Quantitation of Fentanyl and Metabolites from Liver Tissue Using a Validated QuEChERS Extraction and LC–MS-MS Analysis

2020 ◽  
Author(s):  
Joseph Cox ◽  
Alex Train ◽  
Avery Field ◽  
Colby Ott ◽  
Joseph DelTondo ◽  
...  
Keyword(s):  
Matrix Effects ◽  
Linear Range ◽  
Mass Spectrometry Analysis ◽  
Forensic Sciences ◽  
Opioid Overdose ◽  
Spectrometry Analysis ◽  
Analytical Scheme ◽  
Target Drug ◽  
Ionization Suppression ◽  
High Concentrations

Abstract According to the National Institute on Drug Abuse (NIDA), more than one hundred people die every day from opioid overdose. Overdose fatalities have risen as the availability of potent synthetic opioids, such as fentanyl, has increased. A forensic postmortem toxicological specimen is often in various stages of decomposition, experiencing autolysis and putrefaction, which complicates the extraction, creating a difficult challenge for toxicologists. Isolating the target drug, while creating an efficient and simplified analytical scheme, is a goal for most toxicology laboratories. The validation of a quick, easy, cheap, effective, rugged and safe extraction protocol is presented in this study as an alternative analytical method for efficient extraction and detection of fentanyl and its major metabolites: norfentanyl and despropionyl fentanyl (4-ANPP). The liquid Chromatography with tandem mass spectrometry analysis was validated following the American Academy of Forensic Sciences Standards Board (ASB) standard 036 proposed requirements. Evaluated parameters include selectivity, matrix effects (MEs), linearity, processed sample stability, bias, precision and proof of applicability using liver samples from authentic postmortem cases. MEs (represented as percent ionization suppression or enhancement) at low and high concentrations were −10.0% and 1.4% for fentanyl, −2.1% and −0.3% for 4-ANPP and 3.1% and 2.8% for norfentanyl, respectively. Bias for the three analytes ranged from −8.5% to −19.9% for the low concentrations, −3.6% to −14.7% for the medium concentrations and 1.5% to −16.1% for the high concentrations with all being within the ±20% guideline. Precision for the three analytes ranged from 2.2% to 15.1%. The linear range for the fentanyl and norfentanyl was 0.5–100 and 4-ANPP had a linear range of 0.4–80 μg/kg. The authentic postmortem liver samples ranged in fentanyl concentrations from 56.6 to 462.3 μg/kg with a mean of 149.2 μg/kg (n = 10). The range of norfentanyl concentrations were 1.9 to 50.0 μg/kg with a mean of 14.1 μg/kg (n = 10). The range of 4-ANPP concentrations were 3.2 to 23.7 μg/kg with a mean of 7.5 μg/kg (n = 7).

Quantitation and Validation of 34 Fentanyl Analogs from Liver Tissue Using a QuEChERS Extraction and LC–MS-MS Analysis

2021 ◽  
Author(s):  
Joseph Cox ◽  
Kylea Mathison ◽  
Colby Ott ◽  
Joseph DelTondo ◽  
James C Kraner ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liver Tissue ◽  
Mass Spectrometry Analysis ◽  
Forensic Sciences ◽  
High Concentration ◽  
Spectrometry Analysis ◽  
Fentanyl Analogs ◽  
Ionization Suppression ◽  
High Concentrations ◽  
Overdose Deaths

Abstract Since 2013, drug overdose deaths involving synthetic opioids (including fentanyl and fentanyl analogs) have increased from 3,105 to 31,335 in 2018. Postmortem toxicological analysis in fentanyl-related overdose deaths is complicated by the high potency of the drug, often resulting in low analyte concentrations and associations with toxicity, multidrug use, novelty of emerging fentanyl analogs and postmortem redistribution. Objectives for this study include the development of a quick, easy, cheap, effective, rugged and safe (QuEChERS) extraction and subsequent liquid chromatography–mass spectrometry/mass spectrometry analysis, validation of the method following the American Academy of Forensic Sciences Standards Board (ASB) standard 036 requirements and application to authentic liver specimens for 34 analytes including fentanyl, metabolites and fentanyl analogs. The bias for all 34 fentanyl analogs did not exceed ±10% for any of the low, medium or high concentrations and the %CV did not exceed 20%. No interferences were identified. All 34 analytes were within the criteria for acceptable percent ionization suppression or enhancement with the low concentration ranging from −10.2% to 23.7% and the high concentration ranging from −7.1% to 11.0%. Liver specimens from 22 authentic postmortem cases were extracted and analyzed with all samples being positive for at least one target analyte from the 34 compounds. Of the 22 samples, 17 contained fentanyl and metabolites plus at least one fentanyl analog. The highest concentration for a fentanyl analog was 541.4 μg/kg of para-fluoroisobutyryl fentanyl (FIBF). The concentrations for fentanyl (n = 20) ranged between 3.6 and 164.9 μg/kg with a mean of 54.7 μg/kg. The fentanyl analog that was most encountered was methoxyacetyl fentanyl (n = 11) with a range of 0.2–4.6 μg/kg and a mean of 1.3 μg/kg. The QuEChERS extraction was fully validated using the ASB Standard 036 requirements for fentanyl, metabolites and fentanyl analogs in liver tissue.

Molecular Changes in Dissolved Organic Matter After Soil Rewetting

2021 ◽  
Author(s):  
Alice Orme ◽  
Simon Benk ◽  
Markus Lange ◽  
Christian Zerfaß ◽  
Georg Pohnert ◽  
...  
Keyword(s):  
Organic Matter ◽  
Dissolved Organic Matter ◽  
Mass Spectrometry Analysis ◽  
Microbial Decomposition ◽  
Molecular Changes ◽  
Spectrometry Analysis ◽  
Subsurface Waters ◽  
High Production ◽  
High Concentrations ◽  
Over Time

<p>The intensity and occurrence of droughts is projected to increase due to climate change. Dried soils release high concentrations of dissolved organic matter (DOM) into subsurface waters when they are rewet, the so-called rewetting peak. To more accurately predict the role of rewetting of soils after drought on the carbon cycle in a changing climate, it is important to understand the processes behind this DOM release.</p><p>The DOM rewetting peak origin is disputed between soil organic matter (SOM) from breakdown of soil particles; accumulated root exudates; and microbial release due to a change in osmotic potential through osmolytes or cell bursting. To better understand the origin of the rewetting DOM peak, we took a rewetting series of soil water samples from different vegetation types between December 2018 and April 2019 for targeted and untargeted metabolomics. Initial results using untargeted ultrahigh-resolution mass spectrometry analysis revealed a clear temporal trend, indicating that vegetation-independent molecular changes occur following rewetting. An increase in O/C and a decrease in H/C over time was observed which is attributed to microbial decomposition, supported by a decrease in m/z over time. We also observed an increase in the content of lipidic compounds (R > 0.6) following rewetting. This indicates that cells do not burst upon rewetting and, over time, microbial activity increases, suggesting that the DOM rewetting peak is caused by a lack of decomposition, rather than a high production, of organic matter.</p>

A novel Sphingobacterium sp. RB, a rhizosphere isolate degrading para-nitrophenol with substrate specificity towards nitrotoluenes and nitroanilines

2019 ◽  
Vol 366 (14) ◽  
Author(s):  
Rachel Samson ◽  
Ragini Bodade ◽  
Smita Zinjarde ◽  
Razia Kutty
Keyword(s):  
Sole Source ◽  
Nitroaromatic Compounds ◽  
Mass Spectrometry Analysis ◽  
Liquid Chromatography Mass Spectrometry ◽  
Spectrometry Analysis ◽  
Morphological Variations ◽  
Microscope Images ◽  
High Concentrations ◽  
Bioremediation Agent ◽  
Scanning Electron

ABSTRACT Sphingobacterium sp. RB, a novel bacterial strain isolated from a soil sample, was able to utilize para-nitrophenol (PNP) as sole source of carbon and energy at high concentrations (1.0–5.0 mM). The culture completely degraded 3.0 mM PNP within 36 h with proportionate increase in biomass. With 5.0 mM PNP (700 ppm), 70% degradation was observed within 72 h of incubation. Scanning electron microscope images of the isolate in the presence and absence of PNP showed no significant morphological variations. Liquid chromatography–mass spectrometry analysis indicated that the biodegradation of PNP in this bacterium proceeded via the formation of 1,2,4-benzenetriol. Cells previously exposed to PNP (induced) were 30% more effective in degrading PNP. With catechol and phenol, such induction was not observed. Uninduced cells of Sphingobacterium sp. RB were capable of degrading a variety of other nitroaromatic compounds, including 2-nitroaniline, 2,4-dinitroaniline, 2-nitrotoluene, 3-nitrotoluene and 2,4-dinitrophenol, within 72 h, thus proving its candidacy as a potent bioremediation agent. To the best of our knowledge, this is the first report on a Sphingobacterium species degrading PNP via formation of 1,2,4-benzenetriol.

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