This application note describes a rapid and broadly applicable SPE protocol and UPLC-MS/MS method for analyzing a comprehensive panel of compounds common to forensic toxicology screens. The unique, water-wettable nature of the Oasis sorbent allowed us to eliminate the common conditioning and equilibration steps without any loss in recovery or reproducibility for any of the 38 compounds in this testing panel. This property also enables the entire hydrolysis step to be conducted within the wells of the Oasis MCX μElution Plate, eliminating the time-consuming and error-prone transfer steps. Combining this with the consolidation of two wash steps into a single one further facilitates the reduction of a six-step extraction process into only three steps. Though this procedure remains slightly more timeconsuming than sample dilution, it nonetheless can be completed in 30 minutes. Moreover, it offers the additional added benefits of increased sensitivity, reduced matrix interferences, increased analytical column lifetimes, and reduced risk of ion source-fouling.
This method was designed for the analysis of enzymatically hydrolyzed samples. Yet the use of the ACQUITY UPLC BEH Phenyl Column also enables the resolution and analysis of morphine-3-glucuronide and morphine-6-glucuronide, allowing the method’s use for direct analysis without hydrolysis. It also allows monitoring of the metabolites of these glucuronides. Because these compounds have been shown to be difficult to fully hydrolyze using beta glucuronidase,2 monitoring their presence can be an important factor in ensuring complete conversions to the free drugs.
This method enables the rapid extraction and analysis of a large panel of drugs for forensic toxicology screening. When combined with the chromatography of the ACQUITY UPLC BEH Phenyl Column, it provides a rapid, specific method with the sensitivity and reproducibility required to accurately screen for this panel of compounds.