Electroanalytic, photometric or fluorometric methods may provide information about the presence of proteolysis in a related sample, but they cannot accurately identify which protease belongs to the proteolytic activity. In other words, they cannot distinguish the proteases according to the differences in their activities. Previous studies on the chymotrypsin and trypsin have shown that the activities of proteases can be distinguished from each other by fluorescence lifetime distributions. In this study, it is aimed to show the sensitivity of the distributional model on the proteolytic activities of the two proteases. For this purpose, the proteolytic activities have been reduced by making covalent conjugation with polyacrylic acid (PAAc) and the effects of the low activities were examined on Bovine Serum Albumin (BSA) which excimer emission character was incorporated into its structure by modification with N-(1-pyrenyl)maleimide. The time-resolved spectrofluorometer was used to collect fluorescence decay data at λ(excimer) = 464 nm, which were analyzed by using Exponential Series Method (ESM) to obtain the changes of lifetime distributions. The results showed a significant decline in the activities. Despite the very low activities, the changes of fluorescence lifetime distributions exhibited remarkable specific differences that proved the sensitivity of ESM analysis.

电分析，光度法或荧光法可提供有关样品中蛋白水解存在的信息，但它们不能准确地识别哪种蛋白酶属于蛋白水解活性。换句话说，它们不能根据其活性的差异来区分蛋白酶。以前对胰凝乳蛋白酶和胰蛋白酶的研究表明，蛋白酶的活性可以通过荧光寿命分布来区分。在这项研究中，旨在显示分布模型对两种蛋白酶蛋白水解活性的敏感性。以此目的，通过与聚丙烯酸（PAAc）共价偶联降低了蛋白水解活性，并研究了低活性对牛血清白蛋白（BSA）的影响，该蛋白通过用N-（1-吡啶基）修饰而被纳入了准分子发射特性）马来酰亚胺。使用时间分辨荧光分光光度计收集在λ（准分子）= 464 nm处的荧光衰减数据，并使用指数级数法（ESM）对其进行分析以获得寿命分布的变化。结果显示活动明显减少。尽管活性很低，但荧光寿命分布的变化仍表现出明显的特定差异，证明了ESM分析的灵敏度。使用时间分辨荧光分光光度计收集λ（准分子）= 464 nm处的荧光衰减数据，并使用指数级数法（ESM）对其进行分析以获得寿命分布的变化。结果显示活动明显减少。尽管活性很低，但荧光寿命分布的变化仍表现出明显的特定差异，证明了ESM分析的灵敏度。使用时间分辨荧光分光光度计收集在λ（准分子）= 464 nm处的荧光衰减数据，并使用指数级数法（ESM）对其进行分析以获得寿命分布的变化。结果显示活动明显减少。尽管活性很低，但荧光寿命分布的变化仍表现出明显的特定差异，证明了ESM分析的灵敏度。