Determination of Buprenorphine, Fentanyl and LSD in Whole Blood by UPLC-MS-MS

A sensitive analytical method using ultra-performance liquid chromatography-tandem mass spectrometry was developed and validated to measure buprenorphine, fentanyl, and lysergic acid diethylamide (LSD) in whole blood. Sample preparation involved liquid-liquid extraction with methyl tert-butyl ether. The method achieved detection limits of 0.28 ng/mL for buprenorphine, 0.044 ng/mL for fentanyl, and 0.0097 ng/mL for LSD. Most phospholipids were removed during extraction, and matrix effects were minimal or absent. Since September 2011, the method has been routinely used at the Norwegian Institute of Public Health for qualitative and quantitative analysis of over 400 whole blood samples, each tested in duplicate.

Study at a glance

Abstract

A sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS-MS) method has been developed and validated for the quantification of buprenorphine, fentanyl and lysergic acid diethylamide (LSD) in whole blood. Sample preparation was performed by liquid-liquid extraction (LLE) with methyl tert-butyl ether. UPLC-MS-MS analysis was performed with a mobile phase consisting of ammonium formate (pH 10.2) and methanol. Positive electrospray ionization MS-MS detection was performed with two multiple reaction monitoring transitions for each of the analytes and the deuterium labeled internal standards. Limit of detection values of buprenorphine, fentanyl and LSD were 0.28, 0.044 and 0.0097 ng/mL and limit of quantification values were 0.94, 0.14 and 0.036 ng/mL, respectively. Most phospholipids were removed during LLE. No or only minor matrix effects were observed. The method has been routinely used at the Norwegian Institute of Public Health since September 2011 for qualitative and quantitative detections of buprenorphine, fentanyl and/or LSD in more than 400 whole blood samples with two replicates per sample.