Nuzhat Mahbub
Department of Pharmacy, East West University, Dhaka, Bangladesh
Most. Nasrin Akter
Department of Pharmacy, BRAC University, Dhaka, Bangladesh
Tushar Ahmed Shishir
Department of Pharmacy, BRAC University, Dhaka, Bangladesh
SM Saiful Islam
Department of Pharmacy, BRAC University, Dhaka, Bangladesh
Nafisa Nawar
Department of Pharmacy, BRAC University, Dhaka, Bangladesh
Sterculia coccinea, Antioxidant, Antimicrobial, Antidiarrheal, Hypoglycemic, Thrombolytic activity
National Herbarium Bangladesh, Mirpur, Dhaka
Development of Host and Medicinal Plants
Medicinal Plants
The leaf part of Sterculia coccinea plant was collected in May 2017 from the Chittagong hill tract. After collection, the National Herbarium Bangladesh (NHB), Mirpur, and Dhaka authenticated the plant material and provided a plant identification number, which was 47695. At first, the leaves part was washed with fresh water to remove the unwanted dust particles and plant scrap. After that, the cleaned leaves were dried under the sun for a day. Then the leaves were again dried for 1 hour at 30-400C in a hot air oven. By using a high capacity grinding machine, the dry and crusty leaves were ground. After that, at a normal ambient temperature (22-250C) around 900 g of ground powder was soaked in 2.5 L of methanol for a period of 2 days with occasional stirring. With the help of a cotton filter (pore size: 110mm) filtration was done and a rotary evaporator was used at 100 rpm at 300C to evaporate the maximum amount of solvent. For vaporizing the solvent completely from the extract, the leaf extract was kept under a laminar airflow cabinet. Moreover, it was used to avoid any possibility of microbial growth in the extract while drying. Finally, 22.4 g of plant leaf extract was obtained and kept in a dry and cool place and proper labeling was done. After that, this extract was used to conduct antioxidant, brine shrimp lethality assay, thrombolytic, antidiabetic, antimicrobial and hypoglycemic studies. The phenols were oxidized by Folin-Ciocalteu in an ionic phenolic solution. When the solution became yellow to dark blue, it is understood that the oxidation has been completed. After that, this color changed mixture measured at 760 nm in UV spectrophotometer. Finally, the value of the absorbance plotted in the gallic acid calibration curve and data were evaluated as gallic acid equivalents (GAE). Aluminum chloride was used to determine the total amount of flavonoids. Firstly, 0.5 ml of plant extract was made the final volume of 1 ml for reaction medium (MeOH/H?O/CH?COOH=14:5:1) which was then mixed with Aluminum chloride reagent (4 ml, 133 mg of Al Cl × 6 H2O and 400 mg of CH? COO Na dissolved in 100 ml H2O). After 5 minutes, the absorbance was measured at 430 nm. Based on the calibration curve, total flavonoid content was calculated and it was expressed as gram equivalents. The antioxidant activity of Sterculia coccinea was determined by performing DPPH free radical scavenging assay. To run this assay, different concentrations of plant extracts were mixed with 2, 2-diphenyl-1-picrylhydrazyl (DPHH) solution. In methanol or aqueous solution, free radicals were generated due to delocalization of the free electrons and a deep purple colored solution is produced. Then absorbance of different concentration solutions was measured at 517 nm in UV spectrophotometer. The decreasing value of DPHH at 517 nm is directly proportional to the radical scavenging activity. Percentage of inhibition of DPHH free radical (1%) was calculated by using the following equation: (1%) = (Absorbance of blank - Absorbance of the sample)/Absorbance of blank ×100; 50% of inhibition of the extract concentration was calculated from the graph and the percentage of inhibition was plotted against extract concentration. In this assay, Artemia salina shrimp was used. Its offspring was hatched in replicated seawater to cultivate nauplii. Here, the calculated amount of dimethyl-sulfoxide (DMSO) was added with the sample and desired concentration of the sample was prepared by dilution. The counted nauplii were placed in vials that contained approximately 5 mL simulated seawater with a visual inspection. With the help of micropipette, various concentrations of samples were added to tubes. Here, vincristine sulfate was used as standard. The sample containing tubes were then placed in a dry place for 24 hours at room temperature. At the last, after 24 hours, the survived nauplii were counted. Percentage (%) of mortality was calculated by using the following equation: Percentage of mortality= (Number of nauplii taken - Number of nauplii alive)/ Number of nauplii taken ×100. 50% of lethal concentration of extract concentration was calculated from the graph plotted percentage of mortality against concentration. The normal blood flow to the cells and tissues can be hampered due to thrombus as it blocks the blood vessel which can lead to a lack of blood and oxygen. There are some thrombolytic medications like utokinase, clopifogrel, and streptokinase that remove this thrombus and cells and tissues remain in normal conditions. For this assay, fresh human blood was collected. Then, they were taken in three different pre-weighed sterile microbes and incubated for 45 minutes at 37°C. The upper fluid was entirely dispensed from all micro- tube lines when the clot has appeared. As standard streptokinase was used and as a negative control distilled, water was used. 100 microliter of plant extract was taken in each tube and incubated for 90 minutes at 37°C. Next, liquid that was released from the clot was removed and the tubes were weighed again to observe the weight difference when the clot disruption occurred. Percentage of clot lysis was calculated by the following equation: (%) of clot lysis = (released clot weighted) / (clot weight after clot disruption) ×100. In recent years, different studies are developing antimicrobial agents to fight antibiotics resistance from different sources and the highest concentration has given to screen and evaluate the antimicrobial activity. By using disc diffusion assay method, antimicrobial activity of Sterculia coccinea was evaluated. E. coli bacteria (gram negative) and Bacillus Subtilis bacteria (gram-positive) were used in this study. Mular Hinton agar (MHA) was used as media in this assay. Firstly, every petri dish was autoclaved for sterilization and 20 ml of MHA was poured in every petri dish. After that, the plates were kept for a time being to be settled. With the help of a cotton swab, the nutrient broth of bacterial strains was incubated in MHA. A small disc of filter paper was made by using a paper punch machine and then different concentrations of plant extract (200 mg/mL and 400 mg/mL) were used to swallow that filter paper. When the discs become dry, they were transferred to the Petri dishes and kept in the incubator for 24 hours at 370C. After 24 hours the zone of inhibition was calculated and for keeping the contamination limited, the whole experiment was done under laminar flow.
Journal of Medicinal Plants Studies 2019; 7(4): 113-117
Journal