Development and validation of a radioreceptor assay for the determination of morphine and its active metabolites in serum

Autor: J.P. Franke, Rainer Bischoff, Donald R. A. Uges, Marieke P.H. Kroeze, Katharina Krämer, Lutea A.A. de Jong
Přispěvatelé: Pharmaceutical Analysis, Analytical Biochemistry
Jazyk: angličtina
Rok vydání: 2005
Předmět:
Metabolite
Clinical Biochemistry
Receptors
Opioid
mu
Pharmaceutical Science
morphine-6-beta-glucuronide
Pharmacology
Ligands
OPIATE RECEPTOR
Analytical Chemistry
chemistry.chemical_compound
Radioligand Assay
Opioid receptor
Drug Discovery
Freezing
HUMAN PLASMA
Spectroscopy
validation
NEWBORN-INFANTS
morphine
quantitative radioreceptor assay
Analgesics
Opioid
SOLID-PHASE EXTRACTION
morphine-3-glucuronide
μ-opioid receptor
Algorithms
medicine.drug
medicine.drug_class
therapeutic drug monitoring
Analgesic
In Vitro Techniques
BINDING-SITES
OPIOID RECEPTOR
PRETERM INFANTS
medicine
Animals
Active metabolite
Morphine-3-glucuronide
Morphine Derivatives
Chromatography
Reproducibility of Results
MASS-SPECTROMETRY
Enkephalin
Ala(2)-MePhe(4)-Gly(5)
FENTANYL RECEPTOR ASSAY
PERFORMANCE LIQUID-CHROMATOGRAPHY
Neostriatum
Opioid
chemistry
mu-opioid receptor
Cattle
Indicators and Reagents
serum
Zdroj: Journal of Pharmaceutical and Biomedical Analysis, 39(5), 964-971. ELSEVIER SCIENCE BV
ISSN: 1873-264X
DOI: 10.1016/j.jpba.2005.04.049
Popis: This article describes the development and validation of a radioreceptor assay for the determination of morphine and morphine-6-beta-glucuronide (M6G) in serum. The assay is based on competitive inhibition of the mu-opioid-selective radiolabeled ligand [3H]-DAMGO by opioid ligands (e.g. M6G) for binding to the striatal opioid receptor. The assay has been validated according to the Washington Conference Report on Analytical Method Validation. The radioreceptor assay can be performed in serum without prior pre-treatment of the sample. Direct addition of the sample results in no significant loss in maximal binding sites, and therefore, no loss in sensitivity. The assay proves to be selective for a multitude of opioid agonists and antagonists (e.g. morphine IC50 = 4.1 nM and M6G IC50 = 12.8 nM). Moreover, morphine-3-glucuronide (M3G) displays a low affinity (IC50 = 1100 nM) for the mu-opioid receptor and according to the literature demonstrates no analgesic activity. This makes discrimination, in relation to the analgesic effect, of the two metabolites of morphine possible. The assay is fast (assay time4h, analysis 5 min/sample), easy and the sensitivity (limit of detection (LOD) = 1.6 nM M6G-equivalents) is such that very potent agonists, like morphine and M6G, can be measured at the desired serum levels. The assay is accurate (18%), but precision is limited if measured over several days (35%). The assay is most accurate and precise if measured over a range from 3.5 to 40 nM M6G-equivalents. Based on the limited inter-assay precision, we propose to use this receptor assay mainly as a screening tool for neonates treated with morphine.
Databáze: OpenAIRE
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