Simultaneous analysis of morphine-related compounds in plasma using mixed-mode solid phase extraction and UltraPerformance liquid chromatography-mass spectrometry.
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.
Fountain
Kenneth J
KJ
Waters Corporation, Milford, MA 01757, USA. kenneth_fountain@waters.com
Yin
Zhe
Z
Diehl
Diane M
DM
D000818:Animals; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D009020:Morphine / Q000737:chemistry* / Q000378:metabolism; D009022:Morphine Derivatives / Q000032:analysis*; D009294:Narcotics / Q000737:chemistry* / Q000378:metabolism; D010949:Plasma / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D052616:Solid Phase Extraction / Q000295:instrumentation / Q000379:methods*; D013552:Swine
D009022:Morphine Derivatives; D009294:Narcotics; D009020:Morphine