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The Pharma Innovation Journal 2018; 7(6): 286-289
ISSN (E): 2277- 7695
ISSN (P): 2349-8242
NAAS Rating: 5.03
TPI 2018; 7(6): 286-289
© 2018 TPI
www.thepharmajournal.com
Received: 12-04-2018
Accepted: 13-05-2018
Sadhana Singh
Senior Resident & Phd Scholar in
Dept. of Dravyaguna, I.M.S,
B.H.U, Varanasi, Uttar Pradesh,
India
Apurva Priyadarshi
Phd Scholar in Dept. of
Dravyaguna, I.M.S, B.H.U,
Varanasi, Uttar Pradesh, India
Brijesh Singh
Assist. Professor in Dept. of
Shalya Tantra, Govt. Ayurveda
College Bilaspur, Chhattisgarh,
India
Poonam Sharma
Assist. Professor in Dept. of
Dravyaguna Apex institute of
Ayurveda Medical Science,
Varanasi, Uttar Pradesh, India
Correspondence
Brijesh Singh
Assist. Professor in Dept. of
Shalya Tantra, Govt. Ayurveda
College Bilaspur, Chhattisgarh,
India
Pharmacognostical and phytochemical analysis of
Pippali (Piper longum Linn.)
Sadhana Singh, Apurva Priyadarshi, Brijesh Singh and Poonam Sharma
Abstract
Ayurveda is the science and philosophy of life. Ayurveda reveals that we had a rich knowledge of the
application of medicinal plants from very ancient times. Plants have played a significant role in
maintaining human health and improving the quality of human life for thousands of years and have
served human as well as valuable components of medicines, seasoning, beverages, cosmetics and drugs.
In this study Botanical description, chemical Constituents, Ayurvedic Properties of Pippali, medicinal
properties, formulation, dose, and pharmacological activity of Piper longum Linn. are described. The
present work deals with the pharmacognostical and preliminary phytochemical studies on Fruit of Piper
longum Linn. Macroscopical and Microscopical Characters, physico-chemical constants, quantitative
microscopy parameters, extractive values, TLC and HPTLC were studied. Preliminary phytochemical
screening on Fruit of Piper longum Linn. were studied. The determination of these study will help future
researchers in their Phytochemical as well as Pharmacological evaluation of this plant.
Keywords: Piper longum Linn. preliminary phytochemical, pharmacognostical study
Introduction
Botanical Description
Habitat: It occurs in the hotter parts of India, from the central Himalayas to Assam, the khasi
& the mikir hills, the lower hills of Bengal [1] & the evergreen forests of the Western Ghats
from konkan to Travancore
Habit: A slender, aromatic climber with perennial woody roots.
Root: Root stock erect, thick, jointed, and branched.
Stem: Stems numerous, 0.6 -0.9m, ascending or prostrate (not climbing) much branched,
stout, cylindrical, thickened above nodes, finely pubescent.
Leaves: Ovate, cordate, subacute, entire, glabrous.
Flowers: Flowers are unisexual, minute, sessile bracteate without perianth very densely
packed in spikate inflorescences. The male & female spikes are on separate plants.
Fruit: Ovoid, yellowish orange, sunk in fleshy spike [2].
Pippali of Pippali Dry fruit
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Chemical Constituents
Two alkaloids piperlongumine & piperlonguminine, n-
hexadecane, n- heptadecane, n octadecane, n- nonadecane n-
eicosane, n-heneicosane, - thujene, terpinolene, zingiberene,
p-cymene, p-methoxy acetophenone, traces of dihydrocarveol,
phenyl ethyl alcohol & two sesquiterpenes; piperine, piplartin,
triacontane, dihydrostigmasterol, an unidentified setroid,
reducing sugar, glycosides, sesamin & methyl- 3.4.5-
trimethoxycinnamate (root) ; major alkaloid piperine &
sesamin (stem & fruit); sesquiterpene hydrocarbon,
caryophyllene, a sesquiterpene alcohol, carbonyl compound
(essential oil), N-isobutyldeca- trans- 2-trans- 4- dienamide,
piperine, piplartine & a lignan d- sesamin, two piperidine
alkaloids pipernonaline & piperundecalidine (fruit), sylvatin
sesamin & diaeudesmin (seed) [2].
Ayurvedic Properties of Pippali [3]
Rasa : Katu, Tikta, Madhura.
Guna : Laghu, Snigdha.
Virya : Anushna.
Vipaka : Madhura.
Doshakarma : Kapha vata shamaka
Rogaghnata
Kasa, Shvasa, Hikka, Aruci, Agnimandya, Ajirna, Vibandha,
Gulma, Udara vikara, Arsha, Yakridvikara, Pliha vriddhi,
Krimiroga, Hriddourbalya, Pandu, Raktavikara, Amvata,
Vatarakta, Kshaya, Yakshma, Shukradourbalya, Rajorodha,
Kashtaprasava, Kushtha, Jirna jvara, Vishamajvara,
Dourbalya5,6,7,8.
Karma
Kasahara, Shvasahara, Hikkanigrahana, Rasayana Medhya,
Vatahara Mutrala, Dipana, Vatanulomana, Shulaprashamana,
Mridurecana, Krimighna, Raktotkleshaka, Jantughna,
Shrovirecana, Plihavriddhihara, Yakriduttejaka,
Vishamajvara, Raktotkleshaka, Vrishya, Balya,
Garbhashyasamkocaka [8].
Prayojyanga : Phala, mula [10]
Dose : Curna 500mg-1gm [10]
Pharmacological activities
Antibacterial, anti-inflammatory, insecticidal, antimalarial,
CNS stimulant, ntitubercular, anthelmintic, hypoglycemic,
antispasmodic, cough suppressor, antigiardial,
immunostimulatory, hepatoprotective, analeptic, antinarcotic,
antiulcerogenic.
Formulations (Yoga)
Gudha Pippali, Pippalyasava, Pippali khandha, Pippalyadi
leha, Pippalyadi curna, Pippali rasayanam, Vardhamana
Pippali, Pippali ghritam.
Propagation and cultivation
Piper longum is cultivated on a large scale in limestone soil,
450-600m below the Cherrapunji region which receives very
heavy rains from the end of March to the middle of
September and where the relative humidity is high. Long
pepper is cultivated mainly by layering of mature branches or
by suckers planted at the beginning of the rainy season. The
vines are well manured with cowdung cake and start bearing
3-4 years after planting. The spikes are harvested in January,
while still green and unripe, as they are most pungent at this
stage. They are dried in the sun when they turn grey.
Plant regeneration from callus cultures of Piper longum Linn.
was achieved through organogenesis. In vitro grown shoots
were used as explants for callus injunction. Competent callus
was initiated around the nodal ring of tissue using Murashige
and Skoog's medium supplemented with 1.0 mg/1œ-
naphthaleneacetic acid and 0.2 mg/1, N-benzyladenine.
Optimum growth regulator concentration for shoot induction
and shoot elongation were found to be 0.5mg/1 indole -3-
acetic acid with 1.5mg/1benzyladenine and 0.1mg/1 indole-3-
acetic acid with 0.2 mg/1 benzyladenine respectively.
Elongated shoots were rooted on half strength Murashige and
Skoog's medium having 0.1mg/1 indole-3-acetic acid. The
rooted plantlets were successfully established in soil.
Morphogenetic potential of root, leaf, node and internode
explants of P. longum has been reported. The highest number
of shoot buds was produced on root explants followed by
node, internode and leaf explants. Benzyladenine was suitable
for shoot induction and its optimum concentration is 1-2 mg /l.
Material and Method
Microscopical Characteristics of Powdered Pippali (Friut)
Fragments of thin walled cell and starch grains were seen
from the powder of PippalÍ fruit. Coarse powder of dull dark
brown to black in colour. In microscopic powder study it
shows Lignified sclerenchymatous cells of pink to purple
colour and calcium oxalate crystals of grey in colour.
Aleurone grains of green colour and oil globules of orangish
pink stain
S. No.
Reagents
Observations
Characteristics
1.
Phloroglucinol+Conc. Hcl
Pink
Lignified sclerenchymatous cells
2.
Dil. Iodine solution
Blue
Starch present in perisperm
3.
Dil. Sulpuric Acid
White
Calcium oxalate crystals
4.
Alcoholic Picric Acid
Yellow
Starch grains
5.
Sudan Red III
Red
Oil globules and cuticles.
Table 1: Certificate of Analysis of Pippali
S. No.
Parameters
Observation
I
Physical tests
Nature
Coarse powder
Colour
Dull dark brown to black
Odour
Characteristic
Taste
Bitter
II
Foreign matter
Nil
III
Moisture content (w/w%)
9.34
IV
Ash value (% w/w)
Total ash
4.4
Acid insoluble ash
0.05
Water soluble ash
2.077
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Table 2: Percentage yield of Extracts of Pippali
S. No.
Extracts
Nature of
Extract
Weight
(gm)
% Yield w/w
I
Hydro-alcohol
Viscous
18.117
18.117
Preliminary phytochemical testing of Pippali (Piper
longum Linn.)
Genuine sample of Pippali gave the presence of following
Phytochemicals:
Phytoconstituents
Alkaloids
+
Glycosides
-
Flavonoids
-
Steroid
-
Phenolic & tannins
-
Terpenoid
-
Sterol
+
Carbohydrates
-
Proteins
+
Amino Acids
-
(+) indicate present, (-) indicate absent
Thin Layer Chromatography (TLC) of Extracts
TLC or Thin Layer Chromatography is a type of planar
chromatography. TLC is routinely used by researcher in the
field of phyto-chemicals, biochemistry etc. to identify the
components in a compound mixture like alkaloids,
phospholipids, amino acids etc. It is a semi quantitative
method of analysis and its sophisticated version or highly
precise quantitative version is High performance thin layer
chromatography (HPTLC). Similar to other chromatographic
methods TLC is also based on the principle of separation. The
separation depends on the relative affinity of compounds
towards stationary and mobile phase. The compounds that
under the influence of mobile phase (driven by capillary
action) travel over the surface of stationary phase. During this
movement the compounds with higher affinity to stationary
phase travel slowly while the others travel faster. Thus
separation of components in the mixture is achieved. Once
separation occurs individual components are visualized as
spots at respective level of travel on the plate. Their nature or
characters are identified by means of suitable detection
techniques. TLC System consists of a TLC plates preferably
readymade with stationary phase: These are stable and
chemically inert plates on to whose surface a thin layer of
stationary phase is applied. The stationary phase on the plates
is of uniform thickness and consists of fine particle size.
Rf value= a/b
= Distance travelled by the solute/ Distance travelled by the
solvent
a= Distance travelled by the solute
b= Distance travelled by the solvent
TLC of Hydroalcoholic extract of Pippali
A mixture of 6 ml of Toluene, 6 ml of Ethyl acetate, 1.8
formic acid, 0.25 methanol.
Heat: Heat at 110 °C for 10 minutes and examines the plate
under day light
Fig 1: Tlc Fringer Print of Hydroalcoholic Extract of Pippali
Fig 2: HPTLC (High Performance Thin Layer Chromatography)
Methodology
0.3g of extract was dissolved with 1 ml of water and 1ml
of ethyle alcohol and 3, 6 and 9µl of the above extract
was applied on a pre-coated silica gel F254 on aluminum
plates to a band width of 7 mm using Linomat 5 TLC
applicator.
The plate was developed in Toluene: Ethyl-acetate:
formic acid: methanol (6:6:1.8:0.25). The developed
plates were visualized in UV 254 and 366 and scanned
under UV 254 and 366 nm. Rf, of the spots and
densitometric scan were recorded.
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Fig 3: HPTLC chromatogram of hydroalcoholic extract of Pippali (Fruit)
Table 3: HPTLC chromatogram of Hydroalcoholic extract of Pippali
Peak
Start Rf
Start Height
Max Rf
Max Height
Max %
End Rf
End Height
Area
Area %
1
-0.04
6.1
-0.03
40.7
1.30
-0.02
0.5
427.4
0.32
2
-0.01
1.5
0.04
738.3
23.61
0.08
184.3
19224.6
14.49
3
0.08
186.0
0.09
273.6
8.75
0.11
128.5
5149.0
3.88
4
0.11
129.5
0.13
174.6
5.56
0.16
74.5
4207.4
3.17
5
0.16
74.5
0.20
107.1
3.44
0.24
56.1
4856.6
3.66
6
0.26
60.1
0.28
76.1
2.43
0.29
71.5
1805.7
1.36
7
0.30
75.1
0.33
93.8
3.00
0.35
84.1
3425.3
2.56
8
0.35
84.2
0.38
117.3
3.75
0.41
97.7
4566.8
3.44
9
0.42
96.7
0.44
100.7
3.22
0.57
83.6
3571.5
2.69
10
0.48
84.0
0.49
95.3
3.05
0.51
86.6
2059.5
1.55
11
0.51
86.9
0.52
98.5
3.15
0.55
39.1
2217.6
1.67
12
0.57
38.8
0.68
693.4
22.17
0.79
163.5
61664.4
46.48
13
0.79
164.0
0.83
397.8
12.72
0.88
93.1
16281.7
12.27
14
0.88
93.3
0.90
120.1
3.84
0.93
1.5
3217.1
2.42
Discussion and Conclusion
Pharmacognosy may be defined as a branch of bioscience
which treats in detail medicinal and related product of crude
or primary type obtained from plant, animal and mineral
origins. In short it is an objective study of crude drugs from
natural sources treated scientifically and it encompasses the
knowledge of the history, distribution, cultivation, collection,
processing for market and preservation, the study of sensory,
physical, chemical, and structural characters and the uses of
crude drugs [10].
The macroscopic and microscopic identifying characters of
useful part of Pippali was included from the Ayurvedic
Pharmacopoeia of India monograph considering the legal
document of the Government of India. Powder microscopy of
Fruit showed the presence of Fragments of thin walled cell,
starch grains, Lignified sclerenchymatous cells, calcium
oxalate crystals, Aleurone grains and oil globules. On
physiochemical analysis, the moisture content was found in-
Pippali 9.34%. The Total ash was found 4.4%; Acid insoluble
ash was 0.05%, and; water soluble ash was 2.077% in fruit of
Pippali. The phytochemical investigation shows the presence
of alkaloid, sterol and protein in fruit of Pippali. The TLC and
HPTLC was performed and the developed plates were
visualized in UV 254nm, 366nm, which are presented in
previous picture as TLC finger print as well as HPTLC
chromatogram of hydro- alcoholic extract of Pippali. This
study will help both physicians and researchers in their
respective fields, either for the treatment of diseases or in the
field of research.
References
1. The Ayurvedic Pharmacopoeia of India, First edition,
New Delhi; Govt. of India, ‘Ministry of Health and
Family welfare’ Department of Indian Systems of
Medicine and Homeopathy, 1999, 133-134.
2. Database on Medicinal Plants Used in Ayurveda, by PC
Sharma, MB Yelne and TJ Dennis, Central Council For
Reseach In Ayurveda & Siddha, Reprint Janakpuri, New
Delhi, Part I, 2005; II(3):472
3. Bhavaprakasha Nighantu of Bhavamishra, Commentary
by Krishnachandra Chunekar, Edited by GangÁsahÁya
Pandey, Chaukambha Bharati Academy, Varanasi,
Reprint, 1999, 15.
4. Caraka Samhita. of Agnivesa, elaborated by Caraka and
Dridabala, Edited with ‘Caraka-Candrika’ Hindi
commentary along with special deliberation by Dr.
Brahmanand Tripathi, Chaukambha Surbharati
Prakashan, Varanasi, 3rd Edition, 1994.
5. Sushruta Samhita. Edited with Ayurveda Tattva-
Sandipika by Kaviraja Ambika Dutta Shastri;
Chaukambha Sanskrit Sansthan, Varanasi, 5th edition,
1982
6. Ashanga. Samgraha of Vagbhata with Indu Vyakhya by
Prof. Jyotir Mitra, Chaukambha Sanskrit Series,
Varanasi, 13th Edition, 2008.
7. Ashtanga Hridaya of Vagbhata, Translated by Prof. K.R.
Srikantha Murthy, Krishnadas Academy, Varanasi, 3rd
Edition, 2000.
8. Dravyaguna Vigyana by Dr. Gyanendra Pandey,
Chaukhambha Krishanadas Academy, Varanasi, Reprint
2004; 3:116-122.
9. Dravya Guna Vigyana by P.V. Sharma Chaukhambha
Bharati Academy, Varanasi, Reprint 2003; II:278.
10. Kokate CK, Purohit AP, Gokhale SB. Pharmacognosy,
Fiftieth Edition, Nirali Prakashan, Shivaji Nagar, Pune,
2014, 1.3
