Phytochemical and Ethnopharmacological Evaluations of Methanolic Extracts of Seed Coat of Manilkara Zapota : In-vitro & In-vivo Approaches.

The plant Manilkara zapota , commonly known as chicle or sapodilla, is a slender, slow-growing, pyramidal evergreen tree that is native from southern Mexico to Costa Rica. This plant is spread all over the world including American tropics, Spain, Brazil and Bolivia, Pakistan, Srilanka etc. Phytochemical & Ethnopharmacological Evaluations of Methanolic Extracts of Seed Coat of Manilkara Zapota: In-vitro and In-vivo approaches have been investigated. It was used in ancient system of medicine in many countries but not to great extent. The fruit is used as reducing infections, in viral disease, bacterial infections, in tooth cavity, blood cell formation. Moreover, it also used in swelling, as a pain relief agent. It is suggested for the insomnia and panic disorder. More over Sapodilla fruit is full of vitamin A, C and B complex. So, its beneficial for good health, eye and tooth. In the Phytochemical screening the methanolic extract of Manilkara zapota seed coat showed the presence of alkaloid, flavonoid, carbohydrate, glycoside, tannin. The methanolic extract of Manilkara zapota seed coat showed 56.23% potential activity of HRBC membrane stabilization (anti-inflammatory activity) compared to standard diclofenac. The methanolic extract of seed coat also produced 60.44% antioxidant activity compared to standard ascorbic acid. So, the methanolic extract of Manilkara zapota seed coat exhibited the moderate free radical scavenging activity (anti-oxidant activity). Methanolic extract of Manilkara zapota seed coat has a good analgesic and neuropharmacological activity. Therefore, Manilkara zapota seed coat has a guiding competence for the improvement of innovative superior efficacy drug in future.


Introduction
The numerous plants have therapeutic exploits and wield valuable the pharmacological upshots on the animal bodies are typically assigned as the "Medicinal Plants". At the same time as there is the no perceptible morphological asset in the medicinal plants producing with them, as far as this they have more than a few special qualities that generate them medicinally so significant [1]. Over last few decades, exercise of herbal drugs has been accentuated due to their effortless availability, therapeutic impending, slightest side effects in addition to minimum cost. At current nearly 80% of the world population relies on plant-based drugs for their health care need [2]. Presently, phytoconstituents are playing pivotal role for development of novel compounds, which might be crucial for maintaining a healthy society. [1] The human civilization has been maintaining an intimate relationship with the plants from time immemorial [3].
They depend on plants and other natural sources for their wellbeing and survival. Various plants still available in the nature are yet to be explored for their medicinal potential. Uses of computational chemistry to discover, enhance, or study drugs and related often used to predict the conformation of the small molecule and to model conformational changes in the biological target that may occur when the small molecule binds to it [5]. This research work was carried out to evaluate the extract value, analgesic activity, antioxidant activity, anti-inflammatory and neuropharmacological activity of Manilkara zapota.

Materials and methods
The plant material named as Manilkara zapota based on its medicinal uses. Using standard taxonomical methods, the plants were collected from Chittagong division. It was then separated and cleaned from impurities ( Figure A & Table 1). The dried pure extracts were weighed separately with the help of electronic balance and the yield was determined by using the following formula [6].

Activity [7]
The antioxidant potentialof the methanolic extract was determined on the basis of their scavenging activity of the stable 1, 1-diphenyl-2-picryl hydrazyl (DPPH) free radical. DPPH is a stable free radical containing an odd electron in its structure and usually utilized for detection of the radical scavenging activity in chemical analysis. The aliquot of the different concentrations of the extract was added to 3ml of a 0.004% EtOH solution of DPPH. Absorbance at517 nm was determined after 30 minutes, and IC50 (Inhibitory concentration 50%) was determined. IC50 value denotes the concentration of sample required to scavenge 50% of the DPPH free radicals. The formula used for % inhibition ratio is % inhibition = (Control -Sample / Control) × 100

Analgesic Test Preparation of Test Dose
The extracts were suspended in the vehicle. Various strengths were prepared from a stock solution 40mg/ml. The solutions were prepared freshly, and were administered orally. The Acetic acid induced writhing test method is given below [8]: b) The reference group (Group II) received Diclofenac Na (50 mg/ kg) c) The control group received normal saline (10 mL/kg). p. o. 30 minutes after treatment was carried out, each of the mice was administered with acetic acid (0.7% v/v in saline, 10 mL/kg, i.p.) to create pain sensation.
d) Five minutes after the administration of acetic acid, the numbers of writhes were counted over a period of 15 min. The response of was compared with those of the control group.
e) % inhibition of writhing in comparison to control group was taken as an index of analgesia and was calculated using the following formula: Where, W c = average number of writhing reflexes in the control group W t = average number of writhing in the test groups.

Results and Discussion
Antioxidant Activity

Anti-Inflammatory Activity [9]
( Figure 2) It was observed from the present study that methanolic extract of has anti-inflammatory activity and activity increases with increasing extract concentration. The Maximum inhibition was found at 2000 µg/ml extract concentration which showed 56.23% inhibition compared to standard. The activity increases with increasing extract concentration.

Open Field Method
The methanolic extract of Manilkara zapota was subjected to

Rota Rod Method
The methanolic extract of Manilkara zapota was subjected to screening for muscle coordination activity by Rota rod method.

EPM Method
The methanolic extract of Manilkara zapota was subjected to

Hole Cross Method
The methanolic extract of Manilkara zapota was subjected to screening for anti-depressant activity by hole cross method. The test was performed by taking methanolic extract at doses of 200 mg/kg and 400 mg/kg body weight. The result was found moderate ( Figure 9).

Conclusion
The results from the experiment confirmed that these may be potential candidate for the treatment of antioxidamt, anti-inflam- Hence Manilkara zapota has a leading capacity for the development of new good efficacy drugs in future.