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The development of a high-performance liquid chromatography-tandem mass spectrometric method for simultaneous quantification of morphine, morphine-3-?-glucuronide, morphine-6-?-glucuronide, hydromorphone, and normorphine in serum.

ABSTRACT: Development and validation of a selective, robust high-performance liquid chromatography-tandem mass spectrometric (HPLC/MS-MS) method for the quantification of morphine, morphine-3-?-glucuronide, morphine-6-?-glucuronide, hydromorphone, and normorphine in human serum.Drug-free human serum samples spiked with morphine, morphine-3-?-glucuronide, morphine-6-?-glucuronide, hydromorphone, and normorphine were prepared by protein precipitation using methanol containing the internal standards. Samples were injected onto a Thermo Scientific AccuCore PFP column for chromatographic separation. Detection was achieved using a Thermo Scientific TSQ Vantage mass spectrometer. Assay validation followed the new Clinical and Laboratory Standards Institute (CLSI) C62-A guidelines.The analytical measuring range for all analytes was determined to be 5 to 1000 ng/mL. Intra- and inter-assay precision for three quality control levels were ? 7.0% and ? 13.5%, respectively. Carryover, stability, linearity, matrix effects, extraction and processing efficiency and method comparison characteristics were acceptable relative to the CLSI C62 guidelines.The validation of this HPLC-MS/MS method demonstrated a robust and rapid assay for the quantification of morphine, morphine-3-?-glucuronide, morphine-6-?-glucuronide, hydromorphone, and normorphine.

SUBMITTER: Sartori D

PROVIDER: S-EPMC4679425 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature

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