Quantitative Estimation of 38 Illicit Psychostimulants in Blood by GC-APCI-QTOFMS with Nitrogen Chemiluminescence Detection Based on Three External Calibrators

A method was developed for quantitative estimation of illicit psychostimulants in blood, with an emphasis on new psychoactive substances, based on gas chromatography nitrogen chemiluminescence detection coupled with atmospheric pressure chemical ionization quadrupole time-of-flight mass spectrometry (GC-NCD-APCI-QTOFMS). Quantitative estimation relied on the NCD's N-equimolar response to nitrogen, using amphetamine, 3,4-methylenedioxymethamphetamine (MDMA) and methylenedioxypyrovalerone as external calibrators for prim-, sec- and tert- amines, respectively. After spiking with 38 stimulants at 3 concentration levels, the donor blood samples were submitted to liquid-liquid extraction at a basic pH followed by acylation with trifluoroacetic anhydride. All but 3 psychostimulants could be analyzed with a limit of quantification (LOQ) of 0.05 mg/L. At LOQ, the coefficient of variation (CV) values for between-day accuracy was 62.3-143.3% (mean, 93.5%; median, 88.5%) and precision 6.6-22.4% (mean, 15.8%; median, 16.1%). In addition, 11 post-mortem blood samples, containing 0.08-2.4 mg/L of amphetamine (n = 5), methamphetamine (n = 4) or MDMA (n = 4), were analyzed by the GC-NCD-APCI-QTOFMS method, and the results were compared with an established electron ionization GC-MS method with appropriate calibration. The agreement between the 2 methods was 62.5-117.3%. Regarding identification, the APCI source permitted detection of the intact precursor ion, or the respective acylation product, for all of the measured compounds. The GC-NCD-APCI-QTOFMS method developed here enables instant quantitative estimation of illicit psychostimulants in blood at reasonable accuracy, without the necessity of possessing the true reference standards for each analyte.Peer reviewe

Toxic lifespan of the synthetic opioid U-47,700 in Finland verified by re-analysis of UPLC-TOF-MS data

U-47,700 is a synthetic opioid that emerged on the novel psychoactive substance market a few years ago. After incorporating the substance into the urine UPLC-TOF-MS screening used in post-mortem toxicology, the drug was detected in 10 autopsy cases within routine case work. In all cases, the cause of death was accidental poisoning by U-47,700 alone or in combination with other psychoactive substances. The concentration of U-47,700 in the blood samples ranged between 0.15-2.0 mg/L with a median of 0.30 mg/L. In one of the cases with a U-47,700 concentration of 0.27 mg/L, no other psychoactive substances were detected. The stored TOF-MS analytical data from the year preceding the incorporation of U-47,700 into the screening was reprocessed in order to search for more positive cases. The data-independent acquisition of the original screening allowed for retrospective re-analysis of the full-scan data without additional experiments on the actual sample. The retrospective data-analysis revealed two additional cases positive for U-47,700. The first mention of U-47,700 on a Finnish internet discussion forum was in March 2015. After having been detected in several death cases, the drug was put under national control in November 2016 and the last fatality occurred in 2017. The toxic lifespan of U-47,700 thus lasted for approximately 2 years in Finland. Forensic and clinical laboratories need to rapidly adjust their screening procedures in order to adapt to the continuously expanding field of novel psychoactive substances. Retrospective data-analysis is a practical tool for monitoring the emergence of new substances onto the market. (C) 2019 Elsevier B.V. All rights reserved.Peer reviewe

Development of a GC-APCI-QTOFMS library for new psychoactive substances and comparison to a commercial ESI library

Gas chromatography coupled to atmospheric pressure chemical ionization quadrupole time-of-flight mass spectrometry (GC-APCI-QTOFMS) was evaluated for the identification of new psychoactive substances (NPS). An in-house high mass resolution GC-APCI-QTOFMS test library was developed for 29 nitrogen-containing drugs belonging mostly to synthetic stimulants. The library was based on 12 intra-day measurements of each compound at three different collision energies, 10, 20 and 40 eV. The in-house library mass spectra were compared to mass spectra from a commercial library constructed by liquid chromatography-electrospray ionization (LC-ESI) QTOFMS. The reversed library search scores between the in-house GC-APCI library and the commercial LC-ESI library were compared once a week during a 5-week period by using data measured by GC-APCI-QTOFMS. The protonated molecule was found for all drugs in the full scan mode, and the drugs were successfully identified by both libraries in the targeted MS/MS mode. The GC-APCI library score averaged over all collision energies was as high as 94.4/100 with a high repeatability, while the LC-ESI library score was also high (89.7/100) with a repeatability only slightly worse. These results highlight the merits of GC-APCI-QTOFMS in the analysis of NPS even in situations where the reference standards are not immediately available, taking advantage of the accurate mass measurement of the protonated molecule and product ions, and comparison to existing soft-ionization mass spectral libraries.Peer reviewe

Biological variation of biochemical urine and serum analytes in healthy dogs

BackgroundBiological variation (BV) of urinary (U) biochemical analytes has not been described in absolute terms, let alone as a ratio of the U-creatinine or fractional excretion in healthy dogs. These analytes are potential diagnostic tools for different types of kidney damage and electrolyte disorders in dogs. ObjectivesWe aimed to investigate the BV of specific gravity, osmolality, creatinine, urea, protein, glucose, chloride, sodium, potassium, calcium, and phosphate in urine from healthy pet dogs. MethodsBlood and urine samples from 13 dogs were collected once weekly for 8 weeks. Samples were analyzed in duplicate and in randomized order. For each sample, U-analyte and serum concentrations were measured, and U-analyte/U-creatinine and fractional excretion (FE) were calculated. Components of variance, estimated by restricted maximum likelihood, were used to determine within-subject variation (CVI), between-subject variation (CVG), and analytical variation (CVA). Index of individuality (II) and reference change values were calculated. ResultsCV(I) for all urine analytes varied between 12.6% and 35.9%, except for U-sodium, U-sodium/U-Cr, and FE-sodium, which had higher CV(I)s (59.5%-60.7%). For U-protein, U-sodium, U-potassium, U-sodium/U-creatinine, FE-urea, FE-glucose, FE-sodium, FE-potassium, and FE-phosphate II were low, indicating that population-based RIs were appropriate. The remaining analytes had an intermediate II, suggesting that population-based RIs should be used with caution. ConclusionThis study presents information on the biological variation of urinary and serum biochemical analytes from healthy dogs. These data are important for an appropriate interpretation of laboratory results

Mitochondrial respiratory dysfunction due to the conversion of substituted cathinones to methylbenzamides in SH-SY5Y cells

The increased use of cathinone-type designer drugs, known as legal highs, has led to concerns about their potential neurotoxicity due to their similarity to methamphetamine (METH). Therefore, closer investigations of their toxic effects are needed. We investigated the effects of the cathinones 4-methylmethcathinone (4-MMC) and 3,4-methylenedioxymethcathinone (MDMC) and the amphetamine METH on cytotoxicity and mitochondrial respiration in SH-SY5Y neuroblastoma cells. We also investigated the contribution of reactive species, dopamine, Bcl-2 and tumor necrosis factor alpha(TNF alpha) on toxicity. Finally, we investigated the effect of cathinone breakdown products using ultra-high performance liquid chromatography/high-resolution time-of-flight mass spectrometry and studied their involvement in toxicity. We observed dose-dependent increases in cytotoxicity and decreases in mitochondrial respiration following treatment with all cathinones and amphetamines. Glutathione depletion increases amphetamine, but not cathinone toxicity. Bcl-2 and TNF alpha pathways are involved in toxicity but dopamine levels are not. We also show that cathinones, but not amphetamines, spontaneously produce reactive species and cytotoxic methylbenzamide breakdown products when in aqueous solution. These results provide an important first insight into the mechanisms of cathinone cytotoxicity and pave the way for further studies on cathinone toxicity in vivo.Peer reviewe

Simultaneous identification and quantification of new psychoactive substances in blood by GC-APCI-QTOFMS coupled to nitrogen chemiluminescence detection without authentic reference standards

A novel platform is introduced for simultaneous identification and quantification of new psychoactive substances (NPS) in blood matrix, without the necessity of using authentic reference standards. The instrumentation consisted of gas chromatography (GC) coupled to nitrogen chemiluminescence detection (NCD) and atmospheric pressure chemical ionization quadrupole time-of-flight mass spectrometry (APCI-QTOFMS). In this concept, the GC flow is divided in appropriate proportions between NCD for single-calibrant quantification, utilizing the detector's equimolar response to nitrogen, and QTOFMS for accurate mass-based identification. The principle was proven by analyzing five NPS, bupropion, desoxypipradrol (2-DPMP), mephedrone, methylone, and naphyrone, in sheep blood. The samples were spiked with the analytes post-extraction to avoid recovery considerations at this point. All the NPS studies produced a protonated molecule in APCI resulting in predictable fragmentation with high mass accuracy. The N-equimolarity of quantification by NCD was investigated by using external calibration with the secondary standard caffeine at five concentration levels between 0.17 and 1.7 mg/L in blood matrix as five replicates. The equimolarity was on average 98.7 %, and the range of individual equimolarity determinations was 76.7-130.1 %. The current analysis platform affords a promising approach to instant simultaneous qualitative and quantitative analysis of drugs in the absence of authentic reference standards, not only in forensic and clinical toxicology but also in other bioanalytical applications.Peer reviewe

Videomateriaali potilasohjauksen tukena IVF-hoidoissa

Ella kävi puolisonsa kanssa lapsettomuusklinikalla. Heillä on pää pyörällä, sillä lääkäri ja hoitajat käyttivät paljon erikoissanastoa. Lääkäri kertoi vastaanotolla, että päädytään IVF-hoitoon, joka aloitetaan follikkelien kypsytyksellä FSH-injektioiden avulla. Hän selitti mitä IVF-hoito on, mutta Ellaa mietitytti, miten follikkeleja kypsytetään ja miten lääkkeitä käytetään

Piirtäminen lapsen ilmaisukeinona

Miltä sinusta tuntuisi, jos muut eivät ymmärtäisi kieltäsi? Miten voimme havaita lapsen ahdistuneisuutta, jos lapsi ei osaa siitä kertoa? Lapsella voi olla paljon sellaista tietoa, mitä meidän aikuisten olisi hyvä kuulla. Tunteiden ilmaiseminen verbaalisesti voi olla kuitenkin vaikeaa ja tutkimus aiheestakin on vasta alussa. Hoitotyön opinnäytetyössä analysoitiin lasten ahdistuneisuuden tasoa piirustuksista