Gas Chromatographic/Mass Spectrometric Analysis of Morphine and Codeine in Human Urine of Poppy Seed Eaters

Abstract

In this study, poppy seeds were examined for a natural constituent that might serve as a marker for the seeds' ingestion as opposed to opiate abuse. Thebaine was selected as possible marker, since it was found to be a component of all poppy seeds examined and was not a natural component of different heroin samples. During the course of this investigation, a new extraction and cleanup procedure was developed for the gas chromatographic/nitrogen phosphorus detection (GC/NPD) and gas chromatographic/mass spectrometric (GC/MS) analysis of morphine and codeine in urine. A linear response, over a concentration range of 25 to 600 ng/mL, was obtained for codeine and morphine (r = 0.9982 and 0.9947, respectively). The minimum detectable level (LOD) and limit of quantitation (LOQ) for morphine were 10 and 30 ng/mL, respectively; whereas LOD and LOQ for codeine were 2 and 8 ng/mL, respectively. The coefficients of variance (CV, n = 6) for morphine and codeine analyses at the 100-ng/mL level were 13.3 and 4.6%, respectively. This procedure was used for the analysis of urine samples from five poppy seed eaters who each ingested 200 g of poppy seed cake. Results indicated that significant amounts of morphine and codeine are excreted in urine and that in all subjects, at least at one point in time, the apparent morphine concentration as determined by radioimmunoassay (RIA) analysis exceeded the cutoff value (300 ng/mL) established for screening. Thebaine was not detected in urine specimens collected following poppy seeds ingestion and thus could not be used as a marker.

GC-MS detection and characterization of thebaine as a urinary marker of opium use

“…Both mass spectra were characterized as thebaine by database mass spectral libraries due to its small size, the TMS group (electrophile) has probably reacted with the hydrogen at position 5 of thebaine in the present work. It should be noted that the formation of TMS derivative of thebaine could not be realized earlier by some workers [12] who reported the absence of this marker compound in the urine samples of poppy seed users. These authors employed acetyl derivatization for determining morphine and codeine in urine using acetic anhydride, a reagent known to cause degradation of thebaine to acetyl thebaol and/or 3,6-dimethoxy-4,5-epoxy-phenanthrene [25].…”

“…Cassella et al [13] and Wu and Dobberstein [29] also reported a heat effect on decomposition of the alkaloid. On the other hand, thebaine was found to be stable in simulated gastric medium [12]. Therefore, it can be stated with a fair amount of certainty that the marker compound thebaine will be detected in most urine samples when opium is ingested by the oral route.…”

“…Cassella et al [13] reported positive results, while ElSohly et al [12] reported negative ones. Also at variance were reports on the amenability of thebaine to trimethylsilyl (TMS) derivatization.…”

“…1), a constituent of both opium and poppy seeds, had been suggested as a urinary marker to differentiate between heroin use and the use of poppy seed-containing foods [12][13][14][15][16], which are common in North America and western European countries. Because thebaine is present at much higher concentrations in opium (0.1-2.5% w/w) [17] than in the poppy seeds (0.00003-0.0014% w/w) [12], it should be targeted as a marker of opium use. This targeting constitutes one of the main subjects of this study.…”

Summer Research Program - 1996. Summer Faculty Research Program. Final Reports. Volume 2B, Armstrong Laboratory

“…(c) The effects of acute ingestion of poppy seed pastries on the urinary excretion of morphine and codeine has been extensively reported ( Fritschi and Prescott Jr,1985;Hayes et al,1987;Pettitt et 41-4 al.,1987;Struempler,1987;ElSohly et al, 1988;ElSohly and Jones,1989;ElSohly and ElSohly, 1990;Selavka, 1991;Carpenter, 1994;Huestis and Cone, 1995). This data provides information on the maximum excretion of these two alkaloids following a one time ingestion.…”