Method Establishment And Verification Of Tenecteplase Activity Detection By Chromogenic Substrate Method

As an activator of plasminogen,Tenecteplase,which is a clinical drug to treat myocardial infarction,can activate human plasminogen to plasmin to dissolve thrombus.At present,Bubble Method is approved in China as the activity detection method,which mainly has problems of large fluctuation,low accuracy and precision.In order to obtain a more stable and reliable method,Chromogenic Substrate Method is establishedS-2251,the Chromogenic Substrate,reacts with Tenecteplase,and the release of this reaction process can be detected at the wavelength of 405 nm.Based on this principle,a method for detecting the relative binding activity of Tenecteplase by Chromogenic Substrate Method was established,meanwhile the optimum experimental conditions were obtained.The diluent was selected mixed solution contain Tris-HCl,sodium chloride and human albumin.The initial concentration was 55000 IU/ml,as well as the proportion dilution was used.The detection time is determined to be 30 min.Method Verification to validate the feasibility of Tenecteplase activity detected by Chromogenic Substrate Method was investigated.The conclusion shows as follow.Specificity result is fine that the chromogenic reaction occurred to the sample containing Tenecteplase.The relative standard devition of six independent repeated experiments by one person was less than 10%,and with two people when one performed three times it was less than 15%,which all met the standards,shows degree of precision meets the requirement.The correlation coefficient R of the four-parameter fitting curve were greater than 0.99,which met the standard requirements.The recovery rate of five different starting concentrations was between 85% and 115%,that shows good accuracy.In conclusion,this method is suitable for the detection of relative binding activity of Tenecteplase.Compared with the bubble method,Chromogenic Substrate Method is better in linear and repeatability.