Evaluation of phytochemical, anthelmintic activity and antidiabetic activity-melochia corchorifolia ethanol extract

Melochia corchorifolia belongs to the Sterculiaceae family and is a common weed distributed throughout tropical and subtropical regions in many countries. Melochia corchorifolia is a medicinal plant which is used as a traditional folk medicine for the treatment of the various diseases. Several studies have reported antibacterial, antifungal, cytotoxicity effects and anti-cancer activity of water and methanol extracts of the leaf. The dried leaves were powdered and extracted with ethanol solvent through the Soxhlet apparatus. The phytochemical analysis was done using standard techniques on the ethanolic extract. The analysis showed the presence of various phytoconstituents such as flavonoids, alkaloids, steroids, terpenoids, glycosides, tannins, phenols, saponins and anthraquinones. Besides, the research was aimed at assessing the Anthelmintic behaviour of ethanol extract of Melochia corchorifolia leaves against Eisenia fetida (Indian earthworms)  the result shown that 300 mg/ml proved to be very active by paralyzing and killing the earthworms in a shorter time and followed by  Antidiabetic activity in  inhibition assay for α-amylase activity was evaluated using Melochia corchorifolia leaves showed maximum inhibition of the enzyme with the highest value of 85.0% seen at 100mg/ml. Our results reveal that ethanol extract of the leaf of Melochia corchorifolia acquires potent bioactive phytocompounds that might be developed into novel Anthelmintic and Antidiabetic activities.


INTRODUCTION
The plants used as an important medicinal source in human culture and civilization in the Egyptian, Assyrian, Chinese and Indian valleys know that the elders and men of that time used related medicinal plants for various diseases (Ajaib et al., 2010). Plant-derived medicinal products are popular for their health, easy availability and low cost. Herbal medicinal products may include whole parts of a plant or mostly prepared from roots, leaves, bark, seeds and plant lowers (Amar et al., 2012). The plants of medicinal value lie in components of bioactive substances that induce particular physiological exploit to the human body. The bioactive phytochemical components are tannins, saponins, alkaloids, terpenoids, lavonoids, essential oils, and phenolic compounds. Many more such natural compounds formed the basis for modern medicinal drugs (Pieroni, 2000). The genus Melochia corchorifolia is the family of Sterculiaceae has extraordinary economic importance and medicinal properties. Melochia corchorifolia, is also called as choco-late weed, is a typical weed plant available in the farmland. It was noted as an excellent source of iber (Pullaiah, 2014;Mamatha et al., 2018). Traditionally, leaves are very much useful for many medicinal remedies. It has been used to diminish ulcers, abdominal swelling, headache, and chest pain. The pharmacological activities of plant leaves showed various activities like as Anthelmintic activity (Patel et al., 2011), Hepatoprotective and Antioxidant ability, Antibacterial activity, Diuretic, Anti-Cancer activity and Antiurolithic activity on this plant (Alhakmani et al., 2013a;Wickramaratne et al., 2016).

Plant material
The plant M. corchorifolia L., are collected from the Reserve forest area, Vandalur, Thiruporur, Kanchipuram Dt. The plant was taxonomically identi ied and authenticated by Prof.P.Jayaraman, Ph.D., and Director. Institute of Herbal Botany, Plant Anatomy research centre Chennai. The leaves were washed and shade dried for 15 days to eliminate chlorophyll content. The dried leaves were inely powdered by using a mechanical blender. The 100g of dried powder leaves extracted by soxhlet method by using solvent viz. ethanol. After extraction, the extract was dried by distilling the solvents in a rotary vacuum evaporator.

Phytochemical analysis
The ethanol extract of was Melochia chorchorifolia subjected to preliminary phytochemical test using standard protocol (V and M, 2017).

Total Phenol Content Determination
The phenol content ethanol leaves crude extract of Melochia chorchorifolia was evaluated by using Folin-Ciocalteu's assay, and gallic acid was standard solution (Alhakmani et al., 2013b). The standard procedure was followed by plant extract 10.0 ml and 2.0 ml distilled water then mixed with 0.6 ml Folin-Ciocalteu's reagent (FCR) added after 10 minutes and about 1.6 ml freshly prepared 20% sodium carbonate was added. The absorbance of the sample measured against the blank at 320 nm by using a spectrophotometer (Chandra et al., 2014).

Total Flavonoid Content Determination
The lavonoid content Melochia chorchorifolia leaves ethanol extract was assessed by the Aluminum chloride method and quercetin as a standard (Soares et al., 2015). The test sample of 2ml, 5 ml water, 0.30 ml, 5% Sodium nitrite and then 0.3 ml 10% Aluminum chloride added, after that 5 minutes incubated at room temperature. The absorbance of the sample was measured against the blank at about 520 nm by using a spectrophotometer.

Anthelmintic test of ethanol plant leaves extract of Melochia chorchorifolia
The adult Indian earthworms (Eisenia fetida) were collected from the vermicomposting site of Hindustan College of Arts and Science, Padur, Tamil Nadu, India. For this experiment, adult earthworms of approximately 4 cm long and 0.2-0.3 cm broad were used. Due to its physiological and anatomical similarities to human intestinal roundworm parasites, this organism was chosen for Anthelmintic behavior (Heidari et al., 2005). The earthworms washed in normal saline before assay.
The Anthelmintic assay accomplished with minor modi ications by using standard methodology. The test sample plant extract prepared on the concentrations (100, 200, 300 mg/ml) added with distilled water then boiled for 10 minutes and mixture was iltered, collected supernatant was utilized for the Anthelmintic tests. The Adult Indian earthworms (E. fetida) of almost equal size were placed each in the petri dish having leaf extract. Albendazole used as a reference standard in the concentrations 25 mg/ml, 50 mg/ml and normal saline water as the control. The observations were made paralysis, and the death of worms was recorded.
The 500µl plant extract solution and 500µl sodium phosphate buffer containing αamylase solution (0.5mg / ml) were incubated at 25 • C for 10min. After pre-incubation, in 0.02 M sodium phosphate buffer (pH 6.9 with 0.006 M sodium chloride) 1 per cent starch solution for 500 µl has been applied to each tube at each appropriate interval (Hullatti and Telagari, 2015). At 25 • C for µl, the reaction mixture was incubated and then incubated for 5 minutes in a boiling water bath, and cooled to room temperature. About 10ml of distilled water was applied to the reaction solution to dilute the solution, and the absorbance was estimated at 540 nm (Kazeem et al., 2013). Percentage inhibition is calculated as Abs control

Phytochemical screening
The phytochemical tests Melochia chorchorifolia     leaves, which shows abundantly in positive for alkaloids, polypeptide, Phenols, Flavonoids, and Alkaloids and negative for coumarins the results are tabulated (Table 1).
The screening of plant part revealed that the amount of total phenol and lavonoid content in which leaves indicates the presence of high phenolic content which may be due to presence of phenol, lavonoid and possess antioxidant activity (Table 2 and Figure 1). Evaluated the antibacterial activity of different extracts of Melochia chorchorifolia on eight bacterial strains by using cup plate method (Rao et al., 2002).

Anthelmintic activity
At 100 mg/ml concentration Melochia chorchorifolia leaves extract-treated earthworms had taken less time for paralysis at 60 min and death of earthworms at 80 min. At 300 mg/ml concentration Melochia chorchorifolia leaves extract proved to be more potent by recording the time of paralysis of earthworms at 17 minutes and time of death of earthworms at 25 min. (Table 3 and Figure 2).

Inhibition assay for α-amylase activity (DNSA)
The DNSA assay analysis resulted are showed in (Figure 3 and Table 4). Melochia corchorifolia demonstrated high inhibition of the enzyme at all concentrations with the highest value of 85.0% reported at 100mg / ml plant extract concentration.
The phytochemical screening Melochia corchorifolia leaves ethanol extract revealed the secondary metabolites like triterpenes, carbohydrates, glycosides, lavonoids, and alkaloids, etc. The antimicrobial activity possessions on plant groups may be due to phytochemical chemical compounds in the plants (Cowan, 1999).The above phytochemical constituents are the primary source for the plant's therapeutic properties and synthesis of new medicine today (Harborne and In Phytochemical methods , 1984).
Similarly, noticed Anthelmintic activity of Melochia corchorifolia against Pheritima Posthuma (Indian worm) (Palaksha et al., 2013). Present research is evident that plant extracts of Melochia corchorifolia have shown Anthelmintic activity against Eisenia Fetida at the short span of paralytic and death time showed ethanolic extracts of Punica granatum, Mangifera indica has an in-vitro inhibitory effect on α-amylase (Prashanth et al., 2001). Present study shows Melochia corchorifolia has an excellent inhibitory effect up to 85% on α-amylase, which decreases insulin resistance in diabetic.

CONCLUSION
The main bene its of utilizing plants are that they never exhibit the frequently linked deleterious side effects of other allopathic medications. Based on the above results we conclude that Melochia corchorifolia leaves ethanol extract showed secondary metabolites in phytochemical screening and activity effect on Anthelmintic and antidiabetic. Future researches isolated the compound through spectral studies from the leaves of the medicinal plant and check the ef icacy of the compound through biological activity.