2. REVIEW OF LITERATURE

2.1. GENERAL

Pharmacognosy may be defined as “an applied science that deals

with the biologic, biochemical and economic feature of natural drugs and

their constituents.” Modern aspects of science include not only the crude

drugs but also their natural derivatives (Tyler et al., 1981).

Shah and Qadry (2005) defined pharmacognosy as the study of

crude drugs obtained from plants, animals and mineral kingdom and their

constituents. It means knowledge or science of drugs. Most of the crude

drugs used in medicine are obtained from plants which after collection are

subjected only to drying or making them into transverse or longitudinal

slices or peeling them in some cases.

Morphological characters are used to identify the crude drugs,

whatever it may be entire plant, aerial parts, stem, leaves, flower, root etc.,

similarly, if the crude drug in broken stage or even in powdered condition,

through the Microscopical characters the arrangement of tissues are

identified.

The diagnostic elements such as stomata, trichomes, vessels, stone cells,

starch grains, calcium oxalate crystals persist in powdered condition and

are used in identification of drugs.

12

Linear measurements as diameter of starch grains or length and breadth of

fibres and vessels are also useful in identification of closely related

drugs/species.

Quantitative microscopic constants like vein islet number; palisade

ratio and stomatal index are also important tools or the same purpose.

If foreign organic or inorganic matter is excess than the

pharmacopoeial limits, the drug is not according to standard. For these

reasons, some of the preliminary parameters are needed for determining

the standard drug.

I. tinctoria is a shrub, leaves pinnately compound, leaflets 9-14. Flowers

pink, pod long linear 8-12 seeded, seeds brown and rare on wastelands.

Historical background, therapeutic activity, tinctorial importance,

cultivation of I. tinctoria, dye content, preparation method and chemical

composition of indigo dye have been discussed by Siddiqui and Khair,

1998.

Tephrosia is a genus which belongs to the family Fabaceae. The

genus name Tephrosia derives from the Greek word “tephr (o)” meaning

“ashes, ash colored, and gray” as the colour of the stems, leaves and fruit

of all the species are gray in colour.

13

More works have been done on some of the members of this family.

Botany, Taxonomic status, Pharmacognosy studies on

T. purpurea have been reviewed by Zafar and Mujeeb (2000).

The seedling morphology of T. purpurea, T. maxima, T. pumila and T.

villosa observed for 45 days and described. The intercalary meristems

were very active in T. villosa (internode to 5cm) and very minimum in T.

maxima (0.2cm) (Augustine, 2000).

2.2. SYSTEMATIC POSITION OF T. purpurea L.

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnoliopside

Order : Fabales

Family : Leguminosae (Fabaceae)

Genus : Tephrosia

Species : purpurea (L.)

2.2.1. Vernacular Names

Tamil : Kolingi

English : Purple Tephrosia

Sans. : Sharapunka

Tel. : Vempali

Mal. : Kozhenjil, Kattamiri

Beng. : Sarphonka

14

Guj. : Ghodakan

Ceylon : Kavilai

2.2.2. Distribution

Through the plains of India, Ceylon, Mauritius, Tropical Africa and

subtropical regions.

2.2.3. Description

Subshrub to 1mm; branchlets pubescent – villous. Leaves to 7 cm;

leaflets 4 -9 pairs, obovate, 0.8 – 2 × 0.3 – 0.7 cm, pubescent, base

cuneate, margin entire, apex obtuse, mucronate; petiole to 1 cm; petiolule

1mm; stipules, lanceolate, 5mm. Pseudoracemes leaf-opposed, to 8 cm;

bract to 2mm, pubescent; lobes lanceolate; upper lobes 2.5 mm, equal to

lower one. Corolla bluish-pink to purple; standard orbicular, 8.5 × 8mm,

sericeous; wings 7.5 × 3 mm; keels 6.5 × 2.5 mm; staminal sheath 5 mm;

filaments 2 mm. Ovary 5 mm, appressed – pubescent; style 3 mm,

glabrous. Pod 4 × 0.4 cm, downy – puberulous, continuous within,

slightly falcate; seeds ca. 7, ovoid, 3.5 mm, strophiole in the middle of

seed (Mathew, 1983).

2.2.4. Useful Part

Leaves, root, root bark and aerial parts.

15

2.2.5. Medicinal Uses

Tonic, laxative, diuretic and deobstruent; used in bronchitis and

bilious febrile attacks, and also for boils, pimples and bleeding piles.

Pharmacological studies have shown that extract of the herb is useful in

insufficiency of liver, but is not effective in infantile cirrhosis. Roots and

seeds are insecticidal and piscicidal (reports conflicting). Decoction of

roots given in dyspepsia, diarrhoea, rheumatism, asthma and urinary

disorders; roots given with black pepper in colic. A liniment prepared

from the roots is used in elephantiasis. Pulverized roots smoked for relief

from asthma and cough, decoction of pods used as a vermifuge and to stop

vomiting. Seeds yields oil said to be specific against scabies, itch, eczema

and other skin eruptions.

Fresh root-bark, ground and made into a pill, with a little black-

pepper, is frequently given in cases of obstinate colic. The plant is used

internally as a purifier of the blood, and is considered a cordiac tonic. An

infusion of the seeds is given as a cooling medicine.

In Ceylon, it is employed as an anthelmintic for children.

In French Guiana, the root is used to stupefy or fish poison.

This drug is said to be useful in cough and in derangement of the

kidneys. A decoction of the drug was administered in one ounce doses to

cases of Bright’s disease with dropsy and found to possess diuretic

properties in a mild degree (Kritikar and Basu, 1980).

16

2.3. SYSTEMATIC POSITION OF T. villosa Pers.

Kingdom : Plantae

Division : Magnoliophyta

Class : Magnoliopsida

Order : Fabales

Family : Leguminosae (Fabaceae)

Genus : Tephrosia

Species : villosa Pers.

2.3.1. Vernacular Names

Tam. : Vaykkaralai, Punaikkayvelai

Tel. : Nooguvempali

Guj. : Runchhalisarpankho

Oriya : Sroetokolothiya

2.3.2. Distribution

Throughout the plains of India, Ceylon, Mauritius, tropical Africa.

2.3.3. Description

Subshrub to 1m; branchlets sericeous – villous. Leaves to 6 cm;

leaflet 5 – 7 pairs, obovate – oblanceolate, 1.5 – 2 × 0.4 – 0.8 cm,

chartaceous, glabrous above, sericeous below, base cuneate, margin entire,

apex obtuse, retuse; petiole to 1cm; petiolule 2 mm; stipules subulate, 3

mm. Pseudoracemes axillary, to 7 cm, flowers paired on rachis; pedicel to

4 mm. Flowers 1 cm across. Calyx – tube 1 mm, sericeous; lobes

17

lanceolate, setaceous, ciliate; upper lobes 4 mm; lower one 5 mm. Corolla

pink; standard orbicular, 1 × 1 cm, sericeous; wings oblong – obovate, 1 ×

0.4 cm; keels 8 × 4 mm, beaked. Staminal sheath 6 mm, sericeous,

continuous within, apex slightly curved, not horned; seeds ca. 8,

subquadrate, 3.5 mm, strophiolate (Mathew, 1983).

2.3.4. Useful Parts

Leaves and root.

2.3.5. Medicinal Uses

Juice of leaves given in dropsy and fresh roots considered hypoglycaemic.

(Anonymous, 1986).

2.4. DISEASES AND PESTS

Amerjothy (2000) highlights the correlations of the loci of the gall-

larva in the cortex, secondary xylem, pith and eventual behavioral patterns

of the vascular cambium. The gradient of cecidogenetic impulses emitted

by the feeding activity of the larva seems to be the key factors for the

differential behavior of the cambium in the shoot-axis galls. Insects

associated with and their larva positions nave been identified.

2.5. ETHNOMEDICINE

Juice of the I. tinctoria leaves used for hydrophobia. Leaf juice and

paste apply on the wound, dogs bite and snake bite. 10ml leaf extract

mixed with honey and drink for curing jaundice and spleen swelling. Fruit

18

used to cure constipation. Root is used in inflammation of the liver,

nervine tonic and antidote for poisoning. (Maheshwari, 1996)

The tribals use the readily available local plants to anual the snake poison.

The species including I. tinctoria often used. (Masilamani, 1997)

The whole plant of I. tinctoria is crushed into a paste, mixed with water

and given to the dog and to detect rabies. If the dog has rabies, infection

will express the symptoms soon. (Mini and Sivadasan, 2007)

Khanna et al. (1994) collected information on unreported medicinal uses

of eighteen plant species belonging to sixteen families exploited as

aphrodisiac among the fork- lores of Uttar Pradesh plains.

Ethnomedical investigation of traditional uses of medicinal plants

by the tribal inhabiting the forest areas of Uttar Pradesh have brought to

light some plants which are utilized for treatment of various skin diseases,

boils and blisters (Singh and Prakash, 1996).

Plant of T. purpurea is tonic, laxative, anthelmitic to children

given to purify the blood and as cordial, decoction is tonic. Root is bitter

chewed to cure colic pain, used in asthma. Juice is mixed with molasses

and given for stomach pain applied on skin eruptions. Powder is smoked

for cough, asthma and respiratory diseases, as paste applied on belly to

19

cure dyspepsia, powdered and boiled in milk is applied on leprosy and

wounds. (Maheswari, 2000)

Hiremath et al. (2007) collected sixty indigenous medicinal plants

from Dharwad and its surroundings. The traditional uses of these plants

were given after discussion with local healers and experienced adults. For

each medicinal plant including T. purpurea, the scientific name, its

family, local name, medicinal use and method of preparation or

administration have been given. The same work had been done on 47

different medicinal plants by Sudha Rani et al. (2007)

Based on the ethno botanical studies T. purpurea roots used as an

ailment for fractures, gas troubles and gastritis. (Lancelot, 2007)

2.6. PHARMACOGNOSY

During literature survey it was found that nine botanically different

species are also called as “Shankhapushpi” in vernacular. Out of these,

four species viz, Canscora diffusa, Clitoria ternatea, Evolvulus

alsinoids and T. purpurea are sold as “Shankhaspushpi” in the area of

western Maharashtra. Botanical details of all these plant species and their

recorded medicinal virtues are given along with; an artificial key is given

to separate all the above mentioned species exomorphically. (Upadhyay

and Kumbhojkar, 1993)

20

Random amplified polymorphic DNA (RAPD) marker was used to

establish intergeneric classification and phylogeny of the tribe Millettieae

sensu Geesink (1984) (Leguminosae:Papilionoideae) and to assess genetic

relationship between 9 constituent species belonging to 5 traditionally

recognized genera under the tribe. DNA from pooled leaf samples was

isolated and RAPD analysis performed using 25 decamer primers. The

genetic similarities were derived from the dendrogram constructed by the

pooled RAPD data using a similarity index, which supported clear

grouping of species under their respective genera, inter- and intra-generic

classification and phylogeny and also merger of Pongamia with Millettia.

Elevation of T. purpurea var. pumila to the rank of a species (T. pumila)

based on morphological characteristics is also supported through this

study of molecular markers. (Acharyaa et al., 2004)

The genotypic variability among twelve species of the genus Tephrosia,

distributed in Andhra Pradesh, through DNA fingerprinting using RAPD

technique. Twenty OPC primers were used. The cluster analysis based

on the similarity matrix was performed using the PHYLIP software

ver.3.65 pooled from all the six primers. It has justified to a great extent

co-relating with the classification based on the morphological traits. It

represents the first approach in using nuclear DNA finger print markers as

a tool to study molecular systematic of the genus Tephrosia. (Lakshmi et

al., 2008)

21

2.7. PHARMACOLOGY

2.7.1. Leaves

Naraikulam et al. (2010) aimed at assessing the protective effect of

I. tinctoria on Carbon Tetrachloride (CCl4) induced hepatotoxicity. The

leaf extract of I. tinctoria increased the antioxidant potential and

hepatoprotective effect of the animal. In histopathological profile, CCl4

treated rat shows hepatocellular necrosis, fatty degeneration and extensive

vacuolation. The treatment with I. tinctoria extract enhances the recovery

from CCl4 induced hepatic damage due to its antioxidant and

hepatoprotective property.

Jain et al. (2006) showed that the ethanol extracts of leaves and

flavonoid (isolated from leaf extract) from T. purpurea were evaluated for

hepatoprotective activity in rats by inducing hepatotoxicity with carbon

tetrachloride. Serum level of transaminases, alkaline phosphate and total

bilirubin and histopathological changes in liver were used as biochemical

markers of hepatotoxicity. Hence, these indicated that the

hepatoprotective activity was more in ethanolic extract of leaves than that

in isolated flavonoid.

The maximum free radical scavenging activity of ethanolic extract T.

purpurea was made the basis of selection of in vivo study. The extract

showed significant reduction in the elevated blood urea and serum

creatinine. Histopathological changes and in vivo antioxidant activity were

determined in line with biochemical findings. The ethanol extract of

22

T. purpurea leaves possesses marked nephroprotective and curative

activities without any toxicity. The proposed mechanisms of activities are

antioxidant activity and inhibition of overproduction of NO and Cox-2

expression and it may be attributed to phenolic and flavonoidal

compounds like quercetin. (Jain and Singhai, 2009)

2.7.2. Seed

Pavana et al. (2007) and Pamu Pavana et al. (2008) investigated that the

beneficial role of T. purpurea ethanolic seed extract on glycoprotein

components in streptozotocin induced diabetic and antilipidperoxidative

rats. Blood glucose and plasma insulin were measured and glycoprotein

components (protein bound hexose, protein bound hexosamine, fucose

and sialic acid) in plasma, erythrocyte membrane, liver and kidney were

investigated and hyperglycemia associated with altered hexokinase and

glucose 6 phosphatase activities, elevated lipid peroxidation, disturbed

enzymatic and non-enzymatic antioxidants status were observed in

streptozotocin induced diabetic rats. Oral administration indicates that

TpEt has potent role in modifying altered glycoprotein components in

streptozotocin induced diabetic rats.

Pavana et al. (2009) evaluated the effects of aqueous seed extract of T.

purpurea (TpASet) on blood glucose and antioxidant status in

streptozotocin induced diabetic rats. Hyperglycemia associated with an

altered hexokinase and glucose-6-phosphatase activities, elevated lipid

peroxidation, disturbed enzymatic and non enzymatic antioxidant status

23

were observed. “TpASet” has potent antihyperglycemic and antioxidant

effects and therefore further studies are warranted to isolate and

characterize the bioactive principles from “TpASet”.

2.7.3. Stem

A new butenylflavanone, a new rotenoid, three active flavonoids and nine

known inactive compounds were isolated from an ethyl acetate-soluble

extract of the stems of T. toxicaria, using a bioassay based on the

induction of quinone reductase (QR) in cultured Hepa IcIc 7 mouse

heptoma cells to moniter chromatographic fractionation. Selected

compounds were tested in a mouse mammary organ culture assay to

evaluate the inhibition of 7, 12-di-methylben {a} anthracene (DMBA)-

induced preneoplastic lesions. (Jang et al., 2003)

2.7.4. Root

Deshpande et al. (2002) reported that the aqueous extracts of roots

of T. purpurea was studied using various ulcer models in albino rats.

Results suggest that AETP possesses significant antiulcer property which

could be either due to cytprotective action of the drug or inhibition of acid

secretion.

Kavitha and Manoharan (2006) investigated that the chemopreventive

potential and antilipidperoxidative effects of ethanolic root extract of T.

purpurea (TpEt) on 7, 12-dimethylbenz (a)anthracene (DMBA) induced

hamster buccal pouch carcinoma. TpEt has potent chemopreventive

24

efficacy and significant antilipidperoxidative effect, in DMBA induced

oral carcinogenesis. Further studies are needed to isolate and characterize

the bioactive principle.

Anti-hyperglycemic activity of roots of T. villosa was evaluated by

Balakrishnan et al. (2007). Aqueous extract of T. villosa were

significantly superior in reducing blood sugar on long treatment.

Anti-implantation activity was lead out by Rajan et al. (2007) on

methanolic extract of Schleichera oleosa (stem bark) and T. purpurea

(root) were screened and both the doses reduced the number of

implantation sites significantly and a dose-related response was observed.

Roots of crude drug of T. purpurea extracted successively using various

solvents [Pet-Ether (60-800C), 95% ethanol and aqueous alcohol (60%

water + 40% ethanol). Single dose administration of all the extracts of T.

purpurea did not showed any hypoglycemic or anti-diabetic activity.

Repeated dose administration of alcoholic and hydro-alcoholic extract

showed significant hypoglycemic and antidiabetic activity at the end of 7th

day. These results suggest that aqueous and hydro-alcoholic extract

possess antidiabetic activity. (Joshi et al., 2008)

Aqueous extract of the roots of T. purpurea was evaluated by Swathi et

al. (2008) for its antilithiatic activity, in two models of urolithiasis.

Gentamicin (s.c.) and 5% ammonium oxalate mixed with rat feed was

25

used to induce calcium oxalate stones; the foreign body implantation

model which makes use of zinc discs, was used to induce magnesium

ammonium phosphate stones. The effect of aqueous extract of T.

purpurea on the excretion and deposition of various calculi forming

constituents like calcium, oxalate, magnesium and phosphate in urine,

kidney and foreign body. The aqueous extract of T. purpurea was found

to be effective in reducing the formation of and dissolving existing

calcium oxalate and magnesium ammonium phosphate stones.

2.7.5. Aerial parts

The alcoholic extract of the aerial parts of I. tinctoria depicted marked

protection against carbon tetrachloride induced liver damage and

histopathological activities in rabbits, rats and mice. The extract also

increased the bile flow and liver damage in rabbits, rats and mice. This

effect was more pronounced in male rats than in female rats. (Anand

et al., 1979 and 1981)

The aerial parts of I. tinctoria extracts showed hypoglycaemic effects in

rats and CNS depressant effects and potentiation of pentobarbitone

induced hypnosis in mice. The LD50 of the extract was �1000 mg/Kg i.p.

in mice. (Dhawan et al., 1980 and Anand et al., 1981)

Singh et al. (2001) experimented a bioactive fraction, indigtone (FA)

obtained by fractionation of a petroleum ether extracts of the aerial parts

of I. tinctoria, showed significant dose related hepatoprotective activity

26

against CCl4 induced liver injury in rats and mice. Hexobarbitone induced

‘sleeptime’, zoxazolamine induced ‘paralysis time’ levels of

transaminases, bilirubin and total protein in serum were employed as

indices of liver injury. Pre post treatment with FA indicated the preventive

restorative effect of FA in the process of CCl4 induced liver damage. The

fraction possessed a high therapeutic ratio, as no mortality was observed

up to a dose of 2 g/Kg p.o. in mice.

T. purpurea was evaluated for its efficacy in rats by inducing

hepatotoxicity with D-galactosamine HCl (acute) and carbon tetrachloride

(chronic). T. purpurea (aerial parts) powder was administered orally and

serum levels of transaminases (SGOT and SGPT) bilirubin and

histopathological changes in the liver were used as the biochemical

markers of hepatotoxicity. The administration of T. purpurea along with

the hepatotoxins offered a protective action in both acute

(D-galactosamine) and chronic (CCl4) models. (Murthy and Srinivasan,

1993)

Gokhale and Saraf (2000) showed the ethanolic extract of dried aerial

parts of T. purpurea inhibited passive paw anaphylaxis in rats and also

passive peritoneal anaphylax is showing antiallergic activity. Significantly

reduced an elevated WBC count in response to antigen challenge in

sensitized mice. The extract also significantly inhibited eosinophil

infiltration without any significant change in the mononuclear cell

population. The extract failed to alter neutrophil adhesion to nylon fibres.

27

It produced a significant inhibitory activity on enzyme lipoxygenase. The

inhibitory effect of ethanolic extract of T. purpurea on late-phase allergy

could be attributed to the inhibition of leukotriene synthesis.

Wound healing potential of T. purpurea L. was experimented by

Lodhi et al. (2006) by using ethanolic extract of aerial parts in the form of

simple ointment using three types of wound models in rats as incision

wound, excision wound and dead space wound. Histopathological study

showed significant (Pless than 0.05) increase in fibroblast cells, collagen

fibres and blood vessels formation.

2.7.6. Entire plant

Progress in the study of traditional Chinese drugs in China has been

presented by Zhang, 1996. Active components obtained from plants and

their pharmacology has been discussed e.g. Antitumour drugs have been

obtained from Taxus bravifloia and I. tinctoria.

The effect of pre-treatment with alcoholic extract of whole plant of

I. tinctoria on liver antioxidant defense endotoxin was investigated in

male albino rats. I. tinctoria pre-treated rats showed considerable

protection against D.Gal N/endotoxin induced oxidative stress as

evidenced by a significant increase in the activities of all the antioxidant

enzymes studied and significant decrease in the level of lipid peroxides.

(Malarvannan and Devaki, 2003)

28

The methanol extract of the whole plant of I. tinctoria was

evaluated by Oli et al. (2005) for its anti-inflammation. It exhibited

significant activity.

Methanolic extract of I. tinctoria exhibited antitumour activity in

Dalton’s Ascitic Lymphoma (DAL) bearing mice. The extract decreased

the lifespan of DAL bearing mice in a day dependent manner. Red blood

cell count, hemoglobin content and white blood cell count were more or

less normal after extract treatment of the tumour bearing mice. (Kavimani

et al., 2001)

The release of LDH and levels of urea in the liver effluent

perfusage, was studied and the rate of bile flow was monitored. Perfusion

with D-galactosamine or CCl4 resulted in increased LDH leakage,

decreased urea levels in the liver effluent and reduction in bile flow.

I. tinctoria pretreatment in vitro ameliorated D-gal N and CCl4 induced

adverse changes towards near normal and thereby indicates its

hepatoprotective effects in rats. (Sreepriya et al., 2001)

Ten medicinal plants including alcohol dried extract of T. uniflora

were pharmacologically screened for their cardiac activity on isolated

rabbit heart and showed significant negative inotropic activity with

negative chronotropic effects. In all cases alcohol dried extracts were used

29

and reconstituted in either water or ethyl acetate for tests. The plant

extracts which produced cardiac depression (Sajeed et al., 1996).

Dutta et al. (1997) worked on “Yakrifit" (a polyherbal product in

which T. purpurea is a part) that was administered to all the animals

either as liquid 25-30 ml orally twice daily for 3-6 days or as one bolus

twice daily for 5 days. All the animals recovered in 3 to 7 days, regained

appetite for food and water and their general condition had improved.

The therapeutic efficacy of an herbal hepatoprotective

hepatostimulant AV/LTP/15 (a polyherbal product consisting of

T. purpurea) oral in removing anorexia syndrome in dogs is reported by

Singh (1997) and Agarwal (1999). AV/LTP/15 gave well to very good

response in 82.1 percent of the dogs. The formulation was quite palatable

and well-tolerated by dogs of all ages.

The protective effect of HD-03 was observed in all three types of

intoxication, which are different in their primary mechanism of inducing

hepatotoxicity, a protective mode of action of HD-03, not specific to the

hepatotoxic has been suggested by Mitra et al. (1998).

Anticholestatic acitivity was investigated by Mitra et al. (1999) that HD-

03, a herbal formulation containing T. purpurea as one of the ingredients

in thioacetamide (TAA)- induced experimental cholestasis in

anaesthetized guinea pigs which significantly prevented thioacetamide

30

induced changes in bile flow, bile acids and bile salts excretion. HD-03

has been suggested to serve as a potent choleretic and anticholestatic

agent.

Therapeutic uses of T. purpurea as mentioned in Ayurvedic texts,

pharmacological activity and chemical constituents isolated from the plant

have been reviewed by Patel et al. (1999).

Lucas and Ananada Rajasekhar (2000) reviewed experimental

studies on anti-hepatotoxic activity of certain medicinal plants including

T. purpurea used in Ayurveda. Out of those medicinal plants,

Boerhaavia diffusa exhibited cell regeneration effect and has a better

anti-hepatotoxic effect in comparison with other medicinal plants.

Saleem et al. (2000) assessed the topical application of

T. purpurea 1h prior to each application of croton oil (phorbol ester)

resulted in a significant protection against cutaneous carcinogenesis in a

dose-dependent manner. The animals pre-treated with T. purpurea

showed a decrease in both tumor incidence and tumor yield. A significant

reduction in TPA (12-O-tetradecanoyl phorbal-13-acetate) mediated

induction in cutaneous ornithine decarboxylase (ODC) activity and [H3]

thymidine incorporation was also observed. The topical application of

T. purpurea prior to TPA resulted in the significant recovery of

TPA-mediated depletion in the level of glutathione, glutathione

S-transferase, glutathione reductase and catalase. T. purpurea can

31

abrogate the tumor-promoting effect of croton oil (phorbol ester) in

murine skin.

The protective role of T. purpurea has been investigated by Kumar et al.

(2001) the alcohol extract of T. purpurea showed a significant hydroxyl

radical scavenging activity in vitro. Using a Trypan blue exclusion assay,

it was found that the extract markedly increased the percentage viability of

the isolated rat kidney cortical cells in gentamicin-induced cell damage.

By the evaluation of LDH activity and acid phosphatase content, it was

established that the cell damage was minimized in the case of cells treated

with the extract of T. purpurea. The hydroxyl radical scavenging effect of

the extract was enhanced with increases in the concentration of drug,

suggesting the role of free radical scavengers in minimizing kidney cell

damage.

Taraphdar et al. (2002) examined the activities of T. purpurea in radiation

induced haemopoietic injury to total body irradiation. It induced

significant increase in haemoglobin and total RBC count. After

irradiation there was no fall in RBC count and haemoglobin and

Tephrosia has a selectively effective on erythroid compartment.

A fraction (F062) obtained from n-butanol extract of T. purpurea

showed consistent antileishmanial activity against Leishmania donovani

32

infection in hamsters. Activity was further confirmed in a secondary

model, i.e., Indian languor monkeys (Presbytis entellus). The fraction

F062 from this plant possesses potential to produce significant

antileishmanial activity by oral route without producing any toxic side

effects. (Sharma et al., 2003)

Soni et al. (2003) stated that, the dried alcoholic extract of

T. purpurea was investigated for its free radical scavenging activity,

antilipid peroxidation potential and hydroxyl radical scavenging activity

and significant inhibition of lipid peroxidation. However, it failed to show

any significant scavenging effect on hydroxyl radicles.

The flavonoid fraction of T. purpurea (FFTP) was studied for its

effect on cellular and humoral functions and on macrophage phagocytosis

in mice. Oral administration of FFTP (10–40 mg/Kg) significantly

inhibited sheep red blood cells (SRBC)-induced delayed-type

hypersensitivity reactions. It also produced a significant, dose-related

decrease in sheep erythrocyte-specific haemagglutination antibody titre.

(Damre, 2003)

Cao et al. (2004) reported that the bioassay guided fractionation of

methanolic extracts of Mundulea chapelieri resulted in the isolation of

two new flavonoids and eight known flavonoids which containing

rotenolone, retoenone and tephrosin. All the isolated compounds were

tested cytotoxicity against the A 2780 human ovarian cancer cell line,

33

rotenolone and rotenone were the most potent compounds isolated, with

IC50 values of 0.5 and 0.7 micr/mL, respectively.

Anti-inflammatory activity of a flavonol glycoside from T. spinosa

was investigated by Chakradhar et al. (2005) and its activity was

evaluated against carrageenin induced paw edema. The compound

exhibited significant activity.

Prabhu Nair (2006) showed protective effect of Tefroli tonic (a

polyherbal mixture containing T. purpurea) against cadmium induced

hepatotoxicity in experimental rats. Subcutaneous injection of cadmium

chloride to rats caused liver damage and was observed by analysis of

serum bilirubin and assay of marker enzymes such as trnsaminase and

phosphates of body serum and liver. The administration of Tefroli tonic

has maximum protective effect against cadmium chloride induced

hepatotoxicity in rats.

The ethanol extract of T. purpurea Linn. was found to significantly

inhibit the carbon tetrachloride-induced lipid peroxidation in vivo and

superoxide generation in vivo. The ethyl acetate fraction of the same

extract was studied for free radical scavenging and antilipid peroxidation

activity. The IC50 values in both of these in vitro assays were found to be

34

significantly reduced for ethyl acetate fraction compared with the

ethanolic extract of the plant. The observation was further supported by

comparing the in vivo antioxidant activity for both the ethanolic extract

and its ethyl acetate fraction. The ethanolic extract of T. purpurea exhibits

antioxidant activity in vivo and the ethyl acetate soluble fraction has

improved antioxidant potential than the extract. (Soni et al., 2006)

Kishore kumar et al. (2007) concluded that significant hepatoprotective

effect was obtained against CCl4 induced liver damage, by oral

administration of methanolic extract of T. falciformis as evident from

decreased levels of serum enzymes in the treated groups, compared to the

controls.

Oral administration of T. purpurea and Tecomella undulata

resulted in a significant reduction in serum aspartate aminotransaminase,

alanine aminotransaminase, gamma glutamyl transpeptidase, alkaline

phosphatas, total bilirubin and liver MDA levels and significant

improvement in liver glutathione when compared with thioacetamide

damaged rats. Histology of the liver sections of the animals treated with

the extracts also showed dose-dependent reduction of necrosis. (Khatri

et al., 2008)

35

Khan et al. (2008) investigate a chemopreventive efficacy of

T. purpurea against N-diethylnitrosamine-initiated and potassium

bromate-mediated oxidative stress and toxicity in rat kidney. The

susceptibility of renal microsomal membrane for iron ascorbate-induced

lipid peroxidation and xanthine oxidase activities were significantly

reduced. The depleted levels of glutathione, the inhibited activities of

antioxidant enzymes, phase II metabolizing enzymes and the enhanced

levels of serum creatinine and blood urea nitrogen were recovered to a

significant level. All the antioxidant enzymes were recovered dose-

dependently. T. purpurea besides a skin antioxidant can be a potent

chemopreventive agent.

Polyherbal formulation available with a wide range of indications like

protective to liver, appetite and growth promoters, gastrointestinal and

hepatic regulator, as treatment for hepatic dysfunction, for hepatic

regenerations as well as liver stimulant and tonic. To evaluate the

hepatoprotective activity of six commercially available formulations,

namely Liv 52, Livergen, Livokin, Octogen, Stimuliv and Tefroli in acute

toxicity in mice model induced by Paracetamol (PCM). (Girish et al.,

2009)

2.8. PHYSIOLOGY AND BIOCHEMISTRY

Plant tissue concentrations of various elements were typical of species

growing on ultramatic substrates. A usual finding was the discovery of T.

villosa is one of the five hyper-accumulators of copper (more than 1000

36

micro g/g dry tissue) Copper hyper accumulation in ultramatic substrate is

a little known phenomenon and requires further study (Rajakaruna and

Bohm, 2002).

Bhaskar Rao et al. (2007) evaluated in vitro antioxidant properties of

aqueous extracts of Alternanthera sessilis and T. purpurea. Both the

plants are beneficial as an antioxidant sources and the plants possess

significant levels of enzymatic antioxidants, non-enzymatic antioxidants

and also exhibits antioxidant capacity.

2.8.1. Leaves

Lambert et al. (1993) investigated that the rotenoid compounds provided

from the phenylproponoid pathway are largely accumulated in the leaves

of T. vogelii. The photomixotrophic cell line accumulated rotenone and

deguelin in Tephrosia leaves and the heterotrophic cell line produced

essentially deguelin and tephrosin.

The in vitro antioxidant activity of ethanolic extract of leaves of

T. purpurea was investigated by DPPH free radical scavenging and nitric

oxide scavenging methods. The ethanol extract showed good antioxidant

activity in these above methods. This activity may be due to the presence

of flavonoids (Jain et al., 2007).

2.8.2. Seed

Nagarajan and Merita (2001) stated that the twenty five seeds of

T. falciformis were given various treatments to establish the germination

37

requirements. Seed treatment significantly influenced germination ranged

from 1.7-25.0 and lowest value was recorded in seeds soaked in cold

water treated for 12 hours and the highest in mechanically scarified seeds

followed by seeds soaked in cold water for 6 hours.

Zafar et al. (2002) studied germination of T. purpurea seeds. The

mechanical treatment i.e., removal of seed coat at one end of the seed and

its subsequent treatment with Ethrel (1000ppm) gave 100 percent

germination within 5 days while the seeds without this treatment took

12-20 days to germinate and the rate of germination observed was 10

percent.

2.9. TISSUE CULTURE

The total rotenoid content of I. tinctoria decreased with age;

among the plant parts, maximum content was in leaves and minimum in

stem. The identity of different rotenoids was confirmed by melting point

mixed melting (deguelin, dehydrodeguelin, rotenol, rotenone, tephrosin

and sumatrol) were isolated identified and quantified in vivo. Only four

rotenoids were present in callus cultures; sumatrol and tephrosin were

absent. The toxicological studies of in vivo and in vitro extract against the

pulse beetle (Callosobruchus chinensis) and mosquito (Anopheles

stephensi) larvae showed that rotenoids were more effective against

mosquito larvae than C. chinensis. Extracts from callus was more

38

effective against both the test animals than that from plant parts. (Kamal

and Mangla, 1993)

Kumar and Saxena (2003) isolated secondary metabolites (flavonoids)

from 13 species including I. tinctoria.

Lambert et al. (1993) provided rotenoid compounds from the

phenylpropanoid pathway are largely accumulated in leaves of T. vogelii.

Heterotrophic and photomixotrophic cell suspension cultures of this

tropical plant have been obtained. Both cell lines are able to produce

rotenoids, but a specific production is observed in each cell culture. The

photomixotrophic cell line accumulated rotenone and deguelin in

Tephrosia leaves and the heterotrophic cell line produced essentially

deguelin and tephrosin.

Zafar and Mujeeb (2002) cultured root, stem and leaf of

T. purpurea in tissue culture and successfully developed and maintained

on Murashige and Skoog’s medium supplemented with various plant

growth regulators. The content of rotenoids and rutin in the callus cultures

were estimated by spectrophotometric method.

2.10. PHYTOCHEMISTRY

2.10.1. Leaves

39

From dried leaves of T. spinosa, eupalitin 3-O-beta-D-galactopyranoside

(yield 0.05 percent) was isolated and identified by Vanangamudi et al.

(1997).

From the methanolic extracts of leaves of T. purpurea (+)– Tephrosin has

been derived by Nurdjaman et al. (2007). (±)–Tephrosin has been shown

as insecticide against silkworm through bioassay and also shown activity

as inhibitor against phrobol mouse 308 epidermal cells and as cancer

chemopreventive agent last tephrosin shown as fish poison against gold

fish (Cyprinus carpio).

40

2.10.2. Stem

A novel neoflavonoid glycoside, serration 7-O-{beta-D-glucopyranosyl-

(1 to 4) –beta-D-galactopyranoside} (C27H30O142 mp 168-70 degree C)

was isolated from the stem of T. purpurea and identified by its chemical

and spectral analysis. (Saxena and Choubey, 1997)

Jang et al. (2003) isolated a new butenylflavanone, (2S)-5-hydroxy-7-

methoxy-8-[(E)-3-oxo-1-butenyl] flavanone (1), and a new rotenoid,

4‘,5‘-dihydro-11,5‘-dihydroxy-4‘-methoxytephrosin (2), as well as three

active flavonoids of previously known structure, isoliquiritigenin (3),

genistein (4), and chrysoeriol (5), along with nine known inactive

compounds, α-toxicarol (6), sumatrol, 6a,12a-dehydro-α-toxicarol, 11-

hydroxytephrosin, obovatin, marmesin, lupenone, benzyl benzoate, and

benzyl trans-cinnamate, were isolated from an ethyl acetate-soluble

extract of the stems of T. toxicaria, using a bioassay based on the

induction of quinone reductase (QR) in cultured Hepa 1c1c7 mouse

hepatoma cells to monitor chromatographic fractionation. The structures

of compounds 1 and 2 were elucidated by spectroscopic data

interpretation. All isolates were evaluated for their potential cancer

chemopreventive properties utilizing an in vitro assay to determine

quinone reductase induction. Selected compounds were tested in a mouse

mammary organ culture assay to evaluate the inhibition of 7, 12-

dimethylbenz[a]anthracene (DMBA)-induced preneoplastic lesions.

41

2.10.3. Stem and Leaves

Elongatin (mp 182-83 degree C), 120-hydroxyrotenone, beta-sitosterol

and stigmasterol have been isolated and identified from the stems and

leaves of T. uniflora. (Abreu and Luis, 1996).

Aruna et al. (1999) established that the stem and leaves of

T. procumbens yielded seven flavonoid compounds – obovatin, sumatrol,

rotenone, maxima isoflavone A, maxima isoflavone J, maxima isoflavone

C and fisetin 7-ethyl ether by chromatographic techniques.

Arriage et al. (2005) obtained essential oils from fresh leaves and

stems of T. egregia and were analyzed by GC and GC/MS. Nine and 10

volatile compounds have been identified in the leaf and stem oils

respectively. The major components of the leaf oil were geijerene (50.3

percent) and pregeijerene whereas the stem oil contained geijerene and

pregeijerene as main constituents.

2.10.4. Seed

A total of six complex 7-oxygenated-8-prenylflavones have been

isolated from the seeds of T. apollinea and identified as the

diastereoisomers (−)-semiglabrin and (−)-pseudosemiglabrin, (+)-

glabratephrin, (+)-glabratephrinol, appollinine (7-methoxy-8-[3″-(2″, 5″-

dihydro-5″, 5″-dimethyl-2″-oxofuryl)]-flavone and lanceolatin-A. The use

of 13

C NMR in the structure elucidation of flavones of this type is

discussed by Waterman and Khali, 1979.

42

Two prenyated flavonoids purpuritenin and purpureamethide have

been characterized by Bharathi Sinha et al. (1982) from the seeds of

T. purpurea.

Parmar et al. (1988) reported that the isolation from the seeds of

T. candida and the separation of tephrosin and 12a-hydroxyrotenone by

hplc; caffeic acid, β-sitosterol, and its glucoside were also isolated. These

compounds have been isolated for the first time from this genus, and the

mixture of tephrosin and 12a-hydroxyrotenone has been resolved for the

first time using hplc.

Horie et al. (1994) revised the structure of a natural flavone from seeds of

T. candida which assumed to be 5,4’- dihydroxy-3,6,8-trimethoxy

flavone, and renamed as 5,4’-dihydroxy-3,6,7-trimethoxyflavone,

penduletin on the basis of spectral data.

Isopongaflavone isolated from the seed of T. bracteolata has been

synthesized from phloroacetophenone in six steps by Amzad Gissaub and

Tarafdar (1998).

Fatty acid compositions of seed oils of eight species including

T. purpurea are reported by Deora et al. (2003). All the seeds were

collected from arid zone of Rajasthan state and examined for their

physico-chemical characteristics and fatty acid composition.

43

2.10.5. Root

Lwande et al. (1988) reported that the isolation and identification of six

flavonoids from the roots of T. elata. They have been identified as the

flavanones 8-(3, 3-dimethylallyl)-5, 7-dimethoxyflavanone and obovatin

methyl ether, the flavones warangalone (scandenone), the pterocarpans

(+)-pisatin and (-)maackiain, and the rotenoid tephrosin. Although

isopongaflavone was found to occur in large quantities (1.2%) in the seeds

of T. elata.

A new flavanone, maxima flavanone A, was isolated by Rao et al.

(1994) from the chloroform extract of the roots of T. maxima along with

maxima isoflavone J and their structures established from spectral data.

Rao and Ranga Raju (1995) isolated steroid constituents of

chloroform extracts of T. purpurea root and identified as: stigmastrol,

beta-sitosterol, campesterol, stigmasta-4-en-3-one and stigmasta-4-, 22-

dien-3-one.

The hexane and ethyl acetate extracts of roots of T. candida

afforded a new rotenoid, 12a-hydroxy-beta-toxicard, together with a series

of known ones, identified as deguelin, alpha-toxicarol, tephrosin, 6a, 12a-

dehydrodeguelin, 12a-hydroxy-alpha-toxi carol, 6a, 12a-dehydro-alpha-

toxicard and 6a, 12a-dehydro-beta-toxicarol. The possibility of four of

these rotenoids to be linear or angular is discussed on the basis of HMBC

experiment (Andrei et al., 1997)

44

Tarus et al. (2002) stated that from the petrol extract of the roots of

T. aequilata, five flavonoids were isolated of which, 3, 4: 8, 9 –

dimethylenedioxy pterocarpan is reported for the first time.

Ganapathy et al. (2003) isolated rotenone, obovatin, sumatrol,

praecansone B, praecansone A, fisetin 7-ethyl ether and two new

isoflavones. Calopogonium isoflavone B (C21H6O5; mp 213 degree C) and

2’,7,8-trimethoxy-4’,5’-methylenedioxy isoflavone yield 0.01 percent,

(C19H16O7, mp 204-06 degree C) from the roots of T. procumbens.

Chemical examination of the roots of T. tinctoria led to the

isolation of β–sitosterol, lupeol, tephrinone, 7-O-methyl glabranin,

2-Hydroxy tephrosin, rotenone and dehydrodegulin. The compounds were

isolated by sequential chromatography and the structures were established

by 2D NMR analysis and MS spectral data. All the compounds are

reported for the first time from the species T. tinctoria and the candidate

2-Hydroxy tephrosin is new to the species and also to the genus Tephrosia

as well. (Lakshmi et al., 2008)

2.10.6. Aerial parts

Vanangamudi et al. (1997) derived that the chloroform extract of

the aerial parts of T. hookeriana resulted in the isolation of a new flavone

named tephrorianin (mp.228-30 degree C) and characterized as 2”,3”-

dihydro-5-methoxy-3”-(2”-acetoxy-2”-methylpropylidene)-2”-oxofuryl-

45

(4”,5”; 7,8)- flavone along with (-)-semiglabrin and lanceolatin A from

the pods, hookerianin from the stem and rutin from the leaves.

NMR spectra of tephrosin (mp.198-200 degree C) pongaglabol (mp.196-8

degree C) and semiglabrin (mp.253-4 degree C) isolated by Ahmed et al.

(1999) from T. purpurea aerial parts are reported.

Mohamed-Elamir et al. (2009) reported that the chemical investigations of

aerial parts of T. purpurea yielded the rare prenylated flavonoids,

tephropurpulin A and isoglabratephrin in addition to previously identified

flavonoids glabratephrin. Structures were established by 1D and 2D NMR

spectroscopy, as well as by HR-MS analysis; for compounds 2 and 3,

structures were confirmed by X-ray analysis.

2.10.7. Roots and Aerial parts

Gomez-Garibay et al. (1997) reported that the roots and aerial parts

of T. tepicana afforded a prenyl biflavanol. The structure and

stereochemistry were established by spectroscopic methods some

chemical transformations and confirmed by X-ray diffraction. Tepicanol

A is the first biflavanol with a 4, 4” biflavanyl ether group to be isolated

from the genus Tephrosia.

46

2.10.8. Entire plant

Progress in the study of anticancer drugs originating from plants in China

has been reviewed by Han (1995). Some anticancer drugs have been

found, including indirubin from I. tinctoria.

Modified flavonoids i.e semiglabrin, pseudosemiglabrin were isolated by

Khalid and Narender (2004) from I. tinctoria along with rare flavonoids

glycoside, kaempferol-4-7-dirhmnoside. Their structure elucidation using

modern NMR and other spectral data has been described.

The structure and absolute configuration of lupinifolin, (2S) - 4′,5 -

dihydroxy - 8 - (3 - methyl - 2 - butenyl) - 2″, 2″ - dimethylpyrano[5″.6″ -

g]flavanone, and lupinifolinol, (2R,3R) - 8 - (3 - methyl - 2 - butenyl) -

3,4′,5 - trihydroxy - 2″,2″ - dimethylpyrano[5″.6″ - g]flavanone, have been

deduced from T. lupinifolia. Burch. and spectroscopic, chemical

evidence. (Smalberger et al., 1974)

Pongamol in its pure enol from has been found to occur in the whole plant

for T. purpurea together with β–sitosterol, Ursolic acid and Spinasterol-α

and isolated by Virinder et al., 1989.

An attempt is made to discuss a few traditional medicinal plants used in

skin disease including T. purpurea in respect of their chemical

constituents and medicinal action (Kamil, 1994).

47

Chang et al. (1997) studied that an isoflavone and a chalcone both novel

compounds as well as six constituents of known structure, (+)- purpurin,

pongamol, lanceolatin B, (-)- maackiain, (-) -3-hydroxy-4-methoxy-8-, 9-

methylene-dioxyperocarpan and (-)- medicarpin, were obtained as active

compounds from T. purpurea, using a bioassay based on the induction of

quinone reductase (QR) activity with cultured Hepa /c/c 7 mouse

hepatoma cells. Additionally, three inactive compounds were isolated and

identified.

Sreenivasulu and Sarma (1998) reported that the petroleum ether

extract of the whole plant of T. strigosa contains beta-amyrin,

n-triacontanol, beta-sitosterol, 3R (-) mucronulatol and oleanolic acid.

The relative and absolute stereochemistries of (+) – purpurin, a

flavanone natural product from Tephrosia, were determined to be 2S,7aR,

10S, 10aS by synthesis from semiglabrin in conjunction with x-ray

crystallographic analysis by Pirrung et al. (1998)

Rao and Sridhar (1999) isolated an isoflavan and a pentacyclic

triterpene from the chloroform extract of T. strigosa. This is the first

report of the occurrence of an isoflavan compound in the genus

Tephrosia.

Three novel flavonoids, (+)-tephrorins A (1) and B (2) and (+)-

tephrosone (3), were isolated from T. purpurea. Their structures were

48

elucidated by NMR spectral analysis, and their absolute configurations

were determined by Mosherester methodology. Compounds 1 and 2 are

flavanones containing an unusual tetrahydrofuran moiety. Compounds

1−3 were evaluated for their potential cancer chemopreventive properties

using a cell-based quinone reductase induction assay. (Chang et al., 2000)

From the diethyl ether extract of Sarcolobus globosus (Asclepiadaceae) a

new rotenoid and a new isoflavone as well as previously known rotenoids

tephrosin, 12a alpha-hydroxydeguelin, 11-hydroxytephrosin, 12a –

hydroxyrotenone, 12a alpha-hydroxyrotenone, 6a alpha, 12a alpha-12a-

hydroxyelliptone, 6a,12a-dehydrodeguelin, 13-homo-13-oxa-6a, 12a-

dehydrodeguelin, the isoflavone barbigerone and a chromone 6,7-

dimethoxy-2, 3-dihydrochromene were isolated and identified by

Wangensteen et al. (2005).

Zubairi et al. (2005) standardize and determine the bio-active

compounds from the extract of Derris elliptica using the internal standard

method of the isocratic High Performance Liquid Chromatography

(HPLC) analysis system. The root and stem were extracted using the

Normal Soaking Extraction process at 28oC to 30

oC with 95.0% (v/v) of

Acetone as a solvent-to-solid ratio of the extraction. The employed

method of analysis shows significant appearances of the bio-active

compounds in the extract compared with the commercial grade of

rotenone cube resin.

49

Three rotenoids viz, rotenone, elliptone and deguelin were

identified from various plant parts of Parkinsonia aculeate. L. and using

TLC, GLC, mp, mmp, IR, and UV studies, which were comparable to that

of their respective standard compounds. The different doses (1000 igmL-1

,

500 igmL-1

, 250 igmL-1

, 125 igmL-1

, 62 igmL-1

, and 31igmL-1

) of isolated

rotenoids from roots were subjected to in vitro antiamoebic activity along

with standard rotenone and derris resin for different time intervals. The

dose level 500 igmL-1

was found most active as compared to 250 ig/mL-1

of standards. (Kamal and Mathur, 2007)

Phenolic compounds have been analysed by Subbha Rao and

Shanmukha Rao (2008) in eighteen taxa of Tephrosia. In all, forty one

compounds have been detected of which eight are identified. Further, it

lends support to maintain T. hamiltonii as distinct from T. purpurea and

taxonomic distinction of T. strigosa.

Tannins and Flavonoids present in the Terminalia chebula,

flavonoids like rutin and quercetin possess man biochemical effects like

inhibition of enzymes, regulatory role on different hormones and

pharmacological activities like antimicrobial, antioxidant, and anticancer,

antihepatotoxic, protection of cardio vascular system. An HPLC method

was developed for the estimation of rutin and quercetin from methanolic

extract of T. chebula. (Kumar et al., 2009)

50

51

Dr.Duke's Phytochemical and Ethnobotanical Databases

godwinm001@hawaii.rr.com

Chemicals and their Biological Activities in: T. purpurea PERS.

(Fabaceae)-Purple Tephrosia, Wild Indigo

Chemicals Activities

BETULINIC-ACID

(Plant 30 ppm)

Anthelmintic; Antibacterial; Anticancer;

Anticarcinomic; Antiedemic; AntiHIV

Antiinflammatory;Antileukemic;Antimalarial;

Antimelanomic;Antinociceptive;

Antiplasmodial; Antitumor; Antiviral

Apoptotic; Cytotoxic Phospholipase-A2-

Inhibitor; Prostaglandin-Synthesis-Inhibitor

ISOTEPHROSIN

(Root)

No activity reported.

LUPEOL

(Plant)

Antiangiogenic AntiEBV; Antiedemic; Antiflu;

Antihyperglycemic; Antiinflammatory;

Antilithic Antimalarial Antioxalate Antioxidant;

Antiperoxidant; Antiprostaglandin;

Antirheumatic; Antitumor; Antiurethrotic;

Antiviral; Cytotoxic FPTase-Inhibitor

Hypotensive; Pesticide; TOPO-2-Inhibitor

ROTENOIDS

(Leaf 6,500 - 8,000

ppm)

(Root 7,000 - 9,500

ppm)

(Seed 16,000-

18,000ppm)

(Stem 4,000 - 6,500

ppm)

No activity reported.

ROTENONE

(Root 7,000 ppm)

Acaricide; Antifeedant; Antitumor Bruchicide;

Convulsant; Ectoparasiticide; Insecticide;

Larvicide; Pediculicide; Pesticide; Piscicide;

Pulicide; Scabicide

RUTIN

(Leaf)

5-HT-Inhibitor; Aldehyde-Oxidase-Inhibitor

Aldose-Reductase-Inhibitor Allelochemic;

52

Antiaggregant Antiallergic; Antiapoplectic;

Antiatherogenic; Antiatherosclerotic;

Antibacterial; Anticancer; Anticapillary-

Fragility Anticataract; Anticlastogen;

Anticonvulsant; AntiCVI Antidementia;

Antidermatitic; Antidiabetic; Antiedemic

Antierythemic; Antifeedant; Antiglaucomic

Antihematuric; Antihemorrhoidal;

Antihepatotoxic; Antiherpetic; Antihistaminic;

Antihypertensive; Antiinflammatory

Antimalarial , Antimelanomic; Antimetastatic;

Antimutagenic, Antinephritic; Antinociceptive;

Antioxidant, Antiperoxidant, Antiplatelet;

Antiproliferant, Antiprotozoal; Antipurpuric;

Antiradicular 9 x quercetin; Antispasmodic;

Antisunburn; Antithrombogenic Antithyroid,

Antitrypanosomic, Antitumor; Antitumor-

Promoter; Antiulcer; Antivaricose; Antiviral;

Apoptotic, cAMP-Phosphodiesterase-Inhibitor;

Cancer-Preventive; Capillariprotective;

Catabolic; Chemopreventive; Cytoprotective,

Estrogenic; Hemostat; Hepatomagenic,

Hepatoprotective, Hypocholesterolemic;

Hypotensive; Immunomodulator; Insecticide;

Insectiphile; Juvabional; Larvistat,

Lipoxygenase-Inhibitor, Mutagenic;

Myoprotective; Myorelaxant; Oviposition-

Stimulant; PAF-Inhibitor; Pesticide;

Protisticide; Radioprotective; Sunscreen;

Topoisomerase-II-Inhibitor, Vasodilator;

Vasopressor

SILICA

(Leaf 21,900ppm)

No activity reported.

TEPHROSIN

(Root)

Antifeedant; Cytotoxic; Pesticide; Piscicide

Tephrosin

Tephrosin is a natural fish poison found in the leaves and seeds of

T. purpurea.

53

Other names 12aβ-hydroxydeguelin

Identifiers

CAS number [76-80-2]

PubChem 114909

Properties

Molecular

formula C23H22O7

Molar mass 410.41658 g/mol

Related compounds

Related

compounds Deguelin, toxicarol

Except where noted otherwise, data are given for

materials in their standard state(at 25 °C, 100 kPa) Infobox references

Retrieved from "http://en.wikipedia.org/wiki/Tephrosin

From the roots of T. purpurea, the following compounds are isolated by

Hui ye gen.

Deguelin

Degueline. [522-17-8] C23H22O6 (394.42). Yellow crystals, mp 180-

54

182°C (methanol); 171°C, [α] D20

= -107° (c = 0.2, benzene). Ornithine

decarboxylase inhibitor (induced by ester phorbol, IC50 = 0.0003µg/ml);

larvacide (larva of mosquito); nematocide (MLD = 1µmol/L).

Dehydrodeguelin

6α, 12α-Dhydrodeguelin. [3466-23-7] C23H20O6 (392.41). Straw yellow

solid, mp 215-225°C. cAMP phosphodiesterase inhibitor (rat heart, IC50 =

6.2µmol/L); larvacide (larva of mosquito); nematocide (in vitro, 0.1µg/ml,

larva of Toxocara canis, after 6 hours cultivation,RM=30, after 24 hours,

RM = 0).

(-)-3-Hydroxy-4-methoxy-8-9-methylenedioxypterocarpan

[69626-65-9] C17H14O6 (314.29). Acicular crystals (methanol), mp 156-

159°C, [α] D20

= -166.6° (c = 0.12, chloroform). Induces quinone reductase

(mus hepatic cells Hepa lclc 7µmol/L, CD 14.7µmol/L).

Methylpongamol

O-Methylpongamol. [80158-88-9] C19H16O4 (308.33). Yellowish oil.

Nematocide (0.1mg/ml cultured with Toxocara canis larvas, after 6 hours

RM = 70, after 24 hours RM = 33).

Pongamol

Lanceolatin C. [484-33-3] C18H14O4 (294.41). Yellow prismactic crystals

(methanol), mp 130-131°C. Nematocide (0.1mg/ml, cultured with larva of

Toxocara canis, in 6 hours RM = 0); sedative; LD50 (ip) = 17.14 mg/Kg.

Pseudosmiglabrin

(-)-Pseudosemiglabrin. [75444-25-6] C23H20O6 (392.41). Coloress

lamellar crystals (methanol), mp 171-174°C; mp 181-183°C, [α] D25

= -

55

384° (c = 0.49, chloroform). Platelet aggregation inhibitor (selective,

caused by thromboxane A2, 6.5µg/ml, inhibitive rate = 85±5%, IC50 =

12.5µmol/L).

Rotenolone

Rotenolone I; Rotenolon I; 12a-Hydroxyrotenone; 6aβ,12aβ-Rotenolone;

Rotenalone. [509-96-6] C23H22O7 (410.42). Yellowish solid, mp

88°C. Cytotoxic (KB, ED50 = 0.01-0.30µg/ml); acaricide; nematocide

(0.1mg/ml cultured with larva of Toxocara canis, 3 hours later RM = 33,

6 hours later RM = 0); MLC=5µmol/L.

Rotenone

[83-79-4] C23H22O6 (394.43). mp (-) 163°C. Antiprotozoal; fish toxin;

pesticide; LD (dog iv) = 0.5 mg/Kg; LD50 (mus ip) = 2.8 mg/Kg.

Tephrosin

C23H22O7 (410.43). mp 198°C. Insect antifeedant.

α-Toxicarol

[82-09-7] C23H22O7 (410.43). Yellowish solid, mp 98-102°C; mp (-) 125-

127°C, (±) 219-223°C. Nematocide (in vitro, 0.1mg/ml, larva Toxocara

canis, after 6 h cultivation, RM = 4, after 24 h, RM = 0); pesticide.

2.11. ANTIMICROBIAL ACTIVITY

2.11.1. Seed

Thetwar et al. (2006) revealed that the seed extracts of the plant

T. purpurea were tested for their antimicrobial and antifungal properties

in various solvents against some human, animal and plant pathogenic

56

bacteria. The seed extract showed a good inhibition effect against all the

tested micro-organism.

2.11.2. Root

Deshpande et al. (2005) proved that the fresh extract of

T. purpurea root was treated for antibacterial and antifungal activity by

agar well diffusion and R&B agar well diffusion method respectively.

This extract shows antibacterial activity.

Kumar et al. (2007) conducted to evaluate antimicrobial activities

of Indian medicinal plants against these etiologic agents of Acne vulgaris.

Ethanolic extracts of Hemidesmus indicus (roots), Eclipta alba (fruits),

Coscinium fenestratum (stems), Curcubito pepo (seeds), T. purpurea

(roots), Mentha piperita (leaves), Pongamia pinnata (seeds), Symplocos

racemosa (barks), Euphorbia hirta (roots), Tinospora cordyfolia (roots),

Thespesia populnea (roots), and Jasminum officinale (flowers) were

tested for antimicrobial activities by disc diffusion and broth dilution

methods. The results from the disc diffusion method showed that 07

medicinal plants could inhibit the growth of Propionibacterium acnes.

Among those Hemidesmus indicus, Coscinium fenestratum,

T. purpurea, Euphorbia hirta, Symplocos racemosa, Curcubito pepo and

Eclipta alba had strong inhibitory effects.

57

2.11.3. Aerial parts

Euchrestaflavanone A and abyssinone-V, two flavonoids are isolated from

the dry crude ethanol extracts of the aerial parts of Mundulea monantha

and T. linearis and plants had satisfactory bacteriostatic and bactericidal

activity against Gram-positive strains. (Ratsimamanga-urverg et al., 1994)

Adoum et al. (1998) studied twenty five plants from 20 families were

selected on the basis of a literature survey and uses in Hausa folk

medicine. Samples were extracted, fractioned and screened for bioactivity

against the brine shrimp larvae and 8 microbial pathogens and selective

antimicrobial activity in the extracts of T. prupurea is noteworthy.

The antibacterial activity of five plants was examined by Mahajan

et al. (1999) against E. coli, Proteus vulgaris, Salmonella typhi, Bacillus

subtilis and Staphylococcus aurens. Among the five plants, the ethanolic

and methanolic extracts of T. purpurea and B. roxburghii possessed

potential antibacterial activity.

Vijayan et al. (2008) characterized a defensin, TvD1 from a weedy

leguminous herb, T. villosa. The open reading frame of the cDNA was

228 bp, which codes for a peptide with 75 amino acids. Expression

analyses indicated that this defensin is expressed constitutively in

T. villosa with leaf, stem, root, and seed showing almost similar levels of

58

high expression. The recombinant peptide (rTvD1), expressed in the

Escherichia coli expression system, exhibited potent in vitro antifungal

activity against several filamentous soil-borne fungal pathogens. The

purified peptide also showed significant inhibition of root elongation in

Arabidopsis seedlings, subsequently affecting the extension of growing

root hairs indicating that it has the potential to disturb the plant growth

and development.

Sanchez et al. (2000) investigated the possible antiviral effect on dengue

viruses of different flavonoids extracted and identified from the Mexican

plants including T. madrensis. Glabranine and 7-o-methyl-glabranine

isolated from Tephrosia sp. exerted a dose – dependent inhibitory effect

in vitro on the dengue virus.

2.11.4. Entire plant

The petroleum extract of the whole plant of I. tinctoria showed antifungal

activity against Helminthosporium sativum (Bhatnagar et al., 1961)

Efficacy of methanolic extract of whole parts of I. tinctoria against

replication of HIV-I (III B) and HIV-2 (ROD) in MT-4 cells has been

discussed by Kavimani et al. (2000). The extract exhibited an average

EC50 of 113 and 125, maximum protection of 7.5 and 9 respectively

against HIV-1and HIV-2 strains.

59

2.12. INSECTICIDAL AND PISCICIDAL ACTIVITY

Petroleum ether extract of seeds of six plants (1000, 500, 250, 125 and

62.5 ppm concentrations) were examined by Anuradha et al. (2000).

Highest mortality was observed in the seed extracts of Acaia nilotica,

Citrullus colocynthus, I. tinctoria and Maduca longifolia. Benzene

extracts of all the plants except Indigofera showed high percentage of

mortality.

Three new and two previously known flavonoids were isolated and

identified by Machocho et al. (1995) from the roots of T. emoroides

A. Rich. The new flavonoids included the flavanone 4″,5″-dihydro-5-

methoxy-5″-isopropenylfurano-[2″,3″:7,8]-flavanone, the flavone

7-hydroxy-5-methoxy-8-(3″-hydroxy)-isopent-1-eneflavone and the

pterocarpan 4′, S'-dihydro-5′-isopropenyl-8, 9-methylenedioxyfurano-

[2′,3′:2,3]-pterocarpan. The three new compounds were named

emoroidenone, emoroidone, and emoroidocarpan, respectively. The

previously known flavonoids that were isolated were the flavanone, 5-

methoxyisolonchocarpin and the flavene, hildegardtene. The flavonoids

were tested for antifeedant activity against the larvae of Chilo partellus

Swinhoe and the flavanone emoroidenone showed strong feeding

deterrent activity, percentage deterrence of 66.1% at a dose of 100

µg/disc. The other flavonoids showed little or no feeding deterrent activity

against C. partellus larvae.

60

Ignacimuthu (1998) reported insecticidal and pisicidal plants,

including Tephrosia; Botanical and their role in agriculture,

Microorganisms in insect pest managements and chemistry of plant

products in insect pest control.

Leaves of the tropical legume T. candida DC deterred feeding by

adults of the Diaprepes root weevil, Diaprepes abbreviatus (L.),

compared with leaves of Citrus macrophylla Wester, a common citrus

rootstock, or T. vogelii Hook. f. When larvae were placed in pots

containing plants of the three species for 28 days in a growth chamber,

larval survival and weight gain were significantly reduced in pots

containing plants of T. candida compared with larvae in pots with

C. macrophylla or T. vogelii. T. candida, but not T. vogelii, contains at

least one constituent that acts as an antifeedant toward adult

D. abbreviatus and as a toxicant toward larvae. No antifeedant effect of

roots of T. candida toward larvae was observed in no-choice pot tests or

in a diet incorporation bioassay. In pots, larval feeding damage to roots of

T. candida was evident. In the diet incorporation assay, 97% of larvae

survived 29 days on a diet of cellulose powder (a nutritionally inert filler)

despite losing weight. (Stephan et al., 2003)

The leaf of T. vogelii is ichthyotoxic and has been used as an

insecticide, rodenticide and anthelminthic. It has also been used as

abortifacient and to induce menses. The leaf macerate is purgative and

emetic, while the sap is used to treat diarrhoea. The leaf sap and root

61

scrapings are used as ear and tooth ache remedies respectively. Extracts

from the root have been used as a molluscicide. The plant extracts have

also been used in the treatment of tuberculosis, typhoid fever and localised

fungal infections. The biological activities are due mainly to rotenoids

isolated from the plant. (Dzenda et al., 2008)

2.13. AGRONOMY

Harvesting indigo (I. tinctoria) at little pod stage recorded 20

percentage higher biomass and 46 percent higher dye content than that at

flowering stage. Application of nitrogen increased the biomass yield, but

it decreased the dye content and dye yield. Increasing levels of

phosphorous or phosphorus recorded higher biomass dye content and dye

yield. Harvesting at little pod stage and application of 60 Kg/ha

phosphorous without nitrogen recorded highest dye content and dye yield.

(Pratibha and Korwar, 2005)

Sivapalan (1993) suggested the need to interplant shade and green

manure crops including Tephrosia in tea plantations has been critically

reassessed in holistic manner.