Studies on Standardization of Tephrosia purpurea Linn – an important Ayurvedic Drug

 

Arjun Singh²*, Ajay Kr. Meena¹, M.M. Rao¹, Sudeep Mishra², M.M. Padhi² and Ramesh Babu²

¹National Institute of Ayurvedic Pharmaceutical Research, Patiala – 47001, Punjab, (India).

²Central Council for Research in Ayurveda and Siddha, Janakpuri, Delhi-110058, (India).

*Corresponding Author E-mail: arjunccras@gmail.com

ABSTRACT:

Herbal drugs are traditionally used in various parts of the world to cure different diseases. The present study has been conducted to evaluate the quality control parameters of the hydro alcoholic extracts of Tephrosia purpurea Linn ;an important Indian medicinal plant widely used in the preparation of Ayurvedic formulations. Tephrosia purpurea Linn. is a highly branched, sub-erect herb commonly known in Sanskrit as Sharapunkha. It has been used in Indian traditional medicine for the treatment of various inflammatory disorders. The plant is used to cure diseases of the kidney, liver, spleen, heart, blood, skin eruptions, bronchitis, bilious febrile attack, boils, pimples, and bleeding piles. The dried herb is effective as a tonic, laxative, and diuretic. The roots and seeds are have insecticidal, piscicidal and vermifugal properties.

 

As there is no detailed standardisation work reported on Tephrosia purpurea Linn extract, the physicochemical parameters, phytochemical analysis, toxic heavy metals and microbial contamination analysis are carried out. The study revealed specific identities for the particular crude drug which will be useful in identification and control to adulterations of the raw drug and Ayurvedic formulations.

 

 

INTRODUCTION:

Since origin of human’s life, plants continue to play a curative and therapeutic role in preserving human health against disease and decay. The widespread use of herbal remedies and healthcare preparations, such as those described in ancient texts like the Vedas and the Bible have been traced to the occurrence of natural products with medicinal properties^1,2. In this context, India being a subtropical country is a good repository of plants that are widely used in the preparation of herbal therapies.

 

Tephrosia purpurea (Linn) Pers (Family Leguminosae) is a polymorphic, much branched sub erect perennial herb popularly known as “Sarapunkha” in Sanskrit, Sarphonk, Sarponkha in Hindi, Fish Poison, Wild Indigo, Purple Tephrosia in English and Kaattukolingi in Tamil.

 

It is a highly branched sub erect herbaceous perennial³, up to 40-80 cm in height with spreading branches; the leaves are imparipinnate, with narrow, oblanceolate leaflets; the flowers are red or purple in extra-axillary racemes, the pods are slightly curved, 3-4.5 cm long, grey, smooth and containing 5-10 seeds per pod^4,5. The plant grows abundantly in the upper Gangetic plains, and western Himalayas. The herb is commonly grown as a green manure in paddy fields in India and in tobacco and rubber plantation in other countries. It grows ubiquitously in all soils, sandy, rocky and loamy⁶. In India and South Africa, it is used as a fodder before flowering, but in Australia it is reported to cause livestock poisoning. In northern India, dry plants are collected for fuel.

 

The roots, leaves and seeds contain tephrosin, deguelin and quercetin, the roots contain isotephrosin and rotenone. In the roots and leaves 2.5% rutin is found. Purpurin, a flavonone has been isolated from the seeds, as also 8- substituted flavonoid and 3- substitued oxygenated chalcones. Octacosanol, sitosterol- C- glucopyranoside and a flavone glycoside have been isolated from the whole plant. Caffeic acid isolated from dormant seeds; s rutin, sitosterol and lupeol isolated from leaves; delphinidin chloride and cyaniding chloride isolated from flowers. Purpuritenin A and B and purpureamethide isolated from seeds. A new – hydroxychalcone – purpurenone – isolated from roots.⁷

 

All parts of the plant have tonic and laxative properties. Its aerial parts and roots are used in bronchial asthma^8,9, hepatic ailments^10,11, cutaneous toxicities¹², pain and inflammation¹³. The dried plant is deobstruent, diuretic and useful in treating bronchitis, bilious febrile attacks and obstructions of the liver, spleen and kidneys. It is also recommended as a blood purifier, in the treatment of boils and pimples and is considered a cordial treatment. In southern India, a decoction of the fruit is given for intestinal worms and a fruit extract is used to relieve bodily pains and inflammatory problems. The roots are bitter and the decoction is used as a nematicide for treatment against Toxocora canis larvae which cause a lung disease in Sri Lanka; it is also used for treating dyspepsia, colic, and chronic diarrhoea and as an antihelminthic^14-16.

 

Sarapunkha is bitter and astringent in taste, pungent is the post digestive effect and has hot potency. It has a special potency as bhedana – an accumulation breaking herb. It alleviates vata and kapha doshas. Important Ayurvedic properties of Tephrosia purpurea Linn. are Rasa  Tikta, Kashaya; Guna Lakhu, Rooksha, Teekshna; Virya Ushna; Vipaka Katu.  Medicinal properties of Tephrosia purpurea Linn. in Ayurveda, plant pacifies vitiated kapha, vata, poison, skin diseases, pruritus, jaundice, splenomegaly, filariasis, dysmenorrhoea, anaemia, fever, boils, pimple, and haemorrhoids. Manly useful part is whole plant. The important classical Ayurvedic formulations prepared from Tephrosia purpurea Linn are Sarapunkha svarasa, Sarapunkha lepa, Sarapunkha ghanavati etc.¹⁷

 

Therefore, the present paper attempts to evaluate the physicochemical parameters, phytochemical analysis, heavy metal analysis and analysis of Microbial contamination of the Tephrosia purpurea extract for identification of the drug in dry form and control the adulterants.

 

RESULT AND DISCUSSION:

Physio-chemical parameters of the Tephrosia purpurea hydro- alcoholic extract are tabulated in Table 1. The pH value of sample was acidic. Deterioration time of the plant material depends upon the amount of water present in plant material. If the water content is high, the plant material can be easily deteriorated due to fungus. The loss on drying at 105°C in sample was found to be 2.02%. Total ash value of plant material indicated the amount of minerals and earthy materials attached to the plant material. Analytical results showed total ash value content was 16.70%. The negligible amount of acid-insoluble siliceous matter present in the sample was 1.61%. The water-soluble extractive value was indicating the presence of sugar, acids and inorganic compounds, the water soluble extractive value in the sample was 85.86%. The alcohol soluble extractive value was 50.29%, it is indicated the presence of polar constituents like phenols, alkaloids, steroids, glycosides, flavonoids.¹⁸

 

Table 1. Analysis of various physico-chemical parameters of Tephrosia purpurea Linn.

S. No	Name of Parameter	Results
1.	Particle size through 40 mesh	97.63%
2.	Loss on Drying	2.02%
3.	Water Soluble Extractive	85.86%
4.	Alcohol Soluble Extractive	50.29%
5.	pH	5.30%
6.	Total Ash	16.70%
7.	Acid Insoluble Ash	1.61%
8.	Bulk Density (gm/ml)	0.75
9.	Tapped Density (gm/ml)	1.04
10.	Heavy Metals
a.	Lead	< 5 ppm
b.	Cadmium	<1 ppm
c.	Arsenic	<2 ppm
d.	Mercury	<1 ppm
11.	Solvent residue (Ethanol)	<500ppm
12.	Assay (Rutin) By HPLC Method	3.0% on d/b
13.	Microbial contamination
a.	Total Plate Count	100 Cfu/gm
b.	Yeast/Moulds	<10 Cfu/gm
c.	Enterobacteriaceae	Absent
d.	E.Coli	Absent
e.	Salmonella	Absent
f.	Staphylococcus Aureus	Absent
g.	Pseudomonnas Aeruginosa	Absent

 

The presence of Toxic heavy metals namely Arsenic, Mercury, Cadmium, Lead, phytochemical analysis and microbial contamination Bulk Density (gm/ml), Tapped Density (gm/ml) Assay (Rutin), Solvent residue were analysed in the sample and the results are shown in Table 1. The concentrations of all the heavy metals were below the Ayurvedic Pharmacopeia of India, WHO/FDA permissible limits.^18,19 The presences of microbial contamination were not detected in the samples.

 

Thin layer chromatographic technique was used to separate the chemical compounds present in the drug. Various solvent systems were checked to separate the maximum number of chemical compounds in the drug. TLC of the ethanol extract developed in the mobile phase of Toluene: Ethyl acetate: Acetic Acid :: 6 : 4 : 0.1 (Figure. 1) and observed under UV 254 nm 8 spots; under UV 366 nm showed 13 spots and after derivatization with  iodine, showed 14 spots detailed R_f values are given in Table. 2.

 

Figure 1. TLC profile of Tephrosia purpurea Linn; (Toluene: Ethyl acetate: Acetic Acid :: 6 : 4 : 0.1 v/v)

Under  λ 254 nm	Under λ 366 nm	After derivatisation with Vanillin Sulphuric acid

Table 2. TLC profile of Tephrosia purpurea Linn (Toluene: Ethyl acetate: Acetic Acid  6 : 4 : 0.1)

Sr. No	Under  λ 254 nm		Under λ 366 nm		After derivatisation with Vanillin Sulphuric acid
	Rf	Colour	Rf	Colour	Rf	Colour
1.	0.17	Green	0.08	Pale Blue	0.06	Pale Grey
2.	0.26	Green	0.14	Grey	0.10	Pale Grey
3.	0.31	Green	0.18	Blue	0.15	Pale Grey
4.	0.48	Green	0.27	Pale Blue	0.22	Purple
5.	-	-	0.31	Pale Blue	0.25	Pale yellow
6.	-	-	0.33	Pale Blue	0.29	Pale Grey
7.	-	-	0.36	Pale Blue	0.36	Pale Grey
8.	-	-	0.42	Pale Blue	0.50	Pale Grey
9.	-	-	0.47	Blue	0.55	Pale Grey
10.	-	-	0.54	Blue	0.66	Pale Grey
11.	0.66	Green	0.65	Blue	0.74	Pale Grey
12.	0.75	Green	0.74	Pale Blue	0.81	Pale Grey
13.	0.89	Green	0.95	Blue	0.87	Pale Grey
14.	0.97	Green	-	-	0.97	Pale Grey

 

 

CONCLUSION:

Various aspects of the Tephrosia purpurea hydro- alcoholic extract sample was studied and described along with, physico-chemical, Phytochemical, toxic heavy metal, microbial contaminants and TLC studies in authentification adulteration for quality control of herbal/raw drugs. Tephrosia purpurea Linn exhibits a set of diagnostic characters, which will help to identify the drug in dried condition.

 

It has been concluded from this study that estimation of heavy metals, TLC profile and microbial contamination is highly essential for raw drugs or plant parts used for the preparation of ayurvedic/ herbal compound formulations. The periodic assessment is essential for quality assurance and safer use of herbal drugs.

 

ACKNOWLEDGEMENT:

The authors are very grateful to Director General, CCRAS, New Delhi and for providing encouragement and facilities for carrying out this work. Authors acknowledge the secretarial assistance rendered by Ms. Rekha in the preparation of this paper.

 

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Received on 11.01.2011        Modified on 20.02.2011

Accepted on 28.02.2011        © AJRC All right reserved