The poppy seed defense: a novel solution

Abstract

A major toxicological challenge is distinguishing whether morphine in urine, in the absence of 6-monoacetylmorphine (6-MAM), originates from 'street' heroin use or poppy seed ingestion. Manufacturing byproducts from the synthesis of illicit heroin include those that originate from the reaction of acetic anhydride with the alkaloid impurity, thebaine, which undergoes skeletal rearrangement, resulting in compounds with a 2-(N-methylacetamido)ethyl side-chain. The hypothesis that the tertiary amide in this side-chain is resistant to endogenous hydrolysis was supported from in-vitro experiments; a glucuronide metabolite (designated 'ATM4G') was identified that may be used as a marker of 'street' heroin administration. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis for this metabolite was then performed on selected urine specimens from 22 known heroin users, these being negative on routine testing for 6-MAM by gas chromatography-mass spectrometry (GC-MS), using the generally applied reporting threshold of 10 ng/mL, but positive for the presence of morphine. Peaks corresponding to the retention time for the metabolite marker were clearly observed for 16 of the 22 samples, with variations of the ratios of its three dependent ions being within ± 30% of that produced in vitro. Conversely, 6-MAM was detected in only 3 samples, but at concentrations <1 ng/mL. Such a high frequency for the presence of the metabolite marker in urine, in the absence of 6-MAM, is noteworthy and suggests that detection of this metabolite may offer an important advance in forensic toxicology, allowing the development of a new and more definitive test for heroin abuse and thus a potential solution to the so-called 'poppy seed defense'.

Morphine and codeine concentrations in human urine following controlled poppy seeds administration of known opiate content

“…Others reported markers in heroin, such as acetylcodeine, with a similar outcome [3,14]. A recent report by Chen et al appears more promising [15]. These investigators employed a sensitive liquid chromatography tandem mass spectrometric method to identify an acetylated thebaine glucuronide (3,6-dimethoxy-4-acetoxy-8-[2-(N-methylacetamido)ethyl]phenanthrene; abbreviated ATM4G) in 16 of 22 urine specimens produced by 22 heroin addicts who had 6-AM <10μg/L in each specimen and total morphine (morphine-3-glucuronide and morphine) concentrations >300μg/L in 17 of the specimens.…”

Urinary excretion study following consumption of various poppy seed products and investigation of the new potential street heroin marker ATM4G

“…Moreover, poppy seed ingestion study performed by Chen et al [8] revealed similar results even following consumption of poppy seeds containing up to 60 μg thebaine. Consequently, the prerequisites for the formation of ATM4G following consumption of poppy seed products were met.…”

“…[8] Poppy seed products and alkaloid extraction Unground blue poppy seeds were obtained from Rapunzel Naturkost GmbH (Legau, Germany), Edeka (Hamburg, Germany) and Seeberger (Ulm, Germany). The reference materials 6-AC, papaverine, noscapine, 6-MAM, morphine, and codeine as well as the internal standards 6-MAMd 3 , morphine-d 3 , and codeine-d 3 were obtained from Cerilliant (Round Rock, TX, USA).…”

“…Following administration, this precursor is deacetylated in phase I and then conjugated with glucuronic acid in phase II metabolism, resulting in the formation of ATM4G (Figure 1). [8] The aim of this recent study is to confirm the assumption that ATM4G is not metabolically formed and excreted via urine after consumption of different poppy seed products sold in Germany. Furthermore, it can be expected, that based on the complex formation process of this compound, it will not be metabolically formed after the ingestion of thebaine-containing poppy seed products.…”

Morphine and codeine in oral fluid after controlled poppy seed administration

“…Despite multiple efforts, no unambiguous markers were identified, nor has a poppy seed consumption limit or analytical cutoff been developed to differentiate poppy seed ingestion from heroin or morphine use. A recent study suggested that urinary ATM4G, a glucuronide metabolite of a byproduct from the synthesis of illicit heroin (thebaine with a 2(N-methylacetamido)ethyl side-chain), can differentiate the source of opiate intake [5]. …”