Anticancer and antioxidant Holuthuria edulis
Description
This research describes the antioxidant and anticancer activity of sea cucumber extract Holothuria edulis from Labuan Bajo. Sea cucumbers are extracted using three methods, namely maceration, soxhletation and ultrasonication. The data displayed is related to the calculation of toxicity data for the BSLT method, antioxidants and anticancer MCF-7 cells. The BSLT method toxicity calculation is as follows, the calculation of toxicity (IC50) was carried out using probit analysis. Number of dead larvae % death = ------------------------ x 100% Number of test larvae The IC50 was then calculated A blank – A sample % inhibition = -------------------- x 100% A blank The IC50 was then calculated: The absorbance of treated cells – absorbance of control media % cell death = -------------------------------------------------------------------- x 100% The absorbance of control cell – Absorbance of control media
Files
Steps to reproduce
BSLT (Brine Shrimp Lethality Test) Methode The method used in this study was a modified BSLT by . Shrimp larvae were prepared by soaking 50 mg of shrimp cysts in 300 ml of seawater in a 500 ml Erlenmeyer glass. The hatching was generated by lighting a 14-watt white lamp and aerating for 48 hours at room temperature. Five level dilutions of test solution were prepared, 1000 ppm, 500 ppm, 250 ppm, 125 ppm, and 62.5 ppm. The diluent solution was seawater. The test used a 96 wells-plate. 10 shrimp larvae were placed in each well containing the test solution with three repetitions for each concentration. Incubation was carried out at room temperature, illuminated by a 14-watt white light, for 24 hours. The surviving larvae were then counted. Negative controls were carried out using the same procedure without using the extract. The calculation of toxicity (IC50) was carried out using probit analysis. Number of dead larvae % death = -------------------------- x 100% Number of test larvae 2.5 Antioxidant Test Antioxidant was analyzed using the DPPH method. The sea cucumber extract was prepared to make solutions with concentrations of 500 µg/ml; 250 µg/ml; 125 µg/ml; 62.5 µg/ml; and 31.25 µg/ml, each concentration was added with 40 µg/ml DPPH solution. The mixture was then incubated for 30 minutes in a dark place. The absorbance was measured with three replications at a wavelength of 520 nm with a UV-Vis spectrophotometer. The reference solutions were vitamin C with concentrations of 6 µg/ml; 4.8 µg/ml; 3.6 µg/ml; 2.4 µg/ml; and 1.2 µg/ml. The IC50 was then calculated. A blank – A sample % inhibition = ------------------ x 100% A blank 2.6 Anticancer The anticancer activity was tested using the MTT assay. MCF-7 cells (ATCC HTB 22) were grown at a concentration of 5000 cells in 100 µ1 growth medium (RPM\1640 supplemented with 10% Fetal Bovine Serum (FBS) and 100 U/ml Penicillin and 100 ug/ml Streptomycin). The extract was added after the cells were 50% confluent (24 hours). The MTT test was carried out on day 3 by adding 10 µI MTT (5 mg/ml) to each well and incubating for 4 hours at 37 °C. Formazan crystals were dissolved in ethanol. The absorbance was measured at a wavelength of 595 nm. The absorbance of treated cells – absorbance of control media % cell death = ------------------------------------------------------------------- x 100% The absorbance of control cell – Absorbance of control media