Development of a Method to Extract Opium Poppy (Papaver somniferum L.) DNA from Heroin

Abstract

This study is the first to report the successful development of a method to extract opium poppy (Papaver somniferum L.) DNA from heroin samples. Determining of the source of an unknown heroin sample (forensic geosourcing) is vital to informing domestic and foreign policy related to counter-narcoterrorism. Current profiling methods focus on identifying process-related chemical impurities found in heroin samples. Changes to the geographically distinct processing methods may lead to difficulties in classifying and attributing heroin samples to a region/country. This study focuses on methods to optimize the DNA extraction and amplification of samples with low levels of degraded DNA and inhibiting compounds such as heroin. We compared modified commercial-off-the-shelf extraction methods such as the Qiagen Plant, Stool and the Promega Maxwell-16 RNA-LEV tissue kits for the ability to extract opium poppy DNA from latex, raw and cooked opium, white and brown powder heroin and black tar heroin. Opium poppy DNA was successfully detected in all poppy-derived samples, including heroin. The modified Qiagen stool method with post-extraction purification and a two-stage, dual DNA polymerase amplification procedure resulted in the highest DNA yield and minimized inhibition. This paper describes the initial phase in establishing a DNA-based signature method to characterize heroin.

New EST-SSR Markers for Individual Genotyping of Opium Poppy Cultivars (Papaver somniferum L.)

“…Thus, P. somniferum (2n = 22) could be classified as an alloaneuploid, where its basic chromosome number x = 11 is derived from the more ancestral x = 7, given the existence of a hypothetical triploid hybrid that may have preceded the speciation of P. somniferum [9]. Although Marciano et al [6] mentioned that the diploid nature of P. somniferum simplified the workflow for genetic identification and compared it with the development of markers for human identification, we worried that they probably underestimated the evolutionary complexity of the genome development typical for flowering plants and the consequences of allopolyploidy [44,45]. Namely, for P. somniferum, analysis of a 401 kb long genomic sequence of gene cluster for noscapine synthesis was performed, revealing gene duplicities, structural rearrangement, and many DNA or retrotransposable elements [5].…”

“…Firstly, P. somniferum serves as a source of secondary metabolites such as morphine, thebaine, codeine, or noscapine, which are utilized by pharmaceutical companies for producing medicines with analgesic, antitussive, sedative, or anti-tumor effects [4,5]. Its dark side is infamously the abuse of its sap ("opium latex") for the production of heroin [6]. Secondly, poppy seeds do not contain a high amount of morphinan alkaloids when they are properly harvested and/or treated [7,8]; thus they are also used in the food industry, typically in Central and Eastern Europe, given their historical and cultural background.…”

“…Nevertheless, it seems that these methods are not sufficiently reliable at the fine scale necessary for the discrimination of individual varieties. This is probably due to the complex influence of many environmental conditions changing the alkaloid content [6,17,18] leading to a great intra-cultivar variability. Here, DNA markers, which are not influenced by these factors, have proven to be helpful.…”

Scientific Prospects for Cannabis-Microbiome Research to Ensure Quality and Safety of Products

“…In this regard, systems biology [1,2] is well-equipped to integrate findings and define the genetic variability and functional metabolites [3] in a model plant system. Model plant research, such as that of well-known Arabidopsis thaliana [4][5][6][7] and the extensively-studied medicinal plant, Papaver somniferum [8][9][10][11][12], highlight the significance of gathering information about regulation of plant fitness and homeostatic mechanisms of metabolites [13] orchestrated by the host genome and its affiliated microbial metagenome [14]. Such an approach notably reveals the importance of the plant's genome-microbiome interactions [15], which changes under stress or disease conditions [16,17] as plants selectively source their microbiomes to suit their needs [18,19].…”

Selection of the Reference Gene for Expression Normalization in Papaver somniferum L. under Abiotic Stress and Hormone Treatment

“…Due to their analgesic activities, pharmaceutical opiates are universally applied for relieving the pain caused by cancer, surgery, and wounds [3]. The opium poppy is also a source of narcotics, such as opium and heroin [4]. Consequently, many countries forbid or strictly control the cultivation of P. somniferum and the uses of opium products through legislation.…”