Fucoidanase activity determination method on basis of pHBH method

In this study, a new method using 4-Hydroxyben zhydrazide for the content determination of fucose reducing end was establided at frist. The detection limit of this method was 0.26 μg/mL and the quantification limit was 0.85 μg/mL. It possessed several advantages, including simple-operating, high precision and sensitivity. On base of this, we study some key factors for the fucoidanase activity detection. Then we built a fast and reliable fucoidanase activity detection method as follows: a mixture of 375 μL fucoidan solution with the concentration of 0.055% and 375 μL enzyme solution was frist reacted at 28°C for 30 minutes. After that 250 μL of pHBH was added in, the mixture was then reacted at 100°C for 5 minutes. After cooling off, the absorbance was detemined at 415 nm. Using this method, the fucoidanase activity detection limit was as low as 0.048 mU.