Development and validation of a bioanalytical method for the simultaneous determination of heroin, its main metabolites, naloxone and naltrexone by LC–MS/MS in human plasma samples: Application to a clinical trial of oral administration of a heroin/naloxone formulation
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Author/sMoreno-Vicente, Raquel; Fernandez-Nieva, Zuriñe; Navarro-Alvarez, Arantza; Gascon-Crespi, Irene; Farre-Albaladejo, Magi; [et al.]
A bioanalytical method using high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed and validated for simultaneous quantification of heroin, its main metabolites and naloxone. In addition, naltrexone was detected qualitatively. This method was used to analyse human plasma samples from a clinical trial after oral administration of a heroin/naloxone formulation in healthy volunteers. O-methylcodeine was used as an internal standard. Samples were kept in an ice-bath during their processing to minimize the degradation of heroin. A short methodology based on protein precipitation with methanol was used for sample preparation. After protein precipitation, only the addition of a formic acid solution was needed to elute heroin, 6-monoacetylmorphine, morphine, naloxone and naltrexone. Morphine metabolites were evaporated to dryness and reconstituted in a formic acid solution. Chromatographic separation was achieved at 35 C on an X-Bridge Phenyl column (150 x 4.6 ...