1. Quantitative determination of R/S-methadone in human serum using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry: A method for routine use.
- Author
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Hansen M, Havnen H, Andreassen TN, Spigset O, and Hegstad S
- Subjects
- Humans, Stereoisomerism, Limit of Detection, Reproducibility of Results, Chromatography, High Pressure Liquid methods, Analgesics, Opioid blood, Substance Abuse Detection methods, Tandem Mass Spectrometry methods, Methadone blood, Chromatography, Supercritical Fluid methods
- Abstract
Methadone has two enantiomers, which exhibit differences in pharmacological effects, with R-methadone being the active and S-methadone the inactive enantiomer. A robust, simple and rapid method for chiral separation of the two enantiomers in serum samples using ultra-high performance supercritical fluid chromatography-tandem mass spectrometry (UHPSFC-MSMS) has been developed and validated. Enantiomeric separation was achieved using a Chiralpak IH-3 column with a mobile phase consisting of CO
2 and 30mM ammonium acetate in methanol/water (98/2, v/v). Runtime was 4 minutes. Sample preparation was semi-automated using a Hamilton ML Star robot with protein precipitation, and phospholipid removal was carried out using a Waters OSTRO™ 96-well plate. The calibration range was 50.0-1,500 nM for each enantiomer. The between-assay relative standard deviations were in the range of 1.2-3.6%. Matrix effects ranged from 99% to 115% corrected with internal standard. The method has been implemented in our laboratory and has proven to be a robust and reliable method for determining the ratio of R/S-methadone in authentic patient samples., (© 2024 The Authors. Drug Testing and Analysis published by John Wiley & Sons Ltd.)- Published
- 2024
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