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© 2012 Faiyaz Ahmad, Izharul Hasan, Danish Kamal Chishti & Haqeeq Ahmad. This is a research/review paper, distributed under
the terms of the Creative Commons Attribution-Noncommercial 3.0 Unported License http://creativecommons.org/licenses/by-
nc/3.0/), permitting all non commercial use, distribution, and reproduction in any medium, provided the original work is properly
cited.
Antibacterial Activity of Raphanus Sativus Linn.
Seed Extract
Faiyaz Ahmad α, Izharul Hasan σ, Danish Kamal Chishti ρ & Haqeeq Ahmad Ѡ
Abstract - Raphanus sativus Linn. (Radish) is an annual herb research to better understanding of the genetic
of family Cruciferae or Brassicaceae and grown as an edible mechanism of resistance and to continue study to
root. develop new drugs either synthetic or natural (2).
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Objectives : The aim of the study is to test the potentiality of For along period of time, plants have been a
different solvent extracts (Ethanol, Methanol, Ethyl Acetate, valuable source of natural products for maintaining
Chloroform, Benzene, Aqueous hot and Aqueous cold) human health (3). Plants are used medicinally in
against various pathogenic bacterial strains E.coli (ATCC-
different countries and are a source of many potent and
25922), Klebsiella pneumonia (ATCC-27736), Proteus vulgaris
powerful drugs. Medicinal plants represent a rich source 25
(ATCC-6380), Pseudomonas aeruginosa (ATCC-27853),
Staphalococcus aureus (ATCC-25923), Shigella sonnie of antimicrobial agent (4). Different parts of plants, herbs
(ATCC-25931), Salmonella typhi (ATCC-25241) and and spices have been used for many years for the
A
temperate and warm countries (8). It is well reputed in
ccording to World Health Organization (WHO), the Unani System of Medicine, useful for urinary complaints
increase of resistance to antibiotics by bacterial and piles. Almost all parts of the plant including leaves
pathogens is a growing problem in both seeds and roots are utilized in medicine. The fresh
developed and developing countries (1). The problem of juices obtained from leaves are diuretic, laxative. Roots
microbial resistance is growing and the outlook of the are used for urinary complaints and syphilitic disease;
use of antimicrobial drugs in future is uncertain. they are a reputed medicine for piles and gastrodynic
Therefore action must be taken to reduce this problem, pains. The seeds are expectorant, diuretic, laxative,
for example, to control the use of antibiotics, to develop carminative, antitussive and stomach tonic (8, 9, 10, and
11). The present study aims at assessing the
Author α : Dept. Of Ilmul Advia (Pharmacology), Govt. Nizamia Tibbi
antibacterial property of R. sativus seed extract, to
College, Hyderabad, Andhra Pradesh. substantiate the use of radish in Unani System of
Author σ : Lecturer, Dept of Juris and Toxicolgy, AU Tibbia College, medicine in infectious diseases.
Karolbagh, New Delhi. E-mail : izhunaz@gmail.com
Author ρ : Dept of physiology, AU Tibbia College, Karolbagh, New
Delhi.
Author Ѡ : Dept of Ilmul Advia (Pharmacolgy), NIUM Bangalore.
II. Materials and Methods plates. 3-4 colonies should be selected from the agar
plate culture. The top of the each colony is touched with
a) Plant Materials loop & transferred in to into a test tube containing4-5 ml
The sample of seeds of Raphanus sativus nutrient broth. The test tubes which containing broth
[Tukhm-e-Mooli] were collected from local market of cultured are incubated at 37◦C until it achieves the
Hyderabad, Andhra Pradesh, and was properly identi- turbidity.
fied authenticated on the basis of literary description
available in the Unani classic as well as modern g) Evaluation of Antibacterial Activity
literature by Botanist Dr. V.C. Gupta, Deputy director, The in-vitro antibacterial activity of the extracts
Central Research Institute Of Unani Medicine, was determined by agar well diffusion assay (Reeves,
Hyderabad (C.R.I.U.M.) and Dr. Hakeem. Mohd 1989). All strains were first grown in Mueller Hinton broth
Yadullah Ex. C.M.O. Govt. Nizamia General Hospital (MHB) under shaking condition for 4 h 370C and after
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Hyderabad and renowned Unani practitioner. Voucher the incubation period 0.1ml of the test inoculums was
sample was prepared and preserved in the Herbarium spread evenly with a sterile glass spreader on Mueller
of C.R.I.U.M., Hyderabad for further reference. Hinton Agar (MHA) plates. The seeded plates were
allowed to dry in the incubator at 370C. Wells were made
b) Preparation of plant Extract using sterile 6 mm cork borer in the inoculated MHA
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Different extracts of Raphanus sativus seeds plates. The wells were filled with 200µl of the extracts
were prepared for analysis in the present study (a) (re-suspended in respective solvents) and negative
Ethanol (b) Methanol (c) Ethyl Acetate (d) Chloroform (e)
Global Journal of Medical Research Volume XII Issue XI Version I
compared with standard antibiotics Chloramphenicol present in the plants are known to be toxic to micro-
(25µg) and Ciprofloxacin (25µg). organism (24). Antibacterial activity of tannins may be
The plant extracts were also screened for related to their ability to inactivate microbial adhesins,
qualitative analysis to know the relative distribution of the enzymes and cell envelope transport proteins, they also
secondary metabolites which may be responsible for the complex with polysaccharides (25) .The broad spectrum
potent antibacterial activity. Flavonoids are extracted antibacterial activity exhibited by R. sativus may be
into Ethanol, Aqueous hot and Aqueous cold. Alkaloids attributed to the various active constituents presents in it
are extracted into Ethanol, Methanol, Chloroform, which either due to their individual or combined action.
Aqueous hot and Aqueous cold. Glycosides are present Thus, the study ascertains the value of R. sativus used
in all solvents. Carbohydrates are extracted only in in Unani System of Medicine. This could be of
Methanol, Aqueous hot and Aqueous cold. Phenols are considerable interest to the development of new drugs.
extracted into Ethanol, Chloroform and Aqueous hot.
Saponins are extracted into Methanol, Chloroform, V. Acknowledgments
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Aqueous hot and Aqueous cold. Sterols are found in The authors are grateful to Dr. Mushtaq Ahmad,
Ethanol, Methanol, Chloroform and Benzene. Tannins Director Central Research Institute of Unani Medicine
are extracted into Ethanol, Chloroform and Aqueous hot. (CRIUM) Hyderabad for the provision of laboratory
While Protiens are present only into Benzene. Table3. space for the extraction process and equipments to 27
carry out this research, as well as for his suggestions
IV. Discussion and encouragement.
11. Chopra, R. N., S. L. Nayar, and I. C. Chopra. 1986 phytochemical studies on twelve species of Indian
Glossary of Indian Medicinal plants (Including the medicinal plants. African Journal of Biology Res.,
supplements) Council of Scientific and Industrial 10: 175-181.
Research, New Delhi. 19. Aliero, A. A. and A. J. Afolayan, 2006. Antimicrobial
12. Reeves, D. S., 1989. Antibiotic assays, In: Howkey. activity of Solanum tomentosum. African Journal of
P. M., Levis, D. A. (Eds.), Medical Bacteriology, A Biotechnology, 5: 369-372.
practical Approach. IRL Press, Oxford, pp.195- 20. Samy, R. P. and S. Ignacimuthu, 2000. Antibacterial
221(chapter). activity of some folklore medicinal plants used by
13. Forbes BA, Sahm OF, Weissfeld AS and Trevomp tribals in Western Ghats in India. J.
EA. Methods for testing antimicrobial effectiveness. Ethnopharmacol., 69: 63-71.
Bailey and Scott’s Diagnostic Microbiology MOS by 21. Behera, S. K. and M. K. Misra, 2005. Indigenous
Co: St Louis, Missouri, PD 1990: 171-194. phytotherapy for genito-urinary diseases used by
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modern techniques of plant analysis, 3rd Edn. 24. Mason, T.L. and Wasserman, B.P Phytochem 1987;
Chapman and Hall London 1998: 302-312. 26: 2197-2202.
17. Tona, L., K. Kambu, N. Ngimbi, K. Cimanga and A. 25. Ya, C., Gaffney, S.H., Lilley, T.H. and Haslam, E.
J. Vlietinck, 1998. Antiamoebic and phytochemical Carbohydrate polyphenol complexation. In
screening of some Congolese medicinal plants. J. Hemingway, R.W. and Karchesy J.J., Eds.,
Ethnopharmacol., 61: 57-65. Chemistry and Significance of Condensed Tannins,
18. Parekh, J. and S. Chanda, 2007b. Antibacterial and Plenum Press, New York. 1998: 552-553.
Table : 1
3. Glycosides
(a) Conc.H2 SO 4 1ml of conc.H2SO 4 was Formation of Present
test added to 1ml of test reddish colour
solution and is allowed
to stand for 2 minutes.
(b) Aq NaOH test To alc. Ext. of the drug Formation of
add 1ml of water and yellow colour Present
adds aq.NaOH
solution.
4. Carbohydrates
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(a) Benedict’ s test To o.5 ml of aq. Ext. Formation of Present
of the drug add 5 ml of colour ppt.
Benedict ’ s solution and
boil for 5 min. A red - violet
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(b)Molisch ’ s test To 2 ml of aq. Ext. of ring is formed Present
the drug add 2 drops of at the junction
8. Tannins
Ferric chloride test To 1- 2 ml of aq. ext. of A bluish black Present
the drug add few drops of colour is
5% aq. ferric chloride produced
solution. which
disappear on
addition of a
few ml of a dil.
Sulphuric acid
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solu tion
followed by
the formation
of a yellow-
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2 brown ppt.
9. Proteins
Global Journal of Medical Research Volume XII Issue XI Version I
EXTRACTS
E.coli K.pneumoniea P.valgaris Ps.aeruginosa S.aureus S.sonnie S.paratyphi S.typhi
CIPROFLOXACIN
(25µG) 27±0.4 26±0.3 30±0.4 30±0.4 25±0.5 27±0.4 30±0.5 35±0.5
Table : 3
Secondary Metabolites Name of Test Results (+/-)
Et Mt Ea Ch Ben Aq H Aq C
Alkaloids Dragendroff’s ++ ++ -- ++ -- ++ ++
Flavonoids Shinoda ++ -- -- -- -- ++ ++
NaOH ++ -- -- -- -- ++ ++
Glycosides Conc.H2SO4 ++ ++ ++ ++ ++ ++ ++
Aq NaOH ++ ++ ++ ++ ++ ++ ++
Carbohydrates Benedict’s -- ++ -- -- -- ++ ++
Molisch’s -- ++ -- -- -- ++ ++
Phenol Ferric chloride ++ -- -- ++ -- ++ --
Aq. lead acetate ++ -- -- ++ -- ++ --
Saponins Foam -- ++ -- ++ -- ++ ++
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Sterols Salkowski ++ ++ -- ++ ++ -- --
Tannins Ferric chloride ++ -- -- ++ -- ++ --
Proteins Millon’s -- -- -- -- ++ -- --
Et= Ethanol, Mt= Methanol, Ea= Ethyl acetate, Ch= Chloroform, Ben= Benzene, Aq=Aqueous, H=Hot, . 31
C=Cold
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