A preliminary study of different concentration of crude proteinfrom green buffalo leech, Hirudinea manillensis on time of blood clotting

Malaysian local leech Hirudinea menillensis contain a protein compound with anti-coagulant agent which proved to be able to inhibit blood from clotting quickly (Tan & Liu, 2002; Huang et al, 2006a, 2006b; Li el al., 2006). In this study the protein quantification and anti-coagulant agent activity of crude protein from leech extract are reported. Conventional method was used to prepare the crude protein from leech extract. The results
shows 60 µl of crude protein shows the higher amount of protein concentration with a value of 1.855 mg/ml compared to 50 µl, 40 µl, 30 µl, 20 µl, 10 µl, 0 µl of crude protein which was 1.797 mg/ml, 1.722 mg/ml, L598 mg/ml, 1.52 mg/ml, 1.517 mg/ml, and 1.338 mg/ml, respectively. The time of blood clotting and time of blood did not contract was recorded by using drop method. Variable treatment from crude protein from leech extract was tested at fresh rabbit blood. For each treatment, three replicate were applied, T6 showed the longest time of blood clotting and clot did not contract with mean and standard deviation value of 5.44±0.13 and 9.45±0.05 minutes compare to all other treatment followed by
5.35±0.17 and 9.10±0.011 minutes for T5, 5.27±0.075 and 8.49±0.011 minutes for T4, 5.22±0.028 and 8.49±0.011 minutes for T3, 4.47±0.017 and 8.47±0.025 minutes for T2,
3.41±0.005 and 8.44±0.011 minutes for T1, and, 1.00±0.00 to 2.00±0.00 minutes for blank, respectively. These result indicated crude protein extract from leeches contain an anti-coagulant agent that inhibit blood from clotting and T6 have the most optimum anti-coagulant agent activity compared to other treatments because higher amount of protein concentration contain a large quantity of anti-coagulant agent.