Monitoring of morphine in urine after consumption of opium poppy sprouts using immunoassay and liquid chromatography coupled with quadrupole exactive Orbitrap mass spectrometry

• Opiates were detected in urine following the consumption of fresh opium poppy sprouts. • Poppy sprouts produced detectable morphine leves in the urine of most participants. • Morphine remained detectable in urine for up to 24 hours after poppy sprout ingestion. • The findings emphasise the need for careful interpretation of opiate testing in forensic settings. Opiate use remains a growing global concern, particularly in the context of drug testing and forensic toxicology, where the differentiation between illicit drug use and dietary exposure continues to pose a significant interpretative challenge. Poppy seed–based products on opiate drug test results have been extensively investigated, although the impact of consuming fresh opium poppy sprouts, locally known as “afiyan,” hasn’t previously been evaluated. This study aimed to investigate whether the consumption of fresh poppy sprouts could result in presumptive positive opiate results and to assess the implications for forensic toxicological interpretation.Ten healthy volunteers who regularly consumed this herb were included in the study. Urine samples were collected over a 48h period under non-fasting conditions. Opiate screening was performed using immunoassay at two cut-off concentrations (300 and 2000ng/mL). Quantitative analysis of morphine, codeine, thebaine, norcodeine, and dihydrocodeine was conducted using a validated Orbitrap LC–MS/MS method (linearity:30–1200 ng/mL; LLOQ: 30ng/mL for all analytes). At the 300ng/mL screening cut-off, 70% (n=7) of participants yielded positive results within the first hours following ingestion. Morphine concentrations exceeded the LLOQ in first-hour urine samples from nine participants and persisted for up to 12 h in eight participants (37.95–150.23ng/mL). Morphine was still detected after 24 h in five participants (44.38–113.4ng/mL). These findings demonstrate that consumption of fresh poppy sprouts can lead to detectable urinary morphine concentrations capable of triggering positive opiate screening results. This work provides novel analytical evidence on the alkaloid profile and forensic relevance of fresh poppy sprout consumption.