Method validation in quantitative analysis of phase I and phase II metabolites of mitragynine in human urine using liquid chromatography-tandem mass spectrometry

Mei Jin Lee, Surash Ramanathan, Sharif Mahsufi Mansor, Keng Yoon Yeong, Soo Choon Tan

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    9 Citations (Scopus)

    Abstract

    A method using solid phase extraction and liquid chromatography-tandem mass spectrometry to quantitatively detect mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine in human urine samples was developed and validated. The relevant metabolites were identified using multiple reaction monitoring in positive ionization mode using nalorphine as an internal standard. The method was validated for accuracy, precision, recovery, linearity, and lower limit of quantitation. The intra- and inter-day accuracy and precision were found in the range of 83.6–117.5% with coefficient of variation less than 13%. The percentage of recovery for mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine was within the range of 80.1–118.9%. The lower limit of quantification was 1 ng/mL for mitragynine, 2 ng/mL for 16-carboxy mitragynine, and 50 ng/mL for 9-O-demethyl mitragynine. The developed method was reproducible, high precision and accuracy with good linearity and recovery for mitragynine, 16-carboxy mitragynine, and 9-O-demethyl mitragynine in human urine.

    Original languageEnglish
    Pages (from-to)146-161
    Number of pages16
    JournalAnalytical Biochemistry
    Volume543
    DOIs
    Publication statusPublished - 15 Feb 2018

    Keywords

    • Human urine
    • Kratom
    • Liquid chromatography-tandem mass spectrometry (LC-MS/MS)
    • Metabolites
    • Solid phase extraction (SPE)
    • Validation

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