The object of the present invention is a new method for fluorometrically assaying the activity of fat-degrading enzymes, such as lipases and phospholipases in samples containing said enzyme, such as in serum. According to the method the enzyme containing sample is reacted with a substrate containing acyl- or alkylglycerols or -phosphoglycerols having at least one fluorescent group, such as a pyrene group. The compounds may in addition contain fluorescence quenching groups. The enzyme hydrolyzes the substrate thus giving rise to changes in the fluorescence intensity during the enzyme reaction and the changes in the intensity are measured at a specific emission wavelength of the fluorescent group employed. The rate of change of the invensity is proportional to the enzyme activity in the sample.
The invention concerns also the new compounds containing fluorescent groups to be used in the method.
本发明的目的是一种用荧光测定含
脂肪酶和
磷脂酶的样品(如血清)中脂肪降解酶活性的新方法。根据该方法,含酶样品与含有至少一个荧光基团(如
芘基)的酰基或烷基
甘油或
磷脂甘油的底物反应。此外,这些化合物还可能含有荧光淬灭基团。酶
水解底物,从而在酶反应过程中引起荧光强度的变化,强度的变化是在所使用的荧光基团的特定发射波长下测量的。本发明的强度变化率与样品中的酶活性成正比。
本发明还涉及将用于该方法的含有荧光基团的新化合物。