In this report, a completely automated, 96 well plate format “Prep-and-Shoot” workfl ow including enzymatic hydrolysis, dilution and injection is described. A GERSTEL MultiPurpose Sampler (MPS) coupled to an AB SCIEX QTRAP® 4500 LC/MS/MS system was used for a fast enzymatic hydrolysis process (15 minutes), dilution and injection of urine samples. The procedure was applied to the analysis of multiple drug classes (e.g., opiates, opioids, benzodiazepines, muscle relaxants, hallucinogens) in urine.
This automated workflow employed an ultra-pure β-Glucuronidase enzyme yielding hydrolysis effi ciencies of glucuronide conjugates above 80 % for the analytes tested. The methodology developed allowed the reproducible injection and analysis of over 960 samples on the same analytical column, with % RSDs ≤ 10 %. Moreover, the combined automation of urine hydrolysis, injection and analysis allowed the system to process more than 200 samples in a day.
