Simultaneous analysis of morphine‐related compounds in plasma using mixed‐mode solid phase extraction and UltraPerformance liquid chromatography–mass spectrometry

Abstract

A bioanalytical method using mixed-mode solid phase extraction and UltraPerformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was developed for the analysis of morphine, morphine-3beta-glucuronide, morphine-6beta-glucuronide, 6-acetylmorphine, morphine N-oxide, and 10-hydroxymorphine in porcine plasma. All six compounds, along with four deuterated internal standards, were simultaneously extracted using mixed-mode strong cation exchange SPE in a 96-well microElution plate format. Due to analyte instability, a neutralizing solvent was used during the elution step to minimize degradation of 6-acetylmorphine. Separation was subsequently performed in 8 minutes on a 2.1 x 100 mm, 1.8 microm C(18 )column designed for retention of extremely polar compounds using a formic acid and methanol gradient. Analytes were detected by positive electrospray ionization in multiple reaction monitoring mode using a fast-scanning triple quadrupole mass spectrometer. Recovery was 73-123% depending on the analyte, and inter-day variability was less than 6%. Linearity was determined in porcine plasma by spiking the analytes prior to SPE. Correlation coefficients were >or= 0.998, and% deviation from the actual concentrations was less than 15%. The lower limit of quantitation (LLOQ) for all compounds was between 0.1 and 0.25 ng/mL.

Simultaneous analysis of gemfibrozil, morphine, and its two active metabolites in different mouse brain structures using solid‐phase extraction with ultra‐high performance liquid chromatography and tandem mass spectrometry with a deuterated internal standard

“…The deuterated M6G-d3 was used as an IS to accurately assay the four analytes in mouse brain tissues. In comparison with the published methods, the method discussed in this study is more sensitive and rapid, and has an even lower LLOQ and more reasonable linear range of 0.05-10 ng/mL for all of the analytes with shorter analytical time (eluting morphine and its metabolites within 1.8 min) [28].…”

Forensic Science

“…A review has been published on sample preparation and separation techniques for bioanalysis of morphine and related substances . Opiates were determined in plasma using LC/ESI-MS/MS, , using SPE and quadrupole linear ion-trap LC-MS, and using mixed-mode solid-phase extraction and UPLC/MS/MS, in urine by reversed-phase UPLC/MS/MS, by liquid–liquid extraction and cation-selective exhaustive injection and sweeping micellar electrokinetic chromatography (CSEI-sweep-MEKC), by liquid–liquid extraction and GC/MS, hydrolyzed with β-glucuronidase followed by LC/MS/MS, by CE with fluorescence detection, and by SPE and HI-LC/MS/MS, and in hair by a microwave extraction and HPLC/DAD . A method for simultaneous screening and quantification was developed for opioid drugs in post-mortem blood and urine by LC/MS/MS and post-mortem blood and brain by reversed-phase LC/MS/MS .…”

Recent Trends for Enhancing the Diversity and Quality of Soybean Products

“…Multiply-charged ions such as [M + nH] n+ are often observed (Gaskell 1997). As examples, Fountain and Grumbach (2009) used negative ion electrospray mass spectrometry for the analysis of fructose, glucose, sucrose, and lactose. Taormina et al (2007) and Mauri et al (2002) used flow injection techniques with mass spectrometry.…”