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A Novel UHPLC-MS/MS Method for the Quantification of Seven Opioids in Different Human Tissues
Alessandra Manca1, Amedeo De Nicolò1, Elisa Delia De Vivo1
1Laboratory of Clinical Pharmacology and Pharmacogenetics, Department of Medical Sciences, University of Turin, Amedeo di Savoia Hospital, Corso Svizzera, 164, 10149 Turin, Italy.
Background:
Opioids are considered the cornerstone of pain management: they show good efficacy as a first-line therapy for moderate to severe cancer pain. Since pharmacokinetic/pharmacodynamic information about the tissue-specific effect and toxicity of opioids is still scarce, their quantification in post-mortem autoptic specimens could give interesting insights.
Methods:
We describe an ultra-high-performance liquid chromatography coupled with tandem mass spectrometry method for the simultaneous quantification of methadone, morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone and fentanyl in several tissues: liver, brain, kidney, abdominal adipose tissue, lung and blood plasma. The presented method has been applied on 28 autoptic samples from different organs obtained from four deceased PLWH who used opioids for palliative care during terminal disease.
Results:
Sample preparation was based on tissue weighing, disruption, sonication with drug extraction medium and a protein precipitation protocol. The extracts were then dried, reconstituted and injected onto the LX50 QSight 220 (Perkin Elmer, Milan, Italy) system. Separation was obtained by a 7 min gradient run at 40 °C with a Kinetex Biphenyl 2.6 µm, 2.1 × 100 mm. Concerning the analyzed samples, higher opioids concentrations were observed in tissues than in plasma. Particularly, O-MOR and O-COD showed higher concentrations in kidney and liver than other tissues (>15-20 times greater) and blood plasma (>100 times greater).
Conclusions:
Results in terms of linearity, accuracy, precision, recovery and matrix effect fitted the recommendations of FDA and EMA guidelines, and the sensitivity was high enough to allow successful application on human autoptic specimens from an ethically approved clinical study, confirming its eligibility for post-mortem pharmacological/toxicological studies.
Insights
Quantifying opioids in post-mortem tissues reveals higher concentrations in organs like the kidney and liver compared to plasma. This method is crucial for post-mortem toxicological studies of opioid use.
Area of Science:
- Pharmacology
- Toxicology
- Analytical Chemistry
Background:
- Opioids are essential for managing moderate to severe cancer pain.
- Limited data exists on tissue-specific opioid pharmacokinetics, pharmacodynamics, and toxicity.
- Quantification in post-mortem samples can offer valuable insights.
Purpose of the Study:
- To develop and validate an ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method.
- To simultaneously quantify multiple opioids (methadone, morphine, oxycodone, hydrocodone, oxymorphone, hydromorphone, fentanyl) in various post-mortem tissues and plasma.
- To assess the method's suitability for post-mortem toxicological analysis.
Main Methods:
- Simultaneous quantification of seven opioids using UHPLC-MS/MS.
- Analysis of liver, brain, kidney, adipose tissue, lung, and blood plasma from 28 autoptic samples.
- Sample preparation involved tissue disruption, extraction, and protein precipitation.
Main Results:
- The UHPLC-MS/MS method demonstrated linearity, accuracy, precision, and acceptable recovery.
- Opioid concentrations were significantly higher in tissues than in plasma.
- Morphine (O-MOR) and oxycodone (O-COD) showed markedly higher concentrations in kidney and liver tissues compared to other tissues and plasma.
Conclusions:
- The validated UHPLC-MS/MS method is sensitive and reliable for opioid quantification in human post-mortem specimens.
- The findings confirm the method's eligibility for post-mortem pharmacological and toxicological investigations.
- Higher tissue-bound opioid concentrations have implications for understanding post-mortem distribution and toxicity.
