A. T.M. Mostafa Kamal*
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203,Bangladesh.
Kazi Ashfak Ahmed Chowdhury
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Md. Masud Rana
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Azharul Islam
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Estekhar Ahmad Khan
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Md. Areeful Haque
Drug and Herbal Research Center, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, Kuala Lumpur 50300, Malaysia.
Anaytulla
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Md. Moazzam Hossen Chy
Department of Pharmacy, International Islamic University Chittagong, Chittagong-4203, Bangladesh.
Neolamarckia cadamba (Roxb); Cytotoxic activity; Thrombolytic activity; Anthelmintic activity methanol extract.
Department of Pharmacy, International Islamic University Chittagong, Bangladesh
Development of Host and Medicinal Plants
Medicinal Plants
2.1 ChemicalsLyophilised streptokinase vial (1 500 000 IU) was purchased from Square Pharmaceuticals Ltd, Bangladesh. Methanol was purchased from Merck, Germany. Normal saline solution was purchased from Beximco Infusion Ltd. Levamisole was purchased from ACI Limited, Bangladesh. All chemicals used were of analytical reagent grade.
2.2 Plant MaterialsFresh stem of C. reflexa for this study were collected from the local area of Chittagong, Bangladesh and were authenticated by Dr. Sheikh Bokhtear Uddin, Associate Professor, Department of Botany, University of Chittagong, Chittagong-4331, Bangladesh.
2.3 Preparation of Crude Extract The collected stem were dried for a period of 2 weeks under shade and ground. The ground stem(750 gm) were soaked in sufficient amount of methanol for one week at room temperature with occasional shaking and stirring. The sediments were filtered and the filtrates were dried at 40°C in a water bath. The solvent was completely removed by filtering with Whatman number-1 filter paper. The solvent was evaporated under reduced pressure at room temperature to yield semisolid. The extract was then preserved in a refrigerator till further use.
2.4 Brine Shrimp Lethality AssayThe assay was carried out according to the principle and protocol previously described by Meyer et al., with slight modifications. Here simple zoological organism (Artemia salina) was used as a convenient monitor for the screening. Dried cysts of Artemia salina were collected from an aquarium shop (Chittagong, Bangladesh) and hatched in artificial seawater (3.8% NaCl solution) for 48 h to mature shrimp called nauplii. After hatching, active naupliifree from egg covers were collected from a brighter portion of the hatching chamber and used for the assay. The test sample (extract) were prepared by dissolving them in DMSO (not more than 50 μL in 5 mL solution) plus sea water (3.8% NaCl in water) to attain concentrations of 10, 25, 50, 100, 200, 300, 500 and 800 μg/ml. A vial containing 50 μL DMSO diluted to 5 mL was used as a control. Vincristine sulphate was used as positive control. After 24 hours the number of survival of nauplii was counted and percentage of mortality was determined using the equation: % mortality = (no. of dead nauplii/ initial no. of live nauplli) x 100. Statistical method of probit analysis (Finney’s table)[35] was used to calculate LC50. Criterion of toxicity for fractions was established according to Déciga-Campos et al.[36]. LC50values > 1000 μg/mL (non-toxic), ≥ 500 ≤ 1000 μg/mL (weak toxicity) and < 500 μg/mL (toxic).2.5 Thrombolytic TestThis test was performed according to the method described by Prasad et al.[37]. In the commercially available lyophilised streptokinase vial (1 500 000 IU), 5 ml sterile distilled water was added and mixed properly. This suspension was used as a stock solution from which appropriate dilution was made. Five milliliter of venous blood was drawn from the healthy volunteers (n=10) without the history of oral contraceptive or anticoagulant therapy and was distributed (0.5 mL/tube) to each ten previously weighed sterile microcentrifuge tube and incubated at 37°C for 45 min to form the clot. After the clot formation, serum was completely removed without disturbing the clot and each tube having clot was again weighed to determine the clot weight. A volume of 100 μL of methanol extract (10 mg/ mL) was added to each micro centrifuge tube containing pre-weighed clot. As a positive control, 100 μL of streptokinase and as a negative control, 100 μL of distilled water were separately added to the control tube numbered. All the tubes were then incubated at 37°C for 90 min and observed for clot lysis. After incubation, fluid released was removed and tubes were again weighed to observe the difference in weight after clot disruption. Difference obtained in weight taken before and after clot lysis was expressed as the percentage of clot lysis.2.6 Anthelmintic AssayThe anthelmintic activity of methanolic extract of stem of C. reflexa was carried out as per the procedure of Ajaiyeoba et al.[38]with some minor modifications. The aquarium worm Tubifex tubifex were used in the present study because it has anatomical similarity and belongs to the same group of intestinal worm i.e. annelid. The worm were collected from the local market of Chittagong, average size of worms 2-2.5 cm. were taking study. The standard drug levamisole and three different concentrations of methanol extracts (2.5, 5 and 10 mg/ml) in double-distilled water were prepared freshly and used for the study of anthelmintic activity. One group was composed of water and it was considered a controlled group. The anthelmintic activity was determined at two different stage ‘time of paralysis and ‘time of death of the worms. Time for paralysis was noted when no movement of any sort could be observed except when the worms were shaken vigorously. Death was concluded when the worms lost their motility followed with fading away of their body colors. Death was also confirmed by dipping the worms in slightly warm water. The mortality of parasite was assumed to have occurred when all signs of movement had ceased.
European Journal of Medicinal Plants10(2): 1-9, 2015, Article no.EJMP.17121ISSN: 2231-0894
Journal