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目的:比较不同提取方法对宽体金线蛭提取物抗凝活性的影响,确定抗凝活性物质的较佳提取方法。方法:应用凝血酶滴定法、APTT试剂盒法、PT试剂盒法等3种检测方法比较干品冷提、干品热提、干品脱脂后热提及鲜品冷提等提取物中抗凝活性的差异。结果:与空白对照组比较,各种提取方法所得提取物均具有明显的抗凝活性,各种检测方法所得结果一致。其中,水蛭鲜品和干品脱脂后提取物中抗凝活性较强。结论:水蛭提取物的抗凝活性强弱与原料的处理和提取方法有关。水蛭干品脱脂有利于提高提取物的抗凝活性。凝血酶滴定法可作为提取物抗凝活性的初步检测方法,其结果稳定,易于操作。APTT、PT试剂盒法可作为抗凝活性进一步检测的方法。
OBJECTIVE: To compare the effects of different extraction methods on the anticoagulant activity of extracts of Hirudo dabryanus and determine the best extraction method of anticoagulant actives. Methods: Three methods of thrombin titration, APTT method and PT method were used to compare anticoagulant in cold extract of dry product, thermal extract of dry product, Activity differences. Results: Compared with the blank control group, the extracts obtained by various extraction methods had obvious anticoagulant activity, and the results obtained by various detection methods were consistent. Among them, the fresh lecithin and dried skim extract after strong anticoagulant activity. Conclusion: The anticoagulant activity of leeches extract is related to the processing and extraction of raw materials. Leech dry skim products help to improve the anticoagulant activity of the extract. Thrombin titration can be used as a preliminary detection of anticoagulant activity of the extract, the result is stable and easy to operate. APTT, PT kit method can be used as a further measure of anticoagulant activity.