IN VITRO CYTOTOXIC EFFECT OF METHANOLIC CRUDE EXTRACTS OF OCIMUM SANCTUM

IN VITRO CYTOTOXIC EFFECT OF METHANOLIC CRUDE EXTRACTS OF OCIMUM SANCTUM

Aparna Debnath 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 research is to observe the cytotoxic activities of methanolic crude extracts of Ocimum sanctum (Family: Lamiaceae). The leaves of Ocimum sanctum was extracted with organic solvent (methanol) and the extracts were fractionated by using solvent-solvent partition. The n-hexane, ethyl acetate, and chloroform soluble fractions of methanolic crude extract of Ocimum sanctum were screened for cytotoxic activity 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 was well predicted that n-hexane, ethyl acetate, and chloroform soluble fractions of methanolic crude extracts possess cytotoxic principles (LC₅₀ 4.36 mg/mL, LC₅₀ 5.37 mg/mL and LC₅₀ 10.00 mg/mL respectively) comparison with positive control, vincristine sulphate (LC₅₀ 0.563 mg/mL).

Cytotoxic effects

INTRODUCTION: Ocimum sanctum (Local name: Tulsi, Family: Lamiaceae) is an aromatic plant which is native throughout the world tropics and widespread as a cultivated plant and an escaped weed. It is an erect, much branched subshrub 30-60 cm tall with hairy stems and simple opposite green leaves that are strongly scented. Leaves have petioles, and are ovate, up to 5 cm long, usually slightly toothed and flowers are purplish in elongate racemes in close whorls ¹.

Ocimum sanctum has been used as a potent traditional medicinal agent in the antifungal ², acetylcholin-esterage and improve cognition condition ³, immuno-modulatory ^{4, 13}, antiplasmodial ^{5, 25}, radioprotective ^{6, 14}, antidiabetic ^{7, 15}, genoprotective 8, hepatoprotective ⁹, ameliorative ¹⁰, larvicidal ¹⁸, antioxidant ^{19, 20, 22}, antityrosinase activity ²⁰, reproductive ²¹, lipid lowering ²², antihyperlipidemic and cardioprotective ²³ and anti-dengue activity ²⁴.

METHODS AND MATERIAL:

Collection and identification of the plant: Plant sample of Ocimum sanctum was collected from Noakhali Science and Technology University campus in January 2010 and selected for cytotoxic activities.

Plant material preparation: The leaves of the plant were collected in fresh condition. It was sun-dried to make it suitable for grinding purpose. The coarse powder was then stored in air-tight container with marking for identification and kept in cool, dark and dry place for future use.

Extraction of plant material: The powdered leaves (700 g) were soaked in methanol (2.8 L) in May, 2010 in a desicator, through occasional shaking and stirring for 25 days. The extract was then filtered through filter-cloth. The filtrate was kept to dry in fresh and clean air to afford a greenish mass of methanolic crude extarct. The methanolic crude extract was fractionate according to Modified Kupchan partitioning method (figure 2) into n-hexane (0.04 g), ethyl acetate (0.6 g) and chloroform (0.06)  soluble fractions.

FIGURE 1: SCHEMATIC DIAGRAM OF THE CRUDE EXTRACTS OF OCIMUM SANCTUM

FIGURE 2: SCHEMATIC DIAGRAM OF A MODIFIED KUPCHAN PARTITIONING METHOD

RESULT AND DISCUSSION: The Brine Shrimp test (BST) represents a rapid, inexpensive and simple bioassay for testing cytotoxic and anti-tumor properties of plant extracts ¹¹. Following the procedure of Meyer ¹² and Persoone ¹⁷, the cytotoxic effects of the 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 fractions found to be 4.365 mg/mL 5.370 mg/mL and 10.00 mg/mL respectively (Table 1) compared with positive control, vincristine sulphate (0.563 mg/mL). From the test it was observed that cytotoxic effects exhibited by the methanolic extract (n-hexane, ethyl acetate and chloroform soluble fractions). The effects of n-hexane, ethyl acetate, chloroform soluble fractions and vincristine sulphate on shrimp nauplii are shown in the table 2 and figure 3, 4, 5 and 6 respectively.

TABLE 1: LC₅₀ VALUES OF METHANOLIC CRUDE EXTRACTS OF OCIMUM SANCTUM

TABLE 2: EFFECT OF N-HEXANE, ETHYL ACETATE, CHLOROFORM SOLUBLE FRACTIONS AND VINCRISTINE SULPHATE ON BRINE SHRIMP NAUPLII

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

FIGURE 6: EFFECT OF VINCRISTINE SULPHATE ON BRINE SHRIMP NAUPLII

CONCLUSION: The present research indicates that the crude extracts of Ocimum sanctum has got intense in-vitro cytotoxic effect and may have potential use in traditional medicine.

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    How to cite this article:

    Debnath A and Hussain MM: In vitro cytotoxic effect of Methanolic crude extracts of Ocimum sanctum. Int J Pharm Sci Res 2013; 4(3); 1159-1163.

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