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A Review on Traditional Uses, Phytochemistry and Pharmacological Activities of Calendula officinalis Linn

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Natural Product Communications
Authors:
  • Madan Bhandari Academy of Health Sciences

Abstract and Figures

Calendula officinalis Linn, known as pot marigold, is a plant that belongs to the Asteraceae family. Various online bibliographic databases namely, Google scholar, PubMed, SciFinder, and Web of Science were used for integrating information. Calendula officinalis is extensively used in Homoeopathic, Unani, and Ayurvedic system of medication as diaphoretic, analgesic, antiseptic, and anti-inflammatory agents and used to treat gynaecological issues, gastro-intestinal disorders, inflammations of oral and pharyangeal mucosa, eye problems, skin injuries and certain burns, poor eyesight, and menstrual irregularities. Several studies have shown that Calendula officinalis is a major source of diverse classes of bioactive compounds namely terpenoids, flavonoids, coumarines, quinones, and carotenoids. Various in vivo and in vitro assessment of Calendula officinalis's pharmacological activity suggest that the plant has antidiabetic, anti-inflammatory, anti-tumor, anticancer, and gastroprotective activity. This review compiles the information about pharmacological activities, traditional medicinal uses, and phytochemicals present in Calendula officinalis.
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A Review on Traditional Uses, Phytochemistry
and Pharmacological Activities of
Calendula ofcinalis Linn
Biswash Sapkota
1
and Paridhi Kunwar
2
Abstract
Calendula ofcinalis Linn, known as pot marigold, is a plant that belongs to the Asteraceae family. Various online bibliographic data-
bases namely, Google scholar, PubMed, SciFinder, and Web of Science were used for integrating information. Calendula ofcinalis is
extensively used in Homoeopathic, Unani, and Ayurvedic system of medication as diaphoretic, analgesic, antiseptic, and anti-inam-
matory agents and used to treat gynaecological issues, gastro-intestinal disorders, inammations of oral and pharyangeal mucosa, eye
problems, skin injuries and certain burns, poor eyesight, and menstrual irregularities. Several studies have shown that Calendula of-
cinalis is a major source of diverse classes of bioactive compounds namely terpenoids, avonoids, coumarines, quinones, and carot-
enoids. Various in vivo and in vitro assessment of Calendula ofcinaliss pharmacological activity suggest that the plant has antidiabetic,
anti-inammatory, anti-tumor, anticancer, and gastroprotective activity. This review compiles the information about pharmacological
activities, traditional medicinal uses, and phytochemicals present in Calendula ofcinalis.
Keywords
Calendula ofcinalis, pharmacological uses, phytochemistry, traditional uses
Received: January 5th, 2024; Accepted: May 14th, 2024.
Introduction
The plant Calendula ofcinalis Linn (C. ofcinalis) commonly called
as pot marigold is a owering plants belonging to Asteraceae
family and Calenduleae tribe.
1-3
The name calendula comes
from the Latin word calendas,which means rst of the
month.
4
The term marigold comes from calendula, which
was formerly known as goldsand was connected to
Queen Mary and the Virgin Mary.
5
C. ofcinalis is sessile,
hispid, acute, oblanceolate, annual, or biennial herbaceous
plant that is between 30 and 60 centimeters height. The
leaves are hairy, alternating, petiolate, oblong, and spatulate,
and have edges that are either whole or have few teeth. The
lower leaves have rounded tips and oval shape while upper
leaves are lance-shaped with pointed tips. The leaves blade
length range from 2 to 4 inches.
6
It has multiple secondary
roots and possesses tap root with numerous secondary
roots.
7
Figure 1 shows various parts of C. ofcinalis which
includes ower, leaves and roots. The plant also contains essen-
tial oils which are produced from the owers and the glandular
hair present over the plant.
8
The daisy like owers are yellow to
bright orange in color, curved, sickle-shaped, tubular and ringed
achene, disc orets, and hermaphrodite.
9,10
The Calenduleae tribe has eight genera and more than 110
species, mostly found in South Africa.
11
The plant is indigenous
to Macaronesia East through Mediterranean region, Western
Asia, United States, Europe, Cyprus, Turkey, Iran, and few
other middle eastern countries as an ornamental plant.
12
The
larger scale cultivation is observed in India and China.
13-15
Many gardeners think calendula can readily adapt to any
climate and grow pleasantly since most soil types are good
for its growth. It prefers a pH range of 6.0 to 7.5, wet, light
sandy to clay, well-drained, poor to moderately rich soil, but
will tolerate calcareous soil. In subtropical summer, plants will
wither. Although seeds grow best in sunny spot and readily
sprout out with well-drained soil.
6,15,16
This study consolidates dispersed reports on the traditional
applications, pharmacological properties, and phytochemistry
of C. ofcinalis. However, our focus lies in highlighting the
importance of C.ofcinalis as a natural therapeutic remedy, bol-
stered by positive ndings in the literature. The aim of this
1
Department of Pharmacy and Clinical Pharmacology, Madan Bhandari
Academy of Health Sciences, Hetauda, Nepal
2
Department of Pharmacy, Crimson College, Butwal, Nepal
Corresponding Author:
Biswash Sapkota, Department of Pharmacy and Clinical Pharmacology, Madan
Bhandari Academy of Health Sciences, Hetauda, Nepal.
Email: biswash.sapkota@mbahs.edu.np
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provided the original work is attributed as specied on the SAGE and Open Access page (https://us.sagepub.com/en-us/nam/open-access-at-sage).
Review
Natural Product Communications
Volume 19(6): 122
© The Author(s) 2024
Article reuse guidelines:
sagepub.com/journals-permissions
DOI: 10.1177/1934578X241259021
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review is to succinctly outline existing research, bridge any
knowledge gaps, and present multiple avenues for researchers
already exploring the validation of traditional claims and the
safe, efcacious use of C. ofcinalis in treating diverse diseases.
Common Name
As indicated in Table 1, C. ofcinalis is frequently referred to by
various names in different languages worldwide.
Traditional Uses
Ayurveda, Unani, Homeopathic, and other traditional system of
medication utilize C. ofcinalis plant widely for curing different
ailments. C. ofcinalis is a diaphoretic, analgesic, antiseptic, and
anti-inammatory agents and used to treat gynaecological
issues, gastro-intestinal disorders, inammations of oral and
pharyangeal mucous, eye problems, skin injuries
20
and certain
burns,
21
poor eyesight, and menstrual irregularities.
8,22-24
It is
also used as sudoric, blood rener, and blood sugar
reducer.
25
The owers head has been utilized as tinctures,
balms, and salves for its cicatrizing, anti-tumor,
26
blood rener,
peptic-ulcer, astringent, diuretic, hypoglycemic, and antipyretic
properties.
27-30
In India, Calendula ointments are applied topically
to treat skin damage, gangrene, wounds,
31
acne, chicken pox,
mumps, scars, herpes, ulcers, frostbite, and wounds.
32
Insect
bites, carbuncles, varicosities, dermatitis, boils, and oral sores or
tooth ache can all be treated with its infusion.
33
For the treatment
of varicose veins, leaves are placed externally in fusion form.
Calendula drinks are used as gargles, eye washes and treatments
for inammatory disorders of the mucous membranes as well
as for diaper rashes, hemorrhoids, stomatitis, and conjunctivi-
tis.
34,35
Besides its use in human, it has been extensively used
in veterinary homeopathy as well.
36
Chemical Constituents
The entire C. ofcinalis plant is a rich source of many phytochem-
ical components, with varying amounts and qualities of these
components in different plant sections. The most prevalent
ones are terpenoids, avonoids, saponins, sterols, phenolic
acids, coumarins, quinines, amino acids, essential oil, and carot-
enoids as shown in Table 2.
34,37-40
Terpenoids
The therapeutic effects of the C. ofcinalis is caused by a signi-
cant group of secondary metabolites called terpenoids.
1,8,41
Various terpenoids isolated (Figure 2) are calenduladiol-3-
O-palmitate, stigmasterol, lupeol, Ψ-taraxasteol,
42
calenduladiol-
3-O-myristate, arnidiol-3-O-myristate, arnidiol-3-O-laurate,
arnidiol-3-O-palmitate, faradiol-3-O-laurate, faradiol-3-O-
palmitate, faradiol-3-O-myristate,
8,43
calendulosides A
H,
8,37,44
and calendulaglycoside A and B.
27,45
Flavonoids
Flavonoids are mostly present in owers of C. ofcinalis. Some of
the isolated avonoids (Figure 3)
14,37
are quercetin, calendoa-
side, calendoavoside, isoquercetin, calendoavobioside, rutin,
gallic acid,
46
isorhamnetin,
47
isorhamnetin-3-O-D-glycoside, nar-
cissin,
48
pinobanksin 3-acetate,
49
isorhamnetin-3-O-D-glycoside,
neohesperidoside, isorhamnetin-3-O-2G-rhamnosyl rutinoside,
quercetin-3-O-glucoside, isorhamnetin-3-O-2G quercetin, scopole-
tin-7-O-glucoside, isorhamnetin-3-O-glucoside calendoavobioside,
Figure 1. Different parts of C. ofcinalis.
Table 1. Common Name of CO in Various Languages.
S.N Languages Common name
1 English Pot marigold, Scoth Marigold, Calendula,
Marybud, Gold-bloom, Holligold, English
Marigold
2 Hindi Genda
3 French Souci des jardins
4 Spanish Maravilla, calendula
5 Portugese Wonders, boninas
6 French Calendula eur
7 Italian Calendula, orrancio
8 German Ringelblume
17-19
2Natural Product Communications
isorhamnetin-3-O-2G-rhamnosyl rutinoside, and isorhamne-
tin-3-O-2G.
50-54
Coumarins
The extract of C. ofcinalis contains (Figure 4)umbelliferone,
scopoletin, and esculetin.
55,56
The other coumarins glyscoside
isolated are isobaisseoside, haploperoside A, and haploperoside
D. A new coumarin glycoside, and neoisobaisseoside are
present on its owers.
57,58
Sterols
The leaves of C. ofcinalis are the major source of sterols.
Campesterol, sitosterol, cholestanol, campestanol, stigmastanol,
24-methylcholest-7-en-3β-ol, stigmast-7-en-3β-ol, cholesterol,
cholest-7-en-3-β-ol, umbelliferon, aesculetin,
36
24-methylcholesta-
5,22-dien-3β-ol, 24-methylenecholesterol, stigmasterol, and
clerosterol (Figure 5) were isolated from leaves.
59-62
Quinones
Phylloquinone, tocopherol, and plastoquinone are present in
chloroplast and ubiquinone, and phylloquinone (Figure 6) are
found in the leaves.
63
Essential Oil
The essential oils are predominately isolated from owers of C.
ofcinalis. The highest (0.97%) content of essential oils are
present during the full blooming stage while lowest (0.13%)
during pre-owering stage.
64
The essential oil contains
(Figure 7) calendic acid, 1,8-Cineol, p-cymene, transocimene,
thujene, α-pienene, sabinene, limonene, α-terpenene, Nonanal,
terpene-4-ol, 6-Methyl-5-heptene-2-one, 2-Pentylfuran, benze-
neacetaldehyde, γ-terpinene, 4-methylbenzaldehyde, terpinolene,
linalool, 2,6-dimethylcyclohexanol, cis-p-mentha-2-en-1-ol, epi-α-
muurolol, α-cadinol, terpinen-4-ol, α-terpineol, safranal,
n-decanal, β-cyclocitral, carvone, dihydrojasmone, p-thymol,
α-cubebene, α-Ionone, γ-muurolene, γ-cadinene, palmitic acid,
δ-cadinene, τ-muurolol,selina-6-en-4-ol, β-ionone, δ-cadinol,
τ-cadinol, β-oplopenone, α-cadinol, calamemene, viridiorol,
1-epi-cubenol, and cadalene.
65-69
Carotenoids
The yellow-orange hue of owersisattributedtothevast
amounts of carotenoids (Figure 8) found in C. ofcinalis inores-
cences; the exact shade of color is determined by the pigment
prole and content. In petal extracts of C. ofcinalis cultivars
with orange and yellow owers, 19 carotenoids were found.
Cultivars with orange owers were specicto10carotenoids.
67,70
Bakó et al examined carotenoids content and discovered
roughly eighteen distinct kinds of carotenoids: Neoxanthin, 9
´Z-auroxanthin, luteoxanthin, and neoxanthin (9
´Z)-violaxanthin, mutatoxanthin, αand β-carotene, avoxan-
thin, violaxanthin and 9´Z-lutein, astaxanthin, and 9/9
´Z-lutein (13/13´Z)-lutein.
71
Nineteen distinct carotenoids, including avoxanthin,
lutein-5,6-epoxide, (8´R)-luteoxanthin, and (8R/8´R)-lutein,
were discovered by Kishimoto et al (2005)auroxanthin,
lutein, antheraxanthin, (9Z)-lutein-5,6-epoxide, (9Z)-lutein, (5
Tab le 2. List of Phytochemicals Present in Calendula ofcinalis.
S.N
Chemical
constituents Isolated compound References
1. Terpenoids Faradiol-3-O-palmitate, faradiol-3-O-myristate, faradiol-3-O-laurate, arnidiol-3-O-palmitate,
arnidiol-3-O-myristate, arnidiol-3-O-laurate, calenduladiol-3-O-palmitate,
calenduladiol-3-O-myristate, calendulosides A-H
1,4,27-31
2 Flavonoids Quercetin, isorhamnetin, isoquercetin, isorhamnetin-3-O-D-glycoside, narcissin, calendoaside,
calendoavoside, rutin, isoquercetin neohesperidoside, isorhamnetin-3-O-2G-rhamnosyl rutinoside,
isorhamnetin-3-O-2G quercetin, isorhamnetin, isoquercetin, isorhamnetin-3-O-D-glycoside, narcissin,
calendoavoside, calendoavobioside,isorhamnetin-3-O-2G-rhamnosyl rutinoside,
isorhamnetin-3-O-2G
32-37
3 Coumarins Scopoletin, umbelliferone, esculetin
38-40
4 Quinones Plastoquinone, phylloquinone, tocopherol, ubiquinone
45,46
5 Carotenoids Neoxanthin, 9Z-neoxanthin, violaxanthin, luteoxanthin, auroxanthin, 9Z-violaxanthin, avoxanthin,
mutatoxanthin, 9Z-anthroxanthin, lutein, 9/9A-lutein, 13/13 Z-lutein, Z-cryptoxanthin, 13
Z-violaxanthin, antheraxanthin, mutatoxanthin epimer 1, mutatoxanthin epimer 2, lutein, 9/92-lutein,
cryptoxanthin, carotene
51,52
6 Amino acids Alanine, arginine, aspartic acid, asparagines, valine, histidine, glutamic acid, leucine, lysine, prolime, serine,
tyrosine, threonine, methionine, phenylalanine
53,54
7 Essential oils Thujene, pienene, sabinene,pienene, limonene, 1,8-cineol, p-cymene, trans-ocimene, terpenene, 3-carene,
nonanal, terpene-4-ol, 3-cylo, cadinene, cadinol, t-muurolol, limonene
47-50
8 Carbohydrates (1_3)-D-galactam, araban(1_3)-araban, alpha-L-rhamnan-(1_3)
55
9 Lipid Lauric, myristic, palmitic, stearic, oleic, linoleic and linolenic acid, D-(+)-9-hydroxy-10,12-ocatdecadienoic
acid
54,55
Sapkota and Kunwar 3
´Z/9´Z)-rubixanthin, α-, β-, and (5´Z)-carotene, δ-carotene,
(5Z/9Z/5´Z/9´Z)-lycopene, γ-carotene, and (5´Z)-γ-carotene,
(5Z/9Z/5Z)-lycopene, (5Z/9Z)-lycopene, and all-alycopene.
(5´Z)-γ-carotene and (5´Z/9´Z)-rubixanthin are two of their
carotenoid.
67,72
13Z-violaxanthin, lutein, 9/92-lutein, antherax-
anthin, and mutatoxanthin epimer 1 were all found in the
petals.
71
Amino Acids
It is stated that 15 amino acids are present in the ower extracts
in free form: leucine, valine, proline, histidine, asparagines, glutamic
acid, lysine, serine, threonine, methionine, alanine, arginine, aspartic
acid, tyrosine, and phenylalanine.
26,73,74
Carbohydrates
Carbohydrates like polysaccharides, I, II, and III are present.
75
There are also other monosaccharides such as glucose, xylose,
galactose, rhamnose, arabinose, and galacturonic acid.
76
Lipid and Fatty Acids
C. ofcinalis seeds contain neutral lipids, glycolipids, and phos-
pholipids among other lipids. Among the identied lipids in
C. ofcinalis are (Figure 9) palmitic acid, myristic acid, lauric acid,
stearic acid, limoleic acid, oleic acid, D-(+)-9-hydroxy-10,12-octa-
decadienoic acid, and linolenic acid.
73,77
The 11 genotypes of oils produced from calendula seeds
contained nineteen fatty acids. Calendic acid and linoleic acid
Figure 2. Chemical structure of terpenoids specic for C. ofcinalis.
14,37,38,44
. (continued)
4Natural Product Communications
Figure 2. Continued.
Sapkota and Kunwar 5
Figure 2. Continued.
6Natural Product Communications
Figure 3. Chemical structure of avonoids specic for C. ofcinalis.
14,37
. (continued)
Sapkota and Kunwar 7
(51.47%-57.63% and 28.5-31.9%) were discovered to be the
most prevalent fatty acids, followed by oleic acid (4.44-6.25%)
and palmitic acid (3.86-4.55%).
78,79
Pharmacological Activities
C. ofcinalis is frequently used in cure of numerous illnesses. It
may also be cytotoxic and stop the growth of tumors. It has
been used for medicinal properties as an antibacterial, antioxi-
dant, anti-inammatory, antiseptic, antiviral, hepatoprotective,
gastro-intestinal issues, obstetric issues, anti-mutagenic, and
antidiabetic as shown in Figure 10.
80-82
Antidiabetic Activity
Diabetes is a signicant global health concern due to its rapid
global expansion. Although diabetes mellitus (DM) is
common worldwide, more industrialized nations have higher
rates of the disease, particularly Type 2. DM ranks among the
top 10 causes of death worldwide, ranking alongside respiratory,
cardiovascular, and cancer disorders. According to reports,
people with diabetes between the ages of 20 and 99 caused
almost 5 million deaths globally in 2017.
83
Therefore, it has
always been desirable to identify new antidiabetic medications
composed of natural plants because some of their constituents
have been shown to be safe and effective alternatives to conven-
tional medications for DM.
84
Many researchers have reported
the antidiabetic activity of C. ofcinalis.
In vivo models using diabetic rats generated by streptozotocin
(STZ) and diabetic rats induced by alloxan were used to analyze
the antidiabetic activity of C. ofcinalis extracts. Additionally,
in vitro assays, such as amylase inhibitory activity and
α-glucosidase inhibition, were conducted to further evaluate
the extractspotential impact on diabetes. Chakraborthy et al
examined the antidiabetic properties of a hydroalcoholic
extract of C. ofcinalis in alloxan-induced diabetic rat models.
The extract, prepared with a 70:30 alcohol to water ratio
using the soxhlet extraction method, was orally administered
at 100 mg/kg body weight. This led to reductions in serum
lipid levels, urine sugar, and blood glucose in diabetic rats.
85
Additionally, in male rats with STZ-induced diabetes, treatment
with C. ofcinalis resulted in signicant increases in insulin, glu-
tathione, and serum inammatory cytokines (tumor necrosis
factor (TNF)-1 and interleukins-1 (IL-1)), alongside decreases
in malondialdehyde and glucose levels at doses of 200 and
400 mg/kg.
86
Moradkhani et al found a substantial reduction
in plasma glucose levels in STZ-induced diabetic rats treated
with a hydroalcoholic plant extract at 300 mg/kg.
87
In a
similar manner, the hydroalcoholic extract of owers (250
and 500 mg/kg) was assessed using a model of diabetic rats
Figure 3. Continued.
8Natural Product Communications
Figure 4. Chemical structure of coumarines specic for C. ofcinalis.
37
.
Sapkota and Kunwar 9
Figure 5. Chemical structure of sterols specic for C. ofcinalis.
57
.
10 Natural Product Communications
induced by STZ. The research revealed a noteworthy reduction
in fasting blood sugar (P< 0.05).
88
In another study, rats with
alloxan-induced diabetes had lower blood glucose levels when
exposed to a plant 100 mg/kg of hydroacholic extract.
85
Additionally, the plant extracts aid to lessen oxidative stress,
uctuating creatine kinase levels, weight loss, and other abnor-
malities in diabetic rats.
89
Neoisobaisseoside a glycoside was isolated from C. ofcinalis
and was found inhibiting α- amylase and α-glucosidase enzyme
signicantly which support the antidiabetic property of C. ofcina-
lis.
57
AC. ofcinalis leaf ethyl acetate and ethanolic extract-soluble
fraction demonstrated a noteworthy reduction in amylase activity.
Signicant and strong inhibitory effects on amylase were demon-
strated by isoquercitrin, 3,5-di-O-caffeoylquinic acid, isorhamne-
tin-3-O-β-D-glucopyranoside, and quercetin-3-O-(6′′-acetyl)-β-
D-glucopyranoside.
90
Thus, the presence of different avonoids,
glycosides, moderate amounts of alkaloids steroids and saponins
which possess higher antioxidant activity may be the reason for
exhibiting signicant antidiabetic activity.
27,91
Antioxidant Effects
The existence of free radicals, whether generated outside or
internally by the body, has been linked to diseases such as
liver cirrhosis, atherosclerosis, inammation, diabetes, neuro-
logical illnesses, and cancer. This connection between free
radicals and these diseases has spurred extensive research into
safe medications capable of neutralizing these radicals.
Numerous studies have highlighted the considerable antioxi-
dant capabilities of various plant extracts and plant-based prod-
ucts in this regard.
92,93
The 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scav-
enging, 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)
(ABTS) radical scavenging activity, nitric oxide (NO) radical
scavenging, ferric reducing antioxidant power (FRAP), superox-
ide radical scavenging, and in vivo procedures are the primary
methods used to assess the antioxidant activity of the C. ofcina-
lis plant.
C. ofcinalis ower extract efciently scavenged superoxide
radicals (inhibitory concentration 50 (IC50) =500 µg/mL),
hydroxyl radicals (IC50 =480 µg/mL), and lipid peroxidation
(IC50 =2000 µg/mL). It also demonstrated scavenging activity
against DPPH radicals (IC50 =100 µg/mL) and ABTS radicals
(IC50 =6.5 µg/mL). Moreover, the extract exhibited dose-
dependent scavenging of NO in culture (IC50 =575 µg/mL).
In vivo administration of oral calendula extract reduced macro-
phage superoxide production by 12.6% and 38.7% at dosages
of 100 and 250 mg/kg body weight, respectively. Additionally,
after 1 month of treatment, higher levels of glutathione and
catalase were observed in the liver and blood, along with mod-
ications in glutathione reductase and peroxidase.
94
Using
FRAP and DPPH radical scavenging techniques, the antioxidant
Figure 6. Chemical structure of quinones specic for C. ofcinalis.
37
.
Sapkota and Kunwar 11
Figure 7. Chemical structure of essential oil component specic for C. ofcinalis.
57
.
12 Natural Product Communications
capacityof the ethanolic extract of C. ofcinalis was evaluated. The
presence of avonoids, saponins, and polyphenolsall recog-
nized for their antioxidant qualitieswas veried by a prelimi-
nary phytochemical investigation. It was found that the IC50
for radical scavenging activity was 116 µg/mL.
95
Through its
ability to scavenge NO radicals, DPPH, and ABTS, ower
extract of C. ofcinalis has demonstrated notable antioxidant activ-
ity. Using the FRAP method, the mouthwash made from C. of-
cinalis was also assessed for its antioxidant activity and showed a
decreased IC50 value that was comparable to ascorbic acid.
94,96
Moreover, the FRAP method was used to assess the antioxidant
activity of C. ofcinalis oral extract with varying proportions of
water and ethyl alcohol. The hydro alcoholic extract (50:50 v/
v) exhibited the highest level of antioxidant activity.
In a different investigation, the antioxidant activity of silver
nano-particles biosynthesized from C. ofcinalis aqueous
extract was assessed using the DPPH, FRAP, and ABTS
methods. Using the ABTS technique, the studys strongest
Figure 8. Chemical structure of carotenoids specic for C. ofcinalis.
57,69
.
Sapkota and Kunwar 13
antioxidant activity was discovered. The values of antioxidant
activity ranged from 14.3 to 43.6 mg CGA g1DW.
97
In a
related investigation, high-performance liquid chromatography
was used to assess and quantify the antioxidant activity of C. of-
cinalis hydroalcoholic leaf extracts. The DPPH technique for
superoxide and hydroxyl radicals was used to evaluate antioxi-
dant capability. The plants signicant antioxidant activity was
demonstrated by the DPPH and hydroxyl methods in particu-
lar. This activity may have been caused by the plants identica-
tion of avonoids (24.67 mg/g), polyphenols (33.90 mg/g),
condensed tannins (27.30 mg/g), rutin (37.25 mg/g), and
quercetin (6.09 mg/g) in the study.
98
Using DPPH free
radical, hydroxyl radical, and lipid peroxyl radical as indicators
of effects, Cetkovic et al investigated the effects of methanolic
and water extracts from growing wild marigold (GWM),
Calendula arvensis L., and cultivated marigold at concentrations
ranging from 0.10 to 0.90 mg/ml. Cultivated marigold extracts
demonstrated stronger antioxidant and scavenging activity than
GWM extracts; water extracts performed better than metha-
nolic extracts. The most potent antioxidants were found in
the water extracts from cultivated marigold; at a concentration
of 0.75 mg/ml, they totally removed the hydroxyl radicals
Figure 9. Chemical structure of lipid and fatty acids specic for C. ofcinalis.
57
.
14 Natural Product Communications
produced in the Fenton system. This extract prevented 92% of
DPPH radicals and 95% of peroxyl radicals from forming
during lipid peroxidation at the same dose.
24
The substantial
concentration of polyphenols, avonoids, and tannins in the
plant extract may be the cause of its high antioxidant potential.
99
Anti-inammatory and Analgesic Activity
For many years, C. ofcinalis has been a mainstay in conventional
medicine, used to treat both human and animal pain and inam-
mation. This adaptable treatment is made in many formstinc-
tures, ointments, lotions, decoctions, and uid extractsfor a
range of uses. The plantsowers and leaves can be used to
make galenic medicines and pharmaceutical formulations that
have a variety of pharmacological effects, most notably analge-
sic and anti-inammatory properties.
100
An animal study veried calendula ower extracts ability to
effectively reduce inammation. NO, a pro-inammatory sub-
stance produced by NO synthase and strongly secreted by
innate immune cells during inammation, is inuenced by
ower extract of C. ofcinalis. Calendula ower extract was dis-
covered to display a dose-dependent suppression of NO, with a
50% decrease at 147 μL/mL without resulting in any harm to
cells. This offers signicant proof of the anti-inammatory
characteristics of calendula ower extract.
101
An assessment
was made of the analgesic and anti-inammatory qualities of
a hydroalcoholic extract derived from the aerial section of Pot
marigold. Acetic acid writhing tests and hot water tail immer-
sion were utilized to assess its analgesic effects. Additionally,
paw edema generated by carrageenan was used to test the
ability to prevent inammation. For the trials, adult male and
female Wistar rats weighing 200220 g and albino mice weigh-
ing 2030 g were employed. Additionally, the extracts showed a
dose-dependent effect on the suppression of inammation and
discomfort.
102
Using albumin denaturation assays, the anti-
inammatory properties of the C. ofcinalis tea formulation
were assessed. At 10 µl and 20 µl, the extract showed a substan-
tial anti-inammatory effect (P-values of 0.002 and 0.000,
respectively). It was discovered that the anti-inammatory activ-
ity outperformed the control group at every concentration
tested.
103
In another investigation, the anti-inammatory efcacy
of C. ofcinalis extracts was assessed using the carrageenan-
induced acute paw edema and dextran-induced acute paw
edema methods. In both approaches, the oral administration
of oral extract (250 and 500 mg/kg) resulted in a considerable
suppression of rat paw edema. Its anti-inammatory effect is
supported by the observation that pro-inammatory cytokines
such as IL-1beta, IL-6, TNF-alpha, and IFN-gamma, as well as
acute phase protein, C-reactive protein, were decreased in
mice.
104
C. ofcinalis oral oils were tested for their ability to reduce
inammation in response to LPS-stimulated macrophages.
The outcome demonstrates that calendula oil (147 µL/mL) dra-
matically (50%) decreased nitrite formation, and the presence of
other terpenoids, such as faradiol, is crucial for demonstrating
anti-inammatory effect.
101,105
The plants extract from the
leaves has analgesic properties as well. The analgesic efcacy
of pure and diluted C. ofcinalis leaf extract (1:1, 1:2) was
studied in mice following intraperitoneal injection. The pure
extract exhibited the highest degree of analgesic effectiveness
and the longest duration of action after thirty minutes of
injection.
106
Antibacterial Activity and Antifungal Activity
Numerous investigations were carried out to assess the antifun-
gal and antibacterial properties of C. ofcinalis. Using the agar
well diffusion method, it was discovered that C. ofcinalis
oral oil demonstrated strong antibacterial action against
Bacillus subtilis, Escherichia coli,Pseudomonas aeruginosa, and
Staphylococcus aureus.
107
The antibacterial activity of CO tincture
60% (v/v) was found signicant against Pseudomonas aeruginosa
Figure 10. Pharmacological action of C.ofcinalis.
Sapkota and Kunwar 15
by disc diffusion method.
108
The leaf powder of C. ofcinalis
showed signicant antibacterial activity in agar well diffusion
method against Bacillus subtulis, Staphylococcus aureus, Escherichia
coli, Candida glabrata, and Klebsiella pneumonia.
109-112
Similarly
Ghaima et al conducted antibacterial assay of water extract of
C. ofcinalis ower against Salmonella, Shigella dysenteriae, Shigella
exneri, Shigella sonnei,and E. coli by agar diffusion method in
which the adherent growth of bacterias were decreased.
112
A
study was conducted which compare the antimicrobial property
of the mouth wash that contain C. ofcinalis L., Camellia sinensis
(L.) Kuntze extract, and 0.12% chlorhexidine digluconate. It
was found that all mouth wash exhibited antimicrobial property
but they were not effective as 0.12% chlorhexidine digluconate.
The antibacterial activity of C. ofcinalis was supposed to be
exhibited by the presence of oleanolic acid,
113
alkaloids, glyco-
sides, avonoids, and terpenoids.
112
The antifungal activity of C. ofcinalis was evaluated against
A.niger, R. japonicum, Candida krusei, Candida tropicallis, Candida
parapsilosis, Candida dubliniensis, Candida albicans, Candida glabrata,
and R.glutinis fungus. The growth inhibition ranged from 10 mm
to 20 mm against those tested fungi and was comparable to
amphotericin B and nystatin. The antifungal activity of C. ofci-
nalis may be due to the presence of phytochemicals like cardiac
glycoside, sterols, and avonoids.
23,114
Anticancer Activity
The crystal violet assay and 3-(4,5-Dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) assay are some of the
methods to evaluate anticancer activity of C. ofcinalis plants.
The methanolic leaves extract was evaluated by crystal violet
assay against Michigan Cancer Foundation-7 (MCF-7),
Ahmed, Mahfoodha, Mortadha, Jabria-2013 (AMJ13), M.D.
Anderson-metastasis breast cancer (MDA-MB), and CAL51
breast cancer lines. The growth of cancer cell was inhibited by
extract lines while signicant inhibition was found in
MDA-MB cells.
115
The anti-colorectal cancer activity of silver
nano-particles of C.ofcinalis was evaluated against MTT
assays which decreased the colorectal cell lines.
4
The cytotoxic
activity of some isolated compounds like lutein, lupeol, and
eugenols were evaluated by MTT assays against breast cancer
cell lines (MCF-7 and MDA-MB-231) and normal breast cell
line (MCF 10A). The messenger ribonucleic acid (mRNA)
expression level of p53, caspase-3, and bax genes were mea-
sured which were found to increase in both cancer cells, and
B-cell lymphoma 2 (Bcl-2) gene expression deduced in treated
breast cancer cells.
116
The presence of major avonoids
which are avone and luteolin-7-O-β-glucoside which is used
in cancer therapy are present in C. ofcinalis and may be useful
for exhibiting anticancer activity.
117
Cardio-protective Effect
C. ofcinalis extracts are found to be effective against ischemic
heart disease. By altering antioxidant and anti-inammatory
pathways, cardio-protective activity is done by turning the ische-
mia reperfusion-mediated death signal into a survival signal, as
shown by the activation of protein kinase B (Akt) and Bcl2
and the reduction of TNF-α.
118
In another study, plant extract
(0.3 mg/l) inhibited the heart rate contractility by 100% which
may be due to spasmogenic activity of plant extract.
34,119
Antiulcer Activity
Pyloric ligation-induced ulcer model, ethanol-induced ulcer
model, cold-resistant ulcer and indomethacin-induced ulcer
model were widely used models to assess antiulcer activity of
C. ofcinalis.C. ofcinalis ethanolic extract decreased the gastric
acid effects on mucosa as compared to ranitidine in experimen-
tal rats.
120
The hydroalcoholic extract was found effective in
reducing the gastric acid secretion and prevent the effects of
aspirin in gastric mucosa.
121
Yadav et al investigated the antiulcer
efcacy of C. ofcinalis entire plants (250 and 450 mg/kg/b.w) and
discovered that the maximum treatment efciency (450 mg/kg b.w.)
in cold-resistant stress-induced ulcers was 87.15%. This was shown
to be suppressing the formation of ulcers generated by physical
and chemical agents.
122
The pharmacological properties of some isolated com-
pounds are as shown in Table 3.
Toxicological Activity
Rats and mice were given the hydroalcohol extract (HAE)
made from C. ofcinalis to evaluate its acute toxicity through
oral administration. Its subacute effects on biochemical, mor-
phologic, and hematological markers in rats were also
Table 3. List of Isolated Compounds and its Pharmacological Action.
S.N Compounds Pharmacological actions
1. Stigmasterol anticancer, anti-osteoarthritis,
anti-inammatory, antidiabetic,
immunomodulatory, antiparasitic,
antifungal, antibacterial, antioxidant, and
neuroprotective properties
123,124
2. Ψ-taraxasteol Antitubercular activity
56
3. Lupeol Anti-Inammatory activity
56
4. Calenduloside
B
Antiulcerous
125
5. Calenduloside Cytotoxicity against, melanoma leukemia and
colon cancer
56
6. Calendoaside Covid-19
126
, antiparasitic
127
7. Isorhamnetin anti-tumor, anti-inammatory, anti-oxidation,
organ protection, prevention of obesity
128
anti-acetylcholinesterase activity
47
8. Narcissin Cytotoxic activity
56
9. α-thujene Anti-inammatory, Antifungal
129
10. Carvone antidiabetic, anti-inammatory, anticancer,
neurological, antimicrobial, antiparasitic,
antiarthritic, anticonvulsant, and
immunomodulatory effects
130
16 Natural Product Communications
assessed. The rats in the acute toxicity test did not die when
oral dosages of HAE up to 5.0 g/kg were given. In compari-
son to the control group, oral administration of HAE at doses
of 0.025, 0.25, 0.5, and 1.0 g/kg did not result in any apprecia-
ble hematological alterations. However, elevated levels of
alanine transaminase (ALT) and blood urea nitrogen were
noted in terms of biochemical markers. Examinations of the
brain, kidney, and heart morphologically showed no obvious
changes.
131
Male and female Wistar rats were used to test
theacuteandsub-chronicoraltoxicitiesofC. ofcinalis
extract. Sub-chronic doses of 50, 250, and 1000 mg/kg/day
were given in drinking water, whereas a single acute dose of
2000 mg/kg dissolved in distilled water was given orally.
Numerous toxicological endpoints were assessed, such as
body weight, food and water consumption, tissue weights,
and results of histological analyses. Blood chemistry
(glucose, cholesterol, urea, proteins, ALT, aspartate amino-
transferase (AST), alkaline phosphatase) and blood elements
(hematocrit, hemoglobin, erythrocyte and leukocyte count,
clotting time) were also examined. There were no deaths or
toxicity indicators in the acute investigation. Nonetheless,
several blood componentshemoglobin, erythrocytes, leuko-
cytes, and clotting timewere substantially impacted in both
genders in the sub-chronic trial conducted 90 days later.
There was also an effect on blood chemistry measures such
as alkaline phosphatase, AST, and ALT. Histopathological
analysis identied little irregularities in the liver tissue that
matched the biochemical changes that had been seen. These
results collectively imply that there are less acute and sub-
chronic toxicity associated with.
132,133
Using organization for economic co-operation and develop-
ment (OECD) criteria 402 and 411, another study was carried
out to examine the acute and sub-chronic skin toxicity of calen-
dula essential oil in order todetermine its safety. Animals exposed
sub-chronically to C. ofcinalis essential oil for 90 days were given
doses of 2.5, 5, and 10 mL/kg, while animals exposed acutely to
20 mL/kg of the oil. The CNS effects, organ weight, necropsy,
biochemical and hematological variables, and histopathology
were among the parameters. Every animal exhibited typical
behavior, and there were no anomalies found in the necropsy, his-
tology, blood biochemistry, or hematology. It was discoveredthat
the CO oils no observed adverse effect level and no observed
effect level were 2.5 and 10 mg/kg/day, respectively. With an
LD50 value of 20 mL/kg body weight, C. ofcinalis oil categorized
by the European Medicines Agency as a herbal medical product
showed no appreciable harmful effects.
134
Industrial and Pharmaceutical Applications
C. ofcinalis extract is a botanical ingredient extensively utilized
in cosmetics for its soothing and skin-conditioning attributes.
It is a key component in various products such as creams,
balms, and serums, offering gentle and natural skincare solu-
tions. Commonly known as calendula oil, this extract provides
numerous benets, nurturing the skin and enhancing overall
complexion.
135
The market for C. ofcinalis ower extract wit-
nessed signicant growth, reaching USD 26 billion in 2023.
Projections indicate further expansion, with an expected value
of USD 64.43 billion by 2030, fueled by a compound annual
growth rate) of 30% from 2024 to 2030.
136
C. ofcinalis prepa-
rations commonly include carophyllenic ointment, which con-
tains carotenoids extracted from the owers, as well as pot
marigold tincture. Additionally, it serves as a component in pro-
prietary homeopathic medicines, utilized for addressing symp-
toms linked with acute musculoskeletal injuries such as pain
and swelling.
137
Furthermore, otic solution and naturopathic
herbal extract ear drops, formulations derived from naturo-
pathic principles containing Calendula owers, have demon-
strated efcacy in managing otalgia associated with acute
otitis media in children.
8,138
Furthermore, the exploration of novel drug delivery systems
incorporating C. ofcinalis extract is still in its early stages, with
on-going research anticipated in this domain. Molecular docking
and molecular dynamics represent modern computational tech-
niques in drug design, holding signicant promise for the develop-
ment of new therapeutic candidates across various health
conditions. By analyzing the chemical composition of medications
and their target receptors, the therapeutic potential of numerous
bioactive compounds can be investigated, offering potential
savings in time and resources. In the foreseeable future, formula-
tions containing C. ofcinalis at micro- and nano-levels demonstrate
promising prospects for treating various ailments, with anticipated
advancements and applications expected to be remarkable.
Additionally, combining C. ofcinalis with other available agents
for activity enhancement presents a promising strategy that
could ultimately improve pharmacological outcomes.
139
Conclusions
C. ofcinalis is one of the most potent and benecial owering
plant with multi health benets and pharmacological actions. It
contains different chemical constituents like: terpenoids, glyco-
sides, avonoids, volatile oil, and carbohydrates that results dif-
ferent physiological actions in the body. Various clinical studies
have also proven the useful therapeutic activities like: antioxidant,
antidiabetic, hypolipidemic, anti-inammatory, hepatoprotective,
and anticancer. These all activities make the plant rich in bioactiv-
ities and even depict the remarkable possibilities of the plant in
the eld of research and medicine. Numerous studies have
been performed and showed the owers extract contained the
greater biological activities in comparison to the other parts.
Furthermore, comprehensive study in relation to the safety and
efcacy as well as toxicity should be performed for the proper
designing of the formulations of the plant in the future.
Declaration of Conicting Interests
The authors declared no potential conicts of interest with respect to
the research, authorship, and/or publication of this article.
Sapkota and Kunwar 17
Funding
The authors received no nancial support for the research, authorship,
and/or publication of this article.
ORCID iDs
Biswash Sapkota https://orcid.org/0000-0002-5272-9122
Paridhi Kunwar https://orcid.org/0009-0006-1184-1923
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22 Natural Product Communications
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Objective The present study evaluated the antinociceptive activity of Calendula officinalis L. (Ca) cream on inflammatory hypernociception. Methods Creams with different Ca concentrations were tested for their ability to ameliorate pain-related behavior and edema in rats using formalin test, carrageenan (Cg)-induced acute inflammation model, bradykinin (BK)-induced acute inflammation model, and complete Freund’s adjuvant (CFA)-induced chronic inflammation model. Using the formalin test, we also examined the implication of peripheral opioid receptors in the antinociceptive mechanisms of Ca cream, by means of Q-naloxone, a peripherally acting nonselective opioid antagonist. Furthermore, the effects of Ca cream compared with diclofenac on BK-induced edema were examined when the kininase II in tissue was preserved or inhibited by captopril. The local production of redox biomarkers in formalin model, tumor necrosis factor-α (TNF-α) in Cg model and histopathological changes in CFA model were also evaluated. Results A single application of Ca cream at a dose of 10% or 30% (w/w) decreased the formalin-induced licking/biting behavior during both phases of this test in a Q-naloxone-sensitive manner. This effect was associated with the reduction of oxidative stress in the injured paw and the edema associated with the later phase of formalin-induced pain. A single application of Ca cream (10% or 30%) reduced paw edema and thermal hypernociception in Cg-induced acute inflammation, corresponding with a local decrease in TNF-α. Ca cream also inhibited BK-induced spontaneous nociceptive behavior and paw inflammation in both paradigms studied. Repeated applications of Ca cream also decreased CFA-induced chronic inflammation, mechanical hypernociception and histopathological changes in the paw. Conclusion These results reveal the topical antinociceptive and antiedematogenic effects of Ca cream. A modulatory action on peripheral opioid receptors associated with its antioxidant mechanism may be involved, at least in part, in its analgesic effects. These findings may have an impact on the clinical management of painful inflammatory diseases.
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