanceolata)Formula Molecular
Weight RI Area Pct
(%) Library/ID – (Plantago Major)Formula Molecular
Weight
1390.68 2.1026 m-Menth-1(7)-ene, (R)-(-)- C10H18 138.25 1576.31 0.12 Benzeneethanamine, 3,4-dimethoxy-.alpha.-methyl- C11H17NO2195.26
1408.33 0.2024 Sarcosine, N-valeryl-, ethyl ester C10H19NO3201.26 1587.37 0.55 2-Hexanamine, 5-methyl- C7H17N115.22
1431.35 0.2885 8-Azabicyclo[4.3.1]decan-10-one, 8-methyl- C10H17NO 167.25 1612.19 1.06 1-(2-Cyano-2-ethyl-butyryl)-3-isopropyl-urea C11H19N3O2225.29
1439.41 0.0497 N-2,4-Dnp-L-arginine C12H16N6O6340.29 1616.34 0.17 Benzenemethanol, .alpha.-(1-aminoethyl)-2,5-dime-
thoxy- C11H17NO3211.26
1453.15 0.0837 N-Isopropyl-3-phenylpropanamide C12H17NO 191.27 1707.88 0.44 2,5-Dimethoxy-4-(methylthionyl)amphetamine C12H19NO3S257.35
1456.58 1.7548 8-Amino-6-methoxyquinoline C10H10N2O174.2 1749.06 2.02 Sarcosine, N-valeryl-, butyl ester C12H23NO3229.31
1457.67 0.1953 Tocainide C11H16N2O192.26 1766.30 0.13 Acnobolin C13H20N2O6300.31
1465.31 0.9648 L-Asparc acid, N-(2,4-dinitrophenyl)- C10H9N3O8299.19 1786.65 0.11 2-(2-N-Methylaminoethyl)-4-hydroxy-5-methoxyphen-
ylacecacid, methyl ester C13H19NO4253.29
1494.26 0.8048 3,5-Dimethylamphetamine C11H17N163.26 1787.68 3.52 2-(3-Phenyl-piperidin-1-yl)-ethylamine C13H20N2204.31
1494.91 0.2261 Tricyclo[4.3.1.1(3,8)]undecane-1-carboxylic
acid C12H18O2194.27 1813.29 1.24 Sarcosine, N-valeryl-, pentyl ester C13H25NO3243.34
1526.32 0.4584 Benzenepropanoic acid, .alpha.-(1-amino-
ethyl)-, [R-(R*,R*)]- C11H15NO2193.24 1826.57 0.93 5-Isoxazolepropanamine, N-methyl-3-(4-nitrophenyl)- C13H15N3O3
1532.76 0.1697 Benzeneethanamine, 2-uoro-.beta.-
hydroxy-4,5-methoxy-.alpha.-methyl- C11H16FNO3229.25 1900.78 1.74 Sarcosine, N-valeryl-, isohexyl ester C14H27NO3257.37
1542.60 2.9722 Bicyclo[2.2.1]heptan-2-one, 4,7,7-tri-
methyl-, semicarbazone C11H19N3O209.29 1915.13 5.15 Nortriptyline C19H21N263.38
1573.85 0.3682 (2-Indol-1-yl-ethyl)-methyl-amine C11H14N2174.24 1916.40 0.73 1-[.alpha.-(1-Adamantyl)benzylidene]thiosemicarba-
zide C18H23N3S313.5
1592.46 8.1614 Benzenemethanol, .alpha.-(1-
aminoethyl)-2,5-dimethoxy- C11H17NO3211.26 2032.74 0.8 Benzeneethanamine, .alpha.-methyl-3-[4-methyl-
phenyloxy]- C16H19NO 241.32
1598.97 0.5233 Propanamide, 3-(3,4-dimethylphenylsul-
fonyl)- C11H15NO3S241.31 2053.33 0.06 Ethanamine, N-methyl-2-[(2-methylphenyl)phenyl-
methoxy]- C17H21NO 255.35
1633.16 0.1986 Acetamide, 2-(adamantan-1-yl)-N-(1-ada-
mantan-1-ylethyl)- C14H22ClNO 355.6 2150.11 0.21 l-Alanine, N-octanoyl-, pentyl ester C16H31NO3285.42
1633.68 0.3392 Folic Acid C19H19N7O6441.4 2154.26 0.47 Desmethyldoxepin C18H19NO 265.3
1639.54 0.3155 3-Propoxyamphetamine C12H19NO 193.28 2194.96 1.56 1-Octadecanamine, N-methyl- C19H41N283.53
1699.53 1.3231 3-Methyl-3,5--(cyanoethyl)tetrahydro-
4-thiopyranone C12H16N2OS 2198.92 0.26 1-Methyl-4-[nitromethyl]-4-piperidinol C7H14N2O3174.2
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RI Area Pct
(%) Library/ID – (Plantago Lanceolata)Formula Molecular
Weight RI Area Pct
(%) Library/ID – (Plantago Major)Formula Molecular
Weight
1732.19 0.2144 l-Alanine, N-capryloyl-, methyl ester C12H23NO3229.3159 2411.60 0.42 3,3-Dimethyl-4-methylamino-butan-2-one C7H15NO 129.2
1831.57 0.4466 2-(2-N-Methylaminoethyl)-4-hydroxy-5-me-
thoxyphenylacecacid, methyl ester C13H19NO4253.29 2482.03 0.35 Glycine, N-(N-L-alanylglycyl)- C7H13N3O4203.19
1833.98 1.0636 5-Isoxazolepropanamine, N-methyl-3-(4-
nitrophenyl)- C13H15N3O3261.28
1915.13 0.9451 Nortriptyline C19H21N263.4
1943.73 0.6324 Benzeneethanamine, .alpha.-methyl-3-[4-
methylphenyloxy]- C16H19NO 241.33
1963.23 0.2011 2,5-Dimethoxy-4-propylamphetamine C14H23NO2237.34
1993.21 0.4336 Benzofuran-5-ol, 3-(2-furanoyl)-4-dimethyl-
aminomethyl- C16H15NO4285.29
2051.87 0.2945 3,3-Dimethyl-4-methylamino-butan-2-one C7H15NO 129.2
2075.78 2.9079 Atomoxene C17H21NO 255.35
2092.67 0.5213 8-Methyl-2,3,3a,4,5,6-hexahydro-1H-
pyrazino[3,2,1-jk]carbazole-3-carboxamide C16H19N3O269.34
2120.85 1.0019 Desmethyldoxepin C18H19NO 265.3
2137.14 0.2377 Pentanamide, N-decyl-N-methyl- C16H33NO 255.44
2163.98 1.2397 2-(4,5-Dihydro-3-methyl-5-oxo-1-phenyl-
4-pyrazolyl)-5-nitrobenzoic acid C17H13N5O5367.3
2176.19 8.7366 Bifemelane C18H23NO 269.4
2226.56 0.4172 Northiaden C18H19NS 281.4
2236.63 0.053 1-Methyl-4-[nitromethyl]-4-piperidinol C7H14N2O3174.2
2420.85 0.7689 Ethyl isopropyl dimethylphosphoramidate C7H18NO3P195.2
2452.66 5.1189 Pterin-6-carboxylic acid C7H5N5O3207.15
Notes: Non-isothermal Kovats retention indices (from temperature-programming, using denition of Van den Dool and Kratz); RI, retention index on Agilent 19091S-433.
Ix=100n+100[log(tx)-log(tn)]/[log(tn+1)-log(tn)]; (n), the number of carbon atoms in the alkane; (tn) and (tn+1), the retention times of the reference n-alkane hydrocarbons with n and n +
1 carbon atoms; tx, retention time of peak of unknown compound.
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Table 3. Unidentied compositions in plantago lanceolata essential oils by hydrodistillation
RT Area Pct Library/ID – (Plantago Lanceolata)Formula Molecular
Weight RT Area
Pct Library/ID – (Plantago Major)Formula Molecular
Weight
3.1401 0.0989 Sarcosine, n-hexanoyl-, pentadecyl ester C24H47NO3397.6 3.15 15.29 2-Dodecen-1-yl(-)succinic anhydride C16H26O3266.38
3.5759 0.1056 Cyclopropanecarboxamide C4H7NO 85.1 13.21 1.72 Cyclobutanol, 2-ethyl- C6H12O100.16
3.9266 0.0734 1-[alpha-(1-Adamantyl)benzylidene]thiosemicarbazide nd nd 20.74 1.56 Thiophene-3-ol, tetrahydro-, 1,1-dioxide C4H8O3S136.17
4.2667 0.1183 Benzenemethanol, alpha-(1-aminoethyl)-, (R*,R*)- nd nd 21.03 0.47 Cyclopropyl carbinol C4H8O72.1
4.3199 0.1706 Propanenitrile, 3-(methylamino)- C4H8N284.12 26.67 0.33 Acetamide, 2-chloro- C2H4ClNO 93.512
6.977 0.0332 L-Alanine, methyl ester C4H9NO2103.12 50.75 0.43 2-Amino-1-(o-hydroxyphenyl)propane - -
7.7211 0.0328 2-Isopropoxyethylamine C5H13NO 103.16 60.24 0.28 Ethylamine, 2-(adamantan-1-yl)-1-methyl- - -
20.8793 0.0445 Propanenitrile, 3-amino-2,3-di(hydroxymino)- C3H4N4O2128.09 60.74 0.83 4-Fluorohistamine C5H8FN3129.13
40.2128 0.2184 2,4-Dimethylamphetamine nd nd 64.27 0.75 Cycloserine C3H6N2O2102.09
43.4439 0.0565 Adipamide C6H12N2O2144.17 64.97 0.13 3-Hydroxy-N-methylphenethylamine - -
45.0488 0.423 4-Fluorohistamine C5H8FN3129 72.23 0.29 Acetamide, 2,2-dichloro- C2H3CL2NO 127.95
47.7166 0.2813 Acetamide, 2-cyano- C3H4N2O84.08 74.16 0.26 Propanamide C3H7NO
52.7971 0.081 2-Bromoacetamide C2H4BrNO 137.96 75.99 0.23 dl-3-Aminoisobutyric acid, N-methyl-, methyl ester C6H13NO2131.17
55.5605 0.0734 Carbamic acid, N-[(N-cyanomethylpropanamide)-2-yl]-,
1-methyl-1-(3,5-dimethoxyphenyl)ethyl ester nd nd 76.18 0.34 2,2-Dichlorocyclopropanecarboxamide C4H5CL2NO 153.99
56.4108 0.3166 Cyanoacetylurea C4H5N3O2127.1 77.09 1.87 Methylpent-4-enylamine C6H13N99.17
56.8572 0.3047 Propan-1-one, 2-amino-1-piperidin-1-yl- nd nd 77.84 0.24 1,4-Benzenedicarboxamide, N,N’-bis(2-hydroxy-1-methyl-
2-phenylethyl)- C26H28N2O432.5
57.4205 0.2208 Acetamide, 2,2,2-trichloro- C2H2Cl3NO 162.4 77.96 0.16 4H-1,3-Dioxino[5,4-c]pyridine, hexahydro-6-methyl-8a-
phenyl-
57.8138 0.0713 2,4-Bis(hydroxylamino)-5-nitropyrimidine C4H5N5O4187.11 79.54 0.19 2,4-Bis(hydroxylamino)-5-nitropyrimidine C4H5N5O4187.11
62.3629 0.9949 Propylamine, 3-(furan-2-yl)-1-methyl- nd nd 80.13 0.16 Imidazole-5-carboxylic acid, 2-amino-
62.6817 1.9931 3-Chloro-N-methylpropylamine C4H10ClN 107.58 83.05 2.65 3-(E)-Hexen-2-one, (5S)-5-[(t-butoxycarbonyl-(R)-alanyl)
amino]- C14H24N2O4284.35
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RT Area Pct Library/ID – (Plantago Lanceolata)Formula Molecular
Weight RT Area
Pct Library/ID – (Plantago Major)Formula Molecular
Weight
63.6064 0.7875
3,6-Methano-8H-1,5,7-trioxacyclopenta[ij]cycloprop[a]
azulene-4,8(3H)-dione, hexahydro-9-hydroxy-8b-methyl-
9-(1-methylethyl)-, [1aR-(1a.alpha.,2a.beta.,3.beta.,6.
beta.,6a.beta.,8aS*,8b.beta.,9R*)]-
nd nd 84.87 0.82 l-Alanine, N-octanoyl-, decyl ester C21H41NO3355.6
63.7127 0.562 N-(3-Methylaminopropyl)-N-methylformamide C6H14N2O 130.19 86.2 0.2 Acetamide, 2,2,2-trichloro- C2H2Cl3NO 162.4
64.5949 0.5809 Propanamide C3H7NO 73.09 87.53 0.69 N-Ethyl-2,5-dimethoxy-benzenesulfonamide - -
65.1157 0.2673 Benzenemethanol, .alpha.-(1-aminoethyl)-, (R*,R*)-(.+/-.)- nd nd 88.87 0.59 l-Alanine, N-valeryl-, tridecyl ester C21H41NO3355.6
67.5922 3.9122 Imidazole, 2-amino-5-[(2-carboxy)vinyl]- C6H7N3O2153.14
70.7489 0.294 2-Propen-1-amine, 2-bromo-N-methyl- C4H8BrN 150.02
72.4282 0.505 2-Methylaminomethyl-1,3-dioxolane C5H11NO2117.15
72.5026 0.536 Methanesulfonamide, N,N-dimethyl- C3H9NO2S123.174
74.5645 0.1525 Sarcosine, N-valeryl-, butyl ester nd nd
75.5636 0.2446 Pyridine-3-carboxamide, 1,2-dihydro-4,6-dimethyl-
2-thioxo- nd nd
76.4777 0.7274 Benzyl alcohol, p-hydroxy-.alpha.-[(methylamino)methyl]- nd nd
78.7841 0.2749 8-[N-Aziridylethylamino]-2,6-dimethyloctene-2 C13H20O192.3
80.7079 1.6954 Phenol, 4-(2-aminopropyl)-, (.+/-.)- C9H13NO 151.21
81.4307 2.4441 3-(E)-Hexen-2-one, (5S)-5-[(t-butoxycarbonyl-(S)-alanyl)
amino]- C14H24N2O4284.35
83.5032 14.0414 Benzyl alcohol, alpha.-(1-aminoethyl)-m-hydroxy-, (-)- nd nd
88.4243 1.5345 2-Thiopheneacec acid, 3,5-diuorophenyl ester nd nd
Notes: These compounds were obtained from the NIST08.L library and identied by CAS numbers.ثأ
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Table 4. Biological Activities of Volatile Compositions of Plantago Lanceolata and Plantago Major
Library/ID – (Plan-
tago Lanceolata)Biological Acvity Structure Library/ID –
(Plantago Major)Biological Acvity Structure
1,6-Octadien-3-ol,
3,7-dimethyl- or
Linalool
An-inammatory, an-cancer acvies [11] 2-Dodecen-1-yl(-)
succinic anhydride
An-convulsant, an-neoplasc agents, an-
oxidants, an-microbial acvies [12]
1-Octen-3-ol
A strong an-bacterial, inhibion of the growth
of insects [13], a profound inuence on protein
expression paerns, blocking isotropic growth,
mild physiological eects on germinang conidia
in soluon [14]
Phenylephrine Alpha-adrenergic agonist, decongestant, an-
bacterial acvity [15]
2-Furanmethanol,
5-ethenyltetrahydro-.
alpha.,.alpha.,5-
trimethyl-, cis
An-viral, an-oxidave acvies [16] 2-Chloroacetamide An-microbial agent, [17] herbicides [18]
2-Isopropoxyethyl-
amine An-microbial acvity [19] 2,5-Norbornadiene To block the ethylene receptor of plant
ssues [20]
8-Amino-6-methoxy-
quinoline An-malaria acvity [21] Isoborneol An-viral, [22] anbacterial eects, [23] an-
bacterial acvies [24]
Arginine An-microbial acvity [25] 1-Methyldecylamine Inseccidal acvity [26]
Atomoxene (brand
name Straera)
A non-smulant drug in the treatment of
aenon-decit hyperacvity disorder and a
selecve noradrenaline reuptake inhibitor [27]
Octodrine
To treat Bronchis, Laryngis, [28] an-fun-
gal,[29] an-microbial, [30] an-tumor acvies
[28]
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Library/ID – (Plan-
tago Lanceolata)Biological Acvity Structure Library/ID –
(Plantago Major)Biological Acvity Structure
Benzyl alcohol,
p-hydroxy-.alpha.-
[(methylamino)
methyl]- / Syneph-
rine
Synephrine is a primary synthesis drug devel-
oped as a sympathomimec agent with phar-
macological acvies, such as vasoconstricon,
blood pressure elevaon, and bronchial muscle
relaxaon [31].
Epinephrine To treat bronchiolis, [32]
and anaphylaxis [33]
Bicyclo[2.2.1]heptan-
2-one, 4,7,7-trimeth-
yl-, semicarbazone
An-candida, an-inammatory acvies [34] 3,4-Methylenedioxy-
amphetamine
An empathogen-entactogen, psychostimulant,
and psychedelic drug of the amphetamine family,
as a recreaonal drug [35]
(+)-Norpseudo-
ephedrine / Cathine
Cathine and norephedrine, phenylpropanol-
amines structurally related to amphetamine [36]
3-Methoxyamphet-
amine A designer drug alternave to MDMA [37]
Cyanoacetylurea
As a starng material for the synthesis of a
variety of heterocycles1 is easily prepared from
low-cost materials [38], a key intermediate in
the synthesis of 6-aminouracils, which possess
several biological acvies such as an-cancer
[39], an-viral [40], an-hypertensive [41],
inseccidal, herbicidal, acaricidal
acvies [42]
Metanephrine Inacve metabolite of epinephrine [43]
Desmethyldoxepin An-depressant properes [44] Mexilene An-arrhythmic acvity [45]
endo-Borneol An-bacterial, an-fungal acvies [46]
Benzenemetha-
nol, alpha.-(1-
aminoethyl)-2,5-
dimethoxy- /
Methoxamine
A blood-pressure increasing drug commonly used
for maintaining intraoperave hemodynamics
[47]
4-Fluorohistamine Substrate for several enzymes and inhibitor for
hisdine ammonia lyase [48] Acnobolin Anbioc, antumor, anbacterial [49]
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Library/ID – (Plan-
tago Lanceolata)Biological Acvity Structure Library/ID –
(Plantago Major)Biological Acvity Structure
Folic Acid Free radical scavenging, and an-oxidant acvi-
es [50] Nortriptyline Andepressant, as an analgesic in chronic back
pain [51]
Imidazole, 2-amino-
5-[(2-carboxy)vinyl]-
An-microbial, an-inammatory,[52] an-
cancer acvies [53]
1-[alpha-(1-Ada-
mantyl)benzylidene]
thiosemicarbazide
Thiosemicarbazone derivaves present a great
variety of biological acvies, such as an-viral,
an-cancer, an-tumor, an-inammatory, an-
amoebic, and an-microbial acvies [54].
N-2,4-Dnp-L-arginine An acvang eect on hepatocellular carcinoma
receptor B4 [55] Desmethyldoxepin
Desmethyldoxepin is the major acve metabo-
lite of doxepin (doxepin showed an-oxidant acvi-
es), [56] an an-depressant, and a drug metabolite
[57].
Northiaden A major acve metabolite of the tricyclic an-
depressant (TCA) dosulepin [58] 4-Fluorohistamine Substrate for several enzymes and inhibitor for
hisidine ammonia lyase [48]
Phenylpropanol-
amine
A decongestant, appete suppressant, [59-61]
cough, cold preparaons [62-63]
Cyclobutanol,
2-ethyl- Cyclobutanol as an an-microbial acvity [64]
Pterin-6-carboxylic
acid
An-cancer, an-viral [65] an-psychoc,
Moodstabilizer, an-parasite,[66] an-oxidant,
an-inammatory acvies [67]
Cyclopropyl carbinol
Biomedicine, avor, skin care and cosmec, skin-
care and cosmec, and bioenergy fungicides and
inseccides, [68] an intermediate used in chemical
laboratory research and development of organic
compounds and pharmaceucals [69]
Quinoline, 4-methyl-,
1-oxide An-cancer acvity [70] Cycloserine An anbioc used to treat tuberculosis [71]
Tocainide An-arrhythmic, local anesthecs,[72] an-
arrhythmic agent [73]
Methylpent-4-enyl-
amine
Flavor indicang volales characterized by ripening
[74]
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Colorectal cancer cells were incubated after treatment
with essential oils to udy the cytotoxic activities of P.
lanceolata and P. major. The essential oils of P. major
exhibited more antiproliferative properties on HCT-
116 at 72 h compared to P. lanceolata (IC50: 102.66 µg/
mL). IC50 values showed that P. major essential oil had
a greater cytotoxic eect on HCT-116 than HEK-293;
however, P. lanceolata showed almo the same eect on
cancer and normal cells (Table 5). The results indicated
that a very low IC50 of 5-FU (4.136 µg/mL) was required
to inhibit HCT-116 cell viability compared to the essen-
tial oil of P. lanceolata and P. major.
Toxicity assay on artemia salina
The general toxicity of the essential oils was assessed
again A. salina. At 25-100 µg/mL of the essential
oils, all of the nauplii were alive, indicating no toxicity
(LC50:2242.57 µg/mL and 1783.7 µg/mL) (Table 5). At
400 µg/mL of P. lanceolata and P. major, the percentage
of lethality was 8% and 12%, respectively. Although, the
K2Cr2O7 has shown to have a toxic eect (LC50 of 58.22
μg/mL).
Table 5. IC50 values of colorectal cancer cells and embryonic kidney normal cells and LC50 values of artemia salina by plantago
lanceolata and plantago major essential oils
Essenal Oils /Cell
HCT-116 (µg/mL) HEK-293 (µg/mL) Artemia Salina (µg/mL)
Mean±SD
24 h 48 h 72 h 24 h 48 h 72 h 24 h
Plantago lanceolata 622.54d±13.0 322.5b±17.5 158.33ab±12.9 508.65b±1.3 280.5ab±2.2 152.45ab±1.5 2242.57b±8.7
Plantago major 458.62a±8.5 262.45a±10.1 102.66a±9.3 566.82c±2.5 245.32a±7.0 224.45b±13.7 1783.7a±15.3
Notes: The analysis was performed separately every time. IC50 and LC50 values are the mean of the 3 replications±standard
deviation at 24, 48, and 72 h. The Duncan test was used for mean comparison (P<0.05). Charts with the same letters are not
statistically signicant. Values were calculated for 5-uorouracil (IC50:4.136 µg/mL) and Potassium dichromate (LC50:58.22
µg/mL) as positive controls.
Figure 1. Chromatogram of essential oils of the aerial part of plantago species
(A) Plantago Lanceolata and (B) Plantago Major
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Discussion
The presence of valuable compounds in P. lanceolata
can be a putative candidate for its application in mod-
ern medicine, as it has been used in traditional medicine
for many years. The following compounds were pres-
ent in this species: the anti-cancer compounds reported
in Table 4, such as linalool [11]; cyanoacetylurea [39];
imidazole, 2-amino-5-[(2-carboxy)vinyl]- [53]; pterin-
6-carboxylic acid [65]; quinoline, 4-methyl-, 1-oxide
[70]; anti-microbial compounds, including 1-octen-3-ol
[13]; 2-isopropoxyethylamine [19]; arginine [25]; endo-
borneol [46]; and imidazole, 2-amino-5-[(2-carboxy)
vinyl]- [52]. The anti-viral compounds, including 2-fu-
ranmethanol, 5-ethenyltetrahydro-.α., .α.,5-trimethyl-,
cis [16]; cyanoacetylurea [40]; pterin-6-carboxylic
acid [65]; anti-oxidant compounds, such as 2-furan-
methanol, 5-ethenyltetrahydro-.α., .α.,5-trimethyl-, cis
[16]; folic Acid [50]; pterin-6-carboxylic acid [67];
anti-inammatory, such as linalool [11], imidazole,
2-amino-5-[(2-carboxy)vinyl]- [52]; bicyclo[2.2.1]
heptan-2-one, 4,7,7-trimethyl-, semicarbazone [34];
pterin-6-carboxylic acid [67]. Meanwhile, the anti-
malaria compound 8-amino-6-methoxyquinoline [21]
was found in the analysis of P. lanceolata essential oil.
It was revealed that the common components of essen-
tial oil are fatty acids [75]. For inance, Fons reported
palmitic acid in the essential oil of P. lanceolata leaves
[76]. Bajer et al. used GC/MS and GC/FID techniques
to udy the qualitative and semi-quantitative content of
Figure 2. Common volatile composition of plantago lanceolata and plantago major
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July 2022. Volume 8. Number 3
volatile conituents in the essential oil, respectively. In
their udy, the main aroma conituents of P. lanceolata
leaves were groups of fatty acids 28.0% – 52.1% (the
mo abundant palmitic acid 15.3% –32.0%), oxidated
monoterpenes 4.3% – 13.2% with linalool 2.7% – 3.5%,
ketones and aldehydes 6.9%–10.0% with pentyl vinyl
ketone 2.0% –3.4%, and alcohols 3.8%–9.2% with
1-octen-3-ol 2.4%–8.2%. They pointed out that apoca-
rotenoids (1.5%–2.3%) are the important conituents
because of their intense fragrance and they were identi-
ed in a relatively high amount. The importance is in its
potential manufacture control of raw material to supply
food supplements [1]. The high content of 1-octen-3-ol
(up to 8.2%) has been observed in the Bajer et al., 2016
udy [1] in accordance with Fons [76]. This compound
in the present udy was about 1.27%.
Other udies showed that P. major essential oil has
anti-tumor and anti-cancer activities because octodrine
[28] and 1-[α-(1-adamantyl) benzylidene] thiosemicar-
bazide [54] were present in P. major essential oil. The an-
ti-microbial components, i.e., 2-dodecen-1-yl(-) succinic
anhydride [12]; 2-chloroacetamide [17]; isoborneol [23];
octodrine [30]; actinobolin [49]; 1-[α-(1-adamantyl)
benzylidene] thiosemicarbazide [54]; cyclobutanol,
2-ethyl- [64]; antiviral compounds, including isobor-
neol [22]; 1-[α-(1-adamantyl) benzylidene] thiosemi-
carbazide [54]; antioxidant and anti-inammatory com-
pounds, such as 2-dodecen-1-yl(-)succinic anhydride
[12]; desmethyldoxepin [56] and 1-[α-(1-adamantyl)
benzylidene] thiosemicarbazide [54] were observed in
the analysis of P. major essential oil. Some of the com-
pounds identied in the analysis of the P. major essential
oil showed important characteriics, such as cycloser-
ine [71] and actinobolin [49] which are antibiotic drugs
(0.75% and 0.13%) and isoborneol is anti-infective
(1.17%) [22] (Table 4). The percentage and dierences
in the amount of these compounds depend on many fac-
tors, such as climatic conditions, type of region, plant
growth conditions, and harveing methods.
The present udy indicated that a very low IC50 value
of 5-FU was required to inhibit HCT-116 cell viability
compared to the essential oil of P. lanceolata and P. ma-
jor. However, the IC50 obtained for the essential oil of
P.lanceolata and P.major were valuable and has increas-
ingly important medical applications. Our previous ud-
ies reported the cytotoxic eects of alcoholic and ace-
tonic extracts of P.major leaf and root on HCT-116 and
HEK-293. The P. major root extract was more eective
than the aerial parts, and IC50 values for ethanolic, meth-
anolic, and acetonic root extracts were 405.59, 470.16,
and 82.26 μg/mL, respectively on HCT-116 at 72 h [77].
In a udy by Velasco-Lezama (2006), the cytotoxic ac-
tivity of P. major methanolic extract has been reported
on HCT-15 [78].
For the lethality of nauplii, if LC50, detected for each
sample, is more than 1000 µg/mL, it will be non-toxic
[79]. At 400 µg/mL of P. lanceolata and P. major, the
percentage of the lethality of nauplii was 8% and 12%,
respectively. Thus, the essential oils were not toxic.
Other researchers have also evaluated the toxicity ef-
fect of P. major methanolic extract on A. salina and A.
uramiana with LC50 of 303.7 μg/mL [80]. The LC50
values of Plantago squarrosa Murray extracts were more
than 1000 μg/mL; therefore, the extracts were non-toxic
in the Artemia franciscana bioassay [81]. Our previous
udy showed that at all concentrations of ethanolic ex-
tracts of P.major aerial parts and roots, no toxicity was
observed [77].
Conclusions
Given the non-aromatic nature of P. lanceolata and
P. major and the very small amount of essential oil in
these plants, mo phytochemical udies are usually
performed on their extracts. Therefore, in the present
udy, the essential oils analysis of two well-known spe-
cies of Plantago was conducted to discover the valuable
compositions. The hydrodiillation method enabled us
to gain a great number of volatile conituents, which is
evident from the number of peaks that occurred in chro-
matograms. The mo abundant family of compounds
was amines. There were also identied acids, alcohols,
alkaloids, alkanes, alkenes, amides, amino acids, eers,
ketones, phenols, and terpenes that mo of the terpenes
were oxidated as monoterpenes. On the other hand, ni-
triles, oximes, and organic compounds were found in a
relatively small amount.
Regarding the chemical compounds identied in the P.
lanceolata and P. major essential oils, these components
could be employed as an important economical source in
the pharmaceutical and chemical induries. We intend
to udy their biological activities in the future.
Ethical Considerations
Compliance with ethical guidelines
There were no ethical considerations to be considered in
this research.
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
220
July 2022. Volume 8. Number 3
Funding
The paper was extracted from the PhD. Dissertation of
the r author at Department of Plant production and
Genetics, Faculty of Agriculture, Zanjan University of
Medical Sciences (Grant number: A-12-848-35).
Authors' contributions
Project adminiration, inveigation, formal analy-
sis, and writing-original draft: Samaneh Rahamouz-
Haghighi; Formal analysis, methodology, and valida-
tion: Alireza Yazdinezhad; Funding and supervision:
Khadijeh Bagheri; Funding, supervision, conceptual-
ization, and editing of the English version of the manu-
script: Ali Shara.
Conict of interest
The authors declare that there are no conicts of inter-
e regarding the publication of this article.
Acknowledgments
This work was supported by Zanjan Pharmaceutical
Biotechnology Research Center, Zanjan University of
Medical Sciences, Zanjan, Iran (Grant number: A-12-
848-35). In addition, the authors would like to thank the
authority of the School of Pharmacy, Zanjan University
of Medical Sciences, Zanjan, Iran.
References
[1] Bajer T, Janda V, Bajerová P, Kremr D, Eisner A, Ventura K.
Chemical composition of essential oils from Plantago lanceo-
lata L. leaves extracted by hydrodistillation. J Food Sci Tech-
nol 2016; 53(3):1576-84. [DOI:10.1007/s13197-015-2083-x]
[PMID] [PMCID]
[2] Ji X, Hou C, Guo X. Physicochemical properties, structures,
bioactivities and future prospective for polysaccharides
from Plantago L.(Plantaginaceae): A review. Int J Biol Macro-
mol. 2019; 135:637-46. [DOI:10.1016/j.ijbiomac.2019.05.211]
[PMID]
[3] Najaan Y, Hamedi SS, Farshchi MK, Feyzabadi Z. Plantago
major in Traditional Persian Medicine and modern phy-
totherapy: A narrative review. Electron Physician. 2018;
10(2):6390-9. [PMID] [PMCID]
[4] Gálvez M, Martín-Cordero C, Houghton PJ, Ayuso MJ. An-
tioxidant activity of methanol extracts obtained from Plan-
tago species. J Agric Food Chem. 2005; 53(6):1927-33. [PMID]
[5] Samuelsen AB. The traditional uses, chemical constituents
and biological activities of Plantago major L. A review. J
Ethnopharmacol. 2000; 71(1-2):1-21. [DOI:10.1016/S0378-
8741(00)00212-9]
[6] Weryszko-Chmielewska E, Matysik-Wozniak A, Sulborska
A, Rejdak R. Commercially important properties of plants
of the genus Plantago. Acta Agrobot. 2012; 65(1):11-20.
[DOI:10.5586/aa.2012.038]
[7] Akbari J, Saeedi M, Morteza-Semnani K, Zarrabi B, Ros-
tamkalaei SS, Kelidari HR. The effect of Plantago major seed
mucilage combined with carbopol on the release prole and
bioadhesive properties of propranolol HCl buccoadhesive
tablets. Pharm Biomed Res. 2016; 2(2):84-100. [DOI:10.18869/
acadpub.pbr.2.2.84]
[8] Khalaf HAA, Mahdi MF, Abaas IS. Preliminary phytochemi-
cal and GC-MS analysis of chemical constituents of Iraqi
Plantago lanceoleta L. Al-Mustansiriyah J Pharm Sci. 2018;
18(2):114-21. [DOI:10.32947/ajps.18.02.0381]
[9] Plumb JA. Cell sensitivity assays: Clonogenic assay. Meth-
ods Mol Med. 2004; 88:159-64. [PMID]
[10] Rajabi S, Ramazani A, Hamidi M, Naji T. Artemia salina as
a model organism in toxicity assessment of nanoparticles.
Daru. 2015; 23(1):20. [PMID] [PMCID]
[11] Al-Marzoqi AH, Hameed IH, Idan SA. Analysis of bioac-
tive chemical components of two medicinal plants (Corian-
drum sativum and Melia azedarach) leaves using gas chro-
matography-mass spectrometry (GC-MS). Afr J Biotechnol.
2015; 14(40):2812-30. [DOI:10.5897/AJB2015.14956]
[12] Jatin RR, Priya R S. Determination of bioactive components
of cynodon dactylon by GC-MS analysis & it’s in vitro an-
timicrobial activity. Int J Pharm Life Sci. 2016; 7(1):4880-5.
[Link]
[13] Xiong C, Li Q, Li S, Chen C, Chen Z, Huang W. In vitro an-
timicrobial activities and mechanism of 1-octen-3-ol against
food-related bacteria and pathogenic fungi. J Oleo Sci. 2017;
66(9):1041-9. [PMID]
[14] Chitarra GS, Abee T, Rombouts FM, Posthumus MA, Di-
jksterhuis J. Germination of penicillium paneum conidia is
regulated by 1-octen-3-ol, a volatile self-inhibitor. Appl En-
viron Microbiol. 2004; 70(5):2823-9. [PMID]
[15] Zin T, Sundaram CS, Rao UM. In silico analysis of the phe-
nylephrine from the sand crab (emerita asiatica) for its an-
timicrobial activities. Res J Pharm Technol. 2019; 12(5):01-4.
[DOI:10.5958/0974-360X.2019.00356.1]
[16] Hameed IH, Altameme HJ, Idan SA. Artemisia annua: Bio-
chemical products analysis of methanolic aerial parts extract
and anti-microbial capacity. Res J Pharm Biol Chem Sci.
2016; 7(2):1843-68. [Link]
[17] Dabbagh A, Tajbakhsh A, Talebi Z, Rajaei S. Cardiovas-
cular pharmacology in adult patients undergoing cardiac
surgery. In: Dabbagh A, Esmailian F, Aranki S. Postopera-
tive critical care for adult cardiac surgical patients. Cham:
Springer; 2018. [DOI:10.1007/978-3-319-75747-6_4]
[18] Katke SA, Amrutkar SV, Bhor RJ, Khairnar MV. Synthesis
of biologically active 2-chloro-N-alkyl/aryl acetamide de-
rivatives. Int J Pharm Sci Res. 2011; 2(7):148-56. [Link]
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
221
July 2022. Volume 8. Number 3
[19] Devi RB, Barkath T, Vijayaraghavan P, Rejiniemon TS. GC-
MS analysis of phytochemical from Psidium guajava Linn
leaf extract and their in-vitro antimicrobial activities. Int J
Pharma Bio Sci. 2019; 8:583-9. [Link]
[20] Sisler EC, Blankenship SM, Guest M. Competition of cy-
clooctenes and cyclooctadienes for ethylene binding and
activity in plants. Plant Growth Regul. 1990; 9:157-64.
[DOI:10.1007/BF00027443]
[21] Martinez ML, Campagna MN, Ratti MS, Nocito I, Serra
E, Gattuso S, et al. Trypanocide activity of Castela coccinea
Griseb. extracts. Boletín Latinoamericano y del Caribe de
Plantas Medicinales y Aromáticas. 2009; 8(3):211-8. [Link]
[22] Armaka M, Papanikolaou E, Sivropoulou A, Arsenakis
M. Antiviral properties of isoborneol, a potent inhibitor of
Herpes simplex virus type 1. Antivir Res. 1999; 43(2):79-92.
[DOI:10.1016/S0166-3542(99)00036-4]
[23] Tabanca N, Kırımer N, Demirci B, Demirci F, Başer KH.
Composition and antimicrobial activity of the essential oils
of Micromeria cristata subsp. phrygia and the enantiomeric
distribution of borneol. J Agric Food Chem. 2001; 49(9):4300-
3. [PMID]
[24] Asressu KH, Tesema TK. Chemical and antimicrobial in-
vestigations on essential oil of Rosmarinus ofcinalis leaves
grown in Ethiopia and comparison with other countries. J
Appl Pharm Sci. 2014; 6:132-42. [DOI:10.21065/19204159.6.
3.112]
[25] Wessolowski A, Bienert M, Dathe M. Antimicrobial activity
of arginine‐and tryptophan‐rich hexapeptides: The effects of
aromatic clusters, d‐amino acid substitution and cyclization.
J Pept Res. 2004; 64(4):159-69. [PMID]
[26] Kenaga EE, Allison WE. Commercial and experimental or-
ganic insecticides (1969 revision). Bulletin Entomol Soc Am.
1969; 15(2):85-148. [DOI:10.1093/besa/15.2.85]
[27] Dean L. Atomoxetine therapy and CYP2D6 genotype. In:
Pratt VM, Scott SA, Pirmohamed M, Esquivel B, Kane MS,
Kattman BL,et al, editors. Medical genetics summaries.
Bethesda: National Center for Biotechnology Information
(US); 2020. [PMID]
[28] Catalani V, Prilutskaya M, Al-Imam A, Marrinan S, Elghar-
ably Y, Zloh M, et al. Octodrine: New questions and chal-
lenges in sport supplements. Brain sci. 2018; 8(2):34. [PMID]
[29] Kim K, Zilbermintz L, Martchenko M. Repurposing FDA
approved drugs against the human fungal pathogen, Can-
dida albicans. Ann Clin Microbiol J Antimicrob. 2015; 14:32.
[PMID] [PMCID]
[30] Niu H, Cui P, Shi W, Zhang S, Feng J, Wang Y, et al. Iden-
tication of anti-persister activity against uropathogenic Es-
cherichia coli from a clinical drug library. Antibiotics Basel.
2015; 4(2):179-87. [PMID] [PMCID]
[31] Pellati F, Benvenuti S, Melegari M. Enantioselective LC
analysis of synephrine in natural products on a protein-
based chiral stationary phase. J Pharm Biomed Anal. 2005;
37(5):839-49. [PMID]
[32] Hartling L, Bialy LM, Vandermeer B, Tjosvold L, Johnson
DW, Plint AC, et al. Epinephrine for bronchiolitis. Cochrane
Database Syst Rev. 2011; 6:1-138. [DOI:10.1002/14651858.
CD003123.pub3]
[33] Parish HG, Morton JR, Brown JC. A systematic review of
epinephrine stability and sterility with storage in a syringe.
Allergy Asthma Clin Immunol. 2019; 15:7. [PMID]
[34] Kamal SA, Hamza LF, Ibraheam IA. Characterization of
antifungal metabolites produced by Aeromonas hydrophila
and analysis of its chemical compounds using GC-MS. Res
J Pharm Technol. 2017; 10(11):3845-51. [DOI:10.5958/0974-
360X.2017.00697.7]
[35] Crean RD, Davis SA, Von Huben SN, Lay CC, Katner SN,
Taffe MA. Effects of (±) 3, 4-methylenedioxymethampheta-
mine,(±) 3, 4-methylenedioxyamphetamine and metham-
phetamine on temperature and activity in Rhesus macaques.
Neuroscience. 2006; 142(2):515-25. [PMID] [PMCID]
[36] Adeoya-Osiguwa S, Fraser LR. Cathine, an ampheta-
mine-related compound, acts on mammalian spermato-
zoa via β1-and α2A-adrenergic receptors in a capacitation
state-dependent manner. Hum Reprod. 2007; 22(3):756-65.
[DOI:10.1093/humrep/del454] [PMID]
[37] Dal Cason TA. A re-examination of the mono-methoxy po-
sitional ring isomers of amphetamine, methamphetamine
and phenyl-2-propanone. Forensic Sci Int. 2001; 119(2):168-
94. [DOI:10.1016/S0379-0738(00)00425-4]
[38] Allam YA, Swellem RH, Nawwar GA. Cyanoacetylurea
in heterocyclic synthesis: A simple synthesis of heterocyclic
condensed uracils. J Chem Res. 2001; 2001(8):346-8. [DOI:10.
3184/030823401103170034]
[39] Ohwada S, Ikeya T, Yokomori T, Kusaba T, Roppongi T,
Takahashi T, et al. Adjuvant immunochemotherapy with
oral Tegafur/Uracil plus PSK in patients with stage II or III
colorectal cancer: A randomised controlled study. Br J can-
cer. 2004; 90(5):1003-10. [PMID] [PMCID]
[40] Peters D, Hörnfeldt AB, Gronowitz S, Johansson NG. Syn-
thesis and antiviral activity of various 5-substituted 2′-de-
oxyuridines and-cytidines. Nucleosides Nucleotides.1992;
11(6):1151-73. [DOI:10.1080/07328319208018333]
[41] Dooley M, Goa KL.Urapidil. A reappraisal of its use in the man-
agement of hypertension. Drugs. 1998; 56(5):929-55. [PMID]
[42] Yagi K, Akimoto K, Mimori N, Miyake T, Kudo M, Arai K,
et al. Synthesis and insecticidal/acaricidal activity of novel
3‐(2, 4, 6‐trisubstituted phenyl) uracil derivatives. Pest Man-
ag Sci. (Formerly Pestic Sci). 2000; 56(1):65-73. [DOI:10.1002/
(SICI)1526-4998(200001)56:13.0.CO;2-S]
[43] Lenders JW, Pacak K, Walther MM, Linehan WM, Mannelli
M, Friberg P, et al. Biochemical diagnosis of pheochromocy-
toma: Which test is best? JAMA. 2002; 287(11):1427-34. [PMID]
[44] Badenhorst D, Sutherland F, De Jager A, Scanes T, Hundt
H, Swart K, et al. Determination of doxepin and desmethyl-
doxepin in human plasma using liquid chromatography-
tandem mass spectrometry. J Chromatogr B: Biome Sci Appl.
2000; 742(1):91-8. [DOI:10.1016/S0378-4347(00)00136-5]
[45] Canavero S. Central pain syndrome: Pathophysiology, di-
agnosis and management. Cambridge: Cambridge Univer-
sity Press; 2011. [DOI:10.1017/CBO9780511845673]
[46] Ashraf SA, Al-Shammari E, Hussain T, Tajuddin S, Pan-
da BP. In-vitro antimicrobial activity and identication of
bioactive components using GC-MS of commercially avail-
able essential oils in Saudi Arabia. J Food Sci Technol. 2017;
54(12):3948-58. [PMID]
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
222
July 2022. Volume 8. Number 3
[47] Sun S, Sun D, Yang L, Han J, Liu R, Wang L. Dose-depend-
ent effects of intravenous methoxamine infusion during
hip-joint replacement surgery on postoperative cognitive
dysfunction and blood TNF-α level in elderly patients:
A randomized controlled trial. BMC Anesthesiol. 2017;
17(1):75. [PMID]
[48] Sundaram S, Prabhakaran J. Effect of 1-methylcyclopro-
pene (1-MCP) on volatile compound production in papaya
(Carica papaya L.) fruit. Pharm Innovat J. 2017; 6:532-6.
[Link]
[49] Imuta S, Tanimoto H, Momose MK, Chida N. Total synthe-
sis of actinobolin from d-glucose by way of the stereoselec-
tive three-component coupling reaction. Tetrahedron. 2006;
62(29):6926-44. [DOI:10.1016/j.tet.2006.04.079]
[50] Joshi R, Adhikari S, Patro BS, Chattopadhyay S, Mukherjee
T. Free radical scavenging behavior of folic acid: Evidence
for possible antioxidant activity. Free Radic Biol Med. 2001;
30(12):1390-9. [DOI:10.1016/S0891-5849(01)00543-3]
[51] Atkinson HJ, Slater MA, Williams RA, Zisook S, Patterson
TL, Grant I, et al. A placebo-controlled randomized clinical
trial of nortriptyline for chronic low back pain. Pain. 1998;
76(3):287-96. [DOI:10.1016/S0304-3959(98)00064-5]
[52] Kadhim MJ, Mohammed GJ, Hameed IH. In vitro antibac-
terial, antifungal and phytochemical analysis of methanolic
extract of fruit Cassia stula. Orient J Chem. 2016; 32(3):1329-
46. [DOI:10.13005/ojc/320307]
[53] Gajjala K, Anisetti R, Dodla JP, Rathod AK. Phytochemi-
cal investigation and cytotoxic activity of hydro alcoholic
fraction of Trianthema decandra. Indian J Biotechnol. 2019;
18:193-203. https://scholar.google.com/scholar?hl=en&as_
sdt=0%2C5&q
[54] Sardari S, Feizi S, Rezayan AH, Azerang P, Mohammad
Shahcheragh S, Ghavami G, et al. Synthesis and biological
evaluation of thiosemicarbazide derivatives endowed with
high activity toward Mycobacterium bovis. Iran J Pharm
Res. 2017; 16(3):1128-40. [DOI:10.22037/IJPR.2017.2069]
[55] Kamstra RL, Freywald A, Floriano WB. N‐(2, 4)‐dinitro-
phenyl‐L‐arginine interacts with EphB4 and functions as
an EphB4 Kinase modulator. Chem Biol Drug Des. 2015;
86(4):476-86. [PMID]
[56] Palchoudhuri S, Mukhopadhyay D, Roy DS, Ghosh B, Das
S, Dastidar SG. The antidepressant drug doxepin: A promis-
ing antioxidant. Asian J Pharm Clin Res. 2017; 10(3):97-102.
[DOI:10.22159/ajpcr.2017.v10i3.15149]
[57] Kirchheiner J, Henckel HB, Franke L, Meineke I, Tzvetkov
M, Uebelhack R, et al. Impact of the CYP2D6 ultra-rapid me-
tabolizer genotype on doxepin pharmacokinetics and sero-
tonin in platelets. Pharmacogenet Genomics. 2005; 15(8):579-
87. [PMID]
[58] Williams DA. Lemke TL. Foye’s principles of medicinal
chemistry. Philadelphia: Lippincott Williams & Wilkins;
2012. [Link]
[59] Swiss Pharmaceutical Society. Index nominum 2000: In-
ternational drug directory. Guildford: Medpharm Scientic
Publishers; 2000. [Link]
[60] Elks J. The dictionary of drugs: Chemical data: Chemical
data, structures and bibliographies. New York: Springer;
2014. [Link]
[61] Morton IK, Hall JM. Concise dictionary of pharmacological
agents: Properties and synonyms. Berlin: Springer Science &
Business Media; 1999. [DOI:10.1007/978-94-011-4439-1]
[62] Gupta RC. Veterinary toxicology: Basic and clinical princi-
ples. Amsterdam: Elsevier Science; 2012. [Link]
[63] Adams HR, Papich MG, Riviere JE. Veterinary pharmacol-
ogy and therapeutics, 9th ed. Ames, Iowa: Wiley-Blackwell,
2009. https://lib.ugent.be/catalog/rug01:001318502
[64] Ara I, Shinwari M, Rashed S, Bakir M. Evaluation of an-
timicrobial properties of two different extracts of Juglans
regia tree bark and search for their compounds using gas
chromatography-mass spectrum. Int J Biol. 2013; 5(2):92-102.
[DOI:10.5539/ijb.v5n2p92]
[65] Al-Rubaye AF, Kaizal AF, Hameed IH. Phytochemical
screening of methanolic leaves extract of Malva sylves-
tris. Int J Pharmacog Phytochem Res. 2017; 9(4):537-52.
[DOI:10.25258/phyto.v9i4.8127]
[66] Hussein AO, Mohammed GJ, Hadi MY, Hameed IH. Phy-
tochemical screening of methanolic dried galls extract of
Quercus infectoria using gas chromatography-mass spec-
trometry (GC-MS) and Fourier transform-infrared (FT-IR).
J Pharmacogn Phytotherapy. 2016; 8:49-59. [DOI:10.5897/
JPP2015.0368]
[67] Kadhim MJ. In vitro antifungal potential of Acinetobacter
baumannii and determination of its chemical composition
by gas chromatography-mass spectrometry. Der Pharma
Chemica. 2016; 8(19):657-65. [Link]
[68] Perumal R, Albertmanoharan S, Pemiah B. GC-MS evi-
dence based herbocure from indian system of medicine
for stomach disorders in vets. Asian J Anim Vet Adv. 2018;
13(1): 73-84. [DOI:10.3923/ajava.2018.73.84]
[69] Suo C, Sun Z, Wang Y, Dong S, Lv C, Wang T, et al. Ac-
tive components in leaves of Rhus chinensis Mill. Therm Sci.
2020; 24(3A):1729-35. [DOI:10.2298/TSCI190529045S]
[70] Jain S, Chandra V, Jain PK, Pathak K, Pathak D, Vaidya A.
Comprehensive review on current developments of quino-
line-based anticancer agents. Arab J Chem. 2019; 12(8):4920-
46. [DOI:10.1016/j.arabjc.2016.10.009]
[71] Hwang TJ, Wares DF, Jafarov A, Jakubowiak W, Nunn
P, Keshavjee S. Safety of cycloserine and terizidone for the
treatment of drug-resistant tuberculosis: A meta-analysis.
Int J Tuberc Lung Dis. 2013; 17(10):1257-66. [PMID]
[72] Nguyen LA, He H, Pham-Huy C. Chiral drugs: An over-
view. Int J Biomed Sci. 2006; 2(2):85-100. [PMID]
[73] Ayodele OO, Onajobi FD, Osoniyi OR. Phytochemical pro-
ling of the hexane fraction of crassocephalum crepidioides
benth S. Moore leaves by GC-MS. Afr J Pure Appl Chem.
2020; 14(1):1-8. [DOI:10.5897/AJPAC2019.0815]
[74] Malekzadeh H, Fatemi MH. Analysis of avor volatiles of
some Iranian rice cultivars by optimized static headspace
gas chromatography-mass spectrometry. J Iran Chem Soc.
2015; 12:2245-51. [DOI:10.1007/s13738-015-0703-z]
[75] Clarke S. Essential chemistry for aromatherapyk. Church-
illLivingstone, London: Elsevier Health Sciences; 2009.
https://www.sciencedirect.com/book/9780443104039/
essential-chemistry-for-aromatherapy#book-info
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
223
July 2022. Volume 8. Number 3
[76] Fons F, Rapior S, Gargadennec A, Andary C, Bessière JM.
Volatile components of Plantago lanceolata (Plantaginaceae).
Acta Bot. Gall. 1998; 145(4):265-9. [DOI:10.1080/12538078.1
998.10516306]
[77] Rahamooz-Haghighi S, Bagheri K, Danafar H, Shara A.
Anti-proliferative properties, biocompatibility, and chemi-
cal composition of different extracts of plantago major medici-
nal plant. Iran Biomed J. 2021; 25(2):106-16. [PMID]
[78] Velasco-Lezama R, Tapia-Aguilar R, Román-Ramos R, Ve-
ga-Avila E, Pérez-Gutiérrez MS. Effect of Plantago major on
cell proliferation in vitro. J Ethnopharmacol. 2006; 103(1):36-
42. [PMID]
[79] Ruebhart DR, Wickramasinghe W, Cock IE. Protective
efcacy of the antioxidants vitamin E and Trolox against
Microcystis aeruginosa and microcystin-LR in Artemia
franciscana nauplii. J Toxicol Environ Health Part A. 2009;
72(24):1567-75. [PMID]
[80] Mirzaei M, Mirzaei A. Comparison of the Artemia salina and
Artemia uramiana bioassays for toxicity of 4 Iranian medici-
nal plants. Int J Biol Sci. 2013; 2(3):49-54. [Link]
[81] Omer E, Elshamy AI, Nassar M, Shalom J, White A, Cock
IE. Plantago squarrosa Murray extracts inhibit the growth
of some bacterial triggers of autoimmune diseases: GC-MS
analysis of an inhibitory extract. Inammopharmacology
2019; 27(2):373-85. [PMID]
Rahamouz Haghighi S, et al. Volatile Oils and Toxicity of P.lanceolata and P. major Essential Oils. PBR. 2022; 8(3)::205-224
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