BRINE SHRIM LETHALITY BIOASSAY OF METHANOLIC CRUDE EXTRACTS OF TERMINALIA ARJUNA

BRINE SHRIM LETHALITY BIOASSAY OF METHANOLIC CRUDE EXTRACTS OF TERMINALIA ARJUNA

Md. Anaet Hossen Maruf ¹ and Mohammad Musarraf Hussain *²

Department of Pharmacy, Noakhali Science and Technology University ¹, Sonapur, Noakhali-3802, Bangladesh

Department of Pharmacy, Jagannath University ², Dhaka-1100, Bangladesh

ABSTRACT

The aim of this work is to determine the cytotoxic activities of Terminalia arjuna (Family: Combretaceae) by brine shrimp lethality bioassay. The leaves of Terminalia arjuna were extracted with methanol and the methanolic extract was fractionated into three fractions by using organic solvents like n-hexane, ethyl acetate and chloroform. The n-hexane, ethyl acetate and chloroform soluble fractions of methanolic crude extract were screened for antitumor properties using brine shrimp lethality bioassay. A reputed cytotoxic agent vincristine sulphate was used as a positive control. From the results of the brine shrimp lethality bioassay it could be well predicted that n- hexane, ethyl acetate and chloroform soluble fractions of Terminalia arjuna leaves possess cytotoxic principles (LC₅₀ 1.29 µg/mL, 1.5 µg/mL and 1.23 µg/mL respectively) comparison with positive control, vincristine sulphate (LC₅₀ 0.563 µg/mL).

Brine shrimp lethality bioassay

INTRODUCTION: Terminali arjuna (Local name: Arjun, Family: Combretaceae) is a tree  about 60-80 feet in height, large, evergreen with a spreading crown and drooping branches. It is simple, grey and smooth on external surface. The bark is thick, soft and of red color from inside, glabrous and often in equilateral. Leaves are oblong, 4-6 inch long, 2-3 inch wide, sub opposite.

The fruits are 1-1.5 inch in diameter with 5-7 longitudinal lobes, glabrous with five to seven wings, woody, fibrous, drupe, often notched near the top and marked with oblique upward curving striations ¹. Lot of medicinal agents and been isolated from Terminali arjuna like olenane triterpine glycoside ², ursane triterpine ²¹, cytotoxic agents ¹⁷ and free radical scavenger ²¹.

Terminali arjuna had been used as a potent medicinal agent in healing activity ³, gastro-protective ⁴, ulcer protective ⁵, cardiovascular disorder6, anti-inflammatory ⁷, Immunomodulatory ⁷, anti-nociceptive ⁷, myocardial fibrisosis8, inhibitory effects ⁹, phytomedicinal activity ¹⁰, cardioprotective ¹¹, anti-oxidant ¹⁵, adrinomycin-induced DNA damage ¹⁶, antigenotoxic properties ¹⁸ and protective effects ¹⁹.

METHODS AND MATERIALS:

Collection and proper identification of the plant sample: Plant sample (Leaves) of Terminalia arjuna was collected from Bhola in April, 2010.

Preparation of Mother Solution: Powdered material of the plant (450 g) was taken in a clean desiccator (5 L) and soaked with methanol (1.5 L).

with its content was sealed by foil and kept for a period of 20 days accompanying occasional shaking and stirring. The whole mixture was then filtered through filter paper and the filtrate thus obtained was concentrated at open air dry.

Extraction of the Plant Material: Methanolic extract of Terminalia arjuna leaves (5 g) was triturated with methanol (100 mL) containing distilled water (10 mL). This is the mother solution, which was partitioned off successively in three solvents (n-hexane, ethyl acetate and chloroform) of different polarity by modified Kupchan partitioning method and thus three extractives like n-hexane (1.1 g), chloroform (0.04 g) and ethyl acetate (0.6 g) were obtained.

FIGURE 1: SCHEMATIC DIAGRAM OF A MODIFIED KUPCHAN PARTITIONING METHOD

RESULT AND DISCUSSION: The brine shrimp lethality bioassay is a rapid, inexpensive and simple testing method of cytotoxic and anti-tumor properties determination of plant extract ¹¹. Following the procedure of Meyer, 12, 14 the cytotoxicity of methanolic crude extracts (n-hexane, ethyl acetate and chloroform soluble fractions) were determined and the LC50 values of n-hexane, ethyl acetate and chloroform soluble fraction found to be 1.29 mg/mL, 1.5mg/mL and 1.23 mg/mL respectively comparison with the positive control, vincristine sulphate (Table 1).

TABLE 1: LC₅₀ VALUES OF METHANOLIC CRUDE EXTRACTS OF TERMINALIA ARJUNA

TABLE 2: EFFECT OF n-HEXANE, ETHYL ACETATE & CHLOROFORM SOLUBLE FRACTION ON SHRIMP NAUPLII

FIGURE 2: EFFECT OF VINCRISTINE SULPHATE ON BRINE SHRIMP NAUPLII DEPENDS ON LOG C

FIGURE 3: EFFECT OF n-HEXANE SOLUBLE FRACTION ON BRINE SHRIMP NAUPLII

FIGURE 4: EFFECT OF ETHYL ACETATE SOLUBLE FRACTION ON BRINE SHRIMP NAUPLII

FIGURE 5: EFFECT OF CHLOROFORM SOLUBLE FRACTION ON BRINE SHRIMP NAUPLII

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    How to cite this article:

    Hossen MA and Hussain MM: Brine Shrim Lethality Bioassay of Methanolic Crude Extracts of Terminalia arjuna. Int J Pharm Sci Res. 3(11); 4297-4300.

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