Study On Heavy Metal Residue And Anticoagulant Activity Of Leech

ObjectionMedical leeches are Whitmaniapigra Whitman Hirudonipponia Whitman、 Whitmaniaacranulate Whitman.Currently on the market with the largest amount of leech Whitmania Pigra Whitman.In this study, the leeches were 90 days farming, medical leeches processed into periodic sampling, using ICP-MS technique and its pharmacopoeia method for determination of heavy metals and residual antithrombin activity evaluation, the study of animal leech growth environment, taken into the feed, breeding medical leeches in time and heavy metal residues, potency of antithrombin relationship.Through market segmentation medical leech later testing, research and distribution of heavy metals in different parts of the law of antithrombin activity of medical leeches.Method1.Aquaculture Experimental section:This section includes different environments breeding experiments and feed difference breeding experiments. Leeches aquaculture seedlings are from Shandong Weishan YouBo medical leech breeding base, age about 30 days, the initial weight 2-3g. Environmental differences breeding experiments carried out in eight glass aquarium, depending on the concentrations of farmed soil, artificially contaminated Cr, Cd, Pb, Cu, and simulate different environments leech farm animals, breeding cycle for 90 days, the sampling period for 10 days, low temperature drying dry. Differences in culture experiments feed 72 polyester plastic flasks, divided snail, blood powder, no diet group, the breeding cycle for 90 days, the sampling period of 15 days, low temperature drying. Of processed medical leech heavy metals content and antithrombin potency testing.2. Different Parts experiment:medicine market collected eight leech medical leech in accordance with all body about one-third of the length is divided into head, middle, tail and all four groups received a total of 32 samples, respectively, samples of heavy metals testing and evaluation of anti-thrombin activity.3. The heavy metal content detection method:using inductively coupled plasma mass spectrometry (ICP-MS) technology for all medical leech samples before and after the experiment soil six heavy metals blood meal feed (chromium, arsenic, cadmium, mercury, lead, copper) content in testing.4. antithrombin potency evaluation methods:Method for determination of the activity of the 2015 edition of "Chinese Pharmacopoeia" in the medical leech antithrombin for all medical leech antithrombin titer samples were evaluated.Results1. medical leech class antithrombin titer assay using 2015 edition of "Chinese Pharmacopoeia" in the titration, laboratory cultured seven constituencies, only in the end of the experiment group and the high concentration of pollution-free diet group did not reach the required standard. In different medical parts of the experiment, the head of the group Status best compliance rate was 87.5%.2. medical leech heavy metals in ICP-MS detection using quantitative analysis techniques, including regression equation Cr element is y= 1.2116x-0.1772, R2= 0.9999; regression equation As elements of y= 0.0764x-0.001, R2= 1; regression equation Cd elements is y= 0.1198x-0.0049, R2 = 0.9999; regression equation Hg element is y= 0.8916x+0.0006, R2= 0.9999; Pb in the regression equation is y= 6.6566x+0.0734, R2=.1; regression equation Cu element is y= 0.2482x+0.0486, R2= 0.9999, and precision instrument test results show that three of the six detecting elements parallel detection of all values within a predetermined range of reference materials. ICP-MS technique can trace medical leech heavy metal elements for effective testing.ConclusionMedical leech anti-thrombin activity and heavy metal content of no direct precise relationship medical leech heavy metals in the body does not directly affect the effectiveness of the medical leech; the medical leech in mercury, chromium, copper content there is a strong positive relationship, and lead and cadmium content also has a positive correlation with each other. Medical leech substances antithrombin potency with the extension of animal breeding leech time increases; medical leech breeding environment have anticoagulant activity of high concentrations of pollution have a greater negative impact.Medical leeches in chromium, arsenic, lead element content will increase with time breeding and growth, the copper content was reduced after an initial increase, cadmium, mercury content and aquaculture No significant relationship; breeding higher levels of chromium in the soil, cadmium, lead elements will have a greater impact in medical leech chromium, cadmium, lead elements of the residual amount of medical leeches copper content and copper content in the culture environment was no significant relationship. As a blood meal to feed farmed animals leech, leech overall quality of their medicines better.Medical leech heavy metals in different parts of the relationship between the amount of residual elements of the head 《all《Central≈tail, and the distribution of Cd obvious differences; the relationship between antithrombin potency substances for head 》tail≈all》Central, different comprehensive quality parts order of the head 》tail》central.Cultural environment of soil, feed, water, medical leeches are other heavy metal elements in the source.InnovationThe first case of heavy metal residues and medical leech antithrombin activity linked study the relationship between them.For the first time in a laboratory simulation of animal breeding leech growth environment for a long time, and by setting different concentrations of pollution of the environment, feeding feed difference, medical leech antithrombin active substances, heavy metal elements accumulation in the case of external factors, the growth time Relationship.For the first time by the medical leech into different parts of the study of its anti-thrombin substances, heavy metals content distribution.