CT Quantification of Interstitial Lung Abnormalities and Changes of Agerelated Pathomorphology.

BACKGROUND: Interstitial lung abnormalities (ILA) are associated with further disease progression, increased mortality risk, and decline in lung function in the elderly, which deserves enough attention. OBJECTIVE: The objective of this study was to quantify the extent of interstitial lung abnormalities (ILA) in a non-smoking asymptomatic urban cohort in China using low-dose CT (LDCT) and to analyze the age-related pathological changes. METHODS: We retrospectively analyzed clinical data and chest LDCT images from a cohort of 733 subjects who were categorized into 3 groups: 18-39, 40-59, and ≥60 years old according to age. Furthermore, we selected 40 cases of wax-embedded lung tissue blocks archived after pulmonary bullectomy and the same age groups were categorized. Four representative CT signs of ILA, including interlobular septal thickening (ILST), intralobular interstitial thickening (ILIT), ground-glass opacity (GGO), and reticular shadow (RS), were semi-quantified based on the percentage of the affected area. The scores and distribution of four CT signs of ILA were compared between different sex and age groups. The age-related pathological changes were analyzed. RESULTS: The ILA findings were found predominantly in the lower lobes and the subpleural region. The semi-quantitative scores of four CT signs in all subjects under 40 were 0. However, in subjects over 40 years old, the scores gradually increased with age, although most of them remained low. The size of the alveoli increased, the number of alveoli decreased, the alveolar septum became thinner, and the number of ATII cells increased with age. A statistically significant difference was observed among the different age groups (χ2=50.624, P=0.033; χ2=80.000, P=0.043; χ2=33.833, P=0.000; χ2=13.525, P=0.031). The macrophage population and the percentage of collagen fibers in the alveolar septum increased, while the percentage of elastic fibers decreased with age. There was no significant difference among the different age groups (χ2=19.817, P=0.506; χ2=52.419, P=0. 682; χ2=54.868, P=0.518). CONCLUSION: When the four CT signs mentioned above are in the upper central area, and the score has a medium or high score, it is crucial to determine the underlying pathological causes. ILA may be the result of chronic lung injury.

Addressing the untargeted lipidomics challenge in urine samples: Comparative study of extraction methods by UHPLC-ESI-QTOF-MS.

Urine analysis has remained a fundamental and widely used method in clinical diagnostics for over a century. With its minimal invasive nature and comprehensive range of analytes, urine has established itself as a clinical diagnostic tool for various disorders, including renal, urological, metabolic, and endocrine diseases. Furthermore, urine's unique attributes make it an attractive matrix for biomarker discovery, as well as in assessing the metabolic and physiological states of patients and healthy individuals alike. However, limitations in our knowledge of average values and sources of urinary lipids decrease the wider clinical application of urinary lipidomics. In this context, untargeted lipidomics analysis relies heavily on the extraction and analysis of lipids in biological samples. Nevertheless, this type of analysis presents challenges in lipid identification due to the diverse nature of lipids. Therefore, proper sample treatment before analysis is crucial to obtain robust and reproducible lipidomic profiles. To address this gap, we conducted a comparative study of a urine pool sample collected from twenty healthy volunteers using four different lipid extraction methods: one biphasic and three monophasic protocols. The extracted lipids were then analyzed using UHPLC-MS and MS/MS, and the semi-quantification of all the accurately annotated lipid species was performed for each extraction method.

Simplified drug efficacy evaluation system for vasopressin neurodegenerative disease using mouse disease-specific induced pluripotent stem cells.

Familial neurohypophyseal diabetes insipidus (FNDI) is a degenerative disorder in which vasopressin-secreting neurons degenerate over time due to the production of mutant proteins. We have demonstrated therapeutic effects of chemical chaperones in an FNDI mouse model, but the complexity and length of this evaluation were problematic. In this study, we established disease-specific mouse induced pluripotent stem cells (iPSCs) from FNDI-model mice and differentiated vasopressin neurons that produced mutant proteins. Fluorescence immunostaining showed that chemical chaperones appeared to protect vasopressin neurons generated from iPSCs derived from FNDI-model mice. Although KCL stimulation released vasopressin hormone from vasopressin neurons generated from FNDI-derived iPSCs, vasopressin hormone levels did not differ significantly between baseline and chaperone-added culture. Semi-quantification of vasopressin carrier protein and mutant protein volumes in vasopressin neurons confirmed that chaperones exerted a therapeutic effect. This research provides fundamental technology for creating in vitro disease models using human iPSCs and can be applied to therapeutic evaluation of various degenerative diseases that produce abnormal proteins.

HRMS-based suspect screening of pharmaceuticals and their transformation products in multiple environmental compartments: An alternative to target analysis?

The comprehensive monitoring of pharmaceutically active compounds (PhACs) in the environment is challenging given the myriad of substances continuously discharged, the increasing number of new compounds being produced (and released), or the variety of the associated human metabolites and transformation products (TPs). Approaches such as high-resolution mass spectrometry (HRMS)-based suspect analysis have emerged to overcome the drawbacks of classical target analytical methods, e.g., restricted chemical coverage. In this study, we assess the readiness of HRMS-based suspect screening to replace or rather complement target methodologies by comparing the performance of both approaches in terms of i) detection of PhACs in various environmental samples (water, sediments, biofilm, fish plasma, muscle and liver) in a field study; ii) PhACs (semi)quantification and iii) prediction of their environmental risks. Our findings revealed that target strategies alone significantly underestimate the variety of PhACs potentially impacting the environment. However, relying solely on suspect strategies can misjudge the presence and risk of low-level but potentially risky PhACs. Additionally, semiquantitative approaches, despite slightly overestimating concentrations, can provide a realistic overview of PhACs concentrations. Hence, it is recommended to adopt a combined strategy that first evaluates suspected threats and subsequently includes the relevant ones in the established target methodologies.

A rapid and high-throughput multiplex genetic detection assay for detection, semi-quantification and virulence genotyping of Helicobacter pylori in non-invasive oral samples.

Aim: This study established a high-throughput multiplex genetic detection assay (HMGA) for rapid identification, semi-quantification and virulence analysis of Helicobacter pylori directly from the clinical non-invasive oral samples. Methods: The gastric mucosa and oral samples were collected from 242 patients in Shanghai from 2021 to 2022. All the samples were detected by routine clinical tests for H. pylori and Sanger sequenced for inconsistent results. A new multiplex PCR assay providing results within 4 hours was designed and optimized involving fluorescent dye-labeled specific primers targeted 16S rRNA gene, semi-quantitative gene ureC and 10 virulence genes of H. pylori. Semi-quantification was carried out by simulating the serial 10-fold dilutions of positive oral samples, and the H. pylori loads in different clinical samples were further compared. The mixed plasmids of virulence genes vacA s1, vacA m1 and vacA m2 were used to evaluate the performance on different genotypes. The consistency of 10 virulence genes in gastric mucosa, saliva, mouthwash and dental plaque of H. pylori-positive patients was compared. Results: The non-invasive HMGA was highly specific for detection of all 12 targets of H. pylori and human internal reference gene ß-globin, and the sensitivity to all target genes could reach 10 copies/µL. Compared with routine clinical tests and sequencing, non-invasive HMGA has a high level (>0.98) of sensitivity, specificity, accuracy, PPV, NPV and kappa coefficient for direct detection of H. pylori in oral samples. Moreover, by detecting peak area levels of ureC, it was confirmed that the H. pylori loads in gastric mucosa were significantly higher than those of the three kinds of oral samples (p<0.05). We also found that 45.0% (91/202) of patients had different H. pylori virulence genes in different oral samples. The concordance of positive detection rates of each virulence gene between saliva and gastric mucosa was more than 78% (p<0.05). Conclusion: The non-invasive HMGA proved to be a reliable method for the rapid H. pylori identification, semi-quantification and detection of 10 virulence genes directly in oral samples, providing a new idea for non-invasive detection of H. pylori.

Optimisation and implementation of QuEChERS-based sample preparation for identification and semi-quantification of 694 targeted contaminants in honey, jam, jelly, and syrup by UHPLC-Q/ToF high-resolution mass spectrometry.

A screening and semi-quantitative method was developed for the analysis of 694 various contaminants in honey, jam, jelly and syrup samples by ultrahigh-performance liquid chromatography and quadrupole time-of-flight mass spectrometry. Sample preparation, which was optimised using split factorial design, was based on acetate-buffered version of QuEChERS, followed by a clean-up step and a concentration step to enhance sensitivity of analytes. The method was validated according to SANTE/11312/2021 guidelines. The screening detection and limits of identification were established as being less than or equal to 0.05 mg.kg-1 for 89% and 74% of the contaminants, respectively. The validated screening method was applied to 50 concentrated sugary products. Overall, 46% of the samples were positive to pesticide residues. Most of the positive samples (78%) contained mixtures of pesticide residues. Three time-and-cost saving convenient strategies suitable for high-throughput analysis were proposed for the targeted semi-quantification of the previously contaminants identified in samples.

Revisit of the CatLet (Hexu) angiographic scoring system: a narrative review.

Background and Objective: We have recently developed the Coronary Artery Tree description and Lesion EvaluaTion (CatLet or Hexu, invented by He and Xu) angiographic scoring system, which, considering the coronary anatomy in its diversity, the stenosis degree of a coronary artery, and the myocardial territory subtended by the diseased coronary artery, can be utilized to predict clinical outcomes for patients with acute myocardial infarction (available at www.catletscore.com). Its values for clinical practice and coronary artery disease research are building upon. Over the past two years, the principles underlying this novel angiographic scoring system do not materially change although slight adjustments have really happened. Given these adjustments and the scoring experience gained in daily use, we think that it is necessary to elaborate on these points so that readers with interest are capable to better use this CatLet or Hexu angiographic scoring system both in clinical practice and in scientific research. Methods: The principles underlying this novel angiographic scoring system include the 17-myocardial segmental model, law of competitive blood supply, and law of flow conservation. Key Content and Findings: The adjustments made to this novel angiographic scoring system include: (I) the short axis of the left ventricle at the basal level is used to characterize the six types of right coronary artery size; (II) segments marked with 'X and 'S have a unified preset difference of one segment as adopted in the characterization of left anterior descending artery; (III) segments marked with '+ have been added to explain the rare variability in the obtuse marginal branches or in the posterolateral vessels in some cases. The CatLet or Hexu angiographic scoring system strictly follows the law of flow conservation in weighting assignment, and the lesion scoring correction has been additionally emphasized and detailed. Conclusions: The elaboration on these adjustments and scoring experience gained on the CatLet or Hexu angiographic scoring system will help to boost its use in cardiovascular field. The utilities of this novel angiographic scoring system have been preliminarily validated and its future is deserving of being anticipated.

Identification and characterization of quaternary ammonium compounds in Flemish indoor dust by ion-mobility high-resolution mass spectrometry.

Quaternary ammonium compounds (QACs) are a class of surfactants commonly used in disinfecting and cleaning products. Their use has substantially increased during the COVID-19 pandemic leading to increasing human exposure. QACs have been associated with hypersensitivity reactions and an increased risk of asthma. This study introduces the first identification, characterization and semi-quantification of QACs in European indoor dust using ion-mobility high-resolution mass spectrometry (IM-HRMS), including the acquisition of collision cross section values (DTCCSN2) for targeted and suspect QACs. A total of 46 indoor dust samples collected in Belgium were analyzed using target and suspect screening. Targeted QACs (n = 21) were detected with detection frequencies ranging between 4.2 and 100 %, while 15 QACs showed detection frequencies > 90 %. Semi-quantified concentrations of individual QACs showed a maximum of 32.23 µg/g with a median ∑QAC concentration of 13.05 µg/g and allowed the calculation of Estimated Daily Intakes for adults and toddlers. Most abundant QACs matched the patterns reported in indoor dust collected in the United States. Suspect screening allowed the identification of 17 additional QACs. A dialkyl dimethyl ammonium compound with mixed chain lengths (C16:C18) was characterized as a major QAC homologue with a maximum semi-quantified concentration of 24.90 µg/g. The high detection frequencies and structural variabilities observed call for more European studies on potential human exposure to these compounds. For all targeted QACs, drift tube IM-HRMS derived collision cross section values (DTCCSN2) are reported. Reference DTCCSN2 values allowed the characterization of CCS-m/z trendlines for each of the targeted QAC classes. Experimental CCS-m/z ratios of suspect QACs were compared with the CCS-m/z trendlines. The alignment between the two datasets served as an additional confirmation of the assigned suspect QACs. The use of the 4bit multiplexing acquisition mode with consecutive high-resolution demultiplexing confirmed the presence of isomers for two of the suspect QACs.

Towards an Automated Approach to the Semi-Quantification of [18F]F-DOPA PET in Pediatric-Type Diffuse Gliomas.

BACKGROUND: This study aims to evaluate the use of a computer-aided, semi-quantification approach to [18F]F-DOPA positron emission tomography (PET) in pediatric-type diffuse gliomas (PDGs) to calculate the tumor-to-background ratio. METHODS: A total of 18 pediatric patients with PDGs underwent magnetic resonance imaging and [18F]F-DOPA PET, which were analyzed using both manual and automated procedures. The former provided a tumor-to-normal-tissue ratio (TN) and tumor-to-striatal-tissue ratio (TS), while the latter provided analogous scores (tn, ts). We tested the correlation, consistency, and ability to stratify grading and survival between these methods. RESULTS: High Pearson correlation coefficients resulted between the ratios calculated with the two approaches: ρ = 0.93 (p < 10-4) and ρ = 0.814 (p < 10-4). The analysis of the residuals suggested that tn and ts were more consistent than TN and TS. Similarly to TN and TS, the automatically computed scores showed significant differences between low- and high-grade gliomas (p ≤ 10-4, t-test) and the overall survival was significantly shorter in patients with higher values when compared to those with lower ones (p < 10-3, log-rank test). CONCLUSIONS: This study suggested that the proposed computer-aided approach could yield similar results to the manual procedure in terms of diagnostic and prognostic information.

Separation of flavonoid isomers by cyclic ion mobility mass spectrometry.

Analytical techniques, such as liquid chromatography coupled to mass spectrometry (LC-MS) or nuclear magnetic resonance (NMR), are widely used for characterization of complex mixtures of (isomeric) proteins, carbohydrates, lipids, and phytochemicals in food. Food can contain isomers that are challenging to separate, but can possess different reactivity and bioactivity. Catechins are the main phenolic compounds in tea; they can be present as various stereoisomers, which differ in their chemical properties. Currently, there is a lack of fast and direct methods to monitor interconversion and individual reactivity of these epimers (e.g. epicatechin (EC) and catechin (C)). In this study, cyclic ion mobility mass spectrometry (cIMS-MS) was explored as a potential tool for the separation of catechin epimers. Formation of sodium and lithium adducts enhanced IMS separation of catechin epimers, compared to deprotonation and protonation. Baseline separation of the sodium adducts of catechin epimers was achieved. Moreover, we developed a fast method for the identification and semi-quantification of cIMS-MS separated catechin epimers. With this method, it is possible to semi-quantify the ratio between EC and C (1:5 to 5:1, within 50-1200 ng mL-1) in food samples, such as tea. Finally, the newly developed approach for cIMS-MS separation of flavonoids was demonstrated to be successful in separation of two sets of positional isomers (i.e. morin, tricetin, and quercetin; and kaempferol, fisetin, luteolin, and scutellarein). To conclude, we showed that both epimers and positional isomers of flavonoids can be separated using cIMS-MS, and established the potential of this method for challenging flavonoid separations.

KASP genotyping and semi-quantitation of G275E mutation in the α6 subunit of Thrips palmi nAChR gene conferring spinetoram resistance.

BACKGROUND: Pesticide resistance is a long-standing and growing problem in the chemical control of invertebrate pests. Molecular diagnostic methods can facilitate pesticide resistance management by accurately and efficiently detecting resistant mutations and their frequency. In this study, the kompetitive allele specific PCR (KASP) approach, a technology for high-throughput single nucleotide polymorphism (SNP) genotyping, is validated as a useful method for characterizing genotypes at a pesticide-resistance locus for the first time. We focus on the spinetoram resistance mutation of G275E in the nicotinic acetylcholine receptor alpha 6 (nAChR α6) subunit gene of Thrips palmi. RESULTS: Of the 341 individuals of Thrips palmi tested, 98.24% were successfully genotyped, with 100% concordance with Sanger sequencing results. We then quantitatively mixed genomic DNA of known genotypes to establish 21 DNA mixtures with a resistant allele frequency ranging from 0 to 100% at steps of 5%. The linear discriminant analysis (LDA) showed that 75.8% of original grouped cases were correctly classified; six groups had no overlap in membership (resistant allele frequency: 0%, 5%, 10-75%, 80-85%, 90-95%, and 100%). When we chose 11 pooled samples with 10% steps for LDA, 84.4% of original grouped cases were correctly classified; seven groups had no overlap in membership (0%, 10%, 20-30%, 40-70%, 80%, 90%, 100%). The results indicated that KASP applied to pooled samples may provide a semi-quantitative estimate of resistance. CONCLUSIONS: Our study points to the suitability of KASP for high-throughput genotyping of genotypes affecting pesticide resistance and semi-quantitative assessments of resistance allele frequencies in populations. © 2023 Society of Chemical Industry.

Development of semi-quantitative urinary sodium test strip.

Excessive salt consumption has been associated with greater risk of hypertension. Therefore, monitoring of dietary sodium consumption should be prioritized. As sodium is mainly excreted through urine, 24-h urine sample can be used to estimate individual sodium intake. Thus, a simple and inexpensive semi-quantitative urinary sodium detection test strip was developed based on the enzymatic reaction between ß-galactosidase and chlorophenol red-ß-d-galactopyranoside. When tested, color formation was distinguished at 0 M (chartreuse yellow), 0.05 M (sunflower), 0.1-0.15 M (mango tango), and 0.2-0.25 M (persimmon) sodium. Analysis from ImageJ showed a linear result (r2  > 0.9), low SD, and significant increase in magenta difference (p < 0.01) between 0 and 0.05-0.25 M sodium. Test strip can detect 0.03 M sodium at minimum but did not last for >2 days in adverse storage conditions (laboratory conditions, ∼80% relative humidity, 40°C, and direct light exposure) when stored in test strip bottles, and even shorter when exposed to the environment. The presence of urinary potassium, urea, and glucose did not affect test strip performance. Test strip produced comparable results to flame photometry with <15% variation when tested on overnight, random spot, and 24-h urine samples. Overall, the developed test strip can be used to enzymatically semi-quantify 0.05-0.25 M sodium.

[Occupational risk assessment levels before and after improvement on fueling devices in ammoniation facilities from 2 nuclear plants].

OBJECTIVE: To evaluate and compare possible changes of individual health risk levels for ammonia exposed positions from ammoniation facilities in 2 nuclear power plants(X and Y) through occupational risk assessment models(OHRA) before and after devices improvement. METHODS: This study tried to understand the improvement of fueling devices in ammoniation facilities, to identify the ammonia exposed positions and their working content, to collect information on protective measures and personal protective equipment situation through on-sites survey and detailed records of work days. Next, detection on C_(STE) of ammonia at different worksites and C_(TWA) of 10 involved positions were conducted, fresh air requirement and ventilation air changing rates were calculated through wind speed detection or collected directly. Finally, a semi-quantitative comprehensive index model and a quantitative non-carcinogenic model from standard GBZ/T 298-2017 were utilized to assess risk levels before and after improvement. RESULTS: After devices improvement, a series of innovations were observed, followed by:(1) ammonia steal tanks in stacking storage were replaced by metal pressure sealing tanks for reduction of exposure frequency and long time storage. (2) manual filling pattern was totally substituted by automatic filling devices with new installed ventilation systems and alarming apparatus. (3) increasing ventilation effect by doors opening when ammonia was filling. In this regard, conentration levels(C_(STE)) referred to positions of chemical analysis engineers, chemical sampling engineers, nuclear maintenance workers at 3# unit from X, and regular island operators at 1#unit, patrolling operators at 2# unit from Y were declined to qualified levels after improvement from unqualified levels before that, and significant difference of C_(STE )detected before and after improvement were found statistically(Z=10.856, P<0.001). C_(TWA )were all within the qualified ranges before and after improvement for relatively short cumulative exposure time. Moreover, the ventilation air changing rates from related indoor sectors were increased to 13.0 to 30.9 times/h after improvement which indicated a statistical difference as compared to quantitative ranges before started(Z=11.670, P<0.001). Further, a relative negative correlation was observed between C_(STE )and ventilation air changing rates(r=-0.39, P<0.05) while no correlation between them was spotted after improvement(r=-0.051, P>0.05). In addition, most positions like chemical analysis engineers and others changed their risk levels to a lower one from that they used to be after improvement with an observed significant difference(Z=1.345, P<0.05), by contrast, risk levels of positions like nuclear maintenance workers at 3#unit or patrolling operators at 4#unit increased a level or remained the same before and after improvement might be result ed from enhancement of cumulative exposure time. No significant difference among position risk levels under the quantitative non-carcinogenic model was observed(Z=0.447, P>0.05). CONCLUSION: The measure of devices improvement indeed decreased the ambient ammonia concentrations at workplaces of ammoniation facilities in X and Y and changed individual risk levels for most involved positions. The semi-quantitative comprehensive index model was more appropriated for self-contrast risk assessment application than quantitative non-carcinogenic model did, especially when improvement occurred.

Phosphoproteome profiling of hippocampal synaptic plasticity.

The balance between the actions of protein kinases and phosphatases is crucial for neuronal functions, including synaptic plasticity. Although the phosphorylation and dephosphorylation of neuronal proteins are regulated by synaptic plasticity, no systematic analyses of this have yet been conducted. We performed a phosphoproteomic analysis of hippocampal synaptic plasticity using a nano-Acquity/Synapt LC-MS/MS system. Neuronal proteins were extracted from hippocampal tissues and cultured neurons exposed to long-term potentiation (LTP) or long-term depression (LTD). Filter-aided sample preparation (FASP) was performed to remove residual anionic detergents for complete tryptic digestion. Phosphopeptides were then enriched using TiO2 chromatography, followed by immunoaffinity chromatography with an anti-phosphotyrosine antibody. Among the 1500 phosphopeptides identified by LC-MS/MS, 374 phosphopeptides were detected simultaneously in both hippocampal tissues and cultured neurons. Semi-quantification counting the number of spectra of each phosphopeptide showed that 42 of 374 phosphopeptides changed significantly depending on synaptic plasticity. In conclusion, a new proteomic method using sequential enrichment of phosphopeptides and semi-quantification enabled the phosphoproteomic analysis of hippocampal synaptic plasticity.

Identification and semi-quantification of metabolites of new plasticizers in urine collected from flemish adults and children.

A suspect screening workflow combined with a semi-quantification method was applied for the investigation of metabolites of the plasticizers di-propylene glycol dibenzoate (DiPGDB) and tri-n-butyl trimellitate (TBTM) in human urine collected from adults and children during winter (W) and summer (S) seasons. Liquid chromatography - quadrupole time of flight mass spectrometry (LC-QTOF-MS) was applied for the analyses. Two direct and one indirect metabolites of DiPGDB were identified: 3-(3-hydroxypropoxy) propyl benzoate (DiPGDB-M194), 3,4,5-trihydroxy-6-[3-(3-hydroxypropoxy) propoxy] oxane-2-carboxylic acid (DiPGDB-M310), hippuric acid (DiPGDB-M179) and one metabolite of TBTM: bis(butoxycarbonyl) benzoyloxy]-3,4,5-trihydroxyoxane-2-carboxylic acid (TBTM-M498). The identified metabolites were reported with levels of confidence (LoC) 2 and 3 and their concentrations were assessed using a semi-quantification approach. The respective concentration ranges for W and S samples were 0.20-42 ng/mL and 0.07-29 ng/mL for DiPGDB-M194, 2.5-1420 ng/mL and 5.0-2320 ng/mL for DiPGDB-M310, 230-10840 ng/mL and 320-8420 ng/mL for DiPGDB-M179, and 0.40-30 ng/mL and 0.65-30 ng/mL for TBTM-M498. The detection frequency order in urine samples was DiPGDB-M310 = DiPGDB-M179 (100%) >TBTM-M498 (44%) > DiPGDB-M194 (28%) for W and DiPGDB-M179 (99%)> DiPGDB-M310 (98%) > TBTM-M498 (57%) > DiPGDB-M194 (30%) for S. The identified metabolites DiPGDB-M310, DiPGDB-M194 and TBTM-M498 are potential biomarkers for the evaluation of human exposure to DiPGDB and TBTM. DiPGDB-M179 cannot be used for the same purpose due to its formation from compounds with multi-source origin. The application of the semi-quantification method could be useful for further studies where analytical standards are not available.

Qualitative and semi-quantitative screening of selected psychoactive drugs in blood: Usefulness of liquid chromatography - triple quadrupole and quadrupole time-of-flight mass spectrometry in routine toxicological analyses.

Routine toxicological analysis requires broad screening for a large number of therapeutically prescribed and other compounds, and/or their metabolites. This article specifically focuses on three classes of psychoactive substances: antidepressants (ADs), antipsychotics (APs) and benzodiazepines and Z-drugs (BZDs). Two screening methods were compared for their ease-of-use in a routine setting, based upon the analysis of 105 medico-legal case samples. Analytes of interest were extracted using liquid-liquid extraction and separated using liquid chromatography with a total run time of 12 min per sample. A first detection method used targeted triple quadrupole mass spectrometry, operated in triggered multiple reaction monitoring mode (tMRM). False negative results were noted for 15% of the total number of detected analytes only, the majority of which were either present at sub- to low therapeutic levels or were metabolites of other analytes in the samples. The occurrence of false positive results was rare. A second screening method used quadrupole time-of-flight mass spectrometry (QTOF) for untargeted data acquisition. Data analysis was facilitated by the creation of an in-house, subset mass spectral database. As was seen for the tMRM screening, false negative results were observed in less than 20% of the total number of detected analytes, either for compounds at low concentrations or of which metabolites could be identified in the samples. More false positive results were observed due to an observed bias for prothipendyl. Determination of the exact concentration in a sample may only be required depending on the specific case circumstances. For this purpose, semi-quantification using each of the screening methods was investigated. Excellent results were observed using the tMRM method in combination with a small number of labelled internal standards (n = 12). Semi-quantification using the QTOF screening method was more laborious, but limited results on selected compounds indicated equally good results. Overall, the developed semi-quantitative screening methods performed well and - following further validation on case samples - could be implemented for most compounds in routine toxicological analysis without the need for highly trained or specialised personnel.

Impact of roasting on the phenolic and volatile compounds in coffee beans.

Phenolic compounds present in coffee beans could generate flavor and bring benefits to health. This study aimed to evaluate the impacts of commercial roasting levels (light, medium, and dark) on phenolic content and antioxidant potential of Arabica coffee beans (Coffea arabica) comprehensively via antioxidant assays. The phenolic compounds in roasted samples were characterized via liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (LC-ESI-QTOF-MS/MS). Furthermore, the coffee volatile compounds were identified and semi-quantified by headspace/gas chromatography-mass spectrometry (HS-SPME-GC-MS). Generally, for phenolic and antioxidant potential estimation, light roasted samples exhibited the highest TPC (free: 23.97 ± 0.60 mg GAE/g; bound: 19.32 ± 1.29 mg GAE/g), DPPH, and FRAP. The medium roasted beans performed the second high in all assays but the highest ABTS+ radicals scavenging capacity (free: 102.37 ± 8.10 mg TE/g; bound: 69.51 ± 4.20 mg TE/g). Totally, 23 phenolic compounds were tentatively characterized through LC-ESI-QTOF-MS/MS, which is mainly adopted by 15 phenolic acid and 5 other polyphenols. The majority of phenolic compounds were detected in the medium roasted samples, followed by the light. Regarding GC-MS, a total of 20 volatile compounds were identified and semi-quantified which exhibited the highest in the dark followed by the medium. Overall, this study confirmed that phenolic compounds in coffee beans would be reduced with intensive roasting, whereas their antioxidant capacity could be maintained or improved. Commercial medium roasted coffee beans exhibit relatively better nutritional value and organoleptic properties. Our results could narrow down previous conflicts and be practical evidence for coffee manufacturing in food industries.

Use of Passive and Grab Sampling and High-Resolution Mass Spectrometry for Non-Targeted Analysis of Emerging Contaminants and Their Semi-Quantification in Water.

Different groups of organic micropollutants including pharmaceuticals and pesticides have emerged in the environment in the last years, resulting in a rise in environmental and human health risks. In order to face up and evaluate these risks, there is an increasing need to assess their occurrence in the environment. Therefore, many studies in the past couple of decades were focused on the improvements in organic micropollutants' extraction efficiency from the different environmental matrices, as well as their mass spectrometry detection parameters and acquisition modes. This paper presents different sampling methodologies and high-resolution mass spectrometry-based non-target screening workflows for the identification of pharmaceuticals, pesticides, and their transformation products in different kinds of water (domestic wastewater and river water). Identification confidence was increased including retention time prediction in the workflow. The applied methodology, using a passive sampling technique, allowed for the identification of 85 and 47 contaminants in the wastewater effluent and river water, respectively. Finally, contaminants' prioritization was performed through semi-quantification in grab samples as a fundamental step for monitoring schemes.

A novel workflow for semi-quantification of emerging contaminants in environmental samples analyzed by LC-HRMS.

There is an increasing need for developing a strategy to quantify the newly identified substances in environmental samples, where there are not always reference standards available. The semi-quantitative analysis can assist risk assessment of chemicals and their environmental fate. In this study, a rigorously tested and system-independent semi-quantification workflow is proposed based on ionization efficiency measurement of emerging contaminants analyzed in liquid chromatography-high-resolution mass spectrometry. The quantitative structure-property relationship (QSPR)-based model was built to predict the ionization efficiency of unknown compounds which can be later used for their semi-quantification. The proposed semi-quantification method was applied and tested in real environmental seawater samples. All semi-quantification-related calculations can be performed online and free of access at http://trams.chem.uoa.gr/semiquantification/ .

Spotlight on mass spectrometric non-target screening analysis: Advanced data processing methods recently communicated for extracting, prioritizing and quantifying features.

Non-target screening of trace organic compounds complements routine monitoring of water bodies. So-called features need to be extracted from the raw data that preferably represent a chemical compound. Relevant features need to be prioritized and further be interpreted, for instance by identifying them. Finally, quantitative data is required to assess the risks of a detected compound. This review presents recent and noteworthy contributions to the processing of non-target screening (NTS) data, prioritization of features as well as (semi-) quantitative methods that do not require analytical standards. The focus lies on environmental water samples measured by liquid chromatography, electrospray ionization and high-resolution mass spectrometry. Examples for fully-integrated data processing workflows are given with options for parameter optimization and choosing between different feature extraction algorithms to increase feature coverage. The regions of interest-multivariate curve resolution method is reviewed which combines a data compression alternative with chemometric feature extraction. Furthermore, prioritization strategies based on a confined chemical space for annotation, guidance by targeted analysis and signal intensity are presented. Exploiting the retention time (RT) as diagnostic evidence for NTS investigations is highlighted by discussing RT indexing and prediction using quantitative structure-retention relationship models. Finally, a seminal technology for quantitative NTS is discussed without the need for analytical standards based on predicting ionization efficiencies.