Assessment of the stability of mephedrone in ante-mortem and post-mortem blood specimens

Busardò, Francesco Paolo; Kyriakou, Chrystalla; Tittarelli, Roberta; Mannocchi, Giulio; Pantano, Flaminia; Santurro, Alessandro; Zaami, Simona; Baglìo, Giovanni

doi:10.1016/j.forsciint.2015.07.021

Cited by 45 publications

(42 citation statements)

References 27 publications

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“…Often there is a delay between sample collection and analysis, and the best storage conditions have to be applied until analysis could be performed. Stability of synthetic cathinones in urine [3,10,11] and in blood/plasma [10,[12][13][14][15] has been reported; however, OF data is scarce [9].…”

Section: Discussionmentioning

confidence: 99%

Stability of synthetic cathinones in oral fluid samples

Miller

Kim

Concheiro

2017

Forensic Science International

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Synthetic cathinones are new stimulant drugs derived from cathinone that have been sold as "legal highs" worldwide. These compounds can elicit powerful effects such as delusions, hallucinations as well as other potentially dangerous behavior. New analogs with varying effects and potencies are constantly introduced in the market to evade legislation, and they are not detected by routine screening and confirmation methods. Oral fluid is an alternative matrix of increasing interest in forensic toxicology. Its collection is non-invasive and easily supervised, and positive drug findings typically reflect recent drug exposure. The focus of this research was to develop a method for the determination of 10 synthetic cathinones (cathinone, methcathinone, buphedrone, mephedrone, 4methylethcathinone, 3,4-methylenedioxypyrovalerone (MDPV), methylone, naphyrone, alpha-pyrrolidinovalerophenone (PVP) and N-ethylcathinone) in preserved oral fluid (Quantisal TM ), as well as evaluate their stability in preserved (Quantisal and Oral-Eze TM ) and neat oral fluid samples stored under different conditions, using ultrahigh-performance liquid chromatography-tandem mass spectrometry (UHPLC-MSMS). Four-hundred μL oral fluid-Quantisal buffer mixture (100 μL oral fluid and 300 μL buffer) were subjected to cation exchange solid phase extraction. The chromatographic reverse-phase separation was achieved with a gradient mobile phase of 0.1 % formic acid in water and in acetonitrile in 5 min. We used a Shimadzu triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode. The assay was linear from 1-250 ng/mL, with the limits of detection of 0.75-1 ng/mL. Imprecision (n=15) was <20.7% and accuracy (n=15) was 84-115.3%. Extraction efficiency was 87.2-116.8% (n=6), process efficiency was 30.9-103.7% (n=6), and matrix effect was -65.1 to -6.2% n=6). The stability was performed for neat oral fluid, oral fluid in Quantisal buffer, and oral fluid in Oral-Eze buffer samples stored up to one month at room temperature, 4 o C and -20 o C, and after 3 freezethaw cycles. Losses up to -71.2 to -100% were observed in neat and preserved samples stored at room temperature up to one month. At 4 o C, losses up to -88.2% occurred in neat OF and Oral-Eze samples, while Quantisal samples showed losses up to -34%. All types samples were stable if stored at -20 o C and after 3 freeze-thaw cycles.

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Section: Discussionmentioning

confidence: 99%

Stability of synthetic cathinones in oral fluid samples

Miller

Kim

Concheiro

2017

Forensic Science International

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“…The stability of mephedrone has previously been investigated in human whole blood containing different preservatives and stored under different conditions . Mephedrone has been reported to be most stable at −20°C when preserved with acidic preservatives (NaF/KOx and NaF/citrate buffer).…”

Section: Introductionmentioning

confidence: 99%

“…6 The stability of mephedrone has previously been investigated in human whole blood containing different preservatives and stored under different conditions. [10][11][12][13] Mephedrone has been reported to be most stable at −20°C when preserved with acidic preservatives (NaF/KOx and NaF/citrate buffer). The underlying cause of its instability in biological matrices is unknown but a previous study looking at mephedrone degradation in alkaline solution suggests the involvement of oxidants such as dissolved oxygen.…”

mentioning

confidence: 99%

Stability of mephedrone and five of its phase I metabolites in human whole blood

Czerwinska

Parkin

Dargan

et al. 2018

Drug Testing and Analysis

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Mephedrone is a new psychoactive substance known to be unstable in biological matrices stored at room temperature or refrigerated. While the instability of mephedrone has been investigated before, there is currently no data regarding the stability of mephedrone metabolites. In this study, a liquid chromatography–tandem mass spectrometry method for the simultaneous quantification of mephedrone and five of its phase I metabolites (dihydro‐mephedrone, nor‐mephedrone, hydroxytolyl‐mephedrone, 4‐carboxy‐mephedrone and dihydro‐nor‐mephedrone) in human whole blood has been developed and validated. Samples were extracted by a mixed mode solid‐phase extraction and analyzed on a pentafluorophenylpropyl column. The method was successfully validated for selectivity, linearity (0.2–2 to 10–100 ng/mL), limits of detection (50–500 pg/mL) and quantification (200–2000 pg/mL), precision (0.924–8.27%), accuracy (86.6–115%), carryover, recovery (32.5–88.3%), and matrix effects (71.0–108%). Analyte stability in human whole blood preserved with sodium fluoride/potassium oxalate was assessed at +4°C and −20°C after 24 hours, 48 hours, 4 days, and 10 days of storage. Instability was observed in samples stored at +4°C: nor‐mephedrone and 4‐carboxy‐mephedrone lost 40.2 ± 6.7% and 48.1 ± 4.8%, respectively, of their initial concentration at low concentration level and 33.8 ± 4.2% and 44.6 ± 6.5%, respectively, at high concentration level after 10 days. All analytes were more stable at −20°C where the highest loss of 22.6 ± 6.9% was observed for 4‐carboxy‐mephedrone after 10 days. This is the first time stability of mephedrone metabolites in human whole blood has been assessed, indicating −20°C to be the recommended storage condition for all analytes in clinical settings.

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“…In this phenomenon, the concentration of drugs in the blood and other tissues can change up to 15-fold after death 12 . It is thought that all drugs will be affected to some extent by postmortem redistribution and the main contributing factors are (1) the time between death and sampling, (2) the site(s) of sampling (with femoral blood being the sample that is least affected by postmortem redistribution), (3) potential postmortem metabolism/production by either body enzymes or bacteria and finally, (4) physicochemical properties of the drug (such as pKa, LogP and most importantly volume of distribution (Vd)) 2, 13 .…”

Section: Discussionmentioning

confidence: 99%

“…Besides to some users’ self-report euphoria and an increased perception and feeling of closeness to others, MDMA users may develop acute complications with potential fatal consequences when taken in warm environments 12 . As for corpse, the lower storage temperature stopped postmortem redistribution of MDMA partly at 0 °C and humidity of 50%, showing less change of concentrations than 20 °C and humidity of 70%, no matter in any sample.…”

Section: Discussionmentioning

confidence: 99%

How Postmortem Redistribution of MDMA in Acute Alcohol-MDMA Combined-Use Rats Change under Effects of Alcohol

Liang

Zhang

Zheng³

et al. 2017

Sci Rep

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MDMA is often taken recreationally with alcohol as combined-use. The objective was to determine MDMA postmortem redistribution (PMR) and corresponding effects in combined-style under different storage conditions. Steps were 20%-mixture of alcohol-water for initial 4 weeks to Group-A&B and intragastric infusions of MDMA (150 mg/kg) to Group-A later; in the same time, drinking pure water to Group-C&D first and then MDMA-fed to Group-C. The sacrificed rats were kept under different conditions for 10-d, during which the body fluids and tissues were collected on 15 continuous time-points and then detected. The MDMA concentrations were quite different along with postmortem interval (PMI) went by; the area under concentration-PMI curve significantly increased with combined-alcohol in comparison to MDMA alone, while that significantly decreased by lowering preservation temperature, allied with corresponding humidity. Combined-alcohol could exacerbate PMR of MDMA, as concentrations of combined-use rats’ samples were quite higher than mono-MDMA ones under any conditions, while different for body fluids and tissues; meanwhile lowering storage temperature could alleviate effects of alcohol. The study implies that in case of combined-use, the changes of concentrations are probably effected by some combined component, especially when come to identification of toxic level or even death.

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Assessment of the stability of mephedrone in ante-mortem and post-mortem blood specimens

Cited by 45 publications

References 27 publications

Stability of synthetic cathinones in oral fluid samples

Stability of synthetic cathinones in oral fluid samples

Stability of mephedrone and five of its phase I metabolites in human whole blood

How Postmortem Redistribution of MDMA in Acute Alcohol-MDMA Combined-Use Rats Change under Effects of Alcohol

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