Content uploaded by Sulaiman C T
Author content
All content in this area was uploaded by Sulaiman C T on Jan 12, 2021
Content may be subject to copyright.
R E S E A R C H Open Access
Chemical profiling of selected Ayurveda
formulations recommended for COVID-19
Sulaiman C. T.
1*
, Deepak M.
1
, Ramesh P. R.
2
, Mahesh K.
2
, Anandan E. M.
3
and Indira Balachandran
1
Abstract
Background: The novel coronavirus disease 2019 (COVID-19), caused by the severe acute respiratory syndrome
coronavirus-2 (SARS-CoV-2), is the global health concern since December 2019. It has become a big challenge for
the researchers to find a solution for this newly evolved pandemic. In Ayurveda point of view, COVID-19 is a
Janapadodhwamsa vikara (epidemic disease), a situation where the environment—air, water, land, and seasons—is
vitiated, causing a simultaneous manifestation of a disease among large populations. The aim of this study is to
identify the active compounds of selected Ayurveda medicines recommended for COVID-19.
Results: The selected preparations are traditionally recommended for the management of various kinds of fever
including the infectious ones and to enhance the immunity. HPTLC analysis of the same showed presence of many
active molecules like umbelliferone, scopoletin, caffeic acid, ferulic acid, gallic acid, piperine, curcumin, berberine,
and palmatine.
Conclusion: The study provided valuable scientific data regarding the active ingredients of the selected medicines
with proven therapeutic potentials like anti-viral, immunomodulatory, and anti-inflammatory activities.
Keywords: COVID-19, Ayurveda, Indukantham Kwatham, Vilvadi Gulika, Mukkamukkatuvadi Gulika, HPTLC
1 Background
COVID-19 has emerged as the most dangerous global
pandemic threat since its outbreak during December
2019 in Wuhan, China. As of September 4, 2020, the
World Health Organization (WHO) has reported more
than 26 million confirmed cases and 8.6 lakhs deaths
worldwide and it has spread to 216 countries, areas, or
territories (https://www.who.int/emergencies/diseases/
novel-coronavirus-2019). Now, it is a big challenge for
the researchers and health professionals to find out a so-
lution for this deadly viral infection. COVID-19 is a viral
infection that has been known to have the fastest fre-
quency of replication in its positive strand resulting in
the quick development of new progeny viral cells inside
the host cells. SARS-CoV-2 is a single-stranded RNA
pathogen, which is characterized by a high mutation rate
[1,2]. In Ayurveda point of view, COVID-19 is a Jana-
padodhwamsa vikara (epidemic disease), a situation
where the environment—air, water, land, and seasons—
is vitiated, causing a simultaneous manifestation of a dis-
ease among large populations [3].
Medicinal plants have been used as a treatment and
defensive strategy for several infectious diseases since
ancient times. The benefit of using these herbs in viral
respiratory infections is to build immune-stimulating
and inflammation-modulating effects to prevent severe
life-threatening conditions. Holistic approach of Ayur-
veda focuses on prevention of diseases through lifestyle
modification, dietary management, prophylactic inter-
ventions for improving the immunity, and managing the
symptoms using herbal preparations. Medicinal plants
have been reported to have anti-viral activity and many
species such as Aegle marmelos,Andrographis panicu-
lata,Acacia nilotica,Ocimum tenuiflorum,Piper nigrum,
Solanum nigrum, and Terminalia chebula have been sci-
entifically proved for their anti-viral properties [4–6].
© The Author(s). 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License,
which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give
appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if
changes were made. The images or other third party material in this article are included in the article's Creative Commons
licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons
licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain
permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
* Correspondence: slmnct@gmail.com;sulaimanct@aryavaidyasala.com
1
Phytochemistry Division, Centre for Medicinal Plants Research, Arya Vaidya
Sala, Kottakkal, Malappuram, Kerala 676503, India
Full list of author information is available at the end of the article
Beni-Suef University Journal o
f
Basic and Applied Sciences
C. T. et al. Beni-Suef University Journal of Basic and Applied Sciences
(2021) 10:2
https://doi.org/10.1186/s43088-020-00089-1
Ayurveda medicines were recommended by the Ministry
of AYUSH, Government of India to enhance the immunity
and to prevent the severe conditions of Cov-2 infection.
Detailed guidelines have been published by the AYUSH
Ministry regarding the management of COVID-19 (https://
www.ayush.gov.in/ayush-guidelines.html). About 80% of
COVID-19 cases are with mild symptoms requiring only
primary medical care. Ayurveda medicines are advised to
patients with mild symptoms and those under surveillance
which addresses the therapeuticprovincewithinanintegra-
tive model of care [7]. The present study was focused on
the identification of active ingredients of certain Ayurveda
medicines such as Indukantham Kwatham (IK), Vilvadi
Gulika (VG), and Mukkamukkatuvadi Gulika (MMG) in
which the ingredient plants have been reported to possess
immunomodulatory and anti-viral properties.
Indukantham Kwatham is a polyherbal tablet prepared
out of specific parts of different medicinal plants such as
Holoptelea integrifolia,Cedrus deodara,Gmelina
arborea,Aegle marmelos,Stereospermum colais,Oroxy-
lum indicum,Premna corymbosa,Desmodium gangeti-
cum,Pseudarthria viscida,Solanum anguivi,Solanum
virginianum,Tribulus terrestris,Piper longum,Piper
mullesua,Plumbago zeylanica, and Zingiber officinale.It
Fig. 1 HPTLC profiling of IK, MMG, and VG documented at UV-254, 366, and visible light
C. T. et al. USERDEF_JournalTitle> (2021) 10:2 Page 2 of 5
is generally used for the treatment of intermittent fever
and fatigue and to enhance the resistance power [8].
Vilvadi Gulika is prepared using different parts of the
various medicinal plants such as Aegle marmelos,Ocimum
tenuiiflorum,Pongamia pinnata,Veleriana jatamansi,
Cedrus deodara,Terminalia chebula,Phyllanthus emblica,
Terminalia bellirica, Zingiber officinale,Piper nigrum,Piper
longum,Curcuma longa,andBerberis aristata. The ingredi-
ent plants of Mukkamukkatuvadi Gulika are Terminalia
chebula,Phyllanthus emblica,Terminalia bellirica,Zingiber
officinale,Piper nigrum,Piper longum,Cuminum cyminum,
Nigella sativa,Acorus calamus,Swertia chirata,Cinnamo-
mum camphora,Myristica fragrans,Aloe vera,Syzygium
aromaticum,Allium sativum,Piper cubeba, Saussurea cos-
tus,Cinnamomum verum,Ferula assa-foetida,Trchysper-
mum roxburghianum,andVitex negundo [8].
2 Methods
2.1 Chemicals and reagents
Chemicals such as toluene (CN: IE5F650118), ethyl acet-
ate (CN: ID5F650128), and methanol (CN: SA5P650021)
were procured from Merck India.
2.2 Sample preparation
The selected medicines were obtained from the Product
Development Department of the Institute. Two grams
each of IK (Batch No. 198339), VG (Batch No. 193083),
and MMG (Batch No. 194967) were sonicated with
chromatographic grade methanol in an ultra-sound bath
(Labnet Scientific, India) for 20 min. It was then filtered
through a membrane filter (0.45 μm) and kept under the
refrigerator until HPTLC analysis.
2.3 Instruments and general chromatographic conditions
HPTLC analysis was carried out by the CAMAG HPTL
C system (Switzerland). Samples were applied using
CAMAG ATS 4 auto sampler on aluminum backed pre-
coated silica gel 60F
254
HPTLC plate (Merck India).
Mobile phase was optimized as toluene, ethyl acetate,
and methanol in the ratio of 7:3:1. The chromatogram
was developed in a saturated Twin Trough chromato-
graphic chamber (Camag, Switzerland) and was visual-
ized under UV-chamber (254 and 366 nm) and in visible
light after derivatizing with anisaldehyde sulfuric acid re-
agent followed by heating at 105 °C for 5 min.
3 Results
Rapid chromatographic method has been developed for
the chemical fingerprinting of selected medicines by
modern high-performance thin-layer chromatography.
The optimized mobile phase provided good resolution
under various documentation systems such as UV-254,
366, and visible light. Chromatogram and 3D-illustrated
display are presented in Fig. 1. HPTLC analysis showed
presence of various compounds belonging to different
groups of phytochemicals such as alkaloids, coumarins,
and phenolics. Structural identification was confirmed
with matching R
f
of standard compounds. The com-
pounds identified from the tested formulations are given
in Table 1.
Coumarins such as umbelliferone and scopoletin are
detected in all the three selected medicines and these
are the plant coumarins reported from many medicinal
plants. Phenolic compounds like caffeic acid and ferulic
acid are found in both IK and VG. Gallic acid was iden-
tified from all the three selected medicines and has been
reported from many ingredient plants. VG showed the
presence of alkaloids such as curcumin, berberine, and
palmatine which might have extracted from its ingredi-
ent plants like Piper nigrum,Piper longum,Curcuma
longa, and Berberis aristata.
4 Discussion
Bioactive compounds from natural products are attractive
candidates for drug development. Numerous medicinal
plants have been reported to possess various therapeutic
properties including anti-viral, anti-inflammatory, and im-
munomodulatory activities. The chemical profiling of
three selected medicines showed the presence of various
biologically active compounds belonging to different clas-
ses of phytochemicals such as alkaloids, phenolics, and
coumarins. Coumarins such as umbelliferone and scopole-
tin were found to be common for all the selected samples.
Naturally occurring coumarins have been reported to pos-
sess diverse biological and pharmacological properties such
as anti-viral, anti-coagulant, anti-bacterial, anti-fungal, anti-
protozoal, insecticidal, fungicide, anti-mycobacterial, anti-
mutagenic, anti-amnesic, and anti-inflammatory activities
[9,10]. There are numerous evidences for the inhibitory
role of coumarins against infection of various viruses such
as HIV, influenza, enterovirus 71, and coxsackievirus A16.
The mechanisms involve either inhibition of proteins essen-
tial for viral entry, replication, and infection or regulation of
Table 1 Compounds identified from IK, MMG, and VG by HPTLC
analysis
Sl. no. Compounds R
f
value Present in
1 Caffeic acid 0.21 IK, VG
2 Palmatine 0.33 VG
3 Ferulic acid 0.37 IK, VG
4 Gallic acid 0.43 IK, VG, MMG
5 Scopoletin 0.44 IK, VG, MMG
6 Berberine 0.45 VG
7 Umbelliferone 0.57 IK, VG, MMG
8 Piperine 0.71 MMG
9 Curcumin 0.73 VG
C. T. et al. USERDEF_JournalTitle> (2021) 10:2 Page 3 of 5
cellular pathways such as Akt-Mtor, NF-κB, and anti-
oxidative pathways including NrF-2 [11].
Alkaloids such as piperine, curcumin, berberine, and
palmatine were identified from the selected medicines.
Piperine is detected from both IK and MMG and that
might have come from the ingredient plant Piper
nigrum. The immunomodulatory potential of piperine
has been reported earlier [12]. Piperine was reported to
inhibit proliferative response induced by lipopolysac-
charide (LPS) and immunoglobulin α-IgM antibody and
resulted in inhibition of IgM antibody secretion and re-
duced expression of cluster of differentiation CD86 [13].
Another study by Lee et al. 2018 [14] demonstrated that
piperine in combination with gamma-aminobutyric acid
(GABA) mediated p38 and JNK MAPK activation, which
increased EPO and EPO-R expression, resulting in up-
regulation of IL-10 and NF-κB. Alkaloids like curcumin,
berberine, and palmatine were identified from VG. The
anti-viral effect of curcumin on Zika and chikungunya
viruses has been well established [15]. The literature
showed that curcumin mediates its anti-viral activity
through various mechanisms. Curcumin has been reported
to inhibit the Japanese encephalitis virus by dysregulated
ubiquitin-proteasome system and an accumulation of ubi-
quitinated proteins [16].Curcuminwasalsoreportedtoin-
hibit various virus replications like Rift Valley fever virus
and hepatitis C virus [17,18]. Moreover, curcumin was
shown to impact HCV replication through binding and fu-
sion [19] and similar results were reported in the case of
ZIKV and CHIKV. HIV-1 integrase activity of curcumin
was also reported previously [20]. Various pharmacological
activities of berberine such as anti-oxidant, anti-bacterial,
anti-inflammatory, anti-viral, nephroprotective, and cardio-
protective have been reported earlier [21]. Anti-viral activity
of berberine against human cytomegalovirus has been
reported previously [22]. The immunomodulatory ef-
fect of berberine was validated in many previous liter-
atures [23–25].
Currently, there are no available vaccines or specific
medicines for the treatment of COVID-19. In light of
the outbreak, various treatment modalities have been
considered, including herbal medicine, which has been
widely used during the past epidemic outbreaks, such as
severe acute respiratory syndrome (SARS) and H1N1 in-
fluenza. The phytochemicals identified from IK, MMG,
and VG are active molecules with potential biological
properties such as anti-viral, anti-inflammatory, and im-
munomodulatory activities.
5 Conclusion
COVID-19 pandemic is a global challenge for human
health, and researchers are urgently seeking medicine
for it. Currently, the treatment options for COVID-19
are limited due to non-availability of vaccines or specific
medicines. In this context, the search for traditional
herbal medicine is also a viable strategy for COVID-19
management. The present study on selected Ayurveda
medicines provided valuable scientific data regarding the
active ingredients of the drugs tested with proven thera-
peutic potentials like anti-viral, immunomodulatory, and
anti-inflammatory activities.
Abbreviations
IK: Indukantham Kwatham; VG: Vilvadi Gulika; MMG: Mukkamukkatuvadi
Gulika; HPTLC: High-performance thin-layer chromatography
Acknowledgements
SCT sincerely thank Dr. Geetha S Pillai, Deputy Project Director, CMPR, Arya
Vaidya Sala, Kottakkal, Kerala, India, for the valuable suggestions.
Authors’contributions
All authors have read and approved the manuscript. SCT: Designed and
implemented the work, done the data analysis; DM: Carried out HPTLC
analysis; RPR: Provided resources; MK: Contributed background data; AEM:
Prepared the samples; IB: Supervision and edited the manuscript.
Funding
This work was funded by Navajbhai Ratan Tata Trust, Mumbai, India (Grand
Number: RLC-PPP-AVS-20150930). The fund was used for required chemicals
and reagents.
Availability of data and materials
The datasets used and/or analyzed during the current study are available
from the corresponding author on reasonable request.
Ethics approval and consent to participate
Not applicable
Consent for publication
Not applicable
Competing interests
The authors declare that they have no competing interests.
Author details
1
Phytochemistry Division, Centre for Medicinal Plants Research, Arya Vaidya
Sala, Kottakkal, Malappuram, Kerala 676503, India.
2
Clinical Research
Department, Arya Vaidya Sala, Kottakkal, Kerala, India.
3
Product Development
Department, Arya Vaidya Sala, Kottakkal, Kerala, India.
Received: 12 August 2020 Accepted: 26 November 2020
References
1. Chitranshi N, Gupta VK, Rajput R, Godinez A, Pushpitha K, Shen T, Mirzaei M,
You Y, Basavarajappa D, Gupta V, Graham SL (2020) Evolving geographic
diversity in SARS-CoV2 and in silico analysis of replicating enzyme 3CLpro
targeting repurposed drug candidates. J Transl Med 18:278
2. Chen H, Guo J, Wang C, Luo F, Yu X, Zhang W, Li J, Zhao D, Xu D, Gong Q
(2020) Clinical characteristics and intrauterine vertical transmission potential
of COVID-19 infection in nine pregnant women: a retrospective review of
medical records. Lancet 395:809–815
3. Girija LT, Sivan N (2020) Ayurveda treatment of COVID-19/ SARS-CoV-2: a
case report. J Ayu Int Med. https://doi.org/10.1016/j.jaim.2020.06.001
4. Venkateswaran PS, Millman I, Blumberg BS (1987) Effects of an extract from
Phyllanthus niruri on hepatitis B and woodchuck hepatitis viruses: in vitro
and in vivo studies. Proc Natl Acad Sci U S A 84:274–278
5. Hudson JB (1990) Antiviral compounds from plants. CRC Press, Boca Raton,
Ann Arbor, Boston
6. Vellingiri B, Jayaramayya K, Iyer M, Narayanasamy A, Govindasamy V,
Giridharan B, Ganesan S, Venugopal A, Venkatesan D, Ganesan H,
Rajagopalan K, Rahman PKSM, Cho SG, Kumar NS, Subramaniam MD (2020)
C. T. et al. USERDEF_JournalTitle> (2021) 10:2 Page 4 of 5
COVID-19: a promising cure for the global panic. Sci Total Environ. https://
doi.org/10.1016/j.scitotenv.2020.138277
7. Rastogi S, Pandey DN, Singh RH (2020) COVID-19 pandemic: a pragmatic
plan for Ayurveda intervention. J Ayurveda Integr Med. https://doi.org/10.
1016/j.jaim.2020.04.002
8. Kurup KV (2017) Sahasrayogam [Text book of 1000 Herbal formulations]
Malayalam translation with Vaidya Priya Commentary. Sree Rama Vilasam
Press and Book Depo, Kollam, pp 446–457
9. Karatas MO, Uslu H, Alici B, Gokce B, Gencer N, Arslan O (2016) Some
coumarins and benzoxazinones as potent paraoxonase 1 inhibitors. J
Enzyme Inhib Med Chem:1386–1391
10. Sulaiman CT, Deepak M, Sunil AR, Lijini KR, Balachandran I (2019)
Characterization of coumarins from Ipomoea mauritiana Jacq by LC-APCI-
MS/MS analysis and evaluation of its anti-amnesic activity. Beni-Suef Univ J
Basic Appl Sci https://doi.org/10.1186/s43088-019-0022-z
11. Mishra S, Pandey A, Manvati S (2020) Coumarin: an emerging antiviral
agent. Heliyon. https://doi.org/10.1016/j.heliyon.2020.e03217
12. Bezerra DP, De Castro FO, Alves APNN, Pessoa C, De Moraes MO, Silveira ER,
Lima MAS, Elmiro FJM, De Alencar NMN, Mesquita RO (2008) In vitro and
in vivo antitumor effect of 5-FU combined with piplartine and piperine. J
Appl Toxicol 28:156–163
13. Bernardo AR, Da Rocha JDB, De Lima MEF, Ricardo DD, Da Silva LHP,
Peçanha LMT, Danelli MDGM (2015) Modulating effect of the piperine, the
main alkaloid from Piper nigrum Linn., on murine B lymphocyte function.
Braz J Vet Med 37:209–216
14. Lee YM, Choi JH, Min WK, Han JK, Oh JW (2018) Induction of functional
erythropoietin and erythropoietin receptor gene expression by gamma-
aminobutyric acid and piperine in kidney epithelial cells. Life Sci 215:207–
215
15. Mounce BC, Cesaro T, Carraua L, Vallet T, Vignuzzi M (2017) Curcumin
inhibits Zika and chikungunya virus infection by inhibiting cell binding.
Antivir Res. https://doi.org/10.1016/j.antiviral.2017.03.014
16. Dutta K, Ghosh D, Basu A (2009) Curcumin protects neuronal cells from
Japanese encephalitis virus-mediated cell death and also inhibits infective
viral particle formation by dysregulation of ubiquitin-proteasome system. J
NeuroImmune Pharmacol:328–337
17. Kim K, Kim KH, Kim HY, Cho HK, Cheong SJ (2010) Curcumin inhibits
hepatitis C virus replication via suppressing the Akt-SREBP-1 pathway. FEBS
Lett 584:707–712
18. Narayanan A, Kehn-Hall K, Senina S, Lundberg L, Duyne RV, Guendel I, Das
R, Baer A, Bethel L, Turell M, Hartman AL, Das B, Bailey C, Kashanchi F (2012)
Curcumin inhibits Rift Valley fever virus replication in human cells. J Biol
Chem 287:33198–33214
19. Anggakusuma CCC, Schang LM, Rachmawati H, Frentzen A, Pfaender S,
Behrendt P, Brown RJP, Bankwitz D, Steinmann J, Ott M, Meuleman P, Rice
CM, Ploss A, Pietschmann T, Steinmann E (2014) Turmeric curcumin inhibits
entry of all hepatitis C virus genotypes into human liver cells. Gut:631137–
631149
20. Mazumder A, Raghavan K, Weinstein J, Kohn KW, Pommier Y (1995)
Inhibition of human immunodeficiency virus type-1 integrase by curcumin.
Biochem Pharmacol 49:1165–1170
21. Neag MA, Mocan A, Echeverría J, Pop RM, Bocsan CI, Crisan G, Buzoianu AD
(2018) Berberine: botanical occurrence, traditional uses, extraction methods,
and relevance in cardiovascular, metabolic, hepatic, and renal disorders.
Front Pharmacol. https://doi.org/10.3389/fphar.2018.00557
22. Hayashi K, Minoda K, Nagaoka Y, Hayashi T, Uesato S (2007) Antiviral activity
of berberine and related compounds against human cytomegalovirus.
Bioorg Med Chem Lett 17:1562–1564
23. Li H, Li XL, Zhang M, Xu H, Wang CC, Wang S (2014) Berberine ameliorates
experimental autoimmune neuritis by suppressing both cellular and
humoral immunity. Scand J Immunol 79:12–19
24. Ransohoff RM, Hafler DA, Lucchinetti CF (2015) Multiple sclerosis —a quiet
revolution. Nat Rev Neurol:11134–11142
25. Liu X, Zhang X, Ye L, Yuan H (2016) Protective mechanisms of berberine
against experimental autoimmune myocarditis in a rat model. Biomed
Pharmacother 79:222–230
Publisher’sNote
Springer Nature remains neutral with regard to jurisdictional claims in
published maps and institutional affiliations.
C. T. et al. USERDEF_JournalTitle> (2021) 10:2 Page 5 of 5
