ANTIBACTERIAL ACTIVITY OF DHATAKI (WOODFORDIA ...

ANTIBACTERIAL ACTIVITY OF DHATAKI (WOODFORDIA

FRUTICOSA (L.) KURZ) AS PRATINIDHI DRAVYA FOR

YASHTIMADHU (GLYCYRRHIZA GLABRA LINN) WITH REFERENCE

TO VRANAHARA KARMA By

Dr. IBAMEAIMON POHTAM

Dissertation Submitted to the

RAJIV GANDHI UNIVERSITY OF HEALTH SCIENCES BENGALURU,

KARNATAKA

In partial fulfillment of the requirements for the degree of

AYURVEDA VACHASPATI

(DOCTOR OF MEDICINE – AYURVEDA)

In

DRAVYAGUNA VIJNANA

Under the Guidance of

Dr. ANURADHA KN, M.D (Ayu)

ASSOCIATE. PROFESSOR

DEPARTMENT OF DRAVYAGUNA

SDM COLLEGE OF AYURVEDA & HOSPITAL, HASSAN

CO-GUIDE

Smt. SHASHIREKHA K S, Msc, Mphil

Microbiologist

DEPARTMENT OF ROGA NIDANA EVUM VIKRITI VIGYANA

SDM COLLEGE OF AYURVEDA & HOSPITAL, HASSAN

DEPARTMENT OF DRAVYAGUNA VIJNANA SHRI DHARMASTHALA MANJUNATHESHWARA

COLLEGE OF AYURVEDA & HOSPITAL

HASSAN – 573 201

2020

I

Abbreviations

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) aspratinidhi dravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference tovranahara karma”

IX

LIST OF ABBREVIATIONS USED

AH Astanga Hridaya

AS Astanga Sangraha

API Ayurvedic

Pharmacopeia of India

BPN Bhavaprakasha

Nighantu

CS Charaka samhitha

DN Dhanwanthari Nighantu

HPTLC High- Performance

Thin Layer

Chromatography

KN Kaiyyadeva Nighantu

MPN Madanapala Nighantu

NA Adarsha Nighantu

RN Raja Nighantu

Sha.N Shaligrama Nighantu

SS Sushruta samhita

SON Sodhala Nighantu

List of tables

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhidravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference to vranaharakarma”

X

LIST OF TABLES

List of tables

Sl no Tables PageNo.

1 Synonyms of Yashtimadhu [Glycyrrhiza glabra Linn.]

Mentioned in various Samhitha, Nighantu

6

2 Vernacular names of Yashtimadhu [ Glycyrrhiza glabra

Linn.]

8

3 Classification of Yashtimadhu [Glycyrrhiza glabra Linn.] 9 - 10

4 Rasapanchaka of Yashtimadhu [Glycyrrhiza glabra Linn.] 14

5 Karma of Yashtimadhu [Glycyrrhiza glabra Linn.] 15

6 Indications of Yashtimadhu [Glycyrrhiza glabra Linn.] 15

7 Quantitative standards of Yashtimadhu [Glycyrrhiza

glabra Linn.]

17

8 Synonyms of Dhataki [ Woodfordia fruticosa (L.) Kurz]

Mentioned in various Samhitha, Nighantu

25 - 26

9 Vernacular names of Dhataki [ Woodfordia fruticosa (L.)

Kurz]

28

10 Classification of Dhataki [Woodfordia fruticosa (L.)

Kurz]

29

11 Rasapanchaka of Dhataki [Woodfordia fruticosa (L.)

Kurz]

33

12 Karma of Dhataki [Woodfordia fruticosa (L.) Kurz] 34

13 Indications of Dhataki [Woodfordia fruticosa (L.) Kurz] 35

14 Quantitative standards of Dhataki [Woodfordia fruticosa

(L.) Kurz]

36

15 Nidana of Vrana 42

List of tables


XI

16 Lakshanas of Dushta Vrana 45

17 Vrana Varna 46

18 Vrana Gandha according to Sushruta 47

19 Vrana Gandha according to Charaka 47

20 Vrana Srava according to Sushruta 48

21 Vrana Srava according to Charaka 48

22 Vrana Vedana 49

23 Types of Ulcer 53

24 Ingredients for preparation of the extracts 65

25 Showing macroscopic evaluation of root of Yashtimadhu

(Glycyrrhiza glabra Linn.)

82

26 Showing Physiochemical parameter of Yashtimadhu


83

27 Showing results of preliminary phytochemical screening of

aqueous extract of Yashtimadhu (Glycyrrhiza glabra

Linn.)

84

28 Rf values of sample of Yashtimadhu (Glycyrrhiza glabra

Linn.)

85

29 Showing macroscopic evaluation of flower of Dhataki

(Woodfordia fruticosa (L.) Kurz.)

88

30 Showing Physiochemical parameter of Dhataki

(Woodfordia fruticosa (L.) Kurz.)

89

31 Showing results of preliminary phytochemical screening of

aqueous extract of Dhataki (Woodfordia fruticosa (L.)

Kurz.)

90

32 Rf values of sample of Dhataki (Woodfordia fruticosa (L.)

Kurz.) Solvent system - Toluene: Chloroform: Ethyl

Acetate: Formic acid (2.0: 6.0: 6.0: 2.0)

91

List of tables


XII

33 Rf values of sample of Dhataki (Woodfordia fruticosa (L.)

Kurz.) Solvent system - Toluene: Ethyl Acetate: Methanol:

Formic acid (3.0: 3.0: 0.8: 0.2)

95

34 Number of patients 98

35 Number of organisms 99

36 Colony characterization and microscopy of Staphylococcus

aureus

100

37 Distribution based on sensitivity of aqueous extracts of

different concentrations of Yashtimadhu against

Staphylococcus aureus

101


different concentrations of Dhataki against Staphylococcus

aureus

103 -104

39 Comparing antibacterial action of aqueous extracts of

Yashtimadhu and Dhataki against Staphylococcus aureus

105

40 Comparing the zone of inhibition of aqueous extracts of

Yashtimadhu and Dhataki against Staphylococcus aureus

106

41 Statistical values of One- Way Anova test for Yashtimadhu

and Dhataki against Staphylococcus aureus

107

42 Statistical values of unpaired t- test for Yashtimadhu and

Dhataki against Staphylococcus aureus

108 -109

43 Colony characterization and microscopy of Escherichia

coli

110



Escherichia coli

111


different concentrations of Dhataki against Escherichia coli

113 -114

List of tables


XIII


Yashtimadhu and Dhataki against Escherichia coli

116


Yashtimadhu and Dhataki against Escherichia coli

116


and Dhataki against Escherichia coli

117


Dhataki against Escherichia coli

119

50 Colony characterization and microscopy of Acinetobacter 120



Acinetobacter

121


different concentrations of Dhataki against Acinetobacter

123


Yashtimadhu and Dhataki against Acinetobacter

125


Yashtimadhu and Dhataki against Acinetobacter

126


and Dhataki against Acinetobacter

127


Dhataki against Acinetobacter

128

57 Colony characterization and microscopy of Pseudomonas

areuginosa

130



Pseudomonas aeruginosa

131

59 Distribution based on sensitivity of aqueous extracts of 133

List of tables


XIV

different concentrations of Dhataki against Pseudomonas

aeruginosa


Yashtimadhu and Dhataki against Pseudomonas aeruginosa

135


Yashtimadhu and Dhataki against Pseudomonas aeruginosa

136


and Dhataki against Pseudomonas aeruginosa

137


Dhataki against Pseudomonas aeruginosa

138

List of figures

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhidravya for Yashtimadhu ( Glycyrrhiza glabra Linn) w.r.t vranahara karma”

XV

FIGURENo

Contents Page No

1.HPTLC photo documentation of ethanol extract ofYashtimadhu 85

2. Densitometric scan of Yashtimadhu

86 - 872a. At 254 nm

2b. At 366 nm

2c. At 620 nm

3.

HPTLC photo documentation of ethanol extract ofDhataki (Toluene: Chloroform: Ethyl Acetate:Formic acid) 91

4.Densitometric scan of Dhataki (Toluene:Chloroform: Ethyl Acetate: Formic acid)

92 - 934a. At 254 nm

4b. At 366 nm

4c. At 620 nm

5.

HPTLC photo documentation of ethanol extract ofDhataki (Toluene: Ethyl Acetate: Methanol: Formicacid) 94

6.Densitometric scan of Dhataki (Toluene: EthylAcetate: Methanol: Formic acid)

95 - 976a. At 254 nm

6b. At 366 nm

6c. At 620 nm

7. Macroscopy of crude drug Yashtimadhu 190

8. Microscopy of Yashtimadhu root

191 - 194

8 a. T.S of root

8 b. A portion enlarged

8 c. Cork, periderm, cortex

8 d. Phloem

List of figures


XVI

8 e. Phloem, cambium, xylem

8 f. Cambium and Xylem

8 g. Xylem and pith

8 h. Cork, periderm and cortex with starch grains

8 i. Stratified cork and cortex

8 j. Xylem

8 k. Phloemfibres

9. Macroscopy of flower of Dhataki 195

10. Microscopy of corolla of Dhataki

196 - 198

10a. T.S of corolla

10b. A portiom enlarged

11. Microscopy of Anther

11a. T.S of anther

11b. Pollen grains

12. TS of flower through ovary

13. TS of Stalk

14. Collection of Dhataki

19915. Coarse powder of Dhataki

16. Coarse powder of Yashtimadhu

17. Soxhlet apparatus – Yashtimadhu

20018. Soxhlet apparatus - Dhataki

19. Water bath

20. Extracts of Dhataki and Yashtimadhu

21. Phytochemical analysis

20122. Loss on drying - Dhataki

23. Loss on drying - Yashtimadhu

24. Water & alcohol extractive value202

25. Ash value

26. Preparation of Media 203

List of figures


XVII

27. Serial Dilution – Concentration

28. Collection of Pus sample

20429. Culturing of Bacteria

30. Sensitivity Test

31. Sensitivity Test 205

32. Zone of inhibition of Dhataki206

33. Zone of inhibition of Yashtimadhu

34. Measurement of Zone of inhibition 207

ABSTRACT


XIX

ABSTRACT

Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhi

dravya for Yashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranahara

karma .

Background- The demand for medicinal plant based raw material is growing at the rate

of 15- 25% annually.1 The degree of threat to natural population of medicinal plants has

increased because more than 90% of medicinal plant raw material for herbal industries in

India is drawn from natural habitat.2 Glycyrrhiza glabra Linn. is one of the most widely

used herb due to its high value index in food grade, pharmaceutical grade , cosmetic

grade, feed grade and other fibres materials.6 The part used is root which is a destructive

form of harvesting.7 The drug is imported from Pakistan, Iran, Afghanistan6 and

adulterated with roots of Abrus precatorius Linn.9 to meet the demand which is upto

5000 tonnes per annum.6 Ayurvedic literature mentions Dhatakipushpa as the substitute

of Yashtimadhumoola.3- 5 Pratinidhi dravya or substitute means substances having

similar pharmacological activities as like that of genuine drug but may not have similar

morphology. Dhataki is found throughout India. Both Yashtimadhu and Dhataki having

pitta shamaka property possesses Vranahara, Vishahara, Trishnahara karma similar to

each other.14- 17 Thus, to prevent adulteration as well to maintain the quality of the drug,

Dhatakipushpa can be used as a substitute for Yashtimadhumoola, thus enriching the

source for Vranahara Karma.

ABSTRACT


XX

Objectives- The present study was aimed to evaluate comparative antibacterial activity

of Glycyrrhiza glabra Linn. and Woodfordia fruticosa (L.) Kurz. from pus of non

healing ulcer patient by culture and sensitivity in vitro.

Material and Methods- The methods was divided into, pharmacognostic and

experimental study. In the pharmacognostic study, an attempt was made by collection

authentication, detail macroscopic, microscopic, physico-chemical, preliminary

phytochemical analysis, HPTLC with its densitometric scan and preparation of extracts of

Glycyrrhiza glabra Linn. and Woodfordia fruticosa (L.) Kurz. was carried out.

In experimental study - invitro , antibacterial activity of aqueous extracts of Yashtimadhu

(Glycyrrhiza glabra Linn.) and Dhataki (Woodfordia fruticosa (L.) Kurz ) was

analyzed against Staphylococcus aureus, Escheriochia coli, Acinetobacter and

Pseudomonas aeruginosa from the pus sample of patient suffering from nonhealing

ulcer.

Result:

Pharmacognostic evaluation of both drugs showed genuinely as per standard. HPTLC

with densitometric scan has showed common active components present in Yashtimadhu

(Glycyrrhiza glabra Linn) and Dhataki (Woodfordia fruticosa (L.) Kurz. ). Both the

drugs have shown significant antibacterial activity in terms of percentage of sensitivity at

various concentration . On comparison, the effect of aqueous extract of Dhataki was

found to be better when compared to aqueous extract of Yashtimadhu. It was also

observed that as the concentration increases the antibacterial activity of both the drug also

increases except for the aqueous extract of Yashtimadhu against Escherichia coli.

ABSTRACT


XXI

Conclusion:

The study has shown that both drugs Yashtimadhu (Glycyrrhiza glabra Linn.) and

Dhataki (Woodfordia fruticosa (L.) Kurz. ) were equally effective in Vranahara karma

(Antibacterial activity).

Keywords – Glycyrrhiza glabra Linn. ; Woodfordia fruticosa (L.) Kurz. ; Antibacterial

activity; Vranahara; Non - healing ulcer; Substitute; Aqueous extract; Pus culture;

Sensitivity.

INTRODUCTION

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhidravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference to vranaharakarma” Page 1

INTRODUCTION

Herbal drug therapy is one of the prerequisites for the success of primary health care in

the community having various culture and tradition like India. Ayurveda is its own

medicine, culture and identifying pillar of country. Plants have always been a common

source of medicament, either in the form of traditional preparations or as pure active

principles. The demand for medicinal plant- based raw materials is growing at the rate of

15 to 25% annually.1 The degree of threat to natural population of medicinal plants has

increased because more than 90% of medicinal plant raw material for herbal industries in

India is drawn from natural habitat.2 It is thus reasonable for decision-makers to identify

locally available plants and their various sources which are used as Pratinidhi dravya or

substitutes.

Pratinidhi dravya or substitute means substances having similar pharmacological

activities as that of genuine drug but may not have similar morphology. Usage of

Pratinidhi dravya or substitute drug becomes inevitable due to unavailability of original

botanical source, adulteration and to preserve the drugs from being endanger.

Ayurvedic literature mentions Dhataki pushpa (Woodfordia fruticosa (L.) Kurz.) as the

substitute of Yashtimadhu moola (Glycyrrhiza glabra Linn.).3-5

Yashtimadhu (Glycyrrhiza glabra Linn.) is one of the most widely used herb due to its

high value index in food grade, pharmaceutical grade, cosmetic grade, feed grade and

other fibres material.6 The part used is root which is a destructive form of harvesting.7

The drug is imported from Asia minor, Iraq, Persia and other Central Asian countries.8

To meet the high demand the drug is over exploited and also adulterated with roots of

INTRODUCTION


Abrus precatorius Linn and roots of Glycyrrhiza uralensis Fish (Manchurian liquorice).

Stem pieces of Glycyrrhiza glabra Linn. are also sold in place of root.9 Glycyrrhiza

glabra Linn. comes under threatened/ depleted (R. N. Chopra’s list).10 Glycyrrhiza

glabra Linn. is an endangered medicinal plant and has been placed in red data book.11

The active constituents of Glycyrrhiza glabra Linn. i.e. Glycyrrhizine cannot be produced

synthetically.12

Dhataki (Woodfordia fruticosa (L.) Kurz.) is found throughout India.13 As both

Yashtimadhu (Glycyrrhiza glabra Linn.) and Dhataki (Woodfordia fruticosa (L.) Kurz.)

having pitta shamaka property possess vranahara, vishahara, trishnahara karmas

similar to each other.14- 17 Dhatakipushpa (Woodfordia fruticosa (L.) Kurz.) is mentioned

in various preparation used in vrana.18- 24 In this study antibacterial activity from pus

sample of patient suffering from Dushtavrana is taken.

DushtaVrana is a common and oftenly encountered problem faced in surgical practice.

The presence of Dushta vrana worsens the condition of the patient with different

complications and may become fatal. Each day new antibiotics are coming to cope up

with the infections. But they are effective up to a certain extent only and become resistant

to themselves. As Yashtimadhu (Glycyrrhiza glabra Linn.) is an effective drug in dushta

vrana and being endangered and is unable to meet the huge demand so its pratinidhi

dravya Dhataki (Woodfordia fruticosa (L.) Kurz.) is screened for its activity. If Dhataki

(Woodfordia fruticosa (L.) Kurz.) is proved to have potent antibacterial avtivity as same

as Yashtimadhu (Glycyrrhiza glabra Linn.) then sufficient preliminary scientific

evidence is generated in invitro study to use Dhataki (Woodfordia fruticosa (L.) Kurz.) as

INTRODUCTION


an abhava pratinidhi dravya for Yashtimadhu (Glycyrrhiza glabra Linn.) on patients of

non healing ulcer.

Hence the present study is taken up with the objectives of Vranahara Karma

(Antibacterial activity). Both Yashtimadhu (Glycyrrhiza glabra Linn.) and Dhataki

(Woodfordia fruticosa (L.) Kurz.)were analyzed by physico-chemical, phyto-chemical

and chromatographic parameters and its antibacterial activity was evaluated through

invitro study.

OBJECTIVES


OBJECTIVES

1. To assess Dhataki (Woodfordia fruticosa (L.) Kurz) pushpa as an effective Abhava

Pratinidhi Dravya for Yashtimadhu (Glycyrrhiza glabra Linn) moola w.s.r to

dushtavrana.

2. To evaluate Dhataki (Woodfordia fruticosa (L.) Kurz) pushpa and Yashtimadhu

(Glycyrrhiza glabra Linn) moola pharmacogonistically.

Literature review


Yashtimadhu

( Glycyrrhiza glabra Linn. )

Yashtimadhu is the well- known drug mentioned in Ayurveda classics, commonly

known as liquorice. It is used in a large number of Ayurvedic formulations like

Dashamoolarishtam, Aswagandharishtam, Khadira gulika, etc . It is having

pharmacological properties such as anti- imflammatory, antiviral, antimicrobial,

antioxidative, anticancer activities, immunomodulatory, hepatoprotective,

cardioprotective effects.11 As per Ayurvedic Pharmacopoeia of India the source plant

of Yashtimadhu is Glycyrrhiza glabra Linn. of Fabaceae family.

Historical background25

Yashtimadhu is known since vedic period itself in different names. Atharva

Parishishta described madhuka and considered it as dourbhagyanasana and

garbhabrmhana. Commentators like Sayana identified madhuka with Yashtimadhu. It

was used in the treatment of animal poison. Madhuyashtika is quoted in the context of

"Mulavidhi”. All Samhitas and Nighantus mentioned about this plant. Brhatrayees

mentioned the use of this herb extensively in therapeutics. Acharya Charaka included

it in many of kashaya vargas and also emphasized its utility among rasayana drugs.

It is one among the four medhya rasayana.

Literature review


Nirukthi / Basonym26

rÉÉ¹ÏÂmÉÇ qÉkÉÑMüÇ CÌiÉ |

Stem of Glycyrrhiza glabra Linn. is sweet.

Synonyms 27-32

It refers to the various names of a drug, they are formed according to the

morphological character, place of origin, habit, action , therapeutic uses, historical

importance etc. It helps for identification of drug and to remember the drug and its

characteristic features.

Table No - 01: Synonyms mentioned by various Samhitha, Nighantu

Sl No. Synonyms DN SON MPN RN KN BPN

1. Madhuka + +

2. Madhuyashti + + + + +

3. Madhuyashtika +

4. Madhuparni +

5. Madhusrava + + +

6. Madhuvalli +

7. Madhu + +

8. Madhoolika +

9. Jalaja +

10. Kleethanakam + +

11. Kleethakam +

12. Klitana +

13. Klitika +

Literature review


14. Rasa +

15. Soshanasini +

16. Soumya +

17. Virasa +

18. Yashti + + + +

19. Yashtimadhu + + + +

20. Yashtimadhuka + + +

21. Yashtika +

22. Yashtyahvam + + + +

Interpretation of Synonyms33-34

Athirasa having more madhura

Kleethakam, Kleethanakam that which cures maleinfertility

Madhuka, Madhuyashtika, Yashyimadhuka,Madhoolika, Madhurasa, Madhuyashti, Madhusrava

the drug is sweet likeHoney

Soshanasini that which cures sosha

Soumya that which is sita invirya

Yashtimadhu, Yashti, Yashtika, yashtiyahva having sweet stem

Literature review


VERNACULAR NAMES35

Names given in different languages. It helps in easy identification of drugs pertaining

to the region.

Table No – 02: Vernacular names of Yashtimadhu (Glycyrrhiza glabra Linn.)

Assami Jesthimadhu, Yeshtmadhu

Bengali Yashtimadhu

English Liquorice root

Gujarati Jethimadha, Jethimard, Jethimadh

Hindi Mulethi, Mulathi, Muleti, Jethimadhu, Jethimadh

Kannada Jestamadu, Madhuka, Jyestamadhu, Atimadhura

Malayalam Irattimadhuram

Marathi Jesthamadh

Oriya Jatimadhu, Jastimadhu

Punjabi Jethimadh, Mulathi

Tamil Athimadhuram

Telugu Atimadhuramu

Urdu Mulethi, Asl – us - sus

Literature review


Classification ( Vargeekarana)36-43

It is the mode of classical differentiation of plants, done by the Acharya on the basis

of morphological characters, pharmacological properties and therapeutic activity of

the plant which helps for easy identification of the plant on the wider basis.

Table No – 03: Classification of Yashtimadhu (Glycyrrhiza glabra Linn.) in

different Ayurvedic classics

Sl

No

Category CS SS AH SON MP

N

RN KN BPN Sha.N

Angamarda

prashamana

+

Asthapanopaga +

Jeevaneeya + +

Kandooghna +

Kantya +

Mutravirajaneeya +

Sandhaneeya +

Snehopaga +

Sonithasthapana +

Vamanopaga +

Varnya +

Anjanadi + +

Kakolyadi +

Saribadi + +

Ambashtadi +

Chardhaneeya +

Haridradi +

Literature review


TAXONOMICAL POSITION44-45

Kingdom -Plantae

Class -Dicotyledonae

Subclass -Polypetalae

Series -Calyciflorae

Order -Leguminales

Family -Fabaceae

Genus -Glycyrrhiza

Species -glabra

Scientific name -Glycyrrhiza glabra Linn.

Niroohana +

Nyagrodhadi +

Gudoochyadi varga + +

Oshadhi varga +

Abhayadi varga +

Pippalyadi varga +

Hareethakyadi varga +

Literature review


Taxonomical Description 25,33,35

Habit: It is a hard, semi- perennial, erect herb or under shrub growing to a height of

upto 1.7m. It has a characteristic pleasant taste.

Roots: Thick, much branched, with yellow or reddish skin and yellowish inside.

Leaves: Imparipinnate and covered with sticky glandular trichomes.

Leaflets: Ovate- lanceolate, occurring in 3-7 pairs, smooth.

Inflorescence: Raceme, shorter than the leaves.

Flowers: 1 cm long, bluish or violet coloured. Calyx are 2 lipped, upper teeth connate

higher than the lower ones. Standard petal is narrow, wings and keel are acute.

Stamens are delphous. Ovary many ovuled.

Fruits: Pods, red to brown in colour, 2.5cm long, compressed.

Seeds: 2-5, deep grey, reniform.

Distribution46

It is distributed in the Sub- tropical and warm temperate regions of the world, chiefly

in the Mediteranean countries, South Europe, Asia Minor, Egypt, Turkistan, Iran,

Siberia, Persia, Arab countries and Afghanistan. In India, it is reported to be

cultivated in Baramulla, Srinagar, Jammu, DehraDun, Delhi and South India.

Literature review


MACROSCOPIC 25, 33,35,45

Outer Surface: Yellowish brown or dark brown, cylindrical, longitudinally wrinkled

pieces, with occasional small buds and encircling scale leaves, transversely, cut and

smoothened surface.

Inner Surface: Shows a cambium ring about one- third of radius from outer surface.

Fracture: Coarsely fibrous in bark and splintery in wood.

Odour: Faint and characteristic.

Taste: Sweetish

MICROSCOPIC 25,33,35,45

Transverse section : Rhizome

Cork: 10-20 or more layers of tabular cells, outer layers with reddish brown

amorphous contents, inner 3 or 4 rows having thicker, colourless walls.

Secondary Cortex: usually of 1-3 layers of radially arranged parenchymatous cells

containing isolated prisms of calcium oxalate.

Phloem: a broad band, parenchymatous cells of inner part cellulosic and outer

lignified; radially arranged groups of about 10 or 50 fibres, surrounded by a sheath of

parenchyma cells, each usually containing a prism of calcium oxalate.

Cambium: form a tissue of 3 or more layers of cells.

Literature review


Secondary xylem: distinctly radiate with medullary rays, 3 or 5 cells wide, vessels

about 80- 200 µ in diameter with thick, yellow, pitted, reticulately thickened walls,

groups of lignified fibres with crystal sheaths similar to those of phloem.

Xylem parenchyma: 2 kinds: those between the vessels having thick pitted walls

without intercellular spaces, the remaining with thin walls.

Pith: parenchymatous cells in longitudinal rows, with intercellular spaces.

Transverse section: Root

Shows structure closely resembling to that of stolon except that no medulla is present.

Xylem: tetrarch, usually four principal medullary rays at right angles to each other.

All Parenchymatous tissues containing abundant, simple, and compound consisting of

2- 4 components, rounded to oval starch grains, measuring 6 to 13 µ in diameter.

POWDERMICROSCOPY 35

Fine powder shows fragments of fibres, vessels with simple and bordered pits; starch

grains simple, oval to rounded, 2- 4 or more components, measuring 6- 13 µ in

diameter.

CHEMICAL CONSTITUENTS47

MAJOR – Glycyrrhizin, glycyrrhizic acid

OTHERS –

Glabranins A & B, isoglabrolide, deoxoglabrolide, glabrolide, glycyrrhetol,

Literature review


21α-hydroxyisoglabrolide; liquoric, liquiritic, glycyrrhetic, 11-deoxo-glycyrrhetic,

21,24-dihydroxy-11-deoxo-glycyrrhetic, 24-hydroxyglycyrrhetic, 18-α-glycyrrhetics

and 18α-hydroxyglycyrrhetic acids; glabranine, pinocembrin, prunetin, glucoliquiritin

apioside, prenyllicoflavone A, shinflavanone, shinpterocarpin, 1-methoxyphaseollin;

licoflavonol, kumatakenin, glycyrol, licoricone; echinatin, glabrol, glisoflavone,

glycyrrhisoflavanone, glyycyrrhisoflavone, semilicoisoflavone B, 1-

methoxyficifolinol, isoangustone A, licoriphenone, kaempferol-3-0-methyl ether;

glyzarin, glyzaglabrin, formononetün, 4'7- dihydroxyflavone, licoisoflavanone,

licoisoflavones A & B, 7-acetoxy-2-methylisoflavone, 7-methoxy-2-methylisoflavone,

7-hydroxy-2-methylisoflavone; licuraside, liqurazid,

liquiritin, licochalcones A & B, isoliquiritin, neoisoliquiritin, 4-hydroxychalcone,

liquiritigenin, isoliquiritigenin, 2',4',4-trihydroxychalcone, 2,4,4'-trihydroxychalcone,

licopyranocoumarin, licoarylcoumarin, licocoumarone; y-nonalactone, cumic alcohol,

indole, anethole, eugenol, estragole, glycyrrhetinic acid.

Table No – 04: RASAPANCHAKA48-54

GUNA DN MPN RN KN BPN NA APIRasa Madhura + + + + + + +

Kinchit Tikta +Guna Guru + + + + +

Snigdha + + + +Virya Sita + + + + + + +Vipaka Madhura + +

Literature review


Table No – 05: KARMA49-52

Karma DN RN KN BPNBalavarnakrit +Chakshusya + +Keshya + +Swarya + +Sukrala +Varnya +Vrishya +

Table No – 06: INDICATIONS 49-52

CULTIVATION TECHNOLOGY 55

It can be propagated by seeds, shoot cuttings, root cutting and by tissue culture.

The traditional method of perpetuation is by utilizing the cuttings prepared from the

old crown of the lifted roots, cut into pieces of 10-15 cm length. The plants are also

Indications DN RN KN BPNChardi + + +Glani +Kshaya + + +Sadyakshatasra +Sopha +Sosha + +Trsna + +Vrna + +Vrnasotha +Visha + +

Literature review


produced from runners or underground stems which are prepared into cuttings 10 cm

long, each with 2 buds. These cuttings are kept in moist sphagnum moss for about 8-

10 days and afterwards, when their buds start sprouting, they are planted in the main

field. The crop occupies the land for a period of 4 to 5 years, and the growth is slow

during the first 2 years.

HARVESTING AND STORAGE 55

The crop is ready for uprooting about 3 to 4 years after planting and just before the

plants have become fruit. The plants arc lifted in autumn (November-December) after

the rains. A trench about 60 cm deep is first dug at the sides of the ridges, then by

working inwards, the soil is loosened from the roots so that they can be pulled out

easily. The aerial parts are cut and removed. The broken root-parts raising the

succeeding crop, only the gaps need to be filled with rooted cuttings.

Roots and underground stems are cut into 15 to 20 cm long pieces, 1-2 cm in diameter

and are dricd alternately in the shade and sun, this may take several months. The

drying process reduces the weight to 50% and the moisture from 50-60%. Artificial

drying can also be done at 30-40°C by using mechanical driers.

STATUS OF THREATENED 56- 57

Glycyrrhiza glabra Linn. comes under threatened/ depleted (R.N. Chopra’s list).

Glycyrrhiza glabra Linn. is an endangered medicinal plant and has been placed in red

data book.

Literature review


ECONOMIC USES 58

It is used throughout country in various recipies and also as a home remedy for cough

and cold. It is also used as a swectening agent in different syrups, since it is one of the

contents. It is used to flavour soy sauce in china and to flavour tobacco products in the

United States. Unani physicians use it as Rabb-i-Sus for chronic cough and sore throat.

Table No – 07: QUANTITATIVE STANDARDS 54

Physic-chemical parameter Standard value

Foreign matter Not more than 2.0 percent

Loss on drying Not more than 12.0 percent

Ash Not more than 10.0 percent

Acid-insoluble ash Not more than 2.5 percent

Alcohol-soluble extractive Not less than 10.0 percent

Water-soluble extractive Not less than 20.0 percent

SUBSTITUTES AND ADULTERANTS 46

Roots of Glycyrrhiza Uralensis Fish. (Manchurian liquorice) and Abrus precatorius

Linn. are often adulterated with liquorice. Stem pieces of Glycyrrhiza glabra Linn.

are also sold in place of root .

Literature review


Abrus precatorius Linn. is very toxic due to an alkaline abrine. The distinguishing

property is that it possesses a disagreeable odour and bitter acrid flavour leaving

faintly sweet after taste. Macroscopically the adulterant is characterized by stone cells.

PHARMACOLOGICAL ACTIVITIES 46

Smooth muscle depressant, antimicrobial, hypolipidaemic, antiantherosclerotic,

antiviral, hypotensive, hepatoprotective, anti- exudative, spasmolytic, antidiuretic,

antiulcer, antimutagenic, antipyretic, antioxidant, anti-inflammatory, anti- nociceptive,

expectorant.

TOXICITY AND SAFETY ASPECTS 59- 60

The intake of higher doses of liquorice (above 50 g/day) over an extended period may

cuse sodium retention, hypertension and cardiac complaints. If taken in excessive

amounts it can cause metabolic disturbances known as pseudoaldosteronism (due to

mineralocorticoid effect of glycyrrhizin) leading to oederna, hypertension and weight

gain.

The drug when used within the recommended dosage and the treatment period is

devoid of any adverse reactions.

ACUTE TOXICITY 59- 60

Glycyrrhizin (crude extract 48-58%):

LD50 values in rats and mice

Literature review


LD50 s.c. 4- 4.4 g/kg

LD50 i.p. 1.42- 1.70 g/kg

LD50 oral 14.2-18.0 g/kg

DOSAGE 54

Powder: 2 to 4 g

RESEARCH PROFILE

1. Antimicrobial potential of Glycyrrhiza glabra Linn. roots byVivek K Gupta, A.

Fatima, ed.al - Genetic Resources and Biotechnology Division , Central Institute of

Medicinal and Aromatic Plants (CSIR) , Analytical Chemistry Division, Lucknow,

2008 :

The activity guided fractionation of ethanolic extract from the roots of

Glycyrrhiza glabra Linn. and subsequent phytochemical

analysisresultedinidentifyingglabridinastheactiveconstituent and hispaglabridin B as

inactive constituent against Mycobacterium tuberculosis.Antitubercular activity of the

glabridin was found to be 20-times higher than the crude extract.The

antimycobacterial activity of root ethanolic extract was observed at 500g/mL against

Mycobacterium tuberculosis H37Ra and H37Rv strains through BACTEC assay. Our

results indicate potential use of licorice as antitubercular agent through systemic

experiments and sophisticated anti-TB assay.

Literature review


2. Role of Yashtimadhu Siddha Taila in the management of Dushtavrana (Bed sore) –

A single case study by S. Das , B. Kalita – Journal of Ayurveda and Integrated

Medical Sciences, 2017 :

A subject age 20 years had an ulcer over sacral region which is about 7×6×1 cm3. The

ulcer has foul smell with swelling along with discharge and the floor was covered

with blackish discolouration of tissue. The treatment given is the wound was irrigated

with normal saline daily and after proper debridement unhealthy dead tissue, the

Yashtimadhu Siddha taila was applied locally and the wound was covered with sterile

gauze. The wound was completely healed after 50 days.

3. Management of Non Healing Ulcer with local application of YashtimadhuGhrita

(Glycyrrhizaglabra Linn.) : A single case study by Jigna R. Patel, Tukaram S.

Dudhamal- Indian Journal of Ancient Medicine and Yoga, 2017:

A male subject of 38 years, presented with non healing ulcer on the lateral aspect of

right ankle joint since two and half year. The local findings revealed a reddish large

ulcer with sloping edges, along with mild serous discharge and blackish discoloration

around lateral malleolus of the right leg. The treatment given is wound was cleaned

with normal saline and then applied YashtimadhuGhrita on ulcer once daily. The

dressing was continued until complete healing achieved and the results was assessed

at regular intervals. Along with local dressing; Manjisthadikwatha and

Rasayanachoorna were administered orally twice a day for 2 months. Wound became

clean with healthy granulation on 21st day, complete wound healing was observed by

2 months.

Literature review


4. Antibacterial activity of Glycyrrhiza glabraLinn. against oral pathogens: an invitro

study by F. Sedighinia, Akbar Sofipour Afshar, et al. AJP , Vol 2, No.3 , 2012:

Objectives: Oral infections and dental caries are still considered as serious public

health problems and inflict a costly burden to health care services around the world

and especially in developing countries.

Materials and Methods: In the present study, we evaluated the antibacterial activity of

Glycyrrhiza glabra Linn. against oral pathogens by diffusion methods and determined

the minimum inhibitory concentration (MIC) by both broth and Agar dilution

methods and minimum bactericidal concentration (MBC) by broth dilution methods.

Results: In this study, G. glabra Linn. extract showed good antibacterial activity

against six bacteria. No strain in this study showed resistance against this extract.

Conclusion: G. glabra Linn. is suggested as an appropriate candidate to help us in

order to control dental caries and endodontic infections.

5. Studies of antibacterial activities of Glycyrrhiza glabra Linn. root extract by Manoj

M Nitalikar, et al. International Journal of Pharm Tech Research, Vol 2, No. 1 , 2010:

The licorice plant (Glycyrrhiza glabra Linn. Family Leguminoceae) has been used by

physician and herbalists since the earliest of times. It is also knows as “sweet roots”,

which contains a compound that is roughly 50 times sweeter than sugar .Many of the

claims for the effectiveness of licorice extracts have been shown by modern science to

be credible, a root component (Glycyrrhizin) being generally regarded as the major

biologically-active principle. Licorice extracts have been widely used in

Literature review


pharmaceutical and confectionery industries because of the presence of glycyrrhizin.

A study was conducted to determine the antibacterial activities of Licorice root

extract in ether, chloroform, acetone on bacteria using the well diffusion method. The

extracts showed significant antibacterial activities against two grampositive (Bacillus

subtili and Stapphylococcus aureus) and two gram-negative (Escherichia coli and

Pseudomonas aeruginosa) bacteria.

6. Phytochemical screening and evaluation of antimicrobial activity of Glycyrrhiza

glabra Linn. by Pankaj Kushwah, et al. PharmaTutor, Vol 2 (5) , 2015

The present study was undertaken to explore the phytochemical screening and

antimicrobial activity of stems of Glycyrrhiza glabra Linn. Glycyrrhiza glabra Linn.

were screened for their antimicrobial activity by agar well diffusion method, standard

screening method and disc diffusion method. In the phytochemical screening showed

presence of secondary metabolites, flavonides, glycosides and terpenoids. It also

showed potent against almost all the test organism. Study indicate that Glycyrrhiza

glabra has antibacterial, antioxidant, antimalarial, antispasmodic, anti inflammatory

and anti-hyperglycemic properties. Various other effects like antiulcer, antiviral,

antihepatotoxic, antifungal and herpes simplex have also been studies. One of the

most commonly reported side effects with licorice supplementation is elevated blood

pressure.

Literature review


Dhataki

(Woodfordia fruticosa (L.) Kurz. )

The drug is described since Vedic literature. The drug is mainly used externally for its

sandhaniya, krimighna, kusthaghna, vranaropana and when administered internally

it acts as balya, stambana, jvaraghna,etc. Few references of using this drug in

Sandhana Kalpana are available in Charaka Samhita and Sushruta Samhita. After

Astanga Sangraha period the drug was being used more in Sandhana Kalpana.

Botanically the the plant is identified as Woodfordia fruticosa (L.) Kurz. belonging to

Lythraceae family.

Historical background

Drug history can be traced under following headings i.e

a. Vedic literatures

b. Samhita literatures

c. Nighantu literatures

Vedic literature – Panini Ashtadhyayi mention about Dhataki as one of the useful herb.

In Amarakosha synonyms of Dhataki are mentioned as Agnijvala, Subhikshe, Dhataki,

Dhatupushpika.61

Samhita literature – A detailed description of Dhataki with regard to its therapeutic

uses, different Kalpanas and indications are available in different Samhitas

Charaka Samhita62: References of using Dhataki flowers in many formulations along

with indications are mentioned. Charaka mentioned about the leaves of Dhataki in the

Literature review


preparation of Kashaya to treat Atisara which is one of the Roopa of Rajayakshma.

Reference ofMadhya preparation which is being prepared out of Dhataki is

mentioned in Madhyavarga. He mentioned specifically Dhataki as a Yoni (precursor)

of Asava (Pushpasava).

Sushruta Samhita63: Sushruta categorised Dhataki under Priyangvadi and

Ambashthadi Gana. Though we can get ample of references about Dhataki pushpa for

Sandhana Karma, he even mentioned therapeutic benefits of flowers in Mamsa vrana,

Gandamala, Bhagandara etc in different formulations.

Asthanga Hridaya64: Herbal medicine stream was fully developed in this period, this

is reflected in the formulation of different Sandhana Kalpana. Along with other

ingredients, the use of Dhataki pushpa in the Sandhana Kalpana is available more in

this period.

Nighantu literature – A detailed description of Dhataki with regards to its

classification, synonyms, therapeutic uses, Rasapanchaka and different indications

have been mentioned in different Nighantu .

Nirukthi / Basonym65 :

SkÉÉÌiÉ iÉÉÌMüiÉÉÌlÉ WûÍxÉiÉÉÌlÉ UqrÉÉÍhÉ mÉÑwmÉÉhÉÏÌiÉ AjÉuÉÉ SkÉÉÌiÉ U¤ÉÌiÉ vÉUÏUqÉÉiÉXMæüËUÌiÉ |

Flowers of Dhataki are very attractive and it is also used as medicines.

Literature review


Synonyms

It refers to the various names of a drug, they are formed according to the

morphological character, place of origin, habit, action , therapeutic uses, historical

importance etc. It helps for identification of drug and to remember the drug and its

characteristic features.

Table No – 08: Synonyms mentioned by various Samhitha, Nighantu66- 72

SSl

No.

Synonyms DN SON MPN RN KN BPN Sha.N

1. Agnijvala + +

2. Bahupushpi +

3. Bahupushpika + +

4. Bhramika +

5. Dhatri +

6. Dhatupushpi +

7. Dhatupushpika +

8. Dhaturanga +

9. Dhavani + +

10. Guchhapushpa +

11. Gucchapushpi +

12. Kumuda + + +

13. Kunjara + + + + + + +

14. Lodrapushpini +

15. Mada + +

16. Madahetu +

17. Madani +

18. Madaneeya +

19. Madhyavasini + + + +

Literature review


Interpretation of Synonyms 73

kÉÉiÉMüÐ – kÉÉiÉÑqÉç MüUÉãÌiÉÌiÉ |

It strengthens the Dhatus or it nourishes the body.

Habitat

mÉÉuÉïiÉÏrÉÉ – mÉÉuÉïirÉmÉëSãvÉã eÉÉiÉÉ |

20. Madhyapushpa +

21. Madhyapushpi +

22. Madhuvasini +

23. Medhavasini +

24. Pamshubhaksha +

25. Parvatiya + + + + +

26. Parvati + +

27. Pramadini +

28. Ratispruha +

29. Rodhrapushpinee +

30. Sanghapushpa +

31. Sanghapushpi +

32. Seedupushpi +

33. Shabdita +

34. Sindupushpi +

35. Subhiksha + + + + +

36. Tamrapushpi + + + + + + +

37. Teevrajvala + +

38. Vahnijvala + + +

39. Vahnipushpa + + +

40. Vahnishikha +

Literature review


Plant commonly grows in hilly region.

xÉÑÍpÉ¤ÉÉ – oÉÉWÒûsrÉãlÉÉãmÉsÉprÉqÉÉlÉÉ |

Dhataki grows abundantly and one plant yield good quantity of flowers.

Morphology

kÉÉiÉÑmÉÑwmÉÏ – kÉÉiÉÑ: aÉæËUMüÇ, iÉSèuÉhÉÉïÌlÉ mÉÑwmÉÉhrÉxrÉÉ:|

Flowers of Dhataki are red in colour like that of Red ochre.

uÉÎlWûmÉÑwmÉÉ – uÉÎlWûeuÉÉsÉãuÉ U£üuÉhÉÉïÌlÉ mÉÑwmÉÉhrÉxrÉÉ:|

The flowers are red in colour like that of flame.

aÉÑcNûÉmÉÑwmÉÉ – aÉÑcNãûmÉÑwmÉÉhrÉxrÉÉ:|

Flowers occur in bunches.

oÉWÒûmÉÑÎwmÉMüÉ – oÉWÕûÌlÉ mÉÑwmÉÉhrÉxrÉÉ:|

Dhataki plant will have many flowers.

Properties and Actions

qÉSWãûiÉÑ: - qÉSãÅÍqÉwÉuÉã WãûiÉÑ: ÌlÉÍqÉ¨ÉqÉç|

Dhataki flowers are used to prepare Alcoholic preparation.

qÉkrÉuÉÉÍxÉlÉÏ – qÉkrÉÇ uÉÉxÉÌrÉiÉÑ vÉÏsÉqÉxrÉÉ: |

Literature review


Dhataki flowers are having property of generating alcohol.

xÉÏkÉÑmÉÑwmÉÏ – xÉÏkÉÉæ qÉkrÉã mÉërÉÑ£üÉÌlÉ mÉÑwmÉÉhrÉxrÉÉ: |

Dhataki flowers are used in Alcoholic formulations.

VERNACULAR NAMES74

Names given in different languages. It helps in easy identification of drugs pertaining

to the region.

Assami Dhaiphool

Bengali Dhai

English Fire flame bush

Gujarati Dhavadi

Hindi Dhai

Kannada Dhataki, Tamrapushpi

Malayalam Tattiripuvu, Tatire

Marathi Dhalas

Oriya Dhatuka Harwari

Punjabi Dhavi

Tamil Dhatari Jargi

Telugu Are puvvu, Sireenji

Urdu Dhaataki Dhatupushpi

Table No – 09: Vernacular names

Literature review


Classification ( Vargeekarana) 75- 85

It is the mode of classical differentiation of plants, done by the Acharya on the basis

of morphological characters, pharmacological properties and therapeutic activity of

the plant which helps for easy identification of the plant on the wider basis.

Table No – 10: Classification of Dhataki (Woodfordia fruticosa (L.) Kurz. ) in

different Ayurvedic classics

Sl

No

Category CS SS AS AH D

N

SON MP

N

RN KN BP

N

Sha.N

Mutra virajaneeya + +

Pureesha sangrahaneeya + +

Sandhaneeya + +

Ambashthadi gana + + +

Priyangvadi gana + + +

Abhayadi varga +

Ashta varga +

Aushadhi varga +

Chandanadi varga + +

Dhatakyadi varga

Haritakyadi varga +

Pippalyadi varga +

Literature review


TAXONOMICAL POSITION 86- 87

Kingdom - Plantae

Division - Magnoliophyta

Class - Magnoliospida

Order - Myrtales

Family - Lythraceae

Genus - Woodfordia

Species - fruticosa

Scientific name -Woodfordia fruticosa (L.)Kurz

Taxonomical Description88- 89

Habit: A strangling leafy shrub reaching 3.6 m ht, branches long and spreading

Bark: Smooth cinnamon-brown, peeling off in fibres, young shoots terete, often

clothed with fine hairs.

Leaves: 5-9 x 1.3-2.5 cm, opposite or subopposite, sometimes in whorls of three,

sessile, ovate-lanceolate, acute, soft velvety above, usually hairy and always nigro-

punctate beneath, base rounded or cordate.

Flowers: Numerous, in short 2-15 flowered cymes from the axils of leaves, pedicels

short, glandular-pubescent. Calyx 1.6 cm long, striate, covered with glandular dots

Literature review


with a small companulate base and a long slightly curved bright red tube which is

slightly contracted above, mouth oblique, teeth about 2.5 mm long, triangular and

acute. Petals slightly longer than the calyx-teeth, narrowly linear, produced at the

apex to a long fine point.

Fruits: Capsule 1cm long, usually splitting, the calyx near the base, irregularly

dehiscent.

Seeds: Cuneate-obovoid, brown and smooth.

Distribution90

Found throughout India, ascending to 1700m, particularly common in deciduous

forests and scrub jungles

MACROSCOPIC91

Flower is about 1.2 cm long, occurs as singles or in bunches of 2-15:, calyx 1.0- 1.6

cm long, ridged and glabrous, bright red when fresh but fades on drying, with

campanulate base and oblique apex having 6 triangular and acute teeth; each tooth

being, 2-2.25 mm long; 6, very minute accessory sepals attached outside at the

juncture of calyx tooth and deeper in colour; petals 6, attached inside the mouth of

calyx-tube, slightly longer than calyx tooth, alternating with calyx-tooth, pale rose or

whitish, thin, papery, lanceolate, acuminate; stamens 12, united at the base, about 1.5-

2 cm long, filament filiform, curved at the apex, keeping anthers inside calyx-tube;

Literature review


anthers dorsifixed, brown, almost rounded or broadly ovate; carpels 2, united,

ovary superior, style filiform, longer than ovary and stamens; taste astringent.

MICROSCOPIC 91

Transverse section of sepal shows single layered cuticularised epidermis, provided

with both glandular and covering trichomes; glandular trichomes multicellular, long,

consisting of a stalk and a globose, thin walled, multicellular head; covering

trichomes unicellular, thick walled, broad at the base and pointed at the apex. Ground

tissue consists of thin walled, parenchymatous`cells. Surface view of petal shows thin

walled, parenchymatous cells provided with very few sparsely distributed covering

trichomes. Transverse section of filament shows epidermis consisting of single

layered, tangentially elongated cells, covered with a very thick cuticle. Ground tissue

consists of thin walled parenchymatous cells with intercellular spaces, surrounding a

central vascular cylinder of spirally thickened vessels. Transverse section of anther

shows single layered epidermis, covered with cuticle followed by several layers of

thickened cells, surrounding both the pollen sacs having numerous pollen grains;

pollen grains roughly tetrahedral with three pores; central region consisting of thin

walled cells embedding vascular bundle.

POWDERMICROSCOPY 91

Powdered drug shows thick walled pitted epidermal cells provided with papillae;

covering and glandular trichomes, striated cuticle; parenchymatous cells thin walled,

filled with pigments and spheroidal crystals of calcium oxalate; round, oval or

Literature review


polygonal cells with secondary wall thickenings forming the endothecium of anther;

vessels long with spiral and scalariform thickenings; fibres- thick walled, pitted and

with straight or wavy longitudinal wall; pollen grains prolate, tricolporate and starch

grains.

CHEMICAL CONSTITUENTS 92

Major

Woodfordin A, B, C1,2.

Others

Oenothein A3 and B1,2, woodfordin D3, E, F, G, H, I4, isoschimawalin A4, ellagic acid,

myricetin-3-galactoside, pelargonidin-3,5-diglucoside5, cyanidin 3,5-diglucoside6,

quercetin-3-rhamnoside, naringenin-7-glucoside, kaempferol-3-glucoside, hecogenin,

mesoinositol7, norbergenins8, β-sitosterol and chrysophanol-8-0-β-D-

glucopyranoside9.

RASAPANCHAKA79- 85

Sl.

No

GUNA DN MPN RN KN BPN SHN

1. Rasa Katu + + + + + +

Kashaya + + +

2. Guna Laghu + + + +

Manda +

Ushna + + +

Table No - 11: Rasapanchaka

Literature review


KARMA79- 83

Table No – 12: Karma

Karma DN MPN RN KN BPN

Atisarahara + + + + +

Deepana + +

Garbhasthapani +

Krimighna + + +

Madakarari +

Madakrut + +

Pathya + +

Pittala + +

Raktadoshahara

Vishanashani + + + + +

Vrananashani +

Mrudu

3. Veerya Sheeta + + + +

Ushna + +

4. Vipaka katu + + + + + +

Literature review


INDICATIONS 79. 81, 83

Table No – 13: Indications

Indications MPN KN BPN

Atisara + + +

Jantu +

Krimi + +

Raktapitta + + +

Trushna + + +

Visarpa + + +

Visha + + +

PROPAGATION AND CULTIVATION 93

The plant can grow on variety of soils and climatic conditions, but prefers open dry

areas and rocky as well as clayey soil. It propagates mainly by branch cuttings and

seeds. The effect of growth hormones for vegetative propagation of W. fruticosa was

studied. Out of three growth hormones (IAA, IBA and NAA) tried, application of 200

ppm concentration of IBA for 24 hours was found to promote rooting in branch

cuttings. Because of the rock- bearing nature, the plant is also selected for cultivation

for afforestation and reclaiming the barren hill rocks. Germination of seed in sand was

found to be better than in brick powder and soil. The seeds stored at normal room

condition showed a decline in viability from initially 96 % to 1.25 % in 12 months. A

Literature review


rapid propagation method comprising initiation of in vitro shoot tip culture from field

grown flowering plants and reculture of the nodal segments of regenerated shoots in

SH medium was developed for W. fruticosa. Highest multiplication (26-35 shoots)

was recorded when using culture initiation media with 0.5 mg/1 each of BAP and

NAA followed by subculture in 0.2 mg/l BAP. Shoot multiplication rate was further

accelerated by reculturing 0.4-0.6 cm nodal segments of regenerated shoots in media

with 1.0 mg/1 BAP. Regenerated plants, displayed uniform morphological growth and

flowering characteristics.

QUANTITATIVE STANDARDS 94

Table No - 14: Quantitative standards

Physic-chemical parameter Standard value

Foreign matter Not more than 2.0 percent

Loss on drying 25.650 percent (database V-3)

Ash Not more than 10.0 percent

Acid-insoluble ash Not more than 0.5 percent

Alcohol-soluble extractive Not less than 8.0 percent

Water-soluble extractive Not less than 20.0 percent

SUBSTITUTES AND ADULTERANTS

_

Literature review


PHARMACOLOGICAL ACTIVITIES 95

Antipyretic, antibiotic, abortifacient, antifungal, antitumour, antiviral.

SAFETY ASPECTS 94

The drug used traditionally in prescribed doses may be considered safe.

DOSAGE 96

Powder of the flowers: 3 – 6 g

RESEARCH PROFILE

1.The antimicrobial activity of essential oil and plant extracts of Woodfordiafruticosa

by RajandeepKaur,HarpeetKaur – C T Institute of Pharmaceutical Sciences, Jalandhar

(Punjab), 2010 :

Two gram positive bacteria Staphylococcus aureus and Bacillus subtilis; and two

gram negative bacteria Escherichia coli and Pseudomonas aerogenosa were used in

the study. The essential oil obtained from the leaves possessed activity against two

bacteria (P. aerogenosa, B. subtilis) . The antimicrobial activity of plant extracts was

determined by agar disc diffusion method. The microorganisms that were used for the

tests were sensitive to the all three plant extracts (Hexane, Acetone, Methanol). It was

found that the hexane extract of W. fruticosa showed maximum activity against P.

aurogenosa . However the hexane and acetone extract of W. fruticosa showed

minimum activity against B.subtilis .

Literature review


2. In vitro antibacterial activity of the crude methanol extract of

WoodfordiafruticosaKurz. flower (Lythraceae) by J. Parekh, S. Chanda - Brazilian

Journal of Microbiology , 2007:

In vitro antibacterial activity of the crude methanol extract was studied against 15

bacterial strains (six Gram - positive and nine Gram – negative bacteria) by the agar

well diffusion method. The methanol extract of the flower exhibited antibacterial

activity at the varied levels except against Bacillus subtills and Macrococcusflavus.

The methanol extract of the flower was more active against Pseudomonas

pseudoalcaligenes. The extract was more active against Gram- negative bacteria as

compared to Gram- positive. The inhibitory effect of the extract was compared with

standard antibiotics, amoxicillin and ciprofloxacin. The results from this study reveals

that the crude methanol extract of Woodfordiafruticosa contain certain constituents

like tannins with significant antibacterial property which enables the extract to

overcome the barrier in Gram – negative cell wall.

3. Antimicrobial activity of useful parts of Woodfordiafruticosa (Linn.) Kurz.of

Nepal by S. Bhattarai - International Journal of Pharmaceutical & Biological 2011 :

Six extracts (two each of methanol, chloroform and hexane) prepared from leaf and

flower samples of Woodfordiafruticosa Linn were screened for antimicrobial activity

against 14 microorganisms by disk diffusion method.Among six extracts examined,

66% extracts showed antimicrobial property against Bacillus subtilis; 50% extracts

against Staphylococcus aureus, Salmonella Typhi, Salmonella paratyphi,

Citrobacterfrendii each; 33% extracts against Pseudomonas aeruginosa, Escherichia

Literature review


coli, Proteus mirabilis, Klebsiellapneumoniae, Shigelladysenteriae each, and 16%

extracts against Enterobacter spp., Acenitobacter spp., each.Rest of the two human

pathogenic fungi, Candida albicans and Aspergillus spp., did not show any zone of

inhibition against any extracts tested.Extracts were more likely to inhibit Gram-

positive bacteria with respect to Gram-negative bacteria.

4. Antibacterial Potential of Extracts of Woodferdia fruticosa Kurz on Human

Pathogens by M.V. Kumaraswamy, H.U. Kavitha and S. Satish, World Journal of

Medical Sciences 3 (2), 2008:

Woodferdia fruticosa Kurz was tested for antibacterial activity against fourteen

human pathogenic

bacteria. The dried flowers were extracted with deferent solvents viz., petroleum ether,

chloroform, methanol, ethanol and water using soxhlet apparatus. All the solvent

extracts were evaporated to dryness using rotary flash evaporator. Dry residue was

dissolved in respective solvents (1:10 w/v) and tested for antibacterial activity. The

result revealed that among five solvents tested, petroleum ether extracts showed

significant antibacterial activity when compared with Gentamicin for human

pathogens.

5. Wound healing potential of flower extracts of Woodfordia fruticosa Kurz by Neeraj

Verma, G Amresh, P K Sahu, et al. Indian Journal of Biochemistry and Biophysics,

Vol 50, 2013:

Literature review


Wound healing or repair is the body’s natural process of regenerating dermal and

epidermal tissue. Inthis study, we evaluated antimicrobial activity of petroleum ether,

chloroform, ethanolic and aqueous extract against a diverse range of gram +ve and

gram –ve bacteria along with pathogenic fungi. It was evaluated at dose levels of 250

and 500nmg/kg body wt in rats by excision, incision and dead space wound healing

modelsalong with histopathology of wound area of skin. The ethanolic extract extract

showed potent wound healing activity, as evident from the increase in the wound

contraction and breaking strength in dose- dependent manner. The ethanolic extract

exhibited a strong and broad spectrum antimicrobial activity as compared to other

extracts. It showed very low MIC values and inhibited the growth of E. coli,

Staphylococcus aureus and Candida albicans in concentration of 2.5 µg/ disc.

6. Phytochemical and Chromatographic studies in the flowers of Woodfordia

fruticosa (L) Kurz by A. Finose and K.Devaki. Asian Journal of Plant Science and

Research, Vol1 (3), 2011:

The present study primarily aims to carry out a preliminary phytochemical screening

so as to detect the major class of compounds present. TLC profiling of the

Woodfordia fruticosa flowers was carried out using sequential extracts of solvents

with varying polaity; petroleum ether, chloroform and methanol respectively. The

TLC documentation was done in short UV(254nm), long UV(365nm) and visible

light after derivatisation with Anisaldehyde Sulphuric acid as the spray reagent.. Then

DPPH free radical scavenging assay was carried out in the flowers so as to detect its

antioxidant activity. The flower has a significant sweetness when tasted; so the total

Literature review


estimation for the starch content in the plant was carried out. The HPLC studies were

performed in the methanolic extract of the plant, since it gave better separation than

the other two solvents. The results obtained can be used for the genuine identification

of the plant from its adulterants.

7. Pharmacognostic characterisation of flowers Woodfordia fruiticosa Kurz. (Dhataki

Pushpa) used as fermentation initiators by Mallikarjun Admani, KN Sunil Kumar,

Suma V Mallya. Journal of Ayurvedic and Herbal Medicine, Vol 1(1), 2015

Background: Woodfordia fruiticosa Kurz. flowers are highly valued medicinal

material used in Indian System of Medicine. They are used as fermentation initiators

in medicated alcoholic preparations like asavas and arishtas. In addition to this,

flowers are recommended in acute diarrhoea, haemorrhages, ulceration and erysipelas.

Authentication of herbal drug by macro-microscopic and chemical characterization

should be the primary criteria prior to its usage.

Materials and Methods: In the current study flowers of W. fruiticosa are collected and

subjected for macro-microscopic and physico-chemical analysis aiding standard

methodology.

Results: Macro-microscopic features of different parts of a flower are documented

along with their photographs. Physico-chemical values like total ash, acid insoluble

ash, water soluble ash, ethanol soluble extractive and water soluble extractive are

recorded.

Conclusion: Macro-microscopic atlas; along with physico-chemical value serve as

reference standard for identification and distinguishing the sample from its substitutes

and adulterants.

Literature review


VRANA

Definition

The word Vrana derived from Vrana Gatra Vichurnana.97 Gatra means tissue (body

part) Vichurnana means destruction, disruption of continuity break, rupture and

discontinuity. Discontinuity destruction/break/rupture of the body tissue or part of the

body is called Vrana. There is discoloration at the site of Vrana after healing.98 Sa

Vranoti Acchadayati Yasmat Syaat Vrana Iti99 means it covers or conceals the under

lying tissues. As the scar of a wound never disappears even after complete healing

and lives imprint lifelong is called Vrana.100

Nidana of vrana101

The causes or Nidana of Vrana are same as the factors responsible for the vitiation

of Doshas. These are classified as Aaharaja and Vihaaraja. They are as follows

Table No- 15: Nidana of Vrana

Dosha Ahara Vihara

Vata Laghu, Katu, Kashaya, Tikta, Ruksha ahara,

Shaaka, Vallura etc

Bala Vigraha, Ati Vyayama,

Ratri Jagaran, Langhana

Pitta Katu, Amla, Lavana, Tikshna, Ushna,

Laghu, Vidaahi, Tila Taila, Pinyaka,

Kulatha etc

Krodha, Shoka, Bhaya,

Aayasa, Upavasa, Maithuna

etc

Kapha Madhura, Amla, Lavana, Snigdha, Divaswapna, Avyayama,

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Agantujanidana102

Parusha, Pashu, Pakshi, Vyala, Sareesrapa, Prapatana, Peedana, Prahara,

Agni, Kshara, Visha, Teekshna ushadha prayoga , Shakala, Kapala prahara

abhigata, Shringa, Parashu, Shakti, Kunta, Abhighata.

Classification

I. Based on Avastha, Vrana can be classified into

1. Dushta vrana

2. Shuddha vrana

II.Based on Stage of Healing, Vrana can be classified into

1. Ruhyamana Vrana

2. Samyak Roodha Vrana

Shuddha Vrana 103

The Shuddha is devoid of all the three Doshas. Usually the Shuddha Vrana

does not need any treatment whereas Dushta Vrana is difficult to treat. The

floor of the Vrana should be at surface level. The discharge and pain should

be absent. The explanation and Lakshanas of Shudhha Vrana according to

Picchila,Masha, Godhuma etc Aalasya

Rakta Drava, Snigdha, Guru Krodha, Anala and Atapa

Sevana, Shrama etc

Literature review


Sushruta and Vagbhata104 are almost similar. Charaka105 and Madhava

Nidana 106 also explained the features of Dushta Vrana.

Ruhyamaana Vrana Lakshana 107

This is the healing stage of Vrana. Vrana which has Kapotha Varna

and has Sthira Pitika is said to be Ruhyamaan Vrana. Similar type of

description is mentioned by Vagbhata and in Madhava Nidana.

Samyak Roodha Vrana108

Vrana which has healed in its seat (dwelling place) without eruptions

(Granthi) pain (Vedana) or swelling and has the colour as that of Twak

and is even is said to be Samyak Roodha Vrana. Charaka also

mentioned classification of Vrana.109

DUSHTA VRANA

Dushta means getting vitiated by Doshas 110

Dushta means there is localization of Doshas or getting vitiated by Doshas. Vrana

which smells badly (foul odour) has abnormal color with profuse discharge, intense

pain and takes long period to heal is said to be Dushta and the features opposite to

that are of Shuddha Vrana. In this context we can understand it as a nonhealing ulcer.

The features of Dushta Vrana will vary according to the predominant Dosha present

in it.

Literature review


Lakshanas of Dushta Vrana

Lakshanas depending upon the shape, discharge, consistency and

chronicity according to various Acharyas.

Table No- 16: Lakshanas of Dushta Vrana

Sushruta 111 Charaka 112 Ashtanga

Hridaya113

Madhava

Nidana 114

Sarangadhara

Samhita 115

Extremaly narrow or

wide mouthed.

Too soft.

Elevated or depressed.

Black, red or white

coloured.

Too cold or hot.

Full of Pooti

Pooya,Sira,Snayu,Pooti

Pooya Sraavi.

Upward or oblique

course of suppuration. Pus

runs in to cavity and

fissures, having foul

No specific

Lakshanas

mentioned by

Charaka, But

by classification

it is

characterized in

to 12

White

Depressed path

Too thick

Too yellow,

blue, blackish

grey.

Too hard

or soft.

Too

elevated

or too

inverted.

Too cold

or too hot.

Color of

Vrana is

red / black

Severe

painful.

Burning

Purulent

profuse

blood

stained

discharge.

Large

cavity.

Foul

smelling.

Severe

pain.

Opposite

lakshanas

Opposite

lakshana of

Shuddha

Vrana

Literature review


Pancha lakshanas

The Pancha lakshanas are Varna, Vedana, Srava, Gandha, Akruti

Vrana Varna 116

Table No- 17: Vrana Varna

Dosha Vrana Varna

Vata Bhasma, Kapotha, Asthi, Aruna and Krishna

Varna

Pitta Neela, Peeta, Haritha, Syava, Krushna, Rakta,

Kapila, Pingala

Kapha Sweta, Snigdha, Pandu

Rakta Same as Pitta

Sannipataja Sannipataja

smell.

Burning sensation, redness

and itching. Pustules crop

up around, secrete blood

Black foul

smelling. Wide

cavity filled

with pus.

Narrow mouth

sensation.

Inflamed.

Redness

and itching

is present.

Chronic in

nature

of Shuddha

Vrana

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Vrana Gandha

Table No- 18: Vrana Gandha according to Sushruta 117

Dosha Vrana Gandha

Vata Katu Gandha

Pitta Teekshna Gandha

Kapha Ama Gandha

Rakta Loha Gandha

Sannipatha Katu, Teekshna and

Ama Gandha

Vata Pitta Laja Gandha

Vata Kapha Athasi

Pitta Kapha Taila

Table No- 19: Vrana Gandha according to Charaka 118

Ghrita Pootika

Taila Amla

Vasa Shyava

Pooya Rakta

Literature review


Vrana Srava 119

Table No- 20: Vrana Srava according to Sushruta

Table No- 21: Vrana Srava according to Charaka 120

Vrana sthana Vata Pitta Kapha Sannipata

Twak Parusa Gomedaka Navanita Nalikerodaka

Mamsa Syava Gomuthra Kasisa Ervaruka rasa

Sira Avasyaya Bhasma Majja Kanjika

Snayu Dadhimastu Sankha Pishti Arukodaka

Asthi Ksharodaka Kasaya Tila Priyangu phala

Sandhi Mamsadhavana Madvika Nalikerodaka Yakrut

Koshta Pulakodaka Tailam Varaha Vasa Mudgayusha

Lasika Jala

Pooya Asruk

Aruna Haridra

Kashaya Pinjara

Haritha Neela

Rooksha Snigdha

Sita Asita

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Vrana Vedana121

Table No- 22: Vrana Vedana

Vrana Akruti122

Sushruta mentioned 4 normal shapes for ulcer. Others are having irregular

shapes and they are difficult to treat. The Vrana Akrutis are Ayata,

Chaturasra, Vrutta, and Triputaka.

Vata Toda, Bheda, Tadana, Chedana, Ayama, Mantana, Vikshepana,

Chimachimayana, Nirdahana, Avabhajana, Spotana, Vidarana,

Utpadana, Kampana, Purana, Sthambana, Akunchana and various other

types of pain

Pitta Osha, Chosa, Paridaha, Dhoomayana and pain similar to alkaline

substances put on Vrana

Kapha Kandu, Gurutwa, Suptata, Alpa Vedana

Rakta Same like Pitta

Sannipata Lakshanas of all three Doshas

Literature review


Upadrava (Complications)

They may be pertaining to the ulcer or pertaining to the patient. Vranasya

Upadrava123(pertaining to the ulcer) are Gandha, Srava, Varna, Vedana,

Akruti.Vranithasya Upadrava (pertaining to patient) are some systemic

diseases that occur as complication of Vrana which are mentioned by

Sushruta124 and Charaka125

Sadhya – Asadhyata

Characters of Sukha Sadhya Vrana126,Kricchra Sadhya Vrana127,Yapya

Vrana128 Asadhya Vrana129 was explained by Sushruta Samhita.

Vrana Chikitsa

Vrana Chikitsa should be done in Vranitagara130. Sapta Vidha Upakrama131

Shasti Upakrama132 was explained by Sushruta. Treatment principles of

dushta vrana also explained by Sushruta133,Charaka134, Kashyapa135,

Ashtanga Sangraha andAshtang Hridaya136.

Pathya - Apathya137

Sushruta has mentioned Pathya and Apathya Ahara and Vihara for Vranita.

Literature review


Definition of wound:

“The term Wound is break in the continuity of soft parts of body structures

caused by violence of trauma to tissues”.

- Taber‟s Medical Cyclopedia

Definition of ulcer:

“Ulcer word is derived from the Latin word “ulcus”. It means an open sore or

lesion of the skin or mucous membrane accompanied by sloughing of

inflamed necrosis tissue”.

- Taber‟s Medical Cyclopedia

The word wound and ulcer are used synonymous though there are some

similar & dissimilar features.

Literature review


ULCERS

An ulcer is a discontinuity of an epithelial surface (skin or mucous

membrane).It may follow molecular death of surface epithelium or it‟s

traumatic removal, there is usually progressive destruction of surface tissue

cell by cell, as distinct from death of macroscopic portions (e.g.

gangrene/necrosis).138

Chronic ulcers are the wounds that fail to heal, in general they have a fibrotic

margin and a bed of granulation tissue which may include areas of slough

(necrotic tissue)

PARTS OF AN ULCER

1. Margin: It is the junction between normal epithelium and ulcer. It

may be regular or irregular, rounded or oval.

2. Edge: Edge is the one which connects the floor of

ulcer to the margin. Different edges are as follows

Sloping edge: It is seen in a healing ulcer. Its inner part is red because

of healthy granulation tissue. Its outer part is white due to scar or

fibrous tissue. Its middle part is blue due to epithelial proliferation.

Undermined edge: Is seen in spreading type of ulcer like tuberculous

ulcer, were disease process advances in the deeper plane.

Punched out edge: It is seen in gummatous ulcer, and in trophic ulcer.

Raised and pearly white beaded edge: It is seen in a rodent ulcer.

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Everted edge (rolled out edge): It is seen in a carcinomatous

ulcer due to proliferating malignant tissues over the normal skin.

3. Floor: It is the one which is seen. Floor may contain discharge, granulation

tissue.

4. Base: Base is the one on which ulcer rests. It may be bone or soft tissue.

CLASSIFICATION OF ULCERS

Two types of classification of ulcers are possible

1) Clinical 2) Pathological

A.Clinical Classification139Table No- 23: Types of ulcer

Spreading ulcer Healing ulcer Callous ulcer

Surrounding Skin of ulcer

is inflamed, floor covered

with slough without any

granulation tissue.

Floor granulation is

present, Edge bluish

outline of growing

epithelium and slight

serous discharge.

Floor-pale granulation

tissue in durations present

at base, edge, surrounding

skin. Ulcer shows no

tendency towards healing.

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B.

Pathological Classification 140

Non-specific ulcers Specific ulcers

Malignant ulcers

So according to the cause these ulcers are classified as below-

1. Non-specific ulcers: Traumatic ulcer, Arterial ulcer, Venous ulcer,

Neurogenic ulcer, Infective ulcer, Tropical ulcer, Cryopathic ulcer, Martorell‟s

ulcer, Bazin‟s ulcer, Diabetic ulcer, Miscellaneous ulcers.

2. Specific ulcers: These are seen in T.B, Syphilis, Soft sore, Actinomycosis.

3. Malignant ulcers: Epithelioma, Rodent ulcer, Malignant melanoma.

STAGES OF ULCER

The life history of ulcer consists of 3 phases141

1) Stage of extension 2) Stage of transition 3) Stage of repair

1. Stage of extension: During this stage floor is covered with exudates and

slough, while base is indurated. The discharge is purulent and even blood

stained.

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2. Stage of transition: This prepares for healing. Floor becomes cleaner,

slough separates, induration of base diminishes and discharge becomes more

serous. Small reddish areas of granulation tissue appear on the floor.

3. Stage of repair: It consists of transformation of granulation to fibrous tissue

which gradually contracts to form a scar. The epithelium gradually extends

from the new shelving edge to cover the floor. This healing edge consists of

three zones. Outer layer of epithelium appears white. Middle one appears

bluish in colour. Inner reddish zone of granulation tissue covered by single

layer of epithelial cells.

Literature review


ANTIBACTERIAL ACTIVITY

Microbiology is the study of microscopic organisms such as bacteria, viruses, fungi

and protozoa. These microscopic forms of life are present in abundance in the

environment. They are found in water, food, soil and air. Some of the microorganisms

are harmful and some others benefit by association with the biological activity of the

host. An antibacterial agent are a group of materials that fight against pathogenic

bacteria. Thus, by killing or reducing the metabolic activity of bacteria, their

pathogenic effect in the biological environments will be minimized. Antimicrobial

susceptibility testing can be used for drug discovery, epidemiology and prediction of

therapeutic outcome. Most of the antibiotics are in danger of losing their efficacy

because of the increase in microbial resistance. Currently, its impact is considerable

with treatment failures associated with multi-drug resistant bacteria and it has become

a global concern to public health. For this reason, the discovery of new antibiotics is

an exclusively important objective. Microbial and plant products occupy

a major part of antimicrobial compounds discovered until now.

Antibacterial Mode of action142

Different antimicrobial has a different mode of action, owing to the nature of then

structure and degree of affinity to certain target sites within the bacterial cells. Five

basic mechanisms of antibiotic action against bacterial cells include

• Inhibition of cell wall synthesis

•Inhibition of protein synthesis

• Alteration of cell membranes

Literature review


• Inhibition of Nucleic acid synthesis

• Antimetabolite activity.

Antimicrobial agents kill bacteria by different methods depending on the type of

bacteria. Most of the antibacterials kill bacteria immediately by causing the bacterial

cell to explode and is termed as bacterial conjugation. This prevents the multiplication

of the bacteria.

Bacteria

Staphyłococcus aureus143

They are spherical cocci about 0.8 to 1.0 µm in diameter. They are arranged

characteristically in grape-like clusters. They are non-motile, non-sporing, non-

capsulated and Gram-positive. They may also be found singly, in pairs and in short

chains or three or four cells. They are aerobes and facultative aerobes. They grow

readily on ordinary media within a temperature range of 10 - 42°C, the optimum

being 37°C and pH 7.4 to 7.6. On nutrient Agar (24 hours incubation) the colonies are

large circular, convex, smooth, shiny, opaque and easily emulsifiable. Most strains

produce golden-yellow pigment, some strain may form white colonies. The confluent

growth presents a characteristic oil-paint appearance on the nutrient Agar slope.

Most strains are haemolytic, produce a beta type of haemolysis. They grow on Mac

Conkey's medium, producing smaller colonics that are pink due to lactose

fermentation. They are catalase positive and usually hydrolyse urea, reduce nitrates to

nitrites, liquefy gelatin. Staphylococci are killed by a temperature of 60°C in 30

minutes. They resist 1% phenol for 15 minutes. Mercury per chloride 1% solution

Literature review


kills them in 10 minutes. Staphylococcus aureus possesses a large number of cell

wall-associated and extracellular toxins and enzymes which contribute to the ability

of the microorganism to overcome the body's defence and to survive and produce

diseases in the host. Staphylococcus aureus produces cutaneous and deep infections,

exfoliative diseases, food poisoning and toxic shock syndrome.

Escherichia coli 144

E. coli is a Gram-negative, non-capsulated bacillus measuring 1 - 3 µm x 0.4 - 0.7 µm

in size. It is motile by peritrichate flagella but some strains may be non-motile. Spores

are not formed. Capsules and fimbriae are found in some strains. It is an aerobe and a

facultative anaerobe growing on simple media. The optimum temperature for its

growth is 37°C (10-45°C). Colonies are large, thick, greyish white, smooth, opaque or

partially translucent on ordinary media. Colonies on Mac Conkey Agar medium are

red or pink in colour. It ferments lactose, glucose, sucrose, maltose and mannitol

forming acid and gas. It shows negative results in for H2S production, urease test,

gelatin liquefaction, and growth in the presence of KCN. E. coli is Indole and MR

positive and VP and citrate negative. E. Coli possesses 4 types of antigens - Flagellar

(H) antigen, Somatic (O) antigen, Capsular (k) antigen and fimbrial (F) antigens.

Some strains of E. coli produce enterotoxins, haemolysin and verocytotoxin.

E. coli forms a part of the normal intestinal flora of man and animals. Different types

of clinical syndromes are caused by E. coli such as urinary tract infection, diarrhoea,

pyogenic infections and septicemia.

Literature review


Pseudomonas aeruginosa145

It is a slender Gram-negative bacillus, 1.5 - 3.0 µm x 0.5 µm arranged singly, in

pairs or short chains. It is actively motile by połar flagellum. It is aerobic

growing on simple nutrient media with an optimum temperature of 37 C. On

nutrient Agar, the colonics are large 2 - 3 mm in diameter, smooth, translucent,

irregularly round and emit a characteristic fruity odour. It grows on Mac Conkey

and DCA media forming non-lactose fermenting colonies. Many strains are

haemolytic on blood agar. It forms dense turbidity with a surface pellicle in

broth medium. Pseudomonas aeruginosa produces pyocyanin, pyorubrin and

pyomelanin pigments. It is catalase, Arginine dihydrolase, gelatinase and

oxidase positive and indole MR, VP and H2S are negative. Pseudomonas

aeruginosa survives well in a wet environment but is not very resistant to drying.

It is easily killed by heating at 55°C for one hour and it shows high resistance to

chemical disinfectants. Commonest infections caused by Pseudomonas

aeruginosa are urinary tract infections, acute purulent meningitis, respiratory

infections, septicaemia and endocarditis. It may cause wound and burn infection,

eye infection, chronic otitis media and acute necrotising vasculitis which leads

to haemorrhage, infarction of skin and internal organs.

Acinetobacter146

Acinetobacter species are gramnegative bacteria and saprophytes, found in soil,

water and sewage and occasionally as commensals of moist areas of human skin and

Literature review


mucous membrane (oropharynx). These are increasing in importance as opportunist

pathogens. Hospital- acquired infections, including pneumonia, other respiratory

infections; infection of wounds and urinary tract, are most commonly associated with

A. baumanni.

Acinetobacters are nonmotile, usually coccobacillary or coccal in appearance and

diplococcal forms . They grow well on common media and do not reduce nitrates,

some species are oxidase positive.

Culture and sensitivity:

Culture Media:147 A nutrient medium which is a solid or liquid preparation used to

grow, transport, and store microorganisms in a laboratory. Specialized media are

essential in the isolation and identification of different microorganisms, testing of

antibiotic sensitivities and other activities.148 Microbes that are introduced into a

culture medium to initiate growth are called an inoculum. The microbes that grow and

multiply in or on a culture medium are referred to as a culture.

Streak culture method:149 It is the isolation method used to get pure cultures. With

an inoculating loop or swab, inoculums is transferred to the edge of an agar plate

and then streaked out over the surface in one of several patterns.150

Microbial Growth on Solid Media:151 Individual species may form colonies of

characteristic size and appearance often. So inspecting characteristics over microbial

growth on media will aid in identifying microorganisms. Patterns and colony size

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depend on many factors including nutrient diffusion and availability, bacterial

chemotaxis, presence of liquid on surface and hardness of agar.

The Growth of Bacterial Cultures152: The time required for a cell to divide is

called the generation time. Most bacteria have a generation time of 1 to 3 hours;

others more than 24 hours. It varies considerably among organisms and with

environmental conditions, such as temperature. There are four basic phases of

growth pertaining to bacterial growth curve viz. the lag, log, stationary, and death

phase.

Phases of Growth

1. Lag phase: It is the preparation time for reproduction and increase in cell number

with active macro molecular synthesis like DNA, RNA, various enzymes and other

structural components.

2. Exponential (log) phase: Precedes at a logarithmic rate, and determined by the

medium and condition of the culture.

3. Stationary phase:153 The period when the bacteria have achieved their maximal

cell density or yield. The growth rate is exactly equal to the death rate. During this

phase, the number of viable cells often declines exponentially, with cells dying at a

constant rate.

4. Death phase: The period at which the rate of death of bacterial cells exceeds the

rate of new cell formation. There is drastic decline in viable cells.

Literature review


Gram staining:154 The procedure to differentiate the gram positive and gram

negative microorganism. It involves the application of methylene blue dye, gram’s

iodine, ethanol and safranine respectively. The difference between Gram-positive

and Gram-negative bacteria is in the permeability of the cell wall to these

complexes on treatment with mixtures of acetone and alcohol solvents. Thus, cells

stained purple are Gram positive, and those stained red are Gram negative.

Agar well Diffusion method:155 Inoculum is transferred to agar media and swabbing done

over it. Wells are made with sterile cork bauer on agar media and different concentrations of

aqueous extract is filled in that wells with micropipette. The active phytochemical

constituents diffuse radially outward through the agar, producing a concentration gradient.

A clear zone or ring forms around the wells after incubation if the drug inhibits bacterial

growth. The wider the zone surrounding a well, the pathogen will be more susceptible to

that particular drug. The antibacterial activity of drug is more near to the well and as the

distance from the disk increases, the activity decreases.

AQUEOUS EXTRACTION

Principles and mechanisms156

Soxhlet extraction is a well-established technique except for the extraction of thermo

labile compounds. In this particular method, plant material is placed in a thimble

made up of filter paper, and the funnel filled with condensed fresh solvent from a

distillation flask. When the liquid reaches the overflow level, the solution of the

thimble-holder runs back into the distillation flask with completion of a siphon.

Literature review


Solute is separated from the solvent using distillation in the solvent flask. Solute is

left in the flask and fresh solvent passes back into the plant solid bed. The operation

is repeated until complete extraction is achieved.

Advantages and disadvantages of Soxhlet extraction156

The advantages of Soxhlet extraction include

(1) The displacement of transfer equilibrium by repeatedly bringing fresh solvent into

contact with the solid matrix

(2) Maintaining a relatively high extraction temperature with heat from the distillation

flask

(3) No filtration requirement after leaching and method is very simple and cheap

The main disadvantages of Soxhlet extraction include

(1) A large amount of solvent is used

(2) The large amount of solvent used requires an evaporation/concentration procedure

(3) The possibility of thermal decomposition of the target compounds cannot be

ignored as the extraction usually occurs at the boiling point of the solvent for a long

time. The long time requirement and the requirement of large amounts of solvent

lead to wide.

Methodology

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhidravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference tovranahara karma” Page 64

METHODOLOGY

Methodology of the research work plays an important role as proper following

the steps as per plan, will result in minimal errors and effective results. The proper

and standard protocol for carrying out the procedure should be made and it should be

reviewed repeatedly and then followed. The methodology of this work was classified

into following sections for the sake of convenience.

a) Pharmacognostical study

b) Experimental study

Pharmacognostical Study

Collection of drug

i. Flowers ofWoodfordia fruticosa (L.) Kurz. was collected from Botanical

garden of Shri Dharmasthala Manjunatheshwara (SDM) college of Ayurveda and

Hospital, Hassan during the month of november- december.

ii. The roots of Glycyrrhiza glabra Linn. was collected from Kajarekar Pharmacy,

Belgaum, Karnataka

Authentication of the raw drug

The authentication of root of Yashtimadhu (Glycyrrhiza glabra Linn.) and flower

of Dhataki (Woodfordia fruticosa (L.) Kurz) was done at Department of

Methodology


DravyaGuna, SDM College of Ayurveda and Hospital, Hassan.

Preparation of extract

Aqueous extract of the root of Yashtimadhu (Glycyrrhiza glabra Linn) and flower

of Dhataki (Woodfordia fruticosa (L.) Kurz) was prepared based on API

(Ayurveda Pharmacopoeia of India) standards.

Extraction was done using Soxhlet apparatus.

Ingredients:

Table No – 24: Ingredients for preparation of the extracts

Sl.no Sanskrit

name

Botanical name Parts

used

Quantity used

1 Yashtimadhu

(aqueous

extract)

Glycyrrhiza glabra Linn ROOT 100gm coarse

powder

1000ml distilled

water

2 Dhataki

(aqueous

extract)

Woodfordia fruticosa (L)

Kurz

FLOWER 100gm coarse

powder

1000 ml distilled

water

Methodology


Procedure

Soxhlet extractor / apparatus consists of a round bottom flask holding the

menstrum, the extractor of a cylindrical percolator(body) in the middle provided with

an attached siphon and a reflux condenser, fitted at the top.

The material to be extracted (raw material) in crushed/ powdered form is usually

placed in a thimble (made of filter paper) and then inserted into the extractor. The

menstrum is placed in the round bottom flask and boiled. The vapours arising from

the flask pass by the side tube into condenser. The vapours are condensed and drips

into body of the extractor as pure menstrum. It percolates through the drug to be

extracted dissolving the soluble constituents. As soon as the level of menstrum in the

main extractor rises above the siphon bend, the extract is drained out flowing through

the siphon into the flask. A limited amount of hot solvent is thus made to percolate

repeatedly through the raw material solute from which it is transferred to the falsk.

The process is continuous and can be continued as desired. Materials to be extracted

can be extracted with various solvents in the same apparatus in a hot atmosphere.

During each cycle, a portion of the non- volatile compound dissolves in the solvent.

After many cycles the desired compound is concentrated in the distillation flask. The

advantage of this system is that instead of many portions of warm solvent being

passed though the sample, just one batch of solvent is recycled. After extraction the

solvent is removed, typically by means of a rotatory evaporator, yielding the extracted

Methodology


compound. The non-soluble portion of the extracted solid remains in the thimble, and

is usually discarded

Macroscopy

The external feature of the test samples were documented using Canon IXUS digital

camera. The macroscopic features were compared to local flora for authentication.

Microscopy

Sample was preserved in fixative solution. The fixative used was FAA (Formalin-5ml

+ Acetic acid-5ml + 70% Ethyl alcohol-90ml). The materials were left in FAA for

more than 48 hours. The preserved specimens were cut into thin transverse section

using a sharp blade and the sections were stained with saffranine. The slides were also

stained with iodine in potassium iodide for detection of starch. Transverse sections

were photographed using Zeiss AXIO trinocular microscope attached with Zeiss

AxioCam camera under bright field light. Magnifications of the figures are indicated

by the scale-bars.

Physical evaluation

Loss on drying at 105oC

10 g of sample (powder) as placed in tared evaporating dish. It was dried at 105˚C for

5 hours in hot air oven and weighed. The drying was continued until difference

between two successive weights was not more than 0.01 after cooling in desiccator.

Percentage of moisture was calculated with reference to weight of the sample.

Methodology


Total Ash

2 g of sample was incinerated in a tared platinum crucible at temperature not

exceeding 450˚C until carbon free ash is obtained. Percentage of ash was calculated

with reference to weight of the sample.

Acid insoluble Ash

To the crucible containing total ash, add 25ml of dilute HCl and boil. Collect the

insoluble matter on ashless filter paper (Whatmann 41) and wash with hot water until

the filtrate is neutral. Transfer the filter paper containing the insoluble matter to the

original crucible, dry on a hot plate and ignite to constant weight. Allow the residue to

cool in suitable desiccator for 30 mins and weigh without delay. Calculate the content

of acid insoluble ash with reference to the air dried drug.

Water soluble ash

Boil the ash for 5 min with 25 ml of water; collect insoluble matter on an ashless filter

paper, wash with hot water, and ignite for 15 min at a temperature not exceeding

450˚C. Subtract the weight of the insoluble matter from the weight of the ash; the

difference in weight represents the water soluble ash with reference to the air-dried

sample.

Alcohol soluble extractive

Weigh accurately 4 g of the sample in a glass stoppered flask. Add 100 ml of distilled

Alcohol (approximately 95%). Shake occasionally for 6 hours. Allow to stand for 18

hours. Filter rapidly taking care not to lose any solvent. Pipette out 25ml of the filtrate

in a pre-weighed 100 ml beaker. Evaporate to dryness on a water bath. Keep it in an

Methodology


air oven at 105C for 6 hours, cool in desiccator for 30 minutes and weigh. Calculate

the percentage of Alcohol extractable matter of the sample. Repeat the experiment

twice, and take the average value.

Water soluble extractive:

Weigh accurately 4 g of the sample in a glass stoppered flask. Add 100 ml of distilled

water, shake occasionally for 6 hours. Allow to stand for 18 hours. Filter rapidly

taking care not to lose any solvent. Pipette out 25ml of the filtrate in a pre-weighed

100 ml beaker. Evaporate to dryness on a water bath. Keep it in an air oven at 105C

for 6 hours. Cool in a desiccator and weigh. Repeat the experiment twice. Take the

average value.

Preliminary phytochemical tests

Tests for alkaloids

Dragendroff’s test: To a few mg of extract dissolved in alcohol, a few drops of

acetic acid and Dragendroff’s reagent were added and shaken well. An orange red

precipitate formed indicates the presence of alkaloids.

Wagners’s test: To a few mg of extract dissolved in acetic acid, a few drops of

Wagner’s reagent was added. A reddish brown precipitate formed indicates the

presence of alkaloids.

Mayer’s test: To a few mg of extract dissolved in acetic acid, a few drops of

Mayer’s reagent was added. A dull white precipitate formed indicates the

presence of alkaloids.

Methodology


Hager’s test: To a few mg of extract dissolved in acetic acid, 3 ml of Hager’s

reagent was added, the formation of yellow precipitate indicates the presence of

alkaloids.

Tests for carbohydrates

Molisch’s test: To the extract, 1 ml of α-naphthol solution and conc. sulphuric

acid were added along the sides of test tube. Violet colour formed at the

junction of the two liquids indicates the presence of carbohydrates.

Fehling’s test: A few mg of extract was mixed with equal quantities of

Fehling’s solution A and B. The mixture was warmed on a water bath. The

formation of a brick red precipitate indicates the presence of carbohydrates.

Benedict’s test: To 5 ml of Benedict’s reagent, a few mg of extract was added,

and boiled for two minutes and cooled. Formation of a red precipitate indicates

the presence of carbohydrates.

Test for steroids

Libermann-Burchard test:To the extract was dissolved in chloroform, 1 ml of

acetic acid and 1 ml of acetic anhydride were added, then heated on a water bath

and cooled. Few drops of conc. Sulphuric acid were added along the sides of the

test tube. Appearance of bluish green colour indicates the presence of steroids.

Salkowski test: The extract was dissolved in chloroform and equal volume of

conc. Sulphuric acid was added. Formation of bluish red to cherry red colour in

chloroform layer and green fluorescence in the acid layer indicates the presence

of steroids.

Methodology


Test for saponins

To a few mg of extract, distilled water was added and shaken. Stable froth

formation indicates the presence of saponin.

Test for tannins

To the extract, a few drops of dilute solution of ferric chloride was added,

formation of dark blue colour shows the presence of tannins.

Test for flavonoids

Shinoda’s test: To the extract in alcohol, a few magnesium turnings and few

drops of conc. hydrochloric acid were added and heated on a water bath.

Formation of red to pink colour indicates the presence of flavonoids.

Test for phenol

To the extract in alcohol, added two drops of alcoholic ferric chloride. Formation

of blue to blue black indicates the presence of phenol.

Test for coumarins

To the extract in alcohol,a few drops of 2 N sodium hydroxide solution was

added. Dark yellow colour formation indicates the presence of coumarins.

Test for triterpenoids

The extract was warmed with tin bits and few drops of thionyl chloride.

Formation of pink colour indicates the presence of triterpenoids.

Test for carboxylic acid

Extract dissolved in water is treated with sodium bicarbonate. Brisk effervescence

indicates the presence of carboxylic acid.

Methodology


Test for resin

Few mg of the sample was mixed with water and acetone. Turbidity indicates the

presence of resins.

Test for quinone

A few mg of alcohol extract was treated with 0.5% of sodium hydroxide. Deep

coloration like pink, purple or red indicates the presence of quinone.

HPTLC

1g of each of Yastimadhu root (Glycyrrhiza glabra Linn.) coarse powder and flower

of Dhataki (Woodfordia fruticosa (L.) Kurz.) were extracted with 10 ml of alcohol. 4,

8 and 12µl 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 n-Butanol: Water: Glacial acetic acid (7.0: 2.0: 1.0) for yastimadhu

root and dhataki Toluene: Chloroform: Ethyl Acetate: Formic acid (2.0: 6.0: 6.0: 2.0),

Toluene: Ethyl Acetate: Methanol: Formic acid (3.0: 3.0: 0.8: 0.2). The developed

plates were visualized in short UV, long UV and then derivatised with vanillin

sulphuric acid (post derivatisation under white light). Subsequently scanned under UV

254nm, 366nm. Rf, colour of the spots and densitometric scan were recorded.

Methodology


EXPERIMENTAL STUDY

Source of data

The current study was basically a microbiology based model where minimum of 60

patients fulfilling diagnostic and inclusion criteria was included for study from OPD and

IPD of Sri Dharmasthala Manjunatheshwara College of Ayurveda and Hospital, Hassan

Method of Collection of data

A proforma containing detailed information on each patient was prepared according

to the protocol designed for the study.

Pus Specimen was collected from Dushta Vrana (non healing ulcer) by touching the

infected area with a sterile swab. Transfer of inoculum to MacConkey’s agar and

Blood agar plate. Culturing was done by streak culture method and it was subjected to

microscopic examination for identification of Staphylococcus aureus, Pseudomonas

aeruginosa, Escherichia coli, Acenetobacter bacteria. Further cultured organism was

subjected for subculturing and antibacterial assay was performed with aqueous

extracts of Yashtimadhu and Dhataki adopting Agar well diffusion method.

a) Diagnostic criteria

Patients complaining of Dushta vrana associated with one or more following symptoms

Deerghakaleena

Poothipooya

Methodology


Ateevavedana

Daha

Kandu

Sopha

b) Inclusion criteria

Patients of all age group of either gender fulfilling the diagnostic criteria.

c) Exclusion criteria

Subjects diagnosed with Leprosy, Tuberculosis, Malignancy, HIV, HBsAg positive

subject will be excluded

Subjects with any complications which may interfere with the course of study

Research Design

An experimental study

Methodology:

The pus sample from the patients suffering from Dushta vrana was cultured. The

results showing presence of Staphyloccus aureus, Escherichia coli Pseudomonas

Methodology


aeruginosa, Acenetobacter was further examined for sensitivity with Yashtimadhu

and Dhataki as follows: Patients fulfilling the diagnostic and inclusion criteria was

included in the study. Detailed history was recorded in specially designed case

proforma. Pus sample was collected. Transferred the inoculum to Macconkey agar

plate and culturing was done by Streak culture method and subjected to microscopical

examination for the identification of Staphyloccus aureus, Escherichia coli

Pseudomonas aeruginosa, Acenetobacter bacteria. Then sensitivity test was

performed by Agar well diffusion method.

Collection of pus sample: Pus Specimen was collected from Dushta Vrana (non

healing ulcer) by touching the infected area with a sterile swab.

Culturing bacteria in pus sample: Streak culture method

Requirements: Macconkey agar plate, nichrome loop, gas burner, incubator (37°C).

In order to culture the bacteria, culture media has to be prepared Culture media

preparation

Macconkey Agar preparation

For preparation of 500 ml of Macconkey Agar solid media: 17.5 grams of

Macconkey agar was weighed and mixed with 7.5 grams of Agar Agar.

Later the powder was dissolved in 500 ml of distilled water and transferred to a

conical or flat bottom flask.

Mouth of the flask was sealed with cotton plug covered with a layer of paper and

tightened with bands to avoid any spilling of liquid from within. And was

Methodology


autoclaved at 121°C for 20 minutes.

Muller hinton agar preparation

For preparation of 500 ml of Muller hinton agar solid media: 19.5 grams of

Muller hinton agar was weighed and mixed with 7.5 grams of Agar Agar.

Later the powder was dissolved in 500 ml of distilled water and transferred to a

conical or flat bottom flask.

Mouth of the flask was sealed with cotton plug covered with a layer of paper and

tightened with bands to avoid any spilling of liquid from within. And was

autoclaved at 121°C for 20 minutes.

Procedure for culturing of bacteria:

The outer lower surface of the sterile Macconkey agar plate was marked by using

a glass marking pencil (zone of inoculation).

Red heated the nichrome loop and after cooling it, transfered aseptically, a loop-

full of bacteria to the previously marked area (zone of inoculation) on the

nutrient agar plate and streak it perpendicular to the zone of inoculation.

Placed the plate in the incubator at 37°C and observed the results after 24 hours.

Then for the identification of bacteria microscopical examination was done.

Methodology


Sensitivity test

Sensitivity test was performed in Muller hinton agar

Agar well diffusion method

Requirements: Mullerhinton agar, Aqueous extract of Yashtimadhu, Aqueous

extract of Dhataki, Cork borer

Procedure:

Cleaned the work place in laminar air flow using 70% of Ethyl alcohol and

switched on to UV for 20 min.

Poured around 15 ml Mullerhinton agar media uniformly over the petridish,

mixed well and allowed the media to solidify for 30 minutes.

The agar plate surface is inoculated by spreading a volume of the microbial

inoculam over the entire agar surface.

Maked 6 equidistant wells on the plate with cork borer and added different

concentrations of alcoholic extracts into the wells.

Tests were conducted for 6 different concentrations of extract of Yashtimadhu

and Dhataki (600 µl, 500 µl, 400 µl, 300 µl, 200 µl & 100 µl) separately.

Incubated the petridishes at 37°C for 24 hours. After the incubation period, the

zone of inhibition was measured with a ruler.

Methodology


Assessment Criteria

Sensitivity test was done by cork bourer well diffusion method. Two dishes were

separately used for Yashtimadhu (Glycyrrhiza glabra Linn.) and Dhataki (Woodfordia

fruticosa (L.) Kurz.). Initially 6 wells in each dish was charged with 6 different

concentration of aqueous extracts of Yashtimadhu and Dhataki respectively. If the

drug is sensitive a clear circular “ halo” ( technically known as “ plaque” or Zone of

Inhibition) will appear around the well, indicating absence of bacteria. If that zone

appears, it shows that the particular drug is effective against the specific bacteria

(Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Acinetobacter)

Analytical parameter

The disc diffusion study was measured by following zones.

• Sensitive (S) zone

• Moderate (M)/Intermediate (I) sensitive zone

• Resistant (R) zone.

Statistics

Data was collected using case report format (CRF) designed by incorporating

all aspects (Ayurveda & modern medicine) for the study. Such collected data

was tabulated and analysed using SPSS version 23. by using One Way

ANOVA Test followed by unpaired t - test. Demographic data and other

relevant information were analysed with descriptive statistics. Continuous data

Methodology


was expressed in mean +/- standard deviation, and nominal and ordinal data

was expressed in percentage

Level of significance : P=0.01- 0.001 = statistically highly significant

P=0.01-0.05 = statistically significant

P> 0.05=not significant

Sample Size Estimation

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz.) as pratinidhi dravya forYashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranahara karma”

Page 80

Sample Size Estimation

Not Applicable

OBSERVATIONS AND RESULTS


Page 81

OBSERVATION AND RESULTS

Research work is complete with only accurate recording of observations. The findings of

the work should be noted down with the probable reasoning. Here all the observations

and findings were noted down in each and every step of the work. The observations and

results are enlisted below.

a) Pharmacognostical study

b) Experimental study

PHARMACOGNOSTICAL STUDY

Preparation of extract -

Glycyrrhiza glabra Linn.

During the process of Soxhelt extraction total of 21 siphoning was considered for

the completion of extraction, 1st siphoning occurred after 1hrs of the extraction followed

by other siphoning which occurred with a gap of 10 min consecutively. Siphoning was

continued till the solvent in the siphoning tube appears colourless (which was attained on

21st siphoning). Aqueous extract was pale brown in colour. Extract was liquid in

consistency with pleasant odour.

Woodfordia fruticosa (L.) Kurz

During the process of Soxhelt extraction total of 20 siphoning was considered for

the completion of extraction, 1st siphoning occurred after half hours of the extraction

followed by other siphoning which occurred with a gap of 7-8 min consecutively.



Page 82

Siphoning was continued till the solvent in the siphoning tube appears colourless (which

was attained on 20th siphoning). Aqueous extract was reddish orange in colour. Extract

was liquid in consistency with pleasant odour.

Yashtimadhu (Glycyrrhiza glabra Linn.)

Macroscopic evaluation

Table No – 25: Showing macroscopic evaluation of root of Yashtimadhu


Root of Yashtimadhu (Glycyrrhiza glabra Linn.)

Appearance Cylindrical, longitudinally wrinkled pieces, transversely cut

and smoothened surface

Texture Rough

Colour Yellowish brown

Taste Sweet

Odour Characteristic odour



Page 83

Microscopic evaluation

Root of Glycyrrhiza glabra Linn.

Transverse section

Cork: 10-20 or more layers of tabular cells, outer layers with reddish brown

amorphous contents, inner 3 or 4 rows having thicker, colourless walls

Secondary Cortex: usually of 1-3 layers of radially arranged parenchymatous cells

Phloem: a broad band, parenchymatous cells of inner part cellulosic and outer

lignified; radially arranged groups of about 10 or 50 fibre.

Cambium: form a tissue of 3 or more layers of cells

Secondary xylem: distinctly radiate with medullary rays, 3 or 5 cells wide,

yellow, pitted, reticulately thickened walls, groups of lignified fibres with crystal

sheaths similar to those of phloem

Pith: parenchymatous cells in longitudinal rows, with intercellular spaces

Physiochemical parameters

Table No – 26: Showing Physiochemical parameter of Yashtimadhu

Standard value Obtained value

Foreign matter Not more than 2% absent

Loss on drying Not more than 12% 11.8%



Page 84

Total Ash Not more than 10% 7.5%

Acid Insoluble Ash Not more than 2.5% 1%


value

Not less than 10% 10%

Water soluble extractive value Not less than 20% 31%

Preliminary Phytochemicals:

Table No – 27: Showing results of preliminary phytochemical screening of aqueous

extract

Test Inference

Glycyrrhiza glabra Linn.

(Aqueous extract)

Alkaloid +

Steroid +

Carbohydrate +

Tannin +

Flavanoids +

Saponins +

Tri terpenoid +

Coumarins -

Phenols +

Carboxylic acid +

Amino acids -



Page 85

(+) - present; (-) – negative

HPTLC

Figure 1. HPTLC photo documentation of ethanol extract of Yastimadhu root

Short UV Long UV After derivatisationSolvent system – n-butanol: Water: Glacial acetic acid (7.0: 2.0: 1.0)

Track 1 – Yastimadhu root – 4µlTrack 2 – Yastimadhu root – 8µlTrack 2 – Yastimadhu root – 12µl

Table No - 28: Rf values of sample of Yastimadhu rootSolvent system – n-butanol: Water: Glacial acetic acid (7.0: 2.0: 1.0)

Short UV Long UV After derivatisation0.20 (Purple)

0.35 (L. green) - -- 0.37 (F. blue) -

0.49 (L. green) - -- 0.61 (F. blue) -

0.65 (D. green) 0.65 (F. blue) 0.65 (Yellow)- 0.74 (F. blue) -

0.78 (D. green) 0.78 (FD. blue) 0.78 (Yellow)*D – dark; L – light; F – fluorescent

Resin +

Quinone +



Page 86

In Glycyrrhiza glabra Linn. {Solvent system – n-butanol: Water: Glacial acetic acid (7.0:

2.0: 1.0)] - 8 spots were detected in different Rf value.

Figure 2. Densitometric scan of Yastimadhu root

Fig 2a. At 254nm

Fig 2b. At 366nm



Page 87

Fig 2c. At 620nm

HPTLC Densitometric Scan

Total 17 numbers of active components were detected in Glycyrrhiza glabra Linn.

having Rf value (0.05, 0.44, 0.54, 0.59, 0.75, 0.89, 0.13, 0.20, 0.47, 0.56, 0.72, 0.84, 0.97,

0.24, 0.26, 0.42, 0.75)

Under short UV (254nm) , 6 peaks were detected . Among them maximum percentage

of area were occupied by Rf 0.75 (50.72%), 0.89 (14.88%), 0.44 (12.46%).

Under long UV (366nm), 7 peaks were detected . Among them maximum percentage of

area were occupied by Rf 0.72 (28.79%), 0.84 (15. 71%).

After derivatization (620nm), 5 peaks were detected. Among them maximum

percentage of area were occupied by Rf 0.24 (38.96%) , 0.26 (37.60%)



Page 88

Dhataki (Woodfordia fruticosa (L.) Kurz)Macroscopic evaluation

Table No – 29: Showing macroscopic evaluation of flower of Dhataki (Woodfordia

fruticosa (L.) Kurz)

Microscopic evaluation

Flowers ofWoodfordia fruticosa (L.) Kurz.

Transverse section of stalk shows an epidermis formed by thick wall cells with cuticle; 2

- 3 layers of collenchyma ; cortex formed by 4 - 5 layers of reticulate parenchyma; inner

to cortex continuous ring of phloem followed by xylem. The centre is occupied by pith

formed by pitted parenchyma

Flower of Dhataki (Woodfordia fruticosa (L.) Kurz)

Appearance Flower is about 1.2 cm long, occurs as singles

or in bunches; with campanulate base and

oblique apex; very minute accessory sepals

attached outside at the juncture of calyx tooth

Texture Smooth, Glabrous

Colour Scarlet red in colour

Taste Astringent

Odour Characteristic odour



Page 89

Transervese section of Corolla shows upper and lower epidermis with few covering

trichomes, present of vascular bundle. Beneath the upper epidermis shows layer of

palisade cells. The lower epidermis having spongy parenchyma.

Transverse section of Anther shows epidermis. Underneath the epidermis shows presence

of pollen grains

Transverse section of Ovary shows an epidermis with covering trichome; cortex shows

group of thin walled fibres; rudimentary cells which would form fruit wall and seed wall

are seen. The central portion is formed by thin walled parenchyma which shows pith like

tissue of the axis of the ovary.

Physiochemical parameters

Table No – 30: Showing Physiochemical parameter of Dhataki

Standard value Obtained value

Foreign matter Not more than 2% 0.17%

Loss on drying - 13.1 %

Total Ash Not more than 10% 6%

Acid Insoluble Ash Not more than 0.5% 8%


value

Not less than 8 % 18%

Water soluble extractive value Not less than 20% 41%



Page 90

Preliminary Phytochemicals:

Table No – 31: Showing Results of preliminary phytochemical screening of aqueous

extract

(+) - present; (-) – negative

Test Inference

Woodfordia fruticosa (L.) Kurz

(Aqueous extract)

Alkaloid -

Steroid -

Carbohydrate +

Tannin +

Flavanoids +

Saponins +

Tri terpenoid -

Coumarins -

Phenols +

Carboxylic acid +

Amino acids -

Resin -

Quinone +



Page 91

HPTLC

Figure 3. HPTLC photo documentation of ethanol extract of Dhataki pushpa

Short UV Long UV After derivatisationSolvent system - Toluene: Chloroform: Ethyl Acetate: Formic acid (2.0: 6.0: 6.0: 2.0)

Track 1 – Dhataki pushpa– 4µlTrack 2 – Dhataki pushpa– 8µlTrack 2 – Dhataki pushpa– 12µl

Table No – 32: Rf values of sample of Dhataki pushpaSolvent system - Toluene: Chloroform: Ethyl Acetate: Formic acid (2.0: 6.0: 6.0: 2.0)

Short UV Long UV After derivatisation0.08 (Green) - -

- 0.06 (F. blue) -- - 0.21 (Purple)

0.32 (Green) - -0.42 (Green) - -

- - 0.44 (Purple)- - 0.57 (Purple)- 0.65 (F. blue) -

0.70 (Green) 0.70 (F. red) 0.71 (Purple)0.78 (Green) - -

- 0.81 (F. red) -0.84 (Green) 0.84 (F. blue) -

- 0.87 (F. blue) -*D – dark; L – light; F – fluorescent



Page 92

In Woodfordia fruticosa (L.) Kurz [Solvent system - Toluene: Chloroform: Ethyl Acetate:

Formic acid (2.0: 6.0: 6.0: 2.0)] - 13 spots were detected in different Rf value.

Figure 4. Densitometric scan of Dhataki pushpaSolvent system - Toluene: Chloroform: Ethyl Acetate: Formic acid (2.0: 6.0: 6.0: 2.0)

Fig 4a. At 254nm



Page 93

Fig 4c. At 620nm

Fig 4b. At 366nm



Page 94

HPTLC Densitometric scan

Total 26 number of active components were detected in Woodfordia fruticosa (L.) Kurz

having Rf value (0.05, 0.15, 0.21, 0.41, 0.52, 0.68, 0.82, 0.92, 0.98, 0.03, 0.06, 0.11, 0.24,

0.35, 0.46, 0.72, 0.76, 0.93, 0.07, 0.28, 0.42, 0.53, 0.69, 0.80, 0.83, 0.94)

Under short UV (254 nm) , 9 peaks were detected. Among them maximum percentage

of area were occupied by Rf 0.05 (58.24%), 0.15 (11.15%)

Under long UV (366 nm) , 10 peaks were detected. Among them maximum percentage

of area were occupied by Rf 0.93 (33.38%) , 0.11 (15.37%), 0.82 (15.32%)

After derivatization (620 nm), 9 peaks were detected. Among them maximun

percentage of area were occupied by Rf 0.07 (60.28%), 0.53 (11.57%)

Figure 5. HPTLC photo documentation of ethanol extract of Dhataki pushpa

Short UV Long UV After derivatisationSolvent system - Toluene: Ethyl Acetate: Methanol: Formic acid (3.0: 3.0: 0.8: 0.2)

Track 1 – Dhataki pushpa– 4µlTrack 2 – Dhataki pushpa– 8µlTrack 2 – Dhataki pushpa– 12µl



Page 95

Table No - 33: Rf values of sample of Dhataki pushpaSolvent system - Toluene: Ethyl Acetate: Methanol: Formic acid (3.0: 3.0: 0.8: 0.2)

Short UV Long UV After derivatisation0.25 (Green) - -

- - 0.27 (L. purple)- - 0.41 (L. purple)- 0.43 (F. blue) -- 0.48 (F. red) -

0.53 (Green) - 0.53 (L. purple)- 0.56 (F. red) -- 0.64 (F. blue) -- 0.72 (F. blue) -

0.75 (Green) - -- - 0.79 (D. purple)- 0.89 (F. blue) -

*D – dark; L – light; F – fluorescent

.In Woodfordia fruticosa (L.) Kurz [Solvent system - Toluene: Ethyl Acetate: Methanol:

Formic acid (3.0: 3.0: 0.8: 0.2)] - 12 spots were detected in different Rf value.

Figure 6. Densitometric scan of Dhataki pushpa

Solvent system - Toluene: Ethyl Acetate: Methanol: Formic acid (3.0: 3.0: 0.8: 0.2)



Page 96

Fig 6a. At 254nm

Fig 6b. At 366nm



Page 97

Fig 6c. At 620nm

HPTLC Densitometric Scan

Total 25 number of active components were detected in Woodfordia fruticosa (L.) Kurz

having Rf value (0.05, 0.32, 0.43, 0.58, 0.63, 0.66, 0.87, 0.04, 0.06, 0.20, 0.23, 0.27, 0.39,

0.50, 0.53, 0.55, 0.62, 0.67, 0.76, 0.85, 0.92, 0.10, 0.34, 0.49, 0.93)

Under short UV (254nm), 7 peaks were detected. Among them maximum percentage of

area were occupied by Rf 0.05 (68.70%), 0.32 (18.27%)

Under long UV (366nm), 15 peaks were detected. Among them maximum percentage of

area were occupied by Rf 0.67 (17.14%), 0.58 (10.35%), 0.55 (8.82%)

After derivatization (620nm), 7 peaks were detected. Among them maximum

percentage of area were occupied by Rf 0.06 (38.40%), 0.05 (38.32%)



Page 98

EXPERIMENTAL STUDY

In vitro study

In the present study, 60 patients of Dushta vrana were selected on the basis of

diagnostic and inclusion criteria and their pus sample was collected and culturing was

done by Streak culture method and subjected to microscopical examination for the

identification of Staphyloccus aureus, Escherichia coli, Pseudomonas aeruginosa,

Acenetobacter and the same has been subjected to antibacterial assay with 6 different

concentrations of aqueous extract of both Yashtimadhu and Dhataki by agar well

diffusion method. From the data it is evident that the aqueous extracts of Yashtimadhu

and Dhataki showed antimicrobial activity against Staphylococcus aureus, Escherichia

coli, Acinetobacter, Pseudomonas aeruginosa .

Table No – 34: Number of patients

No. of patients Percentage

Included 60 75

Excluded 20 25

Total (Screened) 80 100

Out of 80 patients screened , 60 patients was included and remaining 20 patients was

excluded .



Page 99

Table No – 35: Number of organisms

Organisms No.of samples Percentage

Staphylococcus aureus 25 41.7 %

Escherichia coli 18 30.0 %

Pseudomonas aeruginosa 8 13.3%

Acinetobacter 9 15.0%

Total 60 100 %

Among 60 pus samples 25 were identified as Staphylococcus aureus organisms (41.7%),

18 identified as Escherichia coli (30%), 9 were identified as Acinetobacter (15%) and 8

were identified as Pseudomonas aeruginosa (13.3%).



Page 100


Table No – 36: Colony Characterization and Microscopy of Staphylococcus aureus

Observation on Antibacterial activity shown at different concentrations of aqueous

extract of Yashtimadhu against Staphylococcus aureus

Table No - 37 : Distribution based on sensitivity of aqueous extracts of different

concentrations of Yashtimadhu against Staphylococcus aureus

Sample Incubation

Period

Growth onMacCONKE

YAgar

CultureCharacters

MicroscopyBy Gram’sstaining

Organismidentified

Urine 37◦C for24 hrs

Growth seen

Size(in mm)

1- 2 mm

Grampositive

Staphylococcusaureus

Shape Circular

Surface Smooth

Elevation Convex

Edge Entire

Opacity Opaque

Colour ofColony

Pink

Consistency Buttery

Extract *ZOI in mmagainst


600 µg/µlN=25

500 µg/µlN=25

400 µg/µlN=25

300 µg/µlN=25

200 µg/µlN=25

100 µg/µlN=25

F % F % F % F % F % F %

0 8 32 9 36 10 40 10 40 10 40 14 56



Page 101

* ZOI = Zone Of Inhibition* F = Frequency of samples showing sensitivity against aqueous extract of Yashtimadhu* N = Total no. of samples

Sensitivity of aqueous extract of different concentrations of Yashtimadhu (600, 500, 400, 300,

200, 100 µg/µl) against Staphylococcus aureus in 25 samples isolated by culture and

sensitivity has drawn the following data.

At 600 µg/µl Yashtimadhu showed maximum zone of inhibition was 28 mm in 1 (4%) sample,

25 mm in 1 (4%), 21 mm in 1 sample (4%), 18 mm in 4 (16%) samples, 17 mm in 2 (8%)

samples, 16 mm in 2 (8%) samples, 15 mm in 3 (12%) samples, 12 mm in 1 sample (4%) and

Aqueousextract ofYashti-madhu

7 1 4

8 1 4

10 2 8 3 12 3 12 3 12 3 12 2 8

11 1 4 1 4

12 1 4 1 4 2 8 2 8

13 3 12 1 4 3 12

14 2 8 1 4 1 4

15 3 12 5 20 2 8 2 8 4 16 1 4

16 2 8 1 4 2 8 1 4 2 8

17 2 8 2 8 1 4 3 12

18 4 16 2 8 1 4 1 4 1 4

19 2 8

20 1 4 1 4 1 4 1 4

21 1 4

25 1 4 1 4

28 1 4



Page 102

minimum zone of inhibition 10 mm was observed in 2 (8%) samples and there was no zone of

inhibition observed in 8 (32%) samples.

At 500 µg/µl, it showed that maximum zone of inhibition was 25 mm in 1 (4%) sample, 20

mm in 1 sample (4%), 18 mm in 2 (8%) samples, 17 mm in 2 (8%) samples, 16 mm in 1

sample (4%), 15 mm in 5 samples (20%), 10 mm in 3 samples (12%) and minimum zone of

inhibition 8 mm was observed in 1 (4%) sample and there was no zone of inhibition observed

in 9 (36%) samples.


mm in 2 (8%) samples, 18 mm in 1 (4%) samples, 17 mm in 1 sample (4%), 16 mm in 2

samples (8%), 15 mm in 2 (8%) samples, 13 mm in 3 (12%) samples and minimum zone of

inhibition 10 mm was observed in 3 (12%) samples and there was no zone of inhibition

observed in 10 (40%) samples.


mm in 1 (4%) sample, 17 mm in 3 (12%) samples, 16 mm in 1 (4%) sample, 15 mm in 2 (8%)

samples, 14 mm in 2 (8%) samples, 13mm in 1 (4%) sample,12 mm in 1(4%) sample and

minimum zone of inhibition 10 mm was observed in 3 (12%) samples and there was no zone

of inhibition observed in 10 (40%) samples.


mm in 1 (4%) sample, 16 mm in 2 (8%) samples, 15 mm in 4 (16%) samples, 14 mm in 1 (4%)

sample, 12 mm in 2 (8%) samples, 11 mm in 1(4%) sample and minimum zone of inhibition

10 mm was observed in 3 (12%) samples and there was no zone of inhibition observed in 10

(40%) samples.



sample, 10 mm in 2 (8%) samples and minimum zone of inhibition 7 mm was observed in 1

(4%) sample and there was no zone of inhibition observed in 14 (56%) samples.



Page 103


extract of Dhataki against Staphylococcus aureus

Table No – 38: Distribution based on sensitivity of aqueous extracts of different

concentrations of Dhataki against Staphylococcus aureus

Extract*ZOI in mm

against

Staphylococcus

aureus

S

600 µg/µl

N=25

500 µg/µl

N=25

400 µg/µl

N=25

300 µg/µl

N=25

200 µg/µl

N=25

100 µg/µl

N=25

F % F % F % F % F % F %

Aaa Aqueous

extract

of

Dhataki

10 1 4 4 16

11 1 4

12 1 4 3 12 4 16

13 1 4 5 20 3 12 5 20

14 1 4 1 4 2 8

15 2 8 2 8 9 36 2 8 4 16 4 16

16 1 4 3 12

17 2 8 3 12 2 8

18 2 8 1 4 2 8 1 4 1 4

20 5 20 7 28 4 16 6 24 2 8 2 8

21 1 4 1 4

22 3 12 3 12 2 8 2 8 2 8 3 12

23 3 12 2 8 1 4

24 1 4 1 4 2 8 2 8 1 4 1 4



Page 104

* ZOI = Zone Of Inhibition* F = Frequency of samples showing sensitivity against aqueous extract of Dhataki* N = Total no. of samples

Sensitivity of aqueous extract of different concentrations of Dhataki (600, 500, 400, 300, 200,

100 µg/µl) against Staphylococcus aureus in 25 samples isolated by culture and sensitivity

has drawn the following data.

At 600 µg/µl Dhataki showed maximum zone of inhibition was 27 mm in 1 (4%) sample, 26

mm in 1 (4%) sample, 25 mm in 4 samples (16%), 24 mm in 1 (4%) sample, 23 mm in 3(12%)

samples, 22 mm in 3 (12%) samples, 20 mm in 5 (20%) samples, 18 mm in 2 samples (8%),

17 mm in 2 (8%) samples, 15 mm in 2 (8%) samples and minimum zone of inhibition 14 mm

was observed in 1 (4%) sample.


mm in 3 samples (12%), 24 mm in 1 (4%) sample, 23 mm in 2 (8%) samples, 22 mm in 3

samples (12%), 21 mm in 1 sample (4%), 20 mm in 7 samples (28%), 18 mm in 1 (4%)

sample, 17 mm in 3 (12%) samples, 15 mm in 2 (8%) samples and minimum zone of

inhibition 13 mm was observed in 1 (4%) sample.


mm in 1 (4%) sample, 26 mm in 1 (4%) sample, 24 mm in 2 samples (8%), 22 mm in 2

samples (8%), 21 mm in 1 (4%) samples, 20 mm in 4 (16%) samples, 18 mm in 2 (8%)

25 4 16 3 12

26 1 4 1 4 1 4 1 4 1 4

27 1 4 1 4

28 1 4 1 4 1 4

30 1 4 1 4



Page 105

samples, 15 mm in 9 (36%) samples and minimum zone of inhibition 14 mm was observed in

1 (4%) sample.


mm in 1 (4%) sample, 24 mm in 2 (8%) samples, 23 mm in 1 (4%) sample, 22 mm in 2 (8%)

samples, 20 mm in 6 (24%) samples, 18 mm in 1 (4%) sample,16 mm in 1(4%) sample, 15

mm in 2 (8%) samples, 14 mm in 2 (8%) samples, 13 mm in 5 (20%) samples and minimum

zone of inhibition 12 mm was observed in 1 (4%) sample.


mm in 1 (4%) sample, 26 mm in 1 (4%) sample, 24 mm in 1 (4%) sample, 22 mm in 2 (8%)

samples, 20 mm in 2 (8%) samples, 17 mm in 2 (8%) samples, 16 mm in 3 (12%) samples, 15

mm in 4 (16%) samples, 13 mm in 3 (12%) samples, 12 mm in 3 (12%) samples, 11 mm in 1

(4%) sample and minimum zone of inhibition 10 mm was observed in 1 (4%) sample.



samples, 15 mm in 4 (16%) samples, 13 mm in 5 (20%) samples, 12 mm in 4 (16%) samples

and minimum zone of inhibition 10 mm was observed in 4 (16%) samples.

Table No – 39: Comparing antibacterial action of aqueous extracts of Yashtimadhu

and Dhataki against Staphylococcus aureus

Different concentrationsof alcoholic extract

600 µg/µl 500 µg/µl 400 µg/µl 300µg/µl 200 µg/µl 100 µg/µl

N (total samples) 25 25 25 25 25 25Mean of zone of inhibitionOf different concentrationsof aqueous extractof Yashtimadhu (mm)

11.56 9.76 8.96 8.72 8.36 5.20



Page 106

On comparing the antibacterial action of aqueous extracts of Yashtimadhu and Dhataki, the

mean difference between the zone of inhibitions of alcoholic extracts of Yashtimadhu and

Dhataki in different concentrations (600 µg/µl to 100 µg/µl) are 9.48 mm, 10.64 mm, 10.24

mm, 9.6 mm, 8.84 mm and 10.36 mm respectively.

Table No – 40: Comparing the zone of inhibition of aqueous extracts of Yashtimadhu and

Dhataki against Staphylococcus aureus

On comparing the zone of inhibitions of aqueous extracts of Yashtimadhu and Dhataki, Dhataki

had shown more sensitive zones in all the concentrations than Yashtimadhu, thus it is clear that

Dhataki is having better antibacterial action against Staphylococcus aureus than Dhataki. For

different concentrations of both Yashtimadhu and Dhataki, it is evident that increasing the

Mean of zone of inhibitionof different concentrationsof aqueous extractof Dhataki (mm)

21.04 20.40 19.20 18.32 17.20 15.56

Difference of mean in mm9.48 10.64 10.24 9.6 8.84 10.36

Concentrations 600 µg/µl 500 µg/µl 400 µg/µl 300 µg/µl 200 µg/µl 100 µg/µl

S M R S M R S M R S M R S M R S M RNo. of samples(Yashtimadhu)

14 3 8 12 2 11 9 6 10 8 7 10 8 7 10 8 2 15

No. of samples(Dhataki)

14 10 1 11 13 1 8 16 1 7 10 8 6 11 8 5 7 13



Page 107

concentrations significant increase in zone of inhibition. Consecutively number of sensitive

zones increasing and number of resistant zones decreasing when increasing the concentrations.

Here S – Sensitive, M- Moderately sensitive, R- Resistant.

STATISTICS

Statistically analysed with SPSS Version- 23

Results were analysed after Statistical analysis using the One Way Anova Test followed by

Unpaired t-test

Table No – 41: Statistical values of One-Way ANOVA test

Group Conc N Mean SD F value P value Remarks

Staphylo-

coccus

aureus

Yashtimadhu 600 µg/ µl 25 11.56 8.931 1.812 .114 NS

500 µg/ µl 25 9.76 8.202

400 µg/ µl 25 8.96 7.898

300 µg/ µl 25 8.72 7.640

200 µg/ µl 25 8.36 7.342

100 µg/ µl 25 5.20 6.158



Page 108

P< 0.05 - Significant; P< 0.01 - Highly significant

NS - Not significant; HS - Highly significant

Comparing between Yashtimadhu and Dhataki by One way Anova, Yashtimadhu has P value

0.114 which is not statistically significant . Dhataki has P value 0.000 which is highly

significant. Further on comparing the mean values, mean value of Dhataki was greater than

mean value of Yashtimadhu in all the different concentrations.

Table No – 42: Statistical values of unpaired t- test

Dhataki 600 µg/ µl 25 21.04 3.680 5.107 .000 HS

500 µg/ µl 25 20.40 3.440

400 µg/ µl 25 19.20 4.472

300 µg/ µl 25 18.32 4.697

200 µg/ µl 25 17.20 5.454

100 µg/ µl 25 15.56 5.017

Conc Group N Mean SD t test

value

P Remarks

Staphylo-

coccus

aureus

600 µg/ µl Yashtimadhu 25 11.56 8.931 - 4.907 .000 HS

Dhataki 25 21.04 3.680

500 µg/ µl Yashtimadhu 25 9.76 8.202 - 5. 981 .000 HS



Page 109


HS - Highly significant

Comparing the same concentration of aqueous extract of Yashtimadhu and Dhataki by

unpaired t- test it was observed that in all the concentration were found to be statistically

highly significant

Dhataki 25 20.40 3.440


Dhataki 25 19.20 4.472

300 µg/ µl Yashtimadh 25 8.72 7.640 - 5. 352 .000 HS

Dhataki 25 18.32 4.697


Dhataki 25 17.20 5.454


Dhataki 25 15.56 5.017



Page 110

Escherichia coli

Table No – 43: Colony Characterization and Microscopy of Escherichia coli

Sample IncubationPeriod

Growth onMacCONKEY

Agar

CultureCharacters


Organismidentified


Growth seen

Size(in mm)

1- 2mm

Gramnegative

Escherichiacoli

Shape Circular

Surface Smooth

Elevation LowConvex

Edge Entire

Opacity Opaque

Colour ofColony

MoistPink



Page 111


extract of Yashtimadhu against Escherichia coli


concentrations of Yashtimadhu against Escherichia coli


Extract *ZOI in mm

against

Escherichia coli

S

600 µg/µl

N= 18

500 µg/µl

N= 18

400 µg/µl

N= 18

300 µg/µl

N= 18

200 µg/µl

N= 18

100 µg/µl

N= 18

F % F % F % F % F % F %

Aqueous

extract of

Yashtimadhu

0 17 94.4 17 94.4 15 83.3 17 94.4 14 77.8 14 77.8

7 1 5.6

10 1 5.6 1 5.6 1 5.6 2 11.1

11 1 5.6

12 1 5.6 1 5.6

13 1 5.6

15 1 5.6 1 5.6

25 1 5.6

35 1 5.6



Page 112


200, 100 µg/µl) against Escherichia coli in 18 samples isolated by culture and sensitivity has

drawn the following data.

At 600 µg/µl Yashtimadhu showed zone of inhibition was 12 mm in 1 (5.6%) sample and

there was no zone of inhibition observed in 17 (94.4%) samples.

At 500 µg/µl, it showed that zone of inhibition was 10 mm in 1 (5.6%) sample there was no

zone of inhibition observed in 17 (94.4%) samples.

At 400 µg/µl, it showed that maximum zone of inhibition was 35 mm in 1 (5.6%) samples, 13

mm in 1 (5.6%) sample, 10 mm in 1 (5.6%) sample and there was no zone of inhibition

observed in 15 (83.3%) samples.

At 300 µg/µl, it showed zone of inhibition was 11 mm in 1 (5.6%) sample and there was no


At 200 µg/µl, it showed that maximum zone of inhibition was 15 mm in 1 (5.6%) sample, 12

mm in 1 (5.6%) sample, 10 mm in 1 (5.6%) sample, 7 mm in 1 (5.6%) sample and there was

no zone of inhibition observed in 14 (77.8%) samples.


mm in 1 (5.6%) sample, 10 mm in 2 (11.1%) samples and there was no zone of inhibition




Page 113


extract of Dhataki against Escherichia coli


concentrations of Dhataki against Escherichia coli

Extract *ZOI in mm

against

Escherichia coli

S

600 µg/µl

N=18

500 µg/µl

N= 18

400 µg/µl

N= 18

300 µg/µl

N= 18

200 µg/µl

N= 18

100 µg/µl

N= 18

F % F % F % F % F % F %

Aqu Aqueous

extract of

Dhataki

10 2 11.1 2 11.1

11 1 5.6

12 1 5.6 1 5.6 1 5.6 6 33.3

13 1 5.6 1 5.6 3 16.7

14 1 5.6 1 5.6 1 5.6

15 2 11.1 2 11.1 3 16.7 4 22.2 3 16.7 3 16.7

16 2 11.1 2 11.1 1 5.6 1 5.6

17 2 11.1 1 5.6 2 11.1

18 3 16.7 3 16.7 2 11.1 1 5.6

19 1 5.6 2 11.1

20 5 27.8 7 38.9 3 16.7 3 16.7 2 11.1



Page 114



100 µg/µl) against Escherichia coli in 18 samples isolated by culture and sensitivity has


At 600 µg/µl Dhataki showed maximum zone of inhibition was 29 mm in 1 (5.6%) sample, 25

mm in 3 (16.7%) samples, 22 mm in 2 samples (11.1%), 21 mm in 1 (5.6%) sample, 20 mm in

5(27.8%) samples, 18 mm in 3 (16.7%) samples, 15 mm in 2 (11.1%) samples and minimum

zone of inhibition 12 mm was observed in 1 (5.6%) sample.


mm in 1 sample (5.6%), 23 mm in 1 (5.6%) sample, 22 mm in 2 (11.1%) samples, 20 mm in 7

samples (38.9%), 17 mm in 2 samples (11.1%), 15 mm in 2 samples (11.1%), 13 mm in 1

(5.6%) sample and minimum zone of inhibition 12 mm was observed in 1 (5.6%) sample.

21 1 5.6 1 5.6 1 5.6 1 5.6

22 2 11.1 2 11.1 1 5.6 1 5.6

23 1 5.6

24 1 5.6 1 5.6

25 3 16.7 1 5.6 1 5.6 1 5.6 1 5.6

26 1 5.6 1 5.6

28 1 5.6

29 1 5.6 1 5.6 1 5.6



Page 115


mm in 1 (5.6%) sample, 25 mm in 1 (5.6%) sample, 22 mm in 1 sample (5.6%), 21 mm in 1

sample (5.6%), 20 mm in 3 (16.7%) samples, 19 mm in 1 (5.6%) sample, 18 mm in 3 (16.7%)

samples, 17 mm in 1 (5.6%) sample, 16 mm in 2 (11.1%) samples and minimum zone of

inhibition 15 mm was observed in 3 (16.7%) samples.


mm in 1 (5.6%) sample, 21 mm in 1 (5.6%) sample, 20 mm in 3 (16.7%) samples, 18 mm in 2

(11.1%) samples, 17 mm in 2 (11.1%) samples, 16 mm in 2 (11.1%) samples,15 mm in

4(22.2%) samples, 14 mm in 1 (5.6%) sample and minimum zone of inhibition 13 mm was

observed in 1 (5.6%) sample.


mm in 1 (5.6%) sample, 24 mm in 1 (5.6%) sample, 21 mm in 1 (5.6%) sample, 20 mm in 2

(11.1%) samples, 18 mm in 1 (5.6%) sample, 16 mm in 1 (5.6%) sample, 15 mm in 3 (16.7%)

samples, 14 mm in 1 (5.6%) samples, 13 mm in 3 (16.7%) samples, 12 mm in 1 (5.6%)

sample and minimum zone of inhibition 10 mm was observed in 2 (11.1%) samples.


mm in 1 (5.6%) sample, 19 mm in 2 (11.1%) samples, 16 mm in 1 (5.6%) sample, 15 mm in 3

(16.7%) samples, 14 mm in 1 (5.6%) sample, 12 mm in 6 (33.3%) samples, 11 mm in 1 (5.6%)

sample and minimum zone of inhibition 10 mm was observed in 2 (11.1%) samples.



Page 116

Table No – 46: Comparing antibacterial action of aqueous extracts of Yashtimadhuand Dhataki against Escherichia coli


mean difference between the zone of inhibitions of alcoholic extracts of Yashtimadhu and




Dhataki against Escherichia coli




0.67 0.56 3.22 0.61 2.44 3.33


20.33 19.44 19.39 18 16.67 14.56

Difference of mean in mm 19.66 18.88 16.17 17.39 14.23 11.23



1 0 17 0 1 17 2 1 15 1 0 17 2 2 14 2 2 14


12 5 1 12 5 1 10 8 0 7 11 0 6 9 3 4 5 9



Page 117



Dhataki is having better antibacterial action against Escherichia coli than Dhataki. For

different concentrations of Yashtimadhu it is showed that increase the concentrations there is

decrease in zone of inhibition. For different concentrations of Dhataki, it is evident that

increasing the concentrations increase in zone of inhibition.


STATISTICS



unpaired t- test


E. coli Yashtimadhu 600 µg/µl 18 0.67 2.828 1.139 .345 NS

500 µg/µl 18 0.56 2.357

400 µg/µl 18 3.22 8.769

300 µg/µl 18 0.61 2.593

200 µg/µl 18 2.44 4.914

100 µg/µl 18 3.33 7.071

Table No - 48: Statistical values of One-Way ANOVA test



Page 118


NS - Not significant; S - significant


0.345 which is not statistically significant . Dhataki has P value 0.001 which is significant.

Futher on comparing the mean values, the mean value of Dhataki was greater than the mean

value of Yashtimadhu in all the concentrations

Dhataki 600 µg/µl 18 20.33 4.256 4.612 .001 HS

500 µg/µl 18 19.44 4.190

400 µg/µl 18 19.39 3.852

300 µg/µl 18 18.00 4.044

200 µg/µl 18 16.67 4.971

100 µg/µl 18 14.56 4.062



Page 119



Comparing the same concentration of aqueous extract of Yashtimadhu and Dhataki by unpaired

t- test it was observed that in all the concentration were found to be statistically highly

significant.

Conc Group N Mean SD t test valueP Remarks

E.coli 600 µg/µl Yashtimadhu 18 0.67 2.828 -16.327 .000 HS

Dhataki 18 20.33 4.256

500 µg/µl Yashtimadhu 18 0.56 2.357 -16.670 .000 HS

Dhataki 18 19.44 4.190


Dhataki 18 19.39 3.852

300 µg/µl Yashtimadh 18 0.61 2.593 -15.358 .000 HS

Dhataki 18 18 4.044


Dhataki 18 16.67 4.971

100 µg/µl Yashtimadhu 18 3.33 7.071 - 5.839 .000 HS

Dhataki 18 14.56 4.062

Table No – 49: Statistical values of Unpaired t- test



Page 120

Acinetobacter

Table No – 50: Colony Characterization and Microscopy of Acinetobacter


Growth onMacCONKE

YAgar

CultureCharacters


Organismidentified


Growth seen

Size(in mm)

1- 2 mm

Gramnegative

AcinetobacterShape Domed

Surface Smooth

Colour ofColony

Non-pigmented

Consistency Mucoid



Page 121


extract of Yashtimadhu against Acinetobacter


concentrations of Yashtimadhu against Acinetobacter


Extract *ZOI in mm

against

Acinetobacter

S

600 µg/µl

N= 9

500 µg/µl

N= 9

400 µg/µl

N= 9

300 µg/µl

N= 9

200 µg/µl

N= 9

100 µg/µl

N= 9

F % F % F % F % F % F %

Aqueous

extract of

Yashtimadhu

0 5 55.6 5 55.6 6 66.7 8 88.9 8 88.9 9 100

10 1 11.1 1 11.1 2 22.2 1 11.1 1 11.1

13 1 11.1 1 11.1

15 2 22.2 1 11.1

18 1 11.1

20 1 11.1



Page 122


200, 100 µg/µl) against Acinetobacter in 9 samples isolated by culture and sensitivity has


At 600 µg/µl Yashtimadhu showed maximum zone of inhibition was 20 mm in 1 (11.1%)

sample, 18mm in 1 (11.1%) sample, 13mm in 1 (11.1%) sample and minimum zone of

inhibition 10 mm was observed in 1(11.1%) sample and there was no zone of inhibition


At 500 µg/µl, it showed that maximum zone of inhibition was 15 mm in 2 (22.2%) samples,

13mm in 1(11.1%) sample and minimum zone of inhibition 10mm was observed in 1(11.1%)

sample and there was no zone of inhibition observed in 5 (55.6%) samples.

At 400 µg/µl, it showed zone of inhibition was 15 mm in 1 (11.1%) sample, 10 mm in 2

(22.2%) samples and there was no zone of inhibition observed in 6 (66.7%) samples.





At 100 µg/µl, it was observed that there was no zone of inhibition in 9 (100%) samples.



Page 123


extract of Dhataki against Acinetobacter


concentrations of Dhataki against Acinetobacter

Extract *ZOI in mm

against

Acinetobacter

S

600 µg/µl

N= 9

500 µg/µl

N= 9

400 µg/µl

N= 9

300 µg/µl

N= 9

200 µg/µl

N= 9

100 µg/µl

N= 9

F % F % F % F % F % F %

AA Aqueous

extr extract of

Dha Dhataki

10 2 22.2

11 2 22.2

12 1 11.1 3 33.3

13 1 11.1 2 22.2

14 1 11.1 1 11.1 1 11.1

15 3 33.3 3 `33.3 2 22.2 1 11.1

16 1 11.1 1 11.1

17 2 22.2 111111.1 1 11.1 1 11.1

18 3 33.3 2 22.2 2 22.2

20 1 11.1 2 22.2 1 11.1 1 11.1 1 11.1

21 1 11.1



Page 124



100 µg/µl) against Acinetobacter in 9 samples isolated by culture and sensitivity has drawn

the following data.

At 600 µg/µl Dhataki showed maximum zone of inhibition was 25 mm in 2 (22.2%) samples,

23 mm in 1 (11.1%) sample, 22 mm in 1 sample (11.1%), 20 mm in 1 (11.1%) sample, 18 mm

in 3(33.3%) samples and minimum zone of inhibition 13 mm was observed in 1 (11.1%)

sample.


23 mm in 1 sample (11.1%), 20 mm in 2 (22.2%) samples, 18 mm in 2 (22.2%) samples and

minimum zone of inhibition 17 mm was observed in 2 (22.2%) samples.


mm in 1 (11.1%) sample, 21 mm in 1 (11.1%) sample, 20 mm in 1 sample (11.1%), 17 mm in

1 sample (11.1%), 16 mm in 1 (11.1%) sample and minimum zone of inhibition 15 mm was


At 300 µg/µl, it showed that maximum zone of inhibition was 23 mm in 1 (11.1%) sample,

20 mm in 1 (11.1%) sample, 18 mm in 2 (22.2%) samples, 17 mm in 1 (11.1%) sample, 15

22 1 11.1 1 11.1

23 1 11.1 1 11.1 1 11.1 1 11.1

25 2 22.2 2 22.2



Page 125

mm in 3 (33.3%) samples and minimum zone of inhibition 14 mm was observed in 1 (11.1%)

sample.


17 mm in 1 (11.1%) sample, 16 mm in 1 (11.1%) sample, 15 mm in 2 (22.2%) samples, 14

mm in 1 (11.1%) sample, 13 mm in 2 (22.2%) samples and minimum zone of inhibition 12

mm was observed in 1 (11.1%) sample.


mm in 1 (11.1%) sample, 12 mm in 3 (33.3%) samples, 11 mm in 2 (22.2%) samples and


Table No – 53: Comparing antibacterial action of aqueous extracts of Yashtimadhuand Dhataki against Acinetobacter


mean difference between the zone of inhibitions of aqueous extracts of Yashtimadhu and




6.78 5.89 3.89 1.11 1.11 0.00


20.22 20.33 18.22 17.22 15.00 11.89




Page 126




Dhataki against Acinetobacter



Dhataki is having better antibacterial action against Acinetobacter than Dhataki. For different

concentrations of both Yashtimadhu and Dhataki, it is evident that increasing the concentrations

significant increase in zone of inhibition. Consecutively number of sensitive zones increasing

and number of resistant zones decreasing when increasing the concentrations.


Concentrations 600 µg/µl 500 µg/µl 400 µg/µl 300 µg/µl 200 µg/µl 100 µg/µlS M R S M R S M R S M R S M R S M R

No. of samples(Yashtimadhu)

3 1 5 3 1 5 1 2 6 0 1 8 0 1 8 0 0 9


5 4 0 5 4 0 4 5 0 2 7 0 1 7 1 0 2 7



Page 127

STATISTICS



Unpaired T Test



S - Significant; HS - Highly significant


Acineto-

bacter

Yashtimadhu 600 µg/µl 9 6.78 8.511 2.345 0.055 S

500 µg/µl 9 5.89 7.132

400 µg/µl 9 3.89 6.009

300 µg/µl 9 1.11 3.333

200 µg/µl 9 1.11 3.333

100 µg/µl 9 0.00 0.000

Dhataki 600 µg/µl 9 20.22 3.930 10.617 0.000 HS

500 µg/µl 9 20.33 3.240

400 µg/µl 9 18.22 3.270

300 µg/µl 9 17.22 2.906

200 µg/µl 9 15.00 2.449

100 µg/µl 9 11.89 1.691



Page 128


0.055 which is statistically significant . Dhataki has P value 0.000 which is highly significant.

Futher on comparing the mean values, the mean value of Dhataki was greater than the mean

value of Yashtimadhu in all the concentrations.

Table No – 56: Statistical values of unpaired t test


Conc Group N Mean SD t test value P Remarks

Acineto

bacter

600 µg/µl Yashtimadhu 9 6.78 8.511 - 4.302 0.001 HS

Dhataki 9 20.22 3.930


Dhataki 9 20.33 3.240


Dhataki 9 18.22 3.270

300 µg/µl Yashtimadh 9 1.11 3.333 - 10.930 0.000 HS

Dhataki 9 17.22 2.906


Dhataki 9 15.00 2.449


Dhataki 9 11.89 1.691



Page 129

S - significant; HS - Highly significant

Comparing the same concentration of aqueous extract of Yashtimadhu and Dhataki by unpaired

t- test it was observed that in all the concentration were found to be statistically highly

significant.



Page 130


Table No – 57: Colony Characterization and Microscopy of Pseudomonas

aeruginosa


Growth onMacCONKEY

Agar

CultureCharacters


Organismidentified


Growth seen

Size(in mm)

2 -3 mm

Gramnegative

Pseudomonasaeruginosa

Shape Circular

Surface Smooth

Elevation LowConvex

Opacity Opaque

Non- lactose fermentingcolonies



Page 131


extract of Yashtimadhu against Pseudomonas aeruginosa


concentrations of Yashtimadhu against Pseudomonas aeruginosa


Extract *ZOI in mm

against

Pseudomonas

aeruginosa

S

600 µg/µl

N=8

500 µg/µl

N= 8

400 µg/µl

N= 8

300 µg/µl

N= 8

200 µg/µl

N= 8

100 µg/µl

N= 8

F % F % F % F % F % F %

Aaq Aqueous

extr extract of

Ya Yashtimadhu

0 5 62.5 5 62.5 6 75 6 75 6 75 6 75

7 1 12.5 1 12.5

10 1 12.5 1 12.5 1 12.5 1 12.5 1 12.5 1 12.5

12 1 12.5 1 12.5

13 1 12.5

111 15 1 12.5

16 1 12.5

18 1 12.5



Page 132


200, 100 µg/µl) against Pseudomonas aeruginosa in 8 samples isolated by culture and

sensitivity has drawn the following data.

At 600 µg/µl Yashtimadhu showed maximum zone of inhibition was 16 mm in 1 (12.5%)

sample, 15 mm in 1 (12.5%) sample and minimum zone of inhibition 10 mm was observed in

1(12.5%) sample and there was no zone of inhibition observed in 5 (62.5%) samples.


13mm in 1(12.5%) sample and minimum zone of inhibition 10mm was observed in 1(12.5%)

sample and there was no zone of inhibition observed in 5 (62.5%) samples.


(12.5%) sample and there was no zone of inhibition observed in 6 (75%) samples.









Page 133


extract of Dhataki against Pseudomonas aeruginosa


concentrations of Dhataki against Pseudomonas aeruginosa

Extract *ZOI in mm

against

Pseudomonas

aeruginosa

S

600 µg/µl

N=8

500 µg/µl

N= 8

400 µg/µl

N= 8

300 µg/µl

N= 8

200 µg/µl

N= 8

100 µg/µl

N= 8

F % F % F % F % F % F %

Aq Aqueous

extr extract of

Ya Dhataki

10 1 12.5 3 37.5

12 2 25 2 25

13 1 12.5 1 12.5 1 12.5

15 1 12.5 4 50 3 37.5 1 12.5

16 2 25 4 50 1 12.5

17 3 37.5 1 12.5

18 1 12.5 1 12.5 1 12.5

20 2 25 3 37.5 3 37.5 1 12.5

21 1 12.5

23 1 12.5 1 12.5



Page 134



100 µg/µl) against Pseudomonas aeruginosa in 8 samples isolated by culture and sensitivity

has drawn the following data.

At 600 µg/µl Dhataki showed maximum zone of inhibition was 27 mm in 1 (12.5%) sample,

25 mm in 1 (12.5%) sample, 23 mm in 1 sample (12.5%), 21 mm in 1 (12.5%) sample, 20 mm

in 2(25%) samples and minimum zone of inhibition 16 mm was observed in 2 (25%) samples.


mm in 3 samples (37.5%), 18 mm in 1 (12.5%) sample and minimum zone of inhibition 17

mm was observed in 3 (37.5%) samples.


16 mm in 4 (50%) samples and minimum zone of inhibition 15 mm was observed in 1 (12.5%)

sample.


18 mm in 1 (12.5%) sample, 16 mm in 1 (12.5%) sample, 15 mm in 4 (50%) samples and

minimum zone of inhibition 13 mm was observed in 1 (12.5%) sample.


15 mm in 3 (37.5%) samples, 13 mm in 1 (12.5%) sample, 12 mm in 2 (25%) samples and

minimum zone of inhibition 10 mm was observed in 1 (12.5%) sample.

25 1 12.5

27 1 12.5



Page 135


mm in 1 (12.5%) sample, 13 mm in 1 (12.5%) sample, 12 mm in 2 (25%) samples and


Table No – 60: Comparing antibacterial action of aqueous extracts of Yashtimadhu

and Dhataki against Pseudomonas aeruginosa


mean difference between the zone of inhibitions of aqueous extracts of Yashtimadhu and






5.13 5.13 2.75 2.75 2.13 2.13


21.00 19.00 17.38 15.88 13.63 12.50




Page 136


Dhataki against Pseudomonas aeruginosa



Dhataki is having better antibacterial action against Pseudomonas aeruginosa than Dhataki.

For different concentrations of both Yashtimadhu and Dhataki, it is evident that increasing the

concentrations significant increase in zone of inhibition. Consecutively number of sensitive

zones increasing and number of resistant zones decreasing when increasing the concentrations.




2 1 5 2 1 5 1 1 6 0 2 6 0 2 6 0 2 6


8 0 0 8 0 0 8 0 0 7 1 0 4 4 0 2 6 0



Page 137

STATISTICS



unpaired t- test



Pseudo-

monas

Yashtimadhu 600 µg/µl 8 5.13 7.279 0.500 0.774 NS

500 µg/µl 8 5.13 7.396

400 µg/µl 8 2.75 5.120

300 µg/µl 8 2.75 5.120

200 µg/µl 8 2.13 4.016

100 µg/µl 8 2.13 4.016

Dhataki 600 µg/µl 8 21.00 3.928 11.679 0.000 HS

500 µg/µl 8 19.00 2.138

400 µg/µl 8 17.38 2.200

300 µg/µl 8 15.88 2.167



Page 138

200 µg/µl 8 13.63 2.264

100 µg/µl 8 12.50 2.828


NS - Not significant; HS - Highly significant


0.774 which is not statistically significant . Dhataki has P value 0.000 which is highly

significant. Futher on comparing the mean values, the mean value of Dhataki was greater than

the mean value of Yashtimadhu in all the concentrations.

Table No - 63: Statistical values of Unpaired t - test

Conc Group N Mean SD t test value P Remarks

Pseudo-

monas


Dhataki 8 21.00 3.928


Dhataki 8 19.00 2.138


Dhataki 8 17.38 2.200

300 µg/µl Yashtimadh 8 2.75 5.120 - 6.667 0.000 HS

Dhataki 8 15.88 2.167


Dhataki 8 13.63 2.264



Page 139




unpaired t- test it was observed that in all the concentration were found to be statistically

highly significant.


Dhataki 8 12.50 2.828

DISCUSSION


DISCUSSION

The present study “Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as

pratinidhi dravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference to

vranahara karma” was selected based on the Vranahara karma of two drugs, Aqueous

extract of Yashtimadhu (Glycyrrhiza glabra Linn.) and Aqueous extract of Dhataki

(Woodfordia fruticosa (L.) Kurz). Their antibacterial activity was evaluated by Culture

and Sensitivity Test. The observed results of drug Woodfordia fruticosa (L.) Kurz was

compared as a substitute with Glycyrrhiza glabra Linn and is discussed under following

heading.

1. Discussion on Drug

2. Discussion on Disease

3. Discussion on Pharmacognostic study

4. Discussion on In vitro study

Discussion on Drug

Glycyrrhiza glabra Linn a perennial under shrub belonging to Papilionaceae is one of the

most widely used herb. Glycyrrhiza glabra Linn has many therapeutic utility, it is used

for the treatment of fever, in upper respiratory tract ailment such as cough, sore throat,

bronchitis. It is also used as an antacid, laxative, in wound healing, to enhance

complexion of skin, etc. It is used as rasayana, antistress and anabolic agent.157 It also

has various other uses it has its utility in food, cosmetic, feed, fibre material, dyeing and

tobacco industries.158 As none of the liquorice – yielding species occurs in India.8 The

drug is imported into India from Asia minor, Iraq, Persia and other Central Asian

DISCUSSION


countries.8 The part used is root which is a destructive form of harvesting.1 To meet the

high demand the drug is over exploited and also adulterated with roots of Abrus

precatorius Linn and roots of Glycyrrhiza uralensis Fish (Manchurian liquorice) . Stem

pieces of Glycyrrhiza glabra Linn are also sold in place of root.9 Glycyrrhiza glabra Linn

comes under threatened/ depleted ( R. N. Chopra’s list).10 Glycyrrhiza glabra Linn is an

endangered medicinal plant and has been placed in red data book.11 The active of

Glycyrrhiza glabra Linn i.e Glycyrrhizine cannot be produced synthetically.12

Yashtimadhu can be seen as one of the ingredient mentioned in Tiktadhya ghrta,22

Patolyadi lepa,23 Aswagandhadi lepa,21 Jatyadi taila.24 These are mainly indicated in

vranaropana, vranashotha, vranashodhana. It is having madhura rasa, guru snigdha

guna, sheeta veerya, madhura vipaka and its dosha karma is vatapitta shamaka.159

Woodfordia fruticosa (L.) Kurz is described since Vedic literature. The drug was being

used more in Sandhana Kalpana. Woodfordia fruticosa (L.) Kurz is found throughout

India.13 The drug is used externally for its krimighna, kusthaghna, vranaropana. The

drug is indicated in Atisara, krimi, raktapitta, trushna, visarpa, visha. Its flowers being

the used part of the plant is mentioned in various preparation used in vrana.18- 24Dhataki

can be seen as one of the ingredient in Dhatakyadi churna,18 Kumbhikadhya taila,19

Panchavalkaladhya churna,20 Ashwagandhadi lepa.21 These are mainly indicated in

vranashotha, vranaropana, vranashodhana. It is having kashaya rasa, laghu ruksha guna,

sheetavirya, katu vipaka and its dosha karma is kapha pitta shamaka.160

Ayurvedic literature mentions Dhatakipushpa (Woodfordia fruticosa (L.) Kurz) as the

substitute of Yashtimadhumoola (Glycyrrhiza glabra Linn).3- 5 Both Yashtimadhu and

DISCUSSION


Dhataki having pitta shamaka property, possesses Vranahara, Vishahara, Trishnahara

karmas similar to each other.14- 17 Their action on Vranahara karma can be substantiated

based on its properties.161 As Yashtimadhu is having madhura rasa which is vata pitta

shamana. By pitta shamana it helps to reduce daha and shotha . Its sheeta virya will help

in stambhana of rakta. Due to its snigdha guna there will be proper ropana. Snigdha

guna and madhura vipaka help to reduce pain caused by vata. Tikta rasa helps in lekhana,

kleda soshaka and does shodhana. Dhataki - kashaya rasa has ruksha guna and brings

about kleda shoshana, ropana and lekhana. Laghu guna is helpful in lekhana. Katu

vipaka cause shoshana of kapha, kleda due to its ruksha and laghu guna. Sheeta veerya

does stambhana and reduce daha. This will give shodhana of dushta vrana

Ushna guna due to its teekshna guna is krimighna.

Discussion on Disease

Acharya Sushruta has described Vrana and its management in detail. Broadly Vrana is

classified into two types Dushta Vrana (chronic ulcer / non healing ulcer) and Shuddha

Vrana. A wound which refuses to heal or heals very slowly in spite of best efforts is

known as Dushta Vrana. Such Vrana emits foul smell, continuously putrified pus flows

along with blood, presents with cavity, will be long standing etc, pronounced Vrana

Lakshana and characters opposite to Shuddha Vrana is termed as Dushta Vrana. Hence it

may be inferred that definite knowledge of nonhealing ulcer was prevailing during

Samhita period. Dushta Vrana (non healing diabetic ulcer) is most common type of

infected ulcer in human life and gets infected with bacteria. The treatment for non

DISCUSSION


healing ulcer is antimicrobial therapy associated with side effects or limitations. The

infected person becomes drug resistant if the drug is improperly administered.

Antimicrobial resistance threatens the effective prevention and treatment of an ever

increasing range of infections caused by bacteria, viruses and fungi. It is an increasingly

serious threat to humanity. There are high proportions of antibiotic resistance in bacteria

that causes common infections. So that standard treatments become ineffective and

infections persist, increasing the risk of spreading to others. Variety of chemical

substances have been evaluated and patented as wound cleansing and healing agents,

their inability to become successful drugs is due to the fact that these are able to act only

at a particular step of the cleansing and healing cascade. It is therefore, necessary that

these agents of optimum biological activity have to be based on a specific mechanism of

the healing process. It is likely that more effective wound debridement and healing agents

can be developed from natural products. Ayurvedic classics has advocated many Kashaya,

Taila, Ghrita Lepa, etc. medicament for encouraging wound debridement and healing

agents. The effect of these is due to action of the constituent drug active principles as a

whole. These act on the microorganisms. In Ayurveda various drugs are mentioned for

Dushta Vrana which is also ascribed with Krimighna action, but indication of specific

drug in specific causative microorganism and stage of disease is missing and very few

studies have been accomplished. Therefore it is the need of the hour to use diagnostic

tools like culture and sensitivity and identify causative micro-organism, its characteristics

and other attributes, further culture these organisms in vitro and study the sensitivity.

Pharmacognostical study

DISCUSSION


Physicochemical evaluation.

Foreign matter

Foreign matter in Glycyrrhiza glabra Linn. was absent and in Woodfordia fruticosa

(L.) Kurz. is 0.17% which is as per the standard of Quality standard of Indian Medicinal

Plants.

Loss on Drying

This is the method to determine the moisture content of a drug. It aids to prevent the

decomposition of the drugs either due to chemical change or microbial contamination.

From the results obtained for loss of drying, it was found to be 12% in Glycyrrhiza

glabra Linn. which is as per standard of API and 13.1% in Woodfordia fruticosa (L.)

Kurz .

Determination of Ash value

Ash value is the criterion to judge authenticity and purity of crude drugs. The residue

remaining after incineration is the ash content of the drug. These could be inorganic salts

such as carbonates, sulphates, phosphates, silicates etc. naturally occurring in the drug or

adhered to it or deliberately added to it in order to adulterate the drug. Total ash is to

measure the total amount of plant material remaining after ignition of the drug. Acid

insoluble ash is the residue obtained after boiling the total ash either with dilute

hydrochloric acid or water which measures the amount of sand and silica matter present

in the drug.

DISCUSSION


The study showed that Total ash value for Glycyrrhiza glabra Linn. as 7.5% which is as

per standard of API and 6% was the observed ash value in Woodfordia fruticosa (L.)

Kurz which is as per standard of Quality standard of Indian Medicinal Plants. Acid

insoluble ash in Glycyrrhiza glabra Linn. is 1% which is as per standard of API and in

Woodfordia fruticosa (L.) Kurz is 8% which is more when compared to Quality standard

of Indian Medicinal Plants, this is due to the presence of other than the used part of the

drug while collecting.

Determination of extractive value

Extractive value measures the nature of the chemical constituents present in a crude drug.

It is essential for the estimation of specific chemical constituents soluble in that particular

solvent used for extraction.

Alcoholic extract value in in Glycyrrhiza glabra Linn. is 10% which is as per standard

of API and in Woodfordia fruticosa (L.) Kurz. is 8% which is as per standard of Quality

standard of Indian Medicinal Plants.

Water extractive value in Glycyrrhiza glabra Linn is 31% which is as per standard of

API and in Woodfordia fruticosa (L.) Kurz is 41% which is as per standard of Quality

standard of Indian Medicinal Plants.

DISCUSSION


Preliminary phytochemical analysis:

The study showed Carbohydrate, Tannin, Flavanoids, Saponins, Phenols, Carboxylic acid,

Quinone are common chemical constituents in both Glycyrrhiza glabra Linn. and

Woodfordia fruticosa (L.) Kurz.

Alkaloids , Steroids, Triterpenoid, Resins are present only in Glycyrrhiza glabra Linn.

Coumarins, Amino acids are absent in both Glycyrrhiza glabra Linn. and Woodfordia

fruticosa (L.) Kurz.

According to previous studies- Tannins, Flavonoids, Saponins, Phenols, Carbohydrates,

Carboxylic acids, Quinone showed antimicrobial properties.162- 170

Saponins have been reported to possess a wide range of biological activities. The mode of

action of antibacterial effects of saponins seems to involve membranolytic properties,

rather than simply altering the surface tension of the extracellular medium, thus being

influenced by microbial population density (Killeen et al., 1998).171 The action

mechanisms of saponins may lie in damage to the membrane and leakage of cellular

materials, ultimately leading to cell death (Mshvildadze et al., 2000).172 Flavonoids are

phenolic structures containing one carbonyl group. Their activity is probably due to their

ability to complex with extracellular and soluble proteins and to complex with bacterial

cell walls (Cowan, 1999).173 More lipophilic flavonoids may also disrupt microbial

membranes (Tsuchiya et al., 1996).174 Alkaloids isolated from plant are commonly found

to have antimicrobial properties (Omulokoli et al., 1997).175 Berberine and harmane are

important representatives of the alkaloid group. Their mechanism of action is attributed

to their ability to intercalate with DNA of bacteria (Phillipson and O′Neill, 1987).176 The

DISCUSSION


astringent character of tannins may induce complexation with enzymes or substrates.

Many microbial enzymes were found to be inhibited when raw culture filtrates or purified

enzymes were mixed with tannins.177 Quinones are known to complex irreversibly with

nucleophilic amino acids in proteins , often leading to inactivation of the protein and loss

of function. For that reason, the potential range of quinone antimicrobial effects is great.

Probable targets in the microbial cell are surface-exposed adhesins, cell wall polypeptides,

and membrane-bound enzymes. Quinones may also render substrates unavailable to the

microorganism.173

HPTLC

The bands run through TLC with numerous Rf values represents the individual

phytoconstituents. In the result common Rf value of the both plants is detected.

In Dhataki using Solvent system – Toluene: Chloroform: Ethyl Acetate: Formic acid (2:

6: 6 : 2) and Yashtimadhu using Solvent system – n- butanol: Water: Glacial acetic acid

(7: 2: 1) , the common active component was detected in Rf value 0. 05 which occupied

2.17% (254 nm) and 1.80% (620 nm) area in Glycyrrhiza glabra Linn. and 54.08% (254

nm) area in Woodfordia fruticosa (L.) kurz. Rf value 0.24 which occupied 53.01% (620

nm) area in Glycyrrhiza glabra Linn. and 8.06% (366 nm) area in Woodfordia fruticosa

(L.) Kurz. Rf value 0.42 which occupied 7.68% (620 nm) in Glycyrrhiza glabra Linn.

and 3.36% (620 nm ) area in Woodfordia fruticosa (L.) Kurz. Rf value 0.72 which

occupied 35.85% (366 nm) in Glycyrrhiza glabra Linn. and 0.71% (366 nm) area in

Woodfordia fruticosa (L.) Kurz .

DISCUSSION


In Dhataki using Solvent system – Toluene: Ethyl Acetate: Methanol: Formic acid

(3: 3: 8: 2) and Yashtimadhu using Solvent system – n- butanol: Water: Glacial acetic

acid (7: 2: 1) , the common active component was detected in Rf value 0.05 which

occupied 2.17% (254 nm) area and 1.80% (620 nm) area in Glycyrrhiza glabra Linn.

and 74.57% (254 nm) area and 27.42% (620 nm) area in Woodfordia fruticosa (L.) Kurz.

Rf value 0.20 which occupied 4.87% (366 nm) area in Glycyrrhiza glabra Linn. and

9.13% (366 nm) area in Woodfordia fruticosa (L.) Kurz.

The common and nearer value is required to bring same or nearer therapeutic effect. The

results from HPTLC finger prints, scanned at various wavelengths revealed that there are

many compounds in both drugs i.e. Glycyrrhiza glabra Linn & Woodfordia fruticosa (L.)

Kurz. From those chromatograms, it has been found that alcoholic extracts of either of

the drugs contain multiple spots and peaks representing multiple components responsible

for specific therapeutic effect in the body. Major therapeutic effect is brought by the Rf

value which are in Higher concentration.178

These results provide a very important clue for further isolation of pure compounds

which are shown in higher concentration in chromatography to elicit the exact bio

chemical compound involve in performing the respective activities.

DISCUSSION


Experimental Study – In vitro

In the present study, antibacterial activity of aqueous extracts of Yashtimadhu and

Dhataki was analyzed against Staphylococcus aureus, Escheriochia coli, Acinetobacter

and Pseudomonas aeruginosa. Among the 60 pus sample , Staphylococcus aureus was

the most common organisms followed by Escherichia coli. The assay was done at 6

different concentrations of aqueous extracts of Yashtimadhu and Dhataki to understand

their effective activity. Here anti - bacterial study is done by Agar well diffusion method.


In order to evaluate efficacy and conclude efficacy of Yashtimadhu (Glycyrrhiza glabra Linn.)

and Dhataki (Woodfordia fruticosa (L.) Kurz.), it was considered that for Yashtimadhu the zone

of inhibition from 20- 16 mm is taken as sensitive zone, 14-10 mm is taken as moderately

sensitive, 8- 4 mm mm is taken as resistant. For Dhataki the zone of inhibition from 26- 22mm

is taken as sensitive, 20- 16 mm is taken as moderately sensitive, 14- 10 mm as resistant.

600 µg/µl was the highest concentration of aqueous extract of Yashtimadhu and Dhataki taken

respectively, which showed significant zone of inhibition against Staphylococcus aureus. Out of

25 samples, in aqueous extract of 600 µg/µl of Yashtimadhu, 14 samples are sensitive, 3 sample

is moderately sensitive and 8 samples are resistant. The mean value of zone of inhibition is

11.56 mm. Whereas in aqueous extract of 600 µg/µl of Dhataki, 14 samples are sensitive, 10

samples are moderately sensitive and 1 samples is resistant. The mean value of zone of

inhibition is 21.04 mm. Thus at 600 µg/µl concentration, aqueous extract of Dhataki is better or

more sensitive than Yashtimadhu against Staphylococcus aureus.

DISCUSSION


In 500 µg/µl concentration of aqueous extract of Yashtimadhu and Dhataki taken respectively,

which showed significant zone of inhibition against Staphylococcus aureus. Out of 25 samples,

in aqueous extract of 500 µg/µl of Yashtimadhu, 12 samples are sensitive, 2 sample is

moderately sensitive and 11 samples are resistant. The mean value of zone of inhibition is 9.76

mm. Whereas in aqueous extract of 500 µg/µl of Dhataki, 11 samples are sensitive, 13 samples

are moderately sensitive and 1 samples is resistant. The mean value of zone of inhibition is

20.40 mm. Thus at 500 µg/µl concentration, aqueous extract of Dhataki is better or more

sensitive than Yashtimadhu against Staphylococcus aureus.

In 400µg/µl concentration of aqueous extract of Yashtimadhu and Dhataki taken respectively,


in aqueous extract of 400 µg/µl of Yashtimadhu, 9 samples are sensitive, 6 sample is moderately

sensitive and 10 samples are resistant. The mean value of zone of inhibition is 8.96 mm.

Whereas in aqueous extract of 400 µg/µl of Dhataki, 8 samples are sensitive, 16 samples are

moderately sensitive and 1 samples is resistant. The mean value of zone of inhibition is 19.20

mm. Thus at 400 µg/µl concentration, aqueous extract of Dhataki is better or more sensitive

than Yashtimadhu against Staphylococcus aureus.






moderately sensitive and 8 samples is resistant. The mean value of zone of inhibition is

DISCUSSION


18.32mm. Thus at 300 µg/µl concentration, aqueous extract of Dhataki is better or more


















Hence from all these results it is clear that Dhataki has better antibacterial action than

Yashtimadhu against Staphylococcus aureus isolated from the pus samples of the patients

suffering from Dushta vrana

DISCUSSION


The aqueous extracts of both Yashtimadhu and Dhataki had shown significant antibacterial

action against Staphylococcus aureus. Here in the study, for aqueous extract of Yashtimadhu the

maximum zone of inhibition seen was 28 mm which was shown by 600 µg/µl concentration

and minimum zone of inhibition was 7 mm which was recorded at 100µg/µl. For aqueous

extract of Dhataki, the maximum zone of inhibition seen was 30 mm at 400 µg/µl and 200 µg/µl

concentrations and minimum zone of inhibition seen was 10 mm recorded at 200 µg/ µl and 100

µg/µl concentrations.

For different concentrations of both Yashtimadhu and Dhataki, it was evident that increasing the

concentrations there was significant increase in zone of inhibition i.e Staphylococcus aureus

inhibited their growth with the least dilution or with higher concentration of the plant extracts.

This was due to the small proportion of water in the mixture solvent which might have increase

the quality or the quantity of the phytochemical bioactive components of the plant extracts.173, 179

Statistical Analysis

Statistical analysis of the data was performed using SPSS 23.0 (IBM corp) using One-Way

ANOVA test followed by Unpaired t - test. P = 0.01- 0.001 is considered as statistically highly

significant, P = 0.01-0.05 is considered as statistically significant and P > 0.05 is considered as

not significant. On comparing between Yashtimadhu and Dhataki by One way Anova,

Yashtimadhu has P value 0.114 which is not statistically significant. Dhataki has P value 0.000

which is highly significant. The probable reason for Yashtimadhu showing non-significant

(p=0.114) result over Dhataki (highly significant at p=0.000) may be due to the variance in the

percentage of sensitivity at various concentrations. Further on comparing the mean values,

mean value of Dhataki was greater than mean value of Yashtimadhu in all the different

DISCUSSION


concentrations. Further on Comparing the same concentration of aqueous extract of

Yashtimadhu and Dhataki by unpaired t - test it was observed that in all the concentration were

found to be statistically highly significant. It was found that at similar concentration both

Yastimadhu and Dhataki showed highly significant results, this may be due to the presence of

definite antibacterial activity in terms of its percentage sensitivity. Hence at similar

concentration we can derive that both Dhataki (Woodfordia fruticosa (L.) Kurz.) and

Yastimadhu (Glycyrrhiza glabra Linn.) possesses antibacterial property individually.

Escherichia coli


and Dhataki (Woodfordia fruticosa (L.) Kurz.), it was considered for Yashtimadhu the zone of

inhibition from 14- 12 mm is taken as sensitive zone, 10-8 mm is taken as moderately sensitive,

6- 4 mm is taken as resistant. For Dhataki the zone of inhibition from 24- 20mm is taken as

sensitive, 18- 14 mm is taken as moderately sensitive, 12- 8 mm as resistant.


respectively, which showed significant zone of inhibition against Escherichia coli. Out of 18

samples, in aqueous extract of 600 µg/µl of Yashtimadhu, 1 samples are sensitive, No samples

are moderately sensitive and 17 samples are resistant. The mean value of zone of inhibition is

0.67 mm. Whereas in aqueous extract of 600 µg/µl of Dhataki, 12 samples are sensitive, 5

samples are moderately sensitive and 1 samples is resistant. The mean value of zone of

inhibition is 20.33 mm. Thus at 600 µg/µl concentration, aqueous extract of Dhataki is better or

more sensitive than Yashtimadhu against Escherichia coli.

DISCUSSION



which showed significant zone of inhibition against Escherichia coli. Out of 18 samples, in

aqueous extract of 500 µg/µl of Yashtimadhu, No samples are sensitive, 1 sample is moderately



moderately sensitive and 1 samples is resistant. The mean value of zone of inhibition is 19.44


than Yashtimadhu against Escherichia coli.



aqueous extract of 400 µg/µl of Yashtimadhu, 2 samples are sensitive, 1 sample is moderately



moderately sensitive and No samples are resistant. The mean value of zone of inhibition is 19.39





aqueous extract of 300 µg/µl of Yashtimadhu, 1 samples are sensitive, No samples are


mm. Whereas in aqueous extract of 300 µg/µl of Dhataki, 7 sample are sensitive, 11 samples are

moderately sensitive and No samples are resistant. The mean value of zone of inhibition is 18

DISCUSSION






aqueous extract of 200 µg/µl of Yashtimadhu, 2 samples are sensitive, 2 samples are moderately


Whereas in aqueous extract of 200 µg/µl of Dhataki, 6 sample are sensitive, 9 samples are






aqueous extract of 100 µg/µl of Yashtimadhu, 2 samples are sensitive, 2 samples are moderately


Whereas in aqueous extract of 100 µg/µl of Dhataki, 4 sample are sensitive, 5 samples are





Yashtimadhu against Escherichia coli isolated from the pus samples of the patients suffering

from Dushta vrana

DISCUSSION



action against Escherichia coli. Here in the study, for aqueous extract of Yashtimadhu the


and minimum zone of inhibition was 7 mm which was recorded at 200µg/µl. For aqueous

extract of Dhataki, the maximum zone of inhibition seen was 29 mm at 600 µg/µl, 500 µg/µl

and 300 µg/µl concentrations and minimum zone of inhibition seen was 10 mm recorded at

200 µg/ µl and 100 µg/µl concentrations.

For different concentrations of Yashtimadhu it was shown that increase in the

concentrations there was a decrease in zone of inhibition i.e Escherichia coli inhibited its

growth with higher serial dilution or with lower concentration of the plant extracts. It was

seen that there is not much difference in zone of inhibition between the different

concentrations of Yashtimadhu , this can be due to the fact that within the same species of

Escherichia coli there maybe presence of different strains which might vary the virulence

level.180 For different concentrations of Dhataki it was shown that increase in the

concentrations there was increase in zone of inhibition, this might be due to the presence

of more bioactive component of the plant extracts at higher concentration.173, 179






Yashtimadhu has P value 0.345 which is not statistically significant . Dhataki has P value 0.001

DISCUSSION



(p=0.345) result over Dhataki (highly significant at p=0.001) may be due to the variance in the




Yashtimadhu and Dhataki by Unpaired t - test it was observed that in all the concentration

were found to be statistically highly significant, this may be due to the presence of definite

antibacterial activity in terms of its percentage sensitivity. Hence at similar concentration we

can derive that both Dhataki (Woodfordia fruticosa (L.) Kurz.) and Yastimadhu (Glycyrrhiza

glabra Linn.) possesses antibacterial property individually.

Acinetobacter




6- 4 mm is taken as resistant. For Dhataki the zone of inhibition from 24- 20mm is taken as



respectively, which showed significant zone of inhibition against Acinetobacter. Out of 9

samples, in aqueous extract of 600 µg/µl of Yashtimadhu, 3 samples are sensitive, 1 sample is



DISCUSSION


are moderately sensitive and No samples are resistant. The mean value of zone of inhibition is


sensitive than Yashtimadhu against Acinetobacter.

In 500 µg/µl concentration of aqueous extract of Yashtimadhu and Dhataki taken

respectively, which showed significant zone of inhibition against Acinetobacter. Out of 9

samples, in aqueous extract of 500 µg/µl of Yashtimadhu, 3 samples are sensitive, 1 sample is



are moderately sensitive and No samples are resistant. The mean value of zone of inhibition

is 20.33 mm. Thus at 500 µg/µl concentration, aqueous extract of Dhataki is better or more



which showed significant zone of inhibition against Acinetobacter. Out of 9 samples, in

aqueous extract of 400 µg/µl of Yashtimadhu, 1 sample is sensitive, 2 sample are moderately



moderately sensitive and No samples are resistant. The mean value of zone of inhibition is





aqueous extract of 300 µg/µl of Yashtimadhu, No samples are sensitive, 1 sample is

DISCUSSION



mm. Whereas in aqueous extract of 300 µg/µl of Dhataki, 2 samples are sensitive, 7 sample

are moderately sensitive and No samples are resistant. The mean value of zone of inhibition

is 17.22 mm. Thus at 300 µg/µl concentration, aqueous extract of Dhataki is better or more




aqueous extract of 200 µg/µl of Yashtimadhu, No samples are sensitive, 1 sample is


mm. Whereas in aqueous extract of 200 µg/µl of Dhataki, 1 sample is sensitive, 7 samples are

moderately sensitive and 1 sample is resistant. The mean value of zone of inhibition is 15 mm.

Thus at 200 µg/µl concentration, aqueous extract of Dhataki is better or more sensitive than

Yashtimadhu against Acinetobacter.



aqueous extract of 100 µg/µl of Yashtimadhu, No samples are sensitive, No samples are

moderately sensitive and 9 samples are resistant. The mean value of zone of inhibition is 0

mm. Whereas in aqueous extract of 100 µg/µl of Dhataki, No samples are sensitive, 2

samples are moderately sensitive and 7 samples are resistant. The mean value of zone of

inhibition is 11.89 mm. Thus at 100 µg/µl concentration, aqueous extract of Dhataki is better

or more sensitive than Yashtimadhu against Acinetobacter.

DISCUSSION



Yashtimadhu against Acinetobacter isolated from the pus samples of the patients suffering

from Dushta vrana


action against Acinetobacter. Here in the study, for aqueous extract of Yashtimadhu the


and minimum zone of inhibition was 10 mm which was recorded at 600µg/µl, 500 µg/ µl,

400 µg/ µl, 300 µg/ µl, 200 µg/ µl concentrations. For aqueous extract of Dhataki, the

maximum zone of inhibition seen was 25 mm at 600 µg/µl and 500 µg/µl concentrations and

minimum zone of inhibition seen was 10 mm recorded at 100 µg/µl concentration.

For different concentrations of both Yashtimadhu (Glycyrrhiza glabra Linn.) and

Dhataki(Woodfordia fruticosa (L.) Kurz.), it was evident that increasing the

concentrations there was significant increase in zone of inhibition i.e Acinetobacter

inhibited their growth with the least dilution or with higher concentration of the plant

extracts. This was due to the small proportion of water in the mixture solvent which

might have increase the quality or the quantity of the phytochemical bioactive

components of the plant extracts.173, 179






DISCUSSION


Yashtimadhu has P value 0.055 which is statistically significant . Dhataki has P value 0.000

which is highly significant. Further on comparing the mean values, mean value of Dhataki was

greater than mean value of Yashtimadhu in all the different concentrations. Further on


Unpaired t - test it was observed that in all the concentration were found to be statistically

highly significant. Hence we can derive that both Dhataki (Woodfordia fruticosa (L.) Kurz.)

and Yastimadhu (Glycyrrhiza glabra Linn.) possesses antibacterial property.





6- 4 mm is taken as resistant. For Dhataki the zone of inhibition from 20- 16 mm is taken as



respectively, which showed significant zone of inhibition against Pseudomonas aeruginosa. Out

of 8 samples, in aqueous extract of 600 µg/µl of Yashtimadhu, 2 samples are sensitive, 1 sample

is moderately sensitive and 5 samples are resistant. The mean value of zone of inhibition is 5.13

mm. Whereas in aqueous extract of 600 µg/µl of Dhataki, 8 samples are sensitive, No samples

are moderately sensitive and No samples are resistant. The mean value of zone of inhibition is

21 mm. Thus at 600 µg/µl concentration, aqueous extract of Dhataki is better or more sensitive

than Yashtimadhu against Pseudomonas aeruginosa.

DISCUSSION



which showed significant zone of inhibition against Pseudomonas aeruginosa. Out of 8 samples,


sensitive and 5 samples are resistant. The mean value of zone of inhibition is 5.13 mm. Whereas

in aqueous extract of 500 µg/µl of Dhataki, 8 samples are sensitive, No samples are moderately

sensitive and No samples are resistant. The mean value of zone of inhibition is 19 mm. Thus at

500 µg/µl concentration, aqueous extract of Dhataki is better or more sensitive than

Yashtimadhu against Pseudomonas aeruginosa.



in aqueous extract of 400 µg/µl of Yashtimadhu, 1 samples is sensitive, 1 sample is moderately

sensitive and 6 samples are resistant. The mean value of zone of inhibition is 2.75 mm. Whereas

in aqueous extract of 400 µg/µl of Dhataki, 8 samples are sensitive, No samples are moderately

sensitive and No samples are resistant. The mean value of zone of inhibition is 17.38 mm. Thus

at 400 µg/µl concentration, aqueous extract of Dhataki is better or more sensitive than

Yashtimadhu against Pseudomonas aeruginosa.



in aqueous extract of 300 µg/µl of Yashtimadhu, No samples are sensitive, 2 samples are


mm. Whereas in aqueous extract of 300 µg/µl of Dhataki, 7 samples are sensitive, 1 samples is


DISCUSSION








mm. Whereas in aqueous extract of 200 µg/µl of Dhataki, 4 samples are sensitive, 4 samples are








mm. Whereas in aqueous extract of 100 µg/µl of Dhataki, 2 samples are sensitive, 6 samples are





Yashtimadhu against Pseudomonas aeruginosa isolated from the pus samples of the patients

suffering from Dushta vrana

DISCUSSION


The aqueous extracts of both Yashtimadhu and Dhataki had shown significant

antibacterial action against Pseudomonas aeruginosa. Here in the study, for aqueous

extract of Yashtimadhu the maximum zone of inhibition seen was 18 mm which was

shown by 500 µg/µl concentration and minimum zone of inhibition was 7 mm which

was recorded at 200µg/µl and 100 µg/ µl concentrations. For aqueous extract of Dhataki,

the maximum zone of inhibition seen was 27 mm at 600 µg/µl concentration and

minimum zone of inhibition seen was 10 mm recorded at 200 µg/µl and 100 µg/µl

concentrations.

For different concentrations of both Yashtimadhu and Dhataki, it was evident that

increasing the concentrations there was significant increase in zone of inhibition i.e

Pseudomonas aeruginosa inhibited their growth with the least dilution or with higher

concentration of the plant extracts. This was due to the small proportion of water in the

mixture solvent which might have increase the quality or the quantity of the

phytochemical bioactive components of the plant extracts.173, 179






Yashtimadhu has P value 0.774 which is not statistically significant . Dhataki has P value 0.000


(P=0.774) result over Dhataki (highly significant at P=0.000) may be due to the variance in the

DISCUSSION





Yashtimadhu and Dhataki by unpaired t - test it was observed that in all the concentration

were found to be statistically highly significant. It was found that at similar concentration both

Yastimadhu and Dhataki showed highly significant results, this may be due to the presence of

definite antibacterial activity in terms of its percentage sensitivity. Hence at similar

concentration we can derive that both Dhataki (Woodfordia fruticosa (L.) Kurz.) and

Yashtimadhu (Glycyrrhiza glabra Linn.) possesses antibacterial property individually.

CONCLUSION


Page 166

CONCLUSION Yashtimadhu (Glycyrrhiza glabra Linn.) is one of the most widely used herb. The

plant is over exploited and its used part is root which is a destructive form of

harvesting. Ayurvedic literature mentions Dhatakipushpa (Woodfordia fruticosa L.

Kurz.) as the substitute of Yashtimadhumoola (Glycyrrhiza glabra Linn.).

The pharmacognostical studies have confirmed the authenticity and purity of

both the Yashtimadhu (Glycyrrhiza glabra Linn.) and Dhataki (Woodfordia

fruticosa L. Kurz.).

Both the drugs have few commom phytochemicals like carbohydrates, tannins,

flavanoids, saponins, phenol, carboxylic acid, quinone

According to previous studies- Tannins, Flavonoids, Saponins, Phenols,

Carbohydrates, Carboxylic acids, Quinone showed antimicrobial properties

In Dhataki (Woodfordia fruticosa L. Kurz.) using Solvent system – Toluene:

Chloroform: Ethyl Acetate: Formic acid (2: 6: 6 : 2) and Yashtimadhu

(Glycyrrhiza glabra Linn.) using Solvent system – n- butanol: Water: Glacial

acetic acid (7: 2: 1) the common active component was detected in Rf value 0.05,

0.24, 0.42, 0.72

In Dhataki (Woodfordia fruticosa L. Kurz.) using Solvent system – Toluene:

Ethyl Acetate: Methanol: Formic acid (3: 3: 8: 2) and Yashtimadhu (Glycyrrhiza

glabra Linn.)using Solvent system – n- butanol: Water: Glacial acetic acid (7: 2: 1)

the common active component was detected in Rf value 0.05, 0.20

CONCLUSION


Page 167

Staphylococcus aureus was the most common organisms found in wound pus

samples followed by Escherichia coli.

On comparisons the effect of aqueous extract of Dhataki (Woodfordia fruticosa L.

Kurz.) was found to be better when compared to aqueous extract of Yashtimadhu

(Glycyrrhiza glabra Linn.) based on statistical analysis

All the tested bacteria inhibited their growth with the least dilution, which means

high concentrations of the plant extract. This was due to the small proportion of

water in the mixture solvent which might have increase the quality or the quantity

of the phytochemical bioactive components of the plant extracts. Whereas in

Escherichia coli it was shown that for different concentrations of Yashtimadhu

(Glycyrrhiza glabra Linn.) inhibited its growth with higher serial dilution or with

lower concentration of the plant extract. This can be due to the fact that within the

same species of Escherichia coli there maybe presence of different strains which

might vary the virulence level .

The study shows that at similar concentration both drugs are having antibacterial

activity individually , this may be due to the presence of definite antibacterial

activity in terms of its percentage sensitivity.

Hence, hypothesis Dhataki (Woodfordia fruticosa (L.) Kurz.) can be used as

substitute for Yashtimadhu (Glycyrrhiza glabra Linn.) in Vranahara karma

(Antibacterial activity) in this study is justified.

CONCLUSION


Page 168

Limitation of Study:

Study was conducted on the detected few organisms from the few cases of

dushtavrana

Risks of contamination

Scope for further study:

Similar study can be conducted of specific organisms in big sample size

Clinical study

SUMMARY


SUMMARYThe present work entitled “Antibacterial activity of Dhataki (Woodfordia fruticosa (L.)

Kurz as pratinidhi dravya for Yashtimadhu (Glycyrrhiza glabra Linn) with reference to

vranahara karma” has been carried out. This study is divided into following sections -

introduction, review of literature, methodology, observations & results, discussion and

conclusion.

Introduction:

Aim, objectives, need and relevance of present study are elaborated in introduction.

Review of Literature:

Yashtimadhu (Glycyrrhiza glabra Linn.) and Dhataki (Woodfordia fruticosa (L.) Kurz.)

drugs were reviewed from samhithas, nighantus and recent text books on Dravyaguna

vignana, available pharmacopoeias. An updated research article search was added up to

the review of the relevant topics of the study. Disease and its management was review

from brihatrayee and laghutrayee followed by understanding the disease from

contemporary scientific view. Antibacterial activity was reviewed regarding the

morphology of bacteria, culture media , antibacterial mode of action .

Methodology

Methodology is divided into pharmacognostic, and experimental part. Pharmacognostic

study deals with the collection of drugs, authentication of drugs and preparation of

Extracts. Detailed study of Macroscopic, Microscopic, physicochemical, preliminary

SUMMARY


phytochemical study and HPTLC with densitometric scan of both drugs are also

documented.

At last, the experimental part includes the materials and methods of the study, procedure

implemented is explained.

Observations and results

This includes observation and results of the pharmacognostical study- its macroscopic,

microscopic, physicochemical analysis, phytochemical study, HPTLC with densitometric

scan of both drugs. The relevant readings of selected experimental study are represented

in the form of tables.

Discussion:

Based on review of literature, the observation and results of pharmacognostical study

and experimental study has been analyzed and discussed. The relevant logical reasoning

with scientific background is attempted in the discussion.

Conclusion:

This is a conclusive interpretation made on each study i.e. pharmacognostic and

experimental study from the outcome of the obtained data. It also includes the limitations

and further scope of the study.

REFERENCES

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz.) as pratinidhidravya for Yashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranaharakarma”

Page 171

REFERENCES

1. Sharma.S, Thokchom.R. A review on endangered medicinal plant of India

and their conservation. Journal of Crop and Weed. 2014; Vol 10(2) : p.205- 18

2. Ved DK, Goraya GS.Demand and supply of medicinal plants in India. New

Delhi: National Medicinal Plant Board; 2007

3. Misra Siddhi Nandan, editor. Bhaisajya Ratnavali of Kaviraj Govind Sen;

Abhavaprakarana: Chapter 4, Verse 29. Varanasi: Chaukhambha Surbharati

Prakashan, 2016; p. 72.

4. Shastri Brahmashankar Bhisagratna, editor. Yogaratnakara by Vaidya

Laksmipati Shastri, Purvaardhagata; Abhavavarga, Verse 13. Varanasi:

Chaukhambha Prakashan, 2015; p. 171

5. Misra Bhavamasankara, Vaisya Rupalalaji, editor, 9th. Bhavaprakasha of Sri

Bhavamisra Prathamabhaga; Verse 148. Varanasi: Chaukhambha Sanskrit

Sansthan, 1999; p. 182

6. BadkhaneYogesh,Yadav A.S, Bajaj. A, Sharma. AjitK,Raghuwanshi D.K.

Glycyrrhizaglabra L. A miracle medicinal herb . Indo American Journal of

Pharmaceutical Research. 2014; 4(12): p. 5808- 16

7. Dhar Uppendra, Rawa lRanbeer S, Upreti Jyoti .Setting priorities for

conservation of Medicinal plants a case study in the Indian Himalaya.

Biological Conservation . 2000; 95: p. 57- 65

8. Anonymous 1999. The wealth of India: a dictionary of Indian raw materials

and industrial products. Vol IV : F-G. CSIR, New Delhi. P. 151

REFERENCES


Page 172

9. SharmaP.C, Yelne M.B, DennisT.J.Database on Medicinal Plants Used in

Ayurveda Vol3. 2nd . New Delhi: Central Counsil for Research in Ayurveda &

Siddha; 2005: p.561- 6

10. Chaudhuri. A.B. Endangered Medicinal Plants. New Delhi: Daya Publishing

House; 2014: p. 111

11. Srivastava. M , et.al. In vitro conservation of Glycyrrhiza glabra by slow

growth culture. International Journal of Bio- Technology & Research. (IJBTR)

2013; Vol 3(1): p. 49- 58

12. KR. Akshay, N. Sudharanal, et.al. Biodiversity and strategies for conservation

of rare, endangered and threatened medicinal plants. Journal of

Pharmacognosy and Phytochemistry. 2014; Vol 2(3) : p. 12- 20

13. Pandey Gyanendra. Dravyaguna Vijnana Vol 1, 2nd . Varanasi: Krishnadas

Academy, 2002; p. 587







16. Sankhyadhar Satish Chandra, editor, 1st. Raj Nighantu of Sri Narhari Pandit,

Pippalyadivarga; Verse 144, 145. Varanasi: Chaukhambha Orientalia, 2012; p.

242

REFERENCES


Page 173

17. Sankhyadhar Satish Chandra, editor, 1st. Raj Nighantu of Sri Narhari Pandit,

Pippalyadivarga; Verse 213, 214. Varanasi: Chaukhambha Orientalia, 2012; p.

269

18. Mishra Bhishagratna Brahmashankara, editor, 5th. Chakradatta of Sri

Chakrapanidatta, Vranashothachikitsa: Chapter 44, Verse 40. Varanasi:

Chowkhambha Sanskrit Series Order, 1983; p. 349

19. Mishra Bhishagratna Brahmashankara, editor, 5th. Chakradatta of Sri

Chakrapanidatta, Nadivranachikitsa: Chapter 45, Verse 17, 18. Varanasi:

Chowkhambha Sanskrit Series Order, 1983; p. 361


Vranashothachikitsa: Chapter 47, Verse 40. Varanasi: Chaukhambha

Surbharati Prakashan, 2016; p. 822





Vranashothachikitsa : Chapter 47, Verse 63. Varanasi: Chaukhambha





REFERENCES


Page 174




25. Sastry. J.L.N. Dravyaguna Vijnana, 1st ed. Varanasi: Chaukhambha Orientalia;

2014. p. 152

26. Hegde Prakash, A Harini. A text bok of dravyaguna vijnana, Vol 2. 1st ed.

New Delhi: Chaukhambha publication; 2014. p. 903

27. Chunekar KC. Bhavaprakasha Nighantu of Bhavamishra, 2nd ed. Varanasi:

Chaukambha Krishnadas Academy; 2013. p. 62 - 4

28. Ram Prasad.P. V. V. Madanapala Nighantu. Bombay: Khemaraj

Srikrushnadas Prakashan; 1990. p.16

29. Kamat SD. Studies on Medicinal Plants & Drugs in Dhanvantari Nighantu. 1st

ed. Delhi: Chaukamba Sanskrit Pratishthan. p. 52

30. Pandey G, Dwivedi RR, Baghel MS. Sodala Nighantu of Sodala. 1st ed.

Varanasi: Chaukhambha Krishnadas Academy. p. 39

31. Sharma GP, Sharma PV. Editor. Kaiyadeva Nighantu. Varanasi: Chaukambha

Orientalia. p. 22, 209

32. Tripati ID, Dwivedi V. Editor. Raja Nighantu. 1st ed. Varanasi: Krishnadas

academy. p. 164



REFERENCES


Page 175

34. Pharmacognosy of Ayurvedic drugs, 3rd ed. Trivandrun: Central Research

Institute Tvm; 1998. p. 27- 33

35. The Ayurvedic Pharmacopoeia of India; Ministry of Health and Family

Welfare, Department of Health, Govt of India, Vol1; 1st ed. p. 127- 8

36. R. K. Sharma. Charaka Samhita, 2nd ed. Varanasi: Chowkhambha Sanskrit

Series Office; 2014. p. 14, 18- 38, 55-100

37. Astanga Hrdyam, Sootra sthana, 1st ed. Varanasi: Chowkhambha Sanskrit

Series Office; 2009. p. 339- 70

38. Sushruta Samhita, Sootra sthana, 1st ed. Varanasi: Chowkhambha Sanskrit

Series Office; 2008. p. 41- 2

39. Chunekar KC. Bhavaprakasha Nighantu of Bhavamishra.Varanasi:





Varanasi: Chaukhambha Krishnadas Academy. p. 39




academy. p. 164



REFERENCES


Page 176

45. Kirtikar K R, Basu BD. Indian Medicinal Plants, Vol 1, 2nd ed. Dehradun:

International Book Distributors; 2008. p. 727


Ayurveda Vol3.2nd . New Delhi: Central Counsil for Research in Ayurveda &

Siddha; 2005: p.561- 6

47. Gupta A.K, Tandon Neeraj, Sharma Madhu. Quality standards of Indian

Medicinal Plants. New Delhi: Indian Council of Medical Research ;

2006;Volume 9, p. 179 - 80

48. G. Bapalal. Nighantu Adarsha, 1st ed. Varanasi: Chaukhambha Visvabharati;

1989. Vol 1 p. 435- 40




academy. p. 164

51. Chunekar KC. Bhavaprakasha Nighantu of Bhavamishra. Varanasi:


52. Kamat SD. Dhanvantari Nighantu. 1st ed. Delhi: Chaukamba Sanskrit

Pratishthan; p 52

53. Sastry. J.L.N. Madanapala Nighantu, 2nd ed. Varanasi: Chaukhambha

Orientalia; 2010. p. 59- 62

54. The Ayurvedic Pharmacopoeia of India; Ministry of Health and Family

Welfare, Department of Health, Govt of India, Vol 8; 1st ed. p. 127- 8

REFERENCES


Page 177

55. Agrotechniques of selected medicinal plants. New Delhi: National Medicinal

Plant Board; 2008. p. 175

56. Chaudhuri. A.B. Endangered Medicinal Plants. New Delhi: Daya Publishing

House; 2014: p. 111

57. Srivastava. M , et.al. In vitro conservation of Glycyrrhiza glabra by slow

growth culture. International Journal of Bio- Technology & Research. (IJBTR)

2013; Vol 3(1): p. 49- 58

58. Sukhdev. Prime Ayurvedic Plant Drugs, 2nd ed. New Delhi: Ane Book Pvt.

Ltd.; 2012. p. 365

59. Sheetal Vispute. Glycyrrhiza glabra Linn – Klitaka: A review. International

Journal of Pharma and Bio Sciences 2011; 2(3): 42- 51

60. Toxiocology and Clinical Pharmacology of Herbal Products. New Jersey:

Humana Press; 2000. p. 223 – 30

61. Sastri Haragovinda. Amarakosa of Amarasimha. Varanasi: Chowkhambha

Sanskrit Series Office; 6th ed. 1998: p. 159

62. Jadavaji Trikamji Acharya. Caraka Samhita by Agnivesa. Revised by Charaka

and Dridhabala of Chakrapanidatta. Varanasi: Chaukhambha Sanskrit

Sansthan; 2001

63. Kaviraja Ambikadutta Shastri. Sushruta Samhita edited with Ayurveda Tattva

Sandipika. Varanasi: Chaukhambha Sanskrit Sansthan; 2003

64. Murthy KRS. Astanga hridaya of Vagbhata. 2nd ed. Varanasi: Krishnadas

Academy; 1994.

REFERENCES


Page 178


Chaukambha Krishnadas Academy; 2018. p. 105


Orientalia; p. 198

67. Tripati ID. Dwivedi V. Editor. Raja Nighantu. 1st ed. Varhnadas academy. p.

177 - 178




Pratishthan; p 242


Srikrushnadas Prakashan; 1990. p. 38

71. Sri Shaligram Vaishya. Shaligrama Nighantu Bhushanam. Bombay: Kemraj

Krishnadas Prakashan; 1995. P. 151 2


Varanasi: Chaukhambha Krishnadas Academy; 2009. p. 242





2006;Volume 3, p. 367

REFERENCES


Page 179

75. Jadavaji Trikamji Acharya. Caraka Samhita by Agnivesa. Revised by Charaka

and Dridhabala of Chakrapanidatta. Varanasi: Chaukhambha Sanskrit

Sansthan; 2001

76. Kaviraja Ambikadutta Shastri. Sushruta Samhita edited with Ayurveda Tattva

Sandipika. Varanasi: Chaukhambha Sanskrit Sansthan; 2003

77. Murthy KRS. Astanga hridaya of Vagbhata. 2nd ed. Varanasi: Krishnadas

Academy; 1994.

78. Murthy KRS. Astanga sangraha of Vagbhata. 9th ed. Varanasi: Chaukhamba

Orientalia; 2005.


Orientalia; p. 198

80. Tripati ID. Dwivedi V. Editor. Raja Nighantu. 1st ed. Varanasi: Krishnadas

academy. p. 177- 8




Pratishthan; p 242



84. Sri Shaligrama Vaishya. Shaligrama Nighantu. Bombay: Kemraj Krishnadas

Prakashan; 1995.p. 151- 2

REFERENCES


Page 180


Varanasi: Chaukhambha Krishnadas Academy; 2009. p. 242



87. Kirtikar K R, Basu BD. Indian Medicinal Plants, 2nd ed. Dehradun:

International Book Distributors; 2008. Vol 2. p. 1074- 5

88. Kirtikar K R, Basu BD. Indian Medicinal Plants, 2nd ed. Dehradun:

International Book Distributors; 2008. Vol 2. p. 1074- 5

89. Dey Dipali, Das M N, Sharma A K, O P Gupta editor , Pharmacognosy of

Indigenous Drugs, New Delhi: Central Council for Research In Ayurveda and

Siddha; 2005. Vol 3. p. 1163



Siddha; 2005: p. 206



Siddha; 2005: p. 207- 8



2006;Volume 3, p. 369

REFERENCES


Page 181



Siddha; 2005: p. 209 - 10



2006;Volume 3, p. 372

95. SharmaP.C, Yelne M.B, DennisT.J. Database on Medicinal Plants Used in


Siddha; 2005: p. 209



2006;Volume 3, p. 372

97. Sushruta. Chi 1/6. In: Y T Acharya (eds.) Sushruta Samhita. Varanasi:

Chaukhambha Publications; 2006. p. 396

98. Sushruta. Su 21/40. In: Y T Acharya (eds.) Sushruta Samhita. Varanasi:








REFERENCES


Page 182



103. Sushruta. Su 23/18, Chi 1/7. In: Y T Acharya (eds.) Sushruta Samhita.

Varanasi: Chaukhambha Publications; 2006. p. 396

104. Vagbhata. Ut 25/11. In: Paradakara HS (eds.) Ashtanga Hridaya. 3rd ed

Varanasi: Chaukhambha Orientalia; 2011. p. 865

105. Agnivesha. Chi 25/26. In: Y T Acharya (eds.) Charaka Samhita. Varanasi:

Chaukhambha Publication; 2006. p. 593

106. Madhavakara. Vol2. In: Brahmanand Tripathi (eds.) Madhavanidana. 2nd

ed. Varanasi: Chaukhambha Surbharati Prakashan; 1998. p. 275





109. Agnivesha. Chi 25. In: Y T Acharya (eds.) Charaka Samhita. Varanasi:


110. Vagbhata. Ut 29/4. In: Gupta Kaviraj Atrideva (eds.) Ashtanga Sangraha.

3rd ed. Varanasi: Chaukhambha sanskrit Series office; 1993.





REFERENCES


Page 183

113. Vagbhata. Ut 25/2-4. In: Paradakara HS (eds.) Ashtanga Hridaya. 3rd ed.


114. Madhavakara. Vol 2. In: Brahmanand Tripathi (eds.) Madhavanidana. 2nd

ed. Varanasi: Choukamba Surbharati Prakashan; 1998. p. 275

115. Sharangadhara. Pu kh 7/71-74. In: Dr. Shailaja Sreevasthava (eds.)

Sharangadhara Samhita, Jeevan Prada Vyakhya. 2nd ed. Varanasi:

Chaukhambha Samskruta Samsthana. p. 265















REFERENCES


Page 184







126. Sushruta. Su 23/3-5. In: Y T Acharya (eds.) Sushruta Samhita. Varanasi:




128. Sushruta. Su 23/8. In: Y T Acharya (eds.)Sushruta Samhita. Varanasi:










133. Sushruta. Chi1/13. In: Y T Acharya (eds.) Sushruta Samhita. Varanasi:


REFERENCES


Page 185



135. Vriddha Jeevaka. Dvivrana Chikitasa adhyaya. In: Nepalarajagruna Pandit

Hemaraj Shrama (eds.) Kashyapa Samhita. 7th ed. Varanasi: Chaukhambha

Samskruta Samsthana; 2000

136. Vagbhata. Ut 25/2-4. In: Paradakara HS (eds.) Ashtanga Hridaya. 3rd ed.




138. Hamilton Bailey & McNeill Love. In: Russell R.C.G, Williams Norman S

& Bulstrode Christopher J.K (eds.) Short Practice of Surgery. 24th ed. London:

Hodder Arnold Publication;2004

139. Somen Das. Ulcer, sinus and fistula. A Concise Textbook of Surgery. 6th

ed. Calcutta: S Das Publications;2004. p. 156

140. Somen Das. Ulcer, sinus and fistula. A Concise Textbook of Surgery. 6th

ed. Calcutta: S Das Publications;2004. p. 158

141. Hamilton Bailey & McNeill Love. In: Russell R.C.G, Williams Norman S

& Bulstrode Christopher J.K (eds.) Short Practice of Surgery. 24th ed. London:

Hodder Arnold Publication;2004

142. https: // life.umd.edu/ classroom/ bsci

143. Chakraborty P. A Textbook of Microbiology. Kolkata: New Central Book

Agency (P) LTD; 2009: p. 238- 46

REFERENCES


Page 186


Agency (P) LTD; 2009: p. 316- 22


Agency (P) LTD; 2009: p. 343- 46


Agency (P) LTD; 2009: p. 348 - 49

147. Gerard. J. tortora, Berdell. R. Funke; Microbiology an introduction, 12ed.

Pearson publication; 2016. p. 160- 1

148. M. Joanne, M. Hinda, J. Christopher willey; Prescott’s microbiology, 9th

ed. Mcgraw hill publications; 2014. p. 154


ed. Mcgraw hill publications; 2014. p. 158

150. B.S. Nagoba, Asha pichare; Medical microbiology, 2nd ed. Elsevier

Publications, 2014. p. 46- 7


Publications, 2014. p. 115


ed. Mcgraw hill publications; 2014. p. 160- 2

153. Ananthanarayan. R. Jayaram Panicker C K J. Medical mycology. In: (eds.)

Textbook of Microbiology. 7th ed. Madras: Orient Longman; 2005. p. 19


Publications, 2014. p.16

REFERENCES


Page 187

155. Gerard. J. tortora, Berdell. R. Funke; Microbiology an introduction, 12ed.

Pearson publication; 2016. p. 187

156. Trease, Evans; Pharmacognosy, 15th ed . W B Saunders Publication. 2005.

p. 98

157. Saxena Sanjai. Glycyrrhiza glabra: Medicine over the millennium. Natural

Product Radiance. (NPR) 2005; vol 4(5) : p. 358 – 67

158. BadkhaneYogesh,Yadav A.S, Bajaj. A, Sharma. AjitK,Raghuwanshi D.K.

Glycyrrhizaglabra L. A miracle medicinal herb . Indo American Journal of

Pharmaceutical Research. 2014; 4(12): p. 5808- 16

159. Sharma P.V Dravyaguna Vijnana Vol 2. Varanasi: Chaukhambha Bharati

academy; 2015: p. 253

160. Sharma P.V Dravyaguna Vijnana Vol 2. Varanasi: Chaukhambha Bharati

academy; 2015: p. 472

161. Dhyani SC. Rasa Panchaka. Varanasi: Chowkhambha Krishnadas

Academy; 2016

162. Hisanori Akiyama,et.al. Antibacterial action of several tannins against

Staphylococcus aureus, Journal of Antimicrobial Chematherapy, 2001

163. Augustin Scalbert. Antimicrobial properties of tannins, Phytochemistry,

1991

164. Nouioua wafa, et.al. The antioxidant and antimicrobial activities of

flavonoids and tannins extracted from Phlomis bovei De Noe, European

Journal of Experimental Biology, 2016

REFERENCES


Page 188

165. Michal Arabski, et.al. Effect of Saponins against clinical E.coli strains and

Eukaryotic cell line, Journal of Biomedicine and Biotechnology, 2012

166. Joshita Sabbineni. Phenol- An efffective antibacterial agent, Journal of

Medical & Organic Chemistry, 2016

167. WD Armstrong. Antibacterial effects of Quinones, SAGE journals, 1943

168. Paul M Borick. Antimicrobial activity of some higher Amine Salts of

Carboxylic acids, Applied Microbiology, 1961

169. Escoula L. Effeect of carbohydrates on the antbacterial activity of patulin

on Steptococcus bovis, Pubmed , 1982

170. Sugars in human mother's milk are non toxic antibacterial agents,

Vanderbilt institute, 2017

171. Killeen, G., Madigan, C., Connolly, C., Walsh, G., Clark, C., Hynes, M.,

Timmins, B., James, P., Headon, D., Power, R., 1998. Antimicrobial saponins

of Yucca schidigera and the implications of their in vitro properties for their in

vivo impact. J. Agric. Food Chem., 46(8):3178- 86. [doi:10.1021/jf970928j]

172. Mshvildadze, V., Favel, A., Delmas, F., Elias, R., Faure, R., Decanosidze,

Q., Kemertelidze, E., Balansard, G., 2000. Antifungal and antiprotozoal

activities of saponins from Hedera colchica. Pharmazie, 55(4):325- 6.

173. Cowan M., 1999. Plant products as antimicrobial agents. Clinical

Microbiology Review, 12(4):564- 82.

174. Tsuchiya, H., Sato, M., Miyazaki, T., Fujiwara, S., Tanigaki, S., Ohyama,

M., Tanaka, T., Iinuma, M., 1996. Comparative study on the antibacterial

REFERENCES


Page 189

activity of phytochemical flavanones against methicillin-resistant

Staphylococcus aureus. J. Ethnopharmacol., 50(1):27-34.

175. Omulokoli, E., Khan, B., Chhabra, S., 1997. Antiplasmodial activity of

four Kenyan medicinal plants. J. Ethnopharmacol., 56(2):133- 7.

[doi:10.1016/S0378-8741(97)01521-3]

176. Phillipson, J., O′Neill, M., 1987. New leads to the treatment of protozoal

infections based on natural product molecules. Acta Pharm. Nord., 1:131- 44.

177. Scalbert Augustin. Antimicrobial properties of Tannins. Phytochemistry,

Vol. 30, No. 12. pp. 3875- 83, 1991

178. .R .Balabhaskar, K.Vijayalakshmi. High-Performance Thin-Layer

Chromatography Fingerprint Profile Of Bauhinia Tomentosa Linn.Leaves.

Asian Journal of Pharmaceuticle and Clinical Research, Vol 11, Issue 2, 2018

179. Debalke D, Birhan Mastewal, et. al. Assessments of antibacterial effects of

aqueous- ethanolic extracts of Sida rhombifolia’s aerial part. Hindawi

Scientifific World Journal Volume 2018, Article ID 8429809, 8 pages

https://doi.org/10.1155/2018/8429809

180. B.S. Nagoba, Asha pichare; Medical microbiology & Parasitology, 3rd ed.

Elsevier Publications. p.103 - 9

https://doi.org/10.1155/2018/8429809

DEPT OF DRAVYA GUNASRI DHARMASTHALAMANJUNATHESHWARA COLLEGE OF AYURVEDA &

HOSPITAL, HASSAN

ANTIBACTERIAL ACTIVITY OF DHATAKI (WOODFORDIA FRUTICOSA (L.) KURZ) ASPRATINIDHI DRAVYA FOR YASHTIMADHU (GLYCYRRHIZA GLABRA LINN) WITH

REFERENCE TO VRANAHARA KARMA

PG SCHOLAR: Dr. IBAMEAIMON POHTAM GUIDE: Dr. ANURADHA KNCO-GUIDE: Mrs. SHASHIREKHA KS

SCREENING FORM /CASE PROFORMA

(Enter a � in the appropriate box)

1. OPD No. :2. Screening Subject Sl. No. :3. Name of the Subject :4. Gender : Male ( ), Female ( )5. Age :6. Address :7. Telephone No. :

8. Diagnosis and Inclusion Criteria

Yes No1. Complaints of wound of long duration2. Complaints of wound with discharge3. Complaints of Burning sensation4. Complaints of Swelling5. Complaints of Itching6. Willing and able to participate and ready to sign consent form

9. Criteria for ExclusionYes No

1. Patient with H/O or presenting with Tuberculosis2. Patient with H/O or presenting with Malignancy3. HIV, HBsAg positive patient4. Patients with any complications which may interfere with the

course of study

Remark : Whether patient/ participant is suitable for enrolment into study ? Yes / NoIf Enrolled : - Subject Enrollment No. :

Sign of Scholar Sign of Guide Sign of Co-Guide

DEPT OF DRAVYA GUNASRI DHARMASTHALAMANJUNATHESHWARA COLLEGE OF AYURVEDA &

HOSPITAL, HASSAN

ANTIBACTERIAL ACTIVITY OF DHATAKI (WOODFORDIA FRUTICOSA (L.) KURZ) ASPRATINIDHI DRAVYA FOR YASHTIMADHU (GLYCYRRHIZA GLABRA LINN) WITH

REFERENCE TO VRANAHARA KARMA

PG SCHOLAR: Dr. IBAMEAIMON POHTAM GUIDE: Dr. ANURADHA KNCO-GUIDE: Mrs. SHASHIREKHA KS

OPD No. / IPD No. :Subject Sl. No. :Name of the Subject :Gender : Age:Address :

CULTURE AND SENSITIVITY TESTTest Sample: PUS

Date of Sample collection :

Body area from where sample is collected :

Date and Time of commencement culture and sensitivity :

Culture Observation

Sample Incubation

period

Growth on

MacConkey

Colony

Morphology

Microscopy

by Gram

Staining

Organism

Identified

PUS 37℃ for 24

hours

Growth Seen

SENSITIVITY TEST OBSERVATION:

ORGANISM :

AQUEOUSEXTRACT

CONCENTRATION

DHATAKI(Woodfordia fruticosa)

YASHTIMADHU(Glycyrrhiza glabra)

SENSITIVITY RESISTANCE SENSITIVITY RESISTANCE600 µl

500 µl

400 µl

300 µl

200 µl

100 µl

Sign of Scholar Sign of Guide Sign of Co-Guide

Master chart

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz) as pratinidhi dravya for Yashtimadhu(Glycyrrhiza glabra Linn) with reference to vranahara karma”

Page 1

SubjectNo.

Age Gender Deerghakaleena

Puyasrava Daha Kandu Shotha Organisms

146 - 60 Female Present Present Absent Absent Present


261 - 75 Female Present Present Absent Absent Absent






561 - 75 Male Present Present Absent Absent Present


61 - 15 Male Present Present Absent Present Present


7 76 - 85 Female Present Present Present Present Absent Escherichia coli

8 61 - 75 Male Present Present Present Absent Absent Escherichia coli


1046 - 60 Female Present Present Present Absent Absent


11 31 - 45 Male Present Present Present Present Absent Escherichia coli

12 46 - 60 Female Present Present Present Absent Absent Escherichia coli

Master chart


Page 2

1316 - 30 Male Present Present Present Present Absent


1446 - 60 Male Present Present Absent Absent Absent


15 61 - 75 Male Present Present Absent Absent Absent Escherichia coli

1631 - 45 Male Present Present Present Present Present







2061 - 75 Female Present Present Present Absent Present


2146 - 60 Male Present Present Present Absent Present




23 61 - 75 Female Present Present Present Absent Present Escherichia coli



Master chart


Page 3



26 16 - 30 Male Present Present Present Absent Present Escherichia coli

27 61 - 75 Male Present Present Present Absent Present Acinetobacter

28 31 - 45 Female Present Present Absent Absent Absent Acinetobacter



31 46 - 60 Male Present Present Present Present Absent Acinetobacter

3246 - 60 Female Present Present Present Absent Present


33 46 - 60 Male Present Present Absent Absent Present Escherichia coli


3531 - 45 Male Present Present Present Absent Absent


36 46 - 60 Female Present Present Present Present Present Acinetobacter



3846 - 60 Female Present Present Present Present Absent


Master chart


Page 4



4046 - 60 Male Present Present Present Present Absent


4116 - 30 Female Present Present Present Present Present







45 46 - 60 Male Present Present Absent Present Absent Escherichia coli





49 46 - 60 Male Present Present Present Present Absent Acinetobacter



51 46 - 60 Male Present Present Present Present Present Acinetobacter

Master chart


Page 5



5346 - 60 Female Present Present Absent Present Present


5446 - 60 Male Present Present Present Present Present


55 16 - 30 Female Present Present Present Present Present Acinetobacter

5616 - 30 Female Present Present Present Absent Absent


5776 - 85 Male Present Present Present Absent Absent




60 31 - 45 Male Present Present Present Absent Present Escherichia coli

Master chart


Page 6

Sensitivity and Zone of inhibition (ZOI) – Yashtimadhu - Staphylococcus aureus(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)

Con 600µg/µl Con 500 µg/µl Con 400 µg/µl Con 300 µg/µl Con 200 µg/µl Con 100 µg/µlM in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI

16 S 17 S 15 S 17 S 12 MS 15 S25 S 15 S 16 S 15 S 15 S 12 MS17 S 17 S 19 S 16 S 15 S 14 MS18 S 15 S 13 MS 14 MS 12 MS 0 R18 S 16 S 16 S 18 S 18 S 12 MS15 S 15 S 15 S 14 MS 14 MS 11 MS21 S 18 S 19 S 17 S 16 S 13 MS17 S 20 S 17 S 17 S 15 S 13 MS28 S 25 S 20 S 20 S 20 S 0 R18 S 18 S 18 S 15 S 16 S 13 MS15 S 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R10 MS 8 R 10 MS 10 MS 10 MS 7 R12 MS 10 MS 0 R 0 R 0 R 0 R16 S 10 MS 10 MS 13 MS 15 S 10 MS0 R 0 R 0 R 0 R 10 MS 10 MS0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R18 S 15 S 13 MS 12 MS 11 MS 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R10 MS 10 MS 10 MS 10 MS 10 MS 0 R15 S 15 S 13 MS 10 MS 0 R 0 R

Master chart


Page 7

Sensitivity and Zone of inhibition (ZOI) – Dhataki - Staphylococcus aureus(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)

Con 600µg/µl Con 500 µg/µl Con 400 µg/µl Con 300 µg/µl Con200 µg/µl Con 100 µg/µl

M in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI M in mm ZOI

20 S 17 MS 15 MS 14 R 22 S 13 R14 R 21 S 20 MS 20 MS 22 S 20 MS26 S 25 S 24 S 28 S 30 S 20 MS25 S 23 S 26 S 22 S 20 MS 18 MS25 S 24 S 30 S 24 S 24 S 28 S22 S 20 MS 21 S 22 S 20 MS 24 S27 S 26 S 24 S 26 S 26 S 22 S25 S 25 S 27 S 23 S 28 S 22 S20 MS 25 S 15 MS 15 MS 13 R 10 R18 MS 17 MS 20 MS 20 MS 17 MS 22 S17 MS 20 MS 18 MS 15 MS 15 MS 13 R15 MS 13 R 15 MS 16 MS 16 MS 13 R17 MS 17 MS 15 MS 13 R 12 R 12 R18 MS 15 MS 15 MS 12 R 10 R 10 R22 S 20 MS 19 MS 24 S 16 MS 15 MS20 MS 20 MS 20 MS 18 MS 13 R 10 R15 MS 15 MS 14 R 13 R 11 R 10 R20 MS 18 MS 15 MS 14 R 15 MS 13 R22 S 20 MS 15 MS 13 R 12 R 12 R20 MS 20 MS 15 MS 13 R 12 R 12 R23 S 20 MS 15 MS 13 R 13 R 13 R24 S 23 S 20 MS 20 MS 15 MS 15 MS23 S 22 S 18 MS 20 MS 16 MS 15 MS23 S 22 S 22 S 20 MS 15 MS 12 R25 S 22 S 22 S 20 MS 17 MS 15 MS

Master chart


Page 8

Sensitivity and Zone of inhibition (ZOI) – Yashtimadhu - Escherichia coli(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)


0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 10 MS 11 S 7 MS 15 S0 R 0 R 35 S 0 R 0 R 0 R0 R 0 R 0 R 0 R 12 S 10 MS0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R12 S 10 MS 13 S 0 R 10 MS 10 MS0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 15 S 25 S0 R 0 R 0 R 0 R 0 R 0 R

Master chart


Page 9

Sensitivity and Zone of inhibition (ZOI) – Dhataki - Escherichia coli(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)


20 S 23 S 25 S 29 S 25 S 24 S22 S 25 S 26 S 21 S 24 S 19 S20 S 20 S 19 S 17 MS 20 S 19 S15 MS 15 MS 16 MS 15 MS 13 MS 11 R30 S 29 S 28 S 25 S 26 S 22 S22 S 22 S 18 S 18 S 16 MS 16 MS21 S 20 S 22 S 20 S 21 S 15 MS20 S 20 S 20 S 17 MS 18 MS 12 R15 MS 20 S 20 S 15 MS 10 R 12 R18 MS 17 MS 15 MS 13 MS 14 MS 12 R18 MS 13 MS 15 MS 16 MS 10 R 10 R18 MS 17 MS 17 MS 14 MS 12 R 12 R12 R 12 R 15 MS 15 MS 13 MS 12 R20 S 15 MS 16 MS 16 MS 13 MS 14 MS25 S 22 S 21 S 20 S 15 MS 15 MS25 S 20 S 18 MS 15 MS 15 MS 15 MS25 S 20 S 20 S 20 S 20 S 12 R20 S 20 S 18 MS 18 MS 15 MS 10 R

Master chart


Page 10

Sensitivity and Zone of inhibition (ZOI) – Yashtimadhu - Acinetobacter(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)


0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R20 S 15 S 10 MS 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R0 R 0 R 0 R 0 R 0 R 0 R13 S 13 S 15 S 0 R 0 R 0 R18 S 15 S 0 R 0 R 0 R 0 R10 MS 10 MS 10 MS 10 MS 10 MS 0 R

Sensitivity and Zone of inhibition (ZOI) – Dhataki - Acinetobacter(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)


18 MS 17 MS 15 MS 15 MS 14 MS 11 R18 MS 18 MS 15 MS 15 MS 13 MS 12 R13 MS 17 MS 16 MS 17 MS 13 MS 10 R18 MS 20 S 17 MS 15 MS 15 MS 12 R22 S 18 MS 15 MS 14 MS 12 R 10 R25 S 25 S 23 S 23 S 17 MS 12 R23 S 23 S 22 S 20 S 20 S 11 R25 S 25 S 21 S 18 MS 16 MS 14 MS20 S 20 S 20 S 18 MS 15 MS 15 MS

Master chart


Page 11

Sensitivity and Zone of inhibition (ZOI) – Yashtimadhu - Pseudomonas aeruginosa(Con – Concentration, M- measurement, R- Resistant, MS- Moderately sensitive, S- Sensitive)

Con 600µg/µl Con 500 µg/µl Con 400 µg/µl Con 300 µg/µl Con 200 µg/µl Con 100 µg/µl

M in mm ZOI M inmm

ZOI M inmm

ZOI M in mm ZOI M in mm ZOI M in mm ZOI

0 R 0 R 0 R 0 R 0 R 0 R

0 R 0 R 0 R 0 R 0 R 0 R

10 MS 10 MS 10 MS 12 S 7 MS 7 MS

15 S 18 S 0 R 0 R 0 R 0 R

0 R 0 R 0 R 0 R 0 R 0 R

0 R 0 R 0 R 0 R 0 R 0 R

0 R 0 R 0 R 0 R 0 R 0 R

16 S 13 S 12 S 10 MS 10 MS 10 MS

Master chart


Page 12

Annexures

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz.) as pratinidhi dravyafor Yashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranahara karma”

Page 190

Figure 7.Macroscopy of crude drug Yashtimadhu (Glycyrrhiza glabra Linn.)

Annexures


Page 191

Figure 8: Microscopy of Yastimadhu root

Fig 8a. T.S of root

Fig 8b. A portion enlargedCa – cambium; Ck – cork; Ct – cortex; Pa – parenchyma; Perd – periderm PF – phloemfibres; Ph – phloem; Ve – vessel; XF – xylem fibres; XR – xylem ray.

Ck↓

Perd→

Ph→

Ve→XF

Perf→

PF→

PF→

Ct→

Pa→

Ck↓

Ca→

Ph→

Annexures


Page 192

Fig 8c. Cork, periderm,cortex

Fig 8d.PhloemCk – cork; Ct – cortex; CF–crystal fibres; ID–idioblasts; MR–medullary rays; Pa –parenchyma; Perd – periderm; Perf – pericyclic fibres; Ph – phloem; SG – starch grains.

Perd→

Ph→

SG→

Perf→

Ct→

ID→

CF→

MR→

Ck→

Pa→

Annexures


Page 193

Fig 8e.Phloem,cambium,xylem

Fig 8f.Cambium and xylem

Fig 8g.Xylem and pithCa–cambium; Ph–phloem; Phf – phloem fibres; Ve – vessel; Xy–xylem; XF – xylemfibres; XR – xylem ray.

Ca→

Xy→

Pi→

XR→

Xy→

Ve→

XF→

Ph→

Phf→

Ca→

XR→

Annexures

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz.) as pratinidhi dravya forYashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranahara karma”

Page 194

Fig 8h.Cork , periderm and cortex withstarch grains

Fig 8i. Stratified cork and cortex

Fig 8j.Xylem Fig 8k.Phloem fibresCk-cork; Ct–cortex; Perd–peroderm; Ph–phloem; Phf – phloem fibres; SG–starch grains; Ve –vessel; Xy–xylem; XF – xylem fibres; XR – xylem ray.

XF→

Ve→

XR→PF→

SG→

Ck↓

Perd→

Ph→

Ve→

Ct→

Ck↓

Ct→

Annexures

“Antibacterial activity of Dhataki (Woodfordia fruticosa (L.) Kurz. As pratinidhidravya for Yashtimadhu (Glycyrrhiza glabra Linn.) with reference to vranaharakarma ”

Page 195

Figure 9. Macroscopy of flower of Dhataki (Woodfordia fruticosa (L.) Kurz.)

Annexures


Page 196

. Figure 10: Microscopy of corolla of Dhataki

Fig 10a. TS of corolla

Fig 10b. A portion enlargedCu – cuticle; LE – lower epidermis; Me – mesophyll; Pal – palisade cells; SP –spongy parenchyma; T –trichome; UE – upper epidermis; VB – vascular bundle.

Pal→

UE→

Me→

SP→

UE→

VB→

Cu→

VB→

LE→

Pal→

Cu→

T→

LE→

Annexures


Page 197

Figure 11: Microscopy of Anther

Fig 11a. TS of anther

Fig 11b. Pollen grainsEp – epidermis; Po – pollen sac with pollen grains; SC – sclerified layer.

Po→

Po→

Ep→

SC→

Annexures


Page 198

Figure 12: Microscopy of Ovary

TS of flower through ovaryE–epidermis; FW–fruit wall; T–trichomes; VB–vascular bundle

Figure 13. TS of stalkCol – collenchymas; Ct – cortex; Cu – cuticle; Ep – epidermis; Pa – parenchyma; Ph –phloem; Pi – pith; RP – reticulated parenchyma; T – trichome; Ve – vessels; Xy – xylem.

VB→

FW→

E→

T→

T→

Ve→

Col→

E→

Cu→

RP→

Pi→Xy→Pa→

Ph→

Annexures


Page 199

Figure 14. Collection of Dhataki

Fig 15. Dhataki - Coarse powder Fig 16. Yashtimadhu - Coarse powder

Annexures


Page 200

Fig 17.Soxhlet apparatus- Yashtimadhu Fig 18. Soxhlet apparatus- Dhataki

Fig 19. Water bath Fig. 20 Extracts of Dhataki&Yashtimadhu

Annexures


Page 201

Fig 21. Phytochemical analysis

Fig 22. Loss on drying- Dhataki Fig 23. Loss on drying -

Yashtimadhu

Annexures


Page 202

Fig 24. Water & alcohol extractive value

Fig 25. Ash value

Annexures


Page 203

Fig 26. Preparation of Media

Fig 27. Serial Dilution - Concentration

Annexures


Page 204

Fig 28. Collection of pus

sample

Fig 29. Culturing of Bacteria

Fig 30. Sensitivity Test

Annexures


Page 205

Fig 31. Sensitivity Test

Annexures


Page 206

Fig 32. Zone of inhibition of Dhataki

Fig 33. Zone of inhibition of Yashtimadhu

Annexures


Page 207

Fig 34. Measurement of Zone of inhibition

Annexures


Page 208

Annexures


Page 209

Annexures


Page 210

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