Standardization and Biological activity of Calotropis gigantea

Vijay Danapur^*, Gopinath, Shravani, Annapurna G, Sinchana and Goutami Hadimani
^*Correspondence: Vijay Danapur, Department of Biotechnology, University of Davangere, Davangere, India, Tel: 9916893111, Email:

Keywords

Calotropis gigantea (L.) • Anti-diabetic • Anti-oxidant • Anti-bacterial activity • HPTLC and bio autography

Introduction

Calotropis gigantea is a minor tree or shurb belongs to family Asclepiadaceae, it is 4-10 cm tall. Its stem is straight. The plant has a bunch of the waxy flowers which are white or purple in colour. Its common name is crown flower or gaint milk weed. The leaves are oval, light green and oppositely oriented. The have cylindrical roots about 90 cms long and 2-10 cm in diameter. Stem is smooth. It is growing wide range. The flower has 5 pointed petals [1]. It is not consumed by animals. In axillary cymes with multiple flowers and a fungal tube 2-5 cm long at the base of the tomatoes pedicel makes up the floral inflorescence. The flowers have a yellow and light perfume and can be regular, lilac, bisexual, or pale pink, purple, or green [2]. They are placed laterally on the opposing sides of the nodules or at the tips of the interpetiolar peduncles in simple or infrequently mixed cymose corymbs. Numerous small, Calotropis elongated, pointed, protrusions surround each cluster. Oval flower buds appear [3].

This plant contain a chemical constituents, they are numerous glycosides, alkaloids, flavones, tannins and also calotoxin, uschrain, uscharidin and proceroside. Additionally numerous Cardenolides, flavonoids, terpenes and pregnanes are present. Calotropis, gigantea and uscharin show digitals like action on heart [4]. Calotropis has marked as anti-coagulant which is used to treat coronary thrombosis.

The leaves and Arial parts of the plant are showing the anti-diarrheal activity, anti-oxidant activity, anti-microbial activity, cytotoxic activity and wound healing activity, stem shows hepatotoxic effects [5]. Calotropis gigantea is used for the natural remedy for many diseases. The latex of these plants used in different conditions as tumors, analgesic, expectorant, piles and asthma [6].

The leaf juice of Calotropis gigantea is rubbed on the body for the body pains, especially if the joints, the fresh leaf juice are used in traditional medicine [7]. They are used as Antivenin and anti-diabetic. Dried roots used to treat asthma. The root juice in India used to reduce labour pain.

In Ayurveda the leaves of Calotropis gigantea is used in the treatment of paralysis. In Unani medicine it is used as anti-inflammatory, sedative, wound healer [8]. The root bark constitutes drug. The small doses of the root bark are diaphoretic and expectorant. It useful in the leprosy. The root bark gives relief in diarrhoea and dysentery. It also given in cough and asthma [9]. The latex is used as arrow poison in the Africa. It is an effective fish poison.

Materials and Methods

Collection

Leaf parts of Calotropis procera are collected in and around Bangalore and was identified and deposited in raw drug repository of Vriksha Vijnan Pvt Ltd, Bengaluru with the number IV/FP/VVPL/10/06/22. The plant material was screened for the presence of admixtures, foreign matters (sand, glass particles and dirt), mold or signs of decay. The plant material was later made into powder using pulverizer and the samples were stored in airtight container for further use.

Organoleptic characters

The present investigation comprises studies on both physical and sensory characteristics such as colour, sensation, taste, and texture of the species under study.

Physico-chemical evaluation

Determination of total ash: Two grams of powdered drug was incinerated in a sintered silica crucible by gradually increasing the heat to 500ºC-600ºC until the drug is free from carbon and then cooled. This kept in a dessicator for 15-20 min and weighed using Anamed electronic balance and noted down the readings.

Total ash (%)=(B-C)/(A) × 100

Where,

A=Sample weight in grams.
B=Weight of dish+contents after drying.
C=Weight of the empty dish in grams.

Determination of acid insoluble ash: Total ash obtained was boiled for 15 min in 25 ml of hydrochloric acid and filtered to collect the insoluble matter on Whatman filter paper and ignited in a sintered crucible. It was allowed to cool and then kept in a dessicator for 15 min. The residue was weighed in Anamed electronic balance and the acid soluble ash was calculated using the formula.

Acid insoluble ash (%) = (B-C/(A) × 100

Where,

A=Sample weight in grams.
B=Weight of dish+content after drying in grams.
C=Weight of empty dish in grams.

Determination of extractive values: Two grams of powdered plant material of both plants understudy were extracted with ethanol and water. Thus obtained extracts were allowed to dry to room temperature. After complete evaporation, weight, nature and colour of the extracts were recorded.

Extractive value (%)=(B-C)/(A) × 4 × 100

Where,

A=Weight of the plant material.
B=Weight of the dish+residue.
C=Weight of the empty dish.

Powder microscopy studies

The powdered plant material was soaked in 10% nitric acid overnight. The sample is washed with distilled water the following day. Slides are prepared by staining the soaked plant material with saffranine and observed under microscope and the images were captured.

Fluorescence studies

The fluorescent examination evaluating the behavior of the powder samples with various chemical regents such as, methanol, ethanol, water, concentrated HCl, H₂SO₄ and HNO₃ were observed in daylight as well as under UV radiation. Fluorescent analyses of all the plant powders were carried out according to the methods of chase and pratt.

Preliminary phytochemical tests

Extract of Illicium verum was subjected to preliminary phytochemical screening for the detection of various phytoconstituents such as alkaloids, tannins and phenolic compounds, flavonoids, steroids, saponins. This examination was done following the methods of Gibbs, Peach and Tracey.

HPTLC studies

Sample solutions were applied to the silica gel 60 F254 pre-coated TLC plates as sharp bands by means of aspire spraylin sample applicator. The spots were dried in a current of air. Chromatography was carried out in a glass chamber (aspire). The mobile phase was poured into a twin trough glass chamber whole assembly was left to equilibrate and for pre-saturation for 30 min. The plate was then developed until the solvent front had travelled at a distance of 80 mm above the base of the plate, at 20ºC and 50% relative humidity. The plate was visualized for detection by observing it under UV 254, 366 nm and after derivatization. The densitometric scan was drawn using just TLC software attached to aspire HPTLC unit.

Bio-autography method

Antioxidant and anti-diabetic activity: A solvent system of toluene, ethyl acetate and formic acid (5:4:1) was taken to develop the chromatogram. Air-dry the chromatogram for the complete removal of solvents. Later spray the chromatogram with a solution of 0.2% DPPH in methanol/ethanol observe antioxidant activity and with iodine solution for anti-diabetic activity. Chromatograms were examined in visible light.

Results and Discussion

Organolopetic evaluation

The organalopetic evaluation of the Calotropis gigantea which consists of the testing the smell, taste, texture and etc. the external colour of the flowers identified as the purple and leaves are light green in color. Texture of leaves is coriacious. The stem leaves and other parts of the plant are toxic (Table 1).