GJRMI - Volume 1, Issue 3 - March 2012

www.gjrmi.com GJRMI, Volume 1, Issue 3, March 2012, 62 - 68

Global Journal of Research on Medicinal Plants & Indigenous Medicine



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Dr Hari Venkatesh K Rajaraman M.D.(Ay) P.G.D.H.M

Advisory Board

Prof. Rabinarayan Acharya M.D., Ph.D (Ay)

Dr. Mathew Dan M.Sc., Ph.D (Botany)

Dr. Tanay Bose M.Sc., (Ph.D) (Botany)

Dr. Nagaraja T.M M.Pharm.,Ph.D (Pharm)

Dr. Narappa Reddy M.D. (Ay)

Editorial board

Dr. Kumaraswamy M.Tech. Ph.D (Bio-Tech)

Dr. Madhu .K.P M.D. (Ay)

Dr. Sushrutha . C.K M.D.(Ay)

Dr. Ashok B.K

Dr. Janardhana.V.Hebbar M.D. (Ay)

Dr. Shwetha Hari Venkatesh B.A.M.S.

Dr. Vidhya Priya Dharshini. K.R. B.N.Y.S.

Mr. R. Giridharan M.Tech (Bio-tech)

Miss. Shyamala Rupavahini M.Sc., M.Phil (Bio-Chem)

Honarary Members - Editorial Board

Dr. Shubha Ganguly M.V.Sc.,Ph.D

Dr Farhad Mirzaei, M.Sc., Ph.D.,



Original Research Article

MALARIA: NOVEL PLANT REMEDIES SHOW GREAT PROMISE IN

TREATING THE DEADLY DISEASE

Taba K.M.*†, Paulus J.

**, Kayembe J.S.

*

* University of Kinshasa, Faculty of Sciences, Dept of Chemistry P.O. Box 190 Kinshasa XI, Democratic Republic of

Congo

** University of Kinshasa, Faculty of Sciences, Dept of Biology P.O. Box 190 Kinshasa XI, Democratic Republic of

Congo

† Corresponding Author - Tel. +243 81 333 02 42 [email protected]

Received : 13/01/2012; Revised : 17/02/2012; Accepted : 27/02/2012;

ABSTRACT

Clinical investigation of eight plant remedies used as traditional medicines in Kinshasa, D.R. Congo,

to treat Malaria patients showed significant removal of parasites in the blood, as well as elimination

of clinical detection of disease. The percentage recovery from Malaria depends on the type of

remedy chosen: Cassia occidentalis Linn. (97%), Carica Papaya Linn. (94%), Cymbopogon

citratus (DC.) Staff, 1906 (93%), Garcinia kola Heckel. (94%), Lantana camara L. (90%), Ocimum

gratissimum L. (86%), Phyllanthus niruri L. (93%) and Vernonia amygdalina Delile. (67%). No

identifiable side effects were noticed during and after treatment. In vitro study of alcohol extracts of

these remedies showed an inhibition concentration of Plasmodium growth of 83% (12.5 µg/ml) for

C. occidentalis Linn., 91% (25 µg/ml) for C. Papaya Linn., 93% (25 µg/ml) for C. citratus (DC.)

Staff, 1906, 93% (25 µg/ml) for L. camara L., 100% (12.5 µg/ml) for O. gratissimum L., 100%

(12.5µg/ml) for P. niruri L. and 0% (25 µg/ml) for V. amygdalina Delile. Secondary metabolites

were isolated from some plant remedies and their IC50 determined in vitro on P. falciparum. The

highest IC50 was observed in alkaloid extract of P. niruri L. (8 µg/ml); terpenes extract of O.

gratissimum L. (0.27µg/ml); quinone extract of G. kola Heckel. (1.02 µg/ml) and flavonoid extract

of G. kola Heckel. (1.5µg/ml).

Keywords: Malaria, Plant Remedies, Cassia occidentalis Linn.,Carica Papaya Linn.,

Cymbopogon citratus (DC.) Staff, 1906, Garcinia kola Heckel., Lantana camara L., Ocimum

gratissimum L., Phyllanthus niruri L., Vernonia amygdalina Delile.



INTRODUCTION

Malaria constitutes a major health problem to

wide populations across the tropical and

subtropical areas of the world. Due to

increasing levels of drug resistance and

unaffordable prices, many poor areas in Africa

cannot access the newer chemotherapeutic

pharmaceuticals (Jurg et al., 1991). For

thousands of years, traditional medicines have

been used to treat Malaria. Some herbal

remedies have been the sources of two main

groups of modern Anti-malarial drugs. Quinine

from Cinchona species (Andrade-Neto et al.,

2003) and Artemisinin from Artemisia species

(Anonym, 1982; Li and Rieckmann, 1992;

Wang and Xu, 1985). Quinine and Artemisinin

have also been used as template molecules in

the synthesis of other Anti-malarial drugs such

as Amodiaquine from Quinine (Chen et al.,

1987; O’Neil et al., 2003) as well as

Artemether and Artesunate from Artemisinin

(Pe et al., 1989; Yang, Ski, Li, 1982).

Collaborations with traditional healers are

needed so that we may understand the use of

ancestral medicines in modern practice.

Knowledge about efficacy and safety of herbal

treatments provides the possibility of

combating these diseases at primary healthcare

level, and will also provide new leads for

research on new Malaria therapies. Based on

this, it was of high importance to access the

efficacy, both clinically and in-vitro, of several

antimalarial remedies used by traditional

healers (Jurg et al., 1991).

An ethno-botanic survey conducted in gardens

in the suburb of Kinshasa, the capital of D.R.

Congo, led to identification of 58 species of

plants used for treatment of fever (Malaria)

(Ngalamulume et al., 1982; Kasuku et al.,

1999). The area surveyed is the poorest urban

environment of the city where most people are

inclined to use traditional remedies to treat

diseases. The eight most used remedies are:

Cassia occidentalis Linn., Carica papaya

Linn., Cymbopogon citratus (DC.) Staff, 1906,

Garcinia kola Heckel., Lantana camara L.,

Ocimum gratissimum L., Phyllanthus niruri

and Vernonia amygdalina Delile.

The current work focuses on the Anti-malarial

activity of these remedies clinically by

replicating the traditional healer’s therapeutic

procedure in clinical trials. Furthermore, the in

-vitro inhibition of Plasmodium falciparum

growth with crude extracts is also being

evaluated and documented.

Experimental

Preparation of materials

Eight plant materials were collected by

traditional healers in the suburb of Kinshasa.

These were authenticated at Herbarium of the

University of Kinshasa, where specimens are

deposited. The decoction of each plant (except

G. kola Heckel. of which seed is chewed) was

prepared according to the traditional healer

protocol. The dose and the mode of

administration proposed by traditional healer

are reported in Table 3.

200g of dried plant material was taken up with

1 liter of ethanol and refluxed for 6 hours.

Upon cooling, ethanol was removed under

reduced pressure. The secondary metabolites,

Alkaloids, Terpenes, Quinones and Flavonoids

were obtained according to the classical

procedure described by Bruneton (Bruneton,

1993). Thin layer chromatography analysis /

preparation of extracts and fractions were

performed on pre-coated plates (Silica gel 60

F254, MERCK). The spots were observed under

UV (254 and 366 nm).

Clinical investigation

Patients were treated with each traditional

remedy according to the protocol of traditional

healers at modern health centre. Patients’

selection was based on: (1) indications of

malaria and of parasites P. falciparum

trophozoites in Giemsa stained film prepared

from capillary blood; (2) the absence of Anti-

malarial drug use; (3) age between 1 and 60

years (Pregnant women and severe sick patients

were excluded) (Jurg et al., 1991).

All external signs of malaria including fever,

Sweating, Vomiting, Nausea, Headache,

Muscle and Joint pains, were recorded before



and during treatment. Blood samples were also

taken at the starting and at days 5, 7, 9 and 11

of treatment. Asexual parasitaemia was

quantified by counting against white blood

cells (number of asexual parasites/500

leucocytes × 8000 leucocytes/µl), (Jurg et al.,

1991). The results of clinical investigation are

reported in Table 1.

Table 1: Results of clinical investigation and in-vitro inhibition of Plasmodium’s growth by total

alcohol extracts

Plants Clinical investigation Alcohol extracts

No. of

Patients

% of

recovery

Conc (µg/ml) % of inhibition

C. citratus (DC.) Staff, 1906

C. occidentalis Linn.

C. papaya Linn.

G. kola Heckel.

L. camara L.

O. gratissimum L.

P. niruri L.

V. amygdalina Delile.

Chloroquine

56

62

46

39

50

69

83

40

93

97

94

94

90

86

93

67

25

12.5

25

12.5

25

12.5

ND*

25

12.5

12.5

12.5

25

0.1

93

53

83

24

91

20

ND*

93

0

100

100

0

100

ND*: Not determined

Test for in-vitro Anti-malarial activity

The in-vitro assays were conducted by using

the micro dilution technique of Desjardin

(Desjardin et al., 1979). The P. falciparum

parasites were derived by direct visualization

and micro manipulation from fresh patient

isolates. The test compounds were initially

dissolved in ethanol: water mixture (1:3) or in

Dimethylsulfoxide and diluted 100- fold in

Roswell Park Memorial Institute 1640 (RPMI,

Sigma Aldrich) culture medium, supplemented

with 25mM Hepes and 32mM NaHCO3. These

solutions were diluted in 10 different

concentrations. The parasites were exposed to

different dilutions of each compound for 48h

and incubated at 37°C. Direct estimation of

parasite growth inhibition was used and it was

based on direct reading of smears made in 24-

well, flat-bottomed plates to estimate growth

and evolution stages of the parasites (Benoit et

al., 1996). Parasitaemia and parasite stage were

determined after 48h of contact between

extracts and parasites. Concentration-response

data was analyzed by nonlinear regression

logistic dose response model and IC50 values

for each compound were calculated.

RESULTS AND DISCUSSION

Patients diagnosed with Malaria were treated at

modern health centre via the technique of

traditional healer for a period of 7 days. By the

fifth day, all symptoms of Malaria disappeared

in all cases. The level of Malaria parasite in the

blood however did not completely disappear.

The percentage recovery was calculated based

on both the amount of residual parasite in the

blood, and the elimination of symptoms. The

percentage of recovery ranged from 97% for C.

occidentalis Linn. to 67% for V. amygdalina

Delile. The observed high level of recovery

showed that plant remedies are potential



candidates in the search of new molecules with

potent biological activities (Jurg et al., 1991).

Further, it is necessary to look into classes of

compounds responsible for Anti-plasmodial

activities observed during clinical investigation

and in-vitro studies of these plant remedies.

The most likely classes are Alkaloids,

Terpenes, Quinones and Flavonoids.

Table 2: IC50 values of isolated compounds

Plant species Isolated

compounds

Eluents RF IC50 value

µg/ml

C. papaya Linn.

C. papaya Linn.

Total alkaloids

Alkaloid

-

EtOAc / n -hexane 1:1

-

0.5

185

50

G. kola Heckel.

G. kola Heckel.

G. kola Heckel.

G. kola Heckel.

G. kola Heckel.

G. kola Heckel.

G. kola Heckel.

Quinone

Quinone

Quinone

Quinone

Flavonoid

Flavonoid

Flavonoid

PE/ EtOAc 1.2:1

PE/ EtOAc 1.2:1

PE/ EtOAc 1.2:1

PE/ EtOAc 1.2:1

CCl4/EtOH 1.8:1

CCl4/EtOH 1.8:1

CCl4/EtOH 1.8:1

0.17

0.25

0.42

0.54

0.15

0.45

0.64

1.02

2.0

16.9

15.75

<1.5

3

10

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

O. gratissimum L.

Total alkaloids

Alkaloid

Alkaloid

Alkaloid

Terpene

Terpene

Terpene

Terpene

Terpene

Terpene

Terpene

-

EtOAc / n- hexane 1:1



EtOAc/PE 4:1

EtOAc/PE 4:1

EtOAc/PE 4:1

EtOAc/PE 4:1

EtOAc/PE 4:1

EtOAc/PE 4:1

EtOAc/PE 4:1

-

0.20

0.70

0.90

0.06

0.14

0.21

0.37

0.47

0.59

0.87

190

<65

<75

250

0.32

0.27

1.41

3.96

0.44

0.65

0.52

P. niruri L.

P. niruri L.

Total alkaloid

Alkaloid

-


-

0.23

100

<8 EtOAc: Ethyl Acetate PE: Petroleum Ether EtOH: Ethanol

Alkaloids: Alkaloids are one of the major

classes of compounds possessing Anti-malarial

activity (Ancolo et al., 2002; Kenny-Ang et al.,

2000; Bringmann et al., 2000). One of the

oldest and most important Anti-malarial drugs,

Quinine belongs to this class of compounds and

is still a very relevant therapy in the fight

against malaria. A number of naturally

occurring Alkaloids are reported to possess

Anti-malarial activity against different malarial

models. Crude Alkaloid extracts of C. papaya

Linn., O. gratissimum L. and P. nururi L. were

found to inhibit the growth of P. falciparum

with IC50 of 185µg/ml, 190µg/ml and

100µg/ml, respectively. The highest IC50 for

alkaloids (8µg/ml) was obtained for the

purified extract of P. nururi L. with an RF 0.23

(Ethyl acetate/Petroleum Ether, 4:1).

Terpenes: Artemisinin which is isolated from

Artemisia annua, a sesquiterpene lactone

endoperoxide, has been shown to possess

strong Anti-malarial activity. It has prompted

the investigation of some other naturally

occurring terpenoids for their schizonticidal

activity. Many were shown to affect the growth



of P. falciparum (Lin et al., 1987; Chukwejeku

et al., 2005; Uys et al., 2002). Ethanolic extract

of O. gratissimum L. revealed seven terpenes,

three of which possessed strong Anti-malarial

activity with RF of 0.06, 0.14, 0.47 and IC50 in

µg/ml of 0.32, 0.27 and 0.44 respectively. The

Ocimum species, commonly used in traditional

medicine in many parts of the world contain

Eugenol, Thymol and Geraniol as major

volatile oil constituents, whereas the major

flavones are Xanthomicrol and Cirsimaritin

(Vieira R.F. et al 2001).

Quinones: The structure of many naturally

occurring Quinones is based on Benzoquinone,

Naphtoquinone and Anthraquinone ring

systems. Naphtoquinone has been highly active

against P. falciparum in-vitro (Kapadia et al.,

2001). Ethanolic extract of G. kola Heckel.,

furnished four Quinones which were separated

on Silica gel plates with RF of 0.17, 0.25, 0.42

and 0.54 and values of IC50 in µg/ml of 1.02,

2.0, 16.9 and 15.75 respectively. It will be of

great interest to determine structure of at least

the Quinone with the lowest IC50 value.

Flavonoids: The Anti-malarial activity from

this class of compounds is not very common.

Nevertheless, Brandao et al. report that the

presence of Flavonoids in Bidens pilosa

explains its Anti-malarial activity (Brandao et

al., 2004). Ethanolic extract of G. kola Heckel.

gave three Flavonoids by preparative TLC

which showed remarkable inhibition on the

growth of P. falciparum with IC50 in µg/ml of

1.5, 3 and 10 respectively for RF of 0.15, 0.45

and 0.64.

Table 3: Traditional healer decoctions preparation and administered dosages

Plants Part Quantity used

for decoction;

30min of

boiling

Daily dosage

for adults

Treatment

duration

(days)

C. citratus

C. occidentalis

C. papaya Linn.

G. kola Heckel.

L. camara L.

O. gratissimum.

P. niruri L.

V. amygdalina

Leaves

Aerial part without seed

Leaves

Seeds

Leaves

Leaves

Aerial part

Leaves

10g in 1 l

100g in 1 l

200g in 1l

100g in 1 l

250g in 1 l

100g in 1 l

100g in 1 l

1×2C

1×1C

2×1C

2×1S to chew

1×1C

1×2C

1×1C

2×1C

4

5

5

5

5

6

5

5 C = 145ml; 1× or 2× = one or two times a day S = Seed

CONCLUSION

In the present study we showed that eight

remedies used by traditional healers to treat

malaria possess significant Anti-plasmodial

activity that was confirmed by inhibition of P.

falciparum growth in-vitro of crude alcoholic

extracts of these remedies. Some purified



extracts of Alkaloids, Terpenes, Quinones and

Flavonoids of the remedies are shown to inhibit

P. falciparum growth in-vitro and should be

responsible for the observed Anti-malarial

activity. It is necessary to determine structures

of these constituents in order to determine

efficiently in-vivo therapeutic dose for human

therapy. We will be working with companies

such as Cambridge Major Laboratories

(Germantown, WI, USA) on full structural

elucidation of the active constituents, and

possible synthesis of the active ingredients for

further testing.

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Source of Support: Nil Conflict of Interest: None Declared




PHARMACOGNOSTICAL AND PHYTOCHEMICAL EVALUATION OF

VIGNA UNGUICULATA LINN. (KULATTHA) SEED

Kolhe Rasika1, Acharya Rabinarayan

2 , Bhide Bhargav

3, Harisha CR

4, Shukla VJ

5

1 PG Scholar, Dravyaguna Department, IPGT&RA Gujarat Ayurved University, Jamnagar.

2 Associate Professor, Dravyaguna Department, IPGT&RA Gujarat Ayurved University, Jamnagar.

3 PhD scholar Dravyaguna Department, IPGT&RA Gujarat Ayurved University, Jamnagar.

4 Head, Pharmacognosy laboratory, IPGT&RA Gujarat Ayurved University, Jamnagar.

5 Head, Pharmaceutics laboratory, IPGT&RA Gujarat Ayurved University, Jamnagar.

*Corresponding Author - email - [email protected] mob.no.09374333651

Received: 03/02/2012; Revised: 17/02/12; Accepted: 29/02/12;

ABSTRACT

Kulattha (Vigna unguiculata Linn, Papilionaceae), one of the seed drugs described under dietetic

group, is being used as both drug and diet, in different classical texts of Ayurveda. It is one of the

drugs of choice for the management of urinary calculus (Ashmari). Though used as a source of both

drug and diet, it is reported as a major causative factor of acid peptic disorder (Amlapitta). Seed of

V.unguiculata can be identified microscopically by the presence of rhomboidal crystals, simple

starch grains with hilum. Purity test shows loss on drying (91.89% w/w), total ash (4.89% w/w), acid

insoluble ash (1.22% w/w), alcohol soluble extractive (1.31% w/w) and Water-soluble extractive

(1.94% w/w). Preliminary analysis revealed the presence of starch, tannin and amino acid. HPTLC

study of its methanolic extract showed the presence of four and seven spots in short and long UV

respectively. The information generated by this study provides relevant Pharmacognostical and

Physico-chemical data needed for proper identification and authentication of the seeds of this

particular species.

Key words: Kulattha, Pharmacognosy, Phyto-chemistry, HPTLC

Abbreviations: µm - micrometer



INTRODUCTION

Kulattha (Vigna unguiculata Linn.

Papilionaceae) is found throughout India from

Punjab and Himalaya to Sikkim in the North

(where it ascends to over 1500 m), the upper

Gangetic plain, Central and South India till

Kanyakumari. (Lakshmi1996). It is known as

Horse gram In English, Kulathi in Hindi,

Mudiraa in Malayalam, Kalathi in Gujarati,

Kulitha in Marathi. (Anonymus, Database on

Medicinal Plants 2008). It is extensively

cultivated in Dehradun, Bengal, Chotanagpur,

Deccan, and Coromandel Coast of Kerala as a

food crop especially for Horses and Cattles as

well as for its seeds which are considered

nutritious. (Lakshmi, 1996) Classical text of

Ayurveda, Charaka Samhita described it as one

of the drug/diet causing Acid Peptic Disorders

(Amlapitta) (Charaka Sutrasthana 25/40).

Sushruta Samhita highlights its Anti-lithiatic

(Shukrashmari) activity. (Sushruta Sutrasthana

46/37) In recent study V.unguiculata shows to

have better results than the use of conventional

potassium citrate in recurrence of renal calculus

and can be used to reduce the recurrence of

calcium oxalate stone. (Singh et al. 2010) Seed’s

extract also shows Anti-oxidant and Anti-free

radical activities. (Hazra B et al. 2009)

Plant morphology

It is a Sub-erect downy to rarely glabrescent

annual herb growing upto 30–60cm or more in

height with a short erect stem and several

elongate, diffuse, Sub-erect or at times twining

branches. The young shoot usually covered

with epidermal hairs, bearing pinnately

trifoliate alternate stipulate leaves having ovate

oblong or ovate lanceolate entire membraneous

stipellate leaflets 2.5 cm or more long, very

small pedicelled pale yellowish racemes

axillary, with 2–6 flowers clustered at top of

rachis and compressed, linear, falcate to much

curved, four to six seeded pods 3.85 cm long

and 6–9 mm wide (J.S. Gamble 1997).

Fruit: 4–6 seeded, with trichomes or downy

linear, broadly linear falcate or scimitar shaped

to much recurved (very slightly so in wild

variety) compressed pod. Measuring 3.8–5cm

long and 6–9mm broad tipped with the

persistent style. Seeds reniform, compressed

with a shiny hard testa of various colours,

mostly reddish brown, grey black as well as

mottled. Cotyledon orbicular to cordate and

persists for a long time on seedling.

Though used extensively as a drug and diet, the

detailed pharmacognostical and preliminary

phytochemical characters of the seeds are not

reported anywhere. Hence, in this article an

attempt has been made to study the

morphological and microscopical characters of

the seed and its powder along with preliminary

phytochemical characters including HPTLC

study.

MATERIAL AND METHODS

Kulattha seeds were purchased from the local

market in the month of March, authenticated by

Pharmacognosy laboratory of IPGT&RA

Jamnagar. Voucher herbarium specimen along

with crude drug sample was preserved in

Pharmacognosy laboratory, vide Ref No. 6036

March 2011. Pharmacognostical evaluation of

Kulattha seeds including Histo-chemical

studies were carried out by taking free hand

sections. Powder microscopy of seeds was

carried out following standard procedures

(Wallis 1985) and the slides for powder

microscopy were separately prepared first with

distilled water, then stained with

Phloroglucinol and concentrated HCl.

Photomicrographs were taken using Carl Zeiss

Binocular Microscope attached with Camera.

Histo-chemical tests were carried out by taking

thick sections following the standard procedure



methods (Krishnamurthy 1988). Physico-

chemical parameters and chemical screening

(Anonymous, Planner Chromatography 1999)

studies were carried out following standard

procedures. (Anonymous, API 2004) The

methanol extract obtained during Physico-

chemical parameters was used for HPTLC.

Sample was prepared by 30min sonication of

drugs with methanolic medium and filtrate was

used for experimental task. ‘Benzene’ was

selected as the mobile phase. Chromatographic

conditions were as follows. (Anonymous,

Planner Chromatography 1999)

Chromatographic conditions

Application mode : Camag Linomat V

Development Chamber : Camag Twin trough Chamber.

Plates : Precoated Silica Gel GF254 Plates.

Chamber Saturation : 30 min.

Development Time : 30 min.

Development distance : 7 cm.

Scanner : Camag Scanner III.

Detection : Deuterium lamp, Tungsten lamp

Data System : Win cats software.

RESULTS AND DISCUSION

Macroscopic Characters of seed

Seeds shape reniform, 5–6mm long, 3–4mm

broad and 2–3 mm in thickness, compressed

with a polished or shiny and hard brown

coloured testa. The micropyle was situated near

the hilum. The hilum was 1–1.5 mm in length.

The seed were exalbuminous. The testa was

tough but comparatively thin except at the

region of the hilum. The embryo which was

exposed after removing the testa, by softening

it through emersion of the seed in water,

consists of two fleshy cotyledons, 5–6mm long

and 4–5mm wide and an incurved radical

which was 4mm long. [Figure 1.1]

Organoleptic evaluation

Organoleptic evaluation of powder of seeds of

V. unguiculata revealed astringent (kashaya)

taste, buff colour and characteristic odour while

the texture was coarse. [Figure 1.2]

Microscopic Character:

T.S. of Seed

Detailed Transverse section passing through the

center of the seed shows the testa having three

layers, of which first and second layer were

single celled, while the third layer consisted of

several rows of thin walled narrow cells

containing rhomboidal calcium oxalate crystals.

The outermost row namely the epidermis was

composed of vertically elongated palisade like

cells, each cell with slight constriction nearer

its upper end, 45µm or more in height, 12–

15µm in width at their broader part, 3–6µm

width at their narrow constricted part and 9–

12µm in width at the extreme tip. [Figure 1.3,

1.4] There was a thin covering of cuticle over

the epidermis about 3µm in thickness. The

palisade like cells were further characterized by

the presence of narrow transverse light line at

about two-third of their length from the base.

The second or Sub-epidermal row consisted of

shorter and broader column like cells with their

outer and inner ends broader and the middle



portion constricted. Cells with inter-cellular

spaces had characteristic “hour glass” like

shapes. They varied from 18–30µm in height

and 12–15µm in width at the base, 6–9µm in

width at the narrow middle constricted region

and 15–18µm near the top. [Figure 1.5]. Third

layer or zone was composed of 8–10 rows of

thin walled narrow cells, some of which

contained rhomboidal crystals that measured

30–60µm × 18–27µm, these parenchymatous

cells were tangentially elongated with more or

less oblique radical walls. These cells were

lacking the inter cellular spaces. At the region

of the hilum, two rows of palisade like cells

were present (instead of one row). Beneath,

there was a group of sclerenchyma cells with

narrow elongated pits on their walls that

appeared as an elongate lanceolate patch in T.S.

The Sub-epidermal cells were columnar,

expanded beneath the hilum into a cushion in

which these groups of sclerenchymatous cells

appeared embedded, surrounded by two layers

of narrow thin walled elongated

parenchymatous cells. [Figure1. 6, 1.7]

Plate 1: Microscopic characters of V.unguiculata

Fig.1.1 Seeds of V.unguiculata

Fig. 1.2 Seed Powder of

V.unguiculata

Fig. 1.3 Rhombidal shaped

calcium oxalate crystal

Fig. 1.4 T.S of V.unguiculata

seed with testa

Fig. 1.5 T.S. of testa

+cotyledon

Fig. 1.6 T.S. showing

cotyledon



Fig. 1.7 T.S. of cotyledon

stain with iodine

Fig. 1.8 Protein content

Fig. 1.9 Lignified fiber

Fig. 1.10 Starch with tannin

material

Fig. 1.11 Simple starch

grain with hilum

Fig. 1.12 Prismatic

Crystals

Powder Microscopy

Diagnostic characters of dried powder of V.

unguiculata under the Microscope were

prismatic crystals of Calcium oxalate from

epidermis, dark brown coloured content; which

was confirmed to be Tannin by adding Ferric

chloride solution to it, which turned black;

from Sub-epidermal region, simple starch

grains with Hilum, Iodine stained starch grains,

Loosely arranged parenchymatous cells.

Clumped masses may be protein content, with

some Aleurone grains. [Figure 1.8 to1.12]

Histochemical Test:

The results of the various Histo-chemical tests

carried out, to detect Lignin, Calcium, Starch

and Tannin are depicted in table no. 1

Table -1 Histochemical evaluation of sections of V. unguiculata seed.

Material Reagent Test for Result

Section Phloroglucinol+Conc

HCl

Lignin Present

Section Phloroglucinol+Conc

HCl

Calcium oxalate

crystals

Present

Section KI Starch Present

Section Ferric chloride

solution

Tannin Present

Section Conc.H2SO4 Protein Present



Phyto-chemical constituents

While observing the physicochemical

characters for purity test, the loss on drying was

not more than 91.89% w/w, ash value was not

more than 4.89 % w/w, acid insoluble ash was

not more than 58.14% w/w, water soluble

extractive was not more than 1.94 % w/w and

the methanol soluble extractive was not more

than 1.34 % w/w. [Table no.2] Qualitative

analysis showed presence of tannin, starch,

amino acid and absence of alkaloids,

cyanogenic glycosides and flavonoids. [Table

no.3]. The methanol soluble extract was

examined for high performance thin layer

chromatography profile at 254nm frequency,

using the solvent system of Benzen, Under

high performance thin layer chromatography

profile, at 254nm frequency four peaks were

observed and at 366nm frequency seven peaks

were observed. [Table no.4] [Plate 2]

Table-2 Showing the physicochemical parameters of V. unguiculata

Parameter Observation

Loss on drying 91.89

Ash value (%w/w) 4.89

Acid insoluble ash (%w/w) 1.22

Water soluble extract (%w/w) 1.94

Alcohol soluble extract (%w/w) 1.31

Table- 3 Showing the results of qualitative test for various functional groups in V. unguiculata

Active constituents Result

Tannin Present

Flavonoids Absent

Starch Present

Alkaloid Absent

Amino acid Present

Cyanogenetic glycoside Absent

Table- 4 HPTLC Profile of V. unguiculata

No. of spots Rf

Short UV 254 4 0.02,.0.01,0.36,0.44

Long UV 366 7 0.04,0.09,0.14,0.18,0.32,0.46,0.55



Plate 2 : Methanol soluble extractive Chromatogram of V.unguiculata

254 nm

366 nm

After spray

CONCLUSION

Pharmacognostical and Phytochemical

evaluation of seed and powder of Kulattha

(V.unguiculata) market sample were found to

be authentic and meet the standard parameters

of API. Further Pharmacognostical findings i.e.

rhomboidal calcium oxalate crystals, tannin,

lignified fibres were important characters found

during the study apart from what was

mentioned in API. Phytochemical screening

and HPTLC results can be considered as

standards for further research works.

ACKNOWLEGMENTS

Author is thankful to the authorities of IPGT &

RA, Jamnagar for providing facilities to carry

out the research work.

REFERENCES

Anonymous, Ayurvedic Pharmacopoeia of

India (2004). Part I, Vol I, Appendix 2.1.4,

2.1.5 and 2.1.7 New Delhi: Government of

India publication;

Anonymous, Database on Medicinal Plants

(2008) , Vol 5, NewDelhi; CCRAS publication

; p.no.123

Anonymous, Planner Chromatography, Modern

Thin layer Chromatography, Switzerland

(1999), pg. 2–16

Anonymous, The Ayurvedic Pharmacopoeia of

India (2004) Part I, Vol-III, Appendix 2.2.3,

New Delhi: Government of India

Publication; p.no.234

Gupta Ram Bhagawat (2003) Ayurveda Ka

Pramanika Itihas, Chaukhamba Krishnadas

Academy, Varanasi, p.no. 265

Gupta Ram Bhagawat, (2003) Ayurveda Ka

Pramanika Itihas, Chaukhamba Krishnadas

Academy, Varanasi, p.no.247

Hazra B, Sarkar R, Mandal S, Biswas S, and

Mandal N, (2009). Studies on antioxidant and

antiradical activities of Dolichos biflorus seed

extract, African Journal of Biotechnology. 8

(16): 3682–398

J.S. Gamble, Flora of the

Presidency of Madras, Vol.1, 14 old Connaught

place, Dehraduna, India; p.no.366



Kaviraj Kunjalal Bhishagratna,(2005) Sushruta

Samhita, English translation, Vol I,

Chaukhamba Sanskrit series office, Varanasi,

Sutrasthana 46/36, p.no.455

Krishnamurty, K.V.(1988), Methods in the

plant histochemistry, Vishwanadhan Pvt,

Limited, Madras, p.no.1–77.

Lakshmi N (1996) Pharmacognosy of

Ayurvedic drugs Kerala, Pharmacognosy Unit,

Ayurveda research institute,

Poojapura,Thiruvananthapuram. p.no.31

Lakshmi N (1996) Pharmacognosy of

Ayurvedic drugs Kerala, Pharmacognosy Unit,

Ayurveda research institute,

Poojapura,Thiruvananthapuram. p.no.32

Rana Gopal Singh, Sanjeev Kumar Behura,

Rakesh Kumar (2010), Litholytic Property of

Kulattha (Dolichous Biflorus) vs Potassium

Citrate in Renal Calculus Disease: A

Comparative Study, JAPI May, Vol.58

Sharma P V,(2009) Charaka Samhita, English

translation, Vol I, Chaukhamba oriantalia,

Sutrasthana 25/40; p.no.169

Wallis TE. (1985) Textbook of

Pharmacognosy, London: Churchill Publication

; p.no.572–82





RUBIA CORDIFOLIA LINN. (MANJISHTHA) IN PRIMARY

DYSMENORRHOEA (KASHTARTAVA) - A CLINICAL STUDY

Dhiman Kamini1*, Lata Kusum

2, Dhiman K. S

3.

1

Department of Stri Roga & Prasooti Tantra, IPGT & RA, Gujarat Ayurved University, Jamnagar 2

Department of Stri Roga & Prasooti Tantra, Ayurvedic College, Hoshiarpur 3 Prof. & Head, Deparment of Shalakya, IPGT & RA, Gujarat Ayurved University, Jamnagar,

Gujarat, India

*Corresponding Author: [email protected]

Received: 09/02/2012; Revised: 21/02/2012; Accepted: 05/03/2012;

ABSTRACT

Dysmenorrhoea is one of the most common gynaecologic complaints in young women who present

to clinicians now a days. There are references that Rubia cordifolia L. (Manjishtha) was in use

traditionally for Dysmenorrhoea in India and China since ancient times. On this basis a clinical study

to evaluate its efficacy in Dysmenorrhoea (Kashtartava) was undertaken with a total number of 20

patients having been registered for the study. Manjistha Churna orally with luke warm water in a

dose of 3 gms twice a day(12 hrly) was given for the duration of 2 months. Statistically highly

significant results were observed in the study.

Key Words: Dysmenorrhoea, Kashtartava, Artavadushti, Manjishtha, Rubia Cordifolia L,

Raktaprasadana



INTRODUCTION

Menstruation is a normal physiological

process indicating womanhood. It is a cyclical

process which repeats every month and

becomes an important physiological

manifestation in a woman’s reproductive life

Menstruation normally flows without pain and

burning sensation and the flow is not unctuous.

(Charak 700 BC)

When menstruation is

associated with pain, it is termed as

Dysmenorrhoea; the pain incapacitates day-to-

day activities of a female (Dutta D. C. 2007).

Dysmenorrhoea is a major cause of

absenteeism from work amongst women thus

decreasing efficiency and quality of life among

affected Women (Balbi et al., 2009). When

painful menstruation is present in Women with

normal pelvic anatomy, is defined as Primary

Dysmenorrhoea, usually begins during

adolescence (Wentz Anne Colston1985).

Affected women experience sharp, intermittent

spasm of pain usually concentrated in the supra

pubic area. Pain may radiate to back of the legs

(Thigh region) or the lower back. Systemic

symptoms include Nausea, Vomiting,

Diarrhoea, Fatigue, mild Fever and Headache

or Light headedness (Dutta D. C. 2007).

Painful menstruation with pelvic pathology is

defined as Secondary Dysmenorrhoea (Shah

PK et al., 2011).

Aartava and Menstruation both convey

the same meaning. ‘Kashta’ means painful,

difficult & ‘Aartava’ means menstruation.

Hence, Kashtartava refers to “Kashtena

munchyati iti Kashtartava” i.e. the condition

where in Aartava is shed with pain is termed as

Kashtartava (Dysmenorrhoea). In Ayurvedic

texts, though Kashtartava (Dysmenorrhoea) is

not separately described as a disease but there

are many diseases in which Kashtartava has

been mentioned as a symptom.

Dysmenorrhoea is one of the most common

gynaecological complaints in young women

who present to clinicians now a days. Modern

life style changes, faulty dietary habits and

stress seem to be few important causes for

Dysmenorrhoea. In this today’s high-tech era,

where the females are in par with male in

getting education or in their profession, don’t

take chance of skipping their duty in every

cycle and also reduced pain threshold in

females is one of the reason for increased

reporting of incidence of dysmenorrhoea.

Analgesics and NSAIDS which are active

inhibitors of PGs synthesis are used to combat

with pain during Dysmenorrhoea but these

drugs produce side effects on long term use

(Campbell, et al., 1997). Hence it is a need of

the day to find safe and effective remedy for

this problem.

It is a well-known fact that Ayurvedic

System of medicine always played an

important role in meeting the global health care

needs which is true even in the case of

management of dysmenorrhoea. In recent

times, focus on plant research has got increased

all over the world.

Rubia cordifolia Linn. (Rubiaceae),

popularly known as Indian Madder or majit or

manjishtha is also one of the important drugs

which is time tested. It is a perennial,

herbaceous climber with very long, cylindrical

roots with a thin red bark (Kirtikar & Basu

1980). Stems are long, rough, grooved and

become slightly woody at the base. It is found

throughout the hilly districts of India from

North-west Himalayas Eastwards, ascending to

8000ft. and Southwards to Ceylon. Generally,

root, leaves, fruits, stem etc. of the plant Rubia

cordifolia are used for their therapeutic

properties. The roots of this plant are of high

medicinal value and are traditionally used as

Anti-inflammatory, Astringent, Tonic, Anti-

septic, Diuretic, Anti-dysenteric, Blood

purifier, Anti-helminthic, Analgesic,

Hepatoprotective etc [(Kirtikar & Basu (1980),

Williamson Elizabeth (2002), Khare (2004),

Anonymous (2005)]. There are references that

Rubia Cordifolia (Manjishtha) was in use

traditionally for Dysmenorrhoea in India and

China [Guangzhou (1992) Hocking (1997)].

But till date there are no scientific references

available of any study using Manjishta as a

single drug in Dysmenorrhoea, keeping this

fact in mind present clinical study had been

undertaken to evaluate and establish its efficacy

in Dysmennorhoea (Kashtartava).



MATERIAL AND METHODS

Materials: Here, for the clinical study dried

root of Manjishtha(Rubia cordifolia L.) were

collected and identified in the Dravya Guna

Department, and then fine powder was done in

the Pharmacy of RGGPG Ayurved College,

Paprola (HP).

Methods:

Clinical Study: Patients attending the OPD of

Prasooti Tantra and Stree roga at Rajiv Gandhi

Govt. Ayurvedic Hospital, attached with Rajiv

Gandhi Govt. Post Graduate Ayurvedic

College, Paprola Distt Kangra (H.P.) with

characteristic features of Kashtartava

(Dysmenorrhoea) were selected for the present

study. The patients were selected and registered

irrespective of their caste, creed, religion,

income, occupation, etc.

Study Design: Single blind, prospective observational study.

Plan of the Study

Inclusion Criteria:

1. Patients of the age group of 10−50 yrs

2. Patients presenting sign and symptoms of Kashtartava(Dysmenorrhoea)

Criteria for Exclusion:

1. Patients not willing for trial

2. Patients having irregular periods

3. Patients having heavy and excessive periods

4. Patients having any anatomical anomaly of Reproductive System

5. Patients suffering from any systemic disease

6. Patients having conditions where surgical intervention was needed

Method of Study: - This Clinical study was

accomplished in three phases:

i) Diagnostic Phase, ii) Interventional Phase

and iii) Assessment Phase

Diagnostic Phase: Total 20 patients were

diagnosed on the basis of signs and symptoms

(Clinical presentation) of Dysmenorrhoea

(Kashtartava)and selected for the study after

following inclusion and exclusion criteria. The

nature of the study was explained to all the

selected patients and their consent (informed

consent) was obtained. A special clinical

Proforma was prepared incorporating both

Ayurvedic (Dashavidha Pareeksha) and

Modern parameters. A detailed history was

taken and complete physical examination and

laboratory investigations were also carried out.

Subjective Criteria adopted for the present

study were –

� Intensity of Pain

� Duration of Pain

� Nausea

� Vomiting

� Breast Tenderness

� Fever

� Headache

� Vertigo

� Diarrhoea

� Anorexia & Nervousness

Objective Parameters: For the purpose of

assessing the general condition of the patient

and to exclude the other pathologies routine

Haematological and USG (Abdomen & Pelvis)

examination was carried out. After arriving at

the diagnosis, clinical Proforma was filled up,

which incorporated all the signs and symptoms

based on both Ayurvedic as well as Modern

Parameters. A detailed clinical history was

taken initially and complete physical and

Gynaecological examination of each patient



was carried out and the same was recorded in

the proforma.

Interventional Phase: The study was

intervened by the treatment with Powder of

Rubia Cordifolia L.( Manjistha Churna).

Drug Schedule: Manjistha Churna (Powder of

Rubia cordifolia L) orally with luke warm

water in a dose of 3gms twice a day (12 hrly)

was given for a duration of 2 months.

Duration of Trial: The total duration of

treatment for the subjects was two months.

Follow Up Study: Follow up was conducted

after one month during trial and then after the

completion of trial.

Assessment Phase: The effect of treatment

(results) was assessed regarding the clinical

signs and symptoms (on the basis of VAS and

grading, scoring system) and the overall

improvement was observed and recorded as

Before Treatment (BT) and After Treatment

(AT).

Clinical Assessment: The criteria adopted for

intensity of pain was VAS (Visual Analogue

Scale). Visual Analogue Scale (VAS) is a

measurement instrument that tries to measure a

characteristic or attitude that is believed to

range across a continuum of values and which

cannot be easily and directly measured.

Operationally a VAS is usually a

horizontal line, 100 mm (10 cm) in length,

anchored by word descriptor at each end, as

illustrated in figure-1.

The patient marks on the line the point that they

feel represent their perception of their current

state. The VAS score is determined by

measuring in millimetres from the left hand end

of the line to the point that the patient marks.

The signs and symptoms were assessed by

adopting suitable scoring methods. The details

are illustrated in Table 1.

Figure – 1, Visual Analogue Scale (VAS)



Table -1 Showing grading of Pain in Patients

Symptoms 0 1 2 3

1. Duration of pain No pain Upto 24 hr. Upto 48 hr. Upto 72 hr.

2. Nausea Absent 2–3

times/day 4–5 times/day >5 times/day

3.Vomiting Absent Occasionally 1–2 times/day >2 times/day

4.Breast tenderness

No

Tenderness

Mild

Tenderness

Moderate

tenderness

Severe

Tenderness

5. Fever

No fever

Mild fever at

night

Moderate fever

throughout the

day

Severe fever

6. Headache Absent Mild Moderate Severe

7. Vertigo

Absent

Occasionally

2–3 times in 1–

2 days

More than 4

times in 3–4

days

8. Diarrhoea Absent Occasionally 2–3 times/day >3 times/day

9. Anorexia Absent Mild Moderate Severe

10. Nervousness Absent Mild Moderate Severe

Statistical Analysis: The obtained data were

analyzed statistically and presented as Standard

Error of Mean. The observed difference was

calculated by adopting student’s “t” test

(Significance level: ≥0.05).

Overall Assessment of Therapy: To assess

the overall effect of therapy following criteria

was laid down.

Completely Cured: More than 90% relief in

symptoms and signs as well as one (1) score

obtained according to VAS.

Markedly Improved: More than 75% and less

than 90% relief in symptoms and signs as well

as > 1–5 change of score are according to VAS.

Moderately Improved: More than 50% and

less than 75% relief in symptoms and signs as

well as 5–9 change of score according to VAS.

No improvement/ Unchanged: Less than 25%

relief in signs and symptoms and score greater

than 9. The total effect of therapy was assessed

on the basis of the above first three criteria.

RESULTS

Total 20 patients were registered, among them

65.00% patients belonged to age group of 18–

25 years and 70% were unmarried. Most of the

patients (70.00%) were students and maximum

(75.00%) were of lower middle class. All the

patients were of Hindu religion and from rural

area. Maximum patients(85.00%) were

vegetarian; 80.00% patients had spicy food as

their dietary habits and 20.00% were on non

spicy diet. 45.00% patients were used to

consume Amla rasa and 40.00% were used to

consume Lavana rasa dominantly. 85.00%

patients had disturbed sleep due to pain and

15.00% had normal sleep pattern. Maximum

number (90.00%) of patients had Menarche

between 13–15 years. 40% patients had 4–5

days, 35.00% and 25.00% had bleeding P/V for

3 days & more than 5 days respectively.

85.00% patients had interval of 26–30 days,

15.00% patients had 31–35 days interval in

between two Menstrual cycles. The amount of

blood loss was scanty (75.00%) in maximum

number of patients & moderate flow in rest

(25.00%) of the patients. Maximum number of

patients (07.00%) were having spasmodic pain

and 25.00% were having dull ache. Maximum

number of patients had radiation of pain to



thighs (50.00 %), to abdomen (45.00%) and

05.00% in the back. 60.00% patients were

having Nausea, 30.00% patients had Anorexia,

25.00% were having Fever, 10.00% had

Headache, and 05.00% had vertigo as

associated Symptoms. Majority were of Vata

Pittaja (85.00%), Pitta Kaphaja (10.00%) &

Vata kaphaja prakriti (05.00%).

Table -2 STATISTICAL ANALYSIS OF EFFECT OF THERAPY

RUBIA CORDIFOLIA L. (MANJISTHA CHURNA)

Symptom N Mean score Relief SD+ SE+ t p

B.T. A.T. Diff. In %

Intensity of pain 20 8.97 1.99 6.98 77.82 0.597 0.133 52.48 <0.001

Duration of pain 20 1.3 0.25 1.05 80.77 0.2236 0.499 21.00 <0.001

Associated

Symptoms

Nausea 20 0.7 0 0.7 100 0.6569 0.1468 4.768 <0.001

Vomiting 20 - - - - - - - -

Tenderness 20 0.05 0 0.05 100 0.2236 0.05 1 >0.05

Fever 20 0.25 0 0.25 100 0.4442 0.0993 2.51 <0.05

Headache 20 0.1 0 0.1 100 0.3077 0.06882 1.4530 >0.05

Vertigo 20 0.05 0 0.05 100 0.2236 0.05 1 >0.05

Diarrhoea 20 0.05 0 0.05 100 0.2236 0.05 1 >0.05

Anorexia 20 0.3 0 0.3 100 0.4701 0.105 2.857 =0.01

Nervousness 20 - - - - - - - -

Intensity of pain: The initial mean score of

intensity of pain was 8.97 before the treatment

which was reduced to 1.99 after the treatment.

The percentage of relief was 77.82% which is

significant statistically at the level of p<0.001

(t=52.48).

Duration of pain: The initial mean score of

duration of pain was 1.3 before the treatment

which was reduced to 0.25 after the treatment.

The percentage of relief was 80.77% which is

significant statistically at the level of p<0.001

(t=21.00)

Nausea: The initial mean score was 0.7 before

the treatment which was reduced to 0 after the

treatment. The percentage of relief was 100%

which is not significant statistically at the level

of p<0.001 (t = 4.768).

Vomiting: Was not reported in any patient.

Breast tenderness: The initial mean score was

0.05 before the treatment which was reduced to

0 after the treatment. The percentage of relief

was 100% which is not significant statistically

at the level of p>0.05 (t=1).

Fever: The initial mean score was 0.25 before

the treatment which was reduced to 0 after the

treatment. The percentage of relief was 100%

which is significant statistically at the level

of p<0.05 (t=2.51).

Headache: The initial mean score was 0.1

before the treatment which was reduced to 0

after the treatment. The percentage of relief


at the level of p>0.05 (t=1.4530).

Vertigo: The initial mean score was 0.05




at the level of p>0.05 (t=1).

Diarrhoea: The initial mean score was 0.05


after the treatment. The percent of relief was

100% which is not significant statistically at the

level of p >0.05 (t=1).

Anorexia: The initial mean score was 0.3



was 100%, which is significant statistically at

the level of p=0.01 (t=2.857).



Nervousness: Was not reported even in a

single patient.

Overall Results

The result was assessed on the basis of VAS

and Grading, Scoring system 14(70%) patients

were completely cured, 1(5%) patient markedly

improved & 5 (25%) were moderately

improved.

DISCUSSION

In Ayurvedic classic Kashtartava is not

considered as a separate disease however

mentioned as a symptom of various diseases

(Charak 700 BC) like Vatala, Sannipatika and

Udavarta Yonivyapad. Commentator

Chakrapani (Charak 700 BC) says that any

symptom may manifest as a separate disease

Acharya Sushruta while depicting the

importance of Shuddha Artava (Menstrual

blood) has assigned a separate chapter in

Sharira Sthana and has quoted Kashtartava as

a type of Artava Dushti (Menstrual Disorder).

According to Acharya Kashyapa, the blood in

adult females during their reproductive period

enters into Garbha Kosha (Uterus) every

month and Rajovaha Shiras (vessels carrying

menstrual blood) in the uterus carries the

Artava (Menstrual blood) formed by the action

of Artavagni upon the Rakta (Blood) and fill

the uterus in one month and after that this

Artava (menstrual blood) is expelled out by

these Shiras (blood vessels) at the interval of

one month (Charak 700BC).

In Sushruta Samhita, (Sushruta 600 BC)

characteristics of Shuddha artava (Menstrual

blood) is mentioned as

“Raktalakshanam”(having characteristics of

blood) and it prepares the Garbhashaya

(Uterus) to receive the fertilized egg as well as

for the growth and development of the foetus21

.

All these references reflect that Artava

(Menstrual blood) is having similar qualities of

Rakta (Blood) hence if Rakta (Blood) is pure

and having no any impurities there will not be

any disorder related to Artava (Menstruation)

but if Rakta vitiates there will be artavadushti

(Menstrual disorder) too which results in

Gynaecological disorders like Kashtartava

(Dysmenorrhoea).

From the observed results on signs and

symptoms, it can be revealed that Rubia

Cordifolia L.(Manjishtha), which is

Raktaprasadak dravya (Blood purifying agent)

eliminates and nullifies all types of impurity,

toxicity, contamination and harmful effect of

unwanted material from blood and restores its

health (Brahmashankara Mishra, 1993). As

maximum patients were having a tendency to

consume Amla (sour) and Lavana rasa (salty

taste) predominantly which are responsible to

cause Raktadushti (vitiation of blood).

Furthermore, Artavadushti (Menstrual disorder

(Charak 700 BC) itself manifests as

Kashtartava (Dysmenorrhoea) and to alleviate

dushti (impurities), prasadana (purification) is

much supportive.

Raktavardhaka (Blood enhancing) property of

Manjishtha (Rubia Cordifolia L.) improves

pain threshold by improving the Immunity. The

biological investigations have shown that many

of the medicinal properties claimed for Rubia

Cordifolia (Manjishtha) in the historical texts

have sound scientific basis (Singh et al., 2005).

It has a variety of uses such as blood purifying

etc. (Joharapurkar et al., 2003). Intensity and

duration of pain was observed to be reduced in

maximum patients. Anti-inflammatory and

Analgesic activity of Rubia cordifolia is quoted

by many Researchers. [Mhaskar, Blatter &

Caius (2000), Mooradian (1988), Kasture,

Deshmukh(2000), Khare (2007)].

Rubia cordifolia L. is said to contain

substantial amount of Anthraquinones,

especially in the roots which is responsible for

its pharmacological activities (e.g. Anti-

inflammatory and Analgesic). (Meena et al.,

2010); (Anar Patel et al., 2010); (Yeungnam

2007), (Gonzalez, et al., 1974); (Schildknecht

et al., 1976); (Itokawa et al., 1989); (Ho, et al.,

1996); (Hua, et al., 1992); (Kawasaki et al.,

1992); (Marec et al., 2001); (Chung et al.,

1994); (Antarkar et al., 1983); (Tripathi and

Sharma 1998); (Wealth of India 2002).

Psychological factors like Depression, Anxiety

and Stress are quoted as the risk factors of

Dysmenorrhoea (L Wang et al., 2004) and due

to Anti-stress property (Patil, Jagdale et al.,



2006) of Rubia cordifolia (Manjishtha) it is

more likely to modify the factor like pain in

Kashtartava (Dysmenorrhoea).

Prostaglandins and Prostinoids are

biosynthesized from Arachidonic acid through

the COX pathway after production of

Arachidonic acid from hydrolysis of

Phospholipids by Phospholipase. In excess or

imbalanced amount of Prostinoids released

from the Endometrium during menstruation

induces uterus to contract frequently and

dysrhythmically, with increased basal tone and

increased active pressure. Uterine hyper

contractility, reduced uterine blood flow and

increased peripheral nerve hypersensitivity

induce pain. Mollugin is one of the major 2H

naphtho pyran component isolated from Rubia

cordifolia is having strong inhibitory activity

on arachidonic acid. Thus Rubia cordifolia

L.(Manjishtha) by breaking the pathway of

pain production improves the condition like

Dysmenorrhoea.

CONCLUSION

Analysis of the data of the present study

revealed Manjishtha (Rubia Cordifolia L.)

churna has significant role in the management

of Kashtartava (Dysmenorrhoea). Though the

results are good, but further study on large

numbers of patients should be done along with

some specific investigations like Prostaglandin

synthetase evaluation.

ACKNOWLEDGEMENTS

The author thank the staff of labs., hospital,

Pharmacy of the College for their help and

support of patients for participating in this

study.

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