부산대학교 도서관

Background: Aberrant drug test results are a significant occurrence among patients undergoing chronic opioid therapy. Monitoring medication compliance and drug abuse is common practice in chronic pain management clinics, as it can increase patient prescription drug compliance and reduce illicit drug abuse. There is also interest in directly quantifying certain glucuronide metabolites to better inform clinicians about drug metabolism in their patients. However, most current pain management panels do not directly quantitate glucuronide metabolites due to several technical difficulties associated with their quantification. Despite widely available immunoassays, chromatography-mass spectrometry-based methods are considered the gold standard for therapeutic drug monitoring due to their high sensitivities and specificities. Herein, we describe the validation of a fast, convenient ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) assay to simultaneously quantify 35 prescription drugs, drugs of abuse, and related glucuronides and other metabolites, in human urine by single sample injection. Methods: A total of 35 analytes were chosen for quantification or semi-quantification based on clinician feedback. Analytes consisted of prescription and illicit opioids, benzodiazepines, and stimulants, including parent compounds and glucuronidated and non-glucuronidated metabolites. Urine samples were prepared by diluting with water and spiking it with deuterated internal standards. No enzymatic hydrolysis, analyte extraction, or sample purification were required. Analytes were separated by reverse-phase UPLC and quantified by positive-mode electrospray ionization and collision-induced dissociation mass spectrometry. Assay validation followed Food and Drug Administration (FDA) bioanalytical guidelines. Results: The assay was validated in accordance with current FDA recommendations and the anticipated testing clinical context. Total analytical run time was 5 min. All analytes demonstrated acceptable linearity (r2 >.99), accuracy (DEV <15%), precision (CV <15%), and clinically relevant analytical ranges (1-2,000 ng/mL, depending on analyte). Stability and matrix effects were also assessed for each analyte. Assay performance for each analyte was compared to either reference laboratory or previously validated in-house methods. Conclusions: A convenient and fast UPLC-MS/MS assay for simultaneously monitoring 35 clinically relevant analytes was developed and validated for use in chronic pain management at a tertiary medical center. Advantages of this assay include (1) simple sample preparation, which avoids extraction and hydrolysis steps, (2) higher-throughput multi-analyte detection including glucuronide metabolite quantification, and (3) a potential reduction in send-out confirmation testing by avoiding the use of immunoassays. [ABSTRACT FROM AUTHOR]