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

Raquel Moreno-Vicente, Zuriñe Fernández-Nieva, Arantza Navarro, Irene Gascón-Crespí, Magí Farré-Albaladejo, Manuela Igartua, Rosa María Hernández, José Luis Pedraz

Research output: Contribution to journalArticleResearchpeer-review

10 Citations (Scopus)

Abstract

© 2015. 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. ×. 4.6. mm, 5. μm) using a gradient elution with a mobile phase of ammonium formate buffer at pH 3.0 and formic acid in acetonitrile. The run time was 8. min. The analytes were monitored using a triple quadrupole mass spectrometer with positive electrospray ionization (ESI+) in multiple reaction monitoring (MRM) mode. The method was found to be linear in a concentration range of 10-2000. ng/mL for M3G and 10-1000. ng/mL for the rest of compounds. Quality controls showed accurate values between -3.6% and 4.0% and intra- and inter-day precisions were below 11.5% for all analytes. The overall recoveries were approximately 100% for all analytes including the internal standard. A rapid, specific, precise and simple method was developed for the determination of heroin, its metabolites, naloxone and naltrexone in human plasma. This method was successfully applied to a clinical trial in 12 healthy volunteers.
Original languageEnglish
Pages (from-to)105-112
JournalJournal of Pharmaceutical and Biomedical Analysis
Volume114
DOIs
Publication statusPublished - 1 Oct 2015

Keywords

  • Heroin
  • LC-MS/MS
  • Pharmacokinetic
  • Plasma
  • Validation

Fingerprint Dive into the research topics of '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'. Together they form a unique fingerprint.

Cite this