Drug Testing and Analysis最新文献

{"title":"Ethyl Glucuronide in Hair: A 10-Year Overview","authors":"Sara Casati,&nbsp;Alessandro Ravelli,&nbsp;Roberta F. Bergamaschi,&nbsp;Caterina Fabbri,&nbsp;Erika Palmisano,&nbsp;Andrea Ostinelli,&nbsp;Gabriella Roda,&nbsp;Giuseppe Facchi,&nbsp;Marica Orioli","doi":"10.1002/dta.70041","DOIUrl":"10.1002/dta.70041","url":null,"abstract":"<p>This 10-year retrospective study evaluates hair ethyl glucuronide (EtG), a direct metabolite of ethanol, in a large Northern Italian cohort (<i>N</i> = 68,221 samples collected between 2013 and 2022). The analysis aimed to evaluate long-term alcohol consumption through the EtG distribution in terms of age, gender, recidivism, applicants, sampling region, sampling length, and the impact of seasonality and the COVID-19 pandemic. Hair EtG was determined by HPLC-MS/MS, and samples were classified according to the Society of Hair Testing (SoHT) recommended cut-offs: EtG ≤ 5 pg/mg (does not contradict abstinence), 5 &lt; EtG &lt; 30 pg/mg (repeated alcohol consumption), and EtG ≥ 30 pg/mg (chronic excessive alcohol consumption). A descriptive sensitivity analysis using SoHT cut-offs was performed to allow international comparability. The percentage (%) of hair samples classified as repeated or chronic excessive alcohol consumption was 13.6% (<i>N</i> = 9,281) and 5.6% (<i>N</i> = 3,818), respectively. EtG values significantly varied with gender and age as well as referral context. We observed statistically significant differences in EtG concentrations recorded in head, chest, axillary, and pubic hair samples. Significantly higher EtG values were detected in 3-cm proximal head hair versus the 3- to 6-cm proximal segment as well as across seasons, with a higher concentration in colder months. Conversely, no measurable short-term population impact of COVID-19 was revealed by hair EtG, whereas a significant long-term influence was highlighted. Overall, despite a large variability of EtG concentrations, this study provides robust evidence of the reliability of hair EtG testing and offers a comprehensive overview of long-term alcohol consumption trends in a monitored Italian population.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"518-529"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040436/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146218019","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Isomer Composition Assessment of Synthetic Phosphatidylethanol by Collision-Induced and Ozone-Induced Dissociation Mass Spectrometry","authors":"Matthias Bantle,&nbsp;Jackson O. T. Long,&nbsp;Samuel C. Brydon,&nbsp;Reuben S. E. Young,&nbsp;Stephen J. Blanksby,&nbsp;Wolfgang Weinmann,&nbsp;Marc Luginbühl","doi":"10.1002/dta.70046","DOIUrl":"10.1002/dta.70046","url":null,"abstract":"<p>The direct blood-alcohol biomarker phosphatidylethanol (PEth), especially its most abundant analogue 1-palmitoyl-2-oleoyl-<i>sn</i>-phosphatidylethanol (PEth 16:0/18:1) has gained increasing relevance in clinical and forensic applications for assessing alcohol consumption. Accurate quantification of PEth is essential to reliably differentiate between abstinence, moderate alcohol consumption and excessive alcohol intake. Measurement accuracy of PEth 16:0/18:1 by well-established liquid chromatography–tandem mass spectrometry (LC-MS/MS) approaches such as multiple reaction monitoring (MRM) can be confounded by the presence of the regioisomer 1-oleoyl-2-palmitoyl-<i>sn</i>-phosphatidylethanol (PEth 18:1/16:0) in samples and synthetic reference standards. To address this measurement uncertainty, we conducted a new assessment of the isomeric composition of six currently available reference materials from four suppliers using collision-induced dissociation/ozone-induced dissociation (CID/OzID). Examination of these synthetic compounds found a high degree of regioisomeric purity of &gt; 95%. Thus verified, the relative abundance of two key LC–MS/MS transitions were compared across a range of collision energies for both reference materials and an exemplary set of 10 dried blood spot case samples. These findings suggest a significantly wider range of natural isomer distributions spanning both higher and lower regiochemical composition (88.8%–98.85%) than the reference materials but within a range that would not significantly impact clinical classification.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"501-509"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040422/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146211565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Concentration and Detection Time of Nitrous Oxide in Blood Following Controlled Inhalation","authors":"Angelica Ørregaard Lindholm,&nbsp;Marie Katrine Klose Nielsen,&nbsp;Mette Kristensen,&nbsp;Mo Haslund Larsen,&nbsp;Johan Heiberg,&nbsp;Brian Schou Rasmussen","doi":"10.1002/dta.70053","DOIUrl":"10.1002/dta.70053","url":null,"abstract":"<p>Recreational use of nitrous oxide (laughing gas, N₂O) is becoming increasingly common and abuse is often seen in relation to driving, posing significant traffic-safety concerns. Only a few studies exist on blood concentrations of N₂O and its detectability over time after use. In this study, 11 volunteers received controlled administration of 50% N₂O for 10 min, after which blood samples were drawn and analyzed for N₂O by headspace-gas chromatography–mass spectrometry (HS-GC–MS). Pharmacokinetic modelling indicated that elimination of N₂O can best be described by a two-compartment model with half-lives of 2.4 and 31 min. Although both the pharmacological effect and intoxication typically disappear within minutes after intake, N₂O remained detectable in blood for an average of 62 min at a cutoff of 0.2 mL/L and 132 min at a cutoff of 0.05 mL/L.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"576-580"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040437/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147323938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Evaluating Transcriptomic Biomarkers for rHuEPO Detection: Assessing the Impact of Exercise and Altitude Exposure","authors":"Daria Obratov,&nbsp;Shaun Sutehall,&nbsp;Longhua Liu,&nbsp;Zhao Zhongying,&nbsp;Yannis Pitsiladis","doi":"10.1002/dta.70040","DOIUrl":"10.1002/dta.70040","url":null,"abstract":"<p>Recombinant human erythropoietin (rHuEPO) is often misused in endurance sports due to its potent erythropoietic effects. While transcriptomic biomarkers hold promise for detecting rHuEPO use beyond conventional testing windows, many proposed gene markers may also respond to physiological stimuli such as exercise or altitude. This study compared 153 previously reported rHuEPO-responsive genes in whole blood with transcripts identified during exercise (GEPREP database) and high-altitude exposure (four independent studies). For the exercise dataset, gene-level statistical outputs were obtained directly from the GEPREP database, while biological relevance was calculated using Cohen's <i>d</i>. Analyses of altitude and rHuEPO datasets followed the original statistical procedures described in each study. Among the 153 rHuEPO-responsive genes, 94 overlapped with altitude and 34 with exercise. However, 50 genes remained unaffected by either exercise or altitude stimuli. Enriched in post-translational regulation and intracellular transport pathways, these genes represent promising candidate transcriptomic markers of rHuEPO administration. This work provides a refined gene panel that reduces the likelihood of false positives and requires further experimental validation before integration into RNA-based detection tests.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"581-602"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040416/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147363395","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In Vitro Metabolism of 1-Benzoyl-Lysergic Acid Diethylamide (1Bz-LSD) and Identification of a Deethylated Metabolite (1Bz-LAE) Using a Synthesized Reference Standard","authors":"Yuki Azuma,&nbsp;Akiko Asada,&nbsp;Misa Tanaka,&nbsp;Takahiro Doi","doi":"10.1002/dta.70042","DOIUrl":"10.1002/dta.70042","url":null,"abstract":"<div>\u0000 \u0000 <p>Lysergic acid diethylamide (LSD) analogs represent an emerging class of new psychoactive substances (NPS). These compounds are often rapidly metabolized, making direct detection of the parent compound in biological samples difficult. Therefore, metabolite detection serves as a critical and effective strategy for the confirmation of consumption. However, the metabolic pathways of these analogs remain largely uncharacterized, and synthesized metabolite standards for definitive identification are mostly unavailable. This study investigated the newly emerged LSD analog, (8<i>R</i>)-1-benzoyl-<i>N,N</i>-diethyl-6-methyl-9,10-didehydroergoline-8-carboxamide (1-benzoyl-LSD, 1Bz-LSD), which features a benzoyl moiety at the <i>N</i>¹ position. The objectives were to characterize its metabolic fate and synthesize a targeted metabolite analyte for the confirmation of consumption. In vitro metabolism of 1Bz-LSD was examined using human liver microsomes and analyzed through liquid chromatography–quadrupole time-of-flight mass spectrometry. The parent compound was rapidly metabolized, yielding 15 metabolites, including LSD. Among these, a deethylated metabolite was detectable over a prolonged period, indicating its potential as a targeted analyte. To confirm its structure, this metabolite was chemically synthesized and identified as 1-benzoyl-lysergic acid ethylamide (1Bz-LAE). The synthesis and identification of 1Bz-LAE provide a crucial analytical foundation for the investigations of 1Bz-LSD use, with this metabolite anticipated to function as a valuable marker.</p>\u0000 </div>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"490-500"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146206134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Annual Banned-Substance Review 18th Edition—Analytical Approaches in Human Sports Drug Testing 2024/2025","authors":"Mario Thevis,&nbsp;Tiia Kuuranne,&nbsp;Hans Geyer","doi":"10.1002/dta.70033","DOIUrl":"10.1002/dta.70033","url":null,"abstract":"<p>Alongside the considerable advances and accomplishments in drug research and development, the breadth of anti-doping research topics has also continued to grow. This is particularly relevant not only to provide comprehensive information on new drug entities, drug metabolism and elimination, and/or the impact of administration and exposure routes on analytical test results but also to ensure the timely implementation of analytical methods that ensure the availability of relevant anti-doping testing procedures and corresponding analytical data for routine doping control applications.</p><p>The 18th edition of the annual banned-substance review on analytical approaches in human sports drug testing is dedicated to literature published between October 2024 and September 2025, and information published within these 12 months on established doping agents as well as new (potentially and evidently) relevant substances is reviewed and discussed, especially in the context of the World Anti-Doping Agency's 2025 Prohibited List.</p><p>Topics of particular interest have been investigations into the metabolic fate and detection of anabolic agents, both anabolic–androgenic steroids and other anabolic substances such as selective androgen receptor modulators and protein-based therapeutics negatively regulating the activin receptor signaling pathway, and detection strategies for numerous new drug candidates have been discussed and presented. Further, the trend toward expanding testing options with regard to gene doping practices including oligonucleotide-based compounds (e.g., small interfering RNA, antisense oligonucleotides, etc.), transgenes, and gene editing practices continued also in 2024/2025, underlining the relevance for current and future sports drug testing programs.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"458-482"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040425/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146211522","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Detection of a Distinct Erythropoietin (EPO) Profile After Isoelectric Focusing in Patients With Familial Erythrocytosis","authors":"Laurent Martin,&nbsp;Salam Idriss,&nbsp;Nada Maaziz,&nbsp;Soizic Huerre,&nbsp;Lamine Redouane Mekacher,&nbsp;Kahina Lahmek,&nbsp;François Girodon,&nbsp;Betty Gardie,&nbsp;Magnus Ericsson,&nbsp;Alexandre Marchand","doi":"10.1002/dta.70049","DOIUrl":"10.1002/dta.70049","url":null,"abstract":"<p>Since the implementation of the first erythropoietin (EPO) detection method to fight the use of recombinant EPO (rEPO) for doping, the techniques used and identification criteria have evolved. The first technique involved iso-electric focusing (IEF-PAGE) to separate EPO isoforms by their charges. It can still be used for initial testing but is now largely replaced by electrophoretic separation based on molecular weight (SDS-PAGE or SAR-PAGE) to conclude decisively on the presence of rEPO. Recently, different mutations in the noncoding region of the <i>EPO</i> gene were identified in patients with familial erythrocytosis. Their blood EPO profiles after IEF-PAGE were very basic and in the same area as rEPO. To avoid any possible misinterpretation in an antidoping context, the goal of this work was to characterize urine and blood EPO profiles from these patients using all methods authorized in antidoping laboratories (IEF-PAGE, SDS-PAGE, and SAR-PAGE) and to compare them with wild-type (WT) EPO and with profiles obtained after the administration of rEPO Eprex. Results showed that SDS-PAGE and SAR-PAGE profiles were not modified in the patients compared with WT EPO and could not be mistaken for the presence of rEPO. In contrast, the IEF-PAGE profiles in urine were slightly more acidic than in blood and closer to the rEPO profile. However, they did not meet the identification criteria used when this technique was the only authorized method. It is highly unlikely that these <i>EPO</i> mutations, only identified in erythrocytosis patients yet, could lead to an adverse analytical finding.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"537-542"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040431/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146224860","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analytical Challenges in Detecting New Psychoactive Substances in Anti-Doping: A Proof-of-Concept Involving Pentedrone Metabolite","authors":"D. Kwiatkowska,&nbsp;B. Peta,&nbsp;K. Grucza,&nbsp;A. Łabęcka,&nbsp;P. Konarski,&nbsp;K. Wojtkowiak,&nbsp;K. Grela,&nbsp;A. Sytniczuk,&nbsp;M. Wicka","doi":"10.1002/dta.70047","DOIUrl":"10.1002/dta.70047","url":null,"abstract":"<p>New psychoactive substances (NPS) have become increasingly prevalent worldwide, posing a growing challenge to public health, forensic science, and doping control. Their structural diversity, rapid emergence, and legal ambiguity complicate both toxicological assessment and analytical detection. Research into the metabolism of these compounds is limited by ethical constraints on administration studies, hindering the development of reference standards. This study presents a methodological approach for the identification and confirmation of 2-amino-1-phenylpentan-1-ol, a metabolite associated with several synthetic cathinones, including pentedrone, <i>α</i>-pyrrolidinopentiophenone (PVP), and <i>N</i>-ethylpentedrone (NEP). The compound was detected in a routine doping control sample using ultra-performance liquid chromatography–tandem mass spectrometry (UPLC–MS/MS). In the absence of a commercially available certified reference material (CRM) encompassing all potential isomeric forms, a custom synthesis of the target metabolite was undertaken. The synthesized CRM-enabled unambiguous confirmation of the analyte and its diastereomeric composition, demonstrating the importance of reference compound availability for the accurate interpretation of results in anti-doping analysis. This work highlights both the analytical challenges posed by NPS and the need for continued development of tailored CRMs to support doping control laboratories worldwide.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":"18 4","pages":"530-536"},"PeriodicalIF":2.7,"publicationDate":"2026-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13040411/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146224854","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"In Vivo and In Vitro Evaluation of PTeCA (1H-Pyrrole-2,3,4,5-Tetracarboxylic Acid) in Hair Matrix as a Marker for Oxidative Cosmetic Treatment.","authors":"Sara Casati, Alessandro Ravelli, Roberta F Bergamaschi, Agata Magliocco, Sofia Vanerio, Gabriella Roda, Paola Rota, Marica Orioli, Alessio Battistini","doi":"10.1002/dta.70068","DOIUrl":"https://doi.org/10.1002/dta.70068","url":null,"abstract":"<p><p>Alteration of the hair matrix by cosmetic products presents a challenge for forensic hair analysis. Oxidative treatments lead to analyte depletion and false-negative results. Currently, the degradation product of eumelanin, 1H-pyrrole-2,3,5-tricarboxylic acid (PTCA) is being investigated as a marker for oxidative hair treatment; however, it requires the definition of the cut-off value. Recently, it has been shown that 1H-pyrrole-2,3,4,5-tetracarboxylic acid (PTeCA) also increased significantly after in vitro oxidative hair treatments. Here, our previously published LC-MS/MS method for hair PTCA has been fully validated for the simultaneous quantification of PTeCA (range 0.01-2.5 ng/mg; lower limit of quantification [LLOQ] 0.003 ng/mg). The method was applied to 3378 self-reported treated (T) and untreated (UT) hair samples (3-6 cm proximal). In addition, the in vitro formation of PTeCA was assessed in 225 UT hair samples by different professional cosmetic treatments with and without oxidative agents. In the UT group (N = 1144), PTCA was determined in about 40% of the samples with a median PTCA of 0.04 ng/mg (range 0.01-14.9 ng/mg) and PTeCA was >LLOQ in < 2% of the samples (N = 53). In the T group (N = 425), PTCA was determined in 84% of the samples with a median of 0.75 ng/mg (range 0.01-58.1 ng/mg); while the median PTeCA was 0.40 ng/mg (N = 243; range 0.02-31.2 ng/mg). Moreover, the in vitro cosmetic treatment confirmed the PTeCA formation only in oxidative conditions. Finally, a PTCA cut-off value was proposed using PTeCA as the gold standard. Our data suggest that PTeCA could be a reliable marker for detecting oxidative cosmetic treatments in the hair matrix.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147588938","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effect of Reaction Time and Temperature on the Impurity Profile of Amphetamine in the Leuckart Method.","authors":"Maartje C M Verhoeven, Robert C Huls, Nelly W van der Stelt, Jelle C de Koning, Marco van Grol, Ruben F Kranenburg, Arian C van Asten, Janneke W Hulshof","doi":"10.1002/dta.70063","DOIUrl":"https://doi.org/10.1002/dta.70063","url":null,"abstract":"<p><p>Illicit amphetamine synthesis is frequently conducted using the well-known Leuckart method, generating complex impurity profiles that provide forensic insights. This study reports the effect of reaction temperature and time during the first step of the Leuckart synthesis on subsequent conversion efficiency and impurity profile. Controlled syntheses were conducted at 120°C, 140°C, and 160°C, with samples collected at defined intervals. Reaction progress was monitored by gas chromatography-flame ionization detection (GC-FID), and impurity profiling followed the European harmonized method for profiling amphetamine (EHMPA) using GC-mass spectrometry (GC-MS). In addition, preparative liquid chromatography (prep-LC) followed by LC-high-resolution MS (LC-HRMS) was employed to isolate and characterize previously unidentified impurities. Results indicate that higher temperatures and longer reaction times accelerate benzyl methyl ketone (BMK) conversion, increasing the formation of N-formylamphetamine (after step 1) and amphetamine (after step 2). Lower temperatures and shorter reaction times result in incomplete conversion and the persistent presence of BMK, along with several impurities, including unsaturated ketones (3,5-diphenyl-4-methyl-3-penten-2-one and 1,5-diphenyl-4-methyl-3-penten-2-one), 1,3-dimethyl-2-phenylnaphthalene (Naphthalene 1), and 1-benzyl-3-methylnaphthalene (Naphthalene 2). Elevated temperatures favor the formation of 4-methyl-5-phenylpyrimidine (4M5PP), 4-benzylpyrimidine (4BP), N,N-bis-(‒1-phenylpropan-2-yl)formamide (DPIF), and Unknowns A2, B1, and B2, which were tentatively identified as N-[3,5-diphenyl-4-methyl-3-penten-2-yl]amine, and the cis/trans isomers of N-[1,5-diphenyl-4-methyl-3-penten-2-yl]amine, respectively. These amines form via hydrolysis of N-formylamino derivatives generated from unsaturated ketone intermediates. These findings demonstrate that reaction temperature and time strongly govern both amphetamine formation and impurity composition. The identified impurities serve as distinct chemical indicators, enabling reconstruction of illicit synthesis conditions and supporting intelligence on amphetamine production.</p>","PeriodicalId":160,"journal":{"name":"Drug Testing and Analysis","volume":" ","pages":""},"PeriodicalIF":2.7,"publicationDate":"2026-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147571479","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}