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Phytochemical Screening and Evaluation of Cytotoxicity and Thrombolytic Properties of Achyranthes Aspera Leaf Extract

Md. Jakir Hossain
Department of Genetic Engineering and Biotechnology, University of Chittagong, Chittagong-4331, Bangladesh.

Mohammad Al-Forkan
Department of Genetic Engineering and Biotechnology, University of Chittagong, Chittagong-4331, Bangladesh.

Laila Khaleda
Department of Genetic Engineering and Biotechnology, University of Chittagong, Chittagong-4331, Bangladesh.

A. M. Masudul Azad Chowdhury
Department of Genetic Engineering and Biotechnology, University of Chittagong, Chittagong-4331, Bangladesh.

Md. Arifuzzaman
Department of Biochemistry and Biotechnology, University of Science and Technology, Chittagong (USTC), Foy’s Lake, Chittagong-4202, Bangladesh.

Abstract/ Executive Summary:

The rural and marginal people of Bangladesh are deprived of modern treatment facilities and hence greatly depend on medicinal plants. Besides, the higher cost and toxicity of synthetic drugs drives scientists towards search for natural source of medication for a number of diseases. Cost-effectiveness, easy availability and fewer side effects are making the herbal medicine more popular both among rural and city people. Plants with Cytotoxic and clot lysis potential are good candidate as source of novel anti-tumor agents and thrombolytic drugs. This study aimed at screening out of phytochemical constituents and evaluation of cytotoxicity and thrombolytic potential of an important medicinal plant Achyranthes aspera methanolic leaf extract. In vitro phytochemical screening of A. aspera leaf extract carried out by qualitative tests revealed the presence of alkaloids, glycosides, cardiac glycosides, flavonoids, tannins, terpenoids, steroids and saponins while phlobatannins were absent. Cytotoxicity test of A. aspera leaf extract carried out by Brine shrimp Lethality (BSL) Bioassay showed the highest percentage of mortality (90%) in 1250 µg/ml and LC50 value was 50.12 µg/ml. Thrombolytic test showed 32.87 ± 9.42% clot lytic activity for A. aspera while positive control (streptokinase) and negative control (water) showed 81.19 ± 3.78% and 6.67 ± 2.58% clot lysis, respectively. Synergistic effect of streptokinase and A. aspera extract also produced better result (56.30 ± 6.95%) than A. aspera alone.

Keywords: Achyranthes aspera, Cytotoxic, Thrombolytic, Phytochemical screening

Location: Chittagong University Campus area.

Start Date:

End Date:

Program Area: Development of Host and Medicinal Plants

Commodity: Medicinal Plants

Objectives:

To develop improved recombinant variants of these drugs from natural sources like medicinal plants.

Methodology:

The fresh and healthy leaves of Achyranthes aspera were collected from Chittagong University Campus area. The plant was taxonomically classified and authenticated by standard taxonomical method. The fresh leaves of plant of Achyranthes aspera was washed with clean water immediately after collection. The collected leaves were chopped into small pieces, sun dried for about 5 days and grinded into coarse powder with a mechanical grinder and stored in an airtight container. 166 gm powder was macerated in 700 ml 95% pure methanol (Sigma Chemicals Co., USA) for 5 days at room temperature 25±2 °C with occasional stirring. After 5 days, methanol extract was filtered with Whatman No.1 filter paper. The extract was concentrated under reduced pressure below 50 °C through rotary vacuum evaporator. The concentrated extract was collected in a Petri dish and allowed to air dry for complete evaporation of methanol. The whole process was repeated three times and finally, 14.77gm blackish-green colored, concentrated plant extract was obtained (yield 8.9% w/w) which was kept in refrigerator at 4°C. The extract thus obtained is ready for subsequent therapeutic assessments. The cysts (eggs) of the Brine shrimp were decapsulated in 200 ppm chlorine water for 15-20 minutes. Then the cysts were washed thoroughly with fresh water till all the chlorine gas escapes and cysts were then taken in a conical hatching container and filtered (clean) seawater was added. Soaking density was 1-2 gm of cysts per litre of seawater. Moderate aeration was supplied from the bottom of the container at the rate of 10 to 20 litre of air per minute. The container was kept under illumination using a white lamp for 48 h for the eggs to hatch into shrimp larvae (nauplii). Then the nauplii were harvested in a beaker. The significance between % clot lysis by Streptokinase, herbal extract, water by means of weight difference was tested by the paired t-test analysis. Data were expressed as mean ± standard deviation.

Source of Information: Journal of Pharmacy and Biological Sciences, Volume 6, Issue 3 (May. – Jun. 2013), PP 30-38

Article Link (Online Journal): DOI:10.9790/3008-0633038

Funding Source:

Total Budget (Tk.) :

Achievement/Findings:

Assessment of cytotoxicity and thrombolytic properties of Achyranthes aspera leaf extract reveals the possibility of using this plant as anti tumor agent and it can be used in combination with other thrombolytic drugs in treating circulatory system disorders involving blood clot. This study yielded promising results with the crude extract. However, the identification, isolation and purification of active phytochemical constituents responsible for these therapeutic properties may lead to new drug development from this plant. Further in vitro and in vivo studies are recommended to harvest the benefits of natural medication from this plant.

Knowledge Management: Journal