Outpacing Emerging Drug Threats: Validation of ToxBox Kits That Automate LC-MS/MS Analyses

The Centers for Disease Control and Prevention (CDC) recently began addressing critical informational gaps associated with the emergence of novel psychoactive substances (NPSs) through the formation of the Overdose Data to Action (OD2A) biosurveillance program. This program uses public health laboratories (PHLs) to report drug trends in nonfatal and fatal overdose cases, but meeting the OD2A program's goal of reporting use trends in near real-time is a challenge for resource-limited PHLs. New technology that automates NPS testing will help PHLs meet this goal and sustain OD2A testing. This study uses commercially available test kits capable of automating clinical and forensic testing platforms to validate a single liquid chromatography tandem mass spectrometry (LC-MS/MS) analytical procedure for opioids, barbiturates, benzodiazepines, cannabinoids, stimulants, and several other drug classes. Kits incorporated Suspended-State and Just-Go technologies to stabilize NIST-traceable standards and enzymatic buffers at precise concentrations in a format that does not require sample cleanup or dilution prior to LC-MS/MS analysis. OD2A test kits were activated at 37 °C prior to the addition of 10 μL sample aliquots, and hydrolysis of conjugated urinary metabolites was complete after a 90 min incubation inside the autosampler held at 37 °C. Samples were injected directly on the column for LC-MS/MS analysis and method validation. Accuracy, precision, measurement uncertainty, calibration models, reportable range, sensitivity, specificity, carryover, interference, ion suppression/enhancement, and analyte stability met the testing requirements established for toxicology laboratories. Suitability studies using fortified human urine demonstrated that the automated LC-MS/MS method validated for OD2A biosurveillance produces highly accurate and precise results. The LC-MS/MS method was also successfully transferred and validated in an independent state public health laboratory to demonstrate how this new technology can be used to support a national biosurveillance program. This new approach will enable PHLs to provide actionable data in a timely manner.