‘

PROJECT WORK SUBMITTED TO ASSAM SCIENCE AND TECHNOLOGY

UNIVERSITY, GUWAHATI, ASSAM FOR THE PARTIAL FULFILLMENT OF
THE REQUIREMENT FOR THE AWARD OF DEGREE OF

BACHELOR OF PHARMACY

Under the guidance of Mr. Biswajit Dash (M.Pharm)

Assistant Professor, Dept. of Pharmaceutical Chemistry (GIPS)

SUBMITTED BY
UTTAM BORAH
Regd. No. 097805514 of 2014-15
Roll No. 1405511005
Sub Code- PY1327210

GIRIJANANDA CHOWDHURY INSTITUTE OF

PHARMACEUTICAL SCIENCES, HATHKHOWAPARA, AZARA,
GUWAHATI-781017 KAMRUP, ASSAM (INDIA)
PREFACE

Pharmacy is a profession which is concerned with the art and science of suitable and
convenient material for distribution and use in the treatment and prevention of disease, so it
is a fully technical profession where practical knowledge is much more important along with
theoretical knowledge. So being a Pharmacy student it is my pleasure and privilege to submit
this dissertation in its present form. The progress of scientific research has been increasing
sustainability in the last two decades particularly in the field of pharmacy. The ultimate
biochemical objective of all these are remain same, to develop better drugs for a specific
disease. Here I have tried to find out the protective effects of antimicrobial drugs.

In this literature review I have written an introduction part where the details about the
herbal drug with its advantage, disadvantage, and definition of antimicrobial activity is
summarized. In this literature review I have shown the reason why I have performed this
project work. The possible literature regarding Leucas plukenetii (Roth)Spreng are written
in literature review section. The reference of my work is given separately.

At last I expect this assignment can give some guidelines about further Phytochemical
and Pharmacological research.

UTTAM BORAH
7th SEMESTER
Bachelor of Pharmacy (GIPS)
CERTIFICATE

This is to certify that the topic incorporated in this Project work entitled
“PRELIMINARY PHYTOCHEMICAL SCREENING & ANTIMICROBIAL
ACTIVITY OF ETHANOLIC EXTRACT OF WHOLE AERIAL PART OF THE
HERB Leucas plukenetii (Roth) Spreng” being submitted by UTTAM BORAH Roll No-
1405511005 Reg. No- 097805514 in the partial fulfilment of the requirement of the award
of degree of Bachelor in Pharmacy (B. Pharm) of Girijananda Chowdhury Institute of
Pharmaceutical Science, Guwahati, Assam is a benefited assignment which has been
carried out my direct supervision and guidance during the academic Session 2016-17.

Supervised by Verified By
Mr Biswajit Dash (Principal, GIPS)
Asstt. Professor, GIPS
ACKNOWLEDGEMENT
“Karma-yoga is a supreme secret indeed”
- Srimadbhagwadgita

I would first like to thank to ‘LORD NARAYANA’ & “GURU MADHABDEV”

for who’s blessing only this literature review cum research work was possible.

I consider it a great privilege & honor to have had the opportunity to undergo the
Project work in Girijananda Chowdhury Institute of Pharmaceutical Science, Guwahati,
Assam. Hence, I would like to offer my heartiest thanks to Prof (Dr). Suvakanta Dash,
Principal of GIPS.

I am greatly indebted to Mr Biswajit Dash ; Assistant Professor, Department of

Pharmaceutical Chemistry, GIPS, for his valuable guide and suggestion.

I owe a dept of gratitude to Dr P.P. Baruah , HOD; Department of Botany, Guwahati

University for authenticate my specimen.

I convey my heartiest thanks to Dr Damiki Laloo , Assistant Professor, Department

of Pharmacognosy, GIPS for most valuable suggestions, constant encouragement, and
affectionate guidance in and Mr. Diganta Das (Librarian), GIPS for their support for
carrying out this projects.

At last, I am greatly thankful to my friends in GIPS for extending their constant

cooperation which went a long way towards the completion of this Literature review cum
research work.

UTTAM BORAH
7th SEMESTER
Bachelor of Pharmacy (GIPS)
DEDICATED TO MY BELOVED
PARENTS….
CONTENTS

1. AIM AND OBJECTIVES 1-2

2. INTRODUCTION 3-6

3. LITERATURE REVIEW 7-12

4. PLAN OF WORK 13-15

5. BIBLIOGRAPHY 16-19
CHAPTER-1

AIM & OBJECTIVES

Page 1
CHAPTER 1 AIM & OBJECTIVES 2016
1.1. AIM:

Antimicrobial resistance is a growing problem due to overuse of antibiotics in

humans. Many of the currently used antibacterial are associated with adverse effects such as
toxicity, hypersensitivity, immune suppression, and tissue residues posing public health
hazard. As the global scenario is now changing towards the use of non- toxic and eco-friendly
products, development of modern drugs from traditional medicinal plants should be
emphasized for the control of various human and animal diseases. In the present study, the
selection of this plant for evaluation was based on its traditional usages.

So further new alternative remedies for treatment of bacterial diseases are usually
required. Although very few works have been done on the antimicrobial activity of his
medicinal plant, it needs further study for verification of its activity against disease causing
microorganisms. Thus the aim of this project work is the evaluation of the antimicrobial
potency of Leucas plukenetii (F-Laminaceae)

1.2. OBJECTIVES:

The present objective of the study is-

 Extraction of active constituents from the dried leaves of Leucas plukenetii

 Preliminary Phytochemical screening of the extract of leaves of Leucas plukenetii
 Antimicrobial activity of the extract in comparison to suitable standard.

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CHAPTER-2

INTRODUCTION

Page 3
CHAPTER 2 INTRODUCTION 2016
2.1. INTRODUCTION:

The medicinal use of plants is very old. The writings indicate that therapeutic use of plants
is as old as 4000–5000 B.C. and Chinese used first the natural herbal preparations as
medicines. In India, however, earliest references of use of plants as medicine appear in
Rigveda, which is said to be written between 1600-3500 B.C.

Later the properties and therapeutic uses of medicinal plants were studied in detail and
recorded empirically by the ancient physicians in ‘Ayurveda’ which is a basic foundation of
ancient medical science in India [1]. The Hindu Physician, Susruta enlisted seven hundred
and sixty medicinal plant in Ayurveda in the 5th century AD. [2]

Antibiotic resistance is a growing problem. The widespread and indiscriminate use

of antibacterial agents resulted in development of drug resistance among many virulently
pathogenic bacteria species. Many of the currently used antibacterial are associated with
adverse effects such as toxicity, hypersensitivity, immune suppression, and tissue residues
posing public health hazard.

So, further new alternative remedies for treatment of bacterial diseases are usually
required. As the global scenario is now changing towards the use of non- toxic and eco-
friendly products, development of modern drugs from traditional medicinal plants should be
emphasized for the control of various human and animal diseases. [3]

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CHAPTER 2 INTRODUCTION 2016
2.2. NATURAL SOURCES OF ANTOMICROBIAL AGENTS(AMA).

Sl Plant Scientific Name Major components

No
1 Allspice Pimenta dioica Eugenol, methyl ether cineol
2 Basil Ocimum asilicum d-linalool, methyl chavicol,
geraniol
3 Black Pepper Pipper nigrum Monoterpenes, Sesquiterpenes

4 Caraway Carum carvi Carvone, limonene

Seed
5 Celery Seed Apium graveolens d-limonene
6 Cinamon Cinamonum Cinnamic aldehyde, l-linalool,
zeylanicum p-cymene, eugenol
7 Clove Syzygium aromaticum Eugenol, cariofilene
8 Coriander Coriandum sativum d-linalool, d-pinene,
9 Cumin Cuminum cyminum Cumin aldehyde, p-cymene
10 Fennel Foeniculum vulgare Anethole
11 Garlic Allium sativum Diallyl disulphide, diethyl
sulphide, allicin
12 Lemongrass Cymbopogon citratus Citral, geraniol
13 Marjoram Origanum majorana Linalool, cineol, methyl
chavicol, eugenol, terpininecol
14 Mustard Brassica hirta Allyl-isothiocynanate
15 Onion Allium cepa d-n-propyl disulphide, methyl-n-
propyl disulphide
16 Oregano Origanum vulgare Thymol, carvacrol, p-cymene
17 Parsley Pertroselinum Pinene, fenol-eter-apiol
crispum
18 Rosemary Rosmarinus officinalis Borneol, cineol, borneol,
thymol, eugenol
19 Sage Salvia officinalis Thujone, cineol, borneol,
thymol, eugenol
20 Tarragon Artemissia Methyl chavicol, anethode
dracunculus
21 Thyme Thymus Vulgaris Thymol, carvacol, geraniol, p-
cymene
22 Vanilla Vanilla planifolia Vanillin, Vanillic, p-
hydroxybenzoic, p-cumaric
acids
Fig-1: Sources of Natural AMA [4]

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CHAPTER 2 INTRODUCTION 2016
2.3.PLANT PROFILE

2.3.1. BIOLOGICAL SOURCE: It consisted of the dried whole herb of Leucas

Plukenetii belonging to the family Lamiaceace.

2.3.2 PLANT DESCRIPTION-

 Habit: An annual herb with square stem.
 Leaves : Linear-oblong or oblong lanceolate.
 Flowers : Sessile, ciliate with long hairs.
 Fruit: Oblong, inner face sharply angular
 .Flowering and Fruiting Time : July-October
 Plant Form : Herb [5]

2.3.3 VERNICULAR NAME- Doron Bon [6]

2.3.4 SYNONYM- Leucas aspera [7]

Fig-2. Leucas plukeneti

2.3.5 ETHNOMEDICINAL USES OF LEUCAS PLUKENETII-

 The leaves and flowers are used by the Kavirajes to treat tooth infections and mucus.
[8]
 It is applied on inflamed parts to relieve pain and inflammation. [9]
 Leave paste made with lime juice applied externally in the treatment of Headache.
 Flowers were macerated and extract put dropwise into opposite side of nostril to
reduce migraine.
 Leaves with pepper and garlic chewed and spilt into the nostril with force in case of
SNAKE BITE.
 Whole plant dugout early in the morning and made into paste with water to treat
Wounds and worms. [10]

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CHAPTER-3

LITERATURE
REVIEW

Page 7
CHAPTER 3 LITEARATURE REVIEW 2016
3.1. PHYTO-CHEMICAL EVALUATION:

3.1.1. Sadhu et al. (2003) isolated eight lignans namely nectandrin B , (-)-chicanine ,
meso-dihydroguaiaretic acid, macelignan , myristargenol B , erythro-2-(4-allyl-2,6-
dimethoxyphenoxy)-1-(4-hydroxy-3-methoxy phenyl) propan-1-ol, Machilin C, (7R,8R)-
and (7S,8S)-licarin from the methanol extract of the whole plant.

3.1.2. Manivannana and Sukumar (2007) reported free flavonoid ‘baicalein’ was reported in
the ethereal fraction of hydro methanolic extract of flower.

3.1.3. Gerige et al. 2007 & Mangathayaru et al. (2006) found to contain high amount of a-
farnesene, α-thujene and menthol.

3.1.4. A new type of diterpenes, leucasperones A and B ; leucasperols A and B, have been
reported by Sadhu et al. (2006).

3.1.5. Linoleic acid were found which was contingent upon crop variation by Chen et al.,
(1979).

3.1.6. Rajyalakshmi et al., (2001) found Leucas aspera and Leucas cephalotes contain
significant amounts of total carotenoid and β-carotene. [11]

3.2. PHARMACOLOGICAL ACTIVITY:

3.2.1 Antimicrobial activity: Mangathayaru et al., (2005) reported significant antimicrobial

activity was reported for the alkaloidal fraction and the total methanol extract the
flowers [12].

Satyal et al., (2013) reveals that the essential oil of the plant showed no activity
against E. coli, P. aeruginosa, and C. albicans (MIC ≥ 1250μg/mL). The oil did exhibit good
activity against S. aureus (MIC = 625 μg/mL), B. cereus (MIC = 313μg/mL), and A. niger

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CHAPTER 3 LITEARATURE REVIEW 2016
(MIC = 313 μg/mL), most likely attributable to the sesquiterpenes present in the oil. Both
(E)-caryophyllene and α-humulene have shown antibacterial activity against B. cereus and
S. aureus, and α-humulene was antifungal to A. niger [13]

Ilango et al., (2008) evaluated and found that Ethyl acetate extract (EAE) of whole
plant exhibited moderate to significant and concentration dependent antibacterial activity
against all the tested microorganisms at the concentrations of 50, 100, 200, 300 and 400
µg/disc and comparable to the various antibiotics used for individual microorganism. This
study also reveals that EAE was found to be highly active against Staphylococcus
epidermidis and Klebsiella pneumoniae [14]

Chew et al. (2012) evaluated the antimicrobial activity of crude extracts of root,
flower, leaf having notable antibacterial activity. The root extract showed the highest mean
zone of inhibition ranging from 9.0–11.0 mm at a concentration of 100 mg/mL.[15]

Antony et al. (2013) describes green synthesis of silver nanoparticles (AgNPs)

utilizing the plant. Antimicrobial activity of the AgNPs was tested against Aeromonas
hydrophila. Catla catla, the model organism used for the experiment was divided into six
groups with 15 animals in each group. In vivo analysis of biochemical parameters and
histological architecture provided evidence for the antibacterial effect of AgNPs in the fish
model.[16]

3.2.2 Central nervous system activity: Rahman et al. (2007) found that ethanolic extract of
root showed significant peripheral antinociceptive activity at a dose of 400 mg/kg . [17]

3.2.3 Antioxidant activity: Rahman et al. (2007) reported significant activity was found in
the ethanolic extract of root (IC50 = 7.5 µg/ml) .[17]

Chew et al. (2012) evaluated antioxidant activity methanol extract of root and it
possessed antioxidant activity near the range of vitamin E and thus could be a potential rich
source of natural antioxidant.[15]

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CHAPTER 3 LITEARATURE REVIEW 2016

3.2.4 Hepatoprotective activity: According to Thenmozhi et al (2013) the Hydroalcoholic

leaf extract has found to possess hepatoprotective property by suppressing the abnormal
elevation of liver enzymes induced by lead acetate . [18]

Mangathayaru et al.,(2005) showed The cold methanolic extract of the whole plant
of was found to exhibit significant hepato protection in CCl4 induced liver damage.[19]

Banu et al. (2012) found that d-galactosamin administration induced hepatotoxicity

in rats which was manifested by increased levels of alanine aminotransferase, aspartate
aminotransferase, alkaline phosphatase, total cholesterol, triglycerides, total bilirubin and
oxidative stress. Pre-treatment with LA extract significantly protected the liver in d-
galactosamin administered rats. LA extract significantly elevated antioxidant enzymes like
superoxide dismutase, catalase, glutathione peroxidase and decreased lipid peroxidation
levels in liver. The total phenolic and flavonoid content in LA aqueous extract was found to
be 28.33 ± 0.19 gallic acid equivalents mg/g of extract and 3.96 ± 0.57 rutin equivalent mg/g
of extract, respectively. LA extract (200 and 400 mg/Kg) treatment with CCl4 decreased the
hexobarbitone-induced sleeping time in mice by 56.67 and 71.30%, respectively, which
indicated the protective effect of LA on hepatic MDMEs. Histological studies showed that
LA at 400 mg/kg attenuated the hepatocellular necrosis in d-galactosamin intoxicated
rats.[20]

3.2.5 Anti-inflammatory activity: Reddy et al. (1986) reported that whole plant extract
have anti-inflammatory activity and caused degranulation of mast cells [17].

Reddy et al. (1986) reported anti-inflammatory activity of the yellow colored

chromatographic fraction of extract was observed in the chronic and acute models of
inflammation. It was observed that, the activity was due to the inhibition of histamine and
serotonin [21].

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CHAPTER 3 LITEARATURE REVIEW 2016
Similar observations were made by Saundane et al., (2000) and concluded that, on
preliminary screening of ethanol and dis-tilled water extracts exhibited significant anti-
inflammatory activity, whereas only ethanol (95%) extract produced long term analgesia in
the experimental animals. [22]

Srinivas et al. (2000) reported that the dried leaves of for the alcoholic and aqueous
extracts of the plant possessed significant anti-inflammatory activity against carrageenan-
induced paw oedema and cotton pellet induced granuloma. [23]

Goudgaon et al. (2003) concluded that the alkaloid fraction of the crude ethanolic
extract is accountable for the antiinflammatory activity. [24]

3.2.6 Cytotoxicity: Krishnaraju et al. (2005) proves that the hydro alcoholic extract of whole
plant exhibited cytotoxicity (LC50 = 1,900 µg/ml)[16] and this activity was more in the root
extract (LC50 = 52.8µg/ml) [25].

Chew AL et al. (2012) performed brine shrimp lethality bioassay , it was evident
that the methanol root extract did not show significant toxicity. The LC50 value for 12 h and
24 h observation was 2.890 mg/mL and 1.417 mg/mL, respectively. [15]

3.2.7 Larvicidal Activity: Maheswaram et al. (2008) The findings of the present
investigation revealed that L. aspera has good larvicidal activity of the plant activity against
Culex quinquefasciatus Say. and Aedes aegypti [26]

3.2.8 Antihyperglycemic Activity: Mannan et al. (2010) performed an experiment upon

antihyperglycemic activity of plant extract at mice. The methanol extract of leaves, when
administered to glucose-challenged mice demonstrated dose-dependent and significant
antihyperglycemic activity The stem extract also demonstrated significant and dose-
dependent antihyperglycemic activity in glucose tolerance tests in mice. [27]

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CHAPTER 3 LITEARATURE REVIEW 2016
However, dose for the antihyperglycemic activity demonstrated by stem extract was
lower than that of leaf extract. At the highest dose of 400 mg extract per kg body weight,
serum glucose levels fell by 28.39% versus 34.01% obtained with the same dose of leaf
extract. To summarize, antihyperglycemic activity at a statistically significant level was
observed with leaf extract, when the extract was administered at doses of 200 mg per kg body
weight to glucose-loaded mice. [27]

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CHAPTER-4

PLAN OF WORK

Page 13
CHAPTER 4 PLAN OF WORK 2016

4. PLAN OF WORK

• Collection of Plant Material.

• Washing of plan material.
Step-I • Authentication of Plant Material by GU.

• Drying and Grinding of plant material.

Step - II

• Extraction of Plant Material (Maceration

Process).
Step-III • Preliminary phytochemical screening of extract.

• Determination of Antimicrobial Activity.

Step-IV

Fig 3-Plan of work

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CHAPTER 4 PLAN OF WORK 2016
4.1. Collection of Plant Material: The fresh whole aerial parts of the plant Leucas
plukentii was collected from various parts of the District Lakhimpur, Assam.

4.2. Washing of Plant Material: The collected plant material was washed using water
and then it is sundried to remove moisture.

4.3. Authentication of Plant Material: Herbarium sheet was prepared using the cleaned
plant material. Then it was authenticating at Department of Botany, Gauhati University.

4.4. Drying and grinding of plant material: The cleaned plant material will be dried at
600c using Tray dryer and then it will be grinded to form powder.

4.5. Extraction of Plant Material: The sieved powder drug will be extracted by
maceration process using ethanol as solvent.

4.6. Preliminary phytochemical screening of extract: The Preliminary phytochemical

screening of extract will be done by tested the extract for glycosides, alkaloids etc.

4.7. Determination of Antimicrobial Activity: The potency of extract will be

determined by agar disc diffusion method against a standard.

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CHAPTER-5

BIBLIOGRAPHY

Page 16
CHAPTER 5 BIBLIOGRAPHY 2016
BIBLIOGRAPHY

1. Prakash P and Gupta N. Therapeutic uses of Ocimum sanctum linn (tulsi) with a note
on eugenol and its pharmacological actions; a short review. Indian Journal of
Physiology and Pharmacology 2005; 49: 125–131.
2. Guthrie D.A. History of Medicine. London, Thomas Nelson and Sons Ltd : 1945;
p-33.
3. Das A. A study of antibacterial activity of ethanolic extracts and aqueous extracts of
Leucas longifolia (doron) leaves against Eschreiashia coli. International Research
Journal of Pharmacy 2012; 3: 130-132.
4. Michael P, Sofos J, and Branen A . Antimicrobials in Food, Tailor & Francis group;
Third Edition : 2005; p- 429-507.
5. Http://www.efloraofgandhinagar.in/herb/leucas-aspera [Accessed 21 October 2016]
6. Http://www.flowersofindia.net/catalog/slides/Common%20Leucas.html [Accessed
02 November 2016]
7. http://www.theplantlist.org/tpl1.1/record/kew-111857 [Accessed 02 November
2016]
8. Mohammed R, Das AK, Ariful Md., Haque M, Rownak J, Khan M, Rahman T, and
Chowdhury MH. An Ethnomedicinal Survey of Dhamrai Sub-district in Dhaka
District, Bangladesh. American-Eurasian Journal of Sustainable Agriculture 2009; 3:
881-888.
9. Rahmatullah Q, Bhatti R, and Rabia A. Ethnomedicinal uses of herbs from northern
part of nara desert,Pakistan. Pakistan Journal of Botany 2010; 42(2): 839-851.
10. Parinitha M, Harish GU, Vivek NC, Mahesh T, and Shivanna MB. Ethnobotanical
wealth of Bhadra Wild life sanctuary of Karnataka. Indian Journal of Traditional
knowledge 2004; 3(1): 37-50.
11. Singh SK, and Chowhan HS. A review of plants of genus Leucas. Journal of
Pharmacognosy and Phytotherapy 2011; 3: 13-23.
12. Mangathayaru K, Lakshmikant J, Sundar NS, Swapna R, Grace XF, and Vasantha J.
Antimicrobial activity of Leucas aspera flowers. Fitoterapia 2005; 76: 752-754.

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CHAPTER 5 BIBLIOGRAPHY 2016
13. Satyal P, Paudel P, Poudel A, and Setzer WN. Microbiological activities of volatile
constituents of Leucas aspera (Willd). Nepal Journal of Natural Pharmaceuticals
2012 ; 3 : 118-119.
14. Ilangos k. Ramya s and Gopinath G. Antibacterial activity of Leucas aspera spreng.
International Journal of Chemical Sciences 2008; 6 :526-530.
15. Chew AL , Jessica JJA and Sasidharan S. Antioxidant and antibacterial activity of
different parts of Leucas aspera. Asian Pacific Journal of Tropical Biomedicine,
2012; 2: 176-180.
16. Antonya JJ, Nivedheethaa M, Sivaa D, Pradeephaa G, Kokilavania P, Kalaiselvia S,
Sankarganesha A, Balasundaramb A, Masilamanib V, and Achiramana S.
Antimicrobial activity of Leucas aspera engineered silver nanoparticles against
Aeromonas hydrophila in infected Catla catla. Colloids and Surfaces B:
Biointerfaces 2013; 109: 20-24.
17. Rahman MS, Sadhu SK, and Hasan CM. Preliminary antinociceptive, antioxidant and
cytotoxic activities of Leucas aspera root. Fitoterapia 2007; 78: 552-555.
18. Thenmozhi M, Dhanalakshmi M, Devi KM, Sushila K, and Thenmozhi S. Evaluation
of hepatoprotective activity of Leucas aspera hydroalcoholic leaf extract during
exposure to lead acetate in male albino wistar rats. Asian Journal of Pharmaceutical
and Clinical Research 2013; 6 : 78-81.
19. Mangathayaru K, Fatima GX, Bhavani M, Meignanam E, Karna SLR and Kumar
DP. Effect of Leucas aspera on hepatotoxicity in rats. Indian Journal of
Pharmacology 2005 ; 37 : 329-330.
20. http://www.tandfonline.com/doi/abs/10.3109/13880209.2012.685130
21. Redy MK, Viswanathan S, Sambantham PT, Ramachandran S, and Kameswaran L.
Effects of Leucas aspera on experimental inflammation and mast cell degranulation.
Ancient Science of Life 1986; 5: 168-171.
22. Saha K, Mukherjee PK, Mandal SC, Saha BP, and Pal M. Antiinflammatory
evaluation of Leucas lavandulaefolia Rees Extract. Natural Product Sciences 1997;
2: 119-122.

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CHAPTER 5 BIBLIOGRAPHY 2016
23. Srinivas K, Rao MEB, and Rao SS. Anti-inflammatory activity of Heliotropium
indicum Linn. And Leucas aspera Spreng. in albino Rats. Indian Journal of
Pharmacology 2000; 32: 37-38.
24. Goudgaon NM, Basavaraj NR, and Vijayalaxmi A . Antiinflammatory activity of
different fractions of Leucas aspera spreng. Indian Journal of Pharmacology 2003;
35: 397-398.
25. Krishnaraju AV, Rao TVN, Sundararaju D, Vanisree M, Tsay HS, and Subbaraju
GV. Assessment of bioactivity of Indian medicinal plants using brine shrimp
(Artemia salina) lethality assay. International Journal of Applied Science and
Engineering 2005; 3: 125-134.
26. Maheswaran R, Sathish S, and S Ignacimuthu S. Larvicidal activity of Leucas aspera
(Willd) against the larvae of Culex quinquefasciatus Say. and Aedes aegypti L.
International Journal of Integrative Biology 2008; 2 : 214-217.
27. Mannan A, Das S, Rahman M, Jesmin J, Siddika A, Rahman M, Rahman S,
Chowdhury M and Rahmatullah M. Antihyperglycemic Activity Evaluation of
Leucas Aspera (Willd.) Link Leaf and Stem and Lannea Coromandelica (Houtt.)
Merr. Bark Extract in Mice. Advances in Natural and Applied Sciences 2010; 4(3):
385-388.

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