Routine testing for drugs of abuse (DOA) in urine is commonly performed by many clinical, forensic, and pain management laboratories. The method of testing varies but can often provide unwanted load stress for day-to-day operation in labs. Most seek simplified yet reliable and robust modes of sample preparation and analysis. Many laboratories also desire automation for simpler, more reliable methods of extraction that reduce labour costs. Hydrolysis was conducted using the Kura Biotec B-One enzyme and the manufacturer’s suggested conditions on the Biotage® ExtraheraTM Automated Extraction System. A panel of 10 conjugated metabolites of common DOAs were hydrolysed and extracted by supported liquid extraction using ISOLUTE® SLE+. Hydrolysis efficiencies were calculated to assess method functionality. The B-One enzyme from Kura Biotec is an enhanced recombinant enzyme that allows for a short, room temperature hydrolysis. The enzyme comes as a mix with a buffer and is added to the urine sample. It was recommended that the enzyme be added in a 1:1 ratio to the urine sample by the manufacturer. The Biotage® ExtraheraTM Automated Extraction System provides automation of the extraction process for supported liquid extraction, solid phase extraction, phospholipid depletion, protein precipitation, or dual mode extraction methods. The full extraction process from sample pretreatment and loading onto an extraction plate/cartridge to washing of the plate/cartridge and elution can be performed on the system without the need for analyst intervention.
Literature number: P221.V.1