Publication Ref No.

: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-9/NOV/006 ISSN 0974 – 9446

PHARMACOGNOSTIC EVALUATION AND PHYTOCHEMICAL ANALYSIS OF THE TUBER OF

AMORPHOPHALLUS PAEONIIFOLIUS

Yadu Nandan Dey*1, Ajoy Kumar Ghosh2

1
Central Council for research in Ayurveda and Siddha,
Janakpuri, New Delhi, India
1
Gupta college of Technological Sciences,
Ashram More, Asansol, West Bengal, India
Yadu Nandan Dey Email:yadunandan132@gmail.com

ABSTRACT

The present research work discusses the development of a UV spectrophotometric method for
Metronidazole. Simple, accurate and cost efficient spectrophotometric method has been developed for
the estimation of Metronidazole (MND) in Tablet dosage form. The optimum conditions for the analysis
of the drug were established. The maximum wavelength (λ max) was found to be 277nm. The
percentage recovery of Metronidazole was in the 98.2±0.129. Beers law was obeyed in the
concentration range of 1-10µg/ml. Calibration curves shows a linear relationship between the
absorbance and concentration. The line equation y=0.0283x+0.0407 with r2 of 0.9902 was obtained.
Validation was performed as ICH guidelines for Linearity, accuracy, precision, LOD and LOQ. The
sample solution was stable up to 36 hours. The proposed method may be suitable for the analysis of
MND in tablet formulation for quality control purposes.

Key Words: Phytochemical analysis, Amorphophallus paeoniifolius, Ash value, Extractive values,
Physical constants

INTRODUCTION modern drugs are natural product origin and

In recent years, the use of herbal medicines
worldwide has provided an excellent opportunity
to India to look for therapeutic lead compounds
from our ancient system of therapy, i.e.
Ayurveda, which can be utilized for
development of new drug. Over 50% of all
they play an important role in drug development
programs of the pharmaceutical industry1.
Ash values are used to determine quality and
purity of a crude drug. Ash contains inorganic
radicals like phosphates, carbonates, silicates
of sodium, potassium, magnesium, calcium etc.
Sometimes, inorganic variables like calcium

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Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-9/NOV/006
oxalate, silica, carbonate content of the crude
drug affects ‘Total ash value’. Such variables
are then removed by treating with acid like
Hydrochloric acid and then acid insoluble ash
value is determined. Extractive values are
useful for the evaluation of the drug. It gives
idea about the nature of the chemical
constituents present in a crude drug. It is useful
for the estimation of specific constituents,
soluble in that particular solvent used for
extraction. Loss on drying is the loss in weight
in % w/w resulting from water and volatile
matter of any kind that can be driven off under
specified conditions.2,3 The tubers of wild plants
are highly acrid and cause irritation in throat and
mouth due to excessive amount of calcium
oxalate present in the tubers. The tubers are
anodyne, antiinflammatory, antihaemorrhoidal,
haemostatic, expectorant, carminative,
digestive, appetizer, stomachic, anthelmintic,
liver tonic, aphrodisiac, emmenagogue,
rejuvenating and tonic. They are traditionally
used in arthralgia, elephantiasis, tumors,
inflammations, hemorrhoids, hemorrhages,
vomiting, cough, bronchitis, asthma, anorexia,
dyspepsia, flatulence, colic, constipation,
helminthiasis hepatopathy, splenopathy,
amenorrhea, dysmenorrhoea, seminal
weakness, fatigue, anemia and general
debility4. The tuber is reported to have
antiprotease activity5, analgesic activity6,
cytotoxic activity7 , anti-inflammatory activity8
and CNS depressants activity.9
In view of this, an attempt has been made to
study the pharmacognostic evaluation and
phyto-chemical analysis of the different extacts
of the tuber of Amorphophallus paeoniifolius.
The objective of the present research is to
prove the traditional uses of the plant A.
paeoniifolius due to its active bioactive
phytocontituents. Further it is planned to isolate
the active phytoconstituents by using
biologically action guided extraction procedure.
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MATERIALS AND METHODS
Plant
The tuber of Amorphophallus paeoniifolius
(Dennst.) Nicolson var.campanulatus (Decne)
(Areaceae), was collected from Asansol, West
Bengal, India. The tuber of Amorphophallus
paeoniifolius (Dennst.) Nicolson
var.campanulatus (Decne) Sivadasan
(Areaceae), was
identified with the Herbarium of Botanical
Survey of India, Botanic Garden, Howrah with
ref no. CNH/I-I/ (272)/ 2008/ Tech. II/ 314.
Preparation of extract
The tuber of the plant was dried in shade and
made to fine powder using a laboratory mill. The
dried powder is stored in dessicator. The dry
powder is extracted with methanol and
petroleum ether using soxhlet extractor.
PHARMACOGNOSTIC EVALUATION OF THE
PLANT
A. Ash value and extractive values
The dried powder of the tubers of the plant were
taken and the parameters like moisture content,
total ash, acid-insoluble ash,
alcohol(90%ethanol) and water soluble
extractive values were determined.10
B. Acid neutralizing capacity
The Acid-Neutralizing capacity was carried out
as per USP 31. All tests were conducted at
temperature 37± 3°C. A pH meter was
standardized using the 0.05M potassium
biphthalate buffer. Magnetic stirrer was used to
produce stirring rate 300± 30 rpm. 0.5 g of ash
of tuber of Amorphophallus paeoniifolius was
transferred to 250 ml beaker and 70 ml distilled
water was added. It was mixed with magnetic
stirrer for 1 min. Then 30 ml 1.0 N Hcl was
added to the test solution with continuous
stirring for 15 min. Excess Hcl was titrated with
0.5 N NaOH to attain a stable pH of 3.5. The
number of mEq of acid consumed was
calculated by formula:
Total mEq = (30×NHcl) – (VNaOH×NNaOH)
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Where NHcl and NNaOH are normalities of
hydrochloric acid and sodium hydroxide
respectively and VNaOH is volume of sodium
hydroxide and the result were expressed as
total mEq per gm of substance.11
C. Loss on drying
The loss on drying procedure was carried
out as per Indian Pharmacopoeia (1996)
Method A. Weigh a glass-stoppered, shallow
weighing bottle.Transfer to the bottle 1 g of the
sample, cover it and accurately weigh the bottle
and the contents. Distribute the sample as
evenly as practicable by gentle sideways
shaking to a depth not exceeding 10 mm.Place
the loaded bottle in a hot air oven at 105 °C for
1 hour with the stopper remains open. After 1
hour, close the stopper and allow it to cool to
room temperature in desiccators before
weighing. Weigh the bottle and the contents.
D. Morphology of the Plant:
A stout herbaceous plant with underground
hemispherical depressed dark brown corm;
leaves compound,large,solitary, petiole stout,
mottled, 60-90 cm long, leaflets 5-12.5 cm long
of variable width, obovate or oblong, acute,
strongly and many nerved; male and female
inflorescences contiguous, neuters absent,
appendage of spadix sub-globose or
amorphous,equally or longer than the fertile
region, spathe campanulate, pointed
,strongly,closely veined, greenish-pink
externally,base within purple,margins
recurved,undulate and crisped, male
inflorescence subturbinate, female 7.5 cm or
more long; fruits obovoid 2-3 seeded red
berries.12,13,
PHYTOCHEMICAL TESTS
The qualitative chemical tests for various
phytoconstituents were carried out for
methanolic and petroleum ether extracts of
Amorphophallus paeoniifolius.The phyto
chemical tests were performed for the presence
of Alkaloids (Mayer’s test, Dragendorff’s test,
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ISSN 0974 – 9446
Wagner's test, Tannic acid test),
Glycosides(Free sugar, Raymond’s test, Legal’s
test, Bromine water Test), specific glycosides
like Saponin Glycosides(Froth Test, Hemolysis
test), Anthrauinone Glycosides(Borntrager's
test, Modified Borntrager's test), Cardiac
Glycosides(Kedde’s test, Keller killiani test),
Cyanogenetic Glycosides, tannins & Phenolic
Compounds(Gelatin test, Ferric chloride test,
Vanillin Hydrochloride test, Alkaline reagent
test, Mitchell’s test), Flavonoids (Shinoda test,
Zinc Hydrochloride reduction test, Alkaline
reagent test), Proteins & Amino Acids(Millons
test, Ninhydrin test), Sterols & Triterpenoids
(.Libermann-Buchard test, Salkowski test),
Carbohydrates (Molisch's test, Benedict's test,
Barfoed's test, Camnelisation, Selwinoff’s test,
Tollen's test, Bromine water test, Fehling's test),
Fats & Fixed Oils (Stain test, Saponification
test)14,15,16,17,18
RESULTS
( Refer Table No. 01, 02 and 03 )
Table 1 The physical properties of the extracts
of the tuber
Table 2 The Pharmacognostic parameters of
the extracts of the tuber
Table 3 Qualitative Phyto-Chemical Test
Phytochemical screening of methanolioc extract
indicated the presence of steroids, flavonoids,
alkaloids and carbohydrates.
Phytochemical screening of Petroleum ether
extract indicates the presence of steroids, fats
and fixed oil.
DISCUSSION:
From the above study it was found that the ash
value of the dried tuber was found to be 7.35%.
The water soluble ash and the acid insoluble
ash were 3.09 and 1.47 respectively. The acid
neutralizing capacity of the plant was 12.066
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 Publication Ref No.: IJPRD/2010/PUB/ARTI/VOV-2/ISSUE-9/NOV/006
mEq of 1 N Hcl and the loss on drying was
found to be 5.1%. Phytochemical screening of
methanolioc extract indicated the presence of
steroids, flavonoids, alkaloids and
carbohydrates.Phytochemical screening of
Petroleum ether extract indicates the presence
of steroids, fats and fixed oil.The methanolic
extract was found to be reddish in colour having
characteristic odor viscous consistency while
the petroleum ether extract was found to be
pale yellow in color havind viscous consistency.
The percentage yield of the methanolic and
petroleum ether extracts were found to be
9.81% and 6.23% respectively.
REFERENCES:
1. Baker JT, Borris RP, Carte B, Cordell GA,
Soejarto DD. Natural product drug discovery
and development: New perspective on
international collaboration. J. Natl. Prod. 1995;
58,1325-1357.
2. Kokate CK, Purohit AP, Gokhale SB.5
Pharmacognosy. 32nd ed. Nirali Prakashan;
2005
3. Evans WC, Trease GE. Trease and Evans
pharmacognosy. W.B. Saunders Ltd. London.
2002. Pp 32, 33, 95-99, 512, 547
4. Nair RV. Indian Medicinal Plants 3, Madras:
Orient Longman; 1993. p. 118-122.
5. Prathibha S, Nambisan B, Leelamma S. Plant
Foods. Hum Nutr, 48: 247, (1995).
6. Shilpi JA, Ray PK, Sardar MM, Uddin SJ.
Analgesic activity of Amorphophallus
campanulatus tuber. Fitoterapia 2005;76:367-9.
7. Angayarkanni J, Ramkumar KM, Poornima T,
Priyadarshini U. Cytotoxic activity of
Amorphophallus paeniifolius tuber extracts in
vitro. American-Euresian J Agric Environ Sci
2007;2:395-8.
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ISSN 0974 – 9446
8. Dey YN, De S, Ghosh AK, Anti-inflammatory
activity of methanolic extracts of
Amorphophallus paeoniifolius and its possible
mechanism. International J. of Pharma and
Biosciences 2010;1(3),1-8
9. Dey YN, Das SS, Sen M, De S and Ghosh AK.
Effects of Petroleum Ether Extract of
Amorphophallus paeoniifolius Tuber on Central
Nervous System in Mice. Indian J. Pharm. Sci.,
2009, 71 (6):651-655
10. British Pharmacopoeia 11: Ash values, Acid
insoluble Ash, Water soluble and alcohol
soluble extractive. Appendix XI. (1980). Her
Majesty’s Stationery office, London. A108,
A113
11. Khandelwal KK. Practical pharmacognosy;
Techniques and experiments. 7th ed.: Nirali
prakashan;2008
12. Nair RV. Indian Medicinal Plants 3, Madras:
Orient Longman; 1993. p. 118-122.
13. Agrawal DP. Complementary and alternative
medicine: an overview. Curr. Sci. 2002; 82, 518-
524.
14. Kokate CK, Purohit AP, Gokhale SB.5
Pharmacognosy. 32nd ed. Nirali Prakashan;
2005
15. Evans WC, Trease GE. Trease and Evans
pharmacognosy. W.B. Saunders Ltd. London.
2002. Pp 32, 33, 95-99, 512, 547
16. Brain KR and Turner TD. Practical evaluation of
phytopharmaceuticals. Wright- Scientechnica,
Bristol. 1st Ed. 1975.144.
17. Ciulei I. Methodology for analysis of vegetable
drugs. UNIDO Romania. 1981. 17-25.
18. Harborne JB. Phytochemical methods. A guide
to modern technique of plant analysis.
Chapman and Hill, London. 1992.279
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TABLES AND FIGURES:

Table 1 The physical properties of the extracts of the tuber

Sl. Physical Methanolic Extract Petroleum ether

No. Characteristics Extract

1. Percentage yeild 9.81% 6.23%

2. Color Reddish Pale yellow
3. Odor Characteristic Characteristic
4. Consistency Viscous Viscous

Table 2 The Pharmacognostic parameters of the extracts of the tuber

Sl. No. Physical Constants Value

1. Ash Value 7.35 %

2. Water Soluble Ash 3.09 %
3. Acid Insoluble Ash 1.47 %
4. Acid Neutralizing Capacity 12.066±0.8 mEq of 1 N Hcl
5. Loss on drying 5.1 %

Table 3 Qualitative Phyto-Chemical Test

Methanolic Petroleum ether

Phytochemical
Extract Extract
Mayer’s Test +ve –ve
Dragendorff’s Test +ve –ve
Alkaloids Wagner’s Test +ve –ve
Hager’s Test +ve –ve
Tannic Acid Test +ve –ve
Raymond’s Test NA NA
Glycosides Legal’s Test –ve –ve
Bromine Water Test –ve –ve
Froth Test –ve –ve
Saponin Glycosides
Hemolysis Test –ve –ve

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Borntrager’s Test –ve –ve

Anthraquinone Test Modified Borntrager’s
–ve –ve
Test
Kedde’s Test –ve –ve
Cardiac Glycosides
Keller Killiani Test –ve –ve
Cyanogenetic Glycosides –ve –ve
Gelatin Test –ve –ve
Ferric Chloride Test –ve –ve
Tannins and Phenolic
Vanillin Hydrochloride
Compounds NA NA
Test
Alkaline Reagent Test –ve –ve
Shinoda Test +ve –ve
Zinc Hydrochloride
Flavonoids +ve –ve
Reduction Test
Alkaline Reagent Test +ve –ve
Proteins and Amino Millon’s Test –ve –ve
Acids Ninhydrin Test –ve –ve
Libermann-Buchard
Steroids and +ve +ve
Test
Triterpenoids
Salkowski Test +ve +ve
Molisch’s Test +ve –ve
Benedict’s Test +ve –ve
Carbohydrates
Tollen’s Test +ve –ve
Fehling’s Test +ve –ve
Stain Test –ve +ve
Fats and Fixed Oils
Saponification Test –ve +ve
+ve- Presence of phytocontituents confirmed by the phytochemical test
-ve- Absence of phytoconstituents by the phytochemical test

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