Search Results (8)

Green Analytical Chemistry (GAC) provides a framework for reducing hazardous reagents, energy consumption, and waste. The topic has gained momentum across many chemical industries over the past 25 years; however, progress in implementing sustainable methods and conducting greenness assessments within forensic laboratories has been comparatively slow. The purpose of this review is to highlight green approaches to analytical separation methods, including greenness assessment metrics, that have been reported in the literature for forensic chemistry and toxicology applications and to raise awareness of GAC in the forensic field. Recent scientific literature highlights promising advances in greener sample preparation and chromatographic approaches, particularly in forensic toxicology and seized-drug analysis. Emerging trends include the use of green solvents, bio-based and deep eutectic solvent systems, and the rapid expansion of microextraction techniques such as SPME, LPME, MEPS, FPSE, and DLLME, which reduce solvent volumes, minimize waste, and support higher-throughput workflows. Parallel developments in portable and miniaturized chromatographic instrumentation such as miniaturized LC–MS systems with increased detection specificity and Lab-on-a-Chip applications show promise for in situ measurements in the field. Ambient ionization mass spectrometry—in particular, DESI and DART—has had a major impact on forensic chemistry by providing tools for the rapid and direct analysis of chemical compounds in complex matrices with little or no sample preparation. Greenness assessment tools—including AGREE, AGREEprep, Eco-Scale, GAPI, and BAGI—are increasingly applied to evaluate analytical methods in forensic chemistry and toxicology, including those used for novel psychoactive substances. Although many green methodologies are well documented, their routine implementation remains limited. The continued integration of green solvents, microextractions, portable instrumentation, and standardized greenness metrics will be essential for advancing sustainable forensic separations. Full article

Polyfluoroalkyl substances (PFASs) and para-phenylenediamines (PPDs) are emerging classes of anthropogenic contaminants that are environmentally persistent (most often found in ground and surface water sources), bioaccumulative, and harmful to human health. These chemicals are currently regulated in the US by the Environmental Protection Agency (EPA), the Food and Drug Administration (FDA), and the Occupational Safety and Health Administration (OSHA). Analysis of these contaminants is currently spearheaded by mass spectrometry (MS) coupled to liquid chromatography (LC) because of their high sensitivity and separation capabilities. Although effective, a major flaw in LC-MS analysis is its large consumption of solvents and the amount of time required for each experiment. Direct analysis in real time mass spectrometry (DART-MS) is a new technique that offers high sensitivity and permits rapid analysis with little to no sample preparation. Herein, we present the qualitative and quantitative analysis of PFASs and PPDs by high-resolution DART-MS, interfaced with ion mobility (IM) and tandem mass spectrometry (MS/MS) characterization, demonstrating the utility of this multidimensional approach for the fast separation and detection of environmental contaminants. Full article

Saffron is a high-cost spice due to the specific conditions for optimal growth and because of being harvested by hand. The massive income from commercializing saffron substituted with other plant parts or low-cost spices makes this spice the main target of fraudsters. Background: Different methods have been developed for detecting saffron adulteration. Most of them are time consuming and complex, and in some types of analysis, the whole untargeted dataset is combined with advanced chemometric tools to differentiate authentic from non-authentic saffron. The official method, combining UV–vis spectroscopy and LC to determine the colour strength and the crocin content, is unable to detect saffron adulterants (safflower, marigold, or turmeric) added at a level lower than 20% (w/w). As a result, innovative approaches based on rapid, high-throughput methods for the identification of adulterated saffron samples are urgently demanded to counteract food frauds. Methods: This paper describes, for the first time, the development of a method combining Direct Analysis in Real Time (DART) with the triple quadrupole MS EVOQ based on the detection of specific MS/MS transitions, promoting a rapid, robust and chromatography-free method capable of monitoring safflower and turmeric adulteration in saffron. Results: The method proved to reach low LODs, allowing the determination of tiny amounts of turmeric and safflower powder in saffron as low as 3% and 5%, respectively, speeding up the whole analytical workflow and enabling us to perform 20 analyses in 10 min. Finally, the greenness of the method was also assessed according to the 0.88 score achieved by submitting it to the greenness calculator AGREE. Conclusions: Given its speed, simplicity, and robustness, this method stands out as a strong candidate for routine implementation in testing laboratories as a rapid screening tool to detect saffron adulteration with safflower or turmeric. Full article

Geniposidic 4-isoamyl ester (GENI) with anti-aging effects is a new iridoid glycoside derivative from Gardenia jasminoides Ellis found in our previous study. In this study, to indicate whether this compound has anti-Alzheimer’s disease (AD) effect, the galactose-induced AD mice and naturally aging mice with AD were used to do drug efficacy evaluation. Furthermore, the Western blot, small interfering RNA (siRNA), drug affinity responsive target stability (DARTS), cellular thermal shift assay (CESTA), liquid chromatography-tandem mass spectrometry (LC/MS-MS), adenosine 5′-monophosphate-activated protein kinase (AMPK) mutants and surface plasmon resonance (SPR) analysis were utilized to clarify the mechanism of action and identify target protein of this molecule. GENI exerts anti-AD efficacy in galactose-induced AD mice and naturally aging mice with AD through neuroprotection and modification of autophagy and neuron inflammation. Moreover, AMPK as the target protein of GENI to produce an anti-AD effect is identified and the ASP148, ASP157, and ASP166 of the AMPK α subunit and lysine (LYS)148, aspartic acid (ASP)156, LYS309, and ASP316 in the AMPK γ subunit as binding sites are confirmed. Meanwhile, the AMPK/unc-51-like autophagy-activating kinase 1 (ULK1)/microtubule-associated protein 1 light chain 3 beta (LC3B) and AMPK/mammalian target of rapamycin (mTOR) signaling pathways involved in anti-AD effects of GENI. The findings provide a new perspective on treating neurodegenerative diseases by activating AMPK for the energy metabolism disorder. Full article

In single-cell analysis, biological variability can be attributed to individual cells, their specific state, and the ability to respond to external stimuli, which are determined by protein abundance and their relative alterations. Mass spectrometry (MS)-based proteomics (e.g., SCoPE-MS and SCoPE2) can be used as a non-targeted method to detect molecules across hundreds of individual cells. To achieve high-throughput investigation, novel approaches in Single-Cell Proteomics (SCP) are needed to identify and quantify proteins as accurately as possible. Controlling sample preparation prior to LC-MS analysis is critical, as it influences sensitivity, robustness, and reproducibility. Several nanotechnological approaches have been developed for the removal of cellular debris, salts, and detergents, and to facilitate systematic sample processing at the nano- and microfluidic scale. In addition, nanotechnology has enabled high-throughput proteomics analysis, which have required the improvement of software tools, such as DART-ID or DO-MS, which are also fundamental for addressing key biological questions. Single-cell proteomics has many applications in nanomedicine and biomedical research, including advanced cancer immunotherapies or biomarker characterization, among others; and novel methods allow the quantification of more than a thousand proteins while analyzing hundreds of single cells. Full article

In current food safety monitoring, lateral flow immunoassays (LFIAs) are widely used for rapid food contaminant screening. Recent advances include smartphone readouts, offering semi-quantitative analysis of LFIAs with time, location, and data transfer in case of on-site testing. Following the screening, the next step in the EU regulations is confirmation by, e.g., liquid chromatography-tandem mass spectrometry (LC-MS/MS). In this work, using direct analysis in real time ambient ionization and triple quadrupole MS/MS (DART-QqQ-MS/MS), we achieved rapid confirmation of the identity of the substance(s) causing the LFIA result. In the workflow proposed, an individual performs the (on-site) smartphone LFIA screening, and when the result is suspect, an identification LFIA (ID-LFIA) strip is developed with the same sample extract. The ID-LFIA can be dissociated and rapidly analyzed in a control laboratory with DART-QqQ-MS/MS. The ID-LFIA consists of multiple lines of monoclonal antibodies against the mycotoxin deoxynivalenol, acting as a bioaffinity trap. The ID-LFIA/DART-QqQ-MS/MS approach has been developed and validated, along with the screening smartphone LFIA, and has demonstrated its applicability by analyzing incurred and spiked samples. The developed approach has been critically compared with our previous direct electrospray ionization MS method and was found to provide highly complementary information on the total deoxynivalenol contamination in the sample. Full article

Erlotinib inhibits epithelial growth factor receptor (EGFR) kinase activity and is used to treat non-small cell lung cancer (NSCLC). Despite its high efficacy, recurrence can occur in patients who become resistant to the drug. To address the underlying mechanism of Erlotinib resistance, we investigated additional mechanisms related to mode-of-drug-action, by multiple protein-binding interactions, besides EGFR by using drug affinity responsive target stability (DARTS) and liquid chromatography-mass spectrometry (LC-MS/MS) methods with non-labeled Erlotinib. DNA polymerase alpha subunit B (POLA2) was identified as a new Erlotinib binding protein that was validated by the DARTS platform, complemented with cellular thermal shift assays. Genetic knock-down of POLA2 promoted the anti-proliferative effect of the drug in the Erlotinib-resistant cell line H1299 with high POLA2 expression, whereas the overexpression of POLA2 restored anti-proliferative effects in the Erlotinib-sensitive cell line HCC827 with low POLA2 expression. Importantly, POLA2 expression levels in four NSCLC cell lines were positively correlated with anti-proliferative Erlotinib efficacy (Pearson correlation coefficient, R = 0.9886). These results suggest that POLA2 is a novel complementary target protein of Erlotinib, and could clinically provide validity as a surrogate marker for drug resistance in patients with NSCLC. Full article

Manipulating autophagy is a promising strategy for treating cancer as several autophagy inhibitors are shown to induce autophagic cell death. One of these, autophagonizer (APZ), induces apoptosis-independent cell death by binding an unknown target via an unknown mechanism. To identify APZ targets, we used a label-free drug affinity responsive target stability (DARTS) approach with a liquid chromatography/tandem mass spectrometry (LC–MS/MS) readout. Of 35 protein interactors, we identified Hsp70 as a key target protein of unmodified APZ in autophagy. Either APZ treatment or Hsp70 inhibition attenuates integrity of lysosomes, which leads to autophagic cell death exhibiting an excellent synergism with a clinical drug, temozolomide, in vitro, in vivo, and orthotropic glioma xenograft model. These findings demonstrate the potential of APZ to induce autophagic cell death and its development to combinational chemotherapeutic agent for glioma treatment. Collectively, our study demonstrated that APZ, a new autophagy inhibitor, can be used as a potent antitumor drug candidate to get over unassailable glioma and revealed a novel function of Hsp70 in lysosomal integrity regulation of autophagy. Full article