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Synthesis and bioanalytical evaluation of morphine‐3‐O‐sulfate and morphine‐6‐O‐sulfate in human urine and plasma using LC‐MS/MS
Author(s) -
Andersson Maria,
Janosik Tomasz,
Shirani Hamid,
Slätt Johnny,
Fischer Andreas,
Beck Olof
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100739
Subject(s) - chemistry , chromatography , morphine , sulfate , urine , bioanalysis , coefficient of variation , pharmacology , biochemistry , medicine , organic chemistry
The aim of this work was to synthesize morphine‐3‐ O ‐sulfate and morphine‐6‐ O ‐sulfate for use as reference substances, and to determine the sulfate conjugates as possible heroin and morphine metabolites in plasma and urine by a validated LC‐MS/MS method. Morphine‐6‐ O ‐sulfate and morphine‐3‐ O ‐sulfate were prepared as dihydrates from morphine hydrochloride, in overall yields of 41 and 39% with product purities of >99.5% and >98%, respectively. For bioanalysis, the chromatographic system consisted of a reversed‐phase column and gradient elution. The tandem mass spectrometer was operated in the positive electrospray mode using selected reaction monitoring, of transition m / z 366.15 to 286.40. The measuring range was 5–500 ng/mL for morphine‐3‐ O ‐sulfate and 4.5–454 ng/mL for morphine‐6‐ O ‐sulfate in plasma. In urine, the measuring range was 50–5000 ng/mL for morphine‐3‐ O ‐sulfate and 45.4–4544 ng/mL for morphine‐6‐ O ‐sulfate. The intra‐assay and total imprecision (coefficient of variation) was below 11% for both analytes in urine and plasma. Quantifiable levels of morphine‐3‐ O ‐sulfate in authentic urine and plasma samples were found. Only one authentic urine sample contained a detectable level of morphine‐6‐ O ‐sulfate, while no detectable morphine‐6‐ O ‐sulfate was found in plasma samples.