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. 2014 Dec;2(6):e00071.
doi: 10.1002/prp2.71. Epub 2014 Sep 1.

Detection of morphine-3-sulfate and morphine-6-sulfate in human urine and plasma, and formation in liver cytosol

Maria Andersson  1 , Linda Björkhem-Bergman  2 , Lena Ekström  1 , Lena Bergqvist  3 , Hugo Lagercrantz  3 , Anders Rane  1 , Olof Beck  1
Affiliations

Detection of morphine-3-sulfate and morphine-6-sulfate in human urine and plasma, and formation in liver cytosol

Maria Andersson et al. Pharmacol Res Perspect. 2014 Dec.

Abstract

Morphine is still the mainstay in treatment of severe pain and is metabolized in the liver mainly by glucuronidation, partly to the pharmacologically active morphine-6-glucuronide (M6G). The sulfation pathway has attracted much less attention but may also form active metabolites. The aim of the present study was to study two sulfate metabolites of morphine in humans. Urine and plasma from newborns, adult heroin addicts, and terminal cancer patients was analyzed for the presence of morphine-3-sulfate (M3S) and morphine-6-sulfate (M6S) by a new liquid chromatography - tandem mass spectrometry (LC-MS/MS) method. In addition, morphine sulfation was studied in vitro in human liver cytosol preparations. M3S was present in urine and plasma from all study groups although at lower concentrations than morphine-3-glucuronide (M3G). The plasma M3S/M3G ratio was 30 times higher in newborns than in adults indicating that the relative sulfation is more important at early stage of life. M6S was measurable in only one plasma sample from a newborn patient, and in one of the urine sample from the drug testing group. The incubation of morphine with liver cytosol extracts resulted in approximately equal rate of formation of both M3S and M6S. In conclusion, sulfation of morphine is catalyzed in human liver but this minor metabolic pathway probably lacks clinical significance. The M6S metabolite is formed at a low rate, making it undetectable in most individuals.

Keywords: LC-MS/MS; morphine; morphine-3-sulfate; morphine-6-sulfate; plasma; urine.

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Figures

Figure 1
Figure 1
Calibration curves were generated between 14 and 14,000 nmol/L for the morphine sulfates. An LC-MS/MS chromatogram is displayed of a plasma calibrator spiked with 275 nmol/L of M3S and M6S, respectively.
Figure 2
Figure 2
Analysis of M3S and M6S in authentic urine and plasma samples using LC-MS/MS. Chromatograms showed are (A) a plasma sample from the newborn group containing 20 nmol/L M6S and a detectable peak for M3S and (B) urine sample from the drug testing group containing 160 nmol/L M6S and 4000 nmol/L M3S.
Figure 3
Figure 3
A second order polynomial graph displaying the M3S/M3G ratio for the newborn group compared to their gestational age at birth. A tendency can be observed that preterm newborns have higher ratios than full-term newborns.
Figure 4
Figure 4
Formation of M3S and M6S in human liver cytosol was studied in vitro. An LC-MS/MS chromatogram is displayed of a morphine incubation with adult liver cytosol containing 20 nmol/L M6S and 8 nmol/L M3S.
See this image and copyright information in PMC

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