BRINE SHRIM LETHALITY BIOASSAY OF METHANOLIC CRUDE EXTRACTS OF TERMINALIA ARJUNAHTML Full Text
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
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).
Terminalia arjuna, Combretaceae, Methanolic extracts, Cytotoxic activities,
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
|Methanolic crude extracts (soluble fractions)||LC50 (mg/mL)||Regression equation||R2|
|n-hexane||1.29||Y = 45.08x – 8.235||0.918|
|Ethyl acetate||1.5||Y = 43.67x – 15.47||0.897|
|Chloroform||1.23||Y = 44.88x – 4.984||0.902|
|Vincristine sulphate (positive control)||0.563||Y = 30.056x + 56.016||0.9168|
TABLE 2: EFFECT OF n-HEXANE, ETHYL ACETATE & CHLOROFORM SOLUBLE FRACTION ON SHRIMP NAUPLII
|Log C||Mortality (%)||LC50 (mg/mL)||Vincristine sulfate|
|n-hexane||Ethyl acetate||Chloroform||n-hexane||Ethyl acetate||CF|
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.
Md. Anaet Hossen Maruf and Mohammad Musarraf Hussain *
Assistant Professor, Department of Pharmacy, Jagannath University, Dhaka-1100 Bangladesh
02 April, 2012
21 October, 2012
29 October, 2012