Application Note: In-well hydrolysis using UCT Refine Ultra-Filtration Technology
15 March 2023

In this application note, UCT outlines a suggested procedure for in-well hydrolysis of urine samples using the Refine Ultra-Filtration Technology. Various biological sample types commonly contain proteins that complicate downstream analysis if not effectively removed. While sample preparation techniques, such as solid phase extraction (SPE), can successfully remove these unwanted matrix components, complex and lengthy extraction procedures are not ideal for high-throughput laboratories. With the advancement of LC-MS/MS technology, many laboratories have implemented dilute and shoot procedures in order to reduce sample preparation time, reduce consumable spending and reduce overall turnaround time. The caveat to this quick protocol is the toll that it ultimately takes on the HPLC column and mass spec source.

The Refine Ultra-Filtration Technology allows for sample precipitation and filtration to occur simultaneously in the individual column/well. A novel, hydrophobically treated, submicron frit combination facilitates the removal of sample proteins without a complicated extraction. Using the Refine Ultra-Filtration Technology reduces column backpressure while also reducing analyte suppression when compared to a standard dilute and shoot protocol.