Quantitative Opiate, Opioid, and Benzodiazepine Testing by Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)

Fatalities due to opioids and other controlled substances continue to increase year over year and maintain an epidemic level nationally. CDC data indicates that drug overdoses killed more than 100,000 Americans in 2021 with synthetic opioids being the main driver of the national crisis (Wide-ranging online data for epidemiologic research (WONDER). Multiple Cause of Death 1999-2020. Centers for Disease Control and Prevention, National Center on Health Statistics. http://wonder.cdc.gov . Accessed 28 Feb 2023, 2021). Response to this crisis includes the development of laboratory testing to support the evolving drug trends. Urine drug testing provides clinicians with objective information to assist in making treatment decisions by identifying the presence of potential drugs of abuse and assessing patient compliance to a controlled substance prescription regimen. In this chapter, we describe a rapid LC-MS/MS method for the simultaneous quantitation of 21 opiate, opioid, and benzodiazepine drugs in less than 4 min. The method requires 100 uL of urine and includes enzymatic hydrolysis of glucuronide conjugates using β-glucuronidase to provide total drug measurement. Quantitation is accomplished using multiple reaction monitoring (MRM), a seven-point calibration curve, and deuterated internal standards. This method provides a robust and reliable means to measure commonly prescribed opioid (including natural, semi-synthetic, and synthetic) and benzodiazepine drugs in urine.