Liquid Chromatography Quadrupole Time-of-flight Mass Spectrometry Research Articles

Semi refined carrageenan-nanocomposite film incorporated with Bentong ginger extracts for active food packaging: Synthesis and characterization

Bentong Ginger (BG), Malaysia patented ginger consist of various bioactive compounds such as gingerols which account for the various pharmacological benefits in human including antioxidant, antimicrobial, and anticancer. The objective of this work was to analyze the effectiveness of BG extracts as an antioxidant additive in plasticized semi refined carrageenan-nanocellulose (CN) to produce active packaging films at concentrations ranging from 5 to 20 % (v/v). The bioactive constituents in BG extracts were discovered using Liquid Chromatography Quadrupole Time-Of-Flight Mass Spectrometry (LC-QTOF/MS) and the possible interaction between the semi refined carrageenan-nanocellulose film network and BG were revealed using Fourier-Transform Infrared (FTIR) Spectroscopy. The effects of BG extracts formulated in CN film on the antioxidant release, physical and mechanical properties of the films were studied. The CN film with 20% BG improved the most, exhibiting excellent mechanical and physical properties as well as antioxidant release tests that revealed the highest release of antioxidant and total phenolic content in 95 % food simulants up to 28 days storages. This study validated that incorporation of BG extract can a promising natural additive for active packaging materials.

Comprehensive testing and rapid dissemination of local drug supply surveillance data in Rhode Island.

The North American overdose crisis has continued at unprecedented rates with more than 100,000 overdose deaths estimated to have occurred in the United States in 2022. Regional variations in overdose rates signify differences in local drug supplies. State-level drug supply surveillance systems have been limited in their ability to document and communicate the rapidly changing drug supplies which can hinder harm reduction efforts at the community level. We sought to address by piloting a two-year, community-engaged local drug supply surveillance program in Rhode Island (RI). The first set of samples (n=125) were collected from May 2022 to January 2023 across RI and included used paraphernalia (e.g., cookers), refuse (e.g., baggies), and product. Samples were tested using comprehensive toxicology testing approaches via liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). Results were disseminated to participants and the broader public across platforms. Fentanyl was detected in 67.2% of all samples tested. 39.2% (n=49) of samples were expected to be fentanyl. Xylazine was detected in 41.6% of all samples-always in combination with fentanyl-and no samples were expected to contain xylazine. In expected stimulant samples (n=39), 10% contained fentanyl and/or analogues as major substances and 30.8% contained trace amounts of fentanyl and/or analogues. In expected stimulant samples, 15.4% contained xylazine with fentanyl. No opioids or benzodiazepines were detected in expected hallucinogen or dissociative samples (n=7). In expected benzodiazepine samples (n=8), no opioids were detected. Our results describe part of the local drug supply in Rhode Island, including a presence of NPS and adulterants (e.g., designer benzodiazepines, xylazine). Importantly, our findings underscore the feasibility of developing a community-driven drug supply surveillance database. Expanding drug supply surveillance initiatives is imperative for improving the health and safety of people who use drugs and informing public health approaches to addressing the overdose crisis.

Wound Healing Effect of Supercritical Carbon Dioxide Datura metel L. Leaves Extracts: An In Vitro Study of Anti-Inflammation, Cell Migration, MMP-2 Inhibition, and the Modulation of the Sonic Hedgehog Pathway in Human Fibroblasts.

Datura metel L. (thorn apple) has been used in Thai folk wisdom for wound care. In this study, we chose supercritical carbon dioxide extraction (scCO2) to develop crude extraction from the leaves of the thorn apple. The phytochemical profiles were observed using liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS). The biological activities of D. metel were performed through antioxidant assays, anti-inflammation based on the Griess reaction, the migration assay, the expression of matrix metalloproteinase-2 (MMP-2), and regulatory genes in fibroblasts. Dm1 and Dm2 extracts were obtained from scCO2 procedures at different pressures of 300 and 500 bar, respectively. Bioactive compounds, including farnesyl acetone, schisanhenol B, and loliolide, were identified in both extracts. The antioxidant properties of both D. metel extracts were comparable to those of l-ascorbic acid in hydrogen peroxide-induced fibroblasts with no significant difference. Additionally, Dm1 and Dm2 significantly inhibited the nitrite production levels of 1.23 ± 0.19 and 1.52 ± 0.05 μM, respectively, against the lipopolysaccharide-treated group (3.82 ± 0.39 μM). Interestingly, Dm1 obviously demonstrated the percentage of wound closure with 58.46 ± 7.61 and 82.62 ± 6.66% after 36 and 48 h of treatment, which were comparable to the commercial deproteinized dialysate from the calf blood extract. Moreover, both extracts were comparable to l-ascorbic acid treatment in their ability to suppress the expression of MMP-2: an enzyme that breaks down collagen. The gene expressions of SHH, SMO, and GLI1 that control the sonic hedgehog pathway were also clearly upregulated by Dm1. Consequently, the scCO2 technique could be applied in D. metel extraction and contribute to potentially effective wound closure.

Simultaneous determination of 11 nootropic substances potentially adulterated in dietary supplements for improving brain function using ultra-performance liquid chromatography and liquid chromatography–quadrupole-time-of-flight mass spectrometry

Recently, the demand for improved brain function and concentration has increased in the dietary supplement market. However, to artificially enhance their pharmacological efficacy, dietary supplements may be illegally adulterated with unauthorised substances. Therefore, we developed a rapid and accurate method to simultaneously determine 11 nootropic substances using an ultra-high-performance liquid chromatography (UPLC) system equipped with a photodiode array (PDA) detector. In addition, sample preparation procedures were semi-optimised for various types of matrices, including solid (hard capsule, tablet, powder, and pill) and liquid (oil and extract) samples. The method was validated to determine the limit of detection (LOD), limit of quantification (LOQ), method detection limit (MDL), method quantitation limit (MQL), specificity, linearity, precision, accuracy, recovery, stability, and matrix effects. The validation results satisfied international validation guideline requirements. To test the applicability of the method, 55 real samples advertised as effective brain health, memory, and cognition supplements were analysed. Among the real samples, vinpocetine (2.483 and 7.296 µg/g), and kavain (69–44.056 µg/g) were detected. In addition, the detected compounds were confirmed by comparing their fragmentation patterns with those of the reference standards using liquid chromatography–quadrupole-time-of-flight mass spectrometry (LC-QTOF/MS). In conclusion, the UPLC-PDA method not only rapidly and accurately quantifies illegal nootropics but also enables the pre-emptive investigation and identification of 11 nootropic substances in illegal dietary supplements to protect public health.

Frequency of new psychoactive substances in hair and urine samples of individuals subject to drug testing in driving license regranting-A toxicological perspective.

In recent years, numerous new psychoactive substances (NPS) have emerged on the illicit drug market. The assumed non-detectability of these drugs is often a key motivation for individuals subject to drug testing, such as those in driving license regranting programs. In these programs, NPS are not routinely tested for, and thus, subjects who have to prove abstinence from common drugs of abuse might switch to NPS to avoid positive drug tests. The aim of the study was to determine the frequency of these substances in the hair and urine samples of individuals undergoing drug testing in driving license regranting. A total of 1037 samples (577 hair and 460 urine samples) collected from 949 subjects between February 2017 and December 2018 were retrospectively analyzed for designer drugs and synthetic cannabinoids by liquid chromatography-quadrupole-time-of-flight-mass spectrometry (LC-QTOF-MS). For a more sensitive analysis of synthetic cannabinoids and their metabolites, additional testing was performed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Overall, 42 hair and two urine samples, which were obtained from 40 subjects, tested positive for NPS resulting in a frequency of 4.2%. While synthetic cannabinoids were detected in all cases, designer drugs were only found in three of these cases. With regard to the 577 hair samples analyzed, 7.3% screened positive, whereas only 0.4% of the 460 tested urine samples contained NPS. The results of this study indicate that synthetic cannabinoid use seems to be popular among this population, and therefore, testing for synthetic cannabinoids should be requested more often preferably using hair analysis.

Anti-inflammatory effect and component analysis of Chaihu Qingwen granules

Ethnopharmacological relevanceAs prevalent acute respiratory condition in clinical practice, acute lung injury has a quick start and severe symptoms which can harm patients physically. Chaihu Qingwen granules (CHQW) is a classic formula for the treatment of respiratory diseases. Clinical observation shows that CHQW has good efficacy in treating colds, coughs, and fevers. Aim of the studyThe aim of this study was to investigate the anti-inflammatory effect of CHQW on lipopolysaccharide (LPS)-induced acute lung injury (ALI) model in rats and to explore its potential mechanism, as well as to clarify its substance composition. Materials and methodsMale SD rats were randomly divided into the blank group, the model group, the ibuprofen group, the Lianhua Qingwen capsule group and the CHQW group (2, 4 and 8 g/kg, respectively). The LPS-induced acute lung injury (ALI) model in rats was established after pre-administration. The histopathological changes in the lung and the levels of inflammatory factors in bronchoalveolar lavage fluid (BALF) and serum of ALI rats were observed. The inflammation-related proteins toll-like receptor 4 (TLR4), inhibitory kappa B alpha (IκBα), phospho-IκBα (p-IκBα), nuclear-factor-kappa B (NF-κB), and NLR family pyrin domain containing 3(NLRP3) expression levels were measured by western blotting analysis and immunohistochemical analysis. The chemical composition of CHQW was identified by liquid chromatography-quadrupole-time of flight-mass spectrometry (LC-Q-TOF-MS). ResultsCHQW significantly ameliorated lung tissue pathological injury in LPS-induced ALI rats and decreased the release of inflammatory cytokines (interleukin-1β, interleukin-17 and tumor necrosis factor-α) in BALF and serum. In addition, CHQW decreased the expression of TLR4, p-IκBα and NF-κB proteins, increased the level of IκBα, regulated the TLR4/NF-κB signaling pathway, and inhibited the activation of NLRP3. The chemical components of CHQW were analyzed by LC-Q-TOF-MS, and a total of 48 components were identified by combining information from the literature, mainly flavonoids, organic acids, lignans, iridoids and phenylethanoid glycosides. ConclusionThe results of this study showed that the pretreatment of CHQW had a strong protective effect on LPS-induced ALI in rats, reducing lung tissue lesions and decreasing inflammatory cytokines released in BALF and serum. The protective mechanism of CHQW may be related to the inhibition of the TLR4/NF-κB signaling pathway and NLRP3 activation. The main active ingredients of CHQW are flavonoids, organic acids, lignans, iridoids and phenylethanoid glycosides.

Classification of Angelica species found in various foods using an LC-QTOF/MS-based metabolomics approach

In Korea, Angelica gigas is commonly known as Danggui. However, two other species on the market, Angelica acutiloba and Angelica sinensis, are also commonly called Danggui. Since the three Angelica species have different biologically active components, thus, different pharmacological activities, clear discrimination between them is needed to prevent their misuse. A. gigas is used not only as a cut or powdered product but also in processed foods, where it is mixed with other ingredients. To discriminate between the three Angelica species, reference samples were analysed as non-targeted using liquid chromatography-quadrupole time of flight/mass spectrometry (LC-QTOF/MS) and a metabolomics approach in which a discrimination model was established by partial least squares-discriminant analysis (PLS-DA). Then, the Angelica species in the processed foods were identified. First, 32 peaks were selected as marker compounds and a discrimination model was created using PLS-DA, and its validation was confirmed. Classification of the Angelica species was undertaken using the YPredPS value, and it was confirmed that all 21 foods examined contained the appropriate Angelica species indicated on the product packaging. Likewise, it was confirmed that all three Angelica species were accurately classified in the samples to which they were added.

Magnetic graphitized carbon black based on crystal growth method combined with high-resolution mass spectrometry for screening of 300 pesticide residues in Radix Codonopsis and Angelica sinensis

In this study, a high-throughput method for analyzing 300 pesticide residues in Radix Codonopsis and Angelica sinensis was established by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) using iron tetroxide loaded graphitized carbon black magnetic nanomaterial (GCB/Fe3O4) as the purification material. It was optimized that saturated salt water and 1 % acetate acetonitrile were used as the extraction solution, then the supernatant was purified with 2 g anhydrous CaCl2 and 300 mg GCB/Fe3O4. As a result, 300 pesticides in Radix Codonopsis and 260 in Angelica sinensis achieved satisfactory results. The limits of quantification of 91 % and 84 % of the pesticides in Radix Codonopsis and Angelica sinensis reached 10 μg/kg, respectively. The matrix-matched standard curves ranging from 10 to 200 μg/kg were established with correlation coefficients (R) above 0.99. The pesticides meeting SANTE/12682/2021 accounted for 91.3 %, 98.3 %, 100.0 % and 83.8 %, 97.3, 100.0 % of the total pesticides added in Radix Codonopsis and Angelica sinensis respectively, which were spiked at 10, 20,100 μg/kg. The technique was applied to screen 20 batches of Radix Codonopsis and Angelica sinensis. Five pesticides were detected, three of which were prohibited according to the Chinese Pharmacopoeia (2020 Edition). The experimental results showed that GCB/Fe3O4 coupled with anhydrous CaCl2 exhibited good adsorption performance and could be used for sample pretreatment of various pesticide residues in Radix Codonopsis and Angelica sinensis. Compared with the reported methods for determining pesticides in traditional Chinese medicine (TCM), the proposed method has the advantage of less time-consuming in the clean-up procedure. Furthermore, as a case study on root TCM, this approach may serve as a reference for other TCM.

Analysis of 13 Banned Colorants in Cosmetics via Liquid Chromatographic and Mass Spectrometric Techniques

Even though synthetic colorants can cause side effects such as allergies and pigmentation, they have not been sufficiently researched. Herein, high-performance liquid chromatography, liquid chromatography-tandem mass spectrometry, and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) were used to detect 13 banned synthetic colorants in cosmetics and characterize their fragmentation. The developed HPLC method was validated following the International Conference on Harmonisation guidelines (specificity, limit of detection, limit of quantification, recovery, linearity, accuracy, and precision) and applied to 120 distributed cosmetic products, one of which was found to contain three illegal synthetic colorants, namely Basic Blue 26 (0.33 mg/g), Basic Red 2 (0.53 mg/g), and Basic Yellow 28 (31.50 mg/g). Additionally, based on their fragment ions obtained using LC-Q-TOF-MS, the fragmentation pattern of synthetic colorants was predicted. Thus, our work paves the way for the reliable detection of illegal synthetic colorants and may help to prevent the distribution of cosmetics containing the same.

Enhanced removal of Sb(III) and Sb(V) using biosynthesized iron sulfide nanoparticles: Identification of biomolecules in the capping layer and its functions

As a green and sustainable technology, biosynthesis of iron sulfide nanoparticles (FeS) has attracted increasing recent attention. However, a comprehensive analysis of the biomolecular structure and function of the capping layer is currently lacking and is now considered essential because the reactivity of biosynthesized FeS is believed to be highly dependent on this capping layer. Here, FeS (PML-FeS) synthesized by Pinus massoniana Lamb extracts were used to remove Sb(III) and Sb(V) and in so doing gain a better understanding of the function of the capping layer via comparison with the removal of both Sb species by bare-FeS. The maximum removal efficiencies of Sb(III) and Sb(V) (100 and 87.6%) obtained by PML-FeS, were much higher than that obtained by bare-FeS (78.7 and 61.2%), indicating that the biomolecules in the capping layer enhanced the removal capacity. Advanced characterizations confirmed that the removal of Sb(III) and Sb(V) by the capping layer was based predominantly on adsorption, while the removal of Sb(III) and Sb(V) by bare-FeS exhibited both adsorption and oxidation behavior. This suggested a synergistic effect in PML-FeS between the capping layer and bare-FeS. Liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) confirmed that biomolecules in the capping layer were enriched in –OH and –COOH, demonstrating that oxygen-containing functional groups play a significant role in provision of active binding sites for Sb removal.

The Differences of Nutrient Components in Edible and Feeding Coix Seed at Different Developmental Stages Based on a Combined Analysis of Metabolomics.

Coix lachryma-jobi L. is an excellent plant resource that has a concomitant function for medicine, foodstuff and forage in China. At present, the commonly used cultivar for both medicine and foodstuff is Xiaobaike, and the cultivar for foraging is Daheishan. However, differences in the internal composition of plants lead to the expression of different phenotypic traits. In order to comprehensively elucidate the differences in nutrient composition changes in Coix seeds, a non-targeted metabolomics method based on ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) was used to analyze the metabolic changes in Coix seeds at different developmental stages. An edible Coix relative (Xiaobaike) and a feeding Coix relative (Daheishan) were selected as the research subjects. In the metabolome analysis of Coix seed, 314 metabolites were identified and detected, among which organic acids, carbohydrates, lipids, nucleotides and flavonoids were the main components. As an important standard for evaluating the quality of Coix seed, seven lipids were detected, among which fatty acids included not only even-chain fatty acids, but also odd-chain fatty acids, which was the first time detecting a variety of odd-chain fatty acids in Coix seed. The analysis of the compound contents in edible and feeding-type Coix lachryma-jobi L. and the lipid content at the mature stage showed that, among them, arachidic acid, behenic acid, heptadecanoic acid, heneicosanoic acid and pristanic acid may be the key compounds affecting the lipid content. In addition, in the whole process of semen coicis maturation, edible and feeding Coix show similar trends, and changes in the third period show clear compounds in the opposite situation, suggesting that edible and feeding Coix not only guarantee the relative stability of species but also provide raw materials for genetic breeding. This study provides valuable information on the formation of the edible and medicinal qualities of Coix.

Determination of 11 Cannabinoids in Cannabis sativa L. by Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-Q-TOF-MS)

Eleven major cannabinoids from each subdivided tissue of drug-type and fiber-type cannabis plants were determined by means of a liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). The cannabinoids analyzed in this study were tetrahydrocannabinol acid (THCA), Δ9-tetrahydrocannabinol (Δ9-THC), cannabidiol acid (CBDA), cannabidiol (CBD), Δ8-tetrahydrocannabinol (Δ8-THC), cannabinol (CBN), cannabichromene (CBC), cannabidivarin (CBDV), cannabigerolic acid (CBGA), cannabigerol (CBG) and tetrahydrocannabivarin (THCV). As a result, THCA was detected in the bracts at 28.4 µg/mg, in the buds at 24.8 µg/mg, and in the leaves at 5.1 to 10.5 µg/mg in the drug-type cannabis plant. In addition, Δ9-THC, CBGA, CBN, CBG, CBC, and THCV were mainly detected in bracts, buds, and leaves. On the other hand, as for the fiber-type cannabis plant, CBDA was detected in the bracts at 27.5 µg/mg, in the buds at 10.6 µg/mg, and in the leaves at 1.5-3.3 µg/mg. In addition, Δ9-THCA, CBD, Δ9-THC, CBC, and CBG were mainly detected in bracts, buds, and leaves.

Multi-Dimensional Antioxidant Screening of Selected Australian Native Plants and Putative Annotation of Active Compounds.

Acacia implexa, Eucalyptus rossii and Exocarpos cupressiformis are native plants of Australia, which were used by the First Peoples for medicinal purposes. In this study, 70% aqueous ethanol crude extracts were prepared from A. implexa bark and leaves, E. rossii leaves and E. cupressiformis leaves, and partitioned via sequential extraction with n-hexane, dichloromethane (DCM), ethyl acetate and ethanol. The crude extracts and fractions were screened for antioxidant activity using a novel, high-throughput lipid-based antioxidant assay, as well as the aqueous ABTS (2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assay and the Folin-Ciocalteu test for total phenols. In the lipid-based assay, non-polar n-hexane and DCM fractions showed higher antioxidant activity against the formation of peroxides and thiobarbituric acid reactive substances (TBARS) than the other fractions, whereas the non-polar fractions were not effective in aqueous assays. This illustrates that the high potential of the lipid-soluble n-hexane and DCM fractions as antioxidants would have been missed if only aqueous-based assays were used. In addition, the potent antioxidant compounds were putatively annotated using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-qTOF-MS). Gallic acid, (+)-catechin, (-)-epicatechin and tannins were found in most crude extracts.

Metabolomics approach in identifying biomarkers from pathogenic Ganoderma boninense involved in early interactions with oil palm host

This study attempts to identify key biomarkers that may be involved in the early interactions of oil palm and Ganoderma boninense. To achieve this, the oil palm extract medium (OPEM) was adopted as a model to simulate oil palm-Ganoderma interactions in planta with biomarkers identified using the LC-MS-based metabolomics approach. The metabolic profiles of ethyl acetate crude extracts of OPEM and G. boninense-inoculated OPEM (OPEM + Gb, incubated for 1, 3, 7, 14, 21 and 28 days) were compared using liquid chromatography quadrupole/time-of-flight-mass spectrometry (LC-Q/TOF-MS) coupled with multivariate data analysis using the platform MetaboAnalyst. Results revealed that the OPEM constituents such as kakuol, kanokoside A, abruquinone B and robinetin 3-rutinoside are naturally occurring plant metabolites and may be involved in defense mechanisms against G. boninense. By contrast, aspulvinone H and methylisocitric acid were produced by G. boninense and may be precursors for the synthesis of other compounds that contribute to successful disease infection. Additionally, the metabolomics analyses further ranked 15 important secondary metabolites according to their variable importance in projection (VIP) scores. These compounds comprise one ketone, two fatty acids, two flavonoids and, one isomer and nine unknowns that are suggestive of early plant-pathogen interactions. These 15 compounds were further analysed using a heatmap revealing decreased concentrations of plant-derived metabolites kakuol, kanokoside A and 5,7,4ʹ-trihydroxy-3,6,8,3ʹ,5ʹ-pentamethoxyflavone and increased concentrations of plant-derived fatty acids 9-hydroxy-hexadecan-1,16-dioic acid and 11-hydroperoxy-12,13-epoxy-9-octadecenoic acid in the presence of G. boninense. Changes in the concentrations of these compounds suggest the presence of G. boninense, thereby indicating early infections in the oil palm. These biomarkers may be further developed as a rapid and non-invasive diagnostic for early detection of Ganoderma infection in oil palm.

Magnetic solid-phase extraction of atrazine with ACC@NiCo2O4@Fe3O4 nanocomposite in spice and water samples

ABSTRACT A simple, environmentally friendly, effective and selective magnetic solid-phase extraction method (MSPE) has been developed for the separation and quantitative determination of atrazine from spices and water samples before liquid chromatographic (LC) analysis. Activated carbon cloth@NiCo2O4@Fe3O4 was characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray photoelectron spectroscopy (XPS), X-ray diffraction spectrometry (XRD), Brunauer–Emmett–Teller (BET), and scanning electron microscopy with energy distribution X-Ray detector (SEM-EDX) techniques. The developed MSPE-LC procedure was optimized in terms of pH, adsorbent amount, adsorption time, eluent type and volume, interference parameters and the pre-concentration factor and extraction efficiency at optimum conditions were determined. In the determination of atrazine, it was observed that the activated carbon cloth@NiCo2O4@Fe3O4 nanocomposite could be reused at least 25 times by applying the adsorption-desorption cycle. According to best extraction efficiency of parameters for the extraction and determination of atrazine from spices and water samples were determined as pH 7, adsorbent amount of 0 mg, total adsorption-desorption time of 5 minutes, and enrichment factor of 20. The recovery values of the magnetic solid-phase extraction method in spices and water samples under optimum conditions are between 91% and 94%. Analysis of trace levels of atrazine, limit of detection (LOD) and limit of quantification (LOQ) were determined by HPLC-DAD method with 0.5 ng mL−1 and 1.7 ng mL−1, respectively. Sample applications with high recovery values performed with the MSPE-LC method were confirmed by liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF/MS) for spices and water samples.

Investigation of dichlorodiphenyltrichloroethane (DDT) on xenobiotic enzyme disruption and metabolomic bile acid biosynthesis in DDT-sprayed areas using wild rats.

Dichlorodiphenyltrichloroethane (DDT) is an organochlorine insecticide used worldwide. Several studies have reported the toxic effects of DDT and its metabolites on steroid hormone biosynthesis; however, its environmental effects are not well understood. This study examined wild rats collected in DDT-sprayed areas of South Africa and quantified plasma metabolites using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS). Fold change analysis of the metabolome revealed the effect of DDT on bile acid biosynthesis. Gene expression of the related enzyme in rat liver samples was also quantified. Significant association was found between DDT and gene expression levels related to constitutive androstane receptor mediated enzymes, such as Cyp2b1 in rat livers. However, our results could not fully demonstrate that enzymes related to bile acid biosynthesis were strongly affected by DDT. The correlation between DDT concentration and gene expression involved in steroid hormone synthesis in testis was also evaluated; however, no significant correlation was found. The disturbance of metabolic enzymes occurred in rat liver in the target area. Our results suggest that DDT exposure affects gene expression in wild rats living in DDT-sprayed areas. Therefore, there is a need for DDT toxicity evaluation in mammals living in DDT-sprayed areas. We could not find an effective biomarker that could reflect the mechanism of DDT exposure; however, this approach can provide new insights for future research to evaluate DDT effects in sprayed areas.

Structure Analysis of the Interfering Substance N-methyl-2-phenylpropan-1-amine of Methamphetamine in Wastewater.

To analyze the chemical structure of the interfering substance that affects the result of methamphetamine analysis in wastewater. A combination of GC-MS and liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-QTOF-MS) was used to analyze the mass spectrum characteristics of the interfering substance that affects the result of methamphetamine analysis and to infer its possible structure. Liquid chromatography-triple quadrupole-mass spectrometry (LC-TQ-MS) was used to confirm the control material. Using LC-QTOF-MS in positive electrospray ionization (ESI+) mode, the mass-to-charge ratio (m/z) of quasi-molecular ion in the MS1 mass spectrometry of interfering substance was identical to that of methamphetamine, indicating that the interfering substance was probably an isomer of methamphetamine. The MS2 mass spectra obtained at three collision energies of 15 V, 30 V and 45 V were highly similar to methamphetamine, suggesting that the interfering substance contained methylamino and benzyl groups. Further analysis using GC-MS in electron impact (EI) ionization mode showed that the base peak in the mass spectrum of the interfering substance was at m/z 44. The interfering substance was confirmed to be N-methyl-2-phenylpropan-1-amine by compared with the standard reference. The chemical structure of N-methyl-2-phenylpropan-1-amine is highly similar to methamphetamine, which is easy to cause interference for the detection of trace amounts of methamphetamine in wastewater using LC-TQ-MS. Therefore, in the actual analysis, the chromatographic retention time can be used to distinguish between N-methyl-2-phenylpropan-1-amine and methamphetamine.

Anti-Acetylcholinesterase and Synergistic Antifungal Activities of Selected Salvia Species: Correlation with Metabolic Profiles

Salvia species are known for their anti-fungal and anti-acetylcholinesterase (AChE) activity. Metabolomics is defined as a comprehensive quantitative and qualitative analysis of large-scale of metabolites. In this study, besides determining anti-acetylcholinesterase and synergistic antifungal activities of selected Salvia species (S. cryptantha Montbret & Aucher ex Benth., S. tchihatcheffii (Fisch. & C. A. Mey.) Boiss., S. officinalis L., S. virgata Jacq.), the metabolite profiles were clarified and correlation analyzes between the activity results and profiles were carried out. Aerial parts of all plant materials were extracted by methanol to determine the metabolic profile by using GC-MS (Gas Chromatography-Mass Spectrometry) and LC-QTOF-MS (Liquid Chromatography Quadrupole Time of Flight Mass Spectrometry). Also, Ellman's spectrophotometric method for anti-acetylcholinesterase activity and the checkerboard method for synergistic antifungal activity between extracts and fluconazole were performed. 295 known and 1408 unknown metabolites were detected by using GC-MS while 346 known and 69008 unknown metabolites were by using LC-QTOF-MS. S. cryptantha, S. tchihatcheffii, S. officinalis, and S. virgata demonstrated inhibitory activities on AChE with the ratio of 32.72%, 5.9%, 43.96%, and 12.1% respectively at the concentration of 200 µg/mL. A synergism was found for all Salvia sp. against Candida tropicalis ATCC 750 strain (FICI values of ≤0.5). In addition, primary and secondary metabolites, which are highly correlated with both activities, were determined. All Salvia species exhibited synergic activity with fluconazole while having weak AChE inhibitory activity. Combining natural products with synthetic drugs is important in order to increase efficacy. At the same time, herbal products must be used carefully at the point of herb-drug interactions. Further studies are needed to investigate active metabolites according to correlation results.

Exposure to polylactic acid induces oxidative stress and reduces the ceramide levels in larvae of greater wax moth (Galleria mellonella)

Plastic biodegradation by insects has made significant progress, opening up new avenues for the treatment of plastic waste. Wax moth larvae, for example, have attracted the attention of the scientific community because they are known to chew, ingest, and biodegrade natural polymer bee waxes. Despite this, we know very little about how these insects perform on manufactured plastics or how manufactured plastics affect insect metabolism. As a result, we studied the metabolism of greater wax moths (Galleria mellonella) fed on molasses-supplemented polylactic acid plastic (PLA) blocks. An analysis of the central carbon metabolism (CCM) metabolites was performed using liquid chromatography triple quadrupole mass spectrometry (LC-QQQ-MS), while an analysis of untargeted metabolites and lipids was conducted using liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS). In total, 169 targeted CCM metabolites, 222 untargeted polar metabolites, and 196 untargeted nonpolar lipids were identified within the insect samples. In contrast, compared to control larvae, PLA-fed larvae displayed significantly different levels of 97 CCM metabolites, 75 polar metabolites, and 57 lipids. Purine and pyrimidine metabolisms were affected by PLA feeding, as well as amino acid metabolism, carbohydrates, cofactors, vitamins, and related metabolisms. Additionally, PLA exposure disrupted insect energy metabolism and oxidative stress, among other metabolic disturbances. The larvae fed PLA have lower levels of several lipids, suggesting a reduction in lipid reserves, and ceramide levels are likely to have changed due to apoptosis and inflammation. The study indicates that G. mellonella larvae could ingest PLA but this process causes some metabolic stress for the host. Future studies of the molecular pathways of this biodegradation process might help to provide strategies for stress reduction that would speed up insect digestion of plastic.

Human phase-I metabolism and prevalence of two synthetic cannabinoids bearing an ethyl ester moiety: 5F-EDMB-PICA and EDMB-PINACA.

Around 2017, with the appearance of 5F-EDMB-PINACA, synthetic cannabinoids (SCs) carrying an ethyl ester moiety at the linked group started spreading on the market of new psychoactive substances (NPS). In 2020 and 2021, the indole analog of 5F-EDMB-PINACA (5F-EDMB-PICA) and the non-fluorinated analog of this compound (EDMB-PINACA) were analytically characterized. Here, we present suitable urinary markers to prove the consumption of these two ethyl analogs. Ten authentic urine samples for each compound were analyzed by liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-qToF-MS). Anticipated phase-I metabolites detected in urine samples were confirmed in vitro by applying a pooled human liver microsomes (pHLM) assay. Prevalence data were obtained from urines collected for abstinence control and submitted to a screening method for SC metabolites. Ten phase-I metabolites of 5F-EDMB-PICA and 18 of EDMB-PINACA were detected by LC-qToF-MS analysis of authentic urine specimens. The main in-vivo metabolites were built by ester hydrolysis, often coupled to further metabolic processes. Investigation of phase-I biotransformation led to the identification of ester hydrolysis, monohydroxylation, and defluorination products as the most suitable urinary biomarkers for 5F-EDMB-PICA. Metabolites formed by ester hydrolysis coupled to ketone formation and by monohydroxylation are suggested for the detection of EDMB-PINACA. From October 1, 2020 to February 1, 2022, among positive urine samples, 5.4% and 10.1% tested positive 5F-EDMB-PICA and EDMB-PINACA, respectively. Due to common metabolites shared among structurally related SCs, the unequivocal detection of their consumption remains challenging for forensic laboratories and requires sensitive methods to monitor multiple metabolites, ideally including highly specific species.