RESUMEN
Endogenous electrophiles, ionizing and non-ionizing radiation, and hazardous chemicals present in the environment and diet can damage DNA by forming covalent adducts. DNA adducts can form in critical cancer driver genes and, if not repaired, may induce mutations during cell division, potentially leading to the onset of cancer. The detection and quantification of specific DNA adducts are some of the first steps in studying their role in carcinogenesis, the physiological conditions that lead to their production, and the risk assessment of exposure to specific genotoxic chemicals. Hundreds of different DNA adducts have been reported in the literature, and there is a critical need to establish a DNA adduct mass spectral database to facilitate the detection of previously observed DNA adducts and characterize newly discovered DNA adducts. We have collected synthetic DNA adduct standards from the research community, acquired MSn (n = 2, 3) fragmentation spectra using Orbitrap and Quadrupole-Time-of-Flight (Q-TOF) MS instrumentation, processed the spectral data and incorporated it into the MassBank of North America (MoNA) database, and created a DNA adduct portal Web site (https://sites.google.com/umn.edu/dnaadductportal) to serve as a central location for the DNA adduct mass spectra and metadata, including the spectral database downloadable in different formats. This spectral library should prove to be a valuable resource for the DNA adductomics community, accelerating research and improving our understanding of the role of DNA adducts in disease.
Asunto(s)
Aductos de ADN , ADN , Humanos , ADN/química , Espectrometría de Masas , Daño del ADN , CarcinogénesisRESUMEN
Currently, there is a significant demand in forensic toxicology for biomarkers of cannabis exposure that, unlike ∆9-tetrahydrocannabinol, can reliably indicate time and frequency of use, be sampled with relative ease, and correlate with impairment. Oral fluid (OF) and exhaled breath condensate (EBC) are alternative, non-invasive sample matrices that hold promise for identifying cannabis exposure biomarkers. OF, produced by salivary glands, is increasingly utilized in drug screening due to its non-invasive collection and is being explored as an alternative matrix for cannabinoid analysis. EBC is an aqueous specimen consisting of condensed water vapor containing water-soluble volatile and non-volatile components present in exhaled breath. Despite potential advantages, there are no reports on the use of EBC for cannabinoid detection. This study developed a supported liquid extraction approach and LC-QqQ-MS dMRM analytical method for quantification of 25 major and minor cannabinoids and metabolites in OF and EBC. The method was validated according to the ANSI/ASB 036 standard and other published guidelines. LOQ ranged from 0.5 to 6.0 ng/mL for all cannabinoids in both matrices. Recoveries for most analytes were 60-90%, with generally higher values for EBC compared to OF. Matrix effects were observed with some cannabinoids, with effects mitigated by use of matrix-matched calibration. Bias and precision were within ± 25%. Method applicability was demonstrated by analyzing ten authentic OF and EBC samples, with positive detections of multiple analytes in both matrices. The method will facilitate comprehensive analysis of cannabinoids in non-invasive sample matrices for the development of reliable cannabis exposure biomarkers.
Asunto(s)
Pruebas Respiratorias , Cannabinoides , Límite de Detección , Saliva , Cannabinoides/análisis , Pruebas Respiratorias/métodos , Humanos , Saliva/química , Detección de Abuso de Sustancias/métodos , Cromatografía Liquida/métodos , Espectrometría de Masas en Tándem/métodos , Espiración , Reproducibilidad de los ResultadosRESUMEN
Aptamers are promising biorecognition elements for sensors. However, aptamer-based assays often lack the requisite levels of sensitivity and/or selectivity because they typically employ structure-switching aptamers with attenuated affinity and/or utilize reporters that require aptamer labeling or which are susceptible to false positives. Dye-displacement assays offer a label-free, sensitive means for overcoming these issues, wherein target binding liberates a dye that is complexed with the aptamer, producing an optical readout. However, broad utilization of these assays has been limited. Here, we demonstrate a rational approach to develop colorimetric cyanine dye-displacement assays that can be broadly applied to DNA aptamers regardless of their structure, sequence, affinity, or the physicochemical properties of their targets. Our approach should accelerate the development of mix-and-measure assays that could be applied for diverse analytical applications.
RESUMEN
Mass spectrometry-based DNA adductomics is an emerging approach for the human biomonitoring of hazardous chemicals. A mass spectral database of DNA adducts will be created for the scientific community to investigate the associations between chemical exposures, DNA damage, and disease risk.
Asunto(s)
Aductos de ADN/efectos de los fármacos , Bases de Datos de Compuestos Químicos , Contaminantes Ambientales/farmacología , Compuestos Orgánicos/farmacología , Daño del ADN , Contaminantes Ambientales/química , Humanos , Espectrometría de Masas , Compuestos Orgánicos/químicaRESUMEN
The class of novel psychoactive substances known as synthetic cannabinoids (SC) includes illicit compounds that are sprayed on plant material and smoked or sold as liquids to be vaporized in e-cigarettes. In toxicological analysis of SC, fast analytical methods are needed for the detection and confirmation of parent drugs and metabolites at very low levels. While various analytical methods have been developed for SC in blood and urine, few are available for alternative matrices such as oral fluid (OF). There are numerous advantages to using OF as a sample matrix for SC analysis, including non-invasive collection, lesser risk of adulteration, and presence of both parent drug and metabolites. Here we report a validated online solid-phase extraction (online SPE) method coupled to LC-QqQ-MS for rapid confirmation and quantitation of 72 structurally diverse SC parent drugs and metabolites in OF with 2.5 min of preconcentration time and a total elution time of < 10 min. The use of online SPE for sample pretreatment facilitates rapid and consistent processing and greatly increases sample throughput. The method was fully validated according to relevant guidelines (ANSI/ASB Standard 036). Bias and precision values were within ± 20% for all compounds in human OF matrix. Method detection and quantitation limits ranged from 0.4 to 3.8 ng/mL and from 1.1 to 11.6 ng/mL, respectively. Recovery, matrix effects, process efficiency, carryover, and stability were also within acceptable limits for the majority of compounds. Successful application of the method was demonstrated using blank human OF fortified with SC in addition to a set of authentic OF specimens previously tested by another laboratory. Graphical abstract.
Asunto(s)
Cannabinoides/metabolismo , Cromatografía Liquida/métodos , Drogas Ilícitas/análisis , Espectrometría de Masas/métodos , Saliva/metabolismo , Extracción en Fase Sólida/métodos , Humanos , Límite de Detección , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
A rapid method for the characterization of both organic and inorganic components of gunshot residues (GSR) is proposed as an alternative tool to facilitate the identification of a suspected shooter. In this study, two fast screening methods were developed and optimized for the detection of organic compounds and inorganic components indicative of GSR presence on the hands of shooters and non-shooters. The proposed methods consist of headspace extraction of volatile organic compounds using a capillary microextraction of volatiles (CMV) device previously reported as a high-efficiency sampler followed by detection by GC-MS. This novel sampling technique has the potential to yield fast results (<2min sampling) and high sensitivity capable of detecting 3ng of diphenylamine (DPA) and 8ng of nitroglycerine (NG). Direct analysis of the headspace of over 50 swabs collected from the hands of suspected shooters (and non-shooters) provides information regarding VOCs present on their hands. In addition, a fast laser induced breakdown spectroscopy (LIBS) screening method for the detection of the inorganic components indicative of the presence of GSR (Sb, Pb and Ba) is described. The sampling method for the inorganics consists of liquid extraction of the target elements from the same cotton swabs (previously analyzed for VOCs) and an additional 30 swab samples followed by spiking 1µL of the extract solution onto a Teflon disk and then analyzed by LIBS. Advantages of LIBS include fast analysis (~12s per sample) and high selectivity and sensitivity, with expected LODs 0.1-18ng for each of the target elements after sampling. The analytical performance of the LIBS method is also compared to previously reported methods (inductively coupled plasma-optical emission spectroscopy). The combination of fast CMV sampling, unambiguous organic compound identification with GC-MS and fast LIBS analysis provides the basis for a new comprehensive screening method for GSR.
RESUMEN
Elemental analysis of glass was conducted by 16 forensic science laboratories, providing a direct comparison between three analytical methods [micro-x-ray fluorescence spectroscopy (µ-XRF), solution analysis using inductively coupled plasma mass spectrometry (ICP-MS), and laser ablation inductively coupled plasma mass spectrometry]. Interlaboratory studies using glass standard reference materials and other glass samples were designed to (a) evaluate the analytical performance between different laboratories using the same method, (b) evaluate the analytical performance of the different methods, (c) evaluate the capabilities of the methods to correctly associate glass that originated from the same source and to correctly discriminate glass samples that do not share the same source, and (d) standardize the methods of analysis and interpretation of results. Reference materials NIST 612, NIST 1831, FGS 1, and FGS 2 were employed to cross-validate these sensitive techniques and to optimize and standardize the analytical protocols. The resulting figures of merit for the ICP-MS methods include repeatability better than 5% RSD, reproducibility between laboratories better than 10% RSD, bias better than 10%, and limits of detection between 0.03 and 9 µg g(-1) for the majority of the elements monitored. The figures of merit for the µ-XRF methods include repeatability better than 11% RSD, reproducibility between laboratories after normalization of the data better than 16% RSD, and limits of detection between 5.8 and 7,400 µg g(-1). The results from this study also compare the analytical performance of different forensic science laboratories conducting elemental analysis of glass evidence fragments using the three analytical methods.
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Compared with other racial/ethnic groups in the United States (US), American Indians/Alaska Natives have one of the fastest climbing rates of drug overdose deaths involving stimulants. Validating the substances self-reported by Indigenous people who use injection drugs (IPWIDs) can present logistical and cultural challenges. While the collection of biospecimens (e.g., urine, blood, hair follicle) can be one way to cross-validate the substances self-reported by IPWIDs, the collection of biospecimens has been historically problematic when conducting substance use research with Indigenous North Americans. In our National Institutes of Health (NIH)-supported pilot research conducted with IPWIDs, we have documented low willingness to provide a biospecimen to a research team. This article demonstrates an alternative method for validating self-reported substances injected by IPWIDs that does not require the extraction of biospecimens from Indigenous bodies and spaces. The method described includes:â¢Collecting used, unwashed syringes from IPWIDs at the time of behavioral assessment,â¢Sampling the used syringe by washing the syringe needle/barrel with methanol,â¢Analyzing the samples with gas chromatography mass spectrometry (GC-MS) and liquid chromatography coupled to triple-quadrupole mass spectrometry (LC-QQQ-MS). This method offers a more culturally appropriate alternative to validate substances self-reported by IPWIDs during behavioral assessments.
RESUMEN
A novel phenyl modified PDMS (PhPDMS) sol-gel adsorption phase was developed for use with the capillary microextraction of volatiles (CMV) device, and determined to provide significant enhancement in BTEX recoveries when sampling trace (ng) amounts of these volatiles at ambient conditions. The previously reported reusable PDMS-CMV device has been demonstrated to rapidly and efficiently extract target compound's vapors in forensic and environmental applications. An improved recovery for VOCs was achieved with a cryofocusing system while extracting at -10⯰C, but it was found to be impractical for field sampling. This report details a modification to the CMV's chemistry, by the successful introduction of phenyl groups to the PDMS sol-gel adsorption phase, allowing enhanced performance at ambient extraction conditions. Higher average recoveries, determined through a broad concentration range, were demonstrated for PhPDMS-CMV over its original PDMS-CMV, from cans simulating a closed space set-up. Within 7.8 (±10%) and 3.5 (±6%) folds higher for benzene and toluene, respectively and 2 (±2%) folds for ethylbenzene and xylenes. Significant higher retaining capabilities were demonstrated also at the more challenging set-up, simulating an open space environment. Whereas, benzene had completely breakthrough the PDMS-CMV, its reliable detection was still confirmed with PhPDMS-CMV pumping at 2â¯L or 6â¯L air, concentration dependent. At least 50 folds (±26%) more toluene was retained with PhPDMS-CMV at 6â¯L air than with PDMS-CMV. The enhanced overall performance lead to determination of trace LODs with the new CMV of 0.002, 0.00035 and 0.00015â¯ppm for benzene, toluene, ethyl benzene and xylenes, respectively. As proof of concept, for the first time solvent extraction is presented for the new CMV as an alternative to thermal desorption extraction. Extraction efficiencies of 60% for TEX, and lower concentration dependent for benzene, were demonstrated with the ease and rapid application of 100⯵L acetone through the device. The improvements described in this study continues to build on the potential for the use of the reusable new CMV device by expanding its possible potential applications for fast and sensitive air sampling of VOCs. The solvent extraction step may offer compatibility with LC-based systems.