Study On Factors Influencing Drtermination Of Anticoagulant Activity Of Whitmania Pigra Whitman By Thrombin Titration And Enzyme Extraction Craft

Background:At the earlier time,some researchers use plate method,as the tedious operation and have some error,APTT,PT also be used in determination,but as blood is very instable,so many researchers ues thrombin titration now,but it has human error,so we must further study.Objective:To analyse the factors that influence the thrombin titration for determination of anticoagulant activity of Whitmania Pigra Whitman.Method:The white porcelain was used as titration carrier instead of tube in titration,called white porcelain method for short.The effect of white porcelain method and tube method,interval time of titration and thrombin concentration on anticoagulant activity testing was studied,better titration methods were selected through comparing accuracy and stability,and human factors,such as standard titration end point states and stir frequency,were regulated by data statistical analysis.Result:1.Under the same conditions,the anticoagulant activity was more accurate and stable when we select white porcelain method.When select white porcelain method and 20 u/ml or 10 u/ml as thrombin concentration,titrate 5 μL each time,onceevery minute,the thrombin units consumption are liner with samples concentration within a certain range(0.125～0.333 g/ml),r20=0.961 and r10=0.992;the anticoagulant activity respectively was 33.08±2.64 and 31.24± 1.32 u/g,RSD20=8.0%and RSD10=4.2%;for a certain sample concentration(0.333 g/ml),the theoretical error of determination was not more than 10%and 5%.In titration,a whole piece of transparent gel was selected as standard titration end point state,it’ s more visible so would make less mis judgment;the suitable stir frequency was 10 times per minutes in titration.2.When shuxuetong was concentrated to 0.71～1.llg/ml,the results are more stable and accurate,the anticoagulant activity range is 3.83～4.23u/g.Conclusion:The improved white porcelain method is more suitable for determining anticoagulant activity of Whitmania Pigra Whitman for more stable results and accurate end point states than tube method.When we select 10 u/ml as thrombin concentration,titrate 5 μL each time,onceevery minute,the linearity,accuracy and precision were higher.