Survey of Enzymatic Activities Assayed by the HPLC Method

Survey of Enzymatic Activities Assayed by the HPLC Method 

The use of HPLC to assay enzymatic activities has continued to grow in popularity since publication of the first edition of this book in 1987. Papers describing methods of assay for approximately 150 different enzymes were reviewed in preparation for writing this chapter. 

There are several reasons for the development of HPLC-based methods for assaying enzymatic activities. First is the desirability of accurately assessing enzymatic activity in preparations as near the biological state as possible. This often requires the use of turbid preparations, and the product to be measured must be separated from other components of the reaction mixture. However, the use of crude preparations often carries the price of needing to correct for secondary reactions. HPLC allows secondary reactions to also be measured. 

Second, the resolving power of HPLC greatly exceeds that of other separation methods. An enzymatic assay that was developed to use thin-layer or paper chromatography may readily be improved by switching to HPLC. Furthermore, HPLC is automated. 

the hazards of handling and disposing of radioisotopes have made it desirable to find alternative methods of assay. When the assay involves fluorescent or intensely absorbing substrates or products, the sensitivity of HPLC-based assays rivals that of radiochemical assays. 

---

SURVEY OF ENZYMATIC ACTIVITIES ASSAYED BY THE HPLC METHOD... 

---