Evaluation of the mass spectrometric fragmentation of codeine and morphine after 13C-isotope biosynthetic labeling

How human neuroblastoma cells make morphine

“…However, in mammals, norlaudanosoline has been detected on few occasions: in rat striatum (15), human brain (16), a human pancreatic cell line (1), and humans treated with various pharmacological agents, for instance alcohol and L-dopa (17). In agreement with these findings, we have proven the endogenous origin of norlaudanosoline in human cell cultures by our previous experiments showing position specific labeling with 18 O and norlaudanosoline being part of the (S)-reticuline pathway in DAN-G cells (1).…”

“…In a similar application experiment, the neuroblastoma cells were cultured in the presence of 20 M [2,2-2 H 2 ]dopamine, resulting in a 22% isotopic enrichment of this precursor in morphine. According to the characteristic MS fragmentation pattern of morphine, as investigated in our laboratory (18,19), the dopamine molecule resided exclusively in the cyclohexene ring of morphine together with C-15, C-16, and the nitrogen atom ( Table 1 (Fig. 2).…”

“…In previous studies, by using the least invasive isotopic precursor of morphine, 18 O, nonrandom, position-specific incorporation of two atoms of 18 O into one molecule of morphine produced by the living, metabolizing neuroblastoma cells (at a concentration of 10 nM) could be demonstrated in our laboratory. Moreover, the potential precursor alkaloids norlaudanosoline (tetrahydropapaveroline) and (S)-reticuline, which were also produced by these cells, were shown to be labeled with two atoms of 18 O (1).…”

“…This knowledge of the morphine biosynthesis in plants and the methodology of its elucidation were now applied to investigate the origin and the biosynthetic pathway of morphine found in mammalian cells. In brief, heavy isotope-labeled precursors (e.g., with 13 C, 2 H, and 18 O labels) are administered to human cell cultures and incubated for a certain period. Subsequently, the desired products are isolated, and their structure, isotope content, and labeling pattern, which has to be nonrandom, are elucidated by spectroscopic methods.…”

Trace-Level Screening of Chemicals Related to Clandestine Desomorphine Production with Ambient Sampling, Portable Mass Spectrometry

“…Representative DESI-MS and PSI-MS data for the desomorphine precursor, codeine, can be seen in Figure 2. Both ionization sources yield a single parent ion, [codeine + H] + at / 300 (Figures 2(a) and 2(c)), with characteristic fragments at / 183, 215, 225, and 243 corresponding to proposed losses of C 13 H 11 O, C 4 H 7 NO, C 3 H 9 NO, and C 3 H 7 N, respectively (as seen in Figures 2(b) and 2(d)); these MS 2 transitions and associated losses are similar to those reported in the literature [26]. Of note, comparison of the MS spectral intensities obtained for the target analytes using both ionization methods demonstrates higher signal intensity with PSI for both analytes.…”